OA21436A - Pyrrolopyrimidine amines as complement inhibitors - Google Patents

Abstract

Disclosed are compounds of formula (I), and pharmaceutical acceptable salts thereof, which are inhibitors of the complement system. Also provided are pharmaceutical compositions comprising such a compound, and methods of using the compounds and compositions in the treatment or prevention of a disease or condition characterized by aberrant complement system activity

Description

PYRROLOPYRIMIDINE AMINES AS COMPLEMENT INHIBITORS

RELATED APPLICATION

This application daims the benefit of priority to U.S. Provisional Patent Application serial number 63/004,799, fïled April 3, 2020.

BACKGROUND OF THE INVENTION

The complément System is a branch of an organism’s immune System that enhances the ability of antibodies and phagocytic cells to destroy and remove foreign particles (e.g., 10 pathogens) from the organism. The complément System comprises a set of plasma proteins that act together to attack extracellular forms of pathogens and induce a sériés of inflammatory responses to help fight infection. Complément activation can occur through several pathways. For example, complément activation can occur spontaneously in response to certain pathogens or by antibody binding to a pathogen. When complément proteins are 15 activated a cascade is triggered by which one complément protein induces the activation of the next protein in the sequence. The activation of a small number of complément proteins at the start of the pathway is hugely amplifïed by each successive enzymatic reaction, resulting in the rapid génération of a disproportionately large complément response. (Marrides, S. Pharmacological Reviews 1998, Vol. 50, pages 59-88). In healthy organisme there are 20 regulatory mechanisms to prevent uncontrolled complément activation.

When activated, complément proteins can bind to a pathogen, opsonizing them for engulfment by phagocytes bearing receptors for complément. Then, small fragments of some complément proteins act as chemoattractants to recruit more phagocytes to the site of complément activation, and also to activate these phagocytes. Next, the complément proteins 25 create holes or pores in the invading organisme, leading to their destruction. While complément plays an important rôle in protecting the body from foreign organisms, it can also destroy healthy cells and tissue. The inappropriate activation of complément is implicated in a long list of disease pathologies (Morgan, B. Eur J Clin Invest 1994, Vol. 24, pages 219-228) affecting the immune, rénal, cardiovascular, and neurological Systems.

Accordingly, there exists a need to develop further complément inhibitors, which hâve therapeutic potential in the treatment of numerous disorders.

SUMMARY OF THE INVENTION

In certain aspects, the invention provides compounds having the structure of formula (I), and pharmaceutically acceptable salts thereof:

^r2 (i);

wherein, independently for each occurrence:

X is a bond or C(R^X)2;

Y is a bond, C(R^Y)₂, or -N(R^b)~;

G is S or C(R³)₂;

R^a and R^b are each independently H or (Ci-Cô)alkyl;

R¹ représente optionally substituted aryl, heteroaryl, alkyl, cycloalkyl, alkenyl, or cycloalkenyl;

R² represents optionally substituted bicyclic or tricyclic heteroaryl;

R³ is independently for each occurrence H, halogen, -CN, -NH₂, -CH₂NH₂, (CiCé)alkoxy or (Ci-Côjalkyl; or two vicinal occurrences of R³ taken together with the carbon atoms to which they are bonded form an optionally substituted fused (Ca-Cîjcycloalkyl or (Cé)aryl; or two geminal occurrences of R³ taken together with the carbon atom to which they are bonded form an optionally substituted spiro (C3-C7)cycloalkyl; or two hominal occurrences of R³ taken together with the carbon atoms to which they are bonded form an optionally substituted bridged (C3-C7)cycloalkyl;

R^x is independently for each occurrence H, (Ci-C6)alkyl, or (C3-C7)cycloalkyl;

R^Y is independently for each occurrence H, (Ci-Ce)alkyl, or (C3-C7)cycloalkyl;

or an occurrence of R^Y and a substituent on R² taken together with the intervening atoms form a ring;

optional substituents on R¹ or R² each independently represent halogen, -CN, -NO2,

-OR¹³, ~NR¹³R, -C(O)R¹³, -C(O)OR¹³, -C(O)NR¹³R¹⁴, -OC(O)R¹³, -NR¹³C(O)Rⁱ⁴, -OC(O)NR¹³R¹⁴, -OC(O)OR¹³, -NR¹³C(O)OR¹⁴, -NR¹³C(O)NR¹³R¹⁴, -OS(O)p(R¹³), -SR¹³,-NR¹³S(O)p(R¹⁴), or optionally substituted alkyl, alkenyl, alkynyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cycloalkoxyalkyl, aryloxyalkyl, aralkyl, heteroaralkyl, heteroaryl, aryl, aryloxy, cycloalkyl, (cycloalkyl)alkyl, heterocycloalkyl, or (heterocycloalkyl)alkyl;

or wherein two substituents on R¹ or two substituents on R², taken together with the intervening atoms, form a ring;

R¹³ and R¹⁴, independently for each occurrence, represent H or optionally substituted alkyl, haloalkyl, alkenyl, alkynyl, aryl, or heteroaryl; and p is 0, 1, or 2.

In certain aspects, the invention provides a pharmaceutical composition, comprising a compound of the invention, or a pharmaceutically acceptable sait thereof; and a pharmaceutically acceptable carrier.

In certain aspects, the invention provides methods of treating a disease or condition characterized by aberrant complément System activity, comprising administering to a subject in need thereof a therapeutically effective amount of a compound the invention, or a pharmaceutically acceptable sait thereof. In certain embodiments, the disease or condition characterized by aberrant complément System activity is an immunological disorder. In certain embodiments, the disease or condition characterized by aberrant complément System activity is a disease of the central nervous System. In certain embodiments, the disease or condition characterized by aberrant complément System activity is a neurodegenerative disease or neurological disease. In certain embodiments, the disease or condition characterized by aberrant complément System activity is a rénal disease. In certain embodiments, the disease or condition characterized by aberrant complément System activity is a cardiovascular disease. In certain embodiments, the disease or condition characterized by aberrant complément System activity is a cardiometabolic disease. In certain embodiments, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of paroxysmal noctumal hemoglobinuria, atypical hemolytic urémie syndrome, organ transplant rejection, myasthenia gravis, neuromyelitis optica, membranoproliferative glomerulonephritis, dense-deposit disease, cold agglutinin disease, and catastrophic antiphospholipid syndrome. In certain other aspects, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of adult respiratory distress syndrome, myocardial infarct, lung inflammation, 5 sepsis, cardiopulmonary bypass, bums, asthma, restenosis, multiple organ dysfunction,

Guillain-Barré syndrome, hémorrhagie shock, glomerulonephritis, systemic lupus erythematosus, rheumatoid arthritis, infertility, Alzheimer’s disease, multiple sclerosis, platelet storage, and hemodialysis. In certain other aspects, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of 10 antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), warm autoimmune hemolytic anémia, IgA nephropathy, C3 glomerulonephritis, and focal segmentai glomerulosclerosis. In further aspects, the disease or condition characterized by aberrant complément System activity is a hematological disorder. In further aspects, the disease or condition characterized by aberrant complément System activity is an ocular disorder or an eye disorder. In still further aspects, the disease or condition characterized by aberrant complément System activity is macular degeneration, age-related macular degeneration (AMD), wet AMD, géographie atrophy, macular edema, diabetic macular edema, choroidal neovascularization (CNV), uveitis, Behcet’s uveitis, proliférative diabetic retinopathy, non-proliferative diabetic retinopathy, glaucoma, hypertensive retinopathy, a corneal neovascularization disease, post-comeal transplant rejection, a corneal dystrophie disease, an autoimmune dry eye disease, Stevens-Johnson syndrome, Sjogren’s syndrome, an environmental dry eye disease, Fuchs’ endothélial dystrophy, retinal vein occlusion, or postoperative inflammation. In certain other aspects, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of obesity, insulin résistance, diabètes, dyslipidemia, nephropathy, neuropathy, angioedema, e.g., hereditary angioedema or acquired angioedema, thrombotic microangiopathy, Parkinson’s disease, schizophrenia, periodontitis, Crohn’s disease, C3 glomerulopathy, membranous nephropathy, osteoarthritis, bullous pemphigoid, psoriasis, hidradenitis suppurativa, of ischemia/reperfiision injury, acute kidney injury, and organ transplantation, e.g., kidney transplant, systemic inflammatory response syndrome, septic shock, trauma, cancer, antibody-mediated rejection, Berger’s disease, delayed graft fonction, granulomatosis with polyangiitis, graft versus host disease, hematopoietic stem cell transplant-related thrombotic microangiopathy, immune complex-mediated membranoproliferative glomerulonephritis,

immune-mediated necrotizing myopathy, idiopathic polypoidal choroidal vasculopathy, microscopie polyangitis, pyoderma gangrenosum, and Stargardt Disease 1.

DETAILED DESCRIPTION

Inhibitors of the complément System are useful in therapeutic methods and compositions suitable for use in treating disorders of the immune, rénal, cardiovascular, and neurological Systems. Provided herein are compounds of formula (I) and pharmaceutically acceptable salts thereof that are useful in treating or preventing a disease or condition characterized by aberrant activity of the complément System.

Définitions

The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

The term “heteroatom” is art-recognized and refers to an atom of any element other than carbon or hydrogen. Illustrative heteroatoms include boron, nitrogen, oxygen, phosphorus, sulfur and sélénium, and altematively oxygen, nitrogen or sulfur.

The term “alkyl” as used herein is a term of art and refers to saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, and cycloalkyl substituted alkyl groups. In certain embodiments, a straight-chain or branched-chain alkyl has about 30 or fewer carbon atoms in its backbone (e.g., C1-C30 for straight chain, C3-C30 for branched chain), and altematively, about 20 or fewer, or 10 or fewer. In certain embodiments, the term “alkyl” refers to a C1-C10 alkyl group. In certain embodiments, the term “alkyl” refers to a Ci-Cô alkyl group, for example a Ci-Ce straight-chain alkyl group. In certain embodiments, the term “alkyl” refers to a C3-C12 branched-chain alkyl group. In certain embodiments, the term “alkyl” refers to a C3-C8 branched-chain alkyl group. Représentative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, and n-hexyl.

The term “cycloalkyl” means mono- or bicyclic or bridged saturated carbocyclic rings, each having from 3 to 12 carbon atoms. Certain cycloalkyls hâve from 5-12 carbon atoms in their ring structure, and may hâve 6-10 carbons in the ring structure. Preferably, cycloalkyl is (C3-C7)cycloalkyl, which represents a monocyclic saturated carbocyclic ring, having from 3 to 7 carbon atoms. Examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. Bicyclic cycloalkyl ring Systems include bridged monocyclic rings and fused bicyclic rings. Bridged monocyclic rings contain a monocyclic cycloalkyl ring where two non-adjacent carbon atoms of the monocyclic ring are linked by an alkylene bridge of between one and three additional carbon atoms (Le., a bridging group of the form -(CHijw-, where w is 1, 2, or 3). Représentative examples of bicyclic ring Systems include, but are not limited to, bicyclo[3.1.1]heptane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, bicyclo[3.3.1]nonane, and bicyclo[4.2.1]nonane. Fused bicyclic cycloalkyl ring Systems contain a monocyclic cycloalkyl ring fused to either a phenyl, a monocyclic cycloalkyl, a monocyclic cycloalkenyl, a monocyclic heterocyclyl, or a monocyclic heteroaryl. The bridged or fused bicyclic cycloalkyl is attached to the parent molecular moiety through any carbon atom contained within the monocyclic cycloalkyl ring. Cycloalkyl groups are optionally substituted. In certain embodiments, the fused bicyclic cycloalkyl is a 5 or 6 membered monocyclic cycloalkyl ring fused to either a phenyl ring, a 5 or 6 membered monocyclic cycloalkyl, a 5 or 6 membered monocyclic cycloalkenyl, a 5 or 6 membered monocyclic heterocyclyl, or a 5 or 6 membered monocyclic heteroaryl, wherein the fused bicyclic cycloalkyl is optionally substituted.

The term “(cycloalkyl)alkyl” as used herein refers to an alkyl group substituted with one or more cycloalkyl groups. An example of cycloalkylalkyl is cyclohexylmethyl group.

The term “heterocycloalkyl” as used herein refers to a radical of a non-aromatic ring System, including, but not limited to, monocyclic, bicyclic, and tricyclic rings, which can be completely saturated or which can contain one or more units of unsaturation, for the avoidance of doubt, the degree of unsaturation does not resuit in an aromatic ring System, and having 3 to 12 atoms including at least one heteroatom, such as nitrogen, oxygen, or sulfur. For purposes of exemplification, which should not be construed as limiting the scope of this invention, the following are examples of heterocyclic rings: aziridinyl, azirinyl, oxiranyl, thiiranyl, thiirenyl, dioxiranyl, diazirinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3dithiolanyl, 1,3-dithianyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, azetyl, oxetanyl, oxetyl, thietanyl, thietyl, diazetidinyl, dioxetanyl, dioxetenyl, dithietanyl, dithietyl, dioxalanyl, oxazolyl, thiazolyl, triazinyl, isothiazolyl, isoxazolyl, azepines, azetidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, oxopiperidinyl, oxopyrrolidinyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, quinuclidinyl, thiomorpholinyl, tetrahydropyranyl, tetrahydrofuranyl, tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1-dioxidothiomorphoIinyl (thiomorpholine sulfone), thiopyranyl, trithianyl, and 2azobicyclo[3.1.0]hexane. A heterocycloalkyl group is optionally substituted by one or more substituents as described below.

The term “(heterocycloalkyl)alkyl” as used herein refers to an alkyl group substituted with one or more heterocycloalkyl (i.e., heterocyclyl) groups. _r

The term “alkenyl” as used herein means a straight or branched chain hydrocarbon radical containing from 2 to 10 carbons and containing at least one carbon-carbon double bond formed by the removal of two hydrogens. Représentative examples of alkenyl include, but are not limited to, ethenyl, 2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4-pentenyl, 5hexenyl, 2-heptenyl, 2-methyl-l-heptenyl, and 3-decenyl. The unsaturated bond(s) of the alkenyl group can be located anywhere in the moiety and can hâve either the (Z) or the (E) configuration about the double bond(s).

The term “alkynyl” as used herein means a straight or branched chain hydrocarbon radical containing from 2 to 10 carbon atoms and containing at least one carbon-carbon triple bond. Représentative examples of alkynyl include, but are not limited, to acetylenyl, 1propynyl, 2-propynyl, 3-butynyl, 2-pentynyl, and 1-butynyl.

The term “alkylene” is art-recognized, and as used herein pertains to a diradical obtained by removing two hydrogen atoms of an alkyl group, as defined above. In one embodiment an alkylene refers to a disubstituted alkane, i.e., an alkane substituted at two positions with substituents such as halogen, azide, alkyl, arylalkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldéhyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, fluoroalkyl (such as trifluromethyl), cyano, or the like. That is, in one embodiment, a “substituted alkyl” is an “alkylene”.

The term “amino” is a term of art and as used herein refers to both unsubstituted and substituted amines, e.g., a moiety that may be represented by the general formulas:

R_a

R_a | ₊ --h/ --N—R_b ^Rb and ^Rc , wherein Ra, Rb, and Rc each independently represent a hydrogen, an alkyl, an alkenyl, -(CFhjx-Rd, or Ra and Rb, taken together with the N atom to which they are attached

complété a heterocycle having from 4 to 8 atoms in the ring structure; Ra représente an aryl, a heteroaryl, a cycloalkyl, a cycloalkenyl, a heterocyclyl or a polycyclyl; and x is zéro or an integer in the range of 1 to 8. In certain embodiments, only one of R_a or Rb may be a carbonyl, e.g., Ra, Rb, and the nitrogen together do not form an imide. In other embodiments,

Ra and Rb (and optionally R_c) each independently represent a hydrogen, an alkyl, an alkenyl, or -(CH2)_X-Rd. In certain embodiments, Ra and Rb are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, (cycloalkyl)alkyl, (heterocycloalkyl)alkyl, arylalkyl, heteroarylalkyl, alkoxyalkyl, or haloalkyl, any of which may be further substituted (e.g., by halogen, alkyl, alkoxy, hydroxy, and so forth). In certain embodiments, the term “amino” refers to -NH2.

In certain embodiments, the term “alkylamino” refers to -NH(alkyl).

In certain embodiments, the term “dialkylamino” refers to -N(alkyl)2.

The term “amido”, as used herein, means -NHC(=O)-, wherein the amido group is bound to the parent molecular moiety through the nitrogen. Examples of amido include alkylamido such as CH₃C(=O)N(H)- and CH₃CH₂C(=O)N(H)-.

The term “acyl” is a term of art and as used herein refers to any group or radical of the form RCO- where R is any organic group, e.g., alkyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl. Représentative acyl groups include acetyl, benzoyl, and malonyl.

The term “aminoalkyl” as used herein refers to an alkyl group substituted with one or more one amino groups. In one embodiment, the term “aminoalkyl” refers to an aminomethyl group, i.e., -CH2NH2.

The term “aminoacyl” is a term of art and as used herein refers to an acyl group substituted with one or more amino groups.

The term “aminothionyl” as used herein refers to an analog of an aminoacyl in which 25 the O of RC(O)- has been replaced by sulfur, hence is of the form RC(S)-.

The term “phosphoryl” is a term of art and as used herein may in general be represented by the formula:

Q50 II -P—

OR59 wherein Q50 represents S or O, and R59 represents hydrogen, a lower alkyl or an aryl; for example, -P(O)(OMe)- or -P(O)(OH)2. When used to substitute, e.g., an alkyl, the phosphoryl group of the phosphorylalkyl may be represented by the general formulas:

Q50Q50

--Q51—p--ο--- --Q51—P—OR59 II

OR59OR59 wherein Q50 and R59, each independently, are defined above, and Q51 represents O, S or N; for example, -O-P(O)(OH)OMe or -NH-P(0)(0H)2. When Q50 is S, the phosphoryl moiety is a “phosphorothioate.”

The term “aminophosphoryl” as used herein refers to a phosphoryl group substituted with at least one amino group, as defined herein; for example, -P(0)(0H)NMe2.

The term “azide” or “azido”, as used herein, means an -N3 group.

The term “carbonyl” as used herein refers to -C(=O)-.

The term “thiocarbonyl” as used herein refers to -C(=S)-.

The term “alkylphosphoryl” as used herein refers to a phosphoryl group substituted with at least one alkyl group, as defined herein; for example, -P(O)(OH)Me.

The term “alkylthio” as used herein refers to alkyl-S-. The term “(alkylthio)alkyl” refers to an alkyl group substituted by an alkylthio group.

The term “carboxy”, as used herein, means a -CO2H group.

The term “aryl” is a term of art and as used herein refers to includes monocyclic, bicyclic and polycyclic aromatic hydrocarbon groups, for example, benzene, naphthalene, anthracene, and pyrene. Typically, an aryl group contains from 6-10 carbon ring atoms (i.e., (Cô-Cio)aryl). The aromatic ring may be substituted at one or more ring positions with one or more substituents, such as halogen, azide, alkyl, arylalkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldéhyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, fluoroalkyl (such as trifluromethyl), cyano, or the like. The term “aryl” also includes polycyclic ring Systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings (the rings are “fused rings”) wherein at least one of the rings is an aromatic hydrocarbon, e.g., the other cyclic rings may be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. In certain embodiments, the term “aryl” refers to a phenyl group.

The term “heteroaryl” is a term of art and as used herein refers to a monocyclic, bicyclic, and polycyclic aromatic group having 3 to 12 total atoms including one or more heteroatoms such as nitrogen, oxygen, or sulfur in the ring structure. Exemplary heteroaryl groups include azaindolyl, benzo(b)thienyl, benzimidazolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, benzoxadiazolyl, furanyl, imidazolyl, imidazopyridinyl, indolyl, indohnyl, indazolyl, isomdolmyl, isoxazolyl, isothiazolyl, isoquinolinyl, oxadiazolyl, oxazolyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrrolyl, pyrrolo[2,3-d]pyrimidinyl, pyrazolo[3,4-d]pyrimidinyl, quinolinyl, quinazolinyl, triazolyl, thiazolyl, thiophenyl, tetrahydroindolyl, tetrazolyl, thiadiazolyl, thienyl, thiomorpholinyl, triazolyl or tropanyl, and the like. The “heteroaryl” may be substituted at one or more ring positions with one or more substituents such as halogen, azide, alkyl, arylalkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldéhyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, fluoroalkyl (such as trifluromethyl), cyano, or the like. The term “heteroaryl” also includes polycyclic ring Systems having two or more cyclic rings in which two or more atoms are common to two adjoining rings (the rings are “fused rings”) wherein at least one of the rings is an aromatic group having one or more heteroatoms in the ring structure, e.g., the other cyclic rings may be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Such heteroaryl groups may be connected to the rest of the molécule through either the aromatic group having one or more heteroatoms in the ring structure or the other cyclic group. For example, heteroaryl includes indole, which comprises a benzene ring and a pyrrole ring that are fused together. An indole substituent may be attached to a parent structure through either the benzene ring or through the pyrrole ring of the indole.

The term “aralkyl” or “arylalkyl” is a term of art and as used herein refers to an alkyl group substituted with an aryl group, wherein the moiety is appended to the parent molécule through the alkyl group.

The term “heteroaralkyl” or “heteroarylalkyl” is a term of art and as used herein refers to an alkyl group substituted with a heteroaryl group, appended to the parent molecular moiety through the alkyl group.

The term “alkoxy” as used herein means an alkyl group, as defined herein, appended to the parent molecular moiety through an oxygen atom. Représentative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, tert-butoxy, pentyloxy, and hexyloxy.

The term “alkoxyalkyl” refers to an alkyl group substituted by an alkoxy group.

The term “alkoxycarbonyl” means an alkoxy group, as defined herein, appended to the parent molecular moiety through a carbonyl group, represented by -C(=O)-, as defined herein. Représentative examples of alkoxycarbonyl include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, and ZerZ-butoxycarbonyl.

The term “alkylcarbonyl”, as used herein, means an alkyl group, as defined herein, appended to the parent molecular moiety through a carbonyl group, as defined herein. Représentative examples of alkylcarbonyl include, but are not limited to, acetyl, 1-oxopropyl, 2,2-dimethyl-l-oxopropyl, 1-oxobutyl, and 1-oxopentyl.

The term “arylcarbonyl”, as used herein, means an aryl group, as defined herein, appended to the parent molecular moiety through a carbonyl group, as defined herein. Représentative examples of arylcarbonyl include, but are not limited to, benzoyl and (2pyridinyl)carbonyl.

The term “alkylcarbonyloxy” and “arylcarbonyloxy”, as used herein, means an alkylcarbonyl or arylcarbonyl group, as defined herein, appended to the parent molecular moiety through an oxygen atom. Représentative examples of alkylcarbonyloxy include, but are not limited to, acetyloxy, ethylcarbonyloxy, and ZerZ-butylcarbonyloxy. Représentative examples of arylcarbonyloxy include, but are not limited to phenylcarbonyloxy.

The term “alkenoxy” or “alkenoxyl” means an alkenyl group, as defined herein, appended to the parent molecular moiety through an oxygen atom. Représentative examples of alkenoxyl include, but are not limited to, 2-propen-l-oxyl (i.e., CH2=CH-CH2-Ô-) and vinyloxy (i.e., CH2=CH-0-).

The term “aryloxy” as used herein means an aryl group, as defined herein, appended to the parent molecular moiety through an oxygen atom.

The term “heteroaryloxy” as used herein means a heteroaryl group, as defined herein, appended to the parent molecular moiety through an oxygen atom.

The term “carbocyclyl” as used herein means a monocyclic or multicyclic (e.g., bicyclic, tricyclic, etc.) hydrocarbon radical containing from 3 to 12 carbon atoms that is completely saturated or has one or more unsaturated bonds, and for the avoidance of doubt, the degree of unsaturation does not resuit in an aromatic ring System (e.g., phenyl). Examples of carbocyclyl groups include 1-cyclopropyl, 1-cyclobutyl, 2-cyclopentyl, 1cyclopentenyl, 3-cyclohexyI, 1-cyclohexenyl and 2-cyclopentenylmethyl.

The term “cyano” is a term of art and as used herein refers to -CN.

The term “halo” is a term of art and as used herein refers to -F, -Cl, -Br, or -I.

The term “haloalkyl” as used herein refers to an alkyl group, as defined herein, wherein some or ail of the hydrogens are replaced with halogen atoms.

The term “hydroxy” is a term of art and as used herein refers to -OH.

The term “hydroxyalkyl”, as used herein, means at least one hydroxy group, as defined herein, is appended to the parent molecular moiety through an alkyl group, as defïned herein. Représentative examples of hydroxyalkyl include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, 2,3-dihydroxypentyl, and 2-ethyl-45 hydroxyheptyl.

The term “silyl”, as used herein, includes hydrocarbyl dérivatives of the silyl (H3S1-) group (i.e., (hydrocarbyl)aSi-), wherein a hydrocarbyl groups are univalent groups formed by removing a hydrogen atom from a hydrocarbon, e.g., ethyl, phenyl. The hydrocarbyl groups can be combinations of differing groups which can be varied in order to provide a number of 10 silyl groups, such as trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tertbutyldimethylsilyl (TBS/TBDMS), triisopropylsilyl (TIPS), and [2(trimethylsilyl)ethoxy]methyl (SEM).

The term “silyloxy”, as used herein, means a silyl group, as defined herein, is appended to the parent molécule through an oxygen atom.

Certain compounds contained in compositions of the présent invention may exist in particular géométrie or stereoisomeric forms. In addition, compounds of the présent invention may also be optically active. The présent invention contemplâtes ail such compounds, including cis- and Zra«5-isomers, (7?)- and (S)-enantiomers, diastereoisomers, (D)-isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, as falling 20 within the scope of the invention. Additional asymmetric carbon atoms may be présent in a substituent such as an alkyl group. Ail such isomers, as well as mixtures thereof, are intended to be included in this invention.

If, for instance, a particular enantiomer of compound of the présent invention is desired, it may be prepared by asymmetric synthesis, or by dérivation with a chiral auxiliary, 25 where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers. Altematively, where the molécule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographie 30 means well known in the art, and subséquent recovery of the pure enantiomers.

It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, fragmentation, décomposition, cyclization, élimination, or other reaction.

The term “substituted” is also contemplated to include ail permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described herein above. The permissible substituents may be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may hâve hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. This invention is not intended to be limited in any manner by the permissible substituents of organic compounds.

In certain embodiments, the optional substituents contemplated in this invention include halogen, azide, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, (cycloalkyl)alkyl, heterocyclyl, (heterocyclyl)alkyl, hydroxyl, alkoxyl, amino, aminoalkyl, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether (e.g., -alkylene-O(alkyl)), alkylthio, sulfonyl, sulfonamido, ketone (e.g., CO(alkyl)), aldéhyde (-C(O)H), ester (e.g., -COO(alkyl)), haloalkyl, hydroxyalkyl, alkoxyalkyl, haloalkoxy, haloalkoxyalkyl, and cyano.

As used herein, the term “optionally substituted” or “substituted or unsubstituted” when it précédés a list of Chemical moieties means that the list of Chemical moieities that follow are each substituted or unsubstituted. For example, “substituted or unsubstituted aryl, heteroaryl, and cycloalkyl” or “optionally substituted aryl, heteroaryl, and cycloalkyl” means substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted cycloalkyl.

The term “vicinal” describes the positional relationship. between two substituents, moieties, or functional groups each of which is bonded to one of two adjacent carbon atoms that are bonded to each other (i.e., the two substituents, moieties, or functional groups are in a 1,2-relationship). The methyl groups in 3,4-dimethylheptane are vicinal.

The term “geminal” describes the positional relationship between two substituents, moieties, or functional groups bonded to the same carbon atom (i.e., the two substituents, moieties, or functional groups are in a 1,1-relationship). The methyl groups in 3,3dimethylheptane are geminal.

The term “hominal” describes the positional relationship between two substituents, moieties, or functional groups each of which is bonded to one of two carbon atoms that themselves are each bonded to a single carbon atom (i.e., the two substituents, moieties, or functional groups are in a 1,3-relationship). The methyl groups in 3,5-dimethylheptane are 5 hominal.

The phrase “protecting group”, as used herein, means temporary substituents which protect a potentially reactive functional group from undesired Chemical transformations. Examples of such protecting groups include esters of carboxylic acids, silyl ethers of alcohols, and acetals and ketals of aldéhydes and ketones, respectively. The fïeld of protecting group chemistry has been reviewed (Greene, T.W.; Wuts, P.G.M. Protective Groups in Organic Synthesis, 2^nd ed.; Wiley: New York, 1991). Protected forms of the inventive compounds are included within the scope of this invention.

For purposes of the invention, the Chemical éléments are identified in accordance with the Periodic Table of the Eléments, CAS version, Handbook of Chemistry and Physics, 67th 15 Ed., 1986-87, inside cover.

Other chemistry terms herein are used according to conventional usage in the art, as exemplified by The McGraw-Hill Dictionary of Chemical Terms (ed. Parker, S., 1985), McGraw-Hill, San Francisco, incorporated herein by reference). Unless otherwise defined, ail technical and scientific terms used herein hâve the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

The term “pharmaceutically acceptable sait” as used herein includes salts derived from inorganic or organic acids including, for example, hydrochloric, hydrobromic, sulfuric, nitric, perchloric, phosphoric, formic, acetic, lactic, maleic, fumaric, succinic, tartaric, glycolic, salicylic, citric, methanesulfonic, benzenesulfonic, benzoic, malonic, trifluoroacetic, 25 trichloroacetic, naphthalene-2-sulfonic, and other acids. Pharmaceutically acceptable sait forms can include forms wherein the ratio of molécules comprising the sait is not 1:1. For example, the sait may comprise more than one inorganic or organic acid molécule per molécule of base, such as two hydrochloric acid molécules per molécule of compound of Formula I. As another example, the sait may comprise less than one inorganic or organic acid molécule per molécule of base, such as two molécules of compound of Formula I per molécule of tartaric acid.

The term “prodrug” as used herein refers to a compound that can be metabolized in vivo to provide a compound of formula I. Thus prodrugs include compounds that can be prepared by modifying one or more functional groups in a compound of formula I to provide a corresponding compound that can be metabolized in vivo to provide a compound of formula I. Such modifications are known in the art. For example, one or more hydroxyl groups or amine groups in a compound of formula I can be acylated with alkyl-C(=O)groups or with residues from amino acids to provide a prodrug.

Prodrug forms of a compound bearing various nitrogen-containing fiinctional groups (amino, hydroxyamino, amide, etc.) may include the following types of dérivatives, where each R_p group individually may be hydrogen, substituted or unsubstituted alkyl, aryl, alkenyl, alkynyl, heterocycle, alkylaryl, arylalkyl, aralkenyl, aralkynyl, cycloalkyl or cycloalkenyl.

(a) Carboxamides, represented as —NHC(O)R_P (b) Carbamates, represented as —NHC(O)OR_P (c) (Acyloxy)alkyl Carbamates, represented as NHC(O)OROC(O)R_P (d) Enamines, represented as —NHCR(=CHCO₂R_P) or —NHCR(=CHCONR_pRp) (e) Schiff Bases, represented as —N=CR_pRp (f) Mannich Bases (from carboximide compounds), represented as RCONHCH₂NR_PR_P

Préparations of such prodrug dérivatives are discussed in various literature sources (examples are: Alexander et al., J. Med. Chem. 1988, 31,318; Aligas-Martin et al., PCT W00041531, p. 30).

Prodrug forms of carboxyl-bearing compounds include esters (—CO₂Rm), where the R_m group corresponds to any alcohol whose release in the body through enzymatic or hydrolytic processes would be at pharmaceutically acceptable levels. Another prodrug derived from a carboxylic acid form of the disclosure may be a quaternary sait type of structure described by Bodor et aL, J. Med. Chem. 1980, 23, 469.

The terms “carrier” and “pharmaceutically acceptable carrier” as used herein refer to a diluent, adjuvant, excipient, or vehicle with which a compound is administered or formulated for administration. Non-limiting examples of such pharmaceutically acceptable carriers include liquids, such as water, saline, and oils; and solids, such as gum acacia, gelatin, starch paste, talc, keratin, colloïdal silica, urea, and the like. In addition, auxiliary, stabilizing, thickening, lubricating, flavoring, and coloring agents may be used. Other examples of suitable pharmaceutical carriers are described in Remington ’s Pharmaceutical Sciences by E.W. Martin, herein incorporated by reference in its entirety.

The term “treat” as used herein means prevent, hait or slow the progression of, or eliminate a disease or condition in a subject. In one embodiment “treat” means hait or slow the progression of, or eliminate a disease or condition in a subject. In one embodiment,

“treat” means reduce at least one objective manifestation of a disease or condition in a subject.

The term “effective amount” as used herein refers to an amount that is sufficient to bring about a desired biological effect.

The term “therapeutically effective amount” as used herein refers to an amount that is sufficient to bring about a desired therapeutic effect.

The term “inhibif ’ as used herein means decrease by an objectively measurable amount or extent. In various embodiments “inhibif ’ means decrease by at least 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 95 percent compared to relevant control. In one embodiment “inhibit” means decrease 100 percent, i.e., hait or eliminate.

The term “subject” as used herein refers to a mammal. In various embodiments, a subject is a mouse, rat, rabbit, cat, dog, pig, sheep, horse, cow, or non-human primate. In one embodiment, a subject is a human.

Compounds

The présent invention provides compounds having the structure of Formula (I), and pharmaceutically acceptable salts thereof:

wherein, independently for each occurrence:

X is a bond or C(R^X)2; .

Y is a bond, C(R^Y)2, or -N(R^b)-;

G is S or C(R³)₂;

R^a and R^b are each independently H or (Ci-Côjalkyl;

R¹ représente optionally substituted aryl, heteroaryl, alkyl, cycloalkyl, alkenyl, or cycloalkenyl;

R² représente optionally eubstituted bicyclic or tricyclic heteroaryl;

R³ is independently for each occurrence H, halogen, -CN, -NH2, -CH2NH2, (CiCô)alkoxy or (Ci-Côjalkyl; or two vicinal occurrences of R³ taken together with the carbon atoms to which they are bonded form an optionally substituted fused (C3-C7)cycloalkyl or (Cô)aryl; or two geminal occurrences of R³ taken together with the carbon atom to which they are bonded form an optionally substituted spiro (Cs-Cîjcycloalkyl; or two hominal occurrences of R³ taken together with the carbon atoms to which they are bonded form an optionally substituted bridged (C3-C7)cycloalkyl;

R^x is independently for each occurrence H, (Ci-Ce)alkyl, or (C3-C7)cycloalkyl;

R^Y is independently for each occurrence H, (Ci-Ce)alkyl, or (C3-C7)cycloalkyl;

or an occurrence of R^Y and a substituent on R² taken together with the intervening atoms form a ring;

optional substituents on R¹ or R² each independently represent halogen, -CN, -NO2, -OR¹³, -NR¹³R¹⁴, -C(O)R¹³, -C(O)OR¹³, -C(O)NR¹³R¹⁴, -OC(O)R¹³, -NR¹³C(O)R¹⁴, -OC(O)NR¹³R¹⁴, -OC(O)OR¹³, -NR^I3C(O)OR¹⁴, -NR¹³C(O)NR¹³R¹⁴, -OS(O)p(R¹³), -SR¹³,-NR^I3S(O)p(R¹⁴), or optionally substituted alkyl, alkenyl, alkynyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cycloalkoxyalkyl, aryloxyalkyl, aralkyl, heteroaralkyl, heteroaryl, aryl, aryloxy, cycloalkyl, (cycloalkyl)alkyl, heterocycloalkyl, or (heterocycloalkyl)alkyl;

or wherein two substituents on R¹ or two substituents on R², taken together with the intervening atoms, form a ring;

R¹³ and R¹⁴, independently for each occurrence, represent H or optionally substituted alkyl, haloalkyl, alkenyl, alkynyl, aryl, or heteroaryl; and p is 0, 1, or 2.

In certain embodiments, Y represents C(R^Y)2, preferably CH2.

In certain embodiments, X represents a bond. Altematively, X may represent CH2.

In certain embodiments, R¹ represents optionally substituted aryl or heteroaryl. In some embodiments, R¹ is optionally substituted heteroaryl. Exemplary embodiments of R¹ include optionally substituted phenyl (e.g., 3-halophenyl, or 2,3-dihalophenyl), (C321436

Cejcycloalkyl, alkenyl, pyridinyl (e.g., 6-halopyridin-2-yl), or pyrazinyl (e.g., 6-halopyrazin2-yl).

In certain embodiments, R¹ is mono-, di-, or tri- substituted.

In certain embodiments, R¹ représente optionally substituted pyridinyl, preferably optionally substituted 2-pyridinyl. For example, R¹ may represent

In other embodiments, R¹ represents optionally substituted phenyl. For example, R¹

Z¹ represents N or CR^1Z;

Z² represents N or CR^2Z;

Z³ represents N or C;

Z⁴ represents N or CR^4Z;

Z⁵ represents N or CR^5Z;

Z⁶ represents N or CR^6Z;

Z⁷ represents N or CR^7Z;

Z⁸ représente C;

Z⁹ represents N or C;

k is an integer from 1-4;

m is an integer from 1-3; and each occurrence of R^1Z, R^2Z, R^4Z, R^5Z, R^6Z, R^7Z, R^2A independently represents H, halogen, -CN, -NO2, -OR¹³, -NR¹³R¹⁴, -C(O)R¹³, -C(O)OR¹³, -C(O)NR¹³R¹⁴, -OC(O)R¹³, -NR^I3C(O)R¹⁴, -OC(O)NR¹³R¹⁴, -OC(O)OR¹³, -NR¹³C(O)OR¹⁴, -NR¹³C(O)NR¹³Rⁱ⁴, -OS(O)p(R¹³), -SR¹³, -NR¹³S(O)_P(R¹⁴), or optionally substituted alkyl, alkenyl, alkynyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cycloalkoxyalkyl, aryloxyalkyl, aralkyl, heteroaralkyl, heteroaryl, aryl, aryloxy, cycloalkyl, (cycloalkyl)alkyl, heterocycloalkyl, or (heterocycloalkyl)alkyl; or wherein an occurrence of R^6Z and an occurrence of R^7Z taken together with the intervening atoms form a ring; or wherein an occurrence of R^Y and an occurrence of R^2Z taken together with the intervening atoms form a ring.

In certain embodiments, Z², Z³, Z⁴, and Z⁶ represent N.

In certain embodiments, Z³, Z⁴, and Z⁶ represent N.

In certain embodiments, Z⁹, Z⁴, and Z⁶ represent N.

In certain embodiments, Z³ and Z⁴ represent N.

In certain embodiments, Z¹, Z³, Z⁴, and Z⁶ represent N.

r^5Z

In certain embodiments, R2 represents In certain embodiments, R2 represents In certain embodiments, R2 represents CD nh2 5 In certain embodiments, R2 represents In certain embodiments, R2 represents nOO nh2

IXYR“ r7Z N=/ R\/N ¥ Yï/) ΐζ Riz. r5<Y^¥ I ¥ /W r7Z , for example R5Z .N. J Y Ï Yr“ r7Z R1Z \ H z^R2Z । 77 R1Z R , for example

In certain embodiments, R^7Z represents -NR¹³R¹⁴, for example -NH2.

In certain embodiments, R^6Z représente -C(O)R¹³, -C(O)OR¹³, -C(O)NR¹³R¹⁴, or hydroxy alkyl.

In certain embodiments, R^5Z represents alkyl, halo, or -NR¹³R¹⁴.

In certain embodiments, R^1Z represents -CN, halo, haloalkyl, optionally substituted 5 aryl, optionally substituted heteroaryl, optionally substituted alkyl, -C(O)R¹³, -SR¹³, -NR¹³R¹⁴, -OR¹³, -C(O)OR¹³, -C(O)NR¹³R¹⁴, or-NR¹³C(O)R¹⁴.

In certain embodiments, each occurrence of R^2A independently represents -CN, -NO2, halo, haloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl, -C(O)R¹³, -C(O)OR¹³, -NR¹³C(O)OR¹⁴, -SR¹³, -NR¹³R¹⁴, -OR¹³, -C(O)NR¹³R¹⁴, or -NR¹³C(O)R¹⁴.

In certain embodiments, G is C(R³)2.

In certain embodiments, the compound of the invention has the structure of formula (la):

R² (la).

In certain embodiments, two vicinal occurrences of R³ taken together with the carbon atoms to which they are bonded form an optionally substituted fused Cs-cycloalkyl.

In certain embodiments, at least one occurrence of R³ is halo, preferably fluoro.

In certain embodiments, at least one occurrence of R³ is methyl.

In certain embodiments, two vicinal occurrences of R³ taken together with the carbon atoms to which they are bonded form an optionally substituted fused Ca-cycloalkyl and another occurrence of R³ is methyl. .

In certain embodiments, the compound of formula (I) is selected from the following table of compounds, and pharmaceutically acceptable salts thereof:

#

Structure

#

Structure

81a	X-^N0 F nh2	35f	i\r° νύΜ f XTS N==Z X^7 nh2
82a	V 1 H \A^N N Br r o LJ 0 oc? nhch3	30f	m Z J tzz. w° <C /Ρ~ΎΧ 1 z“\ te/ Ί x
83d	JW N Cy^N\ h2n	47g	w 0 1 cooch, νύΜ nV nh2
84a	<>» ¥| H N-WN^-N^Br / T Ύ Ί X o U. Λ ÇQ nh2	48d	w nWv^ / Λ NH2 Byj
85b	-'“'Z'-, Y 1 H \WNVN ^Br J il II Ί !\ O H. .X ° ço nh2	38e	Ζ^?Λ I \ / t>J / ( x? ><3 œ 0=^ ZI Γ z V-4 00

138d	O kjf xz y=o /° |^JZ— \ ( CN VA X z y—z \=z	181a	Vr i? Τ'/T\— Z5^/ λ—z 1 M ° j o=\ ZI ά ro -1
139b	0 ° F il Ί /> nh2 N^NH	182a	'Wu* ^'v-N LQv y 'L N NH2 '—/
125d	QyliOÔ /-A o nh2 f	183a	τ \\ .-''Λ Μ Z y \ o=\ ZI / Z 00
126f	Z—ç Z Z \_/ hî } f w y-7 o=\	207b	E.' / N-< H / V—N o yNyBr \-s7 h2n cf3
142d	ÇXyN^jÇXc| O=\ O F H3C N N Z h η n nh2	208d	Wo‘ Ν=Ζν-^ <\ yz> Ν-γ ' CF, NH2 3

146a	,Z=\ Z—(\ Z °yjÇz 2 ° p ω y o=\ ZI \ 71 -
147a	I ω O O o=/ Z—ά Z I // Nï )-----f 0^0’ ο=λ &
148d	F ,Ν. Ν Λ O ° F Îl J /) NH2 1
19a	F N-4 H /Γ\-νΤ) 0 ^V-Br \==/ H2N VssZ
3b	r? Z—Z \ w Ù ο^ζ^γ,,,,-π oy 00 n

186a	pV rSr N-/U <ζ5-Λ7 N==/ nh2
206c	N—i H J \-N f V-N ο ο 1Γ y-Br h2n cf3
209b	/ N—\ H __/ V-N N / Il \—N /Γ VN x° 0 JT VBr NysZy-, '\==^ h 2N cf3
210d	/ \-N N V V-N /Γ vN o 0 F V-Br h 2n cf3
189g	w Γ o I^Rvie n-vnv< <z TT? nv v=\ nh2 Me

llf	L. DÛ iA z Z iz \=o -L·2 v ΧΓ =c''‘ ^Z )-----( c^ '/Ve 1 z y—z \=Z
12a	f. p o=\ ZI ! -z. V-A en n
127d	J·'·.__ \ 1 H N-y-N N Br rN_NA 0 kj WCO O
128a	W rNvN 0
154e	ffl y~\ z j IZ v=o z° A z— ty vzV” K_£ zv=hz

245a	Λ-Γζ N y \=A nh2
42a	Aro’ i 0 n-VNV^ ¢, __H \yCO2Me n=a \===z nh2
16e	WA l0 i xrycF3 N-W \W nh2
36e	r^0 y /( y-°Me nh2
39f	m AAs z / ! CM iz o yo O wC t/ Ί £ cr

37a	r^0 Ta__ΖΓ Nx^-OMe NV \A nh2	221c	’Wt/ ^n\^n ςχχ NH2 1 f H3COOC
218a	ΖΛ Z \_/ m y ( 8^^ I o^=\ ΖΞ ! z. A 00 —1	250b	1—\ H N'if A A 0 r'0 Az fVP) CN N A nh2
223a	A' 0 1 cooch3 νύΜ f M? N==/ \=7 nh2	270a	// z 0 Æy— z r? Ct CD “I
224b	Br N=/ hn-4 # )£< y—/ '-<N 0 ^=0 n=x Tf N-\ CO2H nh2	271c	____ 11 .. Br °A f x^n n a NH2 rVcH3

274g	N—ί H o=^ # y-N > ° FyBr y-n \^/ N<yJyY-, h2n ocf3	275c	N) —. z-y' V \ Z /Ύ ° A -.'V °Yyy ο V==/ °^f ZI 03
276g	N—x H __/ \—N N / x\ \-—N // vn ο ο [Γ y-Br h2n 0	284a	Wï/ /^° [ί>Ν'ΐΓΝ\—yMe n<=k^vX nh2 n==^
277a	-7 1 NJ ZO 1 ^-/ 1 'O-O'ù / ΟΓ'-ζ'χΙ OeyY ZI 00 ~1	285g	m Va z J IZ Φ \=o s ckQ v Î?
278g	Z zAy t? Y'z—X X^o0 *P ~O \ o=x ZI 1 Z νΛ 00 —1	286a	Z—(/ Z I \_/ M > ( ^A/S Æ// T VzZ // œ U / O=X ZI t Z tu “1
279a	m Y\ z J IZ \=o \ r-T/ΎΎ n?7 x rvH Z\^-Z	287a	CJ z Ί xz \^o f Φ m _Uz A I Y =<'' ^z ί f CM / \ x z />—z

280g	W Λ°ο- <\ Ι-A Ν-γ \=\ nh2 f	288c	/Ά __Me nh2 X==f N
281a	^^Z zâj . >? Jlz^\ XZX-æ P~ -Π \ o=\ ZI / z CD n	289a	tv-©A.“ N-Zv f xrs N λ nh2 y%
282a	w A.° n-<nvA fin n=/ \=\ nh2 oh	290g	m z J xz \=O CD cki?
283g	w rSte Au <'X_A N =γ N^7 nh2	291a	z—C z z \—/ Μ >----( oXJs><3 ω φ 1 O=\ ZI / Z χ-Λ Q

325c	F, Wv/ N-γ \==\ nh2 CF3	326c	1 0 N N^ZvY < M/ N-γ N=\ NH2 CF3
327c	ήλκ/ l0 < T-O Ν-γ N=\ nh2 CF3	328f	r^° n^nv^ / î try° nh2
329a	w A n^/nv^ / <x mY° N ~~27 nh2	330f	Ζ^Ν-Λ km Z—X Z511/ λ— Z Ί A-Y ° j 'ο'χ o^=\ -n ZI / Z VA □3 —1
331a	r^° Ν~γ \==\ NH2 °	336c	r? Z—. z·-// Λ \ Z /Ύ ° Λ—x\ Z JL —·/ zz . ÜD —1
337a	ήχ©·' 0<l f ΙΑ N λ X nh2 Va F	315a	O Y z Y et <T __1 z—v Λ=ϊ~/ \Υ Ί £ r^x^z Vz

316a	W-t?·' 1 O [Hle f X-O N A nh2 N=X Me	343a	’“Wa [Xle νύΜ f LH N=Z \=4 NH2 H N=X Me
344c	W [ 4le 0Me Xu f M? N=/ \=\ nh2 CF3	345c	u. O Z J XZ LL y0 ® z^/° <( /ύΎ 1 z—v X/ Z | .n =< \ U X cr
346g	r^s-<f rV n-yM f V-P.N nV NH2 CF3	347a	A.° </Νχχ$ NH2 CF3
338e	n ,N-Vu 0=K E Y-N > ° frr y-n \^/ H2N XiZ	339g	m x) XZ /X^° 7=\ / =Xz.^^ £ /[ O /===( z \ A //—-z. V-Z £
348c	N-AtA’ f X-O N=/ \=\ NH2 CF3	349c	CO O Va z J XZ V=o Γ ® « X /P Lz -y I T X·''- >--( CM 'X i z />— Z

350a	'b-AY· T 0 N /^° __Me 1 N nh2 x—( cf3	351a	ήν-ο° rV fX^rS1 N =/ \==\ NH2 CF3
332c	r0 <\ λΠ N~~f NH2 CF3	333c	ί X %_ Æ- Y—Z G \ o==^ /zi o τι Z ω Χλ ' ?
365c	’K H μ yCI r^° <x In n-4/ \===ς NH2 CF3	366c	x° n^NV( y<7 N-γ \=\ NH2 CF3
352c	m z J iz >° φ n-o rX ° VxJU K-''' ^z )-----ί cm \ X Z /)— Z	353a	Z—(' Z X \_/ ΙΌ )-----\ •''''ΐ jT ζ·/^]-— °-^ Φ oX ZI / Z VA CO —I

354c	[<Ny-N Me N Tv5 NH2 \ Vcf3	355a	z—(z z z \_/ M )---( z^li o z. ® μ o=\ / ZI o / τι ^=\ ω X / Z 00
340a	r? Z~~. zy Λ /Ύ ° Λ Ji -.·>/ // y z °\ zi 00 Ί	341c	N—i H o=( # y-n > ° fy-Br y-N \^/ h2n L-z vX. L-z^ochs
342a	,ί·\ // Z O /CLZ^se^. y— Z r? Tjz-/ jy. O “ O X ω	356c	w /^° ii>N'VN\—/Me NyWX nh2 v— N cf3
357a	_/?y Z—V z I \_/ M >----f nW O 0=4 J1 ZI l -z. CD 1	358c	Ly /^° __,Me Νν^ν~Λ nh2 cf3 //y V— N

359c	ZI / Z CD	360c	z—(z z z \_/ N> )------< φφ ZI / z CD “1
361a	w ^ΝγΝ Me nh2	367g	i^° Ν~φ \=φ nh2 CF3
368a	1_ CQ ζχχ Z / IZ V y=° xo r i co φ /P Xz Λ £ T )-----( CM Z^X—Z \=Z	369g	WM Af <\ IA Ν~φ nh2 CF3
362f	w r\e N-Λτί fX-O nh2 CF3	363a	Wm Γ^Ίνΐβ wM f X-O N=Z NM nh2 01=3
364e	Xsw I^Rvie ΝύΜ N==/ \==\ NH2 CF3	364d	XX- Î^Xle nwNVÂ < M7 NM \=Φ NH2 CF3

381g	nX H o=( # y-N .n > ° Frr F /X Ν<ΧΧΧ. h2n F	382a	N-\ H o=^ XN\—N .N > ° ΓχΒΓ ^===7 NXXX h2n χχχ F
385a	L. CQ x) XZ \ LL. _/=-o X. XX X π ^ z y-' o /===( z \ A <zz '“——••p CM	384g	N-X H °X VNyN K .n > ° X y-Br ÇX HZN XX~f
386g	CO χ) XZ /-Xo $Xz___X Z Ύ-— O /===( Z \ \\ '/—z z X	387a	X -, M ----, Vk \ z /Y ° // L / * °v T! \ ZX ro —1
383g	k. CÛ x) XZ \ \ O Xz^^ X r11· -V'·^ >[ Z Ny_J/ ° XX Z \ Λ //—z. ^Z £	388a	X z z-~y Λ \ Z /V ° JX -.·»ν // O \ \ ZX œ n
389g	N—x H 0=/ .n > ° Xv N<XXX h2n xXf Br	390a	1? Z-—. 'Z·-// Λ \ Z /Y ° // y z \L· / o^Z7 Tl \ ZX CO “1

Pharmaceutical Compositions

The invention provides pharmaceutical compositions, each comprising one or more compounds of the invention, or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition comprises a compound of the invention, or a pharmaceutically acceptable sait thereof, and a pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition comprises a plurality of compounds of the invention, which may include pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier.

In certain embodiments, a pharmaceutical composition of the invention further comprises at least one additional pharmaceutically active agent other than a compound of the invention. The at least one additional pharmaceutically active agent can be an agent useful in the treatment of a disease or condition characterized by aberrant complément System activity.

Pharmaceutical compositions of the invention can be prepared by combining one or more compounds of the invention with a pharmaceutically acceptable carrier and, optionally, one or more additional pharmaceutically active agents.

Methods of Use

The présent invention provides compounds, and pharmaceutically acceptable salts thereof, that are useful for treating or preventing a disease or condition characterized by aberrant complément System activity.

In certain aspects, the invention provides a compound of the invention, or a pharmaceutically acceptable sait thereof, for use as a médicament.

In certain aspects, the invention provides methods of treating or preventing a disease or condition characterized by aberrant complément System activity. The method includes the step of administering to a subject in need thereof a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable sait thereof, thereby treating or preventing the disease or condition characterized by aberrant complément System activity. By reducing complément System activity in the subject, the disease or condition characterized by aberrant complément System activity is treated.

Altematively, in certain aspects, the invention provides a compound of the invention, 75 or a pharmaceutically acceptable sait thereof, for treatment of a disease or condition characterized by aberrant complément System activity.

Altematively, in certain aspects, the invention provides the use of a compound of the invention, or a pharmaceutically acceptable sait thereof, for the manufacture of a médicament for use in treatment of a disease or condition characterized by aberrant complément System 20 activity.

As used herein, a “disease or condition characterized by aberrant complément System activity” refers to any disease or condition in which it is désirable to reduce complément System activity. For example, it may be désirable to reduce complément System activity in the setting of inappropriate activation or hyperactivation of the complément System.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is an immunological disorder.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is a disease of the central nervous System.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is a neurodegenerative disease or neurological disease.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is a rénal disease.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is a cardiovascular disease.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is a cardiometabolic disease.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of paroxysmal noctumal hemoglobinuria, atypical hemolytic urémie syndrome, organ transplant rejection, myasthenia gravis, neuromyelitis optica, membranoproliferative glomerulonephritis, dense-deposit disease, cold agglutinin disease, and catastrophic antiphospholipid syndrome.

In certain embodiments, the disease or condition is paroxysmal noctumal hemoglobinuria.

In certain embodiments, the disease or condition is atypical hemolytic urémie syndrome.

In certain embodiments, the disease or condition is organ transplant rejection.

In certain embodiments, the disease or condition is myasthenia gravis.

In certain embodiments, the disease or condition is neuromyelitis optica.

In certain embodiments, the disease or condition is membranoproliferative glomerulonephritis.

In certain embodiments, the disease or condition is dense-deposit disease.

In certain embodiments, the disease or condition is cold agglutinin disease.

In certain embodiments, the disease or condition is catastrophic antiphospholipid syndrome.

In other embodiments, the disease or condition characterized by aberrant complément System activity is adult respiratory distress syndrome, myocardial infarct, lung inflammation, 25 hyperacute rejection (transplantation rejection), sepsis, cardiopulmonary bypass, bums, asthma, restenosis, multiple organ dysfunction syndrome, Guillain-Barré syndrome, hémorrhagie shock, paroxysmal noctumal hemoglobinuria, glomerulonephritis, systemic lupus erythematosus, rheumatoid arthritis, infertility, Alzheimer’s disease, organ rejection (transplantation), myasthenia gravis, multiple sclerosis, platelet storage, or hemodialysis.

In other embodiments, the disease or condition characterized by aberrant complément

System activity is selected from the group consisting of antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), warm autoimmune hemolytic anémia, IgA nephropathy, C3 glomerulonephritis, and focal segmentai glomerulosclerosis.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is a hematological disorder.

In other embodiments, the disease or condition characterized by aberrant complément System activity is an ocular disorder or an eye disorder.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is macular degeneration, age-related macular degeneration (AMD), wet AMD, géographie atrophy, macular edema, diabetic macular edema, choroidal neovascularization (CNV), uveitis, Behcet’s uveitis, proliférative diabetic retinopathy, nonproliferative diabetic retinopathy, glaucoma, hypertensive retinopathy, a comeal neovascularization disease, post-comeal transplant rejection, a comeal dystrophie disease, an autoimmune dry eye disease, Stevens-Johnson syndrome, Sjogren’s syndrome, an environmental dry eye disease, Fuchs’ endothélial dystrophy, retinal vein occlusion, or postoperative inflammation.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is angioedema, e.g., hereditary angioedema or acquired angioedema.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is obesity, insulin résistance, diabètes, dyslipidemia, nephropathy, or neuropathy.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), paroxysmal noctumal hemoglobinuria, and thrombotic microangiopathy.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of Alzheimer’s disease, multiple sclerosis, neuromyelitis optica, generalized myasthenia gravis, Guillain-Barré syndrome, Parkinson’s disease, and schizophrenia.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is periodontitis.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is Crohn’s disease.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of asthma, chronic obstructive pulmonary disease, and acute respiratory distress syndrome.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is atherosclerosis.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of age-related macular degeneration (AMD), uveitis, glaucoma, and wet AMD.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is myocardial infarction.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of atypical hemolytic urémie syndrome, C3 glomerulopathy, lupus nephritis, IgA nephropathy, and membranous nephropathy.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of rheumatoid arthritis, osteoarthritis, bullous pemphigoid, psoriasis, hidradenitis suppurativa, and burns.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is hemodialysis.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of ischemia/reperfusion injury, acute kidney injury, and organ transplantation, e.g., kidney transplant.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of systemic inflammatory response syndrome, sepsis, septic shock, trauma, systemic lupus erythematosus, hereditary angioedema, and cancer.

In certain embodiments, the disease or condition characterized by aberrant complément System activity is selected from the group consisting of antibody-mediated rejection, antiphospholipid syndrome, Berger’s disease, C3 glomerulonephritis, cold agglutinin disease, cardiopulmonary bypass, dense-deposit disease, delayed graft function, géographie atrophy, granulomatosis with polyangiitis, graft versus host disease, hematopoietic stem cell transplant-related thrombotic microangiopathy, immune complexée mediated membranoproliferative glomerulonephritis, immune-mediated necrotizing myopathy, idiopathic polypoidal choroidal vasculopathy, microscopie polyangitis, pyoderma gangrenosum, Stargardt Disease 1, and warm type autoimmune hemolytic anémia.

Formulations, Routes of Administration, andDosing

The compounds of the invention, and pharmaceutically acceptable salts thereof, can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient, in a variety of forms adapted to the chosen route of administration, e.g., 5 orally or parenterally, by intravenous, intraperitoneal, intramuscular, topical, or subcutaneous routes. Additional routes of administration are also contemplated by the invention.

Thus, the présent compounds may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, may be compressed into tablets, or may be incorporated directly with the food of the patient's diet. For oral therapeutic administration, the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, élixirs, suspensions, syrups, wafers, and the like. Such compositions and préparations should contain at least 0.1% of active compound. The percentage of the compositions and préparations may, 15 of course, be varied and may conveniently be between about 2% to about 60% of the weight of a given unit dosage form. The amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained.

The tablets, troches, pills, capsules, and the like may also contain the following diluents and carriers: binders such as gum tragacanth, acacia, corn starch or gelatin;

.20 excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnésium stéarate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be présent as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like. A syrup or élixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compound may be incorporated into sustained-release préparations and devices.

The active compound may also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compound or its salts can be prepared in water or physiologically acceptable aqueous solution, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these préparations contain a preservative to prevent the growth of microorganisms.

The pharmaceutical dosage forms suitable for injection or infusion can include stérile aqueous solutions or dispersions or stérile powders comprising the active ingrédient which are adapted for the extemporaneous préparation of stérile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In ail cases, the ultimate dosage form should be stérile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, éthanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants. The prévention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phénol, sorbic acid, thimerosal, and the like. In many cases, it will be préférable to include isotonie agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

Stérile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingrédients enumerated above, as required, followed by filter sterilization. In the case of stérile powders for the préparation of stérile injectable solutions, methods of préparation can include vacuum drying and the freeze drying techniques, which yield a powder of the active ingrédient plus any additional desired ingrédient présent in the previously sterile-filtered solutions.

For topical administration, the présent compounds may be applied in pure form, i.e., 30 when they are liquids. However, it will generally be désirable to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable carrier, which may be a solid or a liquid.

Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina and the like. Useful liquid carriers include water, alcohols or glycols or water-alcohol/glycol blends, in which the présent compounds can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use. The résultant liquid compositions can be applied from absorbent pads, used to 5 impregnate bandages and other dressings, or sprayed onto the affected area using pump-type or aérosol sprayers. .

Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified minerai materials can also be employed with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like, for application directly 10 to the skin of the user.

Examples of useful dermatological compositions which can be used to deliver the compounds of the invention to the skin are known in the art; for example, see Jacquet et al. (U.S. Pat. No. 4,608,392; incorporated herein by reference), Geria (U.S. Pat. No. 4,992,478; incorporated herein by reference), Smith et al. (U.S. Pat. No. 4,559,157; incorporated herein by reference), and Wortzman (U.S. Pat. No. 4,820,508; incorporated herein by reference).

Useful dosages of the compounds of the invention can be determined, at least initially, by comparing their in vitro activity and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animais, to humans are known in the art; for example, see U.S. Pat. No. 4,938,949 (incorporated herein by reference).

The amount of the compound, or pharmaceutically acceptable sait thereof, required for use in treatment will vary not only with the particular compound or sait selected but also with the route of administration, the nature of the condition being treated, and the âge and condition of the patient and will be ultimately at the discrétion of the attendant physician or clinician.

In general, however, a suitable dose will be in the range of from about 0.5 to about

100 mg/kg body weight of the récipient per day, e.g., from about 3 to about 90 mg/kg of body weight per day, from about 6 to about 75 mg per kilogram of body weight per day, from about of 10 to about 60 mg/kg of body weight per day, or from about 15 to about 50 mg/kg of body weight per day.

Compounds of the invention, or pharmaceutically acceptable salts thereof, can be conveniently formulated in unit dosage form; for example, containing 5 to 1000 mg, 10 to 750 mg, or 50 to 500 mg of active ingrédient per unit dosage form. In one embodiment, the invention provides a composition comprising a compound of the invention, or pharmaceutically acceptable salts thereof, formulated in such a unit dosage form. The desired dose may conveniently be presented in a single dose or as divided doses to be admimstered at appropnate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrète loosely spaced administrations.

Compounds of the invention, or pharmaceutically acceptable salts thereof, can also be administered in combination with other therapeutic agents, for example, other agents that are useful for treating or preventing ischemia, blood loss, or reperfusion injury. In certain embodiments, compounds of the invention, and pharmaceutically acceptable salts thereof, can also be administered in combination with one or more other therapeutic agents that are useful for treating or preventing an ocular disorder or eye disorder.

Other delivery Systems can include time-release, delayed release, or sustained release delivery Systems such as are well-known in the art. Such Systems can avoid repeated administrations of the active compound, increasing convenience to the subject and the physician. Many types of release delivery Systems are available and known to those of ordinary skill in the art. Use of a long-term sustained release implant may be désirable. Long-term release, as used herein, means that the delivery System or is implant constructed and arranged to deliver therapeutic levels of the active ingrédient for at least 30 days, and preferably 60 days.

In certain embodiments, a compound of the invention is formulated for intraocular administration, for example direct injection or insertion within or in association with an intraocular medical device. In certain embodiments, a compound of the invention is formulated as an ophthalmic solution. In certain embodiments, a compound of the invention can be administered via ocular delivery, for example, by local ocular administration, including topical, intravitreal, periocular, transscleral, retrobulbar, juxtascleral, suprachoroidal, or sub-tenon administration. A compound of the invention can be administered via ocular delivery either alone or in combination with one or more additional therapeutic agents.

The compounds of the invention may be formulated for depositing into a medical device, which may include any of a variety of conventional grafts, stents, including stent grafts, cathéters, balloons, baskets, or other device that can be deployed or permanently implanted within a body lumen. As a particular example, it would be désirable to hâve devices and methods which can deliver compounds of the invention to the région of a body which has been treated by interventional technique.

In exemplary embodiment, a compound of the invention may be deposited within a medical device, such as a stent, and delivered to the treatment site for treatment of a portion of the body.

Stents hâve been used as delivery vehicles for therapeutic agents (i.e., drugs).

Intravascular stents are generally permanently implanted in coronary or peripheral vessels. Stent designs include those of U.S. Pat. No. 4,733,655 (Palmaz), U.S. Pat. No. 4,800,882 (Gianturco), or U.S. Pat. No. 4,886,062 (Wiktor). Such designs include both métal and polymeric stents, as well as self-expanding and balloon-expandable stents. Stents may also be used to deliver a drug at the site of contact with the vasculature, as disclosed in U.S. Pat.

No. 5,102,417 (Palmaz), U.S. Pat. No. 5,419,760 (Narciso, Jr.), U.S. Pat. No. 5,429,634 (Narciso, Jr.), and in International Patent Application Nos. WO 91/12779 (Medtronic, Inc.) and WO 90/13332 (Cedars-Sanai Medical Center), for example.

The term “deposited” means that the compound is coated, adsorbed, placed, or otherwise incorporated into the device by methods known in the art. For example, the compound may be embedded and released from within (“matrix type”) or surrounded by and released through (“réservoir type”) polymer materials that coat or span the medical device. In the latter example, the compound may be entrapped within the polymer materials or coupled to the polymer materials using one or more the techniques for generating such materials known in the art. In other formulations, the compound may be linked to the surface 20 of the medical device without the need for a coating, for example by means of détachable bonds, and release with time or can be removed by active mechanical or Chemical processes. In other formulations, the compound may be in a permanently immobilized form that présents the compound at the implantation site.

In certain embodiments, the compound may be incorporated with polymer compositions during the formation of biocompatible coatings for medical devices, such as stents. The coatings produced from these components are typically homogeneous and are useful for coating a number of devices designed for implantation.

The polymer may be either a biostable or a bioabsorbable polymer depending on the desired rate of release or the desired degree of polymer stability, but frequently a bioabsorbable polymer is preferred for this embodiment since, unlike a biostable polymer, it will not be présent long after implantation to cause any adverse, chronic local response. Bioabsorbable polymers that could be used include, but are not limited to, poly(L-lactic acid), polycaprolactone, polyglycolide (PGA), poly(lactide-co-glycolide) (PLLA/PGA), poly(hydroxybutyrate), poly(hydroxybutyrate-co-valerate), polydioxanone, polyorthoester, polyanhydride, poly(glycolic acid), poIy(D-lactic acid), poly(L-Iactic acid), poly(D, L-lactic acid), poly(D, L-lactide) (PLA), poly (L-lactide) (PLLA), poly(glycolic acid-co-trimethylene carbonate) (PGA/PTMC), polyethylene oxide (PEO), polydioxanone (PDS), polyphosphoester, polyphosphoester urethane, poly(amino acids), cyanoacrylates, poly(trimethylene carbonate), poly(iminocarbonate), copoly(ether-esters) (e.g., PEO/PLA), polyalkylene oxalates, polyphosphazenes and biomolecules such as fïbrin, fïbrinogen, cellulose, starch, collagen and hyaluronic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, cross linked or amphipathic block copolymers of hydrogels, and other suitable bioabsorbable poplymers known in the art. Also, biostable polymers with a relatively low chronic tissue response such as polyuréthanes, silicones, and polyesters could be used, and other polymers could also be used if they can be dissolved and cured or polymerized on the medical device such as polyolefms, polyisobutylene and ethylene-alphaolefin copolymers; acrylic polymers and copolymers, vinyl halide polymers and copolymers, such as polyvinyl chloride; polyvinylpyrrolidone; polyvinyl ethers, such as polyvinyl methyl ether; polyvinylidene halides, such as polyvinylidene fluoride and polyvinylidene chloride; polyacrylonitrile, polyvinyl ketones; polyvinyl aromatics, such as polystyrène, polyvinyl esters, such as polyvinyl acetate; copolymers of vinyl monomers with each other and olefins, such as ethylene-methyl méthacrylate copolymers, acrylonitrile-styrene copolymers, ABS resins, and ethylene-vinyl acetate copolymers; pyran copolymer; polyhydroxy-propyl-methacrylamidephenol; polyhydroxyethyl-aspartamide-phenol; polyethyleneoxide-polylysine substituted with palmitoyl residues; polyamides, such as Nylon 66 and polycaprolactam; alkyd resins, polycarbonates; polyoxymethylenes; polyimides; polyethers; epoxy resins, polyuréthanes; rayon; rayon-triacetate; cellulose, cellulose acetate, cellulose butyrate; cellulose acetate butyrate; cellophane; cellulose nitrate; cellulose propionate; cellulose ethers; and carboxymethyl cellulose.

Polymers and semipermeable polymer matrices may be formed into shaped articles, such as valves, stents, tubing, prostheses and the like.

In certain embodiments of the invention, the compound of the invention is coupled to a polymer or semipermeable polymer matrix that is formed as a stent or stent-graft device.

Typically, polymers are applied to the surface of an implantable device by spin coating, dipping, or spraying. Additional methods known in the art can also be utilized for this purpose. Methods of spraying include traditional methods as well as microdeposition techniques with an inkjet type of dispenser. Additionally, a polymer can be deposited on an implantable device using photo-patteming to place the polymer on only spécifie portions of the device. This coating of the device provides a uniform layer around the device which allows for improved diffusion of various analytes through the device coating.

In certain embodiments of the invention, the compound is formulated for release from the polymer coating into the environment in which the medical device is placed. Preferably, the compound is released in a controlled manner over an extended time frame (e.g., months) using at least one of several well-known techniques involving polymer carriers or layers to control elution. Some of these techniques are described in U.S. Patent Application 2004/0243225Al, the entire disclosure of which is incorporated herein in its entirety.

Moreover, as described for example in U.S. Pat. No. 6,770,729, which is incorporated herein in its entirety, the reagents and reaction conditions of the polymer compositions can be manipulated so that the release of the compound from the polymer coating can be controlled. For example, the diffusion coefficient of the one or more polymer coatings can be modulated to control the release of the compound from the polymer coating. In a variation on this theme, the diffusion coefficient of the one or more polymer coatings can be controlled to modulate the ability of an analyte that is présent in the environment in which the medical device is placed (e.g., an analyte that facilitâtes the breakdown or hydrolysis of some portion of the polymer) to access one or more components within the polymer composition (and for example, thereby modulate the release of the compound from the polymer coating). Yet another embodiment of the invention includes a device having a plurality of polymer coatings, each having a plurality of diffusion coefficients. In such embodiments of the invention, the release of the compound from the polymer coating can be modulated by the plurality of polymer coatings.

In yet another embodiment of the invention, the release of the compound from the . polymer coating is controlled by modulating one or more of the properties of the polymer composition, such as the presence of one or more endogenous or exogenous compounds, or altematively, the pH of the polymer composition. For example, certain polymer compositions can be designed to release a compound in response to a decrease in the pH of the polymer composition.

Kits

The invention also provides a kit, comprising a compound of the invention, or a pharmaceutically acceptable sait thereof, at least one other therapeutic agent, packaging material, and instructions for administering the compound of the invention or the pharmaceutically acceptable sait thereof and the other therapeutic agent or agents to a mammal to treat or prevent a disease or condition characterized by aberrant complément activity. In one embodiment, the mammal is a human.

It will be understood by one of ordinary skill in the relevant arts that other suitable modifications and adaptations to the compositions and methods described herein are readily apparent from the description of the invention contained herein in view of information known to the ordinarily skilled artisan, and may be made without departing from the scope of the invention or any embodiment thereof.

EXAMPLES

Having now described the présent invention in detail, the same will be more clearly understood by reference to the following examples, which are included herewith for purposes of illustration only and are not intended to be limiting of the invention.

Scheme 1

Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (le)

Step-1: Préparation of iert-butyl 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetate (1b)

A mixture of 9H-pyrimido[4,5-b]indol-4-amine (la) (0.50 g, 2.71 mmol; CAS# 400754-6420 5), ter/-butyl 2-bromoacetate (0.582 g, 2.99 mmol) and CS2CO3 (1.061 g, 3.26 mmol) in DMF (20 mL) was stirred at RT for 3 h, diluted with H2O (30 mL) and extracted with EtOAc (30 mL x 3). The combined organics were washed with H2O (30 mL x 4), brine (30 mL), dried, filtered and concentrated under vacuum and purified by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] to provide tert-butyl 2-(4-amino-9Hpyrimido[4,5-b]indol-9-yl)acetate (1b) (0.64 g, 79 % yield) as a yellow solid; *H NMR (300 MHz, DMSO-î/₆) δ 8.35 (d, J= 7.8 Hz, 1H), 8.29 (s, 1H), 7.55 (d, J= 8.1 Hz, 1H), 7.41 (dd, J = 8.3, 7.1 Hz, 1H), 7.35-7.18 (m, 3H), 5.12 (s, 2H), 1.41 (s, 9H).

Step-2: Préparation of 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (le)

A mixture of iert-butyl 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetate (1b) (0.63 g, 2.112 mmol) and 2,2,2-trifluoroacetic acid (12.12 mL, 31.7 mmol) in DCM (20 mL) was stirred at RT for 16 h and concentrated in vacuum to dryness to afford 2-(4-amino-9H-pyrimido[4,510 b]indol-9-yl)acetic acid (le) (0.74 g, 99%) TFA sait as an orange slid; ’H NMR (300 MHz, DMSO-dô) δ 8.55 (s, 1H), 8.49 (d, J= 7.8 Hz, 1H), 8.43 - 8.20 (m, 2H), 7.77 (d, J= 8.2 Hz, 1H), 7.58 - 7.48 (m, 1H), 7.42 (t, J= 7.6 Hz, 1H), 5.26 (s, 2H); ^I9F NMR (282 MHz, DMSOde) δ -74.71.

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(615 bromo-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (le)

To a solution of 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (le) (50.7 mg, 0.142 mmol) TFA sait, (lR,3S,5R)-N-(6-bromo-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane3-carboxamide (Id) HCl sait [(43 mg, 0.129 mmol); prepared according to the procedure reported by Wiles, Jason A. étal., PCT Int. Appl. (2017), WO 2017035353 Al 20170302;

incorporated by reference] and HATU (59.0 mg, 0.155 mmol) in DMF (5 mL) was added dropwise DIPEA (84 mg, 0.646 mmol) and stirred at RT for 16 h. The reaction mixture was diluted with HzO (30 mL) and extracted with EtOAc (20 mL x 2). The combined organics were washed with 0.5 M NaOH (25 mL), H2O (25 mL x 3), brine (25 mL), dried, filtered and concentrated in vacuum to dryness. The residue obtained was purified by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0-5%] to provide product as a pale oil, which was dissolved in MeOH (5 mL) and purified by reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] to provide (lR,3S,5R)-2-(2-(4-amino-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (le) (48 mg, 71 % yield) HCl sait as a white solid. Ή NMR (300 MHz, DMSO-îZ₆) δ 10.28 (s, 1H,

D2O exchangeable), 8.83 - 8.59 (m, 3H, 2H D2O exchangeable), 8.54 (d, J = 7.9 Hz, 1H), 7.71 (d, J = 8.2 Hz, 1H), 7.62 (d, J = 7.9 Hz, 1H), 7.56 (t, J = 7.8 Hz, 1H), 7.45 (m, 2H), 5.75 (d, J = 17.5 Hz, 1H), 5.47 (d, J = 17.3 Hz, 1H), 4.39 (dd, J = 9.1, 5.1 Hz, 1H), 3.93 - 3.86 (m,

1H), 2.41 (m, 1H), 2.27 (m, 1H), 1.99 (s, 4H), 1.11 (m, 1H), 0.82 (m, 1H); MS (ES+): 520.0 (M+l); (ES-): 518.0 (M-l); Analysis calculated for C₂₄H₂₂BrN7O₂.l.lHC1.2.5H₂O: C, 47.60; H, 4.68; Cl, 6.44; N, 16.19; Found: C, 47.57; H, 4.60; Cl, 6.33; N, 16.19.

Scheme 2

Préparation of ( 1 R,3 S,5R)-2-(2-(4-aminopyrrolo [2,1 -f] [ 1,2,4]triazin-7-yl)acetyl)-N-(6bromo-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (2d)

Step-1: Préparation of ZerZ-butyl 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetate (2b)

To a suspension of zinc (1.842 g, 28.2 mmol) (Note: Zinc dust was activated using aqueous

0.1 M HCl, followed by washing thoroughly with H₂O, EtOH and Et₂O, and drying at 100 °C under vacuum before use) and TMSC1 (0.179 mL, 1.408 mmol) in THF (5 mL) at 50 °C was added dropwise ZerZ-butyl 2-bromoacetate (2.75 g, 14.08 mmol) in THF (5 mL) under an argon atmosphère. The mixture was heated at 50 °C for 30 min cooled to RT and was added via syringe to a suspension of 7-bromopyrrolo[l,2-f][l,2,4]triazin-4-amine (2a) (1.00 g, 4.69 mmol; CAS # 937046-98-5), Pd₂(dba)₃ (0.215 g, 0.235 mmol) and XPhos (0.224 g, 0.469 mmol) in THF (10 mL) at RT under an argon atmosphère and heated at 60 °C under argon for 16 h. The reaction mixture was cooled to RT diluted with EtOAc (30 mL), added solid NH4CI (2 g) stirred at RT for 15-min and filtered to remove insoluble solids. The insoluble residue was washed with EtOAc (20 mL) and the fïltrate was washed with saturated NH4CI (30 mL),

H₂O (30 mL), brine (30 mL), dried, filtered and concentrated in vacuum. The obtained residue was purified by flash column chromatography [SiO₂ gel (24 g), eluting with EtOAc in hexane from 0-50%] to provide ZerZ-butyl 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetate (2b) (0.65 g, 56 % yield) as a yellow solid; Ή NMR (300 MHz, DMSO-îL) δ 7.82 (s, 1H), 7.66 (s, 2H), 6.83 (d, J = 4.3 Hz, 1H), 6.52 (d, J = 4.3 Hz, 1H), 3.84 (s, 2H), 1.40 (s, 9H); MS (ES+): 249 (M+l), (ES-): 247 (M-l).

Step-2: Préparation of 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetic acid (2c)

Compound 2c was prepared according to the procedure reported in step-4 of scheme-1, from tert-butyl 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetate (2b) (0.65 g, 2.62 mmol) using TFA (4.01 mL, 52 mmol) in DCM (20 mL) and stirring at RT for 16 h. The reaction mixture was concentrated in vacuum to afford 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetic acid (2c) (1.05 g) TFA sait as an orange solid; ’H NMR (300 MHz, DMSO-ifc) δ 9.67 (dd, J = 5.7,

2.9 Hz, 1H), 9.11 (s, 1H), 8.18 (s, 1H), 7.37 (d, J = 4.6 Hz, 1H), 6.80 (d, J = 4.5 Hz, 1 H), 3.96 (s, 2H). ¹⁹F NMR (282 MHz, DMSO-Î&) δ -74.71; MS (ES+): 193 (M+l), (ES-): 191 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetyl)-N(6-bromo-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (2d)

Compound 2d was prepared according to the procedure reported in step-3 of scheme-1 from

TFA sait of 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetic acid (2c) (54.7 mg, 0.179 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromo-3-methylpyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (Id) (54 mg, 0.162 mmol), HATU (74.1 mg, 0.195 mmol), DIPEA (105 mg, 0.812 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g) eluting with MeOH in

DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (lR,3S,5R)-2-(2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetyl)-N-(6-bromo-3methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (2d) (20 mg, 26 % yield) as a white solid; Ή NMR (300 MHz, DMSO-Jô) δ 10.25 (s, 1H, D₂O exchangeable), 10.01 (s,

1H, D₂O exchangeable), 9.21 (s, 1H, D₂O exchangeable), 8.20 (s, 1H), 7.64 (d, J = 8.0 Hz,

1H), 7.52 (d, J = 4.6 Hz, 1H), 7.45 (d, J = 7.9 Hz, 1H), 6.84 (d, J = 4.6 Hz, 1H), 4.38 (dd, J = 9.1,5.2 Hz, 1H), 4.31 (d, J =17.0 Hz, 1H), 4.17 (d, J = 17.0 Hz, 1H), 3.68 - 3.66 (m, 1H), 2.36 (m, 1H), 2.24 (m, 1H), 2.07 (s, 3H), 1.92 - 1.77 (m, 1H), 0.96 (m, 1H), 0.62 (m, 1H); MS (ES+): 470.0 (M+l), (ES-): 468.0 (M-l).

Scheme 3

Préparation of (2S,4R)-l-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-3methylpyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (3b)

Compound 3b was prepared according to the procedure reported in step-3 of scheme-1, from

TFA sait of 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (le) (150 mg, 0.421 mmol) in DMF (5 mL) using HCl sait of (2S,4R)-N-(6-bromo-3-methylpyridin-2-yl)-4fluoropyrrolidine-2-carboxamide (3a) [(143 mg, 0.421 mmol); prepared according to the procedure reported by Wiles, Jason A. et al., PCT Int. Appl. (2017), WO 2017035355 Al 20170302; incorporated by reference], HATU (192 mg, 0.505 mmol) DIPEA (272 mg, 2.105 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse-phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (2S,4R)-l-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromo-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (3b) (123 mg, 56 % yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-dô) δ (a mixture of two rotamers) 10.92 (s) and 10.48 (s) (2s, 1H, D2O exchangeable), 8.73 (s, 3H, D2O exchangeable), 8.62 (s, 1H),

8.53 (d, J= 7.8 Hz, 1H), 7.70 (d, J= 8.0 Hz, 1H), 7.67 - 7.49 (m, 2H), 7.43 (t, J= 7.9 Hz, 2H), 5.67 (d, J= 4.0 Hz, 1H), 5.63 - 5.45 (m, 1H), 5.37 (d, J= 17.3 Hz, 1H), 4.57 (t, J= 9.4, 7.6 Hz, 1H), 4.48 - 4.23 (m, 1H), 4.20 - 3.95 (m, 1H), 2.79 - 2.49 (m, 4H), 2.32 - 2.05 (m, 1H); ^I9F NMR (282 MHz, DMSO-îZ₆) δ -176.05, -176.37; MS (ES+): 526.0 (M+l), 524.0 (M20 1); Analysis calculated for C23H2iBrFN₇O₂.HCl 2.75H₂O: C, 45.11; H, 4.53; Cl, 5.79; N,

16.01; Found: C, 45.12; H, 4.29; Cl, 5.71; N, 15.84.

Scheme 4

Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4b)

Compound 4b was prepared according to the procedure reported in step-3 of scheme-1, from

TFA sait of 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (le) (150 mg, 0.421 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) [(134 mg, 0.421 mmol); prepared according to the procedure reported by Altmann, Eva et al., PCT Int. Appl. (2012), WO 2012093101 Al 20120712; incorporated by reference], HATU (192 mg, 0.505 mmol), DIPEA (272 mg, 2.105 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (lR,3S,5R)-2-(2-(4-amino-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-315 carboxamide (4b) (110 mg, 52 % yield) HCl sait as a white solid. *H NMR (300 MHz, DMSO-î/ô) δ 10.77 (s, 1H, D₂O exchangeable), 8.63 (m, 3H, 2H D₂O exchangeable), 8.53 (d, J = 7.9 Hz, 1H), 8.01 (d, J = 8.1 Hz, 1H), 7.76 - 7.66 (m, 2H), 7.58 (t, J = 7.5 Hz, 1H), 7.44 (t, J = 7.5 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.78 (d, J = 17.4 Hz, 1H), 5.44 (d, J = 17.3 Hz, 1H), 4.43 (dd, J = 9.0, 5.5 Hz, 1H), 3.93 (m, 1H), 2.44 - 2.16 (m, 2H), 2.01 - 1.86 (m, 1H), 1.08 (m, 1H), 0.78 (m, 1H); MS (ES+) 506.0 (M+l); (ES-): 504.0 (M-l); Analysis calculated for:

C₂3H₂oBrN₇0₂.HCl. 2.25H₂O: C, 47.36; H, 4.41; Cl, 6.08; N, 16.81; Found: C, 47.25; H, 4.18; Cl, 6.02; N, 16.70.

Scheme 5

Préparation of (2S,4R)-1 -(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(6bromopyridin-2-yl)-4-fIuoropyrrolidine-2-carboxamide (5e)

Step-3: Préparation of ZerZ-butyl 2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (5b)

Compound 5b was prepared according to the procedure reported in step-1 of scheme-1, from 7H-pyrrolo[2,3-d]pyrimidin-4-amine (5a) (1.0 g, 7.45 mmol; CAS # 1500-85-2) in DMF (20 mL) using ZerZ-butyl 2-bromoacetate (1.600 g, 8.20 mmol), CS2CO3 (2.91 g, 8.95 mmol) and stirring at RT for 3 h. This gave after workup and purification by flash column chromatography. [silica (24 g), eluting with 0-3% MeOH in DCM] ZerZ-butyl 2-(4-amino-7H10 pyrrolo[2,3-d]pyrimidin-7-yl)acetate (5b) (0.62 g, 34 % yield) as a yellow solid; *H NMR (300 MHz, DMSO-cfc) δ 8.04 (s, 1H), 7.12 (d, J = 3.5 Hz, 1H), 6.99 (s, 2H), 6.54 (d, J = 3.5 Hz, 1H), 4.86 (s, 2H), 1.41 (s, 9H); MS (ES+): 249 (M+l).

Step-4: Préparation of 2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (5c)

Compound 5c was prepared according to the procedure reported in step-2 of scheme-1, from 15 ZerZ-butyl 2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (5b) (0.62 g, 2.497 mmol) using TFA (19.11 mL, 49.9 mmol) in DCM (20 mL) and stirring at RT for 16 h. This gave after workup 2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (5c) (1.42 g) TFA sait as an orange solid; ’H NMR (300 MHz, DMSO-J₆) δ 9.00 (s, 2H), 8.39 (s, 1H), 7.50 (d, J = 3.5 Hz, 1H), 6.93 (d, J= 3.5 Hz, 1H), 5.05 (s, 2H); MS (ES+): 193 (M+l); (ES-): 191 (M-l).

Step-5: Préparation of (2S,4R)-l-(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(6bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (5e)

Compound 5e was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (5c) (137 mg, 0.448 . mmol) in DMF (5 mL) using TFA sait of (2S,4R)-N-(6-bromopyridin-2-yl)-4fluoropyrrolidine-2-carboxamide (5d) [(150 mg, 0.373 mmol); prepared according to the procedure reported by Wiles, Jason A. et al., PCT Int. Appl. (2017), WO 2017035348 Al 20170302; incorporated by reference], HATU (170 mg, 0.448 mmol), DIPEA (241 mg, 1.865 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with 0 - 5% MeOH in DCM] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (2S,4R)-l-(2-(4-amino-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (5e) (84 mg, 49% yield) as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ (a mixture of two rotamers) 11.36 (s) and 11.03 (s) (2s, 1H, D2O exchangeable), 9.22 (s, 2H, D2O exchangeable), 8.36 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.40 (d, J= 3.5 Hz, 1H), 7.33 (d, J= 7.7 Hz, 1H), 6.93 (d, J= 3.5 Hz, 1H), 5.66 - 5.39 (m, 1H), 5.33 (d, J= 17.1 Hz, 1H), 5.18 (d, J= 17.1 Hz, 1H), 4.65 (t, J= 8.5 Hz, 1H), 4.26-4.06 (m, 1H), 3.89 (ddd, J= 38.2, 12.2, 3.0 Hz, 1H), 2.63 - 2.43 (m, 1H), 2.25 - 1.95 (m, 1H); ¹⁹FNMR(282

MHz, DMSO-îZô) δ -175.70, -175.96; MS (ES+): 462.0 (M+l), 460.0 (M-l); Analysis calculated for Ci8H17BrFN7O2.HCl.2H2O: C, 40.43; H, 4.15; Cl, 6.63; N, 18.33; Found: C, 40.19; H, 3.86; Cl, 6.98; N, 17.96.

Scheme 6

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo [3.1.0]hexane-3 -carboxamide (6e)

Step-1: Préparation of 6-methyl-9H-pyrimido[4,5-b]indol-4-amine (6b)

To a solution of triethyl orthoformate (47.7 mL, 450 mmol), AcOH (6.43 mL, 112 mmol) was added 2-amino-5-methyl-lH-indole-3-carbonitrile (6a) (3.85 g, 22.49 mmol) and NH4OAC (8.67 g, 112 mmol) in a 350-mL pressure vessel and heated at 100 °C under pressure for 16 h. The reaction mixture was diluted with H2O (20 mL) and stirred for 15 min. The mixture was then filtered and the solid was washed H2O and air dried to provide βίο methyl-9H-pyrimido[4,5-b]indol-4-amine (6b) (3.60 g, 81%) as apale-yellow solid; ’H NMR (300 MHz, DMSO-Jô) δ 11.67 (s, 1H), 8.22 (s, 1H), 8.12 (s, 1H), 7.32 (d, J= 8.2 Hz, 1H), 7.17 (dd, J= 8.3, 1.5 Hz, 1H), 7.07 (s, 2H), 2.45 (s, 3H); MS (ES+): 1.99 (M+l), (ES-): 197 (M-l).

Step-2: Préparation of ierZ-butyl 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate 15 (6c)

Compound 6c was prepared according to the procedure reported in step-1 of scheme-1, from 6-methyl-9H-pyrimido[4,5-b]indol-4-amine (6b) (2.00 g, 10.09 mmol; CAS # 1242140-67-5) in DMF (20 mL) using fôrt-butyl 2-bromoacetate (2.362 g, 12.11 mmol), CS2CO3 (6.57 g, 20.18 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (20 g), eluting with MeOH in DCM from 0-3%] teri-butyl 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (6c) (2.57 g, 82% yield) as a pale yellow solid; *H NMR (300 MHz, DMSO-ùfe) δ 8.26 (s, 1H), 8.18 (s, 1H), 7.42 (d, J= 8.3 Hz, 1H), 7.29 - 7.15 (m, 3H), 5.08 (s, 2H), 2.47 (s, 3H), 1.40 (s, 9H); MS (ES+): 313 (M+l), (ES-): 311 (M-l).

Step-3: Préparation of 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (6d)

Compound 6d was prepared according to the procedure reported in step-2 of scheme-1, from fëri-butyl 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (6c) (0.220 g, 0.704 mmol) using TFA (8.03 mg, 7.04) in DCM (10 mL) and stirring at RT for 16 h. This gave after workup 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (6d) (300 mg) TFA sait as a purple solid; *H NMR (300 MHz, DMSO-cfe) δ 8.54 (d, J= 1.1 Hz, 1H), 8.48 10 8.26 (m, 3H), 7.67 (d, J= 8.4 Hz, 1H), 7.38 (d, J= 8.6 Hz, 1H), 5.24 (s, 2H); ¹⁹F NMR (282

MHz, DMSO-ifc) δ -74.24; MS (ES+): 257 (M+l), (ES-): 255 (M-l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridih-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (6e)

Compound 6e was prepared according to the procedure reported in step-3 of scheme-1, from 15 TFA sait of 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (6d) (50 mg, 0.135 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (43 mg, 0.135 mmol), HATU (61.6 mg, 0.162 mmol), DIPEA (87 mg, 0.675 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with 0 - 5% MeOH in

DCM] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (lR,3S,5R)-2-(2-(4amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (6e) (48 mg, 68 % yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/₆) δ 10.76 (s, 1H, D₂O exchangeable), 8.88 (s, 2H, D₂O exchangeable), 8.68 (s, 1H), 8.40 (s, 1H), 8.01 (d, J= 8.1 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.63 (d, J= 8.4 Hz, 1H), 7.42 (dd, J= 8.3, 1.6 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.76 (d, J= 17.3 Hz, 1H), 5.41 (d, J= 17.1 Hz, 1H), 4.42 (dd, J= 9.0, 5.5 Hz, 1H), 3.92 (m, 1H), 2.34 (m, 1H), 2.21 (m, 1H), 1.92 (m, 1H), 1.07 (m, 1H), 0.77 (m, 1H); MS (ES+): 520.0 (M+l); (ES-): 518.0 (M-l); Analysis calculated for C₂₄H₂₂BrN₇O₂.1.25HC1.2.75H₂O: C, 46.83; H,

4.71; Cl, 7.20; N, 15.93; Found: C, 46.68; H, 4.62; Cl, 7.26; N, 15.75.

Scheme 7

Préparation of (2S,4R)-l-(2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(3chloro-2-fIuorobenzyl)-4-fluoropyrroIidine-2-carboxamide (7b)

Compound 7b was prepared according to the procedure reported in step-3 of scheme-1, from

TFA sait of 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (6d) (50 mg, 0.135 mmol) in DMF (5 mL) using TFA sait of (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (7a) [(52.5 mg, 0.135 mmol); prepared according to the procedure reported in Wiles, Jason A. et al., PCT Int. Appl. (2017), WO 2017035349 Al 20170302; incorporated by reference], HATU (61.6 mg, 0.162 mmol) DIPEA (0.118 mL,

0.675 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (2S,4R)-l-(2-(4-amino-6methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-475 fluoropyrrolidine-2-carboxamide (7b) (30 mg, 43 % yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-Jô) δ (a mixture of two rotamers) 9.12 (s) and 8.73 - 8.47 (m)(s and m, 4H, 2H exchangeable), 8.37 (s, 1H), 7.60 (d, J= 8.4 Hz, 1H), 7.46 - 7.35 (m, 1H), 7.35 7.25 (m, 1H), 7.14 (td, J = 8.8, 8.4, 3.0 Hz, 1H), 6.86 (t, J= 7.9 Hz, 1H), 5.65 - 5.49 (m, 1H), 5.46 - 5.37 (m, 1H), 5.32 (d, J= 17.3 Hz, 1H), 4.51 - 4.13 (m, 4H), 4.00 (ddd, J= 37.1, 12.7,

3.1 Hz, 1H), 2.45 (s, 4H), 2.25 - 1.90 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-ifc) δ -121.26, 121.77, -176.26, -176.48; MS (ES+): 513.0 (M+l), 511.0 (M-l); Analysis calculated for C25H23CIF2N6O2.HCI.2H2O: C, 51.29; H, 4.82; Cl, 12.11; N, 14.36; Found: C, 51.22; H, 4.90; Cl, 12.01; N, 14.43.

Scheme 8

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8b)

Compound 8b was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (6d) (50 mg,

0.135 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) [(44.9 mg, 0.135 mmol); prepared according to the procedure reported by Wiles, Jason A. et al., PCT Int. Appl. (2017), WO 2017035353 Al 20170302], HATU (61.6 mg, 0.162 mmol), DIPEA (87 mg, 0.675 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with 0-5% MeOH in DCM] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (lR,3S,5R)-2-(2-(4-amino-6-methyl-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-275 azabicyclo[3.1.0]hexane-3-carboxamide (8b) (40 mg, 55 % yield) HCl sait as a white solid; 'HNMR (300 MHz, DMSO-cfe) δ 10.76 (s, 1H, D₂O exchangeable), 8.57 (s, 1H), 8.47 (s, 2H, D₂O exchangeable), 8.33 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.57 (d, J = 8.4 Hz, 1H), 7.37 (d, J= 8.4 Hz, 1H),7.31 (d,J=7.7Hz, 1H), 5.68 (d, J= 17.3 Hz, 1H), 5.34 (d, J= 17.2 Hz, 1H), 4.36 (dd, J= 9.0, 5.9 Hz, 1H), 3.71 - 3.66 (m, 1H), 2.59 - 2.39 (m,

4H), 1.97 (dd, J= 13.2, 5.8 Hz, 1H), 1.30 (s, 3H), 1.05 - 0.96 (m, 1H), 0.96 - 0.88 (m, 1H);

MS (ES+) 534.0 (M+l), 532.0 (M-l).

Scheme 9

HATU, DIPEA

Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (9a)

Compound 9a was prepared according to the procedure reported in step-3 of scheme-1, from

TFA sait of 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yI)acetic acid (le) (50 mg, 0.14 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (8a) (46.7 mg, 0.14 mmol), HATU (80 mg, 0.211 mmol), DIPEA (0.122 mL, 0.702 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in

DCM from 0-100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (9a) (49 mg, 67% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.78 (s, 1H, D₂O exchangeable), 8.77 (s, 2H,

D₂O exchangeable), 8.66 (s, 1H), 8.54 (d, J- 7.9 Hz, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.75 7.67 (m, 2H), 7.61 - 7.55 (m, 1H), 7.45 (t, J= 7.5 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.75 (d, J= 17.4 Hz, 1H), 5.39 (d, J= 17.3 Hz, 1H), 4.38 (dd, J= 9.1, 5.9 Hz, 1H), 3.71 (dd, J= 5.5, 2.4 Hz, 1H), 2.49-2.41 (m, 1H), 1.99 (dd, J= 13.3, 5.9 Hz, 1H), 1.31 (s, 3H), 1.02 (t,J= 5.4 Hz, 1H), 0.94 (dd, J= 5.4, 2.4 Hz, 1H); MS (ES+): 520.0 (M+l), 542.0 (M+Na); (ES-):

518.0 (M-l); Analysis calculated for C₂₄H₂₂BrN₇O₂.1.75(H₂O).1.25(HCl): C, 48.25; H, 4.51;

Cl, 7.42; N, 16.41; Found: C, 48.06; H, 4.27; Cl, 7.28; N, 16.27.

Scheme 10

SS

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (10b)

Compound 10b was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (6d) (50 mg, 0.135 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6-bromo-3methylpyridin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (10a) [(46.7 mg, 0.14 mmol); prepared according to the procedure reported by Wiles, Jason A. et al., PCT Int. Appl. (2018), WO 2018160889 Al 20180907; incorporated by reference], HATU (80 mg,

0.211 mmol), DIPEA (0.122 mL, 0.702 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0-100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(615 bromo-3-methylpyridin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (10b) (49 mg, 67% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î7₆) δ 10.29 (s, 1H, D2O exchangeable), 8.72 - 8.51 (m, 3H, 2H D2O exchangeable), 8.37 (s, 1H), 7.71 - 7.53 (m, 2H), 7.45 (d, J= 7.9 Hz, 1H), 7.38 (d, J= 8.5 Hz, 1H), 5.67 (d, J= 17.4 Hz, 1H), 5.38 (d, J = 17.3 Hz, 1H), 4.36 (dd, J= 9.2, 5.2 Hz, 1H), 3.66 (dd, J= 5.6, 2.4 Hz, 1H), 2.61 - 2.52 (m,

4H), 2.05 (d, J= 5.4 Hz, 1H), 2.01 (s, 3H), 1.33 (s, 3H), 1.14 - 0.91 (m, 2H); MS (ES+):

548.0 (M+l), 569.9 (M+Na); (ES-): 546.0 (M-l); Analysis calculated for C₂₆H26BrN7O2.3H2O.1.15HCl: C, 48.46; H, 5.19; Cl, 6.33; N, 15.21; Found: C, 48.64; H, 5.03; Cl, 6.07; N, 15.02.

Scheme 11

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (llf)

Step-1: Préparation of 2-amino-5-(trifluoromethyl)-lH-indole-3-carbonitrile (11b)

A mixture of 2,2,2-trifluoro-N-(2-iodo-4-(trifluoromethyl)phenyI)acetamide (lia) (4.00 g, 10.44 mmol), malononitrile (0.828 g, 12.53 mmol), L-proline (0.240 g, 2.089 mmol), Cul (0.199 g, 1.044 mmol), and K2CO3 (2.89 g, 20.89 mmol) were suspended in a 1:1 mixture of DMSO (20 mL) and H2O (20 mL) and stirred at 60 °C for 16 h under an argon atmosphère.

The reaction mixture was diluted with saturated NH4CI (30 mL) and extracted with EtOAc (30 mL x 3). The combined organics were washed with H2O (30 mL x 4), brine (30 mL), dried, filtered and concentrated in vacuum. The residue obtained was purified by flash column chromatography [S1O2 gel (40 g), eluting with EtOAc in hexane from 0-50%] to provide 2-amino-5-(trifluoromethyl)-lH-indole-3-carbonitrile (11b) (1.24 g, 53 % yield) as an orange solid; Ή NMR (300 MHz, DMSO-t/e) δ 11.14 (s, 1H), 7.37 (d, J = 1.7 Hz, 1H), 7.30 (d, J = 8.3 Hz, 1H), 7.22 (dd, J = 8.4, 1.8 Hz, 1H), 7.12 (s, 2H). ^,9F NMR (282 MHz, DMSO-J₆) δ -58.93; MS (ES+): 226 (M+l); (ES-): 224 (M-l).

Step-2: Préparation of 6-(trifluoromethyl)-9H-pyrimido[4,5-b]indoI-4-amine (11c)

Compound 11c was prepared according to the procedure reported in step-1 of scheme-6, from

2-amino-5-(trifhioromethyl)-lH-indole-3-carbonitrile (11b) (1.24 g, 5.51 mmol) using trimethyl orthoformate (18.12 mL, 165 mmol), AcOH (1.575 mL, 27.5 mmol) andNHiOAc (2.122 g, 27.5 mmol). This gave after workup and purification [S1O2 gel (24 g), eluting with

MeOH in DCM from 0-5%] 6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-4-amine (11c) (1.13 g, 4.48 mmol) as a pale yellow solid; Ή NMR (300 MHz, DMSO-^) δ 12.25 (s, 1H), 8.78 (s, 1H), 8.31 (s, 1H), 7.66 (dd, J = 8.4, 1.7 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.46 (s, 2H). ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -58.13; MS (ES+): 253 (M+l); (ES-): 251 (M-l).

Step-3 : Préparation of ZerZ-butyl 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido [4,5 -b] indol-9yl)acetate (lld)

Compound lld was prepared according to the procedure reported in step-1 of scheme-1, from 6-(trifluoromethyI)-9H-pyrimido[4,5-b]indol-4-amine (11c) (0.75 g, 2.97 mmol) in DMF (20 mL) using ZerZ-butyl 2-bromoacetate (0.580 g, 2.97 mmol), CS2CO3 (1.163 g, 3.57 10 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] ZerZ-butyl 2-(4amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (lld) (0.89 g, 82% yield) as a pale orange solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 8.83 (s, 1H), 8.36 (s, 1H), 7.84 - 7.70 (m, 2H), 7.60 (s, 2H), 5.21 (s, 2H), 1.41 (s, 9H).

Step-4: Préparation of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie)

Compound lie was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (lld) (0.89 g, 2.429 mmol) using TFA (4.43 g, 38.9 mmol) in DCM (7 mL) and stirring at RT for 16 h.

This gave after workup 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (1.26 g) TFA sait as an orange solid; ‘H NMR (300 MHz, DMSO-î/<î) δ 8.96 (s, 1H), 8.57 (s, 1H), 8.43 (s, 2H), 7.96 (d, J = 8.6 Hz, 1H), 7.84 (dd, J = 8.6, 1.7 Hz, 1H), 5.31 (s, 2H). ¹⁹F NMR (282 MHz, DMSO-^) δ -58.51, -74.65; MS (ES+): 311 (M+l); (ES-): 309 (M-l). '

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane3-carboxamide (llf)

Compound llf was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (50 mg, 0.118 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6-bromo-3methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (10a) (40.9 mg, 0.118 mmol), HATU (67.2 mg, 0.177 mmol), DIPEA (76 mg, 0.589 mmol) and stirring at RT

for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(trifluorômethyl)-9H-pyrimido[4,55 b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane3-carboxamide (1 If) (55 mg, 77% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-îZô) δ 10.29 (s, 1H, D₂O exchangeable), 8.99 (s, 1H), 8.59 (m, 3H, 2H D₂O exchangeable), 7.84 (d, J= 2.1 Hz, 2H), 7.62 (d, J= 8.0 Hz, 1H), 7.45 (d, J= 7.9 Hz, 1H), 5.73 (d, J= 17.4 Hz, 1H), 5.45 (d, J= 17.3 Hz, 1H), 4.36 (dd, J= 9.2, 5.2 Hz, 1H), 3.67 (dd,

J= 5.4, 2.5 Hz, 1H), 2.60-2.54 (m, 1H), 2.09-2.02 (m, 1H), 2.00 (s, 3H), 1.34 (s, 3H), 1.06 (t, J= 5.3 Hz, 1H), 0.99 (dd, J= 5.4, 2.4 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ 58.51; MS (ES+): 602.0 (M+l), 623.9 (M+Na); (ES-): 600.0 (M-l); Analysis calculated for C₂₆H₂₃BrF₃N₇O₂.2.5H₂O.lHCl: C, 45.66; H, 4.27; Cl, 5.18; N, 14.34; Found: C, 45.66; H, . 4.02; Cl, 5.29; N, 14.26. .

Scheme 12

nh₂

Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (12a)

Compound 12a was prepared according to the procedure reported in step-3 of scheme-1, from 20 TFA sait of 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (le) (50 mg, 0.14 mmol) in

DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (10a) (48.7 mg, 0.14 mmol), HATU (80 mg, 0.211 mmol), DIPEA (0.122 mL, 0.702 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in

DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%]

(lR,3S,5R)-2-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-3methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (12a) (47 mg, 63 % yield) as a white solid; ’H NMR (300 MHz, DMSO-tZ₆) δ 10.31 (s, 1H, D₂O exchangeable), 8.77 (s, 2H, D₂O exchangeable), 8.66 (s, 1H), 8.55 (d, J= 7.9 Hz, 1H), 7.71 (d, J= 8.2 Hz,

1H), 7.62 (d, 7= 8.0 Hz, 1H), 7.56 (t, J= 7.7 Hz, 1H), 7.45 (dd, J= 7.9, 6.0 Hz, 2H), 5.72 (d,

J= 17.3 Hz, 1H), 5.42 (d, J= 17.3 Hz, 1H), 4.37 (dd, J= 9.2, 5.3 Hz, 1H), 3.68 (dd, J= 5.6, 2.4 Hz, 1H), 2.61-2.52 (m, 1H), 2.09-2.01 (m, 1H), 2.00 (s, 3H), 1.34 (s, 3H), 1.06 (t, 7= 5.4 Hz, 1H), 0.99 (dd, J= 5.3, 2.4 Hz, 1H). MS (ES+): 534.0 (M+l), 556.0 (M+Na); (ES-): 532.0 (M-l); Analysis calculated for C₂₅H₂₄BrN7O₂.2.5H₂O.l.lHCl: C, 48.47; H, 4.90; Cl,

6.29; N, 15.83; Found: C, 48.70; H, 4.84; Cl, 6.26; N, 15.63.

Scheme 13

Préparation of (S)-l-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin2-yl)pyrrolidine-2-carboxamide (13b)

Compound 13b was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (le) (50 mg, 0.14 mmol) in DMF (2 mL) using TFA sait of (S)-N-(6-bromopyridin-2-yl)pyrrolidine-2carboxamide (13a) [(53.9 mg, 0.140 mmol); Wiles, Jason A. et al., PCT Int. Appl. (2017), WO 2017035353 Al 20170302], HATU (80 mg, 0.211 mmol), DIPEA (0.122 mL, 0.702 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (S)-l-(2-(4-amino-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (13b) (42 mg, 61% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-îZ₆) δ 11.23 and 10.80 (2s, 1H D₂O exchangeable), 8.63 - 8.50 (m, 3H, 2H D₂O exchangeable), 8.45 (d, J= 7.9 Hz, 1H), 8.11 and 7.93 (2d, 7=8.1 Hz, 1H), 7.64 (q, J= 8.2 Hz, 2H), 7.55-7.44 (m, 1H), 7.40-7.31

(m, 1H), 7.24 (d, J= 7.7 Hz, 1H), 5.45 - 5.28 (m, 2H), 4.46 (dd, J= 8.3, 4.3 Hz, 1H), 3.80 (tt, J= 9.7, 5.3 Hz, 2H), 2.23 - 1.76 (m, 4H); MS (ES+): 494.0 (M+l); (ES-): 492.0 (M-l);

Analysis calculated for C₂₂H₂oBrN₇02.2.75H₂0.1HCl: C, 45.53; H, 4.60; Cl, 6.11; N, 16.89;

Found: C, 45.78; H, 4.22; Cl, 6.37; N, 16.65.

Scheme 14

cf₃

Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (14a)

Compound 14a was prepared according to the procedure reported in step-3 of scheme-1, from 10 TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (50 mg, 0.118 mmol) in DMF (2 mL) using HCl sait of (2S,4R)-N-(6-bromo-3methylpyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (3a) (39.9 mg, 0.118 mmol), HATU (67.2 mg, 0.177 mmol), DIPEA (0.103 mL, 0.589 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting 75 with DMA-80 in DCM from 0 -100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yI)acetyl)-N(6-bromo-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (14a) (40mg, 57% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-ùfe) δ (a mixture of two rotamers) 10.89 and 10.46 (2s, 1H, D₂O exchangeable), 8.96 (s, 1H), 8.55 (s, 1H), 8.44 (s, 2H, D₂O exchangeable), 7.90 - 7.69 (m, 2H), 7.59 (d, J= 8.0 Hz, 1H), 7.42 (d, 8.0 Hz, 1H), 5.76 -

5.33 (m, 3H), 4.57 (t, J= 8.6 Hz, 1H), 4.33 (dd, J= 21.8, 12.7 Hz, 1H), 4.18 - 3.95 (m, 1H), 2.73 (s, 1H), 2.30 - 2.11 (m, 1H), 1.93 (s, 3H); ^,9F NMR (282 MHz, DMSO-î7₆) δ -58.47, 58.75, -176.20; MS (ES+): 594.0 (M+l); (ES-): 592.0 (M-l); Analysis calculated for

C₂4H₂oBrF₄N₇0₂.1.75H₂0.1HCl: C, 43.52; H, 3.73; Cl, 5.35; N, 14.80; Found: C, 43.54; H,

3.24; Cl, 4.96; N, 14.54.

Scheme 15

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide 5 (15a)

Compound 15a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (lie) (50 mg, 0.118 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6-bromo-3methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (Id) (39.2 mg, 0.118 mmol),

HATU (67.2 mg, 0.177 mmol), DIPEA (0.103 mL, 0.589 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol15 9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (15a) (47 mg, 68% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 10.27 (s, 1H, D.O exchangeable), 9.00 (s, 1H), 8.70 - 8.54 (m, 3H, 2H D.O exchangeable), 7.93 7.80 (m, 2H), 7.62 (d, J= 8.0 Hz, 1H), 7.45 (d, J= 7.9 Hz, 1H), 5.78 (d, J= 17.4 Hz, 1H), 5.51 (d, J= 17.3 Hz, 1H), 4.38 (dd, J= 9.2, 5.1 Hz, 1H), 3.90 (td, J= 6.3, 5.4, 2.3 Hz, 1H),

2.46 - 2.18 (m, 2H), 2.03 - 1.86 (m, 4H), 1.11 (dt, J = 9.4, 5.3 Hz, 1H), 0.83 (dt, J= 7.1, 3.4

Hz, 1H); ^I9F NMR (282 MHz, DMSO-dfe) δ -58.52; MS (ES+): 587.9 (M+l); (ES-): 586.0 (M-l); Analysis calculated for C2₅H2iBrF₃N₇O2.HC1.1.75H2O: C, 45.75; H, 3.92; Cl, 5.40; N, 14.94; Found: C, 45.96; H, 3.65; Cl, 4.95; N, 14.71.

Scheme 16

I

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (16e)

Step-1: Préparation of 7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-4-amine (16b)

Compound 16b was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-6-(trifluoromethyl)-lH-indole-3-carbonitrile (16a) (1.95 g, 8.66 mmol; CAS # 1242140-69-7) using trimethyl orthoformate (18.95 mL, 173 mmol), AcOH (2.476 mL, 43.3 mmol) and NFLOAc (3.34 g, 43.3 mmol). This gave after workup 7-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-4-amine (16b) (2.08 g, 77% yield) acetic acid sait as a pale yellow solid; *H NMR (300 MHz, DMSO-î/ô) δ 12.20 (s, 1H), 11.98 (s, 1H), 8.52 (d, J= 8.3 Hz, 1H), 8.31 (s, 1H), 7.76 - 7.65 (m, 1H), 7.58 - 7.47 (m, 1H), 7.43 (s, 2H), 1.91 (s, 3H); MS (ES+): 253.10 (M+l).

Step-2: Préparation of tert-butyl 2-(4-amino-7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (16c)

Compound 16c was prepared according to the procedure reported in step-1 of scheme-1, from 7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-4-amine (16b) (2 g, 7.93 mmol) in DMF (50 mL) using CS2CO3 (3.10 g, 9.52 mmol). This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with EtOAc in Hexane from 0-100%] tertbutyl 2-(4-amino-7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (16c) (0.93 g,

32% yield) as a pale yellow solid; 'H NMR (300 MHz, DMSO-ri₆) δ 8.57 (d, J= 8.2 Hz, 1H),

8.36 (s, 1H), 8.06 (d, J= 1.5 Hz, 1H), 7.65 - 7.51 (m, 3H), 5.25 (s, 2H), 1.40 (s, 9H); MS (ES+): 367.10 (M+l).

Step-3 : Préparation of 2-(4-amino-7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (16d)

Compound 16d was prepared according to the procedure reported in step-2 of scheme-1, from terZ-butyl 2-(4-amino-7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (16c) (0.92 g, 2.51 mmol) using TFA (14.41 mL, 37.7 mmol; 20% TFA in DCM) and stirring at RT for 16 h. This gave after workup 2-(4-ammo-7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (16d) (0.9 g, 84% yield) TFA sait as a white solid; ’H NMR (300 MHz,

DMSO-dô) δ 8.64 (d, J= 8.3 Hz, 1H), 8.47 (s, 1H), 8.21 (s, 1H), 8.02 (s, 2H), 7.67 (d, J= 8.3 Hz, 1H), 5.31 (s, 2H); MS (ES+): 311.00 (M+l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-7-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (16e)

Compound 16e was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (16d) (75 mg, 0.177 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (70.0 mg, 0.177 mmol), HATU (101 mg, 0.265 mmol), DIPEA (0.154 mL, 0.884 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (16e) (62 mg,

61% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î7₆) δ 10.76 (s, 1H, D₂O exchangeable), 8.73 (d, J= 8.1 Hz, 3H, 2H D₂O exchangeable), 8.65 (s, 1H), 8.17 (s, 1H), 8.00 (d, J= 8.1 Hz, 1H), 7.77 - 7.61 (m, 2H), 7.30 (d, J= 7.7 Hz, 1H), 5.87 (d, J= 17.4 Hz, 1H), 5.51 (d, J= 17.3 Hz, 1H), 4.42 (dd, J= 9.0, 5.5 Hz, 1H), 3.91 (ddd, J= 7.4, 5.4, 2.3 Hz, 1H), 2.41 -2.13 (m, 2H), 1.92 (t, J= 6.4 Hz, 1H), 1.09 (dt, J= 8.7, 5.5 Hz, 1H), 0.77 (td, J=

5.2, 2.3 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-dfe) δ -59.33; MS (ES+): 574.0 (M+l); (ES-):

572.0 (M-l); Analysis calculated for C₂4Hi9BrF₃N₇O₂.1.5H₂O.lHCl: C, 45.19; H, 3.63; Cl, 5.56; N, 15.37; Found: C, 45.06; H, 3.61; Cl, 5.38; N, 15.36.

Préparation of ieri-butyl (4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-7-yl)carbamate (17f)

Step-1 : Préparation of 7-bromo-9H-pyrimido[4,5-b]indol-4-amine (17b)

Compound 17b was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-6-bromo-lH-indole-3-carbonitrile (17a) (2 g, 8.47 mmol; CAS # 1427028-361) using trimethyl orthoformate (18.54 mL, 169 mmol), AcOH (2.423 mL, 42.4 mmol) and NH4OAC (3.27 g, 42.4 mmol). This gave after workup 7-bromo-9H-pyrimido[4,5-b]indol-410 amine (17b) (2.29 g, 84% yield) as a pale yellow solid; *H NMR (300 MHz, DMSO-ùfe) δ

11.97 (s, 2H), 8.27 (t, J= 4.2 Hz, 2H), 7.58 (d, J= 1.8 Hz, 1H), 7.36 (dd, J= 8.4, 1.8 Hz, 1H), 7.26 (s, 2H), 1.92 (s, 3H).

Step-2: Préparation of Zer/-butyl 2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (17c)

Compound 17c was prepared according to the procedure reported in step-2 of scheme-16, from 7-bromo-9H-pyrimido[4,5-b]indol-4-amine (17b) (2.2 g, 6.81 mmol) in DMF (75 mL) using ter/-butyl 2-bromoacetate (1.056 mL, 7.15 mmol), CS2CO3 (4.88 g, 14.98 mmol) and stirring at RT for 15 h. Additional /ert-butyl 2-bromoacetate (1.006 mL, 6.81 mmol) and K2CO3 (0.941 g, 6.81 mmol) were needed for completion of reaction. This gave after workup and purification by flash column chromatographÿ [silica gel (40 g), eluting with EtOAc in hexane from 0-100%] fôrZ-butyl 2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (17c) (1.25 g, 49% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-ri₆) δ 8.31 (t, J = 4.2 Hz, 2H), 7.91 (d, J= 1.7 Hz, 1H), 7.46 - 7.36 (m, 3H), 5.14 (s, 2H), 1.41 (s, 9H).

Step-3: Préparation of ZerZ-butyl 2-(4-amino-7-((tert-butoxycarbonyl)amino)-9Hpyrimido[4,5-b]indol-9-yl)acetate (17d)

To a degassed solution of ZerZ-butyl 2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9yl)acetate (17c) (200 mg, 0.530 mmol) in toluene (10 mL) was added XPhos (50.5 mg, 0.106 mmol), t-butyl carbamate (93 mg, 0.795 mmol), Pdz(dba)3 (48.5 mg, 0.053 mmol) and césium carbonate (173 mg, 0.530 mmol), filled with nitrogen and heated at 90 °C for 16 h. The reaction mixture was diluted with ethyl acetate (100 mL), washed with water (50 mL), brine (50 mL), dried and concentrated in vacuum. The obtained residue was purified by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0-50%] to give ZerZ-butyl 2-(4-amino-7-(tert-butoxycarbonylamino)-9H-pyrimido[4,5-b]indol-9yl)acetate (17d) (92mg, 42% yield) as a yellow solid; MS (ES+): 414.20 (M+l).

Step-4: Préparation of 2-(4-amino-7-((ZerZ-butoxycarbonyl)amino)-9H-pyrimido[4,5-b]indol15 9-yl)acetic acid (17e)

To a stirred solution of ZerZ-butyl 2-(4-amino-7-(ZerZ-butoxycarbonylamino)-9Hpyrimido[4,5-b]indol-9-yl)acetate (17d) (90 mg, 0.218 mmol) in THF/MeOH (4 mL; ratio 1:1) was added lithium hydroxide hydrate (1.088 mL, 1.088 mmol) and stirred atRT for 15 h. The reaction mixture was concentrated in vacuo, diluted with water (1 mL), acidified to pH

5-7 using IM HCl and concentrated to dryness to afford 2-(4-amino-7-((ZerZbutoxycarbonyl)amino)-9H-pyrimido[4,5-b]indol-9-yI)acetic acid (17e) (67 mg, 86 % yield); 'H NMR (300 MHz, DMSO-J₆) δ 9.71 (s, 1H), 8.63 - 8.45 (m, 4H), 8.38 (d, J= 8.6 Hz, 1H), 7.95 (s, 1H), 7.36 (dd, J= 8.7, 1.8 Hz, 1H), 5.19 (s, 2H), 1.51 (s, 9H); MS (ES+): 358.10 (M+l); (ES-): 356.10 (M-l).

Step-5: Préparation of ZerZ-butyl (4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-7yl)carbamate (17f)

Compound 17f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-((ZerZ-butoxycarbonyI)amino)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (17e) (60mg, 0.168 mmol) in DMF (1 mL) using TFA sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (66.5 mg, 0.168 mmol),

HATU (96 mg, 0.252 mmol) DIPEA (0.146 mL, 0.839 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] tert-butyl 4-amino-9-(2-((lR,3S,5R)-3-(6bromopyridin-2-ylcarbamoyl)-2-azabicyclo[3.1.0]hexaii-2-yl)-2-oxoethyl)-9H-pyrimido[4,5b]indol-7-ylcarbamate (17f) (48mg, 46% yield). 10 mg of this compound was purified by reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] to yield teri-butyl 4-amino-9-(2-((lR,3S,5R)-3-(6bromopyridin-2-ylcarbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5b]indoI-7-ylcarbamate (17f) (6 mg) HCl sait as a white solid; Ή NMR (300 MHz, MeOD-i/4) δ 8.46 (s, 1H), 8.19 (d, J= 8.7 Hz, 1H), 8.05 (d, J= 8.2 Hz, 1H), 8.00 (d, J= 1.8 Hz, 1H), 7.60 (t, J= 8.0 Hz, 1H), 7.35 (dd, J= 8.7, 1.9 Hz, 1H), 7.24 (dd, J= 7.8, 0.7 Hz, 1H), 5.74 (d, J= 17.2 Hz, 1H), 5.53 (d, J= 17.1 Hz, 1H), 4.56 (t, J= 7.1 Hz, 1H), 3.89 (ddd, J= ΊΑ, 5.4, 2.4 Hz, 1H), 2.46 (dd, J= 7.9, 3.6 Hz, 2H), 2.04 (d, J= 6.4 Hz, 1H), 1.56 (s, 9H), 1.24 (dt, J = 8.6, 5.5 Hz, 1H), 1.00 (dt, J= 7.6, 3.7 Hz, 1H). MS (ES+): 621.2 (M+l); (ES-): 619.1 (M1).

Scheme 18

Préparation of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-(2-(4,7-diamino-9H-pyrimido[4,5b]indol-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (18a)

Compound 18a was prepared according to the procedure reported in step-3 of scheme-1, from terLbutyl (4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-7-yl)carbamate (17f) (35mg, 0.056 mmol) in DCM (1 mL) using 2,2,2-trifluoroacetic acid (0.087 mL, 1.126 mmol) and stirring at RT for 16 h. This gave after workup and purification twice by reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1%

100

HCl) from 0-100%] (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-(2-(4,7-diamino-9Hpynmido[4,5-b]indol-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (17 mg, 58% yield) (18a) HCl sait as a white solid. Ή NMR (300 MHz, DMSO-î/₆) δ 10.74 (s, 1H, D₂O exchangeable), 8.45 (s, 1H), 8.29 (s, 2H, D₂O exchangeable), 8.18 (d, J= 8.6 Hz, 1H), 7.95 (d, J= 8.2 Hz, 1H), 7.64 (t, J= 7.9 Hz, 1H), 7.25 (d, J= 7.7 Hz, 1H), 6.81 (d, J= 7.0 Hz, 2H), 5.58 (d, J= 17.4 Hz, 1H), 5.22 (d, J= 17.4 Hz, 1H), 4.36 (dd, J= 9.0, 5.4 Hz, 1H), 3.89 -3.76 (m, 1H), 2.34-2.06 (m, 2H), 1.85 (d, J = 7.7 Hz, 1H), 1.10-0.92 (m, 1H), 0.68(d,J = 5.9 Hz, 1H). MS (ES+): 521.1 (M+l); (ES-): 519.1 (M-l).

Préparation of (2S,4R)-l-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (19a)

Compound 19a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yI)acetic acid (le) (150 mg, 0.421 mmol) in DMF (5 mL) using TFA sait of (2S,4R)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2carboxamide (5d) (169 mg, 0.421 mmol), HATU (192 mg, 0.505 mmol), DIPEA (272 mg, 2.105 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2carboxamide (19a) (62 mg, 61% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-îZô) δ (a mixture of two rotamers) 11.37 (s) and 10.98 (s) (2s, 1H, D2O exchangeable),, 8.80 - 8.56 (m, 3H, 2H D2O exchangeable), 8.51 (d, J= 7.9 Hz, 1H), 7.98 (d, J= 8.1 Hz, 1H), 7.75 - 7.63 (m, 2H), 7.55 (t, J= 7.7 Hz, 1H), 7.50 - 7.37 (m, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.64 (d, 3.5 Hz, 1H), 5.60 - 5.45 (m, 1H), 5.40 (d, J= 17.3 Hz, 1H),

101

4.71-4.57 (m, 1H), 4.43-4.21 (m, 1H), 4.18-3.96 (m, 1H), 2.74-2.50 (m, 1H), 2.292.02 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-J₆) δ -175.73, -176.14; MS (ES+): 512.0 (M+l), (ES-): 510.0 (M-l); Analysis calculated for C₂₂Hi9BrFN₇O₂.HC1.2.5H₂O: C, 44.50; H, 4.24; Cl, 5.97; N, 16.51; Found: C, 44.36; H, 4.10; Cl, 5.76; N, 16.25.

Scheme 20

Préparation of (2S,4R)-l-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoropynOlidine-2-carboxamide (20a)

Compound 20a was prepared according to the procedure reported in step-3 of scheme-1, from 10 TFA sait of 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (le) (200 mg, 0.561 mmol) in DMF (10 mL) using TFA sait of (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine2-carboxamide (7a) (218 mg, 0.561 mmol), HATU (256 mg, 0.674 mmol), DIPEA (0.490 mL, 2.81 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse-phase column chromatography [C-l 8 column, 100 g, eluting with 0.1% aqueous HCl in H₂O and MeCN from 0-100%] (2S,4R)-l-(2-(4-amino-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2carboxamide (20a) (121 mg, 43% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-dô) (a mixture of two rotamers) δ 9.15 (t, J= 5.8 Hz) and 8.79 - 8.64 (m, 3H, D₂O exchangeable), 8.61 (d, J= 3.0 Hz, 1H), 8.59 - 8.50 (m, 1H), 7.74 (d, J= 8.1 Hz, 1H), 7.58 7.34 (m, 4H), 7.21 - 7.11 (m, 1H), 6.92 - 6.79 (m, 1H), 5.72 - 5.53 (m, 1H), 5.54 - 4.76 (m, 2H), 4.56-4.14 (m, 4H), 4.14-3.92 (m, 1H), 2.62-2.40 (m, 1H), 2.22-1.95 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-cfc) δ -121.24, -121.76, -176.25, -176.47; MS (ES+): 499/501 (M+l).

Scheme 21

Préparation of (2S,4R)-l-(2-(4-amino-6-methyI-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (21a)

Compound 21a was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (6d) (50 mg,

0.135 mmol) in DMF (5 mL) using TFA sait of (2S,4R)-N-(6-bromopyridin-2-yl)-4fluoropyrrolidine-2-carboxamide (5d) (54.3 mg, 0.135 mmol), HATU (61.6 mg, 0.162 mmol), DIPEA (87 mg, 0.675 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0-5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-1(2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4fluoropyrrolidine-2-carboxamide (21a) (35 mg, 49% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-cfc) δ (a mixture of rotamers) 11.35 (s) and 10.97 (s, 1H, D2O exchangeable), 8.76 - 8.52 (m, 3H, 2H D2O exchangeable), 8.35 (s, 1H), 7.98 (d, J= 8.2 Hz, 1H), 7.69 (t, J= 7.9 Hz, 1H), 7.60 (d, J= 8.4 Hz, 1H), 7.38 (d, J= 8.5 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.71 - 5.25 (m, 3H), 4.62 (t, J= 8.5 Hz, 1H), 4.37 - 4.23 (m, 1H), 4.16 - 3.96 (m, 1H), 2.66 - 2.43 (m, 4H), 2.30 - 1.95 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-Je) δ 175.73, -176.15; MS (ES+): 526.0 (M+l), (ES-): 524.0 (M-l); Analysis calculated for

C23H₂iBrFN₇O2.LlHC1.2.25H2O: C, 45.51; H, 4.42; Cl, 6.42; N, 16.15; Found: C, 45.21; H, 4.47; Cl, 6.40; N, 15.88.

Scheme 22

103

6b ^22a 22b

HATU, D1PEA

22c (-)-isomer

22d (+)-isomer

Préparation of (lR,3S,5R)-2-((-)-2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9yl)propanoyl)-N-(6-bromopyridin-2-yl)-2-azabicycIo[3.1.0]hexane-3-carboxamide (22c) and (lR,3S,5R)-2-((+)-2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)propanoyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (22d)

Step-1: Préparation of tert-butyl 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9yl)propanoate (22a)

Compound 22a was prepared according to the procedure reported in step-1 of scheme-1, from 10 6-methyl-9H-pyrimido[4,5-b]indol-4-amine (6b) (200 mg, 1.009 mmol) in DMF (10 mL) using (R)-tert-butyl 2-bromopropanoate (253 mg, 1.211 mmol; CAS # 54631-38-8), CS2CO3 (657 mg, 2.018 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0-3%] a mixture of two enantiomers of tert-butyl 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-915 yl)propanoate (22a) (250 mg, 76% yield) as a pale yellow solid; *H NMR (300 MHz, DMSOd₆) δ 8.26 (d, J = 6.0 Hz, 1H), 8.23 - 8.15 (m, 1H), 7.40 (dd, J = 7.4, 3.1 Hz, 1H), 7.30 - 7.07 (m, 3H), 5.81 - 5.63 (m, 1H), 2.46 (s, 3H), 1.66 (d, J = 7.1 Hz, 3H), 1.30 (d, J = 5.7 Hz, 9H); MS (ES+): 327 (M+l).

104

Step-2: Préparation of 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)propanoic acid (22b)

Compound 22b was prepared according to the procedure reported in step-2 of scheme-1, from Zeri-butyl 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)propanoate (22a) (250 mg, 0.766 mmol) in DCM (20 mL) using TFA (0.879 mL, 11.49 mmol) and stirring at RT for 16 h. This gave after workup a mixture of two enantiomers of 2-(4-amino-6-methyl-9Hpyrimido[4,5-b]indol-9-yl)propanoic acid (22b) (304 mg) TFA sait as a white solid; *HNMR (300 MHz, DMSO-î/₆) δ 8.55 (s, 1H), 8.49 - 8.36 (m, 2H), 8.36 (s, 1H), 7.62 (d, J= 8.5 Hz, 1H), 7.37 (dd, J= 8.6, 1.6 Hz, 1H), 5.86 (q, J=7.1 Hz, 1H), 1.75 (d, 7 = 7.2 Hz, 3H); ¹⁹F

NMR (282 MHz, DMSOW) δ -74.15; MS (ES+): 271 (M+l), (ES-): 269 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-((-)-2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9yl)propanoyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1,0]hexane-3-carboxamide (22c) and (lR,3S,5R)-2-((+)-2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)propanoyl)-N-(6bromopyridin-2-yl)-2-azabicyclo [3.1.0]hexane-3-carboxamide (22d)

Compounds 22c and 22d were prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)propanoic acid (22b) (50 mg, 0.130 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (41.5 mg, 0.130 mmol), HATU (59.4 mg, 0.156 mmol), DIPEA (84 mg, 0.651 mmol) and stirring at RT for 16 h. This 20 gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%].

1. (lR,3S,5R)-2-((-)-2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)propanoyl)-N25 (6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (22c) (17 mgs,

25%) HCI sait as a white solid; Ή NMR (300 MHz, DMSOW) δ 10.80 (s, 1H, D₂O exchangeable), 8.64 (s, 1H), 8.62 - 8.50 (m, 2H, D₂O exchangeable), 8.40 (s, 1H), 8.08 (d, J = 8.1 Hz, 1H), 7.76 (t, J = 7.9 Hz, 1H), 7.56 (d, J = 8.5 Hz, 1H), 7.37 (dd, J = 7.9, 2.6 Hz, 2H), 6.28 (q, J = 6.8 Hz, 1H), 4.27 (t, J = 7.2 Hz, 1H), 3.27 - 3.24 (m,

1H), 2.04 - 1.95 (m, 2H), 1.61 (m, 3H), 1.57 - 1.47 (m, 1H), -0.36 (m, 1H), -1.03 (m,

1H); MS (ES+): 534/536 (M+l), (ES-): 532/534 (M-l); Chiral HPLC: AD-H column 80/20 [(0.1% DEA in n-Hexane in 0.1% DEA in éthanol)] 1.0 mL/min UV détection

105

245 nm, 30 mins run time (Temp 40 °C). Rt= 10.54 (peak-1 (22c), 98.0742 %); Rt = 19.473 (peak-2; (22d) 1.388%) 97.4084% ee; Optical rotation [œ]d = -224 (c = 0.1,

MeOH)

2. (lR,3S,5R)-2-((+)-2-(4-amino-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)propanoyl)-N5 (6-bromopyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (22d) (22 mgs,

32%) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.64 (s, 1H, D₂O exchangeable), 8.62 (s, 1H), 8.59 - 8.46 (m, 2H, D₂O exchangeable), 8.36 (s, 1H), 7.92 (d, J = 8.2 Hz, 1H), 7.72 (d, J = 7.9 Hz, 1H), 7.66 (d, J = 8.3 Hz, 1H), 7.34 (d, J = 7.7 Hz, 1H), 7.29 (d, J = 8.6 Hz, 1H), 6.22 (q, J = 7.0 Hz, 1H), 4.53 (dd, J = 9.1, 5.6

Hz, 1H), 3.06 - 2.96 (m, 1H), 2.30 - 2.17 (m, 1H), 2.07 (m, 1H), 1.68 (m, 4H), 0.88 (m, 1H), 0.77 (m, 1H); MS (ES+): 534/536 (M+l), (ES-): 532/534 (M-l); Chiral HPLC: AD-H column 80/20 [(0.1% DEA in n-Hexane in 0.1% DEA in éthanol)] 1.0 mL/min UV détection 245 nm, 30 mins run time (Temp 40 °C); Rt= 10.54 (peak-1 (22c) 0 %); R_t = 19.427 (peak-2; (22d) 100%) >99.99% ee; Optical rotation [œ]d = +92.632 (c = 0.095, MeOH).

Scheme 23

106

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (23 f)

Step-1: Préparation of 2-amino-6-methyl-lH-indole-3-carbonitriIe (23b)

Compound 23b was prepared according to the procedure reported in step-1 of scheme-11, 5 from 2,2,2-trifluoro-N-(2-iodo-5-methyIphenyl)acetamide (23a) (7.03 g, 21.36 mmol) in

DMSO (20 mL) using malononitrile (1.694 g, 26.6 mmol), L-proline (0.492 g, 4.27 mmol), Cul (0.407 g, 2.136 mmol), K2CO3 (5.91 g, 42.7 mmol) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [SiO₂ gel (40 g), eluting with EtOAc in hexane from 0-40%] 2-amino-6-methyl-lH-indole-3-carbonitrile (23b) (2.65 g, 10 73% yield) as an brown solid; *H NMR (300 MHz, DMSO-î/₆) δ 10.58 (s, 1H), 7.00 (d, J=

7.8 Hz, 1H), 6.93 (s, 1H), 6.78 (dd, J= 8.3, 1.5 Hz, 1H), 6.63 (s, 2H), 2.31 (s, 3H); MS (ES+): 172 (M+l); (ES-): 170 (M-l).

Step-2: Préparation of 7-methyl-9H-pyrimido[4,5-b]indol-4-amine (23c)

Compound 23c was prepared according to the procedure reported in step-1 of scheme-6, from 75 2-amino-6-methyl-lH-indole-3-carbonitrile (23b) (2.65 g, 15.48 mmol) using triethyl orthoformate (51.5 mL, 310 mmol), AcOH (4.43 mL, 77 mmol) andNHiOAc (5.97 g, 77 mmol). This gave after workup and purification [S1O2 gel (24 g), eluting with MeOH in DCM from 0-5%] 7-methyl-9H-pyrimido[4,5-b]indol-4-amine (23c) (1.45 g) as a brown yellow solid; 'H NMR (300 MHz, DMSO-ifc) δ 11.70 (s, 1H), 8.21 (s, 1H), 8.16 (d, J = 7.9

Hz, 1H), 7.23 (s, 1H), 7.15 - 6.96 (m, 3H), 2.45 (s, 3H); MS (ES+): 199 (M+l); (ES-): 197 (M-l).

Step-3: Préparation of fôri-butyl 2-(4-amino-7-methyl-9H-pyrimido[4,5-b]indol-9-yI)acetate (23d)

Compound 23d was prepared according to the procedure reported in step-1 of scheme-1, 25 from 7-methyl-9H-pyrimido[4,5-b]indol-4-amine (23c) (1.45 g, 7.31 mmol) in DMF (25 mL) using terributyl 2-bromoacetate (1.427 g, 7.31 mmol) CS2CO3 (2.86 g, 8.78 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] ter/-butyl 2-(4amino-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (23d) (0.84 g, 37% yield) as a pale yellow solid; ‘H NMR (300 MHz, DMSO-îZ₆) δ 8.26 (s, 1H), 8.22 (d, J= 8.0 Hz, 1H), 7.36 (s, 1H), 7.19 (s, 2H), 7.14 - 7.06 (m, 1H), 5.07 (s, 2H), 2.48 (s, 3H), 1.41 (s, 9H); MS (ES+): 313 (M+l).

107

Step-4: Préparation of 2-(4-amino-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (23e)

Compound 23e was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-7-methyl-9H-pyrimido[4,5-b]indol-9-yI)acetate (23d) (0.84 g, 2.69 mmol) using TFA (6.13 g, 53.8 mmol) in DCM (20 mL) and stirring at RT for 16 h. This gave after workup 2-(4-amino-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (23e) (1.01 g) TFA sait as a yellow solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.51 (s, 1H), 8.36 (d, J= 8.0 Hz, 1H), 8.24 (s, 2H), 7.59 (s, 1H), 7.26 (d, 1H), 5.21 (s, 2H), 2.49 (s, 3H); ^I9FNMR (282 MHz, DMSO-J₆) δ -74.09; MS (ES+): 257 (M+l), (ES-): 255 (M-l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3 -carboxamide (23f) Compound 23f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (23e) (50 mg, 0.135 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-215 azabicyclo[3.1.0]hexane-3-carboxamide (4a) (43.0 mg, 0.135 mmol), HATU (61.6 mg, 0.162 mmol), DIPEA (87 mg, 0.675 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (23f) (44 mg, 63% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ 10.76 (s, 1H, D2O exchangeable), 8.57 (s, 1H), 8.44 (s, 2H, D₂O exchangeable), 8.37 (d, J= 8.1 Hz, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.51 (s, 1H), 7.32 (d, J= 7.7 Hz, 1H), 7.29 - 7.23 (m, 1H), 5.71 (d, J = 17.3 Hz, 1H), 5.38 (d, J= 17.3 Hz, 1H), 4.42 (dd, J= 9.1, 5.6 Hz, 1H), 3.96 - 3.87 (m, 1H),

2.49 (s, 3H), 2.40 - 2.27 (m, 1H), 2.27-2.11 (m, 1H), 2.00-1.85 (m, 1H), 1.12-1.02 (m, 1H), 0.82 - 0.73 (m, 1H); MS (ES+): 520.0 (M+l), 518.0 (M-l); Analysis calculated for C24H22BrN7O2.L2HCL2.5H2O: C, 47.32; H, 4.67; Cl, 6.98; N, 16.10; Found: C, 47.10; H, 4.59; Cl, 6.75; N, 15.97.

Scheme 24

108

23e 24a

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (24a)

Compound 24a was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(4-amino-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (23e) (50 mg,

0.135 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicycIo[3.1.0]hexane-3-carboxamide (8a) (44.9 mg, 0.135 mmol), HATU (61.6 mg, 0.162 mmol), DIPEA (87 mg, 0.675 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with

MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (24a) (36 mg, 50% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-^) δ 10.77 (s, 1H, D₂O exchangeable), 8.61 (s, 3H, 2H D₂O exchangeable), 8.39 (d, J= 8.1 Hz, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.52 (s, 1H), 7.31 (d, J= 7.7 Hz, 1H), 7.26 (d, J= 8.9 Hz, 1H), 5.68 (d, J= 17.3 Hz, 1H), 5.34 (d, J= 17.3 Hz, 1H), 4.37 (dd, J= 9.1, 5.9 Hz, 1H), 3.69 (dd, J= 5.5, 2.4 Hz, 1H), 2.55-2.41 (m, 4H), 1.98 (dd, J= 13.2, 5.9 Hz, 1H), 1.31 (s, 3H), 1.08 - 0.97 (m, 1H), 0.97 - 0.89 (m, 1H); MS (ES+) 534.0 (M+l), 532.0 (M-l); Analysis calculated for C₂₅H₂₄BrN₇O₂.HC1.2.25H₂O: C, 49.11; H, 4.86; Cl, 5.80; N, 16.04; Found: C, 48.98; H, 4.79; Cl, 6.02; N, 15.90.

Scheme 25

109

11e 25a

Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(3-chlorô-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (25a)

Compound 25a was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (50 mg, 0.118 mmol) in DMF (10 mL) using TFA sait of (2S,4R)-N-(3-chloro-2fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (7a) (45.8 mg, 0.118 mmol), HATU (53.8 mg, 0.141 mmol), DIPEA (0.103 mL, 0.589 mmol) and stirring atRT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with 10 MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCI) from 0-100%] (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (25a) (28 mg, 42% yield) HCl sait as a white solid; *HNMR (300 MHz, DMSO-cfe) δ (a mixture of two 15 rotamers) 8.98 (d, J= 3.6 Hz, 1H), 8.71 - 8.52 (m, 4H, 2H D₂O exchangeable), 7.88 (d, J =

8.6 Hz, 1H), 7.75 (dd, J=8.7, 1.7 Hz, 1H), 7.45-7.31 (m, 1H), 7.17-7.08 (m, 1H), 6.80 (t, J= 7.9 Hz, 1H), 5.64 (d, J= 17.4 Hz, 1H), 5.43 (d, J= 4.0 Hz, 1H), 4.54 - 4.26 (m, 3H), 4.23 (d, J=5.8Hz, 1H), 4.19-4.05 (m, 1 H), 3.96 (dd, J= 12.3, 2.9 Hz, 1H), 2.62-2.41 (m, 1H), 2.23 - 1.91 (m, 1H); ¹⁹F NMR (282 MHz, DMSO) δ -58.53, -121.26, -121.72, -176.26, 20 176.42; MS (ES+): 567.0 (M+l), (ES-): 565.0 (M-l).

110

Scheme 26

Préparation of (2S,4R)-1 -(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (26a)

Compound 26a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (50 mg, 0.118 mmol) in DMF (5 mL) using TFA sait of (2S,4R)-N-(6-bromopyridin-2-yl)-4. fluoropyrrolidine-2-carboxamide (5d) (47.4 mg, 0.118 mmol), HATU (53.8 mg, 0.141 mmol), DIPEA (76 mg, 0.589 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-1(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2yl)-4-fluoropyrrolidine-2-carboxamide (26a) (33 mg, 48% yield) HCl sait as a white solid; *H

NMR (300 MHz, DMSOY) δ 10.97 (s, 1H, D₂O exchangeable), 8.95 (s, 1H), 8.64 - 8.38 (m, 3H, 2H D₂O exchangeable), 7.98 (d, J= 8.2 Hz, 1H), 7.84 (dd, J= 8.4, 5.0 Hz, 2H), 7.68 (t, J= 8.0 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.71 - 5.57 (m, 1H), 5.51 - 5.39 (m, 1H), 4.62 (dd, J= 9.7, 7.5 Hz, 1H), 4.32 (dd, J= 22.1, 12.6 Hz, 1H), 4.18 - 4.08 (m, 1H), 4.01 (dd, J= 12.8, 3.0 Hz, 1H), 2.68 - 2.33 (m, 1H), 2.30 - 1.97 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-îZô) δ -58.51, -175.70; MS (ES+): 580.0 (M+l), (ES-): 578.0 (M-l); Analysis calculated for C₂3Hi₈BrF₄N₇O₂.HC1.2.5H₂O: C, 41.74; H, 3.66; Cl, 5.36; N, 14.81; Found: C, 41.66; H, 3.49; Cl, 5.10; N, 14.74.

Scheme 27

111

11e 27a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyI)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (27a)

Compound 27a was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (50 mg, 0.118 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (37.5 mg, 0.118 mmol), HATU (53.8 mg, 0.141 mmol), DIPEA (76 mg, 0.589 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with

MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (27a) (42 mg, 62% yield) HCl sait as a white solid; ‘H NMR (300 MHz, DMSO-70 δ 10.75 (s, 1H, D₂O exchangeable), 8.96 (s, 1H), 8.59 (s, 1H), 8.52 (s, 2H, D₂O exchangeable), 8.00 (d, J= 8.2 Hz, 1H), 7.85 (s, 2H), 7.70 (t, J= 8.0 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.80 (d, J= 17.4 Hz, 1H), 5.46 (d, J= 17.3 Hz, 1H), 4.41 (dd, J= 9.1, 5.5 Hz, 1H), 3.95 - 3.88 (m, 1H), 2.40 2.28 (m, 1H), 2.28-2.13 (m, 1H), 1.99-1.82 (m, 1H), 1.14-1.00 (m, 1H), 0.81-0.74 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-d₆) δ -58.47; MS (ES+): 574.0 (M+l); (ES-): 572.0 (M-l);

Analysis calculated for C₂4Hi9BrF₃N7O₂.HC1.2H₂O: C, 44.56; H, 3.74; Br, 12.35; Cl, 5.48; N, 15.16; Found: C, 44.27; H, 3.69; Cl, 5.59; N, 14.75.

Scheme 28

112

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yI)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (28a)

Compound 28a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (50 mg, 0.118 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (39.2 mg, 0.118 mmol), HATU (53.8 mg, 0.141 mmol), DIPEA (76 mg, 0.589 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (28a) (50 mg, 72% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-7₆) δ 10.76 (s, 1H, D2O exchangeable), 8.96 (s, 1 H), 8.71 - 8.43 (m, 3H, 2H D₂O exchangeable), 8.01 (d, 7= 8.2 Hz, 1H), 7.91-7.80 (m, 2H), 7.69 (t, J= 8.0 Hz, 1H), 7.31 (d, 7= 7.7 Hz, 1 H), 5.76 (d, J= 17.4 Hz, 1H), 5.41 (d, J= 17.3 Hz, 1H), 4.36 (dd, J= 9.1, 6.0 Hz, 1H), 3.69 (dd, J= 5.5, 2.4 Hz, 1H), 2.52-2.39 (m, 1H), 1.98 (dd, J= 13.2, 5.9 Hz, 1H), 1.31 (s, 3H), 1.0720 0.98 (m, 1H), 0.98 - 0.87 (m, 1H); ^I9F NMR (282 MHz, DMSO-7₆) δ -58.53; MS (ES+):

588.0 (M+l), (ES-): 586.0 (M-l); Analysis calculated for C25H₂iBrF3N₇O2.1.1HC1.1.75H2O: C, 45.49; H, 3.91; Cl, 5.91; N, 14.86; Found: C, 45.32; H, 3.90; Cl, 5.87; N, 14.62.

Scheme 29

113

1. NaH.NC^CN

2. N32S2O4

NH

A

H NH₂«AcOH

Br CC^Bu

CS2CO3

29a 29b

29d 29e

29c

Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetyl)-N-(6-bromopyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (29f)

Step-1: Préparation of 2-amino-lH-pyrrolo[2,3-c]pyridine-3-carbonitrile (29b)

To NaH (2.55 g, 63.7 mmol) cooled to 0 °C was added portion-wise a solution of malononitrile (4.21 g, 63.7 mmol) in THF (40 mL). The resulting cloudy mixture was stirred at 0 °C for 1 h, followed by slow addition of 4-chloro-3-nitropyridine (29a) (5.00 g, 31.5 mmol) in THF (10 mL). The mixture was then heated at 60 °C under argon for 3 h. The cooled mixture was quenched with H2O (30 mL) and extracted with EtOAc (50 mL x 5). The 10 combined organic extract was washed with H2O (50 mL x 2), brine (50 mL), dried over anhydrous Na2SC>4, filtered and concentrated to provide 2-(3-nitropyridin-4-yl)malononitrile (8.77 g) as an orange-red solid, which was used as such in the next reaction; MS (ES+) 189 (M+l), (ES-) 187 (M-l). To 2-(3-nitropyridin-4-yI)malononitrile (4.40 g, 23.39 mmol) suspended in DMF (20 mL) at rt was added a solution NaHCCh (9.82 g, 117 mmol) in H2O 15 (20 mL) followed by solid Na2S2Û4 (12.22 g, 70.2 mmol). The resulting mixture was stirred at rt for 16 h and filtered. The filtrate was extracted with EtOAc (50 mL x 4). The combined organic extract was washed with H2O (30 mL x 4) brine (30 mL), dried, filtered and concentrated in vacuum. The residue obtained was purified by flash column chromatography (S1O2, 40 g, eluting with 0-20% DMA-80 in DCM) to provide 2-amino-lH-pyrrolo[2,320 c]pyridine-3-carbonitrile (29b) (0.673 g, 18 % yield) as a beige solid. *H NMR (300 MHz, DMSO-Jô) δ 8.30 (s, 1H), 8.03 (d, J = 5.3 Hz, 1H), 7.27 (s, 2H), 7.14 (d, J = 5.3 Hz, 1H); MS (ES+): 159 (M+l), (ES-): 157 (M-l).

Step-2: Préparation of 9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (29c)

114

To a suspension of 2-amino-lH-pyrrolo[2,3-c]pyridine-3-carbonitriIe (29b) (0.46 g, 2.91 mmol) in EtOH (10 mL) in a pressure vessel was added formamidine acetate (2.422 g, 23.27 mmol). The cloudy pale-yellow mixture was heated at 80 °C for 16 h, during which the cloudy mixture tumed to a clear solution, and then a precipitate formed. The resulting cloudy 5 pale-yellow mixture was hot-filtered. The filtered cake was washed thoroughly with boiling

EtOH to provide the product 9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (29c) (0.35 g, 65.0 % yield) as a pale-yellow solid. Ή NMR (300 MHz, DMSO-d₆) δ 12.15 (s, 1H), 8.78 (s, 1H), 8.38 (d, J = 5.3 Hz, 1H), 8.34 (s, 1H), 8.31 (d, J = 5.4 Hz, 1H), 7.49 (s, 2H); MS (ES+): 186 (M+l), (ES-): 184 (M-l).

Step-3: Préparation of tert-butyl 2-(4-amino-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetate (29d)

Compound 29d was prepared according to the procedure reported in step-1 of scheme-I, from 9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (29c) (0.35 g, 1.890 mmol) in DMF (25 mL) using tert-butyl 2-bromoacetate (0.369 g, 1.890 mmol), CS2CO3 (1.232 g, 3.78 75 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-7%] tert-butyl 2-(4amino-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (29d) (0.27 g, 48% yield) as a pale yellow solid; ‘H NMR (300 MHz, DMSO-î/₆) δ 8.94 (s, 1H), 8.45 (d, J= 5.3 Hz, 1H), 8.39 (s, 1H), 8.38 - 8.34 (m, 1H), 7.66 (s, 2H), 5.22 (s, 2H), 1.41 (s, 9H); MS (ES+): 300 20 (M+l), (ES-1): 298 (M-l).

Step-4: Préparation of 2-(4-amino-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (29e)

Compound 29e was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (29d) (270 25 mg, 0.902 mmol) in DCM (10 mL) using TFA (1029 mg, 9.02 mmol) and stirring at RT for

h. This gave after workup 2-(4-amino-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (29e) (0.53 g) TFA sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 9.51 (s, 1H), 8.96 (d, J= 6.2 Hz, 1H), 8.78 (d, J= 6.2 Hz, 1H), 8.56 (s, 1H), 5.35 (s, 2H); ¹⁹F NMR (282 MHz, DMSO-cfc) δ -74.66.; MS (ES+): 244 (M+l), (ES-): 242 (M-l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[4',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (29f)

115

Compound 29f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (29e) (50 mg, 0.140 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (70.0 mg, 0.177 mmol), HATU (63.9 mg,

0.168 mmol), DIPEA (90 mg, 0.7 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[4',3^,:4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(610 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (29f) (57 mg, 80 % yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-c/₆) δ 10.80 (s, 1H), 9.53 (s, 1H), 9.13 (d, J = 6.3 Hz, 1H), 8.94 (s, 2H), 8.80 (d, J= 6.3 Hz, 1H), 8.69 (s, 1H), 8.00 (d, J= 8.1 Hz, 1H), 7.70 (t, J= 7.9 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.96 (d, J= 17.4 Hz, 1H), 5.53 (d, J= 17.3 Hz, 1H), 4.47 - 4.44 (m, 1H), 3.91 - 3.86 (m, 1H), 2.40 - 2.29 (m, 1H), 2.29 - 2.13 (m, 1H),

2.00- 1.85 (m, 1H), 1.16-1.00 (m, 1H), 0.98 - 0.80 (m, 1H); MS (ES+): 507/509 (M+l), (ES-): 505/507 (M-l).

Scheme 30

30a

30b

HC(OMe)₃, NH₄OAc

AcOH

30c

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N20 (6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (30f)

Step-1: Préparation of 2-amino-7-methyl-lH-indole-3-carbonitrile (30b)

116

Compound 30b was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-6-methylphenyl)-2,2,2-trifluoroacetamide (30a) (7.45 g, 26.4 mmol; CAS # 2007409-96-1) in DMSO (20 mL) using malononitrile (2.094 g, 31.7 mmol), L-proline (0.608 g, 5.28 mmol), Cul (0.503 g, 2.64 mmol), a solution of K2CO3 (7.30 g, 52.80 mmol) in water (20 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [SiO₂ gel (40 g), eluting with EtOAc in hexane from 0-40%] 2amino-7-methyl-lH-indole-3-carbonitrile (30b) (2.35 g, 52% yield) as a brown solid; 'H NMR (300 MHz, DMSO-J₆) δ 10.73 (s, 1H), 6.97 (d, J= Ί.6 Hz, 1H), 6.88 (t, J= 7.5 Hz, 1H), 6.73 (d, J= 7.3 Hz, 1H), 6.49 (s, 2H), 2.33 (s, 3H); MS (ES+): 172 (M+l); (ES-): 170 (M-l).

Step-2: Préparation of 8-methyl-9H-pyrimido[4,5-b]indol-4-amine (30c)

Compound 30c was prepared according to the procedure reported in step-1 of scheme-6, from 2-aminô-7-methyl-lH-indole-3-carbonitrile (30b) (4.28 g, 25 mmol) using trimethyl orthoformate (26.5 g, 250 mmol), AcOH (4.50 g, 75 mmol) and NH4OAC (5.78 g, 75 mmol).

This gave after workup 8-methyl-9H-pyrimido[4,5-b]indol-4-amine (30c) (4.24 g, 86% yield) as a pale-yellow solid; *H NMR (300 MHz, DMSO-ifc) δ 11.81 (s, 1H), 8.26 (s, 1H), 8.10 (dd, J = 7.4, 1.8 Hz, 1H), 7.21 - 7.02 (m, 4H), 2.54 (s, 3H). MS (ES+): 199 (M+l).

Step-3: Préparation of tert-butyl 2-(4-amino-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (30d)

Compound 30d was prepared according to the procedure reported in step-1 of scheme-1, from 8-methyl-9H-pyrimido[4,5-b]indol-4-amine (30c) (3.07 g, 15.49 mmol) in DMF (20 mL) using tert-butyl 2-bromoacetate (3.02 g, 15.49 mmol) CS2CO3 (6.06 g, 18.59 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3 %] tert-butyl 2-(425 amino-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (30d) (3.52 g, 73% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-c/₆) δ 8.29 (s, 1H), 8.19 (dd, J= 5.8, 3.4 Hz, 1H), 7.26 (s, 2H), 7.17 (d, J= 2.6 Hz, 1H), 7.15 (s, 1H), 5.34 (s, 2H), 2.64 (s, 3H), 1.43 (s, 9H); MS (ES+): 313 (M+l); (ES-): 311 (M-l).

Step-4: Préparation of 2-(4-amino-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (30e)

Compound 30e was prepared according to the procedure reported in step-2 of scheme-1, from ierZ-butyl 2-(4-amino-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (30d) (0.12 g, 0.384

117 mmol) using TFA (438 mg ,3.84 mmol) in DCM (5 mL) and stirring at RT for 16 h. This gave after workup 2-(4-amino-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (30e) (0.190 g) TFA sait as a yellow solid; Ή NMR (300 MHz, DMSO-tZ₆) δ 8.87 - 8.66 (m, 2H), 8.64 (s, 1H), 8.36 (dd, J= 5.8, 3.4 Hz, 1H), 7.40 - 7.25 (m, 2H), 5.47 (s, 2H), 2.70 (s, 3H);

¹⁹F NMR (282 MHz, DMSO-J₆) δ -74.60; MS (ES+): 257 (M+l), (ES-): 255 (M-l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (30f)

Compound 30f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (30e) (50 mg,

0.135 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (43.0 mg, 0.135 mmol), HATU (61.6 mg, 0.162 mmol) DIPEA (87 mg, 0.675 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 4%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (30f) (44 mg, 63% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-îZô) δ 10.82 (s, 1H, D₂O exchangeable), 8.82 - 8.54 (m, 3H, 2H D₂O exchangeable), 8.42 - 8.28 (m, 1H), 8.00 (d, J= 8.2 Hz, 1H), 7.70 (t,

J= 8.0 Hz, 1H), 7.38 - 7.24 (m, 3H), 5.93 (d, J= 18.0 Hz, 1H), 5.65 (d, J= 17.9 Hz, 1H), 4.42 (dd, J= 9.0, 5.7 Hz, 1H), 3.96 - 3.90 (m, 1H), 2.71 (s, 3H), 2.43 - 2.28 (m, 1H), 2.25 2.12 (m, 1H), 1.99 - 1.83 (m, 1H), 1.14 - 0.99 (m, 1H), 0.74 - 0.59 (m, 1H); MS (ES+): 520.0 (M+l), (ES-): 518.0 (M-l); Analysis calculated for C₂₄H₂₂BrN7O2.1.1HC1.2.5H₂O: C, 47.60; H, 4.68; Cl, 6.44; N, 16.19; Found: C, 47.64; H, 4.48; Cl, 6.39; N, 16.14.

Scheme 31

Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (31 f)

Step-1: Préparation of 2-amino-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (31b)

NaH (1.990 g, 49.8 mmol) was added portion-wise to a cold solution of malononitrile (3.29 g, 49.8 mmol) in THF (40 mL) at 0 °C. The resulting cloudy mixture was stirred at 0 °C for 1 h, followed by slow addition of 2-bromo-3-nitropyridine (31a) (5.00 g, 24.63 mmol) in THF (10 mL). The mixture was then heated at 60 °C under argon for 3 h. The cooled mixture was quenched with H2O (30 mL) and extracted with EtOAc (50 mL x 5). The combined organic extract was washed with H2O (50 mL x 2), brine (50 mL), dried, filtered and concentrated to provide 2-(3-nitropyridin-2-yl)malononitrile (8.62 g) as an orange-red solid, which was used as such in the next reaction; MS (ES+): 189 (M+l), (ES-): 187 (M-l). A suspension of 2-(3nitropyridin-2-yl)malononitrile (4.315 g, 22.93 mmol) zinc (7.50 g, 115 mmol) in acetic acid (27.5 g, 459 mmol) was heated at 60 °C for 2 h and filtered hot. The filtered cake was washed thoroughly with boiling EtOH. The filtrate was concentrated to dryness. The concentrate was suspended in H2O (50 mL), neutralized with 3 M aqueous NaOH until pH 7, and then extracted with EtOAc (50 mL x 3). The combined extract was washed with H2O (30 mL x 2), brine (30 mL), dried, filtered and concentrated in vacuum. The residue obtained was purified by flash column chromatography (S1O2, 40 g, eluting with 0-20% DMA-80 in DCM) to provide 2-amino-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (31b) (320 mg, 9 % yield) as beige solid; Ή NMR (300 MHz, DMSO-_d6) δ 10.82 (s, 1H), 8.04 (dd, J = 5.0, 1.4 Hz, 1H), 7.36

119 (dd, J = 7.8, 1.4 Hz, 1H), 7.16 (s, 2H), 6.86 (dd, J = 7.8, 4.9 Hz, 1H); MS (ES+): 159 (M+l), (ES-): 157 (M-l).

Step-2: Préparation of 9H-pyrido[2',3^,:4,5]pyrrolo[2,3-d]pyrimidin-4-amine (31c)

Compound 31c was prepared according to the procedure reported in step-1 of scheme-6, from 5 2-amino-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (31b) (0.31 g, 1.960 mmol) using trimethyl orthoformate (4.16 g, 39.2 mmol), AcOH (0.589 g, 9.80 mmol) andNHziOAc (0.755 g, 9.80 mmol). This gave after workup and purification [silica gel (12 g), eluting with DMA-80 in DCM from 0-30%] 9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (31c) (100 mg, 28% yield) as a pale-yellow solid; *H NMR (300 MHz, DMSO-Je) δ 12.02 (d, J = 2.0 Hz, 1H), 10 8.49 (dd, J = 4.8, 1.4 Hz, 1H), 8.34 (s, 1H), 7.83 (dd, J = 8.1, 1.4 Hz, 1H), 7.37 (dd, J = 8.2,

4.8 Hz, 1H); MS (ES+): 186 (M+l), (ES-): 184 (M-l).

Step-3: Préparation of /eri-butyl 2-(4-amino-9H-pyrido[2',3^,:4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetate (31d)

Compound 31d was prepared according to the procedure reported in step-2 of scheme-16, 15 9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (31c) (80 mg, 0.432 mmol) in DMF (2.5 mL) using fërf-butyl 2-bromoacetate (0.077 mL, 0.518 mmol), CS2CO3 (282 mg, 0.864 mmol) and stirring at RT for 1.5 h under nitrogen atmosphère and quenching by adding water. The solid separated was filtered and dried to give tert-butyl 2-(4-amino-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (31d) (109 mg, 84 % yield) as a pale 20 yellow solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.54 (dd, J= 4.9, 1.3 Hz, 1H), 8.39 (s, 1H),

8.04 (dd, J= 8.3, 1.3 Hz, 1H), 7.43 (dd, J= 8.2, 4.8 Hz, 1H), 5.17 (s, 2H), 1.40 (s, 9H); MS (ES+): 300.1 (M+l).

Step-4: Préparation of 2-(4-amino-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (31e)

Compound 31e was prepared according to the procedure reported in step-2 of scheme-1, from terributyl 2-(4-amino-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (31d) (80 mg, 0.267 mmol) using 20% TFA in DCM (1534 pL, 4.01 mmol) and stirring at RT for 16 h.

This gave after workup 2-(4-amino-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (31e) (64 mg, 98% yield) TFA sait as a yellow solid; *H NMR (300 MHz, DMSO-dô) δ 30 8.65 (dd, J= 4.9, 1.3 Hz, 1H), 8.55 (s, 1H), 8.25 (d, J= 8.3 Hz, 1H), 7.56 (dd, J= 8.3, 4.9

Hz, 1H), 5.28 (s, 2H); MS (ES+): 244.10 (M+l).

120

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (310 .

Compound 31f was prepared according to the procedure reported in step-3 of scheme-1, from

TFA sait of 2-(4-amino-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (31e) (60 mg, 0.247 mmol) in DMF (2 mL) using TFA sait of (lR,3S,5R)-N-(6-bromopyridin-2yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (98.0 mg, 0.247 mmol), HATU (141 mg, 0.370 mmol), DIPEA (0.215 mL, 1.233 mmol) and stirring at RT for 1 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with .

DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (31f) (79 mg, 63% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/₆) δ 10.77 (s, 1H, D₂O exchangeable),

8.97 (s, 1H, D₂O exchangeable), 8.75 - 8.64 (m, 2H), 8.31 (d, J= 8.4 Hz, 1H), 8.00 (d, J =

8.2 Hz, 1H), 7.75 - 7.61 (m, 2H), 7.31 (d, J= 7.7 Hz, 1H), 5.84 (d, J= 17.4 Hz, 1H), 5.47 (d, J= 17.3 Hz, 1H), 4.45 - 4.37 (m, 1H), 3.89 (ddd, J= 7.5, 5.4, 2.4 Hz, 1H), 2.40 - 2.13 (m, 2H), 1.99 - 1.82 (m, 1H), 1.07 (dt, J= 8.7, 5.4 Hz, 1H), 0.80 (td, J= 5.2, 2.3 Hz, 1H); MS (ES+): 507.1 (M+l); (ES-): 505.0 (M-l); Analysis calculated for C₂₂Hi₉BrN₈O₂

1.75H₂O.1.2HC1: C, 45.35; H, 4.10; Cl, 7.30; N, 19.23; Found: C, 45.38; H, 4.02; Cl, 7.16;

N, 18.95.

Scheme 32

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (32f)

Step-1: Préparation of 2-amino-5-fluoro-lH-indole-3-carbonitrile (32b)

Compound 32b was prepared according to the procedure reported in step-1 of scheme-11, from2,2,2-trifluoro-N-(4-fluoro-2-iodophenyl)acetamide (32a) (7.03 g, 21.1 mmol; CAS # 784183-55-7) in DMSO (30 mL) using malononitrile (1.673 g, 25.3 mmol), L-proline (0.486 g, 4.22 mmol), Cul (0.402 g, 2.110 mmol), K2CO3 (5.83 g, 42.2 mmol) and heating at 60 °C for 15 h under an argon atmosphère. This gave after workup and purification [S1O2 gel (80

g), eluting with EtOAc in hexane from 0-50%] 2-amino-5-fluoro-lH-indole-3-carbonitrile (32b) (2.657 g, 72% yield) as a brown solid; MS (ES+): 176.05 (M+l).

Step-2: Préparation of 6-fluoro-9H-pyrimido[4,5-b]indol-4-amine (32c)

Compound 32c was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5-fluoro-lH-indole-3-carbonitrile (32b) (1.2 g, 6.85 mmol) in éthanol (30 mL) using formamidine acetate (5.76 g, 54.8 mmol) and refluxing for 40 h. This gave after workup 6-fluoro-9H-pyrimido[4,5-b]indol-4-amine (32c) as a brown solid (2.706 g) which was used as such in the next step; MS (ES+): 203.00 (M+l).

Step-3: Préparation of iert-butyl 2-(4-amino-6-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (32d)

122

Compound 32d was prepared according to the procedure reported in step-1 of scheme-1, from 6-fluoro-9H-pyrimido[4,5-b]indol-4-amine (32c) (346 mg, 1.71 mmol) in DMF (10 mL) using tert-butyl 2-bromoacetate (0.252 mL, 1.71 mmol), césium carbonate (1.337 g) and stirring at RT for 47 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with hexanes/10% methanol in ethyl acetate (1:0 to 1:1)] tert-butyl 2-(4-amino-6-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (32d) (160 mg, 30% yield) as a light yellow solid; 1H NMR (300 MHz, DMSO-J₆) δ 8.32 - 8.22 (m, 2H), 7.57 (dd, J= 8.9, 4.5 Hz, 1H), 7.38 (s, 2H), 7.26 (td, J= 9.2, 2.5 Hz, 1H), 5.12 (s, 2H), 1.39 (s, 9H); ¹⁹F NMR (282 MHz, DMSO-cfe) δ -122.42; MS (ES+): 317.10 (M+l).

Step-4: Préparation of 2-(4-amino-6-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (32e)

Compound 32e was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-6-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (32d) (140 mg, 0.443 mmol) in DCM (10 mL) using TFA. The reaction mixture was concentrated to dryness to afford 2-(4-amino-6-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (32e) which used as /5 such for the next step; MS (ES+): 261.10 (M+l); (ES-): 259.00 (M-l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-fluoro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (32f)

Compound 32f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (32e) (0.443 mmol, from above step-4) in DMF (15 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (141.0 mg, 0.443 mmol), HATU (337 mg, 0.886 mmol) DIPEA (0.386 mL, 2.215 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by treating of product obtained with acetonitrile (2 mL) and 50 mM aq. HCl (8 mL) and lyophilization (lR,3S,5R)-2-(2-(4amino-6-fluoro-9H-pyrimido [4,5 -b] indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (32f) (53 mg, 23% yield) HCl sait as a light brown solid; Ή NMR (300 MHz, DMSO-<7₆) δ 10.76 (s, 1H, D₂O exchangeable), 8.73 (s, 2H, D₂O exchangeable), 8.63 (s, 1H), 8.47 (dd, J= 9.8, 2.5 Hz, 1H), 8.00 (d, J= 8.1 Hz, 1H), 7.79 30 7.65 (m, 2H), 7.45 (td, J= 9.2, 2.5 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.78 (d, J= 17.4 Hz,

1H), 5.43 (d, J= 17.3 Hz, 1H), 4.41 (dd, J= 9.0, 5.5 Hz, 1H), 4.02 - 3.82 (m, 1H), 2.41 2.12 (m, 2H), 1.99-1.79 (m, 1H), 1.16-0.97 (m, 1H), 0.82-0.71 (m, 1H); ¹⁹FNMR(282

123

MHz, DMSCM;) δ -119.89; MS (ES+): 524.10 & 526.10 (M+l); MS (ES-): 522.00 & 524.00 (M-l); Analysis calculated for C₂3Hi₉BrFN₇O2.1.2HC1.2.75H2O: C, 44.73; H, 4.19; N, 15.87; Cl, 6.89; Found: C, 44.84; H, 4.13; N, 15.51; Cl, 6.81.

Scheme 33

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (33e)

Step-1: Préparation of 7-fluoro-9H-pyrimido[4,5-b]indol-4-amine (33b)

Compound 33b was prepared according to the procedure reported in step-2 of scheme-29, 10 from 2-amino-6-fluoro-lH-indole-3-carbonitrile (33a) (1.2 g, 6.85 mmol; CAS # 378236-807) in éthanol (30 mL) using formamidine acetate (5.76 g, 54.8 mmol) and refluxing for 20 h. This gave after workup 7-fluoro-9H-pyrimido[4,5-b]indol-4-amine (33b) as a brown solid (2.62 g) which was used as such in the next step; *H NMR (300 MHz, DMSO-îZô) δ 8.29 (dd, J= 8.7, 5.4 Hz, 1H), 8.23 (s, 1H), 7.78 (s, 1H), 7.22 (dd, J= 9.Ί, 2.4 Hz, 1H), 7.10 - 7.01 (m,

1H); ¹⁹F NMR (282 MHz, DMSO-<7₆) δ -116.72; MS (ES+): 203.10 (M+l).

Step-2: Préparation of terributyl 2-(4-amino-7-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (33c)

Compound 33c was prepared according to the procedure reported in step-1 of scheme-1, from 7-fluoro-9H-pyrimido[4,5-b]indol-4-amine (33b) (346 mg, 1.71 mmol) in DMF (10 mL)

124 using ZerZ-butyl 2-bromoacetate (0.252 mL, 1.71 mmol), césium carbonate (1.337 g) and stirring at RT for 46 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with hexanes/10% methanol in ethyl acetate (1:0 to 1:1)] ZerZ-butyl 2-(4-amino-7-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (33c) (151 mg,

28% yield) as a light yellow solid; 1H NMR (300 MHz, DMSO-î/6) δ 8.35 (dd, J= 8.7, 5.4

Hz, 1H), 8.28 (s, 1H), 7.53 (dd, J= 10.2, 2.4 Hz, 1H), 7.31 (s, 2H), 7.17-7.04 (m, 1H), 5.11 (s, 2H), 1.40 (s, 9H); ¹⁹F NMR (282 MHz, DMSO-ri₆) δ -116.10; MS (ES+): 317.20 (M+l). Step-3: Préparation of 2-(4-amino-7-fhioro-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (33d) Compound 33d was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-7-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (33c) (145 mg, 0.458 mmol) in DCM (10 mL) using TFA. This gave after workup 2-(4-amino-7-fluoro-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (33d) which was used as such in next step; MS (ES+): 261.10 (M+l); (ES-): 259.00 (M-l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-7-fluoro-9H-pyrimido[4,5-b]indol-915 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (33e)

Compound 33e was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (33d) (119 mg, 0.458 mmol) in DMF (15 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (146.0 mg, 0.458 mmol), HATU (348 mg,

0.916 mmol), DIPEA (0.399 mL, 2.29 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [Cl 8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] followed by purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 5%], dissolving the product in acetonitrile (3 mL) and 0.1% aq. HCl (20 mL) followed by lyophilization (lR,3S,5R)-2-(2-(4-amino-7-fluoro-9H-pyrimido[4,5-b]indol-9-yI)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (33e) (31 mg, 13% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-ri₆) δ 10.77 (s, 1H, D₂O exchangeable), 8.76 - 8.66 (m, 3H, D₂O exchangeable), 8.63 (s, 1H), 8.56 (dd, J= 8.8, 5.2 Hz, 1H), 8.00 (d, J= 8.2 Hz, 1H), 7.75 - 7.63 (m, 2H), 7.37 - 7.25 (m, 2H), 5.76 (d, J= 17.4 Hz, 1H), 5.40 (d,

J= 17.3 Hz, 1H), 4.42 (dd, J=9.1, 5.5 Hz, 1H), 3.95-3.84 (m, 1H), 2.42-2.12 (m, 2H), 2.02 - 1.76 (m, 1H), 1.12 - 0.99 (m, 1H), 0.89 - 0.75 (m, 1H); ¹⁹F NMR (282 MHz, DMSOd₆) δ -113.65; MS (ES+): 524.10 & 526.10 (M+l); MS (ES-): 522.10 & 524.00 (M-l);

125

Analysis calculated for C23Hi9BrFN₇O2.1.1HC1.2.0H₂O: C, 46.00; H, 4.05; Cl, 6.49; N, 16.33; Found: C, 46.28; H, 4.10; Cl, 6.44; N, 15.96.

Scheme 34

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (34e)

Step-1: Préparation of 6-bromo-9H-pyrimido[4,5-b]indol-4-amine (34b)

Compound 34b was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-5-bromo-lH-indole-3-carbonitrile (34a) (2.20 g, 9.32 mmol; CAS # 124214010 64-2) using triethyl orthoformate (31.0 mL, 186 mmol), AcOH (2.66 mL, 46.6 mmol) and

NH4OAC (3.59 g, 46.6 mmol). This gave after work up 6-bromo-9H-pyrimido[4,5-b]indol-4amine (34b) (1.58 g, 64% yield) as a pale yellow solid; 'H NMR (300 MHz, DMSO-cfc) δ 11.99 (s, 1H), 8.58 (d, J= 1.9 Hz, 1H), 8.26 (s, 1H), 7.48 (dd, J= 8.6, 1.8 Hz, 1H), 7.39 (d, J = 8.6 Hz, 1H), 7.30 (s, 2H); MS (ES+): 263/265 (M+l).

Step-2: Préparation of tert-butyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (34c)

Compound 34c was prepared according to the procedure reported in step-1 of scheme-1, from 6-bromo-9H-pyrimido[4,5-b]indol-4-amine (34b) (1.58 g, 6.01 mmol) in DMF (20 mL) using tert-butyl 2-bromoacetate (1.171 g, 6.01 mmol), Cs₂CC>3 (2.348 g, 7.21 mmol) and stirring at

126

RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] ZerZ-butyl 2-(4-amino-6-bromo-9Hpyrimido[4,5-b]indol-9-yl)acetate (34c) (2.265 g, 72% yield) as a pale orange solid; 'H NMR (300 MHz, DMSO-ί/ό) δ 8.64 (s, 1H), 8.31 (s, 1H), 7.56 (s, 2H), 7.44 (s, 2H), 5.13 (s, 2H), 5 1.40 (s, 9H); MS (ES+): 377/379 (M+l), (ES-): 375/377 (M-l).

Step-3: Préparation of 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (34d)

Compound 34d was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indoI-9-yl)acetate (34c) (0.300 g, 0.795 mmol) in DCM (10 mL) using TFA (0.907 g, 7.95 mmol) and stirring at RT for 16 h.

This gave after workup 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (34d) (0.387 g) TFA sait as a yellow solid; ’H NMR (300 MHz, DMSO-î/₆) δ 8.79 (s, 1H), 8.70 8.44 (m, 3H), 7.77 (d, J= 8.8 Hz, 1H), 7.69 (dd, J= 8.7, 1.8 Hz, 1H), 5.26 (s, 2H); MS (ES+): 321/323 (M+l), (ES-): 319/321 (M-l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-915 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (34e)

Compound 34e was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (34d) (50 mg, 0.115 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (36.6 mg, 0.115 mmol), HATU (52.4 mg, 0.138 mmol) DIPEA (74.3 mg, 0.575 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl 8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(625 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (34e) (26 mg, 39% yield) HCl sait as a white solid; ’HNMR (300 MHz, DMSO-cfe) δ 10.75 (s, 1H, D₂O exchangeable), 8.76 (d, J= 3.1 Hz, 1H), 8.54 (d, J= 5.2 Hz, 1H), 8.38 (s, 2H, D₂O exchangeable), 8.00 (d, J= 8.1 Hz, 1H), 7.75-7.58 (m, 3H), 7.32 (d,J=7.7Hz, 1H), 5.73 (dd, J= 17.7, 1.8 Hz, 1H), 5.39 (d, J= 17.4 Hz, 1H), 4.40 (dd, J- 9.2, 5.5 Hz, 1H), 3.94 - 3.86 (m, 1H), 2.38 - 2.26 (m, 1H),

2.26-2.15 (m, 1H), 1.96-1.84 (m, 1H), 1.13-0.97 (m, 1H), 0.81-0.72 (m, 1H); MS (ES+) 584.0 (M+l); (ES-): 582.0 (M-l); Analysis calculated for C₂₃Hi9Br₂N₇O₂.HC1.2.25H₂O: C, 41.71; H, 3.73; Cl, 5.35; N, 14.81; Found: C, 41.55; H, 3.58; Cl, 5.49; N, 14.65.

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (35f)

Step-1 : Préparation of 2-amino-7-bromo-lH-indole-3-carbonitrile (35b)

Compound 35b was prepared according to the procedure reported in step-1 of scheme-11, from N-(2,6-dibromophenyl)-2,2,2-trifluoroacetamide (35a) (7.79 g, 22.45 mmol; CAS # '

340034-49-3) in DMSO (20 mL) using malononitrile (1.780 g, 26.9 mmol), L-proline (0.517 g, 4.49 mmol), Cul (0.428 g, 2.245 mmol), K2CO3 (6.21 g, 44.9 mmol) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [S1O2 gel (40 g), eluting with EtOAc in hexane from 0-40%] 2-amino-7-bromo-lH-indole-3-carbonitrile (35b) (4.18 g, 18% yield) as a pale orange solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.91 (s, 1H), 7.15 (dd, J = 7.7, 1.0 Hz, 1H), 7.11 (dd, J = 7.9, 0.9 Hz, 1H), 6.93 (t, J = 7.8 Hz, 1H), 6.69 (s, 2H); MS (ES+): 236 ; (ES-): 234 (M-l).

Step-2: Préparation of 8-bromo-9H-pyrimido[4,5-b]indol-4-amine (35c)

Compound 35c was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-7-bromo-lH-indole-3-carbonitrile (35b) (4.18 g, 17.71 mmol) using trimethyl

128 orthoformate (37.6 mg, 354 mmol), AcOH (5.06 mL, 89 mmol) and NFLOAc (6.82 g, 89 mmol). This gave after workup 8-bromo-9H-pyrimido[4,5-b]indol-4-amine (35c) as a paleyellow solid residue was used as such for the next step; 'H NMR (300 MHz, DMSO-i/e) δ 11.97 (s, 1H), 8.32 (d, J= 7.9 Hz, 2H), 7.56 (d, J= 7.8 Hz, 1H), 7.30 (s, 2H), 7.17 (t, J= 7.8 Hz, 1H).

Step-3: Préparation of tert-butyl 2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (35d)

Compound 35d was prepared according to the procedure reported in step-1 of scheme-1, from 8-bromo-9H-pyrimido[4,5-b]indol-4-amine (35c) (4.11 g, 15.62 mmol) in DMF (20 mL) using terributyl 2-bromoacetate (3.05 g, 15.62 mmol) CS2CO3 (6.62 g, 20.3 Immol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3 %] /eri-butyl 2-(4amino-8-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (35d) (4.11 g, 88% yield) as a pale orange solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.39 (d, J= 7.8 Hz, 1H), 8.34 (s, 1H), 7.59 (d, J= 7.8 Hz, 1H), 7.48 (s, 2H), 7.21 (t, J= 7.8 Hz, 1H), 5.44 (s, 2H), 1.43 (s, 9H).

Step-4: Préparation of 2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (35e)

Compound 35e was prepared according to the procedure reported in step-2 of scheme-1, from fërributyl 2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (35d) (0.250 g, 0.663 mmol) using TFA (1.511 g, 13.25 mmol) in DCM (20 mL) and stirring at RT for 16 h. This gave after workup 2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (35e) (0.356 g) TFA sait as a yellow solid; Ή NMR (300 MHz, DMSO-cfc) δ 8.57 (s, 1H), 8.51 (d, J= 7.9 Hz, 1H), 8.49 - 8.24 (m, 2H), 7.72 (d, J= 7.8 Hz, 1H), 7.32 (t, J= 7.9 Hz, 1H), 5.54 (s, 2H).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (35f)

Compound 35f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indol-9-yI)acetic acid (35e) (50 mg, 0.115 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (36.6 mg, 0.115 mmol), HATU (52.4 mg, 0.138 mmol), DIPEA (74.3 mg, 0.575 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography

129

[C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-bromo-9H-pynmido[4,5-b]indoI-9-yI)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (35f) (38 mg, 57%yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/ô) δ 10.76 (s, 1H, D₂O exchangeable), 8.59 (s, 1H), 8.55 - 8.32 (m, 3H, 2H D₂O exchangeable), 8.00 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 7.4 Hz, 2H), 7.30 (dt, J= 7.9, 4.0 Hz, 2H), 5.98 (d, J= 17.7 Hz, 1H), 5.84 - 5.72 (m, 1H), 4.41 (dd, J= 9.0, 5.7 Hz, 1H), 3.91 - 3.87 (m, 1H), 2.40 - 2.25 (m, 1H), 2.25 - 2.15 (m, 1H), 2.00 - 1.84 (m, 1H), 1.12 - 0.99 (m, 1H), 0.81 - 0.69 (m, 1H); MS (ES+): 584.0 (M+l), (ES-): 582.0 (M-l); Analysis calculated for C₂₃Hi₉Br₂N₇O₂.HC1.2H₂O: C, 42.00; H, 3.68; Cl, 5.39; N, 14.91; Found: C, 41.98; H, 3.61; Cl, 5.08; N, 14.77.

Scheme 36

36a

36d

36b ^36c

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (36e)

Step-1: Préparation of 7-methoxy-9H-pyrimido[4,5-b]indol-4-amine (36b)

Compound 36b was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-6-methoxy-lH-indole-3-carbonitrile (36a) (2.50 g, 13.35 mmol; CAS # 1016680-93-5) using triethyl orthoformate (44.4 mL, 267 mmol), AcOH (3.82 mL, 66.8 mmol) and NH4OAC (5.15 g, 66.8 mmol). This gave after work up 7-methoxy-9Hpyrimido[4,5-b]indol-4-amine (36b) (0.89 g, 31% yield) as a brown yellow solid; *H NMR (300 MHz, DMSO-Jô) δ 11.70 (s, 1H), 8.17 (d, J= 9.9 Hz, 2H), 7.01 (s, 2H), 6.93 (d, J= 2.3 Hz, 1H), 6.82 (dd, 1H), 3.83 (s, 3H).

130

Step-2: Préparation of /er/-butyl 2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetate (36c)

Compound 36c was prepared according to the procedure reported in step-1 of scheme-1, from 7-methoxy-9H-pyrimido[4,5-b]indol-4-amine (36b) (0.89 g, 4.15 mmol) in DMF (20 mL) using Zert-butyl 2-bromoacetate (0.81 g, 4.15 mmol) and CS2CO3 (1.624 g, 4.99 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] fëri-butyl 2-(4amino-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (36c) (0.83 g, 61% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-7₆) δ 8.22 (d, J= 7.5 Hz, 2H), 7.22 - 7.07 (m, 3H),

6.89 (dd, J= 8.6, 2.3 Hz, 1H), 5.10 (s, 2H), 3.85 (s, 3H), 1.41 (s, 9H).

Step-3: Préparation of 2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (36d)

Compound 36d was prepared according to the procedure reported in step-2 of scheme-1, from terZ-butyl 2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (36c) (0.83 g, 2.53 mmol) in DMF (20 mL) using TFA (5.76 g, 50.6 mmol) and stirring at RT for 16 h. The reaction mixture was concentrated in vacuum to afford 2-(4-amino-7-methoxy-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (36d) (1.18 g) TFA sait as a yellow solid; *H NMR (300 MHz, DMSO-7₆) δ 8.61 (s, 2H), 8.59 (s, 1H), 8.41 (d, J= 8.8 Hz, 1H), 7.46 (d, J= 2.3 Hz, 1H), 7.07 (dd, J= 8.8, 2.2 Hz, 1H), 5.28 (s, 2H), 3.89 (s, 3H).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-920 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (36e)

Compound 36e was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (36d) (50 mg, 0.129 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (41.2 mg, 0.129 mmol), HATU (59.1 mg, 0.155 mmol), DIPEA (84 mg, 0.647 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(630 bromopyridin-2-yl)-2-azabicycIo[3.1.0]hexane-3-carboxamide (36e) (52 mg, 75% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-ifc) δ 10.67 (s, 1H, D2O exchangeable), 8.48 (s, 1H), 8.35 (dd, J = 12.4, 6.2 Hz, 3H, 2H D₂O exchangeable), 7.94 (d, J = 8.2 Hz, 1H),

131

7.64 (t, J = 8.0 Hz, 1H), 7.25 (d, J = 7.7 Hz, 1H), 7.19 (d, J = 2.2 Hz, 1H), 6.97 (dd, J = 8.7, 2.2 Hz, 1H), 5.64 (d, J = 17.3 Hz, 1H), 5.35 (d, J = 17.2 Hz, 1H), 4.40 - 4.29 (m, 1H), 3.88 3.80 (m, 4H), 2.34-2.23 (m, 1H), 2.23-2.10 (m, 1H), 1.92-1.80 (m, 1H), 1.06-0.95 (m, 1H), 0.74 - 0.64 (m, 1H); MS (ES+): 536.0 (M+l), (ES-): 534.0 (M-l); Analysis calculated for C24H₂2BrN₇O3.1.2HC1.2.25H2O: C, 46.44; H, 4.50; Cl, 6.85; N, 15.80; Found: C, 46.47; H, 4.50; Cl, 6.46; N, 15.867.

Scheme 37

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indoI-9-yl)acetyl)10 N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (37a)

Compound 37a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (36d) (100 mg, 0.259 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (86 mg, 0.259 mmol), HATU (118 mg, 0.311 mmol), DIPEA (167 mg, 1.294 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)20 N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (37a) (85 mg, 60% yield) HCI sait as a white solid; 'H NMR (300 MHz, DMSO-7₆) δ 10.73 (s, 1H, D₂O exchangeable), 8.55 (s, 1H), 8.46 (s, 2H, D2O exchangeable), 8.39 (d, J= 8.8 Hz, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 7.26 (d, J= 2.3 Hz, 1H), 7.04 (dd, J= 8.7, 2.3 Hz, 1H), 5.67 (d, J= 17.3 Hz, 1H), 5.37 (d, J= 17.3 Hz, 1H), 4.37 (dd, J = 9.2, 5.9 Hz, 1H), 3.87 (s, 3H), 3.69 (dd, J= 5.5, 2.3 Hz, 1H), 2.54-2.47 (m, 1H),

132

1.99 (dd, J= 13.2, 5.9 Hz, 1H), 1.31 (s, 3H), 1.07-0.97 (m, 1H), 0.96-0.83 (m, 1H); MS (ES+): 550.0 (M+l); (ES-): 548.0 (M-l).

Scheme 38

38a 38b 38c

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (38e)

Step-1 : Préparation of 6-methoxy-9H-pyrimido[4,5-b]indol-4-amine (38b)

Compound 38b was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-5-methoxy-lH-indole-3-carbonitrile (38a) (3.10 g, 16.56 mmol; CAS #

1304143-87-0) using trimethyl orthoformate (35.1 g, 331 mmol), AcOH (4.74 mL, 83 mmol) and NH4OAC (6.38 g, 83 mmol). This gave after work up 6-methoxy-9H-pyrimido[4,5b]indol-4-amine (38b) (2.78 g, 78% yield) as a pale yellow solid; 'H NMR (300 MHz, DMSO-îZô) δ 11.62 (s, 1H), 8.20 (s, 1H), 7.85 (d, J= 2.4 Hz, 1H), 7.32 (d, J= 8.7 Hz, 1H), 7.17 (s, 2H), 6.96 (dd, J= 8.7, 2.4 Hz, 1H), 3.85 (s, 3H). MS (ES+): 215 (M+l), (ES-): 213 (M-l).

Step-2: Préparation of tert-butyl 2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetate (38c)

Compound 38c was prepared according to the procedure reported in step-1 of scheme-1, from 6-methoxy-9H-pyrimido[4,5-b]indol-4-amine (38b) (0.59 g, 2.75 mmol) in DMF (25 mL) using terributyl 2-bromoacetate (0.537 g, 2.75 mmol) CS2CO3 (1.077 g, 3.30 mmol) and

133 stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] ZerZ-butyl 2-(4amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (38c) (0.56 g, 62% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-cfc) δ 8.25 (s, 1H), 7.91 (d, J= 2.4 Hz, 1H), 7.45 (d, J= 8.8 Hz, 1H), 7.31 (s, 2H), 7.02 (dd, J= 8.8, 2.4 Hz, 1H), 5.07 (s, 2H), 3.87 (s, 3H),

1.40 (s, 9H).; MS (ES+): 329 (M+l), (ES-): 327 (M-l).

Step-3: Préparation of 2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (38d)

Compound 38d was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (38c) (0.56 g,

1.705 mmol) in DCM (20 mL) using TFA (1.945 g, 17.05 mmol) and stirring at RT for 16 h.

This gave after workup 2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (38d) (0.68 g,) TFA sait as a yellow solid; ’H NMR (300 MHz, DMSO-<7₆) δ 8.69 - 8.44 (m, 3H), 8.05 (d, J= 2.4 Hz, 1H), 7.70 (d, J= 9.0 Hz, 1H), 7.17 (dd, J= 8.9, 2.4 Hz, 1H), 5.23 (s, 2H), 3.89 (s, 3H). ¹⁹F NMR (282 MHz, DMSO-ifc) δ -74.16; MS (ES+): 273 (M+l), (ES-):

271 (M-l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (38e)

Compound 38e was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (38d) (50 mg, 20 0.129 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (41.2 mg, 0.0.129 mmol), HATU (59.1 mg, 0.155 mmol), DIPEA (84 mg, 0.647 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl 8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (38e) (41 mg, 59% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 10.75 (s, 1H), 8.81 (s, 2H), 8.60 (s, 1H), 8.07 (s, 1H), 7.99 (d, J= 8.1 Hz, 1H), 7.69 (t, J= 8.0 Hz, 1H), 7.61 (d, J= 9.0

Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 7.18 (d, J= 8.9 Hz, 1H), 5.71 (d, J= 17.3 Hz, 1H), 5.39 (d, J= 17.2 Hz, 1H), 4.40 (dd, J= 9.0, 5.5 Hz, 1H), 3.90 - 3.86 (m, 4H), 2.40 - 2.27 (m, 1H), 2.27-2.13 (m, 1H), 1.98- 1.83 (m, 1H), 1.12-1.00 (m, 1H), 0.84-0.70 (m, 1H); MS (ES+): 536/538 (M+l), (ES-): 534/536 (M-l); Analysis calculated for

134

C24H22BrN₇O3.HC1.2.75H₂O: C, 46.31; H, 4.62; Cl, 5.70; N, 15.75; Found: C, 46.48; H, 4.40;

Cl, 5.45; N, 15.18.

Scheme 39

39f

Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxylate (39f)

Step-1: Préparation of methyl 2-amino-3-cyano-lH-indole-5-carboxylate (39b)

Compound 39b was prepared according to the procedure reported in step-1 of scheme-11, from methyl 3-iodo-4-(2,2,2-trifluoroacetamido)benzoate (39a) (6.61 g, 17.72 mmol; CAS #

848485-43-8) in DMSO (20 mL) using malononitrile (1.405 g, 21.26 mmol), L-proline (0.408 g, 3.54 mmol), Cul (0.337 g, 1.772 mmol), K2CO3 (4.90 g, 35.4 mmol) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [S1O2 gel (40 g), eluting with EtOAc in hexane from 0-40%] methyl 2-amino-3-cyano-lH-indole-5carboxylate (39b) (2.94 g, 77% yield) as a brown solid; 'H NMR (300 MHz, DMSO-dô) δ

11.12 (s, 1H), 7.73 (d, J= 1.6 Hz, 1H), 7.58 (dd, J= 8.2, 1.7 Hz, 1H), 7.22 (d, J= 8.3 Hz,

1H), 7.06 (s, 2H), 3.83 (s, 3H); MS (ES+): 216, (ES-): 214 (M-l).

135

Step-2: Préparation of methyl 4-amino-9H-pyrimido[4,5-b]indole-6-carboxylate (39c)

Compound 39c was prepared according to the procedure reported in step-1 of scheme-6, from methyl 2-amino-3-cyano-lH-indole-5-carboxylate (39b) (2.94 g, 13.66 mmol) using trimethyl orthoformate (29.0 g, 273 mmol), AcOH (3.91 mL, 68.3 mmol) andNFLOAc (5.27 5 g, 68.3 mmol). The pale-yellow solid residue obtained after workup was used as such for the next step; Ή NMR (300 MHz, DMSO-î/₆) δ 12.25 (s, 1H), 8.95 (s, 1H), 8.30 (s, 1H), 7.99 (dd, J= 8.5, 1.6 Hz, 1H), 7.52 (d, J= 8.5 Hz, 1H), 7.42 (s, 2H), 3.89 (s, 3H).

Step-3: Préparation of methyl 4-amino-9-(2-(terZ-butoxy)-2-oxoethyl)-9H-pyrimido[4,5b]indole-6-carboxylate (39d)

Compound 39d was prepared according to the procedure reported in step-1 of scheme-1, from methyl 4-amino-9H-pyrimido[4,5-b]indole-6-carboxylate (39c) (2.68 g, 11.06 mmol) in DMF (20 mL) using tert-butyl 2-bromoacetate (2.158 g, 11.06 mmol), CS2CO3 (4.33 g, 13.28 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3 %] methyl 4-amino15 9-(2-(/eri-butoxy)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxylate (39d) (2.50 g, 63% yield) as a pale orange solid; ’H NMR (300 MHz, DMSO-<7₆) δ 9.00 (d, J= 1.6 Hz, 1H), 8.34 (s, 1H), 8.04 (dd, J= 8.6, 1.6 Hz, 1H), 7.69 (d, J= 8.6 Hz, 1H), 7.54 (s, 2H), 5.18 (s, 2H), 3.91 (s, 3H), 1.40 (s, 9H).

Step-4: Préparation of 2-(4-amino-6-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (39e)

Compound 39e was prepared according to the procedure reported in step-2 of scheme-1, from methyl 4-amino-9-(2-(terZ-butoxy)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxylate (39d) (2.50 g, 7.02 mmol) using TFA (16 g, 140 mmol) in DCM (20 mL) and stirring at RT for 16 h. This gave after workup 2-(4-amino-6-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol25 9-yl)acetic acid (39e) (3.70 g) TFA sait as a yellow solid; ’H NMR (300 MHz, DMSO-îZô) δ 9.16 (d, J= 1.6 Hz, 1H), 8.65 (s, 2H), 8.60 (s, 1H), 8.13 (dd, J= 8.6, 1.6 Hz, 1H), 7.89 (d, 8.7 Hz, 1H), 5.31 (s, 2H), 3.93 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-J₆) δ -74.65.

Step-5: Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-630 carboxylate (39f)

Compound 39f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (39e)

136

(50 mg, 0.121 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (38.5 mg, 0.121 mmol), HATU (55.1 mg, 0.145 mmol), DIPEA (78 mg, 0.603 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with

MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyI)-9H-pyrimido[4,5-b]indole-6-carboxylate (39f) (53 mg, 78% yield) HCl sait as a white solid; ^!H NMR (300 MHz, DMSO-î7₆) δ 10.76 (s, 1H,

D₂O exchangeable), 9.13 (s, 1H), 8.59 (s, 1H), 8.54 (s, 2H, D₂O exchangeable), 8.12 (dd, J= 8.7, 1.6 Hz, 1H), 8.00 (d, J= 8.1 Hz, 1H), 7.77 (d, J= 8.8 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.79 (d, J= 17.4 Hz, 1H), 5.44 (d, J= 17.4 Hz, 1H), 4.41 (dd, J= 8.9, 5.5 Hz, 1H), 3.92 (s, 4H), 2.38-2.28 (m, 1H), 2.28-2.14 (m, 1H), 1.99 - 1.84 (m, 1H), 1.14 - 0.97 (m, 1H), 0.84 - 0.74 (m, 1H); MS (ES+): 564.0 (M+l), (ES-): 562.0 (M-l);

Analysis calculated for C₂₅H₂₂BrN₇O4.HC1.2.75H₂O: C, 46.17; H, 4.42; Cl, 5.45; N, 15.08; Found: C, 46.19; H, 4.27; Cl, 5.68; N, 14.87.

Scheme 40

CC^Me

40a

Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-520 methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxylate (40a)

Compound 40a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (39e) (100 mg, 0.241 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-225 yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (80 mg, 0.241 mmol), HATU

137 (110 mg, 0.290 mmol), DIPEA (156 mg, 1.207 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxylate (40a) (53 mg, 78% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.77 (s, 1H, D2O exchangeable), 9.14 (s, 1H), 8.79 - 8.52 (m, 3H, 2H D2O exchangeable), 8.13 (dd, J= 8.7, 1.6 Hz, 1H), 8.00 (d, J= 8.2 Hz, 1H), 7.78 (d, J= 8.7 Hz, 1H), 7.69 (t, J= 8.0

Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.75 (d, J= 17.3 Hz, 1H), 5.39 (d, J= 17.3 Hz, 1H), 4.37 (dd, J= 9.0, 5.9 Hz, 1H), 3.92 (s, 3H), 3.69 (dd, J= 5.7, 2.4 Hz, 1H), 2.56 - 2.42 (m, 1H), 1.98 (dd, J= 13.2, 5.9 Hz, 1H), 1.31 (s, 3H), 1.05-0.97 (m, 1H), 0.97-0.90 (m, 1H); MS (ES+): 578.0 (M+l), (ES-): 576.0 (M-l); Analysis calculated for

C2₆H24BrN₇O₄.HC1.2.25H2O: C, 47.65; H, 4.54; Cl, 5.41; N, 14.96; Found: C, 47.54; H, 4.50;

Cl, 5.29; N, 14.92.

Scheme 41

41f

Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-7-carboxylate (41f)

138

Step-1: Préparation of methyl 2-amino-3-cyano-lH-indole-6-carboxylate (41b)

Compound 41b was prepared according to the procedure reported in step-1 of scheme-11, from methyl 4-iodo-3-(2,2,2-trifluoroacetamido)benzoate (41a) (6.73 g, 18.04 mmol; CAS # 494799-11-0) in DMSO (20 mL) and using malononitrile (1.430 g, 21.65 mmol), L-proline (0.415 g, 3.61 mmol), Cul (0.344 g, 1.804 mmol), K2CO3 (4.99 g, 36.1 mmol) and heating at °C for 16 h under an argon atmosphère. This gave after workup and purification [S1O2 gel (40 g), eluting with EtOAc in hexane from 0-50%] methyl 2-amino-3-cyano-lH-indole-6carboxylate (41b) (1.32 g, 34% yield) as a brown solid; ’H NMR (300 MHz, DMSO-tZô) δ 10.96 (s, 1H), 7.74 (d, J= 1.5 Hz, 1H), 7.63 (dd, J= 8.2, 1.5 Hz, 1H), 7.26 - 7.12 (m, 3H),

3.81 (s, 3H); MS (ES+): 216 (M+l), (ES-): 214 (M-l).

Step-2: Préparation of methyl 4-amino-9H-pyrimido[4,5-b]indole-7-carboxylate (41c)

Compound 41c was prepared according to the procedure reported in step-1 of scheme-6, from methyl 2-amino-3-cyano-lH-indole-6-carboxylate (41b) (1.32 g, 6.13 mmol) using trimethyl orthoformate (13.02 g, 123 mmol), AcOH (1.754 mL, 30.7 mmol) andMLOAc (2.364 g, 15 30.7 mmol). This gave after workup methyl 4-amino-9H-pyrimido[4,5-b]indole-7carboxylate (41c) (0.83 g, 56 % yield) as apale-yellow solid. *HNMR (300 MHz, DMSO-îZô) δ 12.14 (s, 1H), 8.43 (d, J = 8.3 Hz, 1H), 8.30 (s, 1H), 8.03 (d, J = 1.4 Hz, 1H), 7.82 (dd, J = 8.3, 1.5 Hz, 1H), 7.41 (s, 2H), 3.90 (s, 3H); MS (ES+): 243 (M+l), (ES-): 241 (M-l).

Step-3: Préparation of methyl 4-amino-9-(2-(ierZ-butoxy)-2-oxoethyl)-9H-pyrimido[4,520 b]indole-7-carboxylate (41d)

Compound 41d was prepared according to the procedure reported in step-1 of scheme-1, from methyl 4-amino-9H-pyrimido[4,5-b]indole-7-carboxylate (41c) (0.83 g, 3.43 mmol) in DMF (20 mL) using ZerZ-butyl 2-bromoacetate (0.668 g, 3.43 mmol), CS2CO3 (1.340 g, 4.11 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] methyl 4-amino9-(2-(tert-butoxy)-2-oxoethyl)-9H-pyrimido[4,5-b]indoIe-7-carboxylate (41d) (0.67 g, 55% yield) as a pale orange solid; Ή NMR (300 MHz, DMSO-dô) δ 8.48 (d, J= 8.3 Hz, 1H), 8.35 (s, 1H), 8.18 (d, J= 1.5 Hz, 1H), 7.89 (dd, J= 8.2, 1.4 Hz, 1H), 7.54 (s, 2H), 5.23 (s, 2H), 3.91 (s, 3H), 1.41 (s, 9H); MS (ES+): 357 (M+l), (ES-): 355 (M-l).

Step-4: Préparation of 2-(4-amino-7-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (41e)

139

Compound 41e was prepared according to the procedure reported in step-2 of scheme-1, from methyl 4-amino-9-(2-(7erZ-butoxy)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-7-carboxylate (41d) (0.67 g, 1.880 mmol) using TFA (4.29 g, 37.6 mmol) in DCM (20 mL) and stirring at RT for 16 h. This gave after workup 2-(4-amino-7-(methoxycarbonyl)-9H-pyrimido[4,55 b]indol-9-yl)acetic acid (41e) (3.70 g) TFA sait as a yellow solid; ’H NMR (300 MHz, DMSO-àfe) δ 8.71 - 8.47 (m, 4H), 8.36 (d, J= 1.4 Hz, 1H), 7.99 (dd, J= 8.3, 1.4 Hz, 1H), 5.36 (s, 2H), 3.93 (s, 3H); MS (ES+): 301 (M+l), (ES-): 299 (M-l).

Step-5: Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yI)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-710 carboxylate (41f)

Compound 41f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (41e) (50 mg, 0.121 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (38.5 mg, 0.121 mmol), HATU (55.1 mg,

0.145 mmol), DIPEA (78 mg, 0.603 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-220 azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-7-carboxylate (41f) (48 mg, 71% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-t/₆) δ 10.77 (s, 1H, D₂O exchangeable), 8.59 (d, J= 8.5 Hz, 1H), 8.57 (s, 1H), 8.45 (s, 2H, D₂O exchangeable), 8.28 (s, 1H), 8.05 - 7.92 (m, 2H), 7.70 (t, J= 8.0 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.86 (d, J = 17.4 Hz, 1H), 5.49 (d, J= 17.3 Hz, 1H), 4.42 (dd, J= 9.1, 5.6 Hz, 1H), 3.92 (s, 4H), 2.39 25 2.29 (m, 1H), 2.29 - 2.16 (m, 1H), 2.01 - 1.87 (m, 1H), 1.15 - 1.03 (m, 1H), 0.76 - 0.69 (m,

1H); MS (ES+): 564/566 (M+l), (ES-): 562/564 (M-l); Analysis calculated for

C₂₅H₂₂BrN7O4.1.1.HC1.2.75.H₂O: C, 45.91; H, 4.41; Cl, 5.96; N, 14.99; Found: C, 45.90; H, 4.21; Cl, 5.98; N, 14.69.

Scheme 42

140

Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5methyl-2-azabicyclo[3.1.0]hexan-2-yI)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-7-carboxylate (42a)

Compound 42a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-(methoxycarbonyI)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (41e) (100 mg, 0.241 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (80 mg, 0.241 mmol), HATU (110 mg, 0.290 mmol), DIPEA (156 mg, 1.207 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-7-carboxylate (42a) (82 mg, 59% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSOX) δ 10.77 (s, 1H, D2O exchangeable), 8.70 - 8.41 (m, 4H, 2H D2O exchangeable), 8.29 (d, J= 1.4 Hz, 1H), 8.06 - 7.92 (m, 2H), 7.69 (t, J= 7.9 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.83 (d, J= 17.4 Hz, 1H), 5.43 (d, J= 17.3 Hz, 1H), 4.38 (dd, J= 9.0, 5.9 Hz, 1H), 3.92 (s, 3H), 3.73 - 3.70 (m, 1H), 2.54-2.44 (m, 1H), 1.99 (m, 1H), 1.32 (s, 3H), 1.09-1.01 (m, 1H), 0.93 -0.85 (m,

1H); MS (ES+): 578/580 (M+l), (ES-): 576/578 (M-l); Analysis calculated for

C₂₆H24BrN₇O4.HC1.2.5.H2O: C, 47.32; H, 4.58; Cl, 5.37; N, 14.86; Found: C, 47.48; H, 4.28; Cl, 5.23; N, 14.68.

Scheme 43

141

LiOH

43a

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-7-carboxylic acid (43a)

Compound 43a was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-7-carboxylate (41 f) (78 mg, 0.138 mmol) in THF (1 mL) and water (2 mL) using 2M aqueous lithium hydroxide hydrate (0.138 mL, 0.276 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [Cl8 column (100 g), eluting with

ACN in water (containing 0.1% HCl) from 0-100%] 4-amino-9-(2-((lR,3S,5R)-3-((6bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5b]indole-7-carboxylic acid (43a) (34 mg, 45% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-îZô) δ (a mixture of two rotamers) 10.85 (s) and 10.77 (s) (2s, 1H, D₂O exchangeable), 8.93 (s, 2H, D₂O exchangeable), 8.69 (d, J= 4.9 Hz, 1H), 8.63 (d, J= 8.3 Hz,

1H), 8.31 (s, 1H), 7.99 (d, J= 7.9 Hz, 2H), 7.67 (q, J= 7.7 Hz, 1H), 7.29 (dd, J= Ί.Ί, 4.7 Hz,

1H), 5.87 (dd, J= 17.4, 11.5 Hz, 1H), 5.50 (dd, J= 17.3, 7.3 Hz, 1H), 4.42 (dd, J= 9.0, 5.5 Hz, 1H), 3.95-3.92 (m, 1H), 2.39-2.28 (m, 1H), 2.26-2.14 (m, 1H), 2.01 - 1.86 (m, 1H), 1.28 - 1.17 (m) and 1.16 - 1.04 (m) (2m, 1H), 1.02 - 0.83 (m) and 0.79 - 0.67 (m) (2m, 1H); MS (ES+): 550/552 (M+l), (ES-): 548/550 (M-l); Analysis calculated for

C₂₄H₂oBrN₇04.1.1HC1.3H₂0: C, 44.72; H, 4.24; Cl, 6.05; N, 15.21; Found: C, 44.67; H, 4.12;

Cl, 6.07; N, 15.23.

Scheme 44

142

CO2MG

39f ^44a

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxylic acid (44a)

Compound 44a was prepared according to the procedure reported in step-4 of scheme-17, from TFA sait of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxylate (39f) (156mg, 0.276 mmol) in in THF (2 mL) and water (4 mL) using 2M aqueous lithium hydroxide hydrate (0.553 mL, 0.553 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [Cl 8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%) 4-amino-9-(2-((lR,3S,5R)3-((6-bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9Hpyrimido[4,5-b]indole-6-carboxylic acid (44a) (31 mg, 20% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-t/e) (a mixture of two rotamers) δ 12.98 (s, 1H, D2O exchangeable), 10.86 and 10.77 (2s, 1H, D2O exchangeable), 9.11 (s, 1H), 8.60 and 8.58 (2s,

1H), 8.48 (s, 2H, D₂O exchangeable), 8.11 (dt, J= 8.6, 2.1 Hz, 1H), 8.01 and 7.96 (2d, J=

8.3 Hz, 1H), 7.78 - 7.62 (m, 2H), 7.38 - 7.22 (m, 1H), 5.79 (d, J= 17.3 Hz, 1H), 5.44 (d, J= 17.4 Hz, 1H), 4.42 (dd, J= 9.1, 5.5 Hz, 1H), 3.91 (m, 1H), 2.40-2.12 (m, 2H), 1.93 (m, 1H), 1.30 - 0.53 (m, 2H). MS (ES+): 550.1 (M+l); (ES-): 548.0 (M-l); Analysis calculated for C₂4H2oBrN₇04 2.25H₂O.0.95HCl: C, 46.08; H, 4.10; Cl, 5.38; N, 15.67; Found: C, 46.42; H,

4.03; Cl, 4.94; N, 15.29.

Scheme 45

143

Préparation of ethyl 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6-yl)propanoate (45d)

Step-1: Préparation of (E)-ethyl 3-(4-amino-9-(2-(tert-butoxy)-2-oxoethyl)-9H-pyrimido[4,55 b]indol-6-yl)acrylate (45a)

A mixture of tert-butyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (34c) (200 mg, 0.53 mmol), ethyl acrylate (80 mg, 0.795 mmol), Pd(PPh3)2Ch (37.2 mg, 0.053 mmol), and K2CO3 (220 mg, 1.591 mmol) were suspended in DMF (4 mL) in a sealed scintillation vial. The vial was flushed with nitrogen and the yellow mixture then heated at 100 °C for 16 10 h. The resulting black mixture was filtered. The filtrate was diluted with H2O (25 mL) and extracted with EtOAc (25 mL x 3). The combined extract was washed with H2O (25 mL x 4), brine (25 mL), dried, filtered and concentrated in vacuum. The residue obtained was purified by flash column chromatography (silica gel, 24 g, eluting with 0-60% EtOAc in hexane) to provide (E)-ethyl 3-(4-amino-9-(2-(tert-butoxy)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-675 yl)acrylate (45a) (102 mg, 49 % yield) as a pale-yellow solid. *H NMR (300 MHz, DMSOίΖ₆) δ 8.78 (s, 1H), 8.32 (s, 1H), 7.79 (d, J = 15.9 Hz, 1H), 7.73 (d, J = 8.5 Hz, 1H), 7.61 (d, J = 8.5 Hz, 1H), 7.48 (s, 2H), 6.86 (d, J = 15.9 Hz, 1H), 5.15 (s, 2H), 4.22 (q, J = 7.1 Hz, 2H), 1.41 (s, 9H), 1.29 (t, J = 7.1 Hz, 3H); MS (ES+): 397 (M+l), (ES-): 395 (M-l).

Step-2: Préparation of ethyl 3-(4-amino-9-(2-(tert-butoxy)-2-oxoethyl)-9H-pyrimido[4,520 b]indol-6-yl)propanoate (45b)

144

To a solution of (E)-ethyl 3-(4-amino-9-(2-(te/Ÿ-butoxy)-2-oxoethyl)-9H-pyrimido[4,5b]indol-6-yl)acrylate (45a) (100 mg, 0.252 mmol) in THF/ethanol (15 mL, 1:2), was added palladium hydroxide on carbon (7 mg) and heated under a hydrogen atmosphère for 16 h at 100 °C. The reaction mixture was cooled to room température filtered through Celite, washed 5 with ethyl acetate and concentrated in vacuum to dryness. The residue obtained was purified by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM 0-3%] to afford ethyl 3-(4-amino-9-(2-(ter/-butoxy)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6yl)propanoate (45b) (80 mg, 80 % yield) as a white semisolid; *H NMR (300 MHz, DMSOde) δ 8.25 (d, J= 9.1 Hz, 2H), 7.44 (d, J= 8.3 Hz, 1H), 7.36 - 7.14 (m, 3H), 5.08 (s, 2H), 10 4.06 (q, J= 7.1 Hz, 2H), 3.01 (t, J= 7.8 Hz, 2H), 2.75 (t, J= 7.8 Hz, 2H), 1.40 (s, 9H), 1.17 (t, J= 7.2 Hz, 3 H); MS (ES+): 399 (M+l).

Step-3: Préparation of 2-(4-amino-6-(3-ethoxy-3-oxopropyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (45c)

Compound 45c was prepared according to the procedure reported in step-2 of scheme-1, from 15 ethyl 3-(4-amino-9-(2-(ter/-butoxy)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6-yl)propanoate (45b) (78 mg, 0.196 mmol) in DCM (5 mL) using TFA (223 mg, 1.96 mmol) and stirring at RT for 16 h. This gave after workup 2-(4-amino-6-(3-ethoxy-3-oxopropyl)-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (45c) TFA sait as a pale yellow solid; ^!H NMR (300 MHz, DMSO-Jô) δ 8.55 (s, 1H), 8.41 (d, J= 10.5 Hz, 2H), 7.68 (d, J= 8.4 Hz, 1H), 7.43 (d, J 20 = 8.4 Hz, 1H), 5.24 (s, 2H), 4.06 (q, J= 7.1 Hz, 3H), 3.04 (t, J= 7.7 Hz, 2H), 2.76 (t, J= 7.8

Hz, 2H), 1.17 (t, J= 7.1 Hz, 3H); ¹⁹F NMR (282 MHz, DMSO-tZ₆) δ -74.25; MS (ES+): 343 (M+l), (ES-): 341 (M-l).

Step-4: Préparation of ethyl 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-625 yl)propanoate (45d)

Compound 45d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(3-ethoxy-3-oxopropyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (45c) (96 mg, 0.21 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (67.0 mg, 0.210 mmol), 30 HATU (96 mg, 0.252 mmol), DIPEA (136 mg, 1.05 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column

145

chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] ethyl 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indoI-6-yl)propanoate (45d) (44 mg, 35% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 10.75 (s, 1H, 5 D2O exchangeable), 8.58 (s, 1H), 8.53 (s, 2H, D2O exchangeable), 8.39 (s, 1H), 8.00 (d, J=

8.1 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.59 (d, J= 8.4 Hz, 1H), 7.43 (dd, J= 8.3, 1.7 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.73 (d, J= 17.3 Hz, 1H), 5.39 (d, J= 17.3 Hz, 1H), 4.41 (dd, J= 9.0, 5.5 Hz, 1H), 4.05 (q, J= 7.1 Hz, 2H), 3.96 - 3.86 (m, 1H), 3.03 (t, J= 7.7 Hz, 2H), 2.74 (t, 7=7.8 Hz, 2H), 2.39 - 2.26 (m, 1H), 2.26 - 2.13 (m, 1H), 1.98-1.84 (m, 1H), 1.16 (t, 7=

7.1 Hz, 3H), 1.10-1.01 (m, 1H), 0.81-0.72 (m, 1H); MS (ES+): 606/608 (M+l), (ES-):

604/606 (M-l); Analysis calculated for C₂8H₂8BrN7O4.1.25.HC1.1.75.H₂O: C, 49.20; H, 4.83; Cl, 6.48; N, 14.34; Found: C, 49.16; H, 4.63; Cl, 6.68; N, 14.22.

Scheme 46

46d (R= Me, Et mixture) ₄g_e

^46f 46g

Préparation of ethyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylate (46g)

146

Step-1: Préparation of methyl 2-bromo-3-(2,2,2-trifluoroacetamido)benzoate (46b)

To a solution of methyl 3-amino-2-bromobenzoate (46a) (4.5 g, 19.56 mmol; CAS # 10689648-4) and triethylamine (6.82 mL, 48.9 mmol) in DCM (30 mL) was added trifluoracetic anhydride (4.08 mL, 29.3 mmol) dropwise and stirred at RT for 15 h. The reaction mixture was diluted with dichloromethane (75 mL), washed with water (50 mL), dried, filtered and concentrated in vacuum to afford methyl 2-bromo-3-(2,2,2-trifluoroacetamido)benzoate (46b) as a yellow gum (8.17 g) and was used as such for next step; MS (ES+): 325.90 (M+l); (ES-): 323.90 (M-l).

Step-2: Préparation of methyl 2-amino-3-cyano-lH-indole-4-carboxylate (46c)

Compound 46c was prepared according to the procedure reported in step-1 of scheme-11, from methyl 2-bromo-3-(2,2,2-trifluoroacetamido)benzoate (46b) (1594 g, 4.89 mmol) in DMSO (8 mL) using malononitrile (388 mg, 5.87 mmol), L-proline (0.113 g, 0.978 mmol), Cul (93 mg, 0.489 mmol), K2CO3 (1.352 g, 9.78 mmol) in water (8 mL) and heating at 60 °C for 13 h under an argon atmosphère. This gave after workup and purification [S1O2 gel (24 g), eluting with EtOAc in hexane from 0-66%] methyl 2-amino-3-cyano-lH-indole-4carboxylate (46c) (115 mg, 11% yield) as a brown solid; (ES-): 214.00 (M-l).

Step-3: Préparation of methyl/ethyl 4-amino-9H-pyrimido[4,5-b]indoIe-5-carboxylate (46d)

Compound 46d was prepared according to the procedure reported in step-2 of scheme-29, from methyl 2-amino-3-cyano-lH-indole-4-carboxylate (46c) (105 mg, 0.488 mmol) in éthanol (10 mL) using formamidine acetate (513 mg, 4.88 mmol) and heating at 100°C in a microwave .The reaction mixture was filtered, washed with éthanol, and dried under vacuum. The residue was purified by flash column chromatography [S1O2 gel (24 g), eluting with EtOAc in hexane from 0-100% and then using hexanes/10% methanol in ethyl acetate (1:1)] a mixture of methyl/ethyl 4-amino-9H-pyrimido[4,5-b]indole-5-carboxylate (46d) (66 mg) as a brown solid, which was used as such for the next step; MS (ES+): 243.10, 257.10 (M+l).

Step-4: Préparation of methyl/ethyl 4-amino-9-(2-(/erf-butoxy)-2-oxoethyl)-9Hpyrimido[4,5-b]indole-5-carboxylate (46e)

Compound 46e was prepared according to the procedure reported in step-1 of scheme-1, from a mixture of methyl/ethyl 4-amino-9H-pyrimido[4,5-b]indole-5-carboxylate (46d) (65 mg) in

DMF (5 mL) using tert-butyl 2-bromoacetate (0.048 mL, 0.322 mmol), CS2CO3 (219 mg,

147

0.671 mmol) and stirring at RT for 14 h. This gave after workup and purification by flash column chromatography [SiCh gel (24 g), eluting with 10% methanol in ethyl acetate in hexanes from 0-100%] methyl 4-amino-9-(2-(/ert-butoxy)-2-oxoethyl)-9H-pyrimido[4,5b]indole-5-carboxylate (7 mg, 7%); MS (ES+): 357.10 (M+l); ethyl 4-amino-9-(2-(ierZ5 butoxy)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylate (46e) (16 mg, 9% yield) as a white solid; MS (ES+): 371.20 (M+l); (ES-): 369.00 (M-l).

Step-5: Préparation of 2-(4-amino-5-(ethoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (46f)

Compound 46f was prepared according to the procedure reported in step-2 of scheme-1, from 10 ethyl 4-amino-9-(2-(to+-butoxy)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylate (46e) (16 mg, 0.043 mmol) using TFA (0.20 mL, 2.59 mmol) in DCM (5 mL) and stirring at RT for 17 h. This gave after workup 2-(4-amino-5-(ethoxycarbonyl)-9H-pyrimido[4,5-b]indoI-9yl)acetic acid (46f) which was used as such in next step-6 without further purification; MS (ES+): 315.10 (M+l).

Step-6: Préparation of ethyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylate (46g)

Compound 46g was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-5-(ethoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (46f) (13.51mg, 0.043 mmol) in DMF (7 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-220 azabicyclo[3.1.0]hexane-3-carboxamide (4a) (54.8 mg, 0.172 mmol), HATU (65.4 mg, 0.172 mmol) DIPEA (0.045 mL, 0.258 mmol) and stirring at RT for 19 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with hexanes/10% MeOH in EtOAc from 0 - 100%] followed by conversion to HCl sait by dissolving product in acetonitrile (1.5 mL) and 0.1% aq. HCl (6 mL) and lyophilization ethyl 4-amino-9-(225 ((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylate (46g) (6.5 mg, 26% yield) HCl sait as a white solid; 'HNMR(300 MHz, DMSO-7₆) δ 10.76 (s, 1H,), 8.57 (s, 1H), 8.14 -7.93 (m, 4H,), 7.77 - 7.60 (m, 3H), 7.32 (d, J= 7.8 Hz, 1H), 5.82 (d, J= 17.4 Hz, 1H), 5.49 (d, J= 17.3 Hz, 1H), 4.55 - 4.33 (m, 3H), 3.98 - 3.88 (m, 1H), 2.43 - 2.12 (m, 2H), 2.01 - 1.85 (m,

1H), 1.40 (t, J= 7.1 Hz, 3H), 1.15 - 1.02 (m, 1H), 0.84 - 0.73 (m, 1H); MS (ES+): 578.10 &

580.10 (M+l).

148

Scheme 47

(CF₃CO)₂O Et₃N

Cul, K2CO3 cf₃ χ-ΝΗ COOCH₃ ⁰ Br—C ^x>

Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicycIo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-8-carboxylate (47g)

Step-1 : Préparation of methyl 3-bromo-2-(2,2,2-trifluoroacetamido)benzoate (47b)

Compound 47b was prepared according to the procedure reported in step-1 of scheme-46, from methyl 2-amino-3-bromobenzoate (47a) (5 g, 21.73 mmol; CAS # 104670-74-8) in DCM (30 mL) using triethylamine (7.57 mL, 54.3 mmol), trifluoroacetic acid anhydride (4.53 mL, 32.6 mmol) and stirring at RT for 43 h. This gave after workup and purification by 10 flash column chromatography [S1O2 gel (120 g), EtOAc in hexane from 0-14%] methyl 3bromo-2-(2,2,2-trifluoroacetamido)benzoate (47b) as a yellow solid (5.27 g, 74% yield) and was used as such for next step; *H NMR (300 MHz, DMSO-î/ô) δ 11.40 (s, 1H), 8.03 (dd, J= 8.1, 1.4 Hz, 1H), 7.91 (dd, .7=7.8, 1.4 Hz, 1H), 7.48 (t, J = 7.9 Hz, 1H), 3.80 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-ri₆) δ -74.44; MS (ES-): 323.90 & 325.90 (M-l).

Step-2: Préparation of methyl 2-amino-3-cyano-lH-indole-7-carboxylate (47c)

149

Compound 47c was prepared according to the procedure reported in step-1 of scheme-11, from methyl 3-bromo-2-(2,2,2-trifluoroacetamido)benzoate (47b) (2 g, 6.13 mmol) in DMSO (12 mL), using malononitrile (0.486 g, 7.36 mmol), L-proline (0.141 g, 1.227 mmol), Cul (0.117 g, 0.613 mmol), a solution of K2CO3 (1.695 g, 12.27 mmol) in water (12 mL) and 5 heating at 60 °C for 13 h under an argon atmosphère. This gave after workup and purification by flash column chromatography [S1O2 gel (24 g), eluting with EtOAc in hexane from 033%] methyl 2-amino-3-cyano-lH-indole-7-carboxyIate (47c) (305 mg, 23% yield) as a brown solid; Ή NMR (300 MHz, DMSO-ûfc) δ 10.75 (s, 1H), 7.51 (dd, J= 7.8, 1.1 Hz, 1H), 7.41 (d, J= 7.6 Hz, 1H), 7.09 (t, J= 7.7 Hz, 1H), 6.93 (s, 2H), 3.91 (s, 3H); MS (ES+):

216.00.

Step-3: Préparation of methyl 4-amino-9H-pyrimido[4,5-b]indole-8-carboxylate (47d) Compound 47d was prepared according to the procedure reported in step-2 of scheme-29, from methyl 2-amino-3-cyano-lH-indole-7-carboxylate (47c) (300 mg, 1.394 mmol) in éthanol (10 mL) using formamidine acetate (1466 mg, 13.94 mmol) and heating at reflux for 15 45 h. This gave after workup a grey solid residue (268 mg) which was used as such for the next step; Ή NMR (300 MHz, DMSO-î/₆) δ 8.59 (dd, J= 7.8, 1.2 Hz, 1H), 8.34 (s, 1H), 7.97 (dd, .7=7.8, 1.1 Hz, 1H), 7.41 - 7.28 (m, 3H), 3.97 (s, 3H); MS (ES+): 243.10.

Step-4: Préparation of methyl 4-amino-9-(2-(tert-butoxy)-2-oxoethyl)-9H-pyrimido[4,5b]indole-8-carboxylate (47e)

Compound 47e was prepared according to the procedure reported in step-1 of scheme-1, from methyl 4-amino-9H-pyrimido[4,5-b]indole-8-carboxylate (47d) (240 mg, 0.991 mmol) in DMF (6 mL) using ZerZ-butyl 2-bromoacetate (0.176 mL, 1.189 mmol), CS2CO3 (807 mg, 2.477 mmol) and stirring ovemight at RT. This gave after workup and purification by flash column chromatography [S1O2 gel (24 g), eluting with 10% methanol in ethyl acetate in hexanes from 0-100%] methyl 4-amino-9-(2-(tert-butoxy)-2-oxoethyl)-9H-pyrimido[4,5b]indole-8-carboxylate (47e) (353 mg, 25% yield) as an off white solid; *H NMR (300 MHz, DMSO-î/ô) δ 8.61 (dd, J= Ί.9, 1.2 Hz, 1H), 8.36 (s, 1H), 7.84 - 7.79 (m, 1H), 7.49 (s, 2H), 7.37 (t, J= 7.8 Hz, 1H), 5.35 (s, 2H), 3.90 (s, 3H), 1.36 (s, 9H); MS (ES+): 357.15. Step-5: Préparation of 2-(4-amino-8-(methoxycarbonyl)-9H-pyrimido[4,5-b]indoI-9-yl)acetic 30 acid (47f)

Compound 47f was prepared according to the procedure reported in step-2 of scheme-1, from methyl 4-amino-9-(2-(tert-butoxy)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-8-carboxylate (47e) (80 mg, 0.224 mmol) using TFA in DCM (10 mL) and stirring at RT. This gave after

150 workup 2-(4-amino-8-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (47f) which was used as such in next step-6 without purification; MS (ES+): 301.10 (M+l).

Step-6: Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-85 carboxylate (47g)

Compound 47g was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-8-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)açetic acid (47f) (67.3 mg, 0.224 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (71.4 mg, 0.224 mmol), HATU (170 mg, 0.448 10 mmol), DIPEA (0.195 mL, 1.12 mmol) and stirring at RT for 18 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5 %] followed by conversion to HCl sait by dissolving product in acetonitrile (2 mL) and 0.1% aq. HCl (8 mL) and lyophilization methyl 4-amino-9-(2-((lR,3S,5R)-3-((6bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,515 b]indole-8-carboxylate (47g) (38 mg, 30% yield) HCl saltas awhite solid; *H NMR (300 MHz,

DMSO-cfe) δ 10.72 (s, 1H, D₂O exchangeable), 8.65 (d, J= 7.7 Hz, 1H), 8.51 (s, 1H), 8.17 7.91 (m, 3H, 2H D2O exchangeable), 7.82 (d, J= 7.7 Hz, 1H), 7.69 (t, J= 7.9 Hz, 1H), 7.41 (t, J= 7.7 Hz, 1H), 7.30 (d, J= 7.7 Hz, 1H), 5.95 (d, J= 17.5 Hz, 1H), 5.64 (d, J= 17.3 Hz, 1H), 4.39 - 4.27 (m, 1H), 3.92 (s, 3H), 3.87 - 3.72 (m, 1H), 2.37 - 2.09 (m, 2H), 1.97 - 1.78 20 (m, 1H), 1.16 - 0.99 (m, 1H), 0.83 - 0.65 (m, 1H); MS (ES+): 564.10 & 566.10 (M+l); MS (ES-): 562.10 & 564.10 (M-l).

Scheme 48

Préparation of /erZ-butyl (4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo [3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido [4,5 -b] indol-6-yl)carbamate (48d)

Step-1: Préparation of ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a)

Compound 48a was prepared according to the procedure reported in step-2 of scheme-16, from 6-bromo-9H-pyrimido[4,5-b]indol-4-amine (34b) (540 mg, 2.053 mmol) in DMF (10 mL) using ethyl 2-bromoacetate (0.228 mL, 2.053 mmol), CS2CO3 (802 mg, 2.463 mmol), triethylamine (1.716 mL, 12.32 mmol) and stirring at RT for 27 h. This gave after workup 2(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (166 mg) as a lightyellow solid; Ή NMR (300 MHz, DMSO-J₆) δ 8.63 (d, J- 1.8 Hz, 1H), 8.29 (s, 1H), 7.60 (d, J= 8.7 Hz, 1H), 7.54 (dd, J= 8.7, 1.8 Hz, 1H), 7.45 (s, 2H), 5.23 (s, 2H), 4.13 (q, J= 7.1 Hz, 2H), 1.19 (t, J = 7.0 Hz, 3H); MS (ES+): 349.00 & 350.90 (M+l).

Step-2: Préparation of ethyl 2-(4-amino-6-((to7-butoxycarbonyl)amino)-9H-pyrimido[4,5b]indol-9-yl)acetate (48b)

Compound 48b was prepared according to the procedure reported in step-3 of scheme-17, from ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (150 mg, 0.430 mmol) in toluene (8 mL) using XPhos, t-butyl carbamate, Pd2(dba)3, CS2CO3 (140 mg, 0.43 mmol) and heating at 90 °C for 20 h. This gave after work up and purification ethyl 2-(4amino-6-((ter/-butoxycarbonyl)amino)-9H-pyrimido[4,5-b]indol-9-yl)acetate (48b) (10 mg,

6% yield); MS (ES+): 386.20 (M+l).

152

Step-3 : Préparation of 2-(4-amino-6-((ZerZ-butoxycarbonyl)amino)-9H-pyrimido[4,5-b]indol9-yl)acetic acid (48c)

Compound 48c was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-6-((ter/-butoxycarbonyl)amino)-9H-pyrimido[4,5-b]indol-9-yl)acetate (48b) (10 mg, 0.026 mmol) in THF (2 mL) and MeOH (2 mL) using lithium hydroxide hydrate (6.67 mg, 0.156 mmol) and stirring at RT for 18 h. This gave after work up 2-(4amino-6-((ZerZ-butoxycarbonyI)amino)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (48c) which was used as such in next step-4; MS (ES+): 358.20 (M+l); (ES-): 356.10 (M-l).

Step-4: Préparation of ZerLbutyl (4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-210 yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6yl)carbamate (48d)

Compound 48d was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-((Zert-butoxycarbonyl)amino)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (48c) (from above step-3, 0.026 mmol) in DMF (6 mL) using HCl sait of (lR,3S,5R)-N-(615 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (71 mg, 0.224 mmol),

HATU (40 mg, 0.104 mmol), DIPEA (0.027 mL, 0.156 mmol) and stirring at RT for 14 h. This gave after workup and purification by flash column chromatography [silica gel (4 g), eluting with MeOH in DCM from 0-5 %] followed by conversion to HCl sait in acetonitrile (1 mL) using 0.1% aq. HCl (5 mL) and lyophilization ZerZ-butyl (4-amino-9-(2-((lR,3S,5R)20 3-((6-bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9Hpyrimido[4,5-b]indol-6-yl)carbamate (48d) (6 mg, 37% yield) HCI sait as a white solid; 'H NMR (300 MHz, DMSO-î/₆) δ 10.76 (s, 1H, D₂O exchangeable), 9.26 (s, 1H, D₂O exchangeable), 8.54 (s, 1H), 8.38 (s, 1H), 8.29 (s, 2H, D₂O exchangeable), 8.01 (d, J= 8.1 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.56 (d, J= 8.8 Hz, 1H), 7.40 (d, J= 8.1 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.70 (d, J= 17.3 Hz, 1H), 5.37 (d, J= 17.2 Hz, 1H), 4.41 (dd, J= 9.1, 5.5 Hz,

1H), 3.96-3.85 (m, 1H), 2.44-2.07 (m, 2H), 1.98-1.81 (m, 1H), 1.49 (s, 9H), 1.13-0.98 (m, 1H), 0.85-0.62 (m, 1H); MS (ES+): 621.20 & 623.20 (M+l); MS (ES-): 619.10 & 621.10 (M-l).

, Scheme 49

153

48a 49a 49b ₄g_c

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-cyano-9H-pyrimido[4,5-b]indol-9-yI)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (49c)

Step-1: Préparation of ethyl 2-(4-amino-6-cyano-9H-pyrimido[4,5-b]indol-9-yl)acetate (49a)

To a mixture of ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (300 mg, 0.859 mmol) and dicyanozinc (303 mg, 2.58 mmol) in DMF (8 mL) was added Pd(PPh3)₄ (149 mg, 0.129 mmol), degassed, fïlled with nitrogen, and heated at 100 °C for 13 h. The reaction mixture was diluted with ethyl acetate (120 mL), washed with water (2 x 60 mL), brine (60 mL), dried, filtered, concentrated in vacuum and triturated with ethyl acetate (20 mL). The solid obtained was collected by filtration and dried in vacuum to afford ethyl 2(4-amino-6-cyano-9H-pyrimido[4,5-b]indol-9-yl)acetate (49a) (160 mg, 63% yield) as a white solid; 1H NMR (300 MHz, DMSO-tZ₆) δ 8.94 - 8.93 (m, 1H), 8.36 (s, 1H), 7.86 - 7.80 (m, 2H), 7.59 (s, 2H), 5.31 (s, 2H), 4.14 (q, J = 7.1 Hz, 2H), 1.20 (t, J= 7.2 Hz, 3H); MS (ES+): 296.10 (M+l); (ES-): 294.10 (M-l)

Step-2: Préparation of 2-(4-amino-6-cyano-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (49b)

Compound 49b was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-6-cyano-9H-pyrimido[4,5-b]indol-9-yl)acetate (49a) (80 mg, 0.271 mmol) in THF (3 mL) and MeOH (3 mL) using a solution of lithium hydroxide hydrate (69.6 mg, 1.625 mmol) in water (3 mL) and stirring at RT for 15 h. This gave after workup 2-(420 amino-6-cyano-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (49b) (72 mg, 100% yield) which was used as such in next step-3 without further purification; MS (ES+) : 2£8. 10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-cyano-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (49c)

Compound 49c was prepared according to the procedure reported in step-3 of scheme-1, from 25 2-(4-amino-6-cyano-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (49b) (0.072 g, 0.271 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-221436

154 azabicyclo[3.1.0]hexane-3-carboxamide (4a) (0.104 g, 0.325 mmol), HATU (0.206 g, 0.542 mmol), DIPEA (175 mg, 1.355 mmol) and stirring at RT for 22 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), 10% MeOH in EtOAc in hexane from 0 - 100 %] followed by conversion to HCl sait in acetonitrile (5 mL) using 0.1% aq. HCl (30 mL) and lyophilization (lR,3S,5R)-2-(2-(4-amino-6-cyano-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (49c) (54 mg, 38% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-7₆) δ 10.76 (s, 1H, D₂O exchangeable), 9.09 (d, 7= 1.5 Hz, 1H), 8.74 - 8.54 (m, 3H, 2H D₂O exchangeable), 8.00 (d, J= 8.2 Hz, 1 H), 7.94 (dd, J= 8.6, 1.4 Hz, 1H), 7.87 (d, J= 8.6 Hz, 1H), 7.70 (t, J =8.0 Hz, 1H), 7.31 (d, 7= 7.7 Hz, 1H), 5.81 (d, 7= 17.4 Hz, 1 H), 5.45 (d,7 = 17.3 Hz, 1H), 4.42 (dd, 7= 9.0, 5.5 Hz, 1H), 3.95 - 3.84 (m, 1H), 2.42 - 2.08 (m, 2H), 1.98 1.83 (m, 1H), 1.16-0.98 (m, 1H), 0.90-0.67 (m, 1H); MS (ES+): 531.10 & 533.10 (M+l); MS (ES-): 529.00 & 531.10 (M-l); Analysis calculated for C₂₄Hi₉BrN₈0₂.0.85HC1.2.0H₂0: C, 48.17; H, 4.02; Cl, 5.04; N, 18.73; Found: C, 48.54; H, 4.17; Cl, 5.36; N, 18.40.

Scheme 50

Cul, K2CO3

50f

Préparation of (lR,3S,5R)-2-(2-(4-amino-5-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (50f)

Step-1: Préparation of 2-amino-4-fluoro-lH-indole-3-carbonitrile (50b)

Compound 50b was prepared according to the procedure reported in step-1 of scheme-11, fromN-(2-bromo-3-fluorophenyl)-2,2,2-trifluoroacetamide (50a) (5 g, 17.48 mmol; CAS # 118313-91-0) in DMSO (20 mL) using malononitrile (1.386 g, 20.98 mmol), L-proline

155 (0.403 g, 3.50 mmol), Cul (0.333 g, 1.748 mmol), K2CO3 (4.83 g, 35.0 mmol) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification by flash column chromatography [S1O2 gel (40 g), eluting with EtOAc in hexane from 0-50%] 2amino-4-fluoro-lH-indole-3-carbonitrile (50b) (0.3 g, 10% yield) as an brown solid; ’H

NMR (300 MHz, DMSO-îZ₆) δ 10.97 (s, 1H), 6.98 (d, J= 7.8 Hz, 1H), 6.91 - 6.83 (m, 3H), 6.75 (ddd, J= 10.8, 8.1, 1.0 Hz, 1H); MS (ES+): 176.0 (M+l), (ES-): 174.0 (M-l).

Step-2: Préparation of 5-fluoro-9H-pyrimido[4,5-b]indol-4-amine (50c)

Compound 50c was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-4-fluoro-lH-indole-3-carbonitrile (50b) (0.3 g, 1.713 mmol) using trimethyl orthoformate (3.64 g, 34.3 mmol), AcOH (0.490 mL, 8.56 mmol) and NH4OAC (0.660 g, 8.56 mmol). This gave after workup and purification by reverse phase column chromatography [Cl8 column (30 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] 5-fhioro-9H-pyrimido[4,5-b]indoI-4-amine (50c) (83 mg, 24% yield) as a brown yellow solid; ΉNMR (300 MHz, DMSO-d₆) δ 13.18 (s, 1H), 8.60 (s, 1H), 7.61 - 7.38 (m,

2H), 7.23 (dd, J= 11.3, 7.8 Hz, 1H); MS (ES+): 203.04 (M+l).

Step-3: Préparation of terZ-butyl 2-(4-amino-5-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (50d)

Compound 50d was prepared according to the procedure reported in step-2 of scheme-16, from HCl sait of 5-fluoro-9H-pyrimido[4,5-b]indol-4-amine (50c) (80 mg, 0.335 mmol) in

DMF (2.5 mL) using ZerZ-butyl 2-bromoacetate (65.4 mg, 0.335 mmol) CS2CO3 (218 mg, 0.670 mmol) and stirring at RT for 1.5 h. The solid separated was collected by filtration, dried to give terributyl 2-(4-amino-5-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (50d) (83 mg, 78 % yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-î/ô) δ 8.36 (s, 1H), 7.54 - 7.40 (m, 2H), 7.14 (ddd, J= 11.5, 7.2, 1.7 Hz, 1H), 5.16 (s, 2H), 1.41 (s, 9H); MS (ES+):

317.10 (M+l).

Step-4: Préparation of 2-(4-amino-5-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (50e)

Compound 50e was prepared according to the procedure reported in step-2 of scheme-1, from fërributyl 2-(4-amino-5-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (50d) (80 mg, 0.253 mmol) using 20% TFA in DCM (1452 μΐ, 3.79 mmol) and stirring at RT for 16 h. This gave after workup 2-(4-amino-5-fluoro-9H-pyrimido[4,5-b]indol-9-yI)acetic acid (50e) (85 mg, 90% yield) TFA sait as a yellow solid; 'H NMR (300 MHz, DMSO-tfc) δ 8.46 (s, 1H), 7.58

156 (d, J= 8.2 Hz, 1H), 7.49 (td, J= 8.1, 5.5 Hz, 1H), 7.20 (dd, J= 11.3, 8.0 Hz, 1H), 5.22 (s, 2H); MS (ES+): 261.00 (M+l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-5-fluoro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (50f)

Compound 50f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-5-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (50e) (50 mg, 0.134 mmol) in DMF (1.5 mL) using TFA sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (52.9 mg, 0.134 mmol), HATU (76 mg, 0.200 mmol), DIPEA (0.116 mL, 0.668 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-5-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (50f) (53 mg, 76% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-ùfe) δ 10.77 (s, 1H, D₂O exchangeable), 8.68 (s, 1H), 8.01 (d, 8.1 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.62 - 7.49 (m, 2H), 7.35 7.20 (m, 2H), 5.79 (d, J= 17.4 Hz, 1H), 5.44 (d, J= 17.3 Hz, 1H), 4.43 (dd, J= 9.1, 5.5 Hz, 1H), 3.91 (ddd, J= 7.4, 5.4, 2.3 Hz, 1H), 2.40 - 2.14 (m, 2H), 1.92 (dq, J= 13.3, 6.5, 6.0 Hz, 1H), 1.08 (dt, J= 8.7, 5.4 Hz, 1H), 0.79 (td, J= 5.2, 2.4 Hz, 1H); ¹⁹F NMR (282 MHz,

DMSO-Jô) δ -113.23; MS (ES+): 524.0 (M+l); 546.0 (M+Na); (ES-): 522.0 (M-l); Analysis calculated for C23H19BrFN7O2.L75H2O.lHCl: C, 46.64; H, 4.00; Cl, 5.99; N, 16.55; Found: C, 46.50; H, 3.71; Cl, 5.73; N, 16.29.

Scheme 51

157

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo [3.1.0]hexane-3 -carboxamide (51g)

Step-1: Préparation of 2,2,2-trifluoro-N-(2-fluoro-6-iodophenyl)acetamide (51b)

Compound 51b was prepared according to the procedure reported in step-1 of scheme-46, from 2-fluoro-6-iodoaniline (51a) (5 g, 21.10 mmol; CAS # 886762-73-8) in DCM (50 mL) using triethylamine (5 mL, 35.9 mmol), trifluoroacetic acid anhydride (4.40 mL, 6.65 mmol) and stirring at RT for 16 h. This gave aller workup 2,2,2-trifluoro-N-(2-fluoro-6iodophenyl)acetamide (51b) (6.5 g, 93% yield) as a yellow solid, which was used as such for 10 the next step; Ή NMR (300 MHz, DMSO-tZ₆) δ 11.42 (s, 1H), 7.81 (dt, J= 8.0, 1.2 Hz, 1H), 7.43 (ddd, J= 9.6, 8.4, 1.3 Hz, 1H), 7.26 (td, J= 8.2, 5.6 Hz, 1H); MS (ES-): 331.80 (M-l).

Step-2: Préparation of 2-amino-7-fluoro-lH-indole-3-carbonitrile (51c)

Compound 51c was prepared according to the procedure reported in step-1 of scheme-11, from 2,2,2-trifluoro-N-(2-fluoro-6-iodophenyl)acetamide (51b) (6.4 g, 19.22 mmol) in

DMSO (20 mL) using malononitrile (1.452 mL, 23.06 mmol), L-proline (0.443 g, 3.84 mmol), Cul (0.366 g, 1.922 mmol), K2CO3 (5.31 g, 38.4 mmol) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [S1O2 gel (40 g), eluting with EtOAc in hexane from 0-50%] 2-amino-7-fluoro-lH-indole-3-carbonitrile (51c) (1.535

158 g, 46% yield) as a yellow solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 11.33 (s, 1H), 7.03 - 6.88 (m, 2H), 6.88 - 6.66 (m, 3H); MS (ES+): 176.0 (M+l), (ES-): 174.0 (M-l).

Step-3: Préparation of 8-fluoro-9H-pyrimido[4,5-b]indoI-4-amine (51d)

Compound 51d was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-7-fluoro-lH-indole-3-carbonitrile (51c) (1.5 g, 8.56 mmol) using trimethyl orthoformate (18.74 mL, 171 mmol), AcOH (2.449 mL, 42.8 mmol) and NH4OAC (3.30 g, 42.8 mmol). This gave after workup 8-fluoro-9H-pyrimido[4,5-b]indol-4-amine (51d) (1.54 g, 69% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-J₆) δ 12.33 (s, 1H), 11.98 (s, 1H), 8.29 (s, 1H), 8.18 - 8.03 (m, 1H), 7.27 (s, 2H), 7.26 - 7.16 (m, 2H), 1.92 (s, 3H); MS (ES+): 203.10 (M+l).

Step-4: Préparation of fôrZ-butyl 2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (51e)

Compoûnd 51e was prepared according to the procedure reported in step-2 of scheme-16, from AcOH sait of 8-fluoro-9H-pyrimido[4,5-b]indol-4-amine (51d) (1.5 g, 5.72 mmol) in

DMF (35 mL) using fôrt-butyl 2-bromoacetate (1.171g, 6.01 mmol), CS2CO3 (4.10 g, 12.58 mmol) stirring at RT for 15 h, followed by the addition of K2CO3 (0.791 g, 5.72 mmol), tertbutyl 2-bromoacetate (0.845 mL, 5.72 mmol) and stirring for additional 3 h. This gave after workup fërf-butyl 2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (51e) (1.15 g, 64% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-t/₆) δ 8.34 (s, 1H), 8.22 - 8.16 (m, 1H), 7.45 (s, 2H), 7.28 - 7.21 (m, 2H), 5.17 (d, J= 2.0 Hz, 2H), 1.42 (s, 9H); MS (ES+):

317.14 (M+l).

Step-5: Préparation of 2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (51f)

Compound 51f was prepared according to the procedure reported in step-2 of scheme-1, from terZ-butyl 2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (51e) (0.9 g, 2.85 mmol) using 20% TFA in DCM (16.33 mL, 42.7 mmol)) and stirring at RT for 16 h. This gave after workup 2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (51f) (0.92 g, 86% yield) TFA sait as a white solid; Ή NMR (300 MHz, DMSO-ri₆) δ 8.49 (s, 1H), 8.32 - 8.23 (m, 1H), 8.07 (s, 2H), 7.40 - 7.27 (m, 2H), 5.25 (d, J= 1.9 Hz, 2H); MS (ES+): 261.10 (M+l). .

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (51g)

I

159

Compound 51g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (511) (75 mg, 0.200 mmol) in DMF (2 mL) using TFA sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (79 mg, 0.200 mmol), HATU (114 mg, 0.301 mmol), DIPEA (0.175 mL, 1.002 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(610 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (51g) (81 mg, 77% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-îZô) δ 10.78 (s, 1H, D2O exchangeable), 8.60 (s, 1H), 8.45 (s, 2H, D₂O exchangeable), 8.32 (dd, J= 5.8, 3.0 Hz, 1H), 8.02 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.41 - 7.26 (m, 3H), 5.81 (d, J= 17.5 Hz, 1H), 5.57 - 5.38 (m, 1H), 4.44 (dd, J= 9.0, 5.6 Hz, 1H), 3.98 - 3.80 (m, 1H), 2.40 - 2.13 (m, 2H), 2.01-1.83 (m, 1H), 1.10 (dt, J= 9.3, 5.4 Hz, 1H), 0.64 (q, J= 3.8, 2.9 Hz, 1H). ^I9F NMR (282 MHz,

DMSO-î/ô) δ -134.64. MS (ES+): 524.1 (M+l); (ES-): 522.1 (M-l); Analysis calculated for C23Hi9BrFN7O₂.2H₂O.0.9HCl: C, 46.57; H, 4.06; Cl, 5.38; N, 16.53; Found: C, 46.54; H, 3.90; Cl, 5.43; N, 16.32.

Scheme 52

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (52f)

Step-1: Préparation of 2-amino-6-bromo-lH-indole-3-carbonitrile (52b)

160

Compound 52b was prepared according to the procedure reported in step-1 of scheme-11, from N-(5-bromo-2-iodophenyl)-2,2,2-trifluoroacetamide (52a) (6.4 g, 16.25 mmol; CAS # 1870674-39-7) in DMSO (20 mL) using malononitrile (1.228 mL, 19.50 mmol), L-proline (0.374 g, 3.25 mmol), Cul (0.309 g, 1.625 mmol), K2CO3 (4.49 g, 32.5 mmol) and heating at 5 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [SiCh gel (40 g), eluting with EtOAc in hexane from 0-50%] 2-amino-6-bromo-lH-indole-3carbonitrile (52b) (2.06 g, 54% yield) as a yellow solid; ’H NMR (300 MHz, DMSO-cfe) δ 10.80 (s, 1H), 7.29 (d, J= 1.6 Hz, 1H), 7.17 - 7.00 (m, 2H), 6.95 (s, 2H); MS (ES+): 236.0 (M+l), (ES-): 233.9 (M-l).

Step-2: Préparation of 7-bromo-9H-pyrimido[4,5-b]indol-4-amine (52c)

Compound 52c was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-6-bromo-lH-indole-3-carbonitrile (52b) (2 g, 8.47 mmol) using trimethyl orthoformate (17.98 g, 169 mmol), AcOH (2.54 g, 42.4 mmol) and NH4OAC (3.27 g, 42.4 mmol). This gave after workup 7-bromo-9H-pyrimido[4,5-b]indol-4-amine (52c) (2.29 g,

84% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-ûfe) δ 11.97 (s, 2H), 8.27 (t, J = 4.2 Hz, 2H), 7.58 (d, J= 1.8 Hz, 1H), 7.36 (dd, J= 8.4, 1.8 Hz, 1H), 7.26 (s, 2H), 1.92 (s, 3H).

Step-3: Préparation of tert-butyl 2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indoI-9-yl)acetate (52d)

Compound 52d was prepared according to the procedure reported in step-2 of scheme-16, from AcOH sait of 7-bromo-9H-pyrimido[4,5-b]indol-4-amine (52c) (2.2 g, 6.81 mmol) in DMF (75 mL) using tert-butyl 2-bromoacetate (1.056 g, 7.15 mmol), CS2CO3 (4.88 g, 14.98 mmol) stirring at RT for 15 h, followed by the addition of K2CO3 (0.941 g, 6.81 mmol), tertbutyl 2-bromoacetate (1.006 mL, 6.81 mmol) and stirring for 3 h at RT. This gave after workup tert-butyl 2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (52d) (1.25 g, 49% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-J₆) δ 8.31 (t, J= 4.2 Hz, 2H), 7.91 (d, J= 1.7 Hz, 1H), 7.46-7.36 (m, 3H),5.14(s, 2H), 1.41 (s, 9H).

Step-4: Préparation of 2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (52e)

Compound 52e was prepared according to the procedure reported in step-2 of scheme-1, from 30 tert-butyl 2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (52d) (0.3 g, 0.795 mmol) using 20% TFA in DCM (4.56 mL, 11.93 mmol) and stirring at RT for 16 h. This gave after workup 2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (52e) (0.31

161 g, 90% yield) TFA sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 8.49 (s, 1H), 8.41 (d, J= 8.4 Hz, 1H), 8.08 (d, J= 1.7 Hz, 1H), 7.54 (dd, J= 8.4, 1.7 Hz, 1H), 5.23 (s, 2H); MS (ES+): 321.00 (M+l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-95 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (52f)

Compound 52f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (52e) (75 mg, 0.172 mmol) in DMF (2 mL) using TFA sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (68.3 mg, 0.172 mmol), HATU (98 mg, 0.259 mmol), DIPEA (0.150 mL, 0.862 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(675 bromopyridin-2-yl)-2-azabicyclo[3.1,0]hexane-3-carboxamide (52f) (79 mg, 78 % yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-îZô) δ 10.79 (s, 1H, D₂O exchangeable), 8.70 (s, 2H, D₂O exchangeable), 8.64 (s, 1H), 8.49 (d, J= 8.5 Hz, 1H), 8.08 - 7.97 (m, 2H), 7.71 (t, J= 8.0 Hz, 1H), 7.60 (dd, J= 8.5, 1.7 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.79 (d, J = 17.4 Hz, 1H), 5.42 (d, J= 17.3 Hz, 1H), 4.43 (dd, J= 9.0, 5.5 Hz, 1H), 3.90 (ddd, J= 7.6,

5.4, 2.4 Hz, 1H), 2.39-2.10 (m, 2H), 2.01 - 1.83 (m, 1H), 1.08 (dt, J = 8.8, 5.5 Hz, 1H),

0.79 (td, J= 5.2, 2.4 Hz, 1H). MS (ES+): 586.0 (M+l); (ES-): 584.0 (M-l); Analysis calculated for C₂₃Hi₉Br₂N₇O₂.2H₂O.lHCl: C, 42.00; H, 3.68; Cl, 5.39; N, 14.91; Found: C, 42.08; H, 3.55; Cl, 5.18; N, 14.77.

Scheme 53

162

Préparation of (S)-5-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyI)N-(6-bromopyridin-2-yl)-5-azaspiro[2.4]heptane-6-carboxamide (53d).

Step-1: Préparation of (S)-teri-butyl 6-((6-bromopyridin-2-yl)carbamoyl)-55 azaspiro[2.4]heptane-5-carboxylate (53b)

To a stirred solution of (S)-5-(terf-butoxycarbonyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (53a) (0.5 g, 2.072 mmol) in DCM (15 mL) was added 1-methyl-lH-imidazole (0.413 mL, 5.18 mmol) at 0-5 °C under a nitrogen atmosphère. The reaction mixture was stirred for 10 min at 0-5 °C, added methanesulfonyl chloride (0.192 mL, 2.487 mmol) followed by stirring at 0-5 °C for 1 h. To this mixture was added 6-bromopyridin-2-amine (0.359 g, 2.072 mmol) and stirred for 18 h at RT. Water (30 mL) was added to the reaction mixture, layers were separated, and aqueous layer was extracted with DCM (3x30 mL). The combined organics were washed with IN HCl (30 mL), Sat. aqueous NaHCCh (30 mL), brine (30 mL), dried, filtered and concentrated in vacuum to afford (S)-ierZ-butyl 6-((6-bromopyridin-215 yl)carbamoyl)-5-azaspiro[2.4]heptane-5-carboxylate (53b) (0.8 g, 97 % yield); 'H NMR (300 MHz, DMSO-cfo) δ 10.90 (d, J= 18.6 Hz, 1H), 8.11 (dd, J= 13.0, 8.1 Hz, 1H), 7.75 (q, J= 7.8 Hz, 1H), 7.35 (d, J= 7.7 Hz, 1H), 4.57 - 4.39 (m, 1H), 3.32 - 3.16 (m, 2H), 2.15 (ddd, J = 27.3, 12.8, 8.3 Hz, 1H), 1.83 (ddd, J= 35.1, 12.6, 5.2 Hz, 1H), 1.33 (d, J= 36.8 Hz, 9H), 0.68 - 0.41 (m, 4H); MS (ES+): 396.00 (M+l).

Step-2: Préparation of (S)-N-(6-bromopyridin-2-yl)-5-azaspiro[2.4]heptane-6-carboxamide (53c)

163

Compound 53c was prepared according to the procedure reported in step-2 of scheme-1, from (S)-terZ-butyl 6-((6-bromopyridin-2-yl)carbamoyl)-5-azaspiro[2.4]heptane-5-carboxylate (53b) (0.8 g, 2.019 mmol) using TFA (0.778 mL, 10.09 mmol) in DCM (7 mL) and stirring ovemight at RT. This gave after workup (S)-N-(6-bromopyridin-2-yl)-55 azaspiro[2.4]heptane-6-carboxamide (53c) (0.55 g, 92% yield) TFA sait; *H NMR (300 MHz, DMSO-ί/ό) δ 11.43 (s, 1H), 9.71 (s, 1H), 8.90 (s, 1H), 8.08 (d, J= 8.2 Hz, 1H), 7.83 (t, J= 7.9 Hz, 1H), 7.45 (d, J= 7.7 Hz, 1H), 4.63 - 4.47 (m, 1H), 3.20 (d, J= 5.2 Hz, 2H), 2.33 (dd, J= 13.1, 8.6 Hz, 1H), 2.03 (dd, J= 13.1, 7.0 Hz, 1H), 0.76 - 0.46 (m, 4H); MS (ES+): 295.93 (M+l). .

Step-3: Préparation of (S)-5-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-azaspiro[2.4]heptane-6-carboxamide (53d) Compound 53d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (S)-N-(6-bromopyridin-2-yl)-5-azaspiro[2.4]heptane-6-carboxamide (53c) (72.5 mg, 0.177 mmol) in DMF (2 mL) using TFA sait of 2-(4-amino-6-(trifluorpmethyl)15 9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (75 mg, 0.177 mmol), HATU (101 mg, 0.265 mmol), DIPEA (0.154 mL, 0.884 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (S)-520 (2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yI)acetyl)-N-(6-bromopyridin-2yl)-5-azaspiro[2.4]heptane-6-carboxamide (53d)) (89 mg, 86% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-dô) (a mixture of two rotamers) δ 11.25 and 10.82 (2s, 1H, D2O exchangeable), 8.96 (s, 1H), 8.59 and 8.57 (2s, 1H), 8.52 (s, 2H, D2O exchangeable), 8.19 and 8:02 (2d, J= 8.2 Hz, 1H), 7.94 - 7.78 (m, 2H), 7.70 (t, J= 8.0 Hz, 1H), 7.41 and

7.32 (2d, J= 7.7 Hz, 1H), 5.45 (d, J= 2.4 Hz, 2H), 4.65 (dd, J= 8.5, 5.0 Hz, 1H), 3.90 - 3.74 (m, 2H), 2.25 (dd, J= 12.7, 8.4 Hz, 1H), 1.89 (dd, J= 12.6, 4.9 Hz, 1H), 0.81 - 0.49 (m, 4H). ¹⁹F NMR (282 MHz, DMSO-cfc) δ -58.70. MS (ES+): 588.1 (M+l); (ES-): 586.1 (M-l); Analysis calculated for C25H2iBrF₃N₇O2 1.5H₂O.1.1HC1: C, 45.81; H, 3.86; Cl, 5.95; N, 14.96; Found: C, 45.85; H, 3.73; Cl, 5.91; N, 14.70.

Scheme 54

164

54d

54f

4a

HATU, DIPEA

54e

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicycIo[3.1.0]hexane-3-carboxamide (54g)

Step-1: Préparation of N-(2-bromo-6-methoxyphenyl)-2,2,2-trifluoroacetamide (54b)

Compound 54b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-6-methoxyaniline (54a) (5 g, 24.75 mmol; CAS # 5473-01-8) in DCM (50 mL) using triethylamine (5.86 mL, 42.1 mmol), trifluoroacetic acid anhydride (5.16 mL, 37.1 mmol) and stirring at RT for 16 h. This gave after workup N-(2-bromo-6-methoxyphenyl)2,2,2-trifluoroacetamide (54b) (7.35 g, 100% yield) as a pale yellow solid and was used as such for next step; ’H NMR (300 MHz, DMSO-cfe) δ 11.03 (s, 1H), 7.40 - 7.26 (m, 2H), 7.18 (dd, J= 7.1, 2.5 Hz, 1H), 3.82 (s, 3H); MS (ES-): 295.9 (M-l).

Step-2: Préparation of 2-amino-7-methoxy-lH-indole-3-carbonitrile (54c)

Compound 54c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-6-methoxyphenyl)-2,2,2-trifluoroacetamide (54b) (7.2 g, 24.16 mmol) in

DMSO (20 mL) and using malononitrile (1.826 mL, 29.0 mmol), L-proline (0.556 g, 4.83 mmol), Cul (0.460 g, 2.416 mmol), K2CO3 (6.68 g, 48.3 mmol) and stirring at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [S1O2 gel (40 g), eluting

165 with EtOAc in hexane from 0-50%] 2-amino-7-methoxy-lH-indole-3-carbonitrile (54c) (2.308 g, 51% yield) as a yellow solid; Ή NMR (300 MHz, DMSO-7₆) δ 10.81 (s, 1H), 6.92 (t, J= 7.9 Hz, 1H), 6.77 (d, J= 7.7 Hz, 1H), 6.59 (d, 7= 7.9 Hz, 1H), 6.37 (s, 2H), 3.86 (s, 3H); MS (ES+): 188.1 (M+l); (ES-): 186.0 (M-l).

Step-3: Préparation of 8-methoxy-9H-pyrimido[4,5-b]indoI-4-amine (54d)

Compound 54d was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-7-methoxy-lH-indole-3-carbonitrile (54c) (2.3 g, 12.29 mmol) using trimethyl orthoformate (26.1 g, 246 mmol), AcOH (3.51 mL, 61.4 mmol) andNHiOAc (4.74 g, 61.4 mmol). This gave after workup 8-methoxy-9H-pyrimido[4,5-b]indol-4-amine (54d) (2.7 g,

80% yield) AcOH sait as a pale yellow solid; *H NMR (300 MHz, DMSO-Ts) δ 11.99 (s, 1H),

11.91 (s, 1H), 8.23 (s, 1H), 7.88 (d, 7= 7.8 Hz, 1H), 7.15 (t, 7= 7.9 Hz, 1H), 7.08 (s, 2H), 6.98 (d, J= 7.9 Hz, 1H), 3.95 (s, 3H), 1.91 (s, 3H).

Step-4: Préparation of tert-butyl 2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetate (54e)

Compound 54e was prepared according to the procedure reported in step-2 of scheme-16, from AcOH sait of 8-methoxy-9H-pyrimido[4,5-b]indol-4-amine (54d) (2.6 g, 9.48 mmol) in DMF (75 mL) using terributyl 2-bromoacetate (1.471 mL, 9.95 mmol), CS2CO3 (6.80 g, 20.86 mmol), stirring at RT for 15 h, followed by the addition of tert-butyl 2-bromoacetate (1.401 mL, 9.48 mmol), K2CO3 (1.31 g, 9.48 mmol) and stirring for 3 h. The solid separated was filtered and dried to give /erAbutyl 2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetate (54e) (1.87 g, 60% yield) as a pale yellow solid; ’H NMR (300 MHz, DMSO-Ts) δ 8.28 (s, 1H), 7.94 (d, 7= 7.8 Hz, 1H), 7.26 (s, 2H), 7.20 (t, 7= 7.9 Hz, 1H), 7.01 (d, 7= 8.0 Hz, 1H), 5.19 (s, 2H), 3.89 (s, 3H), 1.42 (s, 9H).

Step-5: Préparation of 2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (54f)

Compound 54f was prepared according to the procedure reported in step-2 of scheme-1, from terributyl 2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (54e) (0.9 g, 2.74 mmol) using 20% TFA in DCM (15.73 mL, 41.1 mmol)) and stirring at RT for 16 h. This gave after workup afford 2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (54i) (0.46 g, 43% yield) TFA sait as a white solid; *H NMR (300 MHz, DMSO-Ts) δ 8.51 (s,

1H), 8.20 (s, 2H), 8.05 (d, 7= 7.9 Hz, 1H), 7.32 (t, 7= 8.0 Hz, 1H), 7.13 (d, 7= 8.0 Hz, 1H),

5.32 (s, 2H), 3.92 (s, 3H).

166

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo [3.1.0]hexane-3-carboxamide (54g)

Compound 54g was prepared according to the procedure reported in step-3 of scheme-1, from TFA saltof 2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (54f) (75 mg,

0.194 mmol) in DMF (2 mL) using TFA sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (77 mg, 0.194 mmol), HATU (111 mg, 0.291 mmol), DIPEA (0.169 mL, 0.971 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography

[C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (54g) (85 mg, 82% yield) as a white solid; Ή NMR (300 MHz, DMSO-cfe) δ 10.74 (s, 1H, D₂O exchangeable), 8.60 (s, 1H), 8.57 (s, 2H, D₂O exchangeable), 8.08 (d, J= 7.9 Hz, 1H), 8.02 (d, J= 8.2 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.40 - 7.24 (m, 2H), 7.15 (d, J= 8.0 Hz, 1H), 5.81 (d, J= 17.0 Hz, 1H), 5.59 (d, J= 16.9 Hz, 1H), 4.43 (dd, J= 8.9, 5.6 Hz, 1H), 3.94 (s, 3H), 3.85 (td, J= 6.3, 5.3, 2.3 Hz, 1H), 2.42-2.14 (m, 2H), 2.06-1.88 (m, 1H), 1.12 (dt, J= 10.0, 5.5 Hz, 1H), 0.67 (td, J= 5.3, 2.4 Hz, 1H). MS (ES+): 536.1 (M+l); (ES-): 534.1 (M-l); Analysis calculated for C₂4H₂₂BrN₇O₃.2H₂O.lHCl: C, 47.34; H, 4.47; Cl, 5.82; N, 16.10; Found: C, 47.34; H, 4.40;

Cl, 5.48; N, 15.94.

Scheme 55

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (55a)

167

Compound 55a was prepared according to the procedure reported in step-3 of scheme-1, from

TFA sait of 2-(4-amino-8-methyl-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (30e) (50 mg, 0.135 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (44.9 mg, 0.135 mmol), HATU (77 mg, 0.203 mmol), DIPEA (0.118 mL, 0.675 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (55a) (47 mg, 65% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-î7₆) δ 10.75 (s, 1H, D₂O exchangeable), 8.51 (s, 1H), 8.35 (s, 2H, D₂O exchangeable), 8.24 (dd, J= 6.2, 3.0 Hz, 1H), 7.94 (d, J= 8.2 Hz, 1H), 7.64 (t, J= 8.0 Hz, 1H), 7.27 - 7.14 (m, 3H), 5.81 (d, J= 18.0 Hz, 1H), 5.54 (d,J= 17.9 Hz, 1H), 4.31 (dd,J=9.0, 6.1Hz, 1H), 3.63 (dd, J= 5.6, 2.3 Hz, 1H), 2.62 (s, 3H), 2.48-2.45 (m, 1H), 1.91 (dd, J= 13.1, 6.1 Hz, 1H), 1.24 (s, 3H), 0.95 (t, J= 5.4 Hz, 1H), 0.77 (dd, J= 5.4, 2.4 Hz, 1H); MS (ES+): 534.1 (M+l); (ES-): 532.1 (M-l); Analysis calculated for C₂₅H₂₄BrN₇O₂ 1.5H₂O.0.9HCl: C, 50.53; H, 4.73; Cl, 5.37; N, 16.50; Found: C, 50.38; H, 4.74; Cl, 5.68; N, 16.43.

Scheme 56

^,BuO₂C^'Br

CS2CO3

56e 56f

Préparation of (lR,3S,5R)-2-(2-(4-amino-5-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo [3.1.0]hexane-3 -carboxamide (56g)

168

Step-1: Préparation of 2,2,2-trifluoro-N-(2-iodo-3-methylphenyl)acetamide (56b)

Compound 56b was prepared according to the procedure reported in step-1 of scheme-46, from 2-iodo-3-methylaniline (56a) (5 g, 21.45 mmol; CAS # 89938-16-9) in DCM (75 mL) using triethylamine (5.08 mL, 36.5 mmol), trifluoroacetic acid anhydride (4.47 mL, 32.2 mmol) and stirring at RT for 16 h. This gave after workup 2,2,2-trifluoro-N-(2-iodo-3methylphenyl)acetamide (56b) (6.9 g, 21% yield) as a pale yellow solid and was used as such for the next step; Ή NMR (300 MHz, DMSO-cfe) δ 11.26 (s, 1H), 7.43 - 7.24 (m, 2H), 7.17 (dd, 7= 6.1, 3.3 Hz, 1H), 2.47 (s, 3H); MS (ES-): 327.90 (M-l).

Step-2: Préparation of 2-amino-4-methyl-lH-indole-3-carbonitrile (56c)

Compound 56c was prepared according to the procedure reported in step-1 of scheme-11, from 2,2,2-trifluoro-N-(2-iodo-3-methylphenyl)acetamide (56b) (4.9 g, 14.89 mmol) in DMSO (15 mL) using malononitrile (1.125 mL, 17.87 mmol), L-proline (0.343 g, 2.98 mmol), Cul (0.284 g, 1.489 mmol) K2CO3 (4.12 g, 29.8 mmol) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [S1O2 gel (80 g), eluting with EtOAc in hexane from 0-50%] 2-amino-4-methyl-lH-indole-3-carbonitrile (56c) (0.81 g, 32% yield) as a brown solid; Ή NMR (300 MHz, DMSO-7₆) δ 10.67 (s, 1H), 6.96 (d, J= 7.8 Hz, 1H), 6.79 (t, 7= 7.6 Hz, 1H), 6.70 (dt, 7= 7.3, 1.0 Hz, 1H), 6.62 (s, 2H), 2.48 (s, 3H); MS (ES-): 170.10 (M-l).

Step-3: Préparation of 5-methyl-9H-pyrimido[4,5-b]indol-4-amine (56d)

Compound 56d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-4-methyl-lH-indole-3-carbonitrile (56c) (0.8 g, 4.67 mmol) in éthanol (30 mL) using formamidine acetate (3.93 g, 37.4 mmol) and refluxing for 22 h. This gave after workup and purification by reverse-phase column chromatography [Cl8 column (275 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] to afford 5-methyl-9H25 pyrimido[4,5-b]indol-4-amine (56d) (114 mg, 12% yield) HCl sait as a pale yellow solid; ’H NMR (300 MHz, DMSO-7₆) δ 13.15 (s, 1H), 8.65 (s, 1H), 8.03 (s, 2H), 7.49 - 7.32 (m, 2H), 7.22 - 7.05 (m, 1H), 2.94 (s, 3H); MS (ES+): 199.10 (M+l).

Step-4: Préparation of tert-butyl 2-(4-amino-5-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (56e)

Compound 56e was prepared according to the procedure reported in step-2 of scheme-16, from HCl sait of 5-methyl-9H-pyrimido[4,5-b]indol-4-amine (56d) (110 mg, 0.469 mmol) in DMF (3 mL) using tert-butyl 2-bromoacetate (0.076 mL, 0.516 mmol) CS2CO3 (336 mg,

169

1.031 mmol) and stirring at RT for 1.5 h. This gave after workup tert-butyl 2-(4-amino-5methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (56e) (130 mg, 89% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.27 (s, 1H), 7.42 - 7.24 (m, 2H), 7.04 (d, J= 7.1 Hz, 1H), 6.79 (s, 2H), 5.11 (s, 2H), 2.96 (s, 3H), 1.41 (s, 9H); MS (ES+): 313.20 (M+l).

Step-5: Préparation of 2-(4-amino-5-methyl-9H-pyrimido[4,5-b]indol-9-yI)acetic acid (56f)

Compound 56f was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-5-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (56e) (100 mg, 0.320 mmol) using 20% TFA in DCM (1837 pL, 4.80 mmol)) and stirring at RT for 16 h. This gave after workup 2-(4-amino-5-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (56f) (115 mg, 97% yield) TFA sait as a white solid; MS (ES+): 257.10 (M+l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-5-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (56g)

Compound 56g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-5-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (56f) (50 mg,

0.135 mmol) in DMF (1.5 mL) using TFA sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicycIo[3.1.0]hexane-3-carboxamide (4a) (53.5 mg, 0.135 mmol), HATU (77 mg, 0.203 mmol), DIPEA (0.118 mL, 0.675 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography

[C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-5-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (56g) (34 mg, 48% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-cfc) δ 10.76 (s, 1H, D2O exchangeable), 8.60 (s, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.79 - 7.65 (m, 3H, 2H D₂O exchangeable), 7.52 - 7.37 (m,

2H), 7.32 (d, J= 7.7 Hz, 1H), 7.17 (d, J= 7.0 Hz, 1H), 5.74 (d, J= 17.3 Hz, 1H), 5.41 (d, J=

17.3 Hz, 1H), 4.42 (dd, J= 9.1, 5.4 Hz, 1H), 3.94 - 3.86 (m, 1H), 2.96 (s, 3H), 2.36 - 2.13 (m, 2H), 1.99-1.84 (m, 1H), 1.07 (dt, J= 9.3, 5.3 Hz, 1H), 0.78 (dt, J =5.5, 2.7 Hz, 1H). MS (ES+): 520.1 (M+l); 542.1 (M+Na); (ES-): 518.0 (M-l).

Scheme 57

170

57f

Préparation of (lR,3S,5R)-2-(2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (57g)

Step-1: Préparation of 2,2,2-trifluoro-N-(2-iodo-3-methoxyphenyI)acetamide (57b)

Compound 57b was prepared according to the procedure reported in step-1 of scheme-46, from 2-iodo-3-methoxyaniline (57a) (5 g, 20.08 mmol; CAS # 98991-09-4) in DCM (75 mL) using triethylamine (4.76 mL, 34.1 mmol), trifluoroacetic acid anhydride (4.19 mL, 30.1 mmol) and stirring at RT for 16 h. This gave after workup 2,2,2-trifluoro-N-(2-iodo-3methoxyphenyl)acetamide (57b) (6.8 g, 98% yield) as a dark oil and was used as such for next step; MS (ES-): 343.90 (M-l).

Step-2: Préparation of 2-amino-4-methoxy-lH-indole-3-carbonitrile (57c)

Compound 57c was prepared according to the procedure reported in step-1 of scheme-11, from 2,2,2-trifluoro-N-(2-iodo-3-methoxyphenyl)acetamide (57b) (5.1 g, 14.78 mmol) in DMSO (15 mL) using malononitrile (1.117 mL, 17.74 mmol), L-proline (0.340 g, 2.96 mmol), Cul (0.281 g, 1.478 mmol), K2CO3 (4.12 g, 29.8 mmol) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification by flash column

171 chromatography [S1O2 gel (80 g), eluting with EtOAc in hexane from 0-50%] 2-amino-4methoxy-lH-indole-3-carbonitrile (57c) (1.35 g, 49% yield) as a gray solid; ’H NMR (300 MHz, DMSO-iZe) δ 10.54 (s, 1H), 7.00 (d, J= 8.4 Hz, 1H), 6.75 (d, J= 2.3 Hz, 1H), 6.63 6.56 (m, 3H), 3.71 (s, 3H); MS (ES+): 188.10 (M+l); (ES-): 186.10 (M-l).

Step-3: Préparation of 5-methoxy-9H-pyrimido[4,5-b]indol-4-amine (57d)

Compound 57d was prepared according to the procedure reported in step-2 of scheme-29, 2amino-4-methoxy-lH-indole-3-carbonitrile (57c) (1.3 g, 6.94 mmol) in éthanol (45 mL) using formamidine acetate (5.84 g, 55.6 mmol) and refluxing for 22 h. This gave after workup and purification by reverse phase column chromatography [Cl 8 column (275 g), 10 eluting with ACN in water (containing 0.1% HCl) from 0-100%] 5-methoxy-9Hpyrimido[4,5-b]indol-4-amine (57d) (0.9 g, 61% yield) HCl sait as a pale yellow solid; 'H NMR (300 MHz, DMSO-cfc) δ 12.97 (s, 1H), 8.60 (s, 2H), 8.55 (s, 1H), 8.40 (d, J= 8.7 Hz, 1H), 7.09 (d, J= 2.3 Hz, 1H), 7.03 (dd, J= 8.7, 2.4 Hz, 1H), 3.87 (s, 3H); MS (ES+): 215.10 (M+l);

Step-4: Préparation of ZerZ-butyl 2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetate (57e)

Compound 57e was prepared according to the procedure reported in step-2 of scheme-16, from HCl sait of 5-methoxy-9H-pyrimido[4,5-b]indol-4-amine (57d) (810 mg, 3.23 mmol) in DMF (20 mL) using ZerZ-butyl 2-bromoacetate (0.525 mL, 3.55 mmol), CS2CO3 (2316 mg, 20 7.11 mmol) and stirring at RT for 1.5 h. This gave after workup ZerZ-butyl 2-(4-amino-5methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (57e) (990 mg, 93% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.31 - 8.07 (m, 2H), 7.18 - 7.10 (m, 3H), 6.88 (dd, J = 8.6, 2.3 Hz, 1H), 5.10 (s, 2H), 3.84 (s, 3H), 1.41 (s, 9H); MS (ES+): 329.20 (M+l).

Step-5: Préparation of 2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (57f)

Compound 57f was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (57e) (200 mg, 0.609 mmol) using 20% TFA in DCM (3496 pL, 9.14 mmol)) and stirring at RT for 16 h. This gave after workup 2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (57f) (225 mg, 96% yield) TFA sait as a white solid; 'H NMR (300 MHz, DMSO-flfe) δ 8.53 (s, 1H), 8.45 30 8.34 (m, 2H), 7.44 (d, J= 2.2 Hz, 1H), 7.05 (dd, J= 8.7, 2.2 Hz, 1H), 5.26 (s, 2H), 3.88 (s,

3H); MS (ES+): 273.10 (M+l).

172

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (57g)

Compound 57g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (57f) (50 mg,

0.129 mmol) in DMF (1.5 mL) using TFA sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (51.3 mg, 0.129 mmol), HATU (73.8 mg, 0.194 mmol), DIPEA (0.113 mL, 0.647 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography

[Cl 8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (57g) (35 mg, 50% yield) HCl sait as a white solid; ¹H NMR (300 MHz, DMSO-îZé) δ 10.74 (s, 1H, D2O exchangeable), 8.53 (s, 1H), 8.42 - 8.19 (m, 3H, 2H D₂O exchangeable), 8.02 (d, J= 8.2 Hz, 1H), 7.71 (t, J=

8.0 Hz, 1H), 7.33 (d, J= 7.7 Hz, 1H), 7.25 (d, J= 2.3 Hz, 1H), 7.04 (dd, 8.7, 2.2 Hz, 1H),

5.71 (d, J= 17.2 Hz, 1H), 5.42 (d, J= 17.2 Hz, 1H), 4.42 (dd, J= 9.0, 5.6 Hz, 1H), 3.97 3.86 (m, 4H), 2.28 (m, 2H), 1.99 - 1.86 (m, 1H), 1.08 (m, 1H), 0.81 - 0.70 (m, 1H). MS (ES+): 536.1 (M+l); 558.1 (M+Na); (ES-): 534.1 (M-l); Analysis calculated for

C24H₂₂BrN7O3 1.5H₂O.1HC1: C, 48.05; H, 4.37; Cl, 5.91; N, 16.34; Found: C, 48.01; H, 4.33;

Cl, 5.97; N, 16.16.

Scheme 58

58c

Préparation of (lR,3S,5R)-2-(2-(6-acetyl-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (58c)

Step-1: Préparation of ethyl 2-(6-acetyl-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetate (58a)

To a suspension of ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (200 mg, 0.573 mmol) in toluene (8 mL) was added tributyl(l-ethoxyvinyl)stannane (0.279 mL, 0.802 mmol) and the mixture was degassed and filled with nitrogen followed by addition of Pd(PPh3)₄ (132 mg, 0.115 mmol) and heating for 14 h at 120 °C. Reaction mixture was cooled to RT, diluted with ethyl acetate (150 mL), treated with IN HCl (30 mL), water (30 mL) followed by stirring at RT for 15 min. The mixture was treated with 2 M K2CO3, extracted, and separated. The organic layer was washed with brine (60 mL), dried, fîltered and concentrated in vacuum. The residue obtained was purified by flash column chromatograph [silica gel (24 g), eluting with methanol in DCM (0-5%)] to afford ethyl 2-(6acetyl-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetate (58a) (20 mg, 11% yield) as a white solid; 1H NMR (300 MHz, DMSO-t/g) δ 8.9g (d, J= 1. g Hz, 1H), 8.34 (s, 1H), 8.01 (dd, J= 8. g, 1. g Hz, 1H), 7.72 (d, J= 8.7 Hz, 1H), 7. gO (s, 2H), g. 29 (s, 2H), 4.14 (q, J= 7.1 Hz, 2H), 2.71 (s, 3H), 1.20 (t, J= 7.1 Hz, 3H) ; MS (ES+): 313.10 (M+l).

Step-2: Préparation of 2-(6-acetyl-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (58b)

174

Compound 58b was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(6-acetyl-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetate (58a) (18 mg, 0.058 mmol) in THF (3 mL) and methanol (3 mL) using a solution of lithium hydroxide hydrate (14.81 mg, 0.346 mmol) in water (3 mL) and stirring at RT for 14 h. This gave after workup

2-(6-acetyl-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (58b) (17 mg, 100 %) which was used as such in next step-3 without further purification; MS (ES+): 285.10 (M+l), (ES-): 283.0 (M-l)

Step-3: Préparation of (lR,3S,5R)-2-(2-(6-acetyl-4-amino-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (58c)

Compound 58c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(6-acetyl-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (58b) (17 mg, 0.058 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (74 mg, 0.232 mmol), HATU (88 mg, 0.232 mmol), DIPEA (0.061 mL, 0.348 mmol) and stirring at RT for 20 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by conversion of product obtained to HCl sait by dissolving in acetonitrile (2 mL) and 0.1% aq. HCI (15 mL) and lyophilization (lR,3S,5R)-2-(2-(6-acetyl4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (58c) (27 mg, 85% yield) HCl sait as an off-white solid; ’H NMR (300 MHz, DMSO-Æ) δ 10.76 (s, 1H, D₂O exchangeable), 9.09 (d, J= 1.6 Hz, 1H), 8.73 - 8.50 (m, 3H, 2H D₂O exchangeable), 8.11 (dd, J= 8.7, 1.6 Hz, 1H), 8.00 (d, J= 8.2 Hz, 1H), 7.77 (d, J= 8.7 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.79 (d, J= 17.3 Hz, 1H), 5.45 (d, J= 17.3 Hz, 1H), 4.41 (dd, J= 9.0, 5.6 Hz, 1H), 4.00 3.84 (m, 1H), 2.72 (s, 3H), 2.39-2.12 (m, 2H), 1.99-1.82 (m, 1H), 1.14-1.01 (m, 1H),

0.89 - 0.71 (m, 1H); MS (ES+): 548.10 & 550.10 (M+l); MS (ES-): 546.00 & 548.10 (M-l).

Scheme 59

175

Préparation of (ÎR,3S,5R)-2-(2-(4-amino-6-vinyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (59c)

Step-1: Préparation of ethyi 2-(4-amino-6-vinyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (59a)

To a degassed solution of ethyi 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (200 mg, 0.573 mmol) in dioxane (8 mL) was added potassium vinyltrifluoroborate (153 mg, 1.146 mmol), Pd(PPh3)4 (132 mg, 0.115 mmol), a solution of K2CO3 (158 mg, 1.146 mmol) in water (2.3 mL) and heated at 80 °C for 20 h under nitrogen. The reaction mixture was diluted with ethyi acetate (100 mL), washed with water (50 mL), brine (50 mL), 10 dried, filtered and concentrated in vacuum. The residue obtained was purifïed by flash column chromatography [silica gel (25 g) eluting with 10% methanol in ethyi acetate in hexanes from 0 - 50%] to afford ethyi 2-(4-amino-6-vinyl-9H-pyrimido[4,5-b]indol-9yl)acetate (59a) (82 mg, 48% yield) as a light yellow solid; *H NMR (300 MHz, DMSO-î/6) δ 8.49 (s, 1H), 8.28 (s, 1H), 7.56 (d, J= 8.5 Hz, 1H), 7.50 (dd, J= 8.8, 1.5 Hz, 1H), 7.38 (s,

2H), 6.84 (dd, J= 17.7, 11.0 Hz, 1H), 6.03 - 5.95 (m, 1H), 5.29-5.16 (m, 3H), 4.14 (q, J=

7.1 Hz, 2H), 1.20 (t, J= 7.1 Hz, 3H); MS (ES+): 297.10 (M+l).

Step-2: Préparation of 2-(4-amino-6-vinyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (59b)

Compound 59b was prepared according to the procedure reported in step-4 of scheme-17, from ethyi 2-(4-amino-6-vinyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (59a) (80 mg, 0.270

176 mmol) in THF (3 mL) and methanol (3 mL) using a solution of lithium hydroxide hydrate (69.4 mg, 1.620 mmol) in water (3 mL) and stirring at RT for 20 h. This gave after workup 2(4-amino-6-vinyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (59b) (72 mg, 100 %) which was used as such in next step-3 without further purification; MS (ES+): 269.10 (M+l), (ES-):

267.0 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-vinyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (59c)

Compound 59c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-vinyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (59b) (72 mg, 0.27 mmol) in

DMF (12 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicycIo[3.1.0]hexane-3-carboxamide (4a) (172 mg, 0.540 mmol), HATU (308 mg, 0.810 mmol) DIPEA (0.235 mL, 1.350 mmol) and stirring at RT for 20 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0 - 5%] followed by conversion of product obtained to HCl sait by dissolving in acetonitrile (5 mL) and 0.1% aq. HCl (30 mL) and lyophilization (lR,3S,5R)-2-(2-(4-amino6-vinyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (59c) (54 mg, 38% yield) HCl sait as a light brown solid; 'HNMR (300 MHz, DMSO-J₆) δ 10.76 (s, 1H, D₂O exchangeable), 8.60 (s, 1H), 8.54 (s, 1H), 8.36 (s, 2H, D₂O exchangeable), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H),

7.63 (s, 2H), 7.32 (d, J= 7.7 Hz, 1H), 6.86 (dd, J= 17.8, 10.8 Hz, 1H), 6.01 (d, J= 17.7 Hz,

1H), 5.72 (d, J= 17.5 Hz, 1H), 5.39 (d, 17.1 Hz, 1H), 5.29 (d, J= 11.0 Hz, 1H), 4.47 4.36 (m, 1H), 3.96-3.85 (m, 1H), 2.43-2.13 (m, 2H), 2.00-1.83 (m, 1H), 1.14-0.99 (m, 1H), 0.85-0.67 (m, 1H); MS (ES+): 532.10 & 534.10 (M+l); MS (ES-): 530.05 & 532.10 (M-l); Analysis calculated for C₂5H₂₂BrN₇O₂.1.0HC1.2.25H₂O: C, 49.27; H, 4.55; Cl, 5.82;

N, 16.09; Found: C, 49.02; H, 4.49; Cl, 5.61; N, 16.04.

Scheme 60

177

49a 60a

^60b 60c

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxamide (60c)

Step-1: Préparation of ethyl 2-(4-amino-6-carbamoyl-9H-pyrimido[4,5-b]indol-9-yl)acetate 5 (60a)

A suspension of ethyl 2-(4-amino-6-cyano-9H-pyrimido[4,5-b]indol-9-yl)acetate (49a) (70 mg, 0.237 mmol) in éthanol (6 mL) was treated with conc. NH4OH (2.25 mL) followed by hydrogen peroxide (0.084 mL, 0.948 mmol) and stirred at RT for 48 h. The solid obtained was collected by filtration, washed with éthanol and dried to obtain ethyl 2-(4-amino-610 carbamoyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (60a) (40 mg, 54%) as a white solid residue which was used as such for next step; MS (ES+): 314.10 (M+l).

Step-2: Préparation of 2-(4-amino-6-carbamoyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (60b)

Compound 60b was prepared according to the procedure reported in step-4 of scheme-17, 15 from ethyl 2-(4-amino-6-carbamoyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (60a) (38 mg, 0.121 mmol) in THF (2 mL) and methanol (2 mL) using a solution of lithium hydroxide hydrate (31.2 mg, 0.728 mmol) in water (2 mL) and stirring at RT for 68 h. This gave after workup 2-(4-amino-6-carbamoyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (60b) (35 mg,

178

100 %) which was used as such in next step-3 without further purification; MS (ES+): 286.10 (M+l), (ES-): 284.0 (M-l).

Step-3: Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxamide (60c)

Compound 60c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-carbamoyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (60b) (34.5 mg, 0.121 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (77 mg, 0.242 mmol), HATU (92 mg, 0.242 mmol) DIPEA (0.105 mL, 0.605 mmol) and stirring at RT for 19 h. This gave after workup and purification by flash column chromatography [silica gel (4 g), eluting with MeOH in DCM from 0 - 10%] followed by purification by reverse phase column chromatography [Cl8 column, eluting with ACN in water (containing 0.1% HCl) from 0-100%] of 4-amino-9-(2((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxamide (60c) (12 mg, 18% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-cfe) δ 10.76 (s, 1H), 9.03 (s, 1H), 8.57 (s, 1H), 8.36 (s, 2H, D₂O exchangeable), 8.08 - 7.97 (m, 2H), 7.89 (s, 1H, D2O exchangeable), 7.78 - 7.63 (m, 2H), 7.47 (s, 1H, D2O exchangeable), 7.31 (d, J= 7.7 Hz, 1H), 5.76 (d, J= 17.3 Hz, 1H), 5.42 (d, J= 17.4 Hz, 1H), 4.42 (dd, J= 9.2, 5.5 Hz, 1H), 3.95 - 3.86 (m, 1H), 2.41 - 2.10 (m, 2H), 2.01 - 1.74 (m, 1H), 1.18 - 0.98 (m, 1H), 0.87 - 0.69 (m, 1H); MS (ES+): 549.10 &

551.10 (M+l); MS (ES-): 547.05 & 549.10 (M-l).

Scheme 61

^48d 61a

Préparation of (lR,3S,5R)-N-(6-bromopyridin-2-yI)-2-(2-(4,6-diamino-9H-pyrimido[4,5b] indol-9-yI)acetyl)-2-azabicyclo [3.1.0]hexane-3 -carboxamide (61 a)

179

Compound 61a was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl (4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indoI-6-yl)carbamate (48d) (6 mg, 9.65 pmol) in DCM (5 mL) using TFA. This gave after workup and purification by reverse phase column chromatography [Cl8 column, eluting with ACN in water (containing 0.1% HCl) from 0-100%](lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-(2-(4,6-diamino-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (61a) (3.9 mg, 77% yield) HCl sait as a white solid;. Ή NMR (300 MHz, DMSO-îZ₆) δ 10.76 (s, 1H), 8.47 (s, 1H), 8.28 (s, 1H), 8.09 (s, 2H), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.1 Hz, 1H), 7.64 (d, J = 8.8 Hz, 1H), 7.46 - 7.35 (m, 1H), 7.32 (d, J= 7.6 Hz, 1H), 5.69 (d, J= 17.4 Hz, 1H), 5.37 (d, J= 17.4 Hz, 1H), 4.48 - 4.34 (m, 1H), 3.97 - 3.83 (m, 1H), 2.39 - 2.09 (m, 2H), 2.05 1.73 (m, 1H), 1.17-0.99 (m, 1H), 0.84-0.67 (m, 1H); MS (ES+): 521.10 & 523.10 (M+l).

Scheme 62

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo [3.1.0]hexane-3 -carboxamide (62c)

Step-1: Préparation of ethyl 2-(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (62a)

A degassed solution of ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (150 mg, 0.43 mmol) and phenylboronic acid (79 mg, 0.644 mmol) in dioxane (15 mL)

180

was treated with césium carbonate (210 mg, 0.644 mmol) in water (1.8 mL), Pd(PPh3)2Cb (60.3 mg, 0.086 mmol) and heated at 100 °C for 14 h under nitrogen. The reaction mixture was diluted with ethyl acetate/methanol (2:1, 75 mL), filtered, washed with ethyl acetate/methanol (2:1), and the filtrate was concentrated in vacuum. The residue obtained was 5 purifïed by flash column chromatography [S1O2 gel (25 g), eluting with MeOH in DCM from

5-40%] to afford ethyl 2-(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (62a) (86 mg, 58% yield) as an off-white solid; MS (ES+): 347.15.

Step-2: Préparation of 2-(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (62b)

Compound 62b was prepared according to the procedure reported in step-4 of scheme-17, 10 from ethyl 2-(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (62a) (80 mg, 0.231 mmol) in THF (2 mL) and methanol (2 mL) using a solution of lithium hydroxide hydrate (59.3 mg, 1.386 mmol) in water (2 mL) and stirring at RT for 19 h. This gave after workup 2(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (62b) (59 mg, 80 %) which was used as such in next step-3 without further purification; MS (ES+): 319.10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (62c)

Compound 62c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (62b) (59 mg, 0.185 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-220 azabicyclo[3.1.0]hexane-3-carboxamide (4a) (89 mg, 0.278 mmol), HATU (141 mg, 0.371 mmol) DIPEA (0.097 mL, 0.556 mmol) and stirring at RT for 21 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with hexanes/10% MeOH in EtOAc in hexanes from 0 -100%] followed by conversion of product obtained to HCl sait by dissolving in acetonitrile (2 mL) and 0.1% aq. HCl (15 mL) and lyophilization (lR,3S,5R)-2-(2-(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (62c) (51.5 mg, 48% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 10.77 (s, 1H, D₂O exchangeable), 8.77 (s, 1H), 8.56 (s, 1H), 8.43 (s, 2H, D2O exchangeable), 8.01 (d, J= 8.1 Hz, 1H), 7.90 - 7.80 (m, 3H), 7.79 - 7.66 (m, 2H), 7.55 - 7.47 (m, 2H), 7.42 - 7.36 (m, 1H), 30 7.32 (d, J= 7.7 Hz, 1H), 5.77 (d, J= 17.3 Hz, 1H), 5.44 (d, J= 17.3 Hz, 1H), 4.42 (dd, J=

9.0, 5.5 Hz, 1H), 3.99 - 3.86 (m, 1H), 2.42 - 2.15 (m, 2H), 2.00 - 1.79 (m, 1H), 1.16 - 0.99

181 (m, 1H), 0.85 - 0.70 (m, 1H); MS (ES+): 582.10 & 584.20 (M+l); MS (ES-): 580.10 & 582.10 (M-l); Analysis calculated for C₂₉H₂4BrN₇O₂.1.0HC1.1.5H₂O: C, 53.92; H, 4.37; N, 15.18; Cl, 5.49; Found: C, 54.10; H, 4.48; N, 14.92; Cl, 5.28.

h₂n^^_n/^nh₂ h

NiCI₂, 6H₂O, NaBH₄

Scheme 63

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(aminomethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (63d)

Step-1: Préparation of ethyl/methyl 2-(4-amino-6-((ter/-biitoxycarbonylamino)methyl)-9Hpyrimido[4,5-b]indol-9-yl)acetate (63a)

To a solution of ethyl 2-(4-amino-6-cyano-9H-pyrimido[4,5-b]indol-9-yl)acetate (49a) (100 mg, 0.339 mmol) in MeOH (8 mL) cooled with ice/water and was added di-tertebutyl dicarbonate (299 mg, 1.355 mmol), nickel(II) chloride hexahydrate (40.2 mg, 0.169 mmol) followed by sodium borohydride (78 mg, 2.032 mmol) slowly over a period of 5 min and was stirred at RT for 1 h. The reaction mixture was treated with Nl-(2-aminoethyl)ethane-l,215 diamine (0.148 mL, 1.355 mmol) and stirred for 30 min at RT and concentrated to diyness. The residue obtained was purified by flash column chromatography [silica gel (25 g), with 10% methanol in ethyl acetate in hexanes/ from 0-100%, followed by methanol in dichloromethane from 0-33% and DMA80] to afford a mixture of ethyl/methyl 2-(4-amino-6((tert-butoxycarbonylamino)methyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (63a) (7 mg, 5%)

182 as a colorless gum; MS (ES+): 400.20 (M+l); 386.20 (M+l) and 2-(4-amino-6-(((ZerZbutoxycarbonyl)amino)methyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (63b) (15 mg, 12%) as a colorless gum; MS (ES+): 372.20 (M+l), (ES-): 370.15 (M-l).

Step-2: Préparation of 2-(4-amino-6-(((ZerZ-butoxycarbonyl)amino)methyl)-9H-pyrimido[4,55 b]indol-9-yl)acetic acid (63b)

Compound 63b was prepared according to the procedure reported in step-4 of scheme-17, from a mixture of ethyl/methyl 2-(4-amino-6-((terZ-butoxycarbonylamino)methyl)-9Hpyrimido[4,5-b]indol-9-yl)acetate (63a) (7 mg, 0.018 mmol) in THF (2 mL) and methanol (2 mL) using a solution of lithium hydroxide hydrate (6 mg, 0.140 mmol) in water (2 mL) and stirring at RT for 19 h. This gave after workup 2-(4-amino-6-(((ZerZbutoxycarbonyl)amino)methyl)-9H-pyrimido[4,5-b]indol-9-yl)açetic acid (63b) which was used as such in next step-3 without further purification; MS (ES+): 372.20 (M+l), (ES-): 370.10 (M-l).

Step-3: Préparation of ZerZ-butyl ((4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-215 yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyI)-9H-pyrimido[4,5-b]indol-6yl)methyl)carbamate (63c)

Compound 63c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(((Zeri-butoxycarbonyl)amino)methyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (63b) (21 mg, 0.057 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(620 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (55 mg, 0.171 mmol), HATU (65 mg, 0.171 mmol), DIPEA (0.05 mL, 0.285 mmol) and stirring at RT for 19 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), 10% MeOH in EtOAc in hexane from 0 - 100%] ZerZ-butyl ((4-amino-9-(2-((lR,3S,5R)-3((6-bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyI)-9H25 pyrimido[4,5-b]indol-6-yl)methyl)carbamate (63c) (8 mg) as an off-white solid; MS (ES+): 635.20 & 637.20 (M+l), (ES-): 633.10 & 635.10 (M-l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(aminomethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromppyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (63d)

Compound 63d was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl ((4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6-yl)methyl)carbamate

183 (63c) (8 mg, 0.013 mmol) in DCM (5 mL) using TFA (0.097 mL, 1.259 mmol) and stirring at RT for 21 h. This gave after workup and purification by reverse phase column chromatography [Cl8 column, eluting with ACN in water (containing 0.1% HCl) from 0100%](lR,3S,5R)-2-(2-(4-amino-6-(aminomethyl)-9H-pyrimido[4,5-b]indoI-9-yl)acetyl)-N5 (6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (63d) (1.6 mg, 24% yield)

HCl sait as a white solid; 'H NMR (300 MHz, MethanoW₄) δ 8.56 (s, 1H), 8.51 (d, J= 1.5 Hz, 1H), 8.05 (d, J= 8.2 Hz, 1H), 7.86 (d, J= 8.5 Hz, 1H), 7.72 (dd, J= 8.6, 1.7 Hz, 1H), 7.61 (t, J= 8.0 Hz, 1H), 7.26 (dd, J= Ί.Ί, 0.7 Hz, 1H), 5.84 - 5.61 (m, 2H), 4.58 - 4.45 (m, lH),4.33(s, 2H), 3.98-3.88 (m, 1H), 2.57-2.30 (m, 2H), 2.13-1.95 (m, 1H), 1.29-1.14 (m, 1H), 0.95 - 0.81 (m, 1H); MS (ES+): 535.10 & 537.10 (M+l).

Scheme 64

5c

Préparation of (2S,4R)-l-(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (64a)

Compound 64a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (5c) (158 mg, 0.515 mmol) in DMF (5 mL) using TFA sait of (2S,4R)-N-(3-chloro-2-fluorobenzyI)-4fluoropyrrolidine-2-carboxamide (7a) (200 mg, 0.515 mmol), HATU (235 mg, 0.617 mmol) DIPEA (0.449 mL, 2.57 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-1(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (64a) (115 mg, 50% yield) HCl sait as a white solid; *H

184

NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 9.28 (bs, 1H, D2O exchangeable), 9.14 and 8.70 (2t, J= 5.9 Hz, 1H), 8.54 (bs, 1H, D2O exchangeable), 8.35 and 8.34 (2s, 1H), 7.55 - 7.34 (m, 2H), 7.21 (dt, J= 22.4, 7.6 Hz, 1H), 7.07 (t, J= 7.9 Hz, 1H), 6.95 (dd, J= 6.0, 3.4 Hz, 1H), 5.62 - 4.55 (m, 3H), 4.39 (td, J= 16.7, 6.9 Hz, 2H), 4.24 (dd, J= 15.9, 5.7 5 Hz, 1H), 4.16 - 3.99 (m, 1H), 3.87 (ddd, J= 37.7, 12.3, 3.0 Hz, 1H), 2.62 - 2.38 (m, 1H), 2.22-1.92 (m, 1H); ^{10 * * * * 15 * * * I9 20 * * * * 25}F NMR (282 MHz, DMSO-7₆) δ -121.32, -121.65, -176.12, -176.32; MS (ES+): 449/451 (M+l).

Préparation of (2S,4R)-l-(2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (65a)

Compound 65a was prepared according to the procedure reported in step-3 of scheme-1, from

TFA sait of 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetic acid (2c) (150 mg, 0.49 mmol) in DMF (5 mL) using TFA sait of (2S,4R)-N-(3-chIoro-2-fluorobenzyl)-4-fluoropyrrolidine15 2-carboxamide (7a) (190 mg, 0.490 mmol), HATU ((224 mg, 0.588 mmol) DIPEA (0.428 mL, 2.449 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-1-(2-(420 aminopyrrolo[2,1 -f] [ 1,2,4]triazin-7 -y l)acetyl)-N-(3 -chloro-2-fluorobenzy l)-4fluoropyrrolidine-2-carboxamide (65a) (104 mg, 47.3 % yield) HCl sait as a white solid; *H

NMR (300 MHz, DMSO-Jô) (a mixture of two rotamers) δ 9.81 (bs, 1H, D₂O exchangeable),

9.21 - 8.91 (m, 1H, D₂O exchangeable), 8.65 (t, 7 = 5.9 Hz, 1H), 8.14 and 8.12 (2s, 1H), 7.54

-7.38 (m, 2H), 7.38-7.26 (m, 1H), 7.14 (dt, 7= 15.6, 7.9 Hz, 1H), 6.78 and 6.66 (2d,7=

4.5 Hz, 1H), 5.56 - 5.20 (m, 1H), 4.51 - 4.22 (m, 3H), 4.18 - 3.34 (m, 4H), 2.63 - 2.38 (m,

1H), 2.19-1.88 (m, 1H); ^I9F NMR (282 MHz, DMSO-7₆) δ -121.27, -121.68, -176.19, 21436

185

176.45; MS (ES+) 449/451 (M+l); Analysis calculated for C20H19CIF2N6O2.HCI.I.75H2O: C, 46.48; H, 4.58; Cl, 13.72; N, 16.26 Found: C, 46.64; H, 4.31; Cl, 13.61; N, 15.94.

Scheme 66

Préparation of (lR,3S,5R)-2-(2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetyl)-N-(3chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (66b)

Compound 66b was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetic acid (2c) (100 mg, 0.327 mmol) in DMF (5 mL) using TFA sait of (lR,3S,5R)-N-(3-chloro-2-fluorobenzyl)-210 azabicyclo[3.1.0]hexane-3-carboxamide (66a) (125 mg, 0.327 mmol; prepared according to the procedure reported by Wiles, Jason A. et al. PCT Int. Appl. (2017), WO 2017035349 Al 20170302), HATU (149 mg, 0.392 mmol) DIPEA (0.285 mL, 1.633 mmol) and stirring atRT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%](lR,3S,5R)-2-(2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetyl)N-(3-chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (66b) (59 mg, 40.8 % yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-Jâ) (a mixture of two rotamers) δ 9.93 (s, 1H, D2O exchangeable), 9.14 (s, 1H, D2O exchangeable), 8.46 (t, J= 5.9 Hz, 1H,

D₂O exchangeable), 8.17 (s, 1H), 7.55 - 7.41 (m, 2H), 7.25 (t, 1H), 7.14 (t, J= 7.8 Hz, 1H), 6.82 (d, J= 4.6 Hz, 1H), 4.42 - 4.12 (m, 5H), 3.67 - 3.55 (m, 1H), 2.31 - 2.05 (m, 2H), 1.90 - 1.70 (m, 1H), 1.04 - 0.86 (m, 1H), 0.70 - 0.53 (m, 1H); ¹⁹F NMR (282 MHz, DMSOifc) δ -121.74; MS (ES+): 443/445 (M+l); Analysis calculated for C21H20CIFN6O2.HCI.2.5H2O: C, 48.10; H, 5.00; Cl, 13.52; N, 16.03; Found: C, 48.08; H, 4.82; Cl, 13.96; N, 15.88.

Scheme 67

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (67c)

Compound 67c was prepared according to the procedure reported in step-3 of scheme-1, from 5 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (65 mg, 0.337 mmol) in DMF (1.5 mL) and

DMSO (0.5 mL) using (lR,3S,5R)-N-(3-chloro-2-fluorobenzyl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (67b) (114 mg, 0.404 mmol), HATU (96 mg, 0.252 mmol), DIPEA (0.293 mL, 1.683 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in 10 DCM from 0 - 100%] followed by purification using reverse phase column chromatography

[Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (67c) (92mg, 60% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.52 (t, J= 5.9 Hz, 1H, D₂O exchangeable), 8.39 (s, 1H), 15 8.33 (s, 1 H), 7.47 (td, J= 7.6, 1.7 Hz, 1H), 7.28 - 7.19 (m, 1H), 7.13 (t, J= 7.9 Hz, 1H), 5.46 (d, J= 17.1 Hz, 1H), 5.20 (d, J= 17.0 Hz, 1H), 4.33 (d, J= 5.8 Hz, 2H), 4.24 (dd, J= 9.2, 4.7 Hz, 1H), 3.46 (dd, J= 5.5, 2.4 Hz, 1H), 2.40 (dd, J= 13.2, 9.2 Hz, 1H), 1.91 (dd, J= 13.2, 4.7 Hz, 1H), 1.25 (s, 3H), 1.00 (t, J= 5.3 Hz, 1H), 0.89 (dd, J= 5.3, 2.4 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -121.57. MS (ES+): 458.1 (M+l), 480.1 (M+Na); (ES-): 456.7 (M20 1), 492.5 (M+Cl); Analysis calculated for C₂iH₂iC1FN₇O₂.HC1.1.75H₂O: C, 47.96; H, 4.89;

Cl, 13.48; N, 18.64; Found: C, 47.91; H, 4.55; Cl, 13.17; N, 18.43.

Scheme 68

Préparation of (2S,4R)-1 -(2-(6-amino-9H-purin-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2yl)-4-fluoropyrrolidine-2-carboxamide (68a)

Compound 68a was prepared according to the procedure reported in step-3 of scheme-1, from 5 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (65 mg, 0.337 mmol) in DMF (1.5 mL) and

DMSO (0.5 mL) using (2S,4R)-N-(6-bromo-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2carboxamide (3a) (102 mg, 0.337 mmol), HATU (192 mg, 0.505 mmol), DIPEA (0.293 mL, 1.683 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(6-amino-9Hpurin-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (68a) (45 mg, 28% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-îZ₆) δ (a mixture of two rotamers) 10.92 (s) and 10.52 (s) (2s, 1H) (D2O exchangeable), 9.50 (s, 1H) (D2O exchangeable), 8.76 (s, 1H) (D2O exchangeable), 8.46 (m, J= 3.0 Hz, 1H), 8.39 (s,

1H), 7.71 (d, J= 8.0 Hz,) and 7.61 (d, J= 8.0 Hz) (2d, 1H), 7.52 (d, J= 7.9 Hz) and 7.43 (d, J= 7.9 Hz) (2d, 1H), 5.71 - 5.17 (m, 3H), 4.60 (t, J= 8.5 Hz, 1H), 4.29 - 3.81 (m, 2H), 2.74 - 2.52 (m, 1H), 2.30 - 2.04 (m, 1H), 2.02 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-7₆) δ -175.88; MS (ES+): 477.1 (M+l), 499.1 (M+Na); (ES-): 475.6 (M-l); Analysis calculated for

Ci8Hi₈BrFN₈O2.1.2HC1.2.5H2O: C, 38.19; H, 4.31; Cl, 7.52; N, 19.79; Found: C: 38.33; H,

3.98; Cl, 7.31; N, 19.64.

Scheme 69

Préparation of (2S,4R)-1 -(2-(4-aminopyrrolo[2,1 -f][ 1,2,4]triazin-7-yl)acetyl)-N-(6bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (69a)

Compound 69a was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetic acid (2c) (75 mg, 0.245 mmol) in DMF (5 mL) using TFA sait of (2S,4R)-N-(6-bromopyridin-2-yl)-4-fluoropyrroIidine-2carboxamide (5d) (98 mg, 0.294 mmol), HATU (112 mg, 0.294 mmol) and DIPEA (158 mg, 1.225 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (2S,4R)-l-(2-(4-aminopyrrolo[2,lf] [ 1,2,4]triazin-7-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (69a) (40 mg, 35 % yield) as a white solid; Ή NMR (300 MHz, DMSO-cfc) δ (a mixture of rotamers) 11.28 (s) and 10.99 (s) (2s, 1H, D₂O exchangeable), 9.86 (s, 1H, D₂O exchangeable), 9.09 (s, 1H, D₂O exchangeable), 8.16 (s, 1H), 8.11 - 7.99 (m, 1H), 7.81 7.70 (m, 1H), 7.51 - 7.39 (m, 1H), 7.35 (d, J= 8.2 Hz, 1H), 6.75 (dd, J= 12.6, 4.6 Hz, 1H), 5.45 (dt, J= 52.6, 2.9 Hz, 1H), 4.64 (t, J= 8.5 Hz, 1H), 4.18 - 3.97 (m, 2H), 3.90 (dd, J= 12.5, 3.1 Hz, 1H), 3.81 - 3.73 (m, 1H), 2.61 - 2.40 (m, 1H), 2.30 - 1.95 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-îZs) δ -175.77, -176.09; MS (ES+): 462.0 (M+l); (ES-): 460.0 (M-l).

Scheme 70

189

nh₂ 2c 70a

Préparation of (lR,3S,5R)-2-(2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (70a)

Compound 70a was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetic acid (2c) (75 mg, 0.245 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (78 mg, 0.245 mmol), HATU (112 mg, 0.294 mmol), DIPEA (158 mg, 1.225 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in

DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (lR,3S,5R)-2-(2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetyl)-N-(6-bromopyridin-2-yl)2-azabicyclo[3.1.0]hexane-3-carboxamide (70a) (64 mg, 57% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-î/₆) δ 10.75 (s, 1H, D₂O exchangeable), 9.91 (s, 1H, D₂O exchangeable), 9.13 (s, 1H, D₂O exchangeable), 8.19 (s, 1H), 8.05 (d, J = 8.1 Hz, 1H), 7.73 (t, J = 8.0 Hz, 1H), 7.48 (d, J = 4.6 Hz, 1H), 7.34 (d, J = 7.7 Hz, 1H), 6.82 (d, J = 4.6 Hz, 1H), 4.43 (dd, J = 9.0, 5.5 Hz, 1H), 4.31 (d, J = 16.9 Hz, 1H), 4.15 (d, J = 16.9 Hz, 1 H), 3.71 - 3.66 (m, 1H), 2.39-2.12 (m,2H), 1.89-1.72 (m, 1H), 1.02-0.90 (m, 1H), 0.66 - 0.52 (m, 1H); MS (ES+): 456.0 (M+l); (ES-) : 454.0 (M-l); Analysis calculated for

Ci₉Hi₈BrN₇O₂.1.25HC1.2.25H₂O: C, 42.07; H, 4.41; Cl, 8.17; N, 18.08; Found: C, 42.33; H,

4.10; Cl, 7.92; N, 17.95.

Scheme 71

190

Préparation of (lR,3S,5R)-2-(2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (71a)

Compound 71a was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetic acid (2c) (75mg, 0.245 mmol) in DMF (2 mL) and using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (8a) (81 mg, 0.245 mmol), HATU (140 mg, 0.367 mmol), DIPEA (0.213 mL, 1.225 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in 10 DCM from 0 - 100%] followed by purification using reverse phase column chromatography

[Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetyl)-N-(6-bromopyridin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (71a) (59 mg, 51% yield) HCl sait as a white solid; ’HNMR (300 MHz, DMSO-î7₆) δ 10.75 (s, 1H, D₂O exchangeable), 9.98 (s, 1H, 15 D₂O exchangeable), 9.18 (s, 1H, D₂O exchangeable), 8.19 (s, 1H), 8.05 (d, J= 8.2 Hz, 1H), 7.73 (t, J= 8.0 Hz, 1H), 7.50 (d, J= 4.6 Hz, 1H), 7.34 (d, J= 7.7 Hz, 1H), 6.81 (d, J= 4.6 Hz, 1H), 4.39 (dd, J= 9.1, 5.7 Hz, 1H), 4.28 (d, J= 17.0 Hz, 1H), 4.10 (d, J= 16.9 Hz, 1H), 3.46 (dd, J= 5.6, 2.3 Hz, 1H), 2.43 (dd, J= 13.2, 9.1 Hz, 1H), 1.96 (dd, J= 13.2, 5.7 Hz, 1H), 1.24 (s, 3H), 0.89 (t, J= 5.4 Hz, 1H), 0.75 (dd, J= 5.4, 2.3 Hz, 1H); MS (ES+): 469.9 20 (M+l); (ES-): 468.0 (M-l); Analysis calculated for C₂₀H₂₀BrN₇O₂.1.5H₂O.1.15HCl: C,

44.54; H, 4.51; Cl, 7.56; N, 18.18; Found: C, 44.79; H, 4.29; Cl, 7.53; N, 18.23.

Scheme 72

Préparation of (lR,3S,5R)-2-(2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetyl)-N-(6bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (72a)

Compound 72a was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(4-aminopyrrolo[2,l-f][l,2,4]triazin-7-yl)acetic acid (2c) (75mg, 0.245 mmol) in DMF (2 mL) and using HCl sait of (lR,3S,5R)-N-(6-bromo-3-methylpyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (10a) (85 mg, 0.245 mmol), HATU (140 mg, 0.367 mmol), DIPEA (0.213 mL, 1.225 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with 10 DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] ( 1 R,3S,5R)-2-(2-(4-aminopyrroIo[2,1 -f] [ 1,2,4]triazin-7-yl)acetyl)-N-(6-bromo-3methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (72a) (68mg, 57.3 % yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 10.29 (s, 1H, D₂O exchangeable), 10.19 (s, 1H, D₂O exchangeable), 9.33 (s, 1H, D₂O exchangeable), 8.21 (s, 1H), 7.64 (d, J= 8.0 Hz, 1H), 7.57 (d, J= 4.6 Hz, 1H), 7.45 (d, J= 7.9 Hz, 1H), 6.83 (d, J = 4.6 Hz, 1H), 4.37 (dd, J= 9.2, 5.2 Hz, 1H), 4.27 (d, J= 17.0 Hz, 1H), 4.13 (d, J= 17.0 Hz, 1H), 3.44 (dd, J= 5.6, 2.4 Hz, 1H), 2.49 - 2.45 (m, 1H), 2.08 (s, 3H), 2.00 (dd, J= 13.3, 5.2 Hz, 1H), 1.26 (s, 3H), 0.91 (t, J= 5.4 Hz, 1H), 0.79 (dd, J= 5.3, 2.4 Hz, 1H); MS (ES+):

484.0 (M+l); (ES-): 482.0 (M-l); Analysis calculated for C₂iH₂₂BrN7O₂.3.5H₂O.1.5HCl: C,

41.89; H, 5.11; N, 16.28; Found: C, 42.02; H, 5.02; N, 15.97.

Scheme 73

Préparation of (lR,3S,5R)-2-(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (73a)

Compound 73a was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (5c) (75mg, 0.245 mmol) in DMF (2 mL) and using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (81 mg, 0.245 mmol), HATU (140 mg, 0.367 mmol), DIPEA (0.213 mL, 1.225 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in

DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7H-pyrroIo[2,3-d]pyrimidin-7-yl)acetyl)-N-(6-bromopyridin-2yI)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (73a) (72mg, 63 % yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/₆) δ 10.81 (s, 1H, D₂O exchangeable), 9.33 (s,

1H, D₂O exchangeable), 8.54 (s, 1H, D₂O exchangeable), 8.38 (s, 1H), 8.04 (d, J= 8.2 Hz,

1H), 7.73 (t, J= 8.0 Hz, 1H), 7.44 (d, J= 3.6 Hz, 1H), 7.34 (d, J= 7.7 Hz, 1H), 6.95 (d, J = 3.6 Hz, 1H), 5.49 (d, J= 17.1 Hz, 1H), 5.19 (d, J= 17.0 Hz, 1H), 4.40 (dd, J= 9.1, 5.8 Hz, 1H), 3.54 (dd, J= 5.6, 2.3 Hz, 1H), 2.46-2.36 (m, 1H), 1.97 (dd, J= 13.2, 5.8 Hz, 1H), 1.28 (s, 3H), 0.97 (t, J= 5.5 Hz, 1H), 0.84 (dd, J= 5.4, 2.4 Hz, 1H); MS (ES+): 470.0 (M+l);

(ES-): 468.0 (M-l); Analysis calculated for C₂₀H₂₀BrN₇O₂.1.75H₂O.l.lHCl: C, 44.32; H,

4.58; Cl, 7.20; N, 18.09; Found: C, 43.99; H, 4.67; Cl, 6.95; N, 17.78.

Scheme 74

Préparation of (lR,3S,5R)-2-(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(6bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (74a)

Compound 74a was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(4-amino-7H-pynOlo[2,3-d]pyrimidin-7-yl)acetic acid (5c) (75mg, 0.245 mmol) in DMF (2 mL) and using HCl sait of (lR,3S,5R)-N-(6-bromo-3-methylpyridin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (10a) (85 mg, 0.245 mmol), HATU (140 mg, 0.367 mmol), DIPEA (0.213 mL, 1.225 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with

DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(6-bromo-3methylpyridin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (74a) (63 mg, 53% yield) HCI sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.32 (s, 1H, D₂O exchangeable), 9.41 (s, 1H, D₂O exchangeable), 8.58 (s, 1H, D₂O exchangeable), 8.38 (s, 1H), 7.64 (d, J= 8.0 Hz, 1H), 7.49 - 7.38 (m, 2H), 6.99 (d, J= 3.5 Hz, 1H), 5.47 (d, J= 17.0 Hz, 1H), 5.20 (d, J= 16.9 Hz, 1H), 4.38 (dd, J= 9.2, 5.1 Hz, 1H), 3.55 - 3.47 (m, 1H), 2.50 2.44 (m, 1H), 2.06 (s, 3H), 2.04 - 1.96 (m, 1H), 1.30 (s, 3H), 1.00 (t, J= 5.4 Hz, 1H), 0.90 (dd, J= 5.3, 2.4 Hz, 1H); MS (ES+): 484.0 (M+l); (ES-): 482.0 (M-l); Analysis calculated for C₂iH₂₂BrN₇O₂.2.5H₂O.1.55HC1.0.25DMSO: C, 42.78; H, 5.01; Cl, 8.81; N, 16.24;

Found: C, 42.52; H, 5.00; Cl, 8.87; N, 15.99.

Scheme 75

Préparation of (S)-N-(6-bromopyridin-2-yI)-1 -(2-(4-(methylamino)-1 H-pyrazolo[3,4d]pyrimidin-l -yl)acetyl)pyrrolidine-2-carboxamide (75d)

Step-1: Préparation of tert-butyl 2-(4-(methylamino)-lH-pyrazolo[3,4-d]pyrimidin-l5 yl)acetate (75b)

To a solution of /eri-butyl 2-(4-chloro-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetate (75a) (0.3 g, 1.116 mmol; prepared according to the procedure reported by Kotian, Pravin L. et al; From PCT Int. AppL, 2017136395, 10 Aug 2017) in THF (4 mL) was added methanamine in methanol (2.79 mL, 5.58 mmol) and stirred at RT for 2 h. The reaction mixture was concentrated and purified by flash column chromatography [silica gel (12 g), eluting with MeOH/EtOAc (9:1) in hexanes from 0 to 100%] to afford ZerZ-butyl 2-(4-(methylamino)-lHpyrazolo[3,4-d]pyrimidin-l-yl)acetate (75b); MS (ES+): 264.4 (M+l), (ES-): 262.3 (M-l). Step-2: Préparation of 2-(4-(methylamino)-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetic acid (75c)

Compound 75c was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-(methylamino)-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetate (75b) using TFA (1.720 mL, 22.33 mmol) in DCM (4 mL) and stirring at RT for 4 days. The reaction mixture was concentrated in vacuum and was suspended in toluene (10 mL) and evaporated. This gave after workup 2-(4-(methylamino)-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetic acid (75c) (0.23 g, 99 % yield) as white solid; Ή NMR (300 MHz, DMSO-ûfc) δ 9.43 (s, 1H), 8.40 (s,

1H), 8.25 (s, 1H), 5.16 (s, 2H), 3.06 (d, J= 4.8 Hz, 3H); MS (ES+): 208.3 (M+l).

Step-3: Préparation of (S)-N-(6-bromopyridin-2-yl)-l-(2-(4-(methylamino)-lH-pyrazolo[3,4d]pyrimidin-1 -yl)acetyl)pyrrolidine-2-carboxamide (75d)

Compound 75d was prepared according to the procedure reported in step-3 of scheme-1, from 25 2-(4-(methylamino)-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetic acid (75c) (0.08 g, 0.386 mmol) in DMF (3 mL) and using (S)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (13a)

195 (0.104 g, 0.386 mmol), HATU (0.161 g, 0.425 mmol), DIPEA (0.101 mL, 0.579 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with CMA-80 in chloroform from 0 - 40%] (S)-N(6-bromopyridin-2-yl)-1 -(2-(4-(methylamino)-1 H-pyrazolo[3,4-d]pyrimidin-1 yl)acetyl)pyrrolidine-2-carboxamide (75d) (60 mg, 34% yield) as a white solid; 1H NMR (300 MHz, DMSO-d6) δ 10.88 (s, 1H), 8.24 (s, 2H), 8.10 - 7.97 (m, 2H), 7.71 (t, J = 7.9 Hz, 1H), 7.32 (d, J = 7.6 Hz, 1H), 5.24 (s, 2H), 4.52 (dd, J = 8.5, 4.1 Hz, 1H), 3.80 - 3.61 (m, 2H), 2.98 (d, J = 4.5 Hz, 3H), 2.24 - 2.08 (m, 1H), 2.08 - 1.76 (m, 3H); MS (ES+): 459.4, (M+l); (ES-): 457.4, (M-l).

Scheme 76

Préparation of (S)-N-(3-chloro-2-fluorobenzyl)-l-(2-(4-(methylamino)-lH-pyrazolo[3,4-. d]pyrimidin-l -yl)acetyl)pyrrolidine-2-carboxamide (76b)

Compound 76b was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-(methylamino)-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetic acid (75c) (0.08 g, 0.386 mmol) in DMF (3 mL) using TFA sait of (S)-N-(3-chloro-2-fluorobenzyl)pyrrolidine-2-carboxamide (76a) (0.143 g, 0.386 mmol; prepared according to the procedure reported by Kotian, Pravin L. et al., PCT Int. Appl. (2019), WO 2019195720 Al 20191010.), HATU (0.161 g, 0.425 mmol), DIPEA (0.202 mL, 1.158 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with CMA-80 in chloroform from 0 - 40%] (S)-N-(3-chloro-2-fluorobenzyl)-l-(2-(4-(methylamino)-lHpyrazolo[3,4-d]pyrimidin-l-yl)acetyl)pyrrolidine-2-carboxamide (76b) (59 mg, 34% yield) as a white solid; 1H NMR (300 MHz, DMSO-t/₆) δ 8.46 (t, J = 5.9 Hz, 1H), 8.32 - 8.19 (m, 2H), 8.06 (s, 1H), 7.45 (t, J = 7.5 Hz, 1H), 7.20 (t, J = 7.7 Hz, 1H), 7.08 (t, J = 7.9 Hz, 1H), 5.22 (s, 2H), 4.46 - 4.20 (m, 3H), 3.80 - 3.57 (m, 2H), 2.99 (d, J = 4.6 Hz, 3H), 2.17 -1.70 (m, 4H); ¹⁹F NMR (282 MHz, DMSO-<7₆) δ -121.60; MS (ES+): 446.5 (M+l).

Scheme 77

Préparation of (S)-l-(2-(4-amino-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetyl)-N-(6bromopyridin-2-yl)pyrrolidine-2-carboxamide (77b)

Compound 77b was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetic acid (77a) (0.05 g, 0.259 mmol; prepared according to the procedure reported by Kotian, Pravin L. et al., PCT Int. Appl. (2017), WO 2017136395 Al 20170810; incorporated by reference) in DMF (3 mL) using (S)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (13a) (0.070 g, 0.259 mmol), HATU (0.108 g, 0.285 mmol), DIPEA (0.054 mL, 0.311 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with CMA-80 in chloroform from 0 - 40%] (S)-l-(2-(4-amino-lH-pyrazolo[3,4-d]pyrimidinl-yl)acetyl)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (77b) (43 mg, 37% yield) as a white solid; Ή NMR (300 MHz, DMSO-<7₆) δ 10.89 (s, 1H), 8.14 (s, 1H), 8.06 (s, 1H), 8.03 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 7.9 Hz, 1H), 7.32 (d, J= 7.6 Hz, 1H), 5.23 (s, 2H), 4.56 15 4.45 (m, 1H), 3.79 - 3.66 (m, 2H), 2.24 - 2.08 (m, 1H), 2.06 - 1.80 (m, 3H); MS (ES+):

445.4 (M+l).

Scheme 78

Préparation of (S)-1 -(2-(4-amino-1 H-pyrazoIo[3,4-d]pyrimidin-1 -yl)acetyl)-N-(3 -chloro-220 fluorobenzyl)pyrrolidine-2-carboxamide (78a)

Compound 78a was prepared according to the procedure reported in step-3 of scheme-1, from

2-(4-amino-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetic acid (77a) (0.05 g, 0.259 mmol) in

197

DMF (3 mL) using (S)-N-(3-chloro-2-fluorobenzyI)pyrrolidine-2-carboxamide (76a) (0.096 g, 0.259 mmol), HATU (0.108 g, 0.285 mmol) and DIPEA (0.136 mL, 0.777 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), CMA-80 in chloroform from 0 - 40%] (S)-l-(2-(4-amino5 1 H-pyrazolo[3,4-d]pyrimidin-1 -yl)acetyl)-N-(3 -chloro-2-fluorobenzyl)pyrrolidine-2carboxamide (78a) (43 mg, 37% yield) as a white solid; The free based was converted to HCl sait by dissolving in acetonitrile and 0.1% aq. HCl followed by lyophilization to afford (S)-l(2-(4-amino-1 H-pyrazolo [3,4-d]pyrimidin-1 -yl)acetyl)-N-(3 -chloro-2fluorobenzyl)pyrroIidine-2-carboxamide (78a) HCl sait as a white solid; 'H NMR (300 MHz, 10 DMSO-i/d) δ 8.46 (t, J = 5.9 Hz, 1H), 8.13 (s, 1H), 8.09 (s, 1H), 7.45 (t, J = 7.6 Hz, 1H), 7.20 (t, J = 8.0 Hz, 1H), 7.08 (t, J = 7.9 Hz, 1H), 5.22 (s, 2H), 4.47 - 4.21 (m, 3H), 3.80 - 3.55 (m, 2H), 2.17 -1.72 (m, 4H); 19F NMR (282 MHz, DMSO-ίήί) δ -121.60; MS (ES+): 432.5 (M+l), MS (ES-): 466.4 (M+Cl).

Scheme 79

Préparation of (lR,3S,5R)-2-(2-(4-amino-lH-pyrazolo[3,4-d]pyrimidin-l-yI)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3 -carboxamide (79a)

Compound 79a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetic acid (77a) (279 mg, 0.909 mmol) in

DMF (15 mL) using (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (4a) (300 mg, 0.757 mmol), HATU (432 mg, 1.136 mmol), DIPEA (0.660 mL, 3.79 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0 - 50%] (lR,3S,5R)-2-(2-(4-amino-lH-pyrazolo[3,4-d]pyrimidin-l25 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (79a) (202 mg, 58% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/₆) δ 10.79 (s, 1H), 10.00 (s, 1H), 9.00 (s, 1H), 8.49 (s, 1H), 8.47 (s, 1H), 8.02 (d, J= 8.1 Hz, 1H), 7.72 (t, J= 8.0

198

Hz, 1H), 7.33 (dd, J= Ί.Ί, 0.7 Hz, 1H), 5.78 - 5.27 (m, 2H), 4.41 (dd, J= 9.0, 5.5 Hz, 1H), 3.86-3.76 (m, 1H), 2.38-2.11 (m, 2H), 1.95- 1.80 (m, 1H), 1.05-0.92 (m, 1H), 0.700.60 (m, 1H); MS (ES+): 457.00 (M+l); (ES-): 455.00 (M-l).

Scheme 80

Préparation of (lR,3S,5R)-2-(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (80a)

Compound 80a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (5c) (0.06 g, 0.312 mmol) in DMF (2 mL) using (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (4a) (0.118 g, 0.312 mmol), HATU (0.142 g, 0.375 mmol), DIPEA (0.164 mL, 0.937 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH:EtOAc (9:1) in hexanes 0 to 100%] (lR,3S,5R)-2-(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyI)-N-(615 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (80a) (50 mg, 35% yield) as a white solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 10.78 (s, 1H), 8.07 - 8.00 (m, 2H), 7.72 (t, J = 8.0 Hz, 1H), 7.33 (d, J= 7.7 Hz, 1H), 7.08 (d, J= 3.5 Hz, 1H), 6.99 (brs, 2H), 6.52 (d, J= 3.5 Hz, 1H), 5.38-5.05 (m, 2H), 4.47 - 4.38 (m, 1H), 3.77 - 3.68 (m, 1H), 2.34 - 2.11 (m, 2H), 1.94-1.79 (m, 1H), 1.06-0.95 (m, 1H), 0.69 - 0.61 (m, 1H); MS (ES+) 456.2 (M+l), MS (ES-) 454.3, (M-l); Analysis calculated for Ci9Hi8BrN₇O2.H₂O: C, 48.11; H, 4.25; N, 20.67;

Found: C, 47.86; H, 4.22; N, 20.27.

Scheme 81

199

Préparation of ( 1 R,3S,5R)-2-(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyI)-N-(3chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (81a)

Compound 81a was prepared according to the procedure reported in step-3 of scheme-1, from 5 2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (5c) (0.075 g, 0.392 mmol) in DMF (3 mL) using TFA sait of (lR,3S,5R)-N-(3-chloro-2-fluorobenzyl)-2azabicyclo[3.1.0]hexane-3-carboxamide (66a) (0.15 g, 0.392 mmol), HATU (0.179 g, 0.470 mmol), DIPEA (0.205 mL, 1.176 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in

DCM from 0 to 50%) (lR,3S,5R)-2-(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N(3-chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (Sla) (65 mg, 37% yield) as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.47 (t, J= 5.9 Hz, 1H), 8.08 (s, 1H), 7.50 - 7.41 (m, 1H), 7.29 (s, 2H), 7.25 - 7.17 (m, 1H), 7.16 - 7.07 (m, 2H), 6.61 (d, J= 3.5 Hz, 1H), 5.30 (d, J= 17.1 Hz, 1H), 5.15 (d, J= 17.1 Hz, 1H), 4.33 (dd, J= 11.4, 6.0 Hz,

1H), 4.25 (q, J=4.5 Hz, 1H), 3.69-3.58 (m, 1H), 2.30-2.18 (m, 1H), 2.18-2.06 (m, 1H),

1.90 - 1.76 (m, 1H), 1.06 - 0.96 (m, 1H), 0.73 - 0.63 (m, 1H); ¹⁹F NMR (282 MHz, DMSOd₆) δ -121.78; MS (ES+) 443.2 (M+l); MS (ES-): 477.2 (M+CI); Analysis Calculated for C21H20CIFN6O2.I.25H2O: C, 54.20; H, 4.87; N, 18.06; Found: C, 54.13; H, 4.90; N, 17.69.

Scheme 82

Préparation of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-(2-(4-(methylamino)-lH-pyrazolo[3,4d]pyrimidin-l-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (82a)

200

Compound 82a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-(methylamino)-lH-pyrazoIo[3,4-d]pyrimidin-l-yl)acetic acid (75c) (0.06 g, 0.290 mmol) in DMF (2 mL) using (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (0.110 g, 0.290 mmol), HATU (0.143 g, 0.376 mmol), DIPEA (0.152 mL, 0.869 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 to 80%] (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-(2-(4-(methylamino)-lHpyrazolo [3,4-d]pyrimidin-1 -yl)acetyl)-2-azabicyclo [3.1.0]hexane-3 -carboxamide (82a) (75 mg, 55% yield) as a white solid; Ή NMR (300 MHz, DMSO-î7₆) δ 10.78 (s, 1H), 8.63 (brs,

1H), 8.29 (s, 1H), 8.13 (s, 1H), 8.03 (d, J= 8.2 Hz, 1H), 7.72 (t, J= 8.0 Hz, 1H), 7.33 (d, J=

7.7 Hz, 1H), 5.64-5.16 (m, 2H), 4.40 (dd, J= 9.1, 5.5 Hz, 1H), 3.83-3.75 (m, 1H), 3.00 (d, J = 4.5 Hz, 3H), 2.35 - 2.11 (m, 2H), 1.94 - 1.78 (m, 1H), 1.02 - 0.92 (m, 1H), 0.67 - 0.55 (m, 1H); MS (ES+): 471.3, 473.3 (M+l), 493.2, 495.2 (M+Na).

Scheme 83

Préparation of (lR,3S,5R)-2-(2-(4-amino-lH-pyrrolo[2,3-b]pyridin-l-yl)acetyl)-N-(6bromopyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (83d)

Step-1: Préparation of ZerZ-butyl 2-(4-((to+-butoxycarbonyl)amino)-lH-pyrrolo[2,3b]pyridin-l-yl)acetate (83b)

201

Compound 83b was prepared according to the procedure reported in step-3 of scheme-17, from tert-buty! 2-(4-chloro-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (83a) (1.5 g, 5.62 mmol; CAS # 2243596-23-6; prepared according to the procedure reported in Kotian, P. L. et al., PCT Int. AppL (2017), WO 2017136395 Al 20170810) in toluene (40 mL) using XPhos (0.268 g, 0.562 mmol), t-butyl carbamate (0.988 g, 8.44 mmol), Pd₂(dba)3 (0.257 g, 0.281 mmol), césium carbonate (1.832 g, 5.62 mmol) and heating at 90 °C for 20 h. This gave after work up and purification teri-butyl 2-(4-((/er/-butoxycarbonyl)amino)-lH-pyrrolo[2,3b]pyridin-l-yl)acetate (83b) (1.4 g, 72% yield); ’H NMR (300 MHz, DMSO-î/ô) δ 9.73 (s, 1H), 8.05 (d, J= 5.5 Hz, 1H), 7.61 (d, J= 5.5 Hz, 1H), 7.32 (d, J= 3.6 Hz, 1H), 6.90 (d, J =

3.6 Hz, 1H), 4.94 (s, 2H), 1.52 (s, 9H), 1.40 (s, 9H); MS (ES+): 348.3 (M+l); (ES-): 346.3 (M-l).

Step-2: Préparation of 2-(4-amino-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (83c)

Compound 83c was prepared according to the procedure reported in step-2 of scheme-1, from tert-butÿl 2-(4-((ter/-butôxycarbonyl)ammo)-lH-pyrrolo[2,3-b]pyridm-l-yl)acetate (83b) (0.65 g, 2.159 mmol) using TFA (0.166 mL, 2.159 mmol) in DCM (10 mL) and stirring at

RT for 16 h. This gave after work up 2-(4-amino-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (83c) (1.05 g) TFA sait as a light-orange solid; MS (ES+): 192.1 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-lH-pyrrolo[2,3-b]pyridin-l-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (83d)

Compound 83d was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (83c) (0.09 g, 0.314 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (0.1 g, 0.314 mmol), HATU (0.143 g, 0.377 mmol), DIPEA (0.274 mL, 1.569 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (4 g), eluting with DMA-80 in DCM from 0 to 80%] (lR,3S,5R)-2-(2-(4-amino-lH-pyrrolo[2,3-b]pyridin-l-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (83d) (25 mg, 17% yield) as a white solid; Ή NMR (300 MHz, DMSO-t/₆) δ 10.76 (s, 1H), 8.04 (d, J= 8.2 Hz, 1H), 7.76 7.67 (m, 2H), 7.32 (d, J= 7.7 Hz, 1H), 7.05 (d, J= 3.5 Hz, 1H), 6.49 (d, J= 3.5 Hz, 1H), 6.24

- 6.11 (m, 3H), 5.30 (d, J= Π.0 Hz, 1H), 5.11 (d, J= 16.9 Hz, 1H), 4.42 (dd, J= 9.0, 5.5 Hz,

1H), 3.76-3.68 (m, 1H), 2.33-2.10 (m, 2H), 1.91-1.78 (m, 1H), 1.04-0.94 (m, 1H), 0.68 - 0.60 (m, 1H); MS (ES+): 455.2, 457.2 (M+l), 477.2, 479.2 (M+Na), MS (ES-): 453.2,

202

455.3 (M-l); Analysis calculated for C₂oHi9BrN₆0₂.H20; C, 50.75; H, 4.47; N, 17.76; Found C, 50.70; H, 4.53; N, 17.46.

Scheme 84

67a 84a

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (84a)

Compound 84a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (0.05 g, 0.259 mmol) in DMF (1 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (0.082 g, 0.259 mmol), HATU (0.118 g, 0.311 mmol), DIPEA (0.181 mL, 1.035 mmol) and stirring at RT for 2 days. This gave after workup and purification by flash column chromatography [silica gel (4 g), eluting with DMA-80 in DCM from 0 to 80%] (1R,3S,5R)2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (84a) (31 mg, 26% yield) as a white solid; ’H NMR (300 MHz, DMSO-ûfc) δ

10.82 (s, 1H), 8.12 (s, 1H), 8.07 (s, 1H), 8.03 (d, J= 8.2 Hz, 1H), 7.72 (t, J= 8.0 Hz, 1H),

7.33 (d, J= 7.7 Hz, 1H), 7.24 (s, 2H), 5.42 (d, J= 17.2 Hz, 1H), 5.15 (d, J= 17.1 Hz, 1H), 4.45 (dd, J =9.0, 5.4 Hz, 1H), 3.80 - 3.71 (m, 1H), 2.36-2.12 (m, 2H), 1.95-1.82 (m, 1H), 1.09 - 0.98 (m, 1H), 0.72 - 0.65 (m, 1H); MS (ES+): 457.2 (M+l), 479.2, 481.2 (M+Na).

Scheme 85

85a

85b

203

Préparation of ( 1 R,3S,5R)-2-(2-(4-amino-1 H-indol-1 -yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (85b)

Compound 85b was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-lH-indol-l-yl)acetic acid (85a) (0.08 g, 0.279 mmol) in DMF (2 mL) using

HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (0.089 g, 0.279 mmol), HATU (0.127 g, 0.334 mmol), DIPEA (0.243 mL, 1.393 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (4 g), eluting with DMA-80 in DCM from 0 to 40%] to afford ( 1 R,3 S,5R)-2-(2-(4-amino-1 H-indol-1 -yl)acetyl)-N-(6-bromopyridin-2-yl)-210 azabicyclo[3.1.0]hexane-3-carboxamide (85b) (18 mg, 14% yield) as a white solid; ^rH NMR (300 MHz, DMSO-î/ô) δ 10.77 (s, 1H), 8.03 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.35 - 7.29 (m, 1H), 7.05 (d, J= 3.2 Hz, 1H), 6.79 (t, J= 7.8 Hz, 1H), 6.56 - 6.46 (m, 2H), 6.15 (d, J= 7.4 Hz, 1H), 5.24 (d, J= 17.1 Hz, 1H), 5.18 (s, 2H), 5.03 (d, J= 17.1 Hz, 1H), 4.42 (dd, 9.0, 5.5 Hz, 1H), 3.80-3.71 (m, 1H), 2.33-2.10 (m, 2H), 1.92- 1.76 (m, 1H), 1.01

- 0.92 (m, 1H), 0.69 - 0.58 (m, 1H); MS (ES+): 454.2, 456.2 (M+l), MS (ES-): 452.2, 454.2 (M-l).

Scheme 86

5c 86b

Préparation of (lR,3S,5R)-2-(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-((S)-l20 (3-chloro-2-fluorophenyl)ethyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (86b)

Compound 86b was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (5c) (0.05 g, 0.260 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-((S)-l-(3-chloro-2-fluorophenyl)ethyl)-2azabicyclo[3.1.0]hexane-3-carboxamide (86a) (0.083 g, 0.260 mmol; prepared according to the procedure reported by Altmann, E. et al., PCT Int. Appl. (2012), WO 2012093101 Al 20120712; incorporated by reference), HATU (0.119 g, 0.312 mmol), DIPEA (0.182 mL,

204

1.041 mmol) and stirring at RT for 2 days. This gave after workup and purification by flash column chromatography [silica gel (4 g), eluting with DMA-80 in DCM from 0 to 80%] (lR,3S,5R)-2-(2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-((S)-l-(3-chloro-2fluorophenyl)ethyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (86b) (0.033 g, 28% yield) as a white solid; Ή NMR (300 MHz, DMSO-cfe) δ 8.35 (d, J= 7.7 Hz, 1H), 8.05 (s, 1H), 7.48 7.39 (m, 1H), 7.35 - 7.27 (m, 1H), 7.20 - 7.04 (m, 5H), 6.56 (d, J= 3.6 Hz, 1H), 5.35 - 4.96 (m, 3H), 4.26 (dd, J= 8.8, 4.9 Hz, 1H), 3.67 - 3.57 (m, 1H), 2.28 - 2.03 (m, 2H), 1.88 - 1.74 (m, 1H), 1.35 - 1.26 (m, 3H), 1.05 - 0.94 (m, 1H), 0.70 - 0.60 (m, 1H); ^{15 * * * 19 20}F NMR (282 MHz, DMSO-dô) δ -122.65; MS (ES+): 479.3, 481.2 (M+Na); MS (ES-): 491.3, 493.3 (M+Cl);

Analysis calculated for C22H22CIFN6O2.I.5H2O; C, 54.60; H, 5.21; N, 17.37; Found C, 54.97; H, 5.19; N, 17.39.

Scheme 87

Préparation of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-(2-(2,4-diamino-7H-pyrrolo[2,315 d]pyrimidin-7-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (87f)

Step-1: Préparation of tert-butyl 2-(2-amino-4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetate (87b)

Compound 87b was prepared according to the procedure reported in step-1 of scheme-1, from 4-chloro-7H-pyrrolo[2,3-d]pyrimidin-2-amine (87a) (6 g, 35.6 mmol; CAS# 65103520 58-4) in acetonitrile (400 mL) using ter/-butyl 2-bromoacetate (5.78 mL, 39.1 mmol), K2CO3 (9.84 g, 71.2 mmol) and heating at 65 °C for 48 h. This gave after workup and purification by

205 flash column chromatography [silica gel (120 g), eluting with EtOAc/MeOH (9:1) in Hexane from 0-100%] ZerZ-butyl 2-(2-amino-4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (87b) (1.51 g, 15% yield) as an off-white solid; ‘HNMR(300 MHz, DMSO-ίΖό) δ 7.14 (d, J = 3.7 Hz, 1H), 6.71 (s, 2H), 6.31 (d, J = 3.7 Hz, 1H), 4.78 (s, 2H), 1.41 (s, 9H).

Step-2: Préparation of ZerZ-butyl 2-(2-amino-4-azido-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (87c)

To a solution of ZerZ-butyl 2-(2-amino-4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (87b) (1.5 g, 5.31 mmol) in DMF (20 mL) was added sodium azide (1.725 g, 26.5 mmol) and heated at 80 °C for 10 h. The reaction was cooled to 25 °C, extracted with chloroform (3

X 50 mL) and the combined organics were dried, filtered and evaporated to dryness. The residue obtained was purified by flash column chromatography [silica gel (40 g), eluting with methanol in DCM from 0-100%] to afford ZerZ-butyl 2-(2-amino-4-azido-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetate (87c) (0.556 g, 36% yield) as an off-white solid; 'H NMR (300 MHz, DMSO-d_ô) δ 8.18 (s, 2H), 7.20 (d, J = 3.5 Hz, 1H), 6.76 (d, J = 3.5 Hz, 1H), 4.93 (s,

2H), 1.42 (s, 9H); MS (ES+): 290.3 (M+l); MS (ES-): 288.3 (M-l).

Step-3: Préparation of ZerZ-butyl 2-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (87d)

To a solution of ZerZ-butyl 2-(2-amino-4-azido-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (87c) (0.528 g, 1.825 mmol) in methanol (20 mL) was added palladium hydroxide on carbon (0.064 g, 0.091 mmol) and heated with stirring at 22 °C for 10 h. The reaction was cooled to °C and extracted with chloroform (3 x 50 mL). The combined organics were dried, filtered, and evaporated to dryness. The residue obtained was purified by flash column chromatography [silica gel (24 g), eluting with methanol in DCM from 0-100%] to fumish ZerZ-butyl 2-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (87d) (0.362 g, 75 % yield) as an off-white solid; MS (ES+): 264.3 (M+l); MS (ES-): 262.3 (M-l).

Step-4: Préparation of 2-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (87e)

Compound 87e was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (87d) (0.351 g, 1.333 mmol) using TFA (1.027 mL, 13.33 mmol) in DCM (10 mL) and stirring at RT for 12 h. This gave after workup 2-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (87e) (0.428 g, 100% yield) TFA sait as a yellow solid; MS (ES+): 208.2 (M+l).

206

Step-5 Préparation of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-(2-(2,4-diamino-7Hpyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (87f)

Compound 87f was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide trifluoro acetate (4a) (0.259 g, 0.654 mmol) in DMF (10 mL) using TFA sait of 2-(2,4-diamino-7Hpyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (87e) (0.21 g, 0.654 mmol), HATU (0.373 g, 0.981 mmol), DIPEA (0.911 mL, 5.23 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel 25 g] (lR,3S,5R)-N-(6bromopyridin-2-y 1)-2-(2-(2,4-diamino-7H-pyrrolo [2,3 -d]pyrimidin-7 -yl)acetyl)-210 azabicyclo[3.1.0]hexane-3-carboxamide (87f) (0.023 g, 8 % yield) as a white solid; *H NMR (300 MHz, DMSO-7₆) δ 11.92 (s, 1H), 10.83 (s, 1H), 8.37 (s, 1H), 8.03 (d, J= 8.2 Hz, 1H), 7.73 (t, J= 8.0 Hz, 1H), 7.34 (d, J= 7.7 Hz, 1H), 7.23 (s, 2H), 6.96 (d, J= 3.7 Hz, 1H), 6.60 (d, J= 3.7 Hz, 1H), 5.23 (d, J= 17.0 Hz, 1H), 4.96 (d, J= 17.0 Hz, 1H), 4.42 (dd, J= 9.0, 5.5 Hz, 1H), 3.75-3.61 (m, 1H), 2.36-2.11 (m, 2H), 1.94- 1.79 (m, 1H), 1.08-0.94 (m, 1H),

0.77 - 0.60 (m, 1H).

Scheme 88

Préparation of ( 1 R,3S,5R)-2-(2-(4-amino-1 H-pyrrolo[3,2-c]pyridin-1 -yl)acetyl)-N-(3-chloro2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (88f)

Step-1 : Préparation of tert-butyl 2-(4-chloro-lH-pyrrolo[3,2-c]pyridin-l-yl)acetate (88b)

207

To a solution of 4-chloro-lH-pyrrolo[3,2-c]pyridine (88a) (0.3 g, 1.966 mmol; CAS # 6029021-3) in DMF (10 mL) was added sodium hydride (0.079 g, 1.966 mmol) in portions at 0 °C under an argon atmosphère. After stirring for 15 min was added terributyl bromoacetate (0.436 mL, 2.95 mmol) drop wise and stirred at RT for 2h. Reaction mixture was poured into 5 sat. NH4CI solution and extracted with EtOAc (2 x 60 mL). The combined organics were washed with brine, dried, filtered and concentrated in vacuum. The residue obtained was purifïed by flash column chromatography [silica gel (40 g), eluting with EtOAc in hexane as eluents from 0 to 50%] to afford ZerZ-butyl 2-(4-chloro-lH-pyrrolo[3,2-c]pyridin-l-yl)acetate (88b) (0.45 g, 86 % yield) as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.01 (d, J = 5.8

Hz, 1H), 7.55 (d, J = 3.3 Hz, 1H), 7.52 (dd, J = 5.7, 0.9 Hz, 1H), 6.59 (dd, J = 3.3, 0.9 Hz, 1H), 5.13 (s, 2H), 1.41 (s, 9H).

Step-2: Préparation of 2-(4-chloro-lH-pyrrolo[3,2-c]pyridin-l-yl)acetic acid (88c)

Compound 88c was prepared according to the procedure reported in step-2 of scheme-1, from terributyl 2-(4-chloro-lH-pyrrolo[3,2-c]pyridin-l-yl)acetate (88b) (0.45 g, 1.687 mmol) using TFA (1.30 mL, 16.87 mmol) in DCM (10 mL) and stirring at RT for 16 h. This gave after workup 2-(4-chloro-lH-pyrrolo[3,2-c]pyridin-l-yl)acetic acid (88c) (0.34 g, 66% yield) TFA sait as an off-white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.01 (d, J = 5.8 Hz, 1H), 7.60 - 7.52 (m, 2H), 6.59 (dd, J = 3.2, 0.9 Hz, 1H), 5.13 (s, 2H).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-chloro-lH-pyrrolo[3,2-c]pyridin-l-yl)acetyl)-N20 (3-chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (88d)

Compound 88d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-chloro-lH-pyrrolo[3,2-c]pyridin-l-yl)acetic acid (88c) (0.13 g, 0.423 mmol) in DMF (3 mL) using HCl sait of (lR,3S,5R)-N-(3-chloro-2-fluorobenzyl)-2azabicyclo[3.1.0]hexane-3-carboxamide (66a) (0.129 g, 0.423 mmol), HATU (0.193 g, 0.507 25 mmol), DIPEA (0.328 g, 2.54 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (4 g), eluting with MeOH:EtOAc (9:1) in hexanes from 0 - 80%] (lR,3S,5R)-2-(2-(4-chloro-lH-pyrrolo[3,2-c]pyridin-lyl)acetyl)-N-(3-chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (88d) (0.16 g, 82% yield) as a white solid; ’H NMR (300 MHz, DMSO-îZ₆) δ 8.48 (t, J = 6.0 Hz, 30 1H), 7.95 (dd, J = 5.7, 1.2 Hz, 1H), 7.53 (d, J = 3.3 Hz, 1H), 7.51 - 7.41 (m, 2H), 7.18 (t, J =

7.3 Hz, 1H), 7.05 (t, J = 7.8 Hz, 1H), 6.58 (dd, J = 3.2, 1.0 Hz, 1H), 5.47 (d, J = 17.4 Hz, 1H), 5.27 (d, J = 17.3 Hz, 1H), 4.46 - 4.35 (m, 1H), 4.33 - 4.20 (m, 2H), 3.71 - 3.63 (m, 1H), 2.32

208

2.08 (m, 2H), 1.94 - 1.79 (m, 1H), 1.07 - 0.97 (m, 1H), 0.82 - 0.71 (m, 1H); MS (ES+): 461.2 (M+l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-lH-pyrrolo[3,2-c]pyridin-l-yl)acetyl)-N-(3chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (88f)

Compound 88f was prepared according to the procedure reported in step-3 of scheme-17, from ( 1 R,3S,5R)-2-(2-(4-chloro-1 H-pyrrolo[3,2-c]pyridin-1 -yl)acetyl)-N-(3-chloro-2fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (88d) (0.15 g, 0.325 mmol) in dioxane (5 mL) using XPhos (0.016 g, 0.033 mmol), /er/-butyl carbamate (88e) (0.076 g, 0650 mmol), Pd₂(dba)3 (0.015 g, 0.016 mmol), césium carbonate (0.106 g, 0.325 mmol) and heating at 90 °C for 20 h. This gave after work up and purification [silica gel (12 g), eluting withDMA-80 in DCM from 0 - 100%] (lR,3S,5R)-2-(2-(4-amino-lH-pyrrolo[3,2-c]pyridinl-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (88f) (0.01 g, 7% yield) as a white solid; Ή NMR (300 MHz, DMSCMû δ 8.52 - 8.41 (m, 1H), 7.57 - 7.40 (m, 2H), 7.27 - 7.15 (m, 1H), 7.14 - 7.02 (m, 2H), 6.70 - 6.57 (m, 2H), 6.19 (s, 2H), 5.25 (d, J= 17.3 Hz, 1H), 5.08 (d, J= 17.4 Hz, 1H), 4.48 - 4.18 (m, 3H), 3.70 - 3.61 (m, 1H), 2.32-2.06 (m, 2H), 1.92- 1.77 (m, 1H), 1.06-0.94 (m, 1H), 0.76-0.64 (m, 1H); 19F NMR (282 MHz, DMSO-î/₆) δ -121.81; MS (ES+) 442.3, 444.3 (M+l), MS (ES-) 476.3, 478.3 (M+Cl).

Scheme 89

Préparation of (lR,3S,5R)-2-(2-(2-amino-9H-purin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (89b)

Compound 89b was prepared according to the procedure reported in step-3 of scheme-1, from 2-(2-amino-9H-purin-9-yl)acetic acid (89a) (0.04 g, 0.207 mmol; CAS# 933477-63-5) in DMF (1 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (0.066 g, 0.207 mmol), HATU (0.094 g, 0.248

209

mmol), DIPEA (0.145 mL, 0.828 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (4 g), eluting with DMA-80 in DCM from 0 -100%] (lR,3S,5R)-2-(2-(2-amino-9H-purin-9-yl)acetyl)-N-(6-bromopyridin-2yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (89b) (62 mg, 66% yield) as a white solid; 'H

NMR (300 MHz, DMSOX) δ 10.85 (s, 1H), 8.58 (s, 1H), 8.07 - 7.97 (m, 2H), 7.72 (t, J= 8.0 Hz, 1H), 7.35-7.31 (m, 1H), 6.61 (s, 2H), 5.32 (d, J= 17.3 Hz, 1H), 5.06 (d, J= 17.3 Hz, 1H), 4.44 (dd, J= 9.0, 5.4 Hz, 1H), 3.76 - 3.68 (m, 1H), 2.37 - 2.11 (m, 2H), 1.95-1.81 (m, 1H), 1.10 - 0.96 (m, 1H), 0.74 - 0.63 (m, 1H); MS (ES+): 457.2, 459.2 (M+l), 479.2, 481.2 (M+Na), MS (ES-): 455.3 (M-l); Analysis calculated for CisHiTBrNgCh.^O; C, 45.49; H,

4.03; N, 23.58; Found C, 44.88; H, 4.12; N, 22.82.

Scheme 90

90d

Préparation of (lR,3S,5R)-2-(2-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (90d)

Step-1 : Préparation of ZerZ-butyl 2-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetate (90b)

Compound 90b was prepared according to the procedure reported in step-1 of scheme-1, from 2-chloro-7H-pyrrolo[2,3-d]pyrimidin-4-amine (90a) (1 g, 5.93 mmol; CAS # 119271188-8) in acetonitrile (50 mL) using ZerZ-butyl 2-bromoacetate (0.963 mL, 6.52 mmol) and

210

K2CO3 (1.640 g, 11.86 mmol) and heating at 65 °C for 14 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with EtOAc in Hexane from 0-100%] ZerZ-butyl 2-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetate (90b) (1.102 g, 66% yield) as a white solid; *H NMR (300 MHz, DMSO-i/g) δ 7.52 (s, 2H), 7.12 (d, J = 3.5 Hz, 1H), 6.55 (d, J = 3.5 Hz, 1H), 4.83 (s, 2H), 1.41 (s, 9H); MS (ES+): 283.2 (M+l); (ES-): 281.3 (M-l).

Step-2: Préparation of 2-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (90c)

Compound 90c was prepared according to the procedure reported in step-2 of scheme-1, from 10 ZerZ-butyl 2-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (90b) (0.251 g,

0.888 mmol) using TFA (0.684 mL, 8.88 mmol) in DCM (10 mL) and stirring at RT for 12 h. This gave after workup 2-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (90c) (0.302 g, 100% yield) TFA sait as a yellow solid; MS (ES+): 227.1 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-715 yl)acetyI)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (90d)

Compound 90d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (90c) (0.175 g, 0.514 mmol) in DMF (10 mL) using TFA sait of (lR,3S,5R)-N-(6-bromopyridin-2yl)-2-azabicycIo[3.1.0]hexane-3-carboxamide (4a) (0.204 g, 0.514 mmol), HATU (293 mg, 20 0.771 mmol), DIPEA (0.716 mL, 4.11 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (25 g), eluting with EtOAc in MeOH (9:1) from 0 - 100%] (lR,3S,5R)-2-(2-(4-amino-2-chloro-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetyI)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (90d) (132 mg, 52% yield) as a white solid; 'H NMR (300 MHz, DMSOY₆) δ 10.81 (s, 1H,

D₂O exchangeable), 8.04 (d, J= 8.2 Hz, 1H), 7.72 (t, J= 8.0 Hz, 1H), 7.48 (s, 2H, D₂O exchangeable), 7.33 (dd, J= 7.8, 0.7 Hz, 1H), 7.08 (d, J= 3.5 Hz, 1H), 6.53 (d, J= 3.5 Hz, 1H), 5.31 (d, J= 17.1 Hz, 1H), 5.03 (d, J= 17.1 Hz, 1H), 4.43 (dd, J= 9.0, 5.5 Hz, 1H), 3.82 -3.68(m, 1H), 2.36-2.11 (m, 2H), 1.93-1.75 (m, 1H), 1.07-0.94 (m, 1H), 0.74-0.62 (m, 1H); Analysis calculated for Ci9Hi7BrClN7O2.H₂O: C, 44.86; H, 3.76; N, 19.27; Found:

C, 44.79; H, 3.67; N, 19.12.

Scheme 91

211

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)2-azabicyclo [3.1.0]hexane-3 -carboxamide (91a)

Compound 91a was prepared according to the procedure reported in step-3 of scheme-1, from 5 TFA sait of 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (0.188 g, 0.612 mmol) in DMF (15 mL) using HCl sait of (lR,3S,5R)-N-(3-chIoro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3carboxamide (66a) (0.224 g, 0.734 mmol), HATU (349 mg, 0.918 mmol), DIPEA (0.533 mL, 3.06 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 15%] (1 R,3 S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3 -chloro-2-fluorobenzyl)-2azabicyclo[3.1.0]hexane-3-carboxamide (91a) (101 mg, 37% yield) as a white solid; 'H NMR (300 MHz, DMSO-îZô) δ 8.47 (t, J= 6.0 Hz, 1H), 8.11 (s, 1H), 8.04 (s, 1H), 7.51 - 7.40 (m, 1H), 7.27 - 7.16 (m, 3H), 7.10 (t, J= 7.9 Hz, 1H), 5.44 - 5.10 (m, 2H), 4.48 - 4.19 (m, 3H), 3.77-3.59 (m, 1H), 2.34-2.05 (m, 2H), 1.94-1.76 (m, 1H), 1.13-0.96 (m, 1H), 0.77 15 0.65 (m, 1H); ¹⁹F NMR (282 MHz, DMSO<fc) δ -121.75; MS (ES⁺): 444.10 (M + 1); MS (ES’): 442.10 (M - 1).

Scheme 92

92b

212

Préparation of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-(2-(2,6-diamino-9H-purin-9yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (92b)

Compound 92b was prepared according to the procedure reported in step-3 of scheme-1, from 2-(2,6-diamino-9H-purin-9-yl)acetic acid (92a) (0.1 g, 0.328 mmol) in DMF (1 mL) and using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (4a) (0.104 g, 0.328 mmol), HATU (0.150 g, 0.393 mmol), DIPEA (0.343 mL, 1.966 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (4 g), eluting with DMA-80 in DCM from 0 - 100%] (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-(2-(2,6-diamino-9H-purin-9-yl)acetyl)-210 azabicyclo[3.1.0]hexane-3-carboxamide (92b) (0.06 g, 39% yield) as a white solid; Ή NMR (300 MHz, DMSO-7ô) δ 10.84 (s, 1H), 8.04 (d, J= 8.2 Hz, 1H), 7.72 (t, J= 8.0 Hz, 1H), 7.63 (s, 1H), 7.33 (d, J= 7.7 Hz, 1H), 6.69 (brs, 2H), 5.81 (brs, 2H), 5.19 (d, J= 17.3 Hz, 1H), 4.94 (d, J= 17.3 Hz, 1H), 4.44 (dd, J= 9.1, 5.6 Hz, 1 H), 3.75 - 3.64 (m, 1H), 2.34-2.10 (m, 2H), 1.94 - 1.79 (m, 1H), 1.07 - 0.96 (m, 1H), 0.72 - 0.60 (m, 1H); MS (ES+): 494.2, 496.2 (M+Na); MS (ES-): 470.3, 472.3 (M-l).

Scheme 93 ^N\-^N^Br

HATU, DIPEA

Préparation of (2S,4R)-1 -(2-(6-amino-9H-purin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4fIuoropyrrolidine-2-carboxamide (93a)

Compound 93a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (0.466 mmol, 190 mg) in DMF (15 mL) and using TFA sait of (2S,4R)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2carboxamide (5d) (0.559 mmol, 250 mg), HATU (266 mg, 0.698 mmol), DIPEA (0.405 mL, 2.328 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 25%]

213 (2S,4R)-l-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine2-carboxamide (93a) (129 mg, 60% yield) as a white solid; *H NMR (300 MHz, DMSO-ô/ô) δ 11.04 (s, 1H), 8.11 (s, 1H), 8.07-7.99 (m, 2H), 7.71 (t, 7=8.0 Hz, 1H), 7.36-7.29 (m, 1H), 7.21 (s, 2H), 5.65 - 5.36 (m, 1H), 5.32 - 5.00 (m, 2H), 4.66 (t, J= 8.5 Hz, 1H), 4.29 - 3.74 5 (m, 2H), 2.67 - 1.95 (m, 2H); ¹⁹F NMR (282 MHz, DMSO-î7₆) δ -175.72; MS (ES+): 463.00 & 465.00 (M+l); MS (ES-): 461.00 & 463.00 (M-l); Analysis calculated for

Ci₇Hi₆BrFN₈O2.2.0 H₂O: C, 40.89; H, 4.04; N, 22.44; Found: C, 41.06; H, 4.05; N, 22.30.

Scheme 94

Préparation of (lR,3S,5R)-benzyl 2-(2-(6-amino-9H-purin-9-yl)acetyl)-2azabicyclo[3.1.0]hexane-3 -carboxylate (94b)

Compound 94b was prepared according to the procedure reported in step-3 of scheme-1, from 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (2.127 g, 11.01 mmol) in DMF (30 mL) and using TFA sait of (lR,3S,5R)-benzyl 2-azabicyclo[3.1.0]hexane-3-carboxylate (94a) (4.56 g, 13.76 mmol; CAS # 1386459-65-9), HATU (6.28 g, 16.52 mmol), DIPEA (7.67 mL, 44.0 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), DMA-80 in DCM from 0 - 50%] (lR,3S,5R)-benzyl 2-(2(6-amino-9H-purin-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxylate (94b) (2.83 g, 65.5 % yield) as a yellow solid; Ή NMR (300 MHz, DMSO-7₆) δ 8.13 (s, 1H), 8.06 (s, 1H), 7.41 20 7.24 (m, 7H), 5.43 (d, J = 17.2 Hz, 1H), 5.20 - 5.03 (m, 3H), 4.33 (dd, J = 9.3, 5.6 Hz, 1H),

3.77 (ddd, J = 7.1, 5.5, 2.4 Hz, 1H), 2.42 - 2.28 (m, 1H), 2.23 - 2.09 (m, 1H), 1.94 - 1.83 (m, 1H), 1.07 - 0.94 (m, 1H), 0.81 - 0.71 (m, 1H); MS (ES+): 393.1 (M+l); Analysis calculated for C20H20N6O3.2H2O.I.75HCI: C, 48.80; H, 5.27; N, 17.07; Found: C, 48.78; H, 5.17; N, 17.01.

Scheme 95

214

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3,3-dimethylbutyl)-2azabicyclo[3.1.0]hexane-3-carboxamide (95c)

Step-1: Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-25 azabicyclo[3.1.0]hexane-3-carboxylic acid (95a)

To a solution of (lR,3S,5R)-benzyl 2-(2-(6-amino-9H-purin-9-yl)acetyl)-2azabicyclo[3.1.0]hexane-3-carboxylate (94b) (300 mg, 0.765 mmol) in MeOH (20 mL) was added 10% Pd/C (100 mg, 0.093 mmol). The reaction mixture was shaken for 5 hours under a hydrogen atmosphère (50-55 psi), filtered through a pad of Celite and the pad was washed with methanol. The filtrate was concentrated in vacuum and residue obtained was purified by reverse-phase column chromatography [C-18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] to afford (lR,3S,5R)-2-(2-(6-amino-9H-purin-9yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (95a) (145 mg, 63% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-ώ) δ 9.62 (s, 1H), 8.88 (s, 1H), 8.52 (s, 1H),

8.46 (s, 1H), 5.57 (d, J = 17.2 Hz, 1H), 5.28 (d, J = 17.1 Hz, 1H), 4.20 (dd, J = 9.2, 5.2 Hz,

1H), 3.72 (td, J = 6.2, 5.2, 2.3 Hz, 1H), 2.38 - 2.25 (m, 1H), 2.19 (dq, J = 13.0, 6.4, 5.8 Hz, 1H), 1.28 (dd, J = 9.8, 6.5 Hz, 1H), 1.09-0.91 (m, 1H), 0.73 (td, J = 5.1, 2.5 Hz, 1H). MS (ES+): 303.1 (M+l), (ES-): 301.1 (M-l).

Step-2: Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3,320 dimethylbutyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (95c)

Compound 95c was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (95a) 135mg, 0.447 mmol) in DMF (1.5 mL) using HCl sait of 3,3-dimethylbutan-lamine (95b) (4.56 g, 13.76 mmol; CAS # 30654-98-8), HATU (255 mg, 0.670 mmol), DIPEA (0.311 mL, 1.786 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] followed by purification using flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 50%] (1R,3S,5R)2-(2-(6-amino-9H-purin-9-yI)acetyl)-N-(3,3-dimethylbutyl)-2-azabicyclo[3.1,0]hexane-330 carboxamide(95c) (25 mg, 15%yield) as awhitesolid; 'HNMRQOOMHz,DMSO-î4>)ô 8.12

215 (s, 1H), 8.05 (s, 1H), 7.75 (t, J = 5.7 Hz, 1H), 7.22 (s, 2H), 5.35 (d, J = 17.1 Hz, 1H), 5.13 (d, J = 17.0 Hz, 1H), 4.18 (dd, J= 8.3, 5.0 Hz, 1H), 3.65-3.59 (m, 1H), 3.10-2.96 (m, 2H), 2.24 -2.08 (m, 2H), 1.81 (s, 1H), 1.33 - 1.22 (m, 2H), 1.10 - 0.97 (m, 1H), 0.87 (s, 9H), 0.73 0.63 (m, 1H); MS (ES+): 386.2 (M+l); Analysis calculated for C19H27N7O2.I.25H2O: C, 55.93;

H, 7.29; N, 24.03; Found: C, 56.04; H, 7.35; N, 23.94.

Scheme 96

Préparation of (lR,3S,5R)-2-(2-(6-amino-2-(dimethylamino)-9H-purin-9-yl)acetyl)-N-(3chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (96e)

Step-1 : Préparation of N2,N2-dimethyl-7H-purine-2,6-diamine (96b)

To a stirred suspension of 2-chloro-7H-purin-6-amine (96a) (1 g, 5.90 mmol; CAS# 1839-185) in isopropanol (10 mL) was added triethylamine (1.644 mL, 11.79 mmol) and dimethylamine (0.717 mL, 7.08 mmol) and heated in a microwave atl50 °C for 1.5 h. The solid separated was collected by filtration to afford N2,N2-dimethyl-7H-purine-2,6-diamine (96b) (750mg, 71.4 % yield) as a white solid; Ή NMR (300 MHz, DMSO-ûfc) δ 12.19 (s,

1H), 7.67 (s, 1H), 6.65 (s, 2H), 3.05 (s, 6H); MS (ES+): 179.10 (M+l).

Step-2: Préparation of ZerZ-butyl 2-(6-amino-2-(dimethylamino)-9H-purin-9-yl)acetate (96c)

Compound 96c was prepared according to the procedure reported in step-1 of scheme-1, from N2,N2-dimethyl-7H-purine-2,6-diamine (96b) (500 mg, 2.81 mmol) inN,N20 dimethylformamide (5 mL) using ZerZ-butyl 2-bromoacetate (0.498 mL, 3.37 mmol), K2CO3 (582 mg, 4.21 mmol) and stirring at RT for 10 h. This gave after workup and purification by flash column chromatography [silica gel (25 g), eluting with EtOAc/MeOH (9:1) in Hexane

216 from 0-100%] tert-butyl 2-(6-amino-2-(dimethylamino)-9H-purin-9-yl)acetate (96c) (620 mg, 76% yield) as a bright yellow solid; *H NMR (300 MHz, DMSO-î/₆) δ 7.68 (s, 1H), 6.74 (s, 2H), 4.75 (s, 2H), 3.04 (s, 6H), 1.42 (s, 9H); MS (ES+): 293.20 (M+l).

Step-3: Préparation of 2-(6-amino-2-(dimethylamino)-9H-purin-9-yl)acetic acid (96d)

Compound 96d was prepared according to the procedure reported in step-4 of scheme-17, from tert-butyl 2-(6-amino-2-(dimethylamino)-9H-purin-9-yl)acetate (96c) (600 mg, 2.052 mmol) in THF (5 mL), methanol (5.0 mL), water (5 mL) using lithium hydroxide (147 mg, 6.16 mmol) and stirring ovemight at RT. This gave after workup 2-(6-amino-2(dimethylamino)-9H-purin-9-yl)acetic acid (96d) (352 mg, 73 % yield) as a yellow solid; ’H

NMR (300 MHz, DMSO-d₆) δ 7.70 (s, 1H), 6.75 (s, 2H), 4.77 (s, 2H), 3.05 (s, 6H).

Step-4: Préparation of (lR,3S,5R)-2-(2-(6-amino-2-(dimethylamino)-9H-purin-9-yl)acetyl)N-(3-chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (96e)

Compound 96e was prepared according to the procedure reported in step-3 of scheme-1, from 2-(6-amino-2-(dimethylamino)-9H-purin-9-yl)acetic acid (96d) (150 mg, 0.635 mmol) in

DMF (3 mL) using HCl sait of (lR,3S,5R)-N-(3-chloro-2-fluorobenzyl)-2azabicyclo[3.1.0]hexane-3-carboxamide (66a) (242 mg, 0.794 mmol), HATU (362 mg, 0.952 mmol), DIPEA (0.442 mL, 2.54 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with EtOAc/MeOH (9:1) in dioxane from 0 -100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] to afford (lR,3S,5R)-2-(2-(6-amino-2-(dimethylamino)-9H-purin-9yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (96e) (240 mg, 78% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-cfc + D₂O) δ 8.57 (t, J = 6.0 Hz, 1H), 8.16 (d, J= 6.1 Hz, 1H), 7.45 (t, J= 7.6 Hz, 1H), 7.21 (t, J= 7.3 Hz, 1H), 7.10 (t,J= 7.9 Hz, 1H), 5.29 (dd, J= 17.1, 3.1 Hz, 1H), 5.15 (d, J= 17.0 Hz, 1H), 4.33 (t, J= 6.9

Hz, 2H), 4.28 - 4.19 (m, 1H), 3.13 (s, 6H), 2.28 (dd, J= 13.5, 9.2 Hz, 1H), 2.14 (dt, J= 12.9, 6.0 Hz, 1H), 1.88 (p, J= 7.1 Hz, 2H), 1.07 (dt, J= 9.5, 5.4 Hz, 1H), 0.68 (d, J= 6.0 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-t/₆) δ -121.73; MS (ES+): 487.2 (M+l); (ES-): 485.10 (M-l).

Scheme 97

217

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3,3-dimethylcyclohexyl)2-azabicyclo[3.1.0]hexane-3-carboxamide (97b)

Compound 97b was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3carboxylic acid (95a) 150mg, 0.496 mmol) in DMF (1.5 mL) using 3,3dimethylcyclohexanamine (97a) (63.1 mg, 0.496 mmol; CAS # 226549-07-1), HATU (283 mg, 0.744 mmol) DIPEA (0.346 mL, 1.985 mmol) and stirring at RT for 2 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with

DMA-80 in DCM from 0-100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3,3-dimethylcyclohexyl)-2azabicyclo[3.1.0]hexane-3-carboxamide (97b) (20mg, 10% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d6) δ 8.49 (s, 1H), 8.43 (s, 1H), 7.58 (d, J = 8.0 Hz, 1H), 5.54 (d, J= 17.1 Hz, 1H), 5.24 (d, J= 17.1 Hz, 1H), 4.14 (dd, J= 8.9, 4.9 Hz, 1H), 3.70-3.65 (m,

2H), 2.28-2.00 (m, 2H), 1.95-1.77 (m, 1H), 1.77- 1.60 (m, 1H), 1.60-1.16 (m, 4H), 1.11 - 0.91 (m, 4H), 0.87 (s, 6H), 0.74 - 0.56 (m, 1H); MS (ES+): 412.2 (M+l).

Scheme 98

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-((lR,2S)-2-(2chlorophenyI)cyclopropyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (98b)

218

Compound 98b was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3carboxylic acid (95a) (150mg, 0.496 mmol) in DMF (1.5 mL) using (lR,2S)-2-(2chlorophenyl)cyclopropanamine (98a) (83 mg, 0.496 mmol; CAS # 1820575-68-5), HATU (283 mg, 0.744 mmol), DIPEA (0.346 mL, 1.985 mmol) and stirring at RT for 2 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCI) from 0-100%] (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-((lR,2S)-2-(210 chlorophenyl)cyclopropyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (98b) (39 mg, 17 % yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-î4>) δ 8.50 — 8.33 (m, 2H), 8.23 (d, J= 4.8 Hz, 1H), 7.40 (dt, J= 7 A, 1.8 Hz, 1H), 7.31 - 7.14 (m, 2H), 7.08 (dd, J= 7 A, 2.1 Hz, 1H), 5.54 (d, J= 17.1 Hz, 1H), 5.25 (d, J = 17.0 Hz, 1H), 4.22-4.11 (m, 1H), 3.75 3.64 (m, 1H), 3.01 - 2.88 (m, 1H), 2.29 - 2.07 (m, 3H), 1.89 - 1.83 (m, 1H), 1.30 - 1.17 (m,

1H), 1.17 - 1.04 (m, 1H), 1.08 - 0.97 (m, 1H), 0.72 - 0.65 (m, 1H); MS (ES+) 452.1 (M+l);

Analysis calculated for C22H22CIN7O2.HCI.3H2O: C, 48.71; H, 5.39; Cl, 13.07; N, 18.08; Found: C, 48.61; H, 5.22; Cl, 13.33; N, 18.00.

Scheme 99

F

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yI)acetyl)-N-(2(trifluoromethoxy)ethyl)-2-azabicyclo [3.1.0]hexane-3-carboxamide (99b)

Compound 99b was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (95a) (150 mg, 0.496 mmol) in DMF (1.5 mL) using 2-(trifhioromethoxy)ethanamine (99a) (64.1 mg, 0.496 mmol; CAS # 886050-51-7), HATU (283 mg, 0.744 mmol), DIPEA (0.346 mL, 1.985 mmol) and stirring at RT for 2 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 -100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g),

219 eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-amino9H-purin-9-yI)acetyl)-N-(2-(trifluoromethoxy)ethyl)-2-azabicyclo[3.1.0]hexane-3carboxamide (99b) (17 mg, 8% yield) as a white solid; 'H NMR (300 MHz, DMSO-î/ô) δ 9.55 (s, 1H), 8.86 (s, 1H), 8.51 (s, 1H), 8.45 (s, 1H), 8.26 (t, J= 5.7 Hz, 1H), 5.57 (d, J= 17.1 Hz, 5 1H), 5.25 (d, J= 17.0 Hz, 1H), 4.22 (dd, J= 9.0, 4.6 Hz, 1H), 4.04 (t, J= 5.4 Hz, 2H), 3.69 3.64 (m, 1H), 3.39 - 3.16 (m, 2H), 2.30 - 2.01 (m, 2H), 1.89 - 1.74 (m, 1H), 1.11 - 0.94 (m, 1H), 0.78 - 0.61 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-d6) δ -58.83; MS (ES+): 414.1 (M+l).

Scheme 100

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-((3methylcyclobutyl)methyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (100b)

Compound 100b was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3carboxylic acid (95a) (150 mg, 0.496 mmol) in DMF (1.5 mL) using (375 methylcyclobutyl)methanamine (100a) (98 mg, 0.992 mmol; CAS # 1445951-46-1), HATU (283 mg, 0.744 mmol), DIPEA (0.346 mL, 1.985 mmol) and stirring at RT for 3 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from

0-100%] (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yI)acetyl)-N-((3-methyIcyclobutyl)methyl)2-azabicyclo[3.1.0]hexane-3-carboxamide (100b) (19 mg, 10% yield) HCl sait as a white solid; ‘H NMR (300 MHz, DMSO-tZ₆) δ 9.61 (s, 1H), 8.87 (s, 1H), 8.52 (s, 1H), 8.45 (s, 1H), 7.95 - 7.76 (m, 1H), 5.57 (dd, J= 17.2, 1.3 Hz, 1H), 5.26 (dd, J= 17.0, 1.5 Hz, 1H), 4.30 4.14 (m, 1H), 3.78 - 3.60 (m, 1H), 3.26 - 2.86 (m, 2H), 2.44 - 1.90 (m, 3H), 1.93 - 1.70 (m,

2H), 1.62 - 1.42 (m, 2H), 1.39 - 0.82 (m, 6H), 0.78 - 0.61 (m, 1H); MS (ES+): 384.2 (M+l).

220

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-benzyl-2azabicycio [3.1.0]hexane-3 -carboxamide (101b)

Compound 101b was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3carboxylic acid (95a) (150 mg, 0.496 mmol) in DMF (1.5 mL) using phenylmethanamine (101a) (80 mg, 0.744 mmol; CAS # 100-46-9), HATU (283 mg, 0.744 mmol), DIPEA (0.346 mL, 1.985 mmol) and stirring at RT for 2 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-amino9H-purin-9-yl)acetyl)-N-benzyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (101b) (51 mg, 26% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-cfe) δ 9.38 (s, 1H), 8.73 (s,

1H), 8.52 - 8.44 (m, 2H), 8.43 (s, 1H), 7.36 - 7.17 (m, 5H), 5.55 (d, J= 17.1 Hz, 1H), 5.27 (d,

J= 17.1 Hz, 1H), 4.35 - 4.19 (m, 3H), 3.73 - 3.67 (m, 1H), 2.33 - 2.10 (m, 2H), 1.93 -1.84 (m, 1H), 1.13 - 1.01 (m, 1H), 0.77 - 0.67 (m, 1H); MS (ES+): 392.2 (M+l).

Scheme 102

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3,3-dichlorocyclohexyl)2-azabicyclo[3.1.0]hexane-3-carboxamide (102b)

Compound 102b was prepared according to the procedure reported in step-3 of scheme-1, from ( 1 R,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-321436

221 carboxylic acid (95a) (90 mg, 0.298 mmol) in DMF (1 mL) using 3,3dichlorocyclohexanamine (102a) (50 mg, 0.298 mmol; CAS # 226549-07-1), HATU (170 mg, 0.446 mmol), DIPEA (0.207 mL, 1.190 mmol) and stirring at RT for 2 h. This gave after workup and purification twice by reverse phase column chromatography [Cl8 column (50 g), 5 eluting with ACN in water (containing 0.1% HCl) from 0-100%] followed by purification twice using flash column chromatography [silica gel (12 g), DMA80 in DCM from 0 - 100%] (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3,3-dichlorocyclohexyl)-2azabicyclo[3.1.0]hexane-3-carboxamide (102b) (10 mg, 7% yield) as a white solid; 'H NMR (300 MHz, DMSO-î/ô) δ 8.15 (s, 1H), 8.08 (d, J= 3.6 Hz, 1H), 7.80 (dd, J= 8.1, 2.1 Hz, 1H), 10 7.33 (s, 2H), 5.36 (dd, J= 17.1, 1.5 Hz, 1H), 5.14 (d, J= 17.1 Hz, 1H), 4.13 (dd, J= 8.8, 5.2

Hz, 1H), 3.93 - 3.77 (m, 1H), 3.71 - 3.60 (m, 1H), 2.45 - 2.34 (m, 1H), 2.27 - 2.00 (m, 4H), 1.94-1.50 (m, 5H), 1.34-1.13 (m, 1H), 1.08-0.95 (m, 1H), 0.71-0.62 (m, 1H); MS (ES+): 452.1 (M+l).

Scheme 103

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-((2,2dichlorocyclopropyl)methyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (103b)

Compound 103b was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyI)-2-azabicyclo[3.1.0]hexane-320 carboxylic acid (95a) (120 mg, 0.397 mmol) in DMF (1 mL) using (2,2dichlorocyclopropyl)methanamine (103a) (55.6 mg, 0.397 mmol; CAS # 226549-07-1), HATU (226 mg, 0.595 mmol), DIPEA (0.207 mL, 1.191 mmol) and stirring atRT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), using DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyI)-N-((2,2dichlorocyclopropyl)methyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (103b) (67 mg, 40% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/₆) δ 8.39 (s, 1H), 8.33 (s, 1H),

222

8.20 - 8.10 (m, 1H), 5.49 (d, J= 17.1 Hz, 1H), 5.23 (d, J= 17.0 Hz, 1H), 4.28 - 4.19 (m, 1H), 3.72 - 3.63 (m, 1H), 3.30 - 3.11 (m, 2H), 2.30 - 2.02 (m, 2H), 1.93 - 1.75 (m, 2H), 1.68 . (ddd, J= 10.8, 7.2, 3.6 Hz, 1H), 1.34 (t, J= 7.5 Hz, 1H), 1.05 (dt, J= 9.6, 5.3 Hz, 1H), 0.74 0.62 (m, 1H); MS (ES+) 424.1 (M+l).

Scheme 104

Préparation of (2S,4R)-l-(2-(6-amino-9H-purin-9-yI)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (104a)

Compound 104a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (lOOmg, 0.518 mmol) in DMF (2 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (7a) (142 mg, 0.518 mmol), HATU (295 mg, 0.777 mmol), DIPEA (0.271 mL, 1.553 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [Cl8 column (30 g), eluting with ACN in water (containing 0.1% HCl) from

0-100%] (2S,4R)-l-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (104a) (57 mg, 24% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSOW) δ 8.64 (t, J= 5.9 Hz, 1H), 8.40 - 8.33 (m, 1H), 8.33 - 8.25 (m, 1H), 7.52 - 7.34 (m, 1H), 7.29 - 7.14 (m, 1H), 7.06 (td, J= 7.9, 1.1 Hz, 1H), 5.63 - 5.06 (m, 3H), 4.92 - 3.72 (m, 5H), 2.21 - 1.86 (m, 1H); MS (ES+) 450.1 (M+l); Analysis calculated for C19H18CIF2N7O2.2.5H2O.HCI: C, 42.95; H, 4.55; Cl, 13.34; N, 18.45; Found: C, 42.80; H, 4.26; Cl, 13.15; N, 18.25.

Scheme 105

223

Préparation of (S)-3-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2fluorobenzyl)thiazolidine-2-carboxamide (105b)

Compound 105b was prepared according to the procedure reported in step-3 of scheme-1, 5 from 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (100 mg, 0.518 mmol) in DMF (2 mL) using (S)-N-(3-chloro-2-fluorobenzyl)thiazolidine-2-carboxamide (105a) [142 mg, 0.518 mmol; prepared according to the procedure reported by Altmann, Eva et al., PCT Int. Appl. (2012), WO 2012093101 Al 20120712], HATU (295 mg, 0.777 mmol), DIPEA (0.271 mL, 1.553 mmol) and stirring at RT for 16 h. The solid obtained was collected by filtration washed with water, ethyl acetate and dried to yield (S)-3-(2-(6-amino-9H-purin-9-yl)acetyl)N-(3-chloro-2-fluorobenzyl)thiazolidine-2-carboxamide (105b) (75mg, 32% yield) as a white solid; ’H NMR (300 MHz, DMSO-î7₆) δ 8.62 (t, J= 5.9 Hz, 1H), 8.15 - 8.09 (m, 1H), 8.02 (s, 1H), 7.46 (td, J= 7.6, 1.8 Hz, 1H), 7.32-7.19 (m, 3H), 7.12 (td, J= 7.8, 1.0 Hz, 1H), 5.44 (s, 1H), 5.32 - 5.11 (m, 2H), 4.52 - 3.74 (m, 4H), 3.40 - 3.09 (m, 2H); MS (ES+): 450.1 (M+l); (ES-): 448.1 (M-l).

Scheme 106

95a

HATU, DIPEA

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(5-hydroxy-3,3dimethylcyclohexyl)-2-azabicyclo[3.1,0]hexane-3-carboxamide (106b)

224

Compound 106b was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3carboxylic acid (95a) (120 mg, 0.397 mmol) in DMF (1 mL) using racemic-(lR,5S)-5amino-3,3-dimethylcyclohexan-ol (106a) (56.9 mg, 0.397 mmol); CAS # 1529781-98-3),

HATU (189 mg, 0.496 mmol), DIPEA (0.207 mL, 1.191 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [Cl8 column (150 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(5-hydroxy-3,310 dimethylcyclohexyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (106b) (40mg, 24% yield) (racemic mixture) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ê) δ 8.47 (s, 1H), 8.42 (s, 1H), 7.65 - 7.54 (m, 1H), 5.53 (d, J= 17.1 Hz, 1H), 5.24 (d, J= 17.0 Hz, 1H), 4.21 4.07 (m, 1H), 3.89 - 3.22 (m, 3H), 2.24 - 1.99 (m, 2H), 1.94 - 1.76 (m, 2H), 1.60 - 1.42 (m, 1H), 1.42 - 1.27 (m, 1H), 1.08 - 0.89 (m, 4H), 0.89 (s, 3H), 0.85 (s, 3H), 0.73 - 0.57 (m,

1H); Analysis calculated for C21H29N7O3.I.5H2O.2.75HCI.: C, 45.46; H, 6.31; Cl, 17.57; N,

17.67; found: C, 45.75; H, 6.68; Cl, 17.63; N, 17.68.

Scheme 107

95a

107b

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3 (trifluoromethoxy)phenyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (107b)

Compound 107b was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (95a) (120 mg, 0.397 mmol) in DMF (1 mL) using 3-(trifluoromethoxy)aniline (107a) (55.6 mg, 0.397 mmol; CAS # 1535-73-5), HATU (226 mg, 0.595 mmol), DIPEA (0.207 mL, 25 1.191 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [Cl8 column (150 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-amino

225

9H-purin-9-yl)acetyl)-N-(3-(trifluoromethoxy)phenyl)-2-azabicyclo[3.1.0]hexane-3carboxamide (107b) (97 mg, 53% yield) HCl sait as a white solid; *HNMR (300 MHz, DMSOdô) δ 10.20 (s, 1H), 8.12 (s, 1H), 8.06 (s, 1H), 7.77 - 7.74 (m, 1H), 7.54 - 7.36 (m, 2H), 7.19 (s, 2H), 7.06 - 7.00 (m, 1H), 5.41 (d, J= 17.2 Hz, 1H), 5.18 (d, J= 17.1 Hz, 1H), 4.33 (dd, J = 8.8, 5.6 Hz, 1H), 3.76 (ddd, J= 7.2, 5.4, 2.4 Hz, 1H), 2.39 - 2.16 (m, 2H), 2.00 - 1.80 (m, 1H), 1.05 (dt, 8.7, 5.4 Hz, 1H), 0.74 (td, J= 5.1, 2.4 Hz, 1H); ¹⁹F NMR (282 MHz, DMSOdd) δ -56.65; MS (ES+): 462.1 (M+l).

Scheme 108

95a

108b

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(2-fluoro-3methoxyphenyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (108b)

Compound 108b was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-2-azabicyclo[3.1,0]hexane-3carboxylic acid (95a) (120 mg, 0.397 mmol) in DMF (1 mL) using 2-fluoro-3methoxyaniline (108a) (67.2 mg, 0.476 mmol; CAS # 801282-00-8), HATU (226 mg, 0.595 mmol), DIPEA (0.207 mL, 1.191 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [C 18 column (150 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(2-fluoro-3-methoxyphenyl)-2azabicyclo[3.1.0]hexane-3-carboxamide (108b) (65 mg, 39% yield) HCl sait as a white solid;

Ή NMR (300 MHz, DMSO-rf₆) δ 9.69 (s, 1H), 8.48 (s, 1H), 8.44 (s, 1H), 7.40 - 7.33 (m, 1H), 7.05 (td, J= 8.3, 1.8 Hz, 1H), 6.93 (td, J= 8.2, 1.6 Hz, 1H), 5.58 (d, J= 17.2 Hz, 1H), 5.29 (d, J= 17.1 Hz, 1H), 4.52 (dd, J= 8.5, 5.5 Hz, 1H), 3.82 (s, 3H), 3.80 - 3.65 (m, 1H), 2.37-2.16 (m, 2H), 1.97- 1.84 (m, 1H), 1.07 (dt, J= 9.2, 5.3 Hz, 1H), 0.72 (td, J= 5.1, 2.4 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-ùfo) δ -147.09 (t, J = 7.3 Hz); MS (ES+): 426.1 (M+l).

Scheme 109

226

Préparation of (lS,3S,5S)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)2-azabicyclo[3.1,0]hexane-3-carboxamide (109e)

Step-1: Préparation of tert-butyl (lS,3S,5S)-3-((3-chloro-2-fluorobenzyl)carbamoyl)-25 azabicyclo[3.1.0]hexane-2-carboxylate (109c)

Compound 109c was prepared according to the procedure reported in step-3 of scheme-1, from (lS,3S,5S)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (109a) (0.5 g, 2.2 mmol; CAS # 197142-36-2) using (3-chloro-2-fluorophenyl)methanamine (109b) (0.386 g, 2.420 mmol; CAS # 72235-55-3), (HATU) (1.255 g, 3.30 mmol), DIPEA (1.150 mL, 6.60 mmol) in DMF (5 mL) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with MeOH in DCM 0-10%] tert-butyl ( 1 S,3S,5S)-3-((3-chloro-2-fluorobenzyl)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (109c) (0.8 g, 99% yield) as a pale-yellow gel; MS (ES+): 391.1 (M+Na).

Step-2: Préparation of (lS,3S,5S)-N-(3-chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3carboxamide (109d)

Compound 109d was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl (1 S,3S,5S)-3-((3-chloro-2-fluorobenzyl)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (109c) (0.75 g, 2.033 mmol) using TFA (0.627 mL, 20 8.13 mmol) in DCM (6 mL) and stirring ovemight at room température. This gave after workup and purification by reverse phase column chromatography [Cl8 (140 g), eluting with ACN in water (containing 0.1% HCl) from 0 to 100%] (lS,3S,5S)-N-(3-chloro-2-

227 fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (109d); Ή NMR (300 MHz, DMSO-dô) δ 10.33 (s, 1H), 9.11 (t, J= 5.7 Hz, 1H), 8.79 (d, J= 9.1 Hz, 1H), 7.52 (ddd, J= 7.9, 7.1, 1.8 Hz, 1H), 7.33 (ddd, J= 7.8, 6.7, 1.8 Hz, 1H), 7.22 (td, J= 7.8, 1.1 Hz, 1H), 4.54 (s, 1H), 4.39 (d, J= 5.6 Hz, 2H), 3.29 (s, 1H), 2.55 (s, 1H), 2.10 (dd, J= 13.7, 3.1 Hz, 1H), 1.75 (dq, J= 9.0, 5.4 Hz, 1H), 0.90 - 0.76 (m, 1H), 0.65 (ddd, J= 7.1, 5.0, 2.6 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -121.11; MS (ES+): 269.1 (M+l).

Step-3: Préparation of (lS,3S,5S)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (109e)

Compound 109e was prepared according to the procedure reported in step-3 of scheme-1, from (lS,3S,5S)-N-(3-chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (109d) (139 mg, 0.518 mmol) in DMF (2 mL) using 2-(6-amino-9H-purin-9-yl)acetic acid (lOOmg, 0.518 mmol) (67a), HATU (295 mg, 0.777 mmol), DIPEA (0.271 mL, 1.553 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [Cl 8 column (150 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lS,3S,5S)-2-(2-(6-amino-9Hpurin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (109e) (71mg, 31% yield) as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.49 (t, J= 5.8 Hz, 1H), 8.42 (s, 1H), 8.35 (s, 1H), 7.52-7.37 (m, 1H), 7.29-7.18 (m, 1H), 7.10 (td, J= 7.9, 1.0 Hz, 1H), 5.52 (d, J= 17.0 Hz, 1H), 5.27 (d,J=17.0Hz, 1H), 4.67 (dd, J= 11.2, 3.4 Hz, 1H), 4.49-4.10 (m, 2H), 3.86 - 3.69 (m, 1H), 2.63-2.51 (m, 1H), 1.89- 1.67 (m, 2H), 1.16 (td, J= 5.1, 2.4 Hz, 1H), 0.89 - 0.78 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î/î) δ 121.60; MS (ES+): 444.1 (M+l).

Scheme 110

228

Préparation of (2S,4R)-l-(2-(6-amino-8-bromo-9H-purin-9-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (HOd)

Step-1: Préparation of ZerZ-butyl 2-(6-amino-8-bromo-9H-purin-9-yl)acetate (110b)

Compound 110b was prepared according to the procedure reported in step-1 of scheme-1, from 8-bromo-9H-purin-6-amine (110a) (1 g, 4.67 mmol; CAS#6974-78-3) in DMF (10 mL) using ZerZ-butyl 2-bromoacetate and K2CO3 (0.775 g, 5.61 mmol). This gave after workup and recrystallization with ethyl acetate ZerZ-butyl 2-(6-amino-8-bromo-9H-purin-9-yl)acetate (110b) (550 mg, 36% yield); Ή NMR (300 MHz, DMSO-î/₆) δ 8.33 (s, 1H), 8.31 (s, 1H), 8.15 (s, 1H), 5.04 (s, 2H), 1.43 (s, 9H).

Step-2: Préparation of 2-(6-amino-8-bromo-9H-purin-9-yl)acetic acid (110c)

Compound 110c was prepared according to the procedure reported in step-4 of scheme-17, from ZerZ-butyl 2-(6-amino-8-bromo-9H-purin-9-yl)acetate (110b) (0.5 g, 1.524 mmol) in THF (1.676 mL) and MeOH (1.676 mL) using 2N sodium hydroxi de (0.838 mL, 1.676 mmol) and stirring at RT for 45 min. This gave after workup 2-(6-amino-8-bromo-9H-purin9-yl)acetic acid (110c) (346 mg, 83% yield); Ή NMR (300 MHz, DMSO-ity δ 8.31 (s, 1H), 5.04 (s, 2H).

Step-3: Préparation of (2S,4R)-l-(2-(6-amino-8-bromo-9H-purin-9-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoropynOlidine-2-carboxamide (HOd)

Compound HOd was prepared according to the procedure reported in step-3 of scheme-1, from 2-(6-amino-8-bromo-9H-purin-9-yl)acetic acid (110c) (141mg, 0.518 mmol) in DMF (2 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (7a) (142 mg,

229

0.518 mmol), HATU (296 mg, 0.777 mmol), DIPEA (0.271 mL, 1.555 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [Cl8 column (150 g), eluting with ACN in water 5 (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(6-amino-8-bromo-9H-purin-9-yl)acetyl)N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (HOd) (123 mg, 45% yield) as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 8.63 (t, J= 5.9 Hz, 1H), 8.38 (s, 2H), 8.29 (s, 1H), 7.52 - 7.32 (m, 1H), 7.28 - 7.13 (m, 1H), 7.07 - 7.00 (m, 1H), 5.64 - 5.12 (m, 3H), 4.94 - 3.75 (m, 5H), 2.57 - 1.93 (m, 2H); ¹⁹F NMR (282 MHz, DMSO-7₆) δ -121.69, -173.83 10 --179.95; MS (ES+): 528.0 (M+l).

Scheme 111

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(2-fluoro-3-methylbut-2en-l-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (111b)

Compound 111b was prepared according to the procedure reported in step-3 of scheme-1, from 2-fluoro-3-methylbut-2-en-l-amine (111a) (61.4 mg, 0.595 mmol; prepared according to the procedure reported by Flohr, Stefanie et al; in PCT Int. AppL, 2014002052, 03 Jan 2014) in DMF (1 mL) using (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-2azabicyclo[3.1.0]hexane-3-carboxylic acid (95a) (120 mg, 0.397 mmol), HATU (226 mg,

0.595 mmol), DIPEA (0.277 mL, 1.588 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [C18 column (150 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-amino9H-purin-9-yl)acetyl)-N-(2-fluoro-3-methylbut-2-en-l-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (111b) (35 mg, 23% yield) HCl sait as a white solid; Ή NMR (300 MHz,

OMSO-de) δ 8.49 (s, 1H), 8.42 (s, 1H), 8.17 (t, J= 5.6 Hz, 1H), 5.53 (d, J= 17.1 Hz, 1H), 5.25 (d, J= 17.1 Hz, 1H), 4.23 (dd, J= 9.0, 4.6 Hz, 1H), 3.86 (dt, J= 21.9, 5.6 Hz, 2H), 3.70 -3.58 (m, 1H), 2.32-2.01 (m, 2H), 1.92- 1.78 (m, 1H), 1.63 (d, 7=2.9 Hz, 3H), 1.57 (d, J

230

= 3.3 Hz, 3H), 1.13 - 0.97 (m, 1H), 0.75 - 0.61 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-^) δ 68.80; MS (ES+): 388.2 (M+l); (ES-): 386.2 (M-l).

Scheme 112

HATU, DIPEA

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (112b)

Compound 112b was prepared according to the procedure reported in step-3 of scheme-1, from 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (47.8 mg, 0.247 mmol) in DMF (2 mL) using (lR,3S,5R)-N-(6-bromopyrazin-2-yl)-2-azabicyclo[3.1,0]hexane-3-carboxamide (112a) [70 mg, 0.247 mmol; prepared according to the procedure reported by Wiles, Jason A et al., PCT Int. Appl. (2017), WO 2017035355 Al 20170302], HATU (141 mg, 0.371 mmol), DIPEA (0.172 mL, 0.989 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [Cl8 column (150 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6amino-9H-purin-9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (112b) (14 mg, 12% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO<Z₆) δ 11.17 (s, 1H), 9.26 (d, J= 0.6 Hz, 1H), 8.55 (d, J= 0.6 Hz, 1H), 8.41 (s, 1H), 8.36 (s, 1H), 5.57 (d, J= 17.2 Hz, 1H), 5.27 (d, J= 17.1 Hz, 1H), 4.48 (dd, J= 8.9, 5.5 Hz, 1H), 3.86 - 3.70 (m, 1H), 2.39 - 2.15 (m, 2H), 1.97 - 1.81 (m, 1H), 1.14 - 0.94 (m, 1H), 0.73 (td, J = 5.1, 2.4 Hz, 1H); MS (ES+): 458.0, 460.0 (M+l).

Scheme 113

231

Préparation of (lR,3S,5R)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(6-bromo-3methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (113a)

Compound 113a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (65 mg, 0.337 mmol) in DMF (1.5 mL) using (lR,3S,5R)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1,0]hexane-3carboxamide (10a) [115 mg, 0.370 mmol), HATU (192 mg, 0.505 mmol), DIPEA (0.234 mL, 1.346 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [Cl 8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-amino9H-purin-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (113a) (39 mg, 24% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.35 (s, 1H, D₂O exchangeable), 9.63 (s, 1H, D₂O exchangeable), 8.88 (s, 1H, D₂O exchangeable), 8.51 (s, 1H), 8.45 (s, 1H), 7.64 (d, J= 8.0 Hz, 1H), 7.45 (d, J= 7.9 Hz, 1H), 5.55 (d, J= 17.1 Hz, 1H), 5.25 (d, J= 17.1 Hz, 1H), 4.41 (dd, J= 9.2, 5.1 Hz, 1H), 3.54 (dd, J= 5.5, 2.4 Hz, 1H), 2.57 - 2.53 (m, 1H), 2.06 (s, 3H), 2.01 (d, J=5.1Hz, 1H), 1.30 (s, 3H), 1.02 (t,J= 5.4 Hz, 1H), 0.92 (dd, J= 5.2, 2.4 Hz, 1H); MS (ES+): 485.1 (M+l), 507.0 (M+Na); (ES-): 483.0 (M-l); Calculated for C₂₀H₂|BrN₈O₂.1.25(HCl).2.75(H₂O): C, 41.38; H, 4.82; Cl, 7.63; N, 19.30 found: C, 41.65; H, 4.62; Cl, 7.61; N, 19.04.

Scheme 114

232

Préparation of (2S,4R)-l-(2-(4-amino-5-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (1141)

Step-1: Préparation of ZerZ-butyl 2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetate (114b)

Compound 114b was prepared according to the procedure reported in step-1 of scheme-1, from 5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (114a) (2 g, 9.39 mmol) in DMF (50 mL) using ZerZ-butyl 2-bromoacetate (1.387 mL, 9.39 mmol), CS2CO3 (3.67 g, 11.27 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with EtOAc in Hexane from 0-50%] ZerZ-butyl 2(4-ammo-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (114b) (2.68 g, 87% yield) as a light pink solid; MS (ES+): 327.00 (M+l).

Step-2: Préparation of ZerZ-butyl 2-(4-amino-5-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetate (114d)

Compound 114d was prepared according to the procedure reported in step-1 of scheme-59, from ZerZ-butyl 2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (114b) in dioxane (10 mL) using pyridin-3-ylboronic acid (114c) (113 mg, 0.917 mmol; CAS # 169225-7), bis(triphenylphosphine)palladium(II) chloride (97 mg, 0.138 mmol) a solution of potassium carbonate (380 mg, 2.75 mmol) in water (1.250 mL) and heating at 100 °C for 5 h. This gave after workup and purification by chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0-50%] ZerZ-butyl 2-(4-amino-5-(pyridin-3-yl)-7H-pyrrolo[2,3-

233

d]pyrimidin-7-yl)acetate (114d) (188 mg, 63% yield) as a yellow solid; *H NMR (300 MHz, DMSO-<7₆) δ 8.71 - 8.62 (m, 1H), 8.54 (dd,J=4.9, 1.6 Hz, 1H), 8.16 (s, 1H), 7.90-7.75 (m, 1H), 7.48 (dd, J= 7.9, 4.8 Hz, 1H), 7.43 (s, 1H), 6.24 (s, 2H), 4.95 (s, 2H), 1.43 (s, 9H); MS (ES+): 326.1(M+1).

Step-3: Préparation of 2-(4-amino-5-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (114e)

Compound 114e was prepared according to the procedure reported in step-4 of scheme-17, from tert-butyl 2-(4-amino-5-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (114d) in THF/MeOH (2 mL; ratio 1:1) using 2N sodium hydroxide (0.318 mL, 0.636 mmol) and stirring at RT for 2 h. This gave after workup 2-(4-amino-5-(pyridin-3-yl)-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetic acid (114e) (114 mg, 77% yield) as a yellow solid; 'H NMR (300 MHz, DMSO-î/ô) δ 8.68 (dd, J= 2.3, 0.9 Hz, 1H), 8.54 (dd, J=4.8, 1.6 Hz, 1H), 8.16 (s, 1H), 7.84 (dt, J= 7.9, 1.9 Hz, 1H), 7.53 - 7.39 (m, 2H), 6.24 (s, 2H), 4.95 (s, 2H); MS (ES+): 270.10 (M+l).

Step-4: Préparation of (2S,4R)-l-(2-(4-amino-5-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (1141)

Compound 114f was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-5-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (114e) (lOOmg, 0.371 mmol) in DMF (5 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-420 fluoropyrrolidine-2-carboxamide (7a) (122 mg, 0.446 mmol), HATU (212 mg, 0.557 mmol), DIPEA (0.259 mL, 1.486 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), using DMA80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [C 18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-125 (2-(4-amino-5-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (114f) (11 Img, 57% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-t/ό) (a mixture of two rotamers) δ 8.91 (s, J= 2.1 Hz, 1H), 8.82 (d, J= 5.6 Hz, 1H), 9.16 (t, J=5.7 Hz) and 8.69 (t, J= 5.9 Hz) (2t,lH) (D₂O exchangeable), 8.57 - 8.18 (m, 2H), 7.98 (dd, J= 8.1, 5.5 Hz, 1H), 7.77 (s, 1H), 7.46 - 7.29 (m, 1H), 7.21-7.06 (m, 1H), 6.96 (td, J= 7.9, 1.1 Hz, 1H), 5.57-5.13 (m, 3H), 4.91 - 3.72 (m, 5H), 3.82 - 3.24 (s, 2H), 2.54 - 2.35 (m, 1H), 2.13 - 1.83 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î7₆) δ-121.31, -121.62, -176.11; MS (ES+): 526.2 (M+l); (ES-): 524.1(M-1), 560.1

234 (M+Cl); Analysis Calculated for C25H22CIF2N7O2.2HCI.3.5H2O: C, 45.36; H, 4.72; N, 14.81;

found: C, 45.43; H, 4.65; N, 14.67.

Scheme 115

Préparation of (2S,4R)-l-(2-(4-amino-5-(3-(aminomethyl)phenyl)-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (115e)

Step-1: Préparation of tert-butyl 2-(4-amino-5-(3-(((teributoxycarbonyl)amino)methyl)phenyl)-7H-pynOlo[2,3-d]pyrimidin-7-yl)acetate (115b)

Compound 115b was prepared according to the procedure reported in step-lof scheme-59, from tert-butyl 2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (114b) (300 mg, 0.917 mmol) in dioxane (10 mL) using te/7-butyl 3-(4,4,5,5-tetramethyl-l,3,2dioxaborolan-2-yl)benzylcarbamate (115a) (306 mg, 0.917 mmol), bis(triphenylphosphine)palladium(II) chloride (97 mg, 0.138 mmol) a solution of potassium carbonate (380 mg, 2.75 mmol) in water (1.250 mL) and heating at 100 °C for 4 h. This gave after workup and purification by flash column chromatography [silica gel (24g), eluting with DMA-80 in DCM from 0-50%] tert-butyl 2-(4-amino-5-(3-(((tertbutoxycarbonyl)amino)methyl)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (115b) (240

235 mg, 58% yield) as a clear gel; Ή NMR (300 MHz, DMSO-7₆) δ 8.14 (s, 1H), 7.43 (t, J= 7.5 Hz, 2H), 7.37 - 7.19 (m, 4H), 6.12 (s, 2H), 4.94 (s, 2H), 4.19 (d, J= 6.1 Hz, 2H), 1.43 (s, 9H), 1.39 (s, 9H); MS (ES+): 454.2 (M+l).

Step-2: Préparation of 2-(4-amino-5-(3-(((toV-butoxycarbonyl)amino)methyl)phenyl)-7Hpyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (115c)

Compound 115c was prepared according to the procedure reported in step-4 of scheme-17, from tert-butyl 2-(4-amino-5-(3-(((te/7-butoxycarbonyl)amino)methyl)phenyl)-7Hpyrrolo[2,3-d]pyrimidin-7-yl)acetate (115b) (230 mg, 0.507 mmol) in THF/MeOH (2 mL; ratio 1:1) using 2N sodium hydroxide (0.292 mL, 0.583 mmol) and stirring for 1.5 h at RT. This gave after workup 2-(4-amino-5-(3-(((tert-butoxycarbonyl)amino)methyl)phenyl)-7Hpyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (115c) (137 mg, 68% yield) as apink solid; 'H NMR (300 MHz, DMSO-î/ô) δ 8.13 (s, 1H), 7.43 (t, J= 7.6 Hz, 2H), 7.37 - 7.14 (m, 4H), 6.11 (s, 2H), 4.93 (s, 2H), 4.19 (d, J = 6.1 Hz, 2H), 1.39 (s, 9H); MS (ES+): 398.20 (M+l).

Step-3: Préparation of ZerZ-butyl 3-(4-amino-7-(2-((2S,4R)-2-((3-chloro-2fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l-yI)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidin-5yl)benzylcarbamate (115d)

Compound 115d was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-5-(3-(((terf-butoxycarbonyl)amino)methyl)phenyl)-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetic acid (115c) (100 mg, 0.252 mmol) in DMF (5 mL) using (2S,4R)-N(3-chloro-2-fluorobenzyl)-4-fluoropynOlidine-2-carboxamide (7a) (83 mg, 0.302 mmol), HATU (144 mg, 0.377 mmol), DIPEA (0.175 mL, 1.006 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 50 %] ZerZ-butyl 3-(4-amino-7-(2-((2S,4R)-2-((3chloro-2-fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l-yl)-2-oxoethyl)-7H-pyrrolo[2,3d]pyrimidin-5-yl)benzylcarbamate (115d) (156 mg, 95% yield) as a white solid; MS (ES+): 654.3 (M+l).

Step-4: Préparation of (2S,4R)-l-(2-(4-amino-5-(3-(aminomethyl)phenyl)-7H-pynOlo[2,3d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropynOlidine-2-carboxamide (115e)

Compound 115e was prepared according to the procedure reported in step-2 of scheme-1, from ZerAbutyl 3-(4-amino-7-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropynOlidin-l-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)benzylcarbamate (115d)

236 (150mg, 0.229 mmol) in DCM (3 mL) using TFA (0.353 mL, 4.59 mmol) and stirring at RT for 70 min. This gave after workup and purification by reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-5-(3-(aminomethyl)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (115e) (92 mg, 72%yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-riô) (a mixture of two rotamers) δ 9.19 (t, J= 5.8 Hz) and 8.74 (t, J= 5.9 Hz) (2t, 1H), 8.53 - 8.40 (m, 4H, 3H D₂O exchangeable), 7.66 - 7.35 (m, 6H), 7.30 - 7.14 (m, 1H), 7.02 (t, J= 7.9 Hz, 1H), 5.66 -5.15 (m, 3H), 4.96 3.88 (m, 7H), 2.59 - 2.41 (m, 1H), 2.23 - 1.89 (m, 1H); ¹⁹F NMR (282 MHz, DMSO) δ 121.29, -121.63, -175.96; MS (ES+): 554.2(M+1); (ES-): 552.2(M-1), 588.1 (M+Cl);

Analysis calculated for C₂₇H₂₆C1F₂N₇O₂.2HC1.3H₂O: C, 47.62; H, 5.03; Cl, 15.62; N, 14.40; Found; C, 47.74; H, 4.94; Cl, 15.37; N, 14.30.

Scheme 116

Préparation of (2S,4R)-l-(2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (116b)

Step-1: Préparation of 2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (116a)

Compound 116a was prepared according to the procedure reported in step-4 of scheme-17, from te/7-butyl 2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (114b) (200 mg, 0.611 mmol) in THF/MeOH (2.4 mL; ratio 1:1) using 2N sodium hydroxide (0.367 mL, 0.734 mmol) and stirring for 1 h at RT. This gave after workup 2-(4-amino-5-bromo-7Hpyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (116a) (138 mg, 83% yield) as a pale purple solid; ‘H NMR (300 MHz, DMSO-i/e) δ 13.11 (s, 1H), 8.08 (s, 1H), 7.41 (s, 1H), 6.76 (s, 2H), 4.87 (s, 2H).

237

Step-2: Préparation of (2S,4R)-l-(2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (116b)

Compound 116b was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-5-bromo-7H-pynOlo[2,3-d]pyrimidin-7-yl)acetic acid (116a) (50 mg, 0.184 mmol) in DMF (5 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2carboxamide (7a) (51 mg, 0.184 mmol), HATU (105 mg, 0.277 mmol), DIPEA (0.129 mL, 0.738 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-5bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (116b) (29 mg, 30% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-ίΖό) (a mixture of two rotamers) δ 9.04 (t, J= 5.7 Hz) and 8.63 (t, J= 5.8 Hz) (2t,lH), 8.28 (s) and 8.26 (s) (2s, 1H), 7.83 (s, 3H, D₂O exchangeable), 7.53 (s, 1H), 7.49 - 7.34 (m, 1H), 7.29 - 7.13 (m, 1H), 7.08 (td, J= 7.9, 1.1 Hz, 1H), 5.47 (d, J= 52.8 Hz, 1H), 5.31 - 5.02 (m, 2H), 4.84 - 3.73 (m, 5H), 2.43 (m, 1H), 2.19 - 1.85 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-îZô) δ -121.32, -121.62, -176.08. MS (ES+): 527.0 (M+l); (ES-): 525.0 (M-l); Analysis calculated for C₂oHi8BrClF₂N60₂.HC1.1.5H₂0: C, 40.63; H, 3.75; N, 14.21; Found: C, 40.70; H, 3.54; N, 13.95.

Scheme 117

238

Préparation of (2S,4R)-l-(2-(4-amino-5-(2-methylpyrimidin-5-yl)-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (117d)

Step-1: Préparation of ZerZ-butyl 2-(4-amino-5-(2-methylpyrimidin-5-yl)-7H-pynOlo[2,3d]pyrimidin-7-yl)acetate (117b)

Compound 117b was prepared according to the procedure reported in step-1 of scheme-59, from ZerZ-butyl 2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (114b) (1 g, 3.06 mmol) in dioxane (30 mL) using 2-methyl-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-270 yl)pyrimidine (117a) (673 mg, 3.06 mmol; CAS # 1052686-67-5), bis(triphenylphosphine)palladium(II) chloride (322 mg, 0.458 mmol) a solution of potassium carbonate (1.267 g, 9.17 mmol) in water (3.75 mL) and heating at 100 °C for 3 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0-50%] ZerZ-butyl 2-(4-amino-5-(2-methylpyrimidin-5-yl)-7H15 pyrrolo[2,3-d]pyrimidin-7-yl)acetate (117b) (375 mg, 36% yield) as a white solid; Ή NMR (300 MHz, DMSO-î/ô) δ 8.72 (s, 2H), 8.16 (s, 1H), 7.47 (s, 1H), 6.43 (s, 2H), 4.95 (s, 2H), 2.66 (s, 3H), 1.43 (s, 9H); MS (ES+): 341.1 (M+l).

Step-2: Préparation of 2-(4-amino-5-(2-methylpyrimidin-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetic acid (117c)

239

Compound 117c was prepared according to the procedure reported in step-4 of scheme-17, from tert-butyl 2-(4-amino-5-(2-methylpyrimidin-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetate (117b) (350 mg, 1.028 mmol) in THF/MeOH (4 mL; ratio 1:1) using 2N sodium hydroxide (0.6 mL, 1.20 mmol) and stirring for 3 h at RT. This gave after workup 2-(4amino-5-(2-methylpyrimidin-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (117c) (167 mg, 57% yield) as a white solid; Ή NMR (300 MHz, DMSO-î7₆) δ 8.72 (s, 2H), 8.16 (s, 1H), 7.48 (s, 1H), 6.43 (s, 2H), 4.96 (s, 2H), 2.66 (s, 3H); MS (ES+): 285.1 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-5-(2-methylpyrimidin-5-yl)-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (117d)

Compound 117d was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-5-(2-methylpyrimidin-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (117c) (50 mg, 0.176 mmol) in DMF (3 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (7a) (48.3 mg, 0.176 mmol), HATU (100 mg, 0.264 mmol), DIPEA (0.123 mL, 0.704 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-5-(2-methylpyrimidin-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (117d) (67 mg, 70% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 9.22 and 8.75 (2t, J= 5.9 Hz, 1H, D₂O exchangeable), 8.80 (s, 2H), 8.52 (s, 1H), 8.40 (s, 2H, D₂O exchangeable), 7.73 and 7.72 (2s, 1H), 7.49-7.34 (m, 1H), 7.29-7.13 (m, 1H), 7.02 (td,J=7.9, 1.1 Hz, 1H), 5.63-4.68 (m, 3H), 4.52 - 3.76 (m, 5H), 2.72 (s, 3H), 2.60-2.19 (m, 1H), 2.18-1.93 (m, 1H). Ή NMR (300 MHz, DMSO-îZ₆/D₂O) δ 8.80 (s, 2H), 8.48 and 8.46 (2s, 1H), 7.70 and 7.68 (2s, 1H), 7.49-7.33 (m, 1H), 7.28 - 7.13 (m, 1H), 7.04 (td, .7=7.9, 1.1 Hz, 1H), 5.63 - 5.15 (m, 3H), 4.52 - 3.84 (m, 5H), 2.73 (s, 3H), 2.52 - 2.25 (m, 1H), 2.23 - 1.90 (m, 1H); ¹⁹F NMR (282 MHz, DMSO) δ -121.30, -121.61, -176.09; MS (ES+): 541.2 (M+l); (ES-): 539.2 (ΜΙ); Analysis calculated for C₂₅H₂₃C1F₂N₈O₂.1.65HC1.3.75H₂O: C, 44.91; H, 4.85; Cl, 14.05; N, 16.76; Found: C, 45.01; H, 4.68; Cl, 14.10; N, 16.44.

Scheme 118

240

Préparation of (2S,4R)-l-(2-(4-amino-5-(2-methylpyrimidin-5-yl)-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-4-fluoropynOlidine-2carboxamide (118a)

Compound 118a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-5-(2-methylpyrimidin-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (117c) (50 mg, 0.176 mmol) in DMF (3 mL) using (2S,4R)-N-(6-bromo-3-methylpyridin-2yl)-4-fluoropyrrolidine-2-carboxamide (3a) (53 mg, 0.176 mmol), HATU (100 mg, 0.264 mmol) and DIPEA (0.123 mL, 0.704 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-5-(2-methylpyrimidin-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (118a) (37 mg, 37% yield) HCl sait as a pale yellow solid; Ή NMR (300 MHz, DMSO-7₆) δ 10.92 and 10.50 (2s, 1H, D₂O exchangeable), 8.78 (s, 2H), 8.50 (s, 1H), 8.32 (s, 2H, D2O exchangeable), 7.78 and 7.71 (2s, 1H), 7.64 - 7.57 (m, 1H), 7.52 and 7.43 (2d, J= 7.9 Hz, 1H), 5.65-5.21 (m, 3H),4.61 (t, 7= 8.5 Hz, 1H), 4.21 (dd, J=21.7, 12.5 Hz, 1H), 3.94 (dd, 7= 38.0, 12.6 Hz, 1H), 2.70 (s, 3H), 2.67-2.60 (m, 1H), 2.34-2.06 (m, 1H), 2.03 (s, 3H); ¹⁹F

NMR (282 MHz, DMSO-7₆) δ -176.04. MS (ES+): 568.1 (M+l); (ES-): 566.1 (M-l).

Scheme 119

241

Préparation of (S)-l-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2fluorobenzyl)pyrrolidine-2-carboxamide (119a)

To a mixture of 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (50 mg, 0.259 mmol) and (S)-N(3-chloro-2-fluorobenzyl)pyrrolidine-2-carboxamide (76a) (66.4 mg, 0.259 mmol) in DMF (1 mL) was added lH-benzo[d][l,2,3]triazol-l-ol (HOBT) (17.49 mg, 0.129 mmol), Nl((ethylimino)methylene)-N3,N3-dimethylpropane-l,3-diamine hydrochloride (EDC) (99 mg, 0.518 mmol), DIPEA (0.135 mL, 0.777 mmol) and stirred at RT for 16 h. This mixture was diluted with ethyl acetate (120 mL), washed with water (2 x 30 mL), brine (30 mL), dried, filtered and concentrated under vacuum. The residue obtained was purified by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100 %] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] to afford (S)-l-(2-(6-amino-9H-purin-9yl)acetyl)-N-(3-chloro-2-fluorobenzyl)pyrrolidine-2-carboxamide (119a) (9 mg, 8% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.49 (t, J= 5.9 Hz, 1H), 8.36 (s, 1H), 8.25 (s, 1H), 7.53-7.32 (m, 1H), 7.26 - 7.15 (m, 1H), 7.09 (td, J= 7.9, 1.0 Hz, 1H), 5.30 -5.11 (m, 2H), 4.78 - 4.40 (m, 1H), 4.36 - 4.25 (m, 2H), 3.79-3.61 (m, 2H), 2.20 - 1.69 (m, 4H). ^,9FNMR(282 MHz, DMSO-î/₆) δ-121.70; MS (ES+): 432.1 (M+l); (ES-): 430.1 (M-l), 466.1 (M+Cl).

Scheme 120

242

Préparation of (S)-5-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-5azaspiro[2.4]heptane-6-carboxamide (120c)

Step-1: Préparation of (S)-tert-butyl 6-((3-chloro-2-fluorobenzyl)carbamoyl)-5azaspiro[2.4]heptane-5-carboxylate ( 120a)

Compound 120a was prepared according to the procedure reported in step-3 of scheme-1, from (S)-5-(ieri-butoxycarbonyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (53a) (0.5 g, 2.072 mmol) using (3-chloro-2-fluorophenyl)methanamine (109b) (0.331 mg, 2.072 mmol), (HATU) (1.182 g, 3.11 mmol), DIPEA (1.083 mL, 6.22 mmol) in DMF (5 mL) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with EtOAc/MeOH in Hexanes 0-100%] (S)-fôrt-butyl 6-((3-chloro-2fluorobenzyl)carbamoyl)-5-azaspiro[2.4]heptane-5-carboxylate (120a) as a pale-yellow gel (0.758 g, 96% yield); MS (ES+): 405.1 (M+Na).

Step-2: Préparation of (S)-N-(3-chloro-2-fluorobenzyl)-5-azaspiro[2.4]heptane-6carboxamide (120b)

Compound 120b was prepared according to the procedure reported in step-2 of scheme-1, from (S)-tert-butyl 6-((3-chloro-2-fluorobenzyl)carbamoyl)-5-azaspiro[2.4]heptane-5-carboxylate (120a) (0.758 g, 1.980 mmol) using TFA (0.763 mL, 9.90 mmol) in DCM (6 mL) and stirring ovemight at room température. This gave after workup and purification by reverse phase column chromatography [Cl8 (30 g), eluting with ACN in water (containing 0.1% HCl) from 0 to 100%] (S)-N-(3-chloro-2-fluorobenzyl)-5-azaspiro[2.4]heptane-6-carboxamide (120b)

243 (0.535 g, 96% yield); Ή NMR (300 MHz, DMSO-d₆) δ 9.09 (t, J= 5.7 Hz, 1H), 8.78 (s, 1H), 7.53 (ddd, J= 7.9, 7.2, 1.8 Hz, 1H), 7.34 (ddd, J= 7.7, 6.7, 1.8 Hz, 1H), 7.22 (td, J= 7.8, 1.1 Hz, 1H), 4.56 - 4.26 (m, 3H), 3.14 (s, 2H), 2.23 (dd, J= 13.0, 8.3 Hz, 1H), 1.90 (dd, J= 13.0, 7.3 Hz, 1H), 0.78 - 0.42 (m, 4H); ^I9FNMR (282 MHz, DMSO-îZ₆) δ -121.07; MS (ES+): 283.1 (M+Na).

Step-3: Préparation of (S)-5-(2-(6-amino-9H-purin-9-yl)acetyI)-N-(3-chloro-2-fluorobenzyl)5-azaspiro[2.4]heptane-6-carboxamide (120c)

Compound 120c was prepared according to the procedure reported in scheme-119, from (S)N-(3-chloro-2-fluorobenzyl)-5-azaspiro[2.4]heptane-6-carboxamide (120b) (73.2 mg, 0.259 mmol) in DMF (5 mL) using 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (50 mg, 0.259 mmol), HOBT (17.49 mg, 0.129 mmol), EDC (99 mg, 0.518 mmol), DIPEA (0.180 mL, 1.035 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100 %] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (S)-5-(2-(6-amino-9H-purin-9-yl)acetyl)N-(3-chloro-2-fluorobenzyl)-5-azaspiro[2.4]heptane-6-carboxamide (120c) (11 mg, 9% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 8.94 and 8.64 (2t, J= 6.0 Hz, 1H), 8.45 (s, 1H), 8.37 and 8.32 (2s, 1H), 7.57-7.35 (m, 1H), 7.31-7.15 (m, 1H), 7.10 (td, J= 7.9, 1.1 Hz, 1H), 5.37-5.11 (m, 2H), 4.92-4.18 (m, 3H), 3.83-3.16 (m, 2H), 2.28 (dd, J= 12.7, 8.7 Hz, 1H), 1.88-1.61 (m, 1H), 0.76 - 0.29 (m, 4H). ¹⁹F NMR (282 MHz, DMSO-J₆) δ 121.60; MS (ES+): 458.1 (M+l); (ES-): 456.1 (M-l), 492.1 (M+Cl).

Scheme 121

244

Préparation of (2S,4R)-l-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4methylpyrrolidine-2-carboxamide (121d)

Step-1: Préparation of (2S,4R)-tert-butyl 2-((3-chloro-2-fluorobenzyl)carbamoyl)-45 methylpyrrolidine-l-carboxylate (121b)

Compound 121b was prepared according to the procedure reported in step-1 of scheme-5, from (2S,4R)-l-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic acid (121a) (0.4 g, 1.745 mmol; CAS # 364750-80-1) using (3-chloro-2-fhiorophenyl)methanamine (109b) (0.658 mL, 5.23 mmol; CAS # 72235-55-3), HATU (995 mg, 2.62 mmol), DIPEA (0.912 mL, 5.23 mmol) 10 in DMF (5 mL) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with EtOAc/MeOH (ratio 9:1) in Hexanes from 0-100%] (2S,4R)-tert-butyl 2-((3-chloro-2-fluorobenzyl)carbamoyl)-4methylpyrrolidine-l-carboxylate (121b) as a white solid (0.55 g, 85% yield); MS (ES+): 393.1 (M+Na).

Step-2: Préparation of (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-methylpyrrolidine-2carboxamide (121c)

Compound 121c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-tert-butyl 2-((3-chloro-2-fluorobenzyl)carbamoyl)-4-methylpyrrolidine-1 carboxylate (121b) (0.55 g, 1.483 mmol) using TFA (1.143 mL, 14.83 mmol) in DCM (6 mL) and stirring ovemight at room température. This gave after workup and purification by reverse phase column chromatography [Cl8 (30 g), eluting with ACN in water (containing 0.1% HCl) from 0 to 100%] (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-methylpynOlidine-2-

245 carboxamide (121c) (0.34 g, 85% yield); *H NMR (300 MHz, DMSO-d₆) δ 9.92 (s, 1H), 9.15 (t, J=5.7Hz, 1H), 8.56 (s, 1H), 7.52 (ddd, J= 8.9, 7.2, 1.8 Hz, 1H), 7.39-7.29 (m, 1H), 7.22 (td,J=7.9, 1.1 Hz, 1H), 4.42 (d,J=5.7Hz, 2H), 4.32 (s, 1H), 3.47 - 3.36 (m, 1H), 2.87 -2.64 (m, 1H), 2.29 (h, .7=7.2 Hz, 1H), 2.12 - 1.99 (m, 1H), 1.99- 1.79 (m, 1H), 1.02 (d, J = 6.7 Hz, 3H); ^I9F NMR (282 MHz, DMSO-î/₆) δ -121.13; MS (ES+): 271.1 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-methylpyrrolidine-2-carboxamide (121d)

Compound 12Id was prepared according to the procedure reported in scheme-119, from (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-methylpyrrolidine-2-carboxamide (121c) (70.1 mg, 0.259 mmol) in DMF (5 mL) using 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (50 mg, 0.259 mmol) HOBT (17.49 mg, 0.129 mmol), EDC (99 mg, 0.518 mmol), DIPEA (0.180 mL, 1.035 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 to 100 %] followed by purification using reverse phase column chromatography [Cl8 column (15 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(6-amino9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-methylpynOlidine-2-carboxamide (121d) (11 mg, 9% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 8.89 and 8.52 (2t, 5.9 Hz, 1H), 8.35 and 8.33 (2s, 1H), 8.25 and 8.22 (2s, 1H), 7.53 - 7.33 (m,

1H), 7.21 (td, .7=7.2, 6.5, 1.9 Hz, 1H), 7.09 (td,J=7.9, 1.0 Hz, 1H), 5.20 (q, J= 17.0 Hz, 2H), 4.79 - 4.16 (m, 3H), 3.92 (t, J= 8.4 Hz, 1H), 3.73 - 3.56 (m, 1H), 3.22 (t, J= 9.3 Hz, 1H), 2.04- 1.91 (m, 1H), 1.86- 1.68 (m, 1H), 1.07 and 1.01 (2d, J=6.4 Hz, 3H). ¹⁹FNMR (282 MHz, DMSO-cZô) δ -121.70; MS (ES+): 446.1 (M+l), 468.1(M+Na); (ES-): 444.1 (ΜΙ), 480.1 (M+Cl).

Scheme 122

Préparation of (2S,4R)-1 -(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoro-4-methylpyrrolidine-2-carboxamide (122c)

246

Step-1: Préparation of (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoro-4-methylpyrrolidine-2carboxamide (122b)

Compound 122b was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-/ert-butyl 2-((3-chloro-2-fhiorobenzyl)carbamoyl)-4-fIuoro-4-methylpyrrolidine-1 carboxylate (122a) [(700 mg, 1.800 mmol); prepared according to the procedure reported by Altmann, Eva; et al., PCT Int. Appl. (2012), WO 2012093101 Al 20120712] using TFA (0.693 mL, 9.0 mmol) in DCM (7 mL) and stirring ovemight at room température. This gave after workup and purification by flash column chromatography [Cl8 (30 g), eluting with ACN in water (containing 0.1% HCl) from 0 to 100%] (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoro4-methylpyrrolidine-2-carboxamide (122b) (449 mg, 86% yield); 'H NMR (300 MHz, DMSOJ₆) δ 9.23 (t, J= 5.7 Hz, 1H), 7.53 (ddd, J = 8.0, 7.2, 1.8 Hz, 1H), 7.35 (ddd, J= 7.8, 6.7, 1.8 Hz, 1H), 7.23 (td, J =1.9, 1.1 Hz, 1H), 4.53 - 4.32 (m, 3H), 3.60-3.22 (m, 3H), 2.66 (td, J= 14.3, 7.5 Hz, 1H), 2.20- 1.92 (m, 1H), 1.54 (d, 7= 21.3 Hz, 3H); MS (ES+): 289.1 (M+l). Step-2: Préparation of (2S,4R)-l-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (122c)

Compound 122c was prepared according to the procedure reported in scheme-119, from (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (122b) (75 mg, 0.259 mmol) in DMF (5 mL) using 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (50 mg, 0.259 mmol) HOBT (17.49 mg, 0.129 mmol), EDC (99 mg, 0.518 mmol), DIPEA (0.180 mL, 1.035 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 to 100 %] followed by purification using reverse phase column chromatography [Cl8 column (15 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(6-amino-9Hpurin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (122c) (11 mg, 9% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMS0-7>) δ 9.06 and 8.67 (2t, 7=5.9 Hz, 1H), 8.37 (s, 1H), 8.31 (s, 1H), 7.57-7.37 (m, 1H), 7.25-7.13 (m, 1H), 7.07 (td, J = 7.9, 1.1 Hz, 1H), 5.45 - 5.03 (m, 2H), 4.92 - 3.57 (m, 5H), 2.44 (m, 1H), 2.14 1.82 (m, 1H), 1.55 (d, J= 21.1, 3H). ¹⁹F NMR (282 MHz, DMSO-7₆) δ -121.65, -139.64; MS (ES+): 464.1 (M+l), 486.1(M+Na); (ES-): 462.1 (M-l), 498.1 (M+Cl).

Scheme 123

247

Préparation of (2S,4R)-1 -(2-(6-((2-aminoethyl)amino)-9H-purin-9-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (123g)

Step-1: Préparation of ZerZ-butyl 2-(6-chloro-9H-purin-9-yl)acetate (123b)

Compound 123b was prepared according to the procedure reported in step-1 of scheme-1, from 6-chloro-9H-purine (123a) (8 g, 51.8 mmol; CAS # 87-42-3) in acetonitrile (400 mL) using ZerZ-butyl 2-bromoacetate (11.46 mL, 78 mmol), K2CO3 (14.31 g, 104 mmol) and heating at 65 °C for 48 h. This gave after workup and purification by flash column chromatography [silica gel (120 g), eluting with EtOAc in Hexane from 0-100%] ZerZ-butyl ΙΙΟ (6-chloro-9H-purin-9-yl)acetate (123b) (8.89 g, 64% yield) as an off-white solid; ’H NMR (300 MHz, DMSO-cfo) δ 8.81 (s, 1H), 8.68 (s, 1H), 5.16 (s, 2H), 1.42 (s, 9H); MS (ES+): 269.10 (M+l).

Step-2: Préparation of ZerZ-butyl 2-(6-((2-((ZerZ-butoxycarbonyl)amino)ethyl)amino)-9Hpurin-9-yl)acetate (123d)

248

A mixture of ZerZ-butyl 2-(6-chloro-9H-purin-9-yl)acetate (123b) (0.699 g, 2.6 mmol) and ZerZ-butyl 2-aminoethylcarbamate (123c) (0.961 g, 6.0 mmol; CAS # 57260-73-8) in EtOH (20 mL) was refluxed for 6 h. The solvent was evaporated under reduced pressure, and the product was purifïed by flash column chromatography [silica gel (40 g), eluting with DMA80 in DCM from 0 to 100 %] yielding ZerZ-butyl 2-(6-((2-((ZerZbutoxycarbonyl)amino)ethyl)amino)-9H-purin-9-yl)acetate (123d) (1.02 g, 100 % yield); *H NMR (300 MHz, DMSO-ri₆) δ 8.20 (s, 1H), 8.11 (s, 1H), 7.75 (s, 1H), 6.93 (s, 1H), 4.95 (s, 2H), 3.52 (s, 2H), 3.17 (q, J= 6.1 Hz, 2H), 1.39 (d, J= 16.4 Hz, 18H); MS (ES+): 393.20 (M+l).

Step-3: Préparation of 2-(6-((2-((ZerZ-butoxycarbonyl)amino)ethyl)amino)-9H-purin-9yl)acetic acid (123e)

To a stirred solution of ZerZ-butyl 2-(6-((2-((tert-butoxycarbonyl)amino)ethyl)amino)-9Hpurin-9-yl)acetate (123d) (1.02 g, 2.60 mmol) in THF (3.00 mL) and MeOH (3.00 mL) was added 2N sodium hydroxide (1.5 mL, 3.00 mmol) and stirred at room température for 45 min. This gave after workup 2-(6-(2-(ZerZ-butoxycarbonylamino)ethylamino)-9H-purin-9-yl)acetic acid (123e) (735mg, 84% yield); MS (ES+): 337.20 (M+l).

Step-4: Préparation of ZerZ-butyl (2-((9-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)4-fluoropyrrolidin-l-yl)-2-oxoethyl)-9H-purin-6-yl)amino)ethyl)carbamate (123f)

Compound 123f was prepared according to the procedure reported in step-3 of scheme-1, from 2-(6-((2-((ZerZ-butoxycarbonyl)amino)ethyl)amino)-9H-purin-9-yl)acetic acid (123e) (100 mg, 0.297 mmol) in DMF (2 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (7a) (82 mg, 0.297 mmol), HATU (170 mg, 0.446 mmol), DIPEA (0.207 mL, 1.189 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0-100%] ZerZ-butyl (2-((9-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-9H-purin-6-yl)amino)ethyl)carbamate (123f) (102 mg, 58% yield) HCl sait as a white solid; MS (ES+): 593.2 (M+l).

Step-5: Préparation of (2S,4R)-l-(2-(6-((2-aminoethyl)amino)-9H-purin-9-yl)acetyl)-N-(3chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (123g)

Compound 123g was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl (2-((9-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-9H-purin-6-yl)amino)ethyl)carbamate (1231) (lOOmg,

249

0.169 mmol) in DCM (2.5 mL) using TFA (0.260 mL, 3.37 mmol) and stirring at RT for 70 min. This gave after workup and purification by reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-1(2-(6-((2-aminoethyl)amino)-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (123g) (65 mg, 78% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-ri₆) δ 9.26 and 8.82 (2t, J= 5.9 Hz, 1H, D₂O exchangeable), 8.38 (d, J= 11.2 Hz, 2H, 1H D₂O exchangeable), 8.20 (s, 3H, 2H D₂O exchangeable), 7.53 - 7.35 (m, 1H), 7.35 - 7.13 (m, 1H), 7.05 (t, J= 7.9 Hz, 1H), 5.67 - 5.10 (m, 3H), 4.95 - 3.94 (m, 5H), 3.87 (m, 2H), 3.13 (m, 2H), 2.62 - 2.53 (m, 1H), 2.18 - 1.80 (m, 1H).¹⁹F NMR (282 MHz, DMSO-î/ô) δ -121.70, -176.23; MS (ES+): 493.1 (M+l); (ES-): 491.2 (M-l), 527.1 (M+Cl). Analysis calculated for C₂iH₂3ClF₂N₈O₂.1.875HC1.2H₂O: C, 42.23; H, 4.87; Cl, 17.06; N, 18.76; Found: C, 42.28; H, 4.81; Cl, 16.88; N, 18.54.

124a

Scheme 124

HATU, D1PEA

Préparation of (lR,3S,4S)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)2-azabicyclo[2.2. l]heptane-3-carboxamide (124b)

Compound 124b was prepared according to the procedure reported in step-3 of scheme-1, from (lR,3S,4S)-N-(3-chloro-2-fluorobenzyl)-2-azabicyclo[2.2.1]heptane-3-carboxamide (124a) (143 mg, 0.505 mmol; prepared according to the procedure reported by McDonald, Andrew; Qian, Shawn in PCT Int. Appl. (2018), WO 2018229543 A2 20181220) in DMF (1.5 mL) using 2-(6-amino-9H-purin-9-yl)acetic acid (67a) (65 mg, 0.337 mmol), HATU (192 mg, 0.505 mmol), DIPEA (0.293 mL, 1.683 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,4S)-2-(2-(6-amino-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-2azabicyclo[2.2.1]heptane-3-carboxamide (124b) (73 mg, 47% yield) HCl sait as a white

250 solid; 'H NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 8.98 (t, J= 5.8 Hz) and 8.56 (t, J= 5.9 Hz) (2t, 1H, D2O exchangeable), 8.49 (s) and 8.47 (s) (2s, 1H), 8.43 (s) and 8.35 (s), (2s, 1H), 7.51 - 7.03 (m, 3H), 5.46 (d, J= 17.0 Hz, 1H), 5.10 (d, J= 16.9 Hz, 1H), 4.79-4.16 (m, 3H), 3.84 (s, 1H), 2.58 (s, 1H), 1.99 (d, 7= 9.8 Hz, 1H), 1.88-1.21 (m, 5H);

¹⁹F NMR (282 MHz, DMSO-7₆) δ -121.38, -121.65. MS (ES+): 458.1 (M+l); (ES-): 456.7 (M-l); Analysis calculated for C21H21CIFN7O2.HCI.2.25H2O: C, 47.16; H, 4.99; Cl, 13.26; N, 18.33; Found: C, 47.34; H,4.76; Cl, 13.09; N, 18.16.

Scheme 125

Préparation of (2S,4R)-1 -(2-(4-amino-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (125d)

Step-1: Préparation of Zert-butyl 2-(4-amino-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetate (125b)

Compound 125b was prepared according to the procedure reported in step-1 of scheme-1, 15 from 5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-4-amine (125a) (0.5 g, 3.29 mmol; CAS #

1080467-52-2) in DMF (15 mL) using te/7-butyl 2-bromoacetate (0.510 mL, 3.45 mmol), CS2CO3 (1.285 g, 3.94 mmol) and stirring at RT for 1.5 h. This gave after workup tert-butyl 2-(4-amino-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (125b) (0.75 g, 86% yield) as an off-white solid; Ή NMR (300 MHz, DMSO-7₆) δ 8.05 (s, 1H), 7.13 (d, J= 2.3 Hz, 1H),

251

6.99 (s, 2H), 4.81 (s, 2H), 1.41 (s, 9H); ¹⁹FNMR(282 MHz, DMSO-7₆) δ-168.96; MS (ES+): 267.10 (M+l).

Step-2: Préparation of 2-(4-amino-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (125c)

Compound 125c was prepared according to the procedure reported in step-4 of scheme-17, from iert-butyl 2-(4-amino-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (125b) (720 mg, 2.70 mmol) in THF/MeOH (10 mL; ratio 1:1) using 2N sodium hydroxide (1.487 mL, 2.97 mmol) and stirring the reaction mixture for 1.5 h at RT. This gave after workup 2-(4amino-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (125c) (515 mg, 91% yield) as a 10 pale purple solid; Ή NMR (300 MHz, DMSO-7₆) δ 8.05 (s, 1H), 7.14 (d, J= 2.3 Hz, 1H), 6.99 (s, 2H), 4.83 (s, 2H); MS (ES+): 211.1 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropynOlidine-2-carboxamide (125d)

Compound 125d was prepared according to the procedure reported in step-3 of scheme-1, 75 from 2-(4-amino-5-fIuoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (125c) (50 mg, 0.238 mmol) in DMF (2 mL) using (2S,4R)-N-(3-chloro-2-fIuorobenzyl)-4-fluoropyrrolidine-2carboxamide (7a) HCl sait (74 mg, 0.238 mmol), HATU (113 mg, 0.297 mmol), DIPEA (0.166 mL, 0.952 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 20 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-1-(2-(4amino-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (125d) (59 mg, 53% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/₆) δ 9.19 (t, J= 5.7 Hz) and 8.75 (t, J= 5.9 Hz) (2t, 1H, D₂O exchangeable), 8.87 (s, 2H, D₂O exchangeable), 8.35 (s, 1H), 7.48 - 7.33 (m, 2H), 7.29 7.21 (m, 1H), 7.06 (td, J= 7.9, 1.1 Hz, 1H), 5.59 - 5.36 (m, 1H), 5.29 (d, 7 = 17.0 Hz, 1H), 5.12 (d, 7= 16.9 Hz, 1H), 4.48-3.76 (m, 5H), 2.50 - 2.38 (m, 1H), 2.22 - 1.90 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-7₆) δ -121.33, -121.64, -165.54, -176.28; MS (ES+): 467.0 (M+l); Calculated for C₂oHi₈C1F3N₆0₂.(H₂0).(HC1): C, 46.08; H, 4.06; Cl, 13.60; N, 16.12; Found:

C, 45.98; H, 3.98; Cl, 13.26; N, 15.77.

Scheme 126

252

HO_s

B'OH

Pd(PPh₃)CI₂, K₂CO₃

126b ^Boc

^Boc 126c ^Boc 126d

114b

Préparation of (2S,4R)-1 -(2-(4-amino-5-( 1 H-indol-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropynOlidine-2-carboxamide (126f)

Step-1: Préparation of tert-butyl 3-(4-amino-7-(2-(tert-butoxy)-2-oxoethyl)-7H-pyrrolo[2,35 d]pyrimidin-5-yl)-lH-indole-l-carboxylate (126c)

Compound 126c was prepared according to the procedure reported in step-1 of scheme-59, from /erZ-butyl 2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (114b) (290 mg, 0.886 mmol) in dioxane (8 mL) using (l-(Zert-butoxycarbonyl)-lH-indol-3-yl)boronic acid (126b) (231 mg, 0.886 mmol), bis(triphenylphosphine)palladium(II) chloride (93 mg, 10 0.133 mmol) a solution of potassium carbonate (368 mg, 2.66 mmol) in water (1 mL) and heating at 100 °C for 4 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0-50%] tert-butyl 3(4-ammo-7-(2-(tert-butoxy)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-lH-indole-lcarboxylate (126c) (165 mg, 40% yield) as a yellow solid; MS (ES+): 464.2 (M+l).

Step-2: Préparation of 2-(4-amino-5-(l-(tert-butoxycarbonyl)-lH-indol-3-yl)-7H-pynOlo[2,3d]pyrimidin-7-yl)acetic acid (126d) ’

Compound 126d was prepared according to the procedure reported in step-4 of scheme-17, from tert-butyl 3-(4-amino-7-(2-(tert-butoxy)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)IH-indole-l-carboxylate (126c) (160 mg, 0.345 mmol) in THF/MeOH (1.26 mL; ratio 1:1)

253 using 2N sodium hydroxide (0.190 mL, 0.380 mmol) and stirring for 1.5 h at RT. This gave after workup 2-(4-amino-5-(l-(tert-butoxycarbonyl)-lH-indol-3-yl)-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetic acid (126d) (97 mg, 69% yield) as a pale yellow solid; MS (ES+): 408.1 (M+l).

Step-3: Préparation of ierZ-butyl 3-(4-amino-7-(2-((2S,4R)-2-((3-chloro-2fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidin-5yl)-1 H-indole-1 -carboxylate (126e)

Compound 126e was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-5-(l-(teri-butoxycarbonyl)-lH-indol-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetic acid (126d) (60 mg, 0.147 mmol) in DMF (2 mL) using HCl sait of (2S,4R)-N-(3chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (7a) (45.8 mg, 0.147 mmol), HATU (70 mg, 0.184 mmol), DIPEA (0.103 mL, 0.589 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] teri-butyl 3-(4-amino-7-(2-((2S,4R)-2-((3chloro-2-fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l-yl)-2-oxoethyl)-7H-pyrrolo[2,3d]pyrimidin-5-yl)-lH-indole- 1-carboxylate (126e) (40 mg, 41% yield) as a white solid; MS (ES+): 664.2 (M+l).

Step-4: Préparation of (2S,4R)-l-(2-(4-amino-5-(lH-indol-3-yl)-7H-pyrrolo[2,3-d]pyrimidin7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropynOlidine-2-carboxamide (126f)

Compound 126f was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 3-(4-amino-7-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-lH-indole-lcarboxylate (126e) (39 mg, 0.059 mmol) in DCM (1 mL) using TFA (0.090 mL, 1.175 mmol) and stirring at RT for 4 h. This gave after workup and purification by reverse phase column chromatography [Cl8 column (50 g) eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-5-(lH-indol-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (126f) (15 mg, 45% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-Jô) (a mixture of two rotamers) δ 11.45 (d, J= 2.6 Hz, 1H), 9.08 (t, J= 5.7 Hz) and 8.64 (t, 5.9 Hz) (2t, 1H), 8.36 (d,

4.2 Hz, 1H), 7.56 - 7.30 (m, 6H, 2H D₂O exchangeable), 7.30 - 7.23 (m, 1H), 7.23 - 7.08 (m, 2H), 7.02 (q, J= 7.2 Hz, 1H), 6.92 (t, J= 7.9 Hz, 1H), 5.55 - 5.02 (m, 3H), 4.41 - 3.74 (m,

254

5H), 2.78 - 2.37 (m, 1H), 2.18-1.84 (m, 1H); ¹⁹F NMR (282 MHz, DMSO) δ -121.29, 121.68, -176.38; MS (ES+): 564.2 (M+l).

Scheme 127

Préparation of methyl l-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylate (127d)

Step-1: Préparation of methyl l-(2-(terr-butoxy)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5carboxylate (127b)

Compound 127b was prepared according to the procedure reported in step-1 of scheme-1, 10 from methyl lH-pyrrolo[2,3-b]pyridine-5-carboxylate (127a) (0.5 g, 2.84 mmol; CAS # 849067-96-5) in DMF (15 mL) using Zert-butyl 2-bromoacetate (0.419 mL, 2.84 mmol), CS2CO3 (1.110 g, 3.41 mmol) and stirring at RT for 1.5 h. This gave after workup methyl 1(2-(teri-butoxy)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylate (127b) (746 mg, 91% yield) as an off-white solid; Ή NMR (300 MHz, DMSO-îZô) δ 8.82 (d, J= 2.0 Hz, 1H), 8.58 15 (d, J= 2.0 Hz, 1H), 7.68 (d, J= 3.6 Hz, 1H), 6.68 (d, J= 3.6 Hz, 1H), 5.08 (s, 2H), 3.89 (s,

3H), 1.41 (s, 9H); MS (ES+): 235.00 (M+l).

Step-2: Préparation of 2-(5-(methoxycarbonyl)-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (127c)

255

Compound 127c was prepared according to the procedure reported in step-2 of scheme-1, from methyl l-(2-(iert-butoxy)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylate (127b) (400 mg, 1.378 mmol) in DCM (8 mL) using TFA (1.582 mL, 20.67 mmol) and stirring at RT for 16 h. This gave after workup 2-(5-(methoxycarbonyl)-lH-pyrrolo[2,3-b]pyridin-lyl)acetic acid (127c) (470 mg, 98% yield) TFA sait as a yellow solid; ‘H NMR (300 MHz, DMSO-î/ô) δ 8.82 (d, J= 2.0 Hz, 1H), 8.58 (d, J= 2.0 Hz, 1H), 7.69 (d, J= 3.6 Hz, 1H), 6.67 (d, J= 3.6 Hz, 1H), 5.10 (s, 2H), 3.89 (s, 3H); MS (ES+): 235.10 (M+l).

Step-3: Préparation of methyl l-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylate (127d)

Compound 127d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(5-(methoxycarbonyl)-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (127c) (75 mg, 0.215 mmol) in DMF (2 mL) using (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) HCl sait (68.6 mg, 0.215 mmol), HATU (123 mg, 0.323 mmol), DIPEA (0.188 mL, 1.077 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] methyl l-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylate (127d) (95mg, 89% yield) as a white solid; Ή NMR (300 MHz, DMSO-tZ₆) δ 10.78 (s, 1H, D₂O exchangeable), 8.82 (d, J= 2.0 Hz, 1H), 8.56 (d, J= 2.0 Hz, 1H), 8.03 (d, J= 8.2 Hz, 1H), 7.72 (t, J= 8.0 Hz, 1H), 7.64 (d, J= 3.6 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 6.66 (d, J= 3.6 Hz, 1H), 5.57 (d, J= 17.1 Hz, 1H), 5.31 (d, J= 17.0 Hz, 1H), 4.44 (dd, J= 9.0, 5.5 Hz, 1H), 3.89 (s, 3H), 3.79 (ddd, J= ΊΑ, 5.5, 2.3 Hz, 1H), 2.39 - 2.11 (m, 2H), 1.89 (tt, J= 6.5, 3.7 Hz, 1H), 1.03 (dt, J= 8.8, 5.4 Hz, 1H), 0.70 (td, J= 5.2, 2.3 Hz, 1H); MS (ES+): 497.9 (M+l), 519.9 (M+Na); (ES-): 496.0 (M-l); Analysis calculated for C₂₂H₂oBrN504.1.75H₂0: C, 49.87; H, 4.47; N, 13.22; Found: C, 49.80; H, 4.09; N, 13.01.

Scheme 128

Préparation of l-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylic acid (128a)

Compound 128a was prepared according to the procedure reported in step-4 of scheme-17, from methyl l-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylate(127d) (67mg, 0.134 mmol) in THF (0.8 mL), acetonitrile (0.4 mL) and water (2 mL) using IN aqueous lithium hydroxide monohydrate (0.403 mL, 0.403 mmol) and stirring at RT for 16 h.

This gave after workup and purification by reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] l-(2((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylic acid (128a) (6 mg, 9% yield) HCl sait as a white solid; ’H NMR (300 MHz, Methanol-iL) δ 8.96 - 8.84 (m, 2H), 8.02 (dd, J = 12.4, 8.6

Hz, 1H), 7.62 (m, 2H), 7.25 (dt, J = 7.5, 3.4 Hz, 1H), 6.86 (dt, J = 5.9, 3.1 Hz, 1H), 5.76 5.36 (m, 2H), 4.53 (d, J = 8.6 Hz, 1H), 3.77 (m, 1H), 2.43 (q, J = 6.0, 4.2 Hz, 1H), 2.13 1.80 (m, 2H), 1.27 - 1.11 (m, 1H), 1.03 - 0.76 (m, 1H); MS (ES+): 483.9 (M+l), 505.9 (M+Na); (ES-): 481.9 (M-l).

Scheme 129

Préparation of (1R,3S,5R)-2-(2-(5-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (129f)

Step-1: Préparation of teri-butyl 2-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (129b)

Compound 129b was prepared according to the procedure reported in step-1 of scheme-1, from 5-bromo-7H-pyrrolo[2,3-d]pyrimidine (129a) (1 g, 5.05 mmol; CAS # 175791-49-8) m DMF (25 mL) using rert-butyl 2-bromoacetate (0.784 mL, 5.30 mmol), CS2CO3 (1.974 g, 6.06 mmol) and stirring at RT for 1.5 h. This gave after workup rert-butyl 2-(5-bromo-7Hpyrrolo[2,3-d]pyrimidin-7-yl)acetate (129b) (1.54 g, 98% yield) as a pale-yellow solid; 'H ₁₀ NMR (300 MHz, DMSO-*) δ 8.96 (s, 1H), 8.90 (s, 1H), 7.89 (s, 1H), 5.06 (s, 2H), 1.42 (s,

9H); MS (ES+): 312.00 (M+l).\

Step-2: Préparation of teri-butyl 2-(5-((teri-butoxycarbonyl)a_mino)-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetate (129c)

Compound 129c was prepared according to the procedure reported in step-3 of scheme-17, ,5 from fôrt-butyl 2-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (129b) (500 mg, 1.602 mmol) in toluene (30 mL) using XPhos (153 mg, 0.320 mmol), t-butyl carbamate (563 mg, 4.81 mmol), Pda(dba)₃ (147 mg, 0.160 mmol), césium carbonate (522 mg, 1.602 mmol) and heating at 95 °C for 16 h. This gave after work up and purification tert-butyl 2-(5-((tertbutoxycarbonyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (129c) (29 mg, 5% yield);

258

Ή NMR (300 MHz, DMSO-d₆) δ 9.89 (s, 1H), 9.30 (s, 1H), 8.88 (s, 1H), 7.80 (s, 1H), 5.05 (s, 2H), 1.52 (s, 9H), 1.42 (s, 9H); MS (ES+): 349.20 (M+l).

Step-3: Préparation of 2-(5-((tert-butoxycarbonyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetic acid (129d)

Compound 129d was prepared according to the procedure reported in step-4 of scheme-17, from tert-butyl 2-(5-((tert-butoxycarbonyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (129c) (80 mg, 0.23 mmol) in THF (2 mL) and MeOH (2 mL) using 1 N solution of aqueous lithium hydroxide hydrate (1.148 mL, 1.148 mmol) and stirring at RT for 15 h. This gave after workup 2-(5-((tert-butoxycarbonyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (129d) (62 mg, 92% yield); *H NMR (300 MHz, DMSO-d₆) δ 9.89 (s, 1H), 9.30 (s, 1H), 8.87 (s, 1H), 7.80 (s, 1H), 5.06 (s, 2H), 1.52 (s, 9H); MS (ES+): 293.10 (M+l).

Step-4: Préparation of tert-butyl (7-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)carbamate (129e)

Compound 129e was prepared according to the procedure reported in step-3 of scheme-1, 75 from 2-(5-((tert-butoxycarbonyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (129d) (50 mg, 0.171 mmol) in DMF (1 mL) using TFA sait of (lR,3S,5R)-N-(6-bromopyridin-2yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (74.5 mg, 0.188 mmol), HATU (98 mg, 0.257 mmol), DIPEA (0.149 mL, 0.855 mmol) and stirring at RT for 4 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), DMA-80 in 20 DCM from 0 -100%] tert-butyl (7-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)carbamate (129e) (85 mg, 89% yield) as a white solid; ^!H NMR (300 MHz, DMSO-dô) δ 10.77 (s, 1H), 9.77 (s, 1H), 9.19 (s, 1H), 8.73 (s, 1H), 8.04 (d, J = 8.2 Hz, 1H), 7.72 (t, J = 7.9 Hz, 1H), 7.64 (s, 1H), 7.33 (d, J = 7.7 Hz, 1H), 5.47 (d, J = 17.1 Hz, 1H), 5.20 (d, J = 17.0 Hz, 1H), 4.47 - 4.33 (m, 25 1H), 3.75 (s, 1H), 2.34 - 2.11 (m, 2H), 1.89 (d, J = 12.1 Hz, 1H), 1.51 (s, 9H), 1.03 (t, J = 9.3

Hz, 1H), 0.72 (s, 1H); MS (ES+): 556.20 (M+l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(5-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (129f)

Compound 129f was prepared according to the procedure reported in step-2 of scheme-1, 30 from tert-butyl (7-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)carbamate (129e) (80 mg, 0.144 mmol) in DCM (0.8 mL) using TFA (0.222 mL, 2.88 mmol) and stirring at RT

259 for 16 h. This gave after workup (lR,3S,5R)-2-(2-(5-amino-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (129f) (59 mg, 90 % yield) HCl sait as a yellow solid; ^XH NMR (300 MHz, DMSO-dô) δ 10.80 (s, 1H, D₂O exchangeable), 9.34 (s, 1H), 9.04 (s, 1H), 8.03 (d, J = 8.1 Hz, 1H), 7.77 (s, 1H), 7.72 (t, J =

8.0 Hz, 1H), 7.34 (d, J = 7.6 Hz, 1H), 5.65 (d, J = 17.1 Hz, 1H), 5.34 (d, J = 17.0 Hz, 1H),

4.45 (dd, J = 9.0, 5.4 Hz, 1H), 3.78 (m, 1H), 2.26 (m, 2H), 1.90 (p, J = 6.7 Hz, 1H), 1.04 (m, 1H), 0.73 (m, 1H). MS (ES+): 456.1 (M+l); (ES-): 454.1 (M-l); Analysis calculated for Ci9Hi8BrN₇O2 2.25H2O.2.25HCl: C, 39.42; H, 4.31; Cl, 13.78; N, 16.94; Found: C, 39.7; H, 4.28; Cl, 13.43; N, 16.71.

Scheme 130

Préparation of (2S,4R)-l-(2-(4-amino-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (130e)

Step-1: Préparation of ZerZ-butyl 2-(4-chloro-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-715 yl)acetate (130b)

Compound 130b was prepared according to the procedure reported in step-1 of scheme-1, from 4-chloro-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (130a) (2 g, 11.20 mmol; CAS # 24391-41-1) in DMF (50 mL) using sodium hydride (0.538 g, 13.44 mmol), ZerZ-butyl 2bromoacetate (1.986 mL, 13.44 mmol) and stirring at RT for 19 h. This gave after workup

ZerZ-butyl 2-(4-chloro-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (130b) (2.005 g, 61%

260 yield) as a white solid; Ή NMR (300 MHz, DMSO-J6) δ 8.86 (s, 1H), 8.71 (s, 1H), 5.17 (s, 2H), 1.41 (s, 9H); MS (ES+): 293.05 (M+l).

Step-2: Préparation of 2-(4-chloro-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (130c)

Compound 130c was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-chloro-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (130b) (1.98 g, 6.76 mmol) in DCM (60 mL) using TFA (5.21 mL, 67.6 mmol) and stirring at RT for 20 h. This gave after workup 2-(4-chloro-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (130c) (2.38 g) as an off-white solid which was used as such for next step.

Step-3: Préparation of (2S,4R)-N-(3-chloro-2-fluorobenzyl)-l-(2-(4-chloro-5-cyano-7Hpyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-4-fluoropyrrolidine-2-carboxamide (130d)

Compound 130d was prepared according to the procedure reported in step-3 of scheme-1, from of 2-(4-chloro-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (130c) (516 mg, 2.182 mmol) in DMF (30 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-415 fluoropyrrolidine-2-carboxamide (7a) (472 mg, 1.718 mmol), HATU (1307 mg, 3.44 mmol), DIPEA (1110 mg, 8.59 mmol) and stirring at RT for 21 h. This gave after workup and purification by flash column chromatography [silica gel, eluting with MeOH:EtOAc (1:9) in hexanes] (2S,4R)-N-(3-chloro-2-fluorobenzyl)-l-(2-(4-chloro-5-cyano-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetyl)-4-fluoropyrrolidine-2-carboxamide (130d) (155 mg, 18%) as a yellow solid; ²Η NMR (300 MHz, DMSO-dô) (a mixture of two rotamers) δ 9.03 (t) & 8.67 8.57 (m) (2H), 8.82 & 8.80 (2s, 1H),7.54 - 7.32 (m, 1H), 7.26 - 7.12 (m, 1H), 7.09 - 6.97 (m, 1H), 5.68 - 5.18 (m, 3H), 4.97 - 3.71 (m, 5H), 2.61 - 2.35 (m, 1H), 2.21 - 1.92 (m, 1H); ^I9F NMR (282 MHz, DMSO-dô) (a mixture of two rotamers) δ -121.32 & -121.57 (1F), -176.13 &-176.41 (1F) MS (ES+): 493.10 & 495.00 (M+l).

Step-4: Préparation of (2S,4R)-l-(2-(4-amino-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (130e)

To a solution of (2S,4R)-N-(3-chloro-2-fluorobenzyl)-l-(2-(4-chloro-5-cyano-7Hpyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-4-fluoropyrrolidine-2-carboxamide (130d) (100 mg, 0.203 mmol) in dioxane (4 mL) was added ammonium hydroxide (3.84 mL, 56.8 mmol) and 30 heated at 85 °C in a sealed tube for 14 h. The reaction mixture was concentrated to dryness and purified by flash column chromatography [silica gel, eluting with MeOH in DCM from

261

0-5%] to afford (2S,4R)-l-(2-(4-amino-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (130e) (37 mg, 39% yield) as a white solid. Ή NMR (300 MHz, DMSO-dô) (a mixture of two rotamers) δ 9.02 & 8.60 (2t, J = 5.9 Hz, 1H, D₂O exchangeable), 8.18 & 8.17 (2s, 1H), 8.10 & 8.07 (2s, 1H), 7.53 - 7.33 (m, 1H), 7.27 - 7.13 (m, 1H), 7.10 - 7.01 (m, 1H), 6.86 (s, 2H, D₂O exchangeable), 5.60 -

5.36 (m, 1H), 5.32 - 5.02 (m, 2H), 4.93 - 3.58 (m, 5H), 2.62-2.34 (m, 1H), 2.31 - 1.85 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-ife) (a mixture of two rotamers) δ -121.36 & -121.64, 176.12 & -176.39; MS (ES+): 474.10 (M+l); MS (ES-): 472.60 (M-l).

Scheme 131

Préparation of (2S,4R)-l-(2-(4-amino-lH-pyrrolo[2,3-b]pyridin-l-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (131e)

Step-1: Préparation of tert-butyl 2-(4-chloro-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (131b)

Compound 131b was prepared according to the procedure reported in step-1 of scheme-1, 15 from 4-chloro-lH-pyrrolo[2,3-b]pyridine (131a) (1 g, 6.55 mmol; CAS # 55052-28-3) in

DMF (25 mL) using sodium hydride (0.315 g, 7.86 mmol), tert-butyl 2-bromoacetate (1.162 mL, 7.86 mmol) and stirring at RT for 14 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with EtOAc in hexane from 0 to 85 %] tertbutyl 2-(4-chloro-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (131b) (1.45 g, 83% yield) as a yellow solid; ’H NMR (300 MHz, DMSO-d₆) δ 8.21 (d, J = 5.2 Hz, 1H), 7.66 (d, J = 3.6 Hz,

262

1H), 7.26 (d, J = 5.2 Hz, 1H), 6.56 (d, J = 3.6 Hz, 1H), 5.05 (s, 2H), 1.40 (s, 9H); MS (ES+): 267.10 & 269.10 (M+l).

Step-2: Préparation of tert-butyl 2-(4-((teri-butoxycarbonyl)amino)-lH-pyrrolo[2,3b]pyridin-l-yl)acetate (131c)

Compound 131c was prepared according to the procedure reported in step-3 of scheme-17, from (tert-butyl 2-(4-chloro-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (131b) (1 g, 3.75 mmol) in toluene (30 mL) using XPhos (0.179 g, 0.375 mmol), t-butyl carbamate (0.659 g, 5.62 mmol), Pd₂(dba)3 (0.172 g, 0.187 mmol), césium carbonate (1.222 g, 3.75 mmol) and heating at 90 °C for 19 h. This gave after work up and purification by flash column chromatography

[silica gel, EtOAc in Hexane from 0 -14% then 33%] ie/7-butyl 2-(4-((tertbutoxycarbonyl)amino)-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (131c) (1.145 g, 88% yield) as a yellow gum; 'H NMR (300 MHz, DMSO-d₆) δ 9.72 (s, 1H), 8.05 (d, J = 5.5 Hz, 1H), 7.61 (d,

J = 5.5 Hz, 1H), 7.32 (d, J = 3.6 Hz, 1H), 6.90 (d, J = 3.6 Hz, 1H), 4.94 (s, 2H), 1.53 (s, 9H), 15 1.40 (s, 9H); MS (ES+): 348.20 (M+l).

Step-3: Préparation of 2-(4-amino-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (131d)

Compound 131d was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-((ieri-butoxycarbonyl)amino)-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (131c) (1 g, 2.88 mmol) in DCM (30 mL) using TFA (1.711 mL, 23.03 mmol) and stirring at

RT for 18 h. This gave after work up 2-(4-amino-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (131d) (1.62 g) as a yellow gum; MS (ES+): 192.10 (M+l).

Step-4: Préparation of (2S,4R)-l-(2-(4-amino-lH-pyrrolo[2,3-b]pyridin-l-yl)acetyl)-N-(3chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (13 le)

Compound 131e was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (131d) (83 mg, 0.432 mmol) in DMF (10 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2carboxamide (7a) (148 mg, 0.540 mmol), HATU (329 mg, 0.864 mmol), DIPEA (0.376 mL, 2.160 mmol) and stirring at RT for 18 h. This gave after workup and purification using reverse phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-lH-pyrrolo[2,3-b]pyridin-lyl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (131e) (23 mg, 12% yield) HCl sait as a light yellow solid; ⁴H NMR (300 MHz, DMSO-dô) (a mixture of two

263 rotamers) δ 14.17 - 13.83 (m, 1H), 9.23 - 8.79 (m, 1H), 8.16 (s, 2H), 7.97 - 7.74 (m, 1H), 7.57 - 7.35 (m, 1H), 7.32 - 7.08 (m, 2H), 6.96 - 6.79 (m, 2H), 6.51 - 6.45 (m, 1H), 5.62 5.24 (m, 3H), 4.99 - 3.54 (m, 5H), 2.73 - 2.42 (m, 1H), 2.37 - 1.86 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-ί/δ) (a mixture of two rotamers) δ -121.29 & -121.79, -175.81 & -175.96; MS (ES+): 448.10 (M+l); MS (ES-): 446.70 (M-l).

Scheme 132

Préparation of (2S,4R)-l-(2-(4-amino-2-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (132d)

Step-1: Préparation of tert-butyl 2-(4-amino-2-methyl-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetate (132b)

Compound 132b was prepared according to the procedure reported in step-1 of scheme-1, from 2-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (132a) (500 mg, 3.37 mmol) in DMF (15 ml.) using tert-butyl 2-bromoacetate (0.498 mL, 3.37 mmol), CS2CO3 (1.319 g, 4.05 mmol) and stirring at RT for 18 h. This gave after workup and purification by flash column chromatography [silica gel, eluting with MeOH in DCM from 0-5%, then 10% ] tert-butyl 2(4-amino-2-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (132b) (700 mg, 79% yield) as off-white solid; Ή NMR (300 MHz, DMSO-d₆) δ 7.01 (d, J = 3.5 Hz, 1H), 6.87 (s, 2H), 6.46 (d, J = 3.5 Hz, 1H), 4.80 (s, 2H), 2.33 (s, 3H), 1.41 (s, 9H); MS (ES+): 263.10 (M+l).

264

Step-2: Préparation of 2-(4-amino-2-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (132c)

Compound 132c was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-2-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (132b) (670 mg, 2.55 mmol) using TFA. This gave after workup 2-(4-amino-2-methyl-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetic acid (132c) (875 mg) off white solid; MS (ES+): 207.05 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-2-methyl-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (132d)

Compound 132d was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-2-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (132c) (89 mg, 0.432 mmol) in DMF (10 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2carboxamide (7a) (148 mg, 0.540 mmol), HATU (329 mg, 0.864 mmol), DIPEA (0.376 mL, 2.160 mmol) and stirring at RT for 12 h. This gave after workup and purification using reverse phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-2-methyl-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (132d) (34 mg, 17% yield) HCl sait as a white solid; ^XH NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 13.94 (s, 1H, D₂O exchangeable), 9.26 (s, 1H, D2O exchangeable), 9.15 - 8.61 (m, 1H, D₂O exchangeable), 8.07 (s, 1H, D₂O exchangeable),

7.51 - 7.35 (m, 1H), 7.33 - 7.20 (m, 2H), 7.16 & 7.06 (2t, J = 7.9 Hz, 1H), 6.93 - 6.86 (m,

1H), 5.64 - 4.94 (m, 3H), 4.92 - 3.69 (m, 5H), 2.77 - 2.25 (m, 4H), 2.27 - 1.89 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-ifc) (a mixture of two rotamers) δ -121.33 & -121.66 (1F), -176.06 & -176.34 (1F); MS (ES+): 463.10 & 465.10 (M+l).

Scheme 133

265

Préparation of (2S,4R)-l-(2-(4-amino-5-(lH-indol-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (133b)

Compound 133b was prepared according to the procedure reported in step-1 of scheme-59, from (2S,4R)-l-(2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (116b) (200 mg, 0.379 mmol), 1HIndole-2-boronic acid pinacol ester (133a) (92 mg, 0.379 mmol), Pd(PPh3)2C12 (26.6 mg, 0.038 mmol) in dioxane (5 mL) using a 3.3 M aqueous K2CO3 (0.345 mL, 1.137 mmol) and heating at 100 °C for 16 h under argon. This gave after workup and purification by flash column chromatography (SiO₂,12 g, eluting with 0-5% MeOH in DCM) followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-5-(lH-indol-2yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (133b) (37 mg, 17 % yield) HCl sait as a pale-orange solid;

’H NMR (300 MHz, DMSO-dô) (mixture of two rotamers) δ 11.65 - 11.50 (m, 1H, D₂O exchangeable), 8.69 (t, J = 5.9 Hz, 1H), 8.45 (t, J = 2.3 Hz, 1H), 7.89 (s, 2H, D₂O exchangeable), 7.66 (dd, J = 3.8,1.2 Hz, 1H), 7.59 (d, J = 7.8 Hz, 1H), 7.52 - 7.40 (m, 1H), 7.40 - 7.29 (m, 1H), 7.29 - 7.20 (m, 1H), 7.19 - 7.10 (m, 1H), 7.04 (dt, J = 12.1, 7.7 Hz, 2H), 6.59 (d, J = 2.0 Hz, 1H), 5.67 - 5.10 (m, 3H), 4.41 (m, 2H), 4.31 - 4.09 (m, 1H), 3.90 (m, 1H),

2.22 - 1.93 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-de) δ -121.30, -121.63, -176.11, -176.32;

MS (ES+) 564/566 (M+l), (ES-) 560/562 (M-l); Analysis calculated for C₂8H24C1F₂N7O2*HC1«2.75H₂O: C, 51.74; H, 4.73; Cl, 10.91; N, 15.08; Found: C, 51.74; H, 4.56; Cl, 10.98; N, 15.07.

Scheme 134

Préparation of (2S,4R)-l-(2-(4-amino-5-(lH-indol-7-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (134b)

Compound 134b was prepared according to the procedure reported in step-1 of scheme-59, from (2S,4R)-l-(2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (116b) (200 mg, 0.379 mmol), 1HIndole-7-boronic acid pinacol ester (134a) (92 mg, 0.379 mmol), Pd(PPh3)₂Cl₂ (26.6 mg, 0.038 mmol) in dioxane (5 mL) using a 3.3 M aqueous K2CO3 (0.345 mL, 1.137 mmol) and heating at 100 °C for 16 h under argon. This gave after workup and purification by flash column chromatography (SiO₂,12 g, eluting with 0-5% MeOH in DCM) followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-5-(lH-indol-7yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (134b) (47 mg, 22 % yield) HCl sait as a white solid; ⁴H

NMR (300 MHz, DMSO-ifo) (mixture of two rotamers) δ 11.03 (q, J = 4.1, 3.2 Hz, 1H, D₂O exchangeable), 8.68 (t, J = 5.9 Hz, 1H), 8.45 (d, J = 3.7 Hz, 1H), 7.64 (dt, J = 7.7,1.7 Hz, 1H), 7.56 (d, J = 6.4 Hz, 1H), 7.50 - 7.30 (m, 2H, D₂O exchangeable), 7.30 - 7.20 (m, 1H), 7.15 (t, J = 7.5 Hz, 1H), 7.11 - 7.06 (m, 1H), 7.01 - 6.92 (m, 1H), 6.57 (dd, J = 3.1,1.7 Hz, 1H), 5.62 - 5.13 (m, 3H), 4.52 - 4.31 (m, 2H), 4.29 - 4.11 (m, 2H), 3.88 (m, 1H), 2.21 -1.92 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-d₆) δ -121.29, -121.67, -176.13, -176.29; MS (ES+):

564/566 (M+l), (ES-): 560/562 (M-l); Analysis calculated for C28H24ClF2N7O2H.25HCh2.5H2O: C, 51.37; H, 4.66; Cl, 12.19; N, 14.98; Found: C, 51.69; H, 4.49; Cl, 11.79; N, 14.99.

Scheme 135

267

116b

Préparation of (2S,4R)-l-(2-(4-amino-5-phenyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (135a)

Compound 135a was prepared according to the procedure reported in step-1 of scheme-59, 5 from (2S,4R)-l-(2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (116b) (200 mg, 0.379 mmol), phenylboronic acid (46.2 mg, 0.379 mmol), Pd(PPh₃)2C12 (26.6 mg, 0.038 mmol) in dioxane (5 mL) using a 3.3 M aqueous K2CO3 (0.345 mL, 1.137 mmol) and heating at 100 °C for 16 h under argon. This gave after workup and purification by flash column chromatography (S1O2, 10 12 g, eluting with 0-5% MeOH in DCM) followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%](2S,4R)-l-(2-(4-amino-5-phenyl-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (135a) (53 mg, 27 % yield) HCl sait as a white solid after lyophilization. ¹H NMR (300 MHz, DMSO-ifo) (mixture of two rotamers) δ 8.69 (t, J = 6.0 Hz, 1H), 8.45 (d, J = 4.3 Hz, 1H), 7.97 - 7.34 (m, 9H), 7.22 (m, 1H), 7.00 (m, 1H), 5.58 - 5.16 (m, 3H), 4.51 - 4.09 (m, 5H), 4.02 - 3.77 (m, 1H), 2.21 -1.92 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-d₆) δ -121.29, -121.64, -176.12, 176.32; MS (ES+): 525/527 (M+l), (ES-): 521/523 (M-l); Analysis calculated for C26H23C1F₂N₆O2‘HC1«2H2O: C, 52.27; H, 4.72; Cl, 11.87; N, 14.07; Found: C, 52.01; H,

4.69; Cl, 11.63; N, 13.99.

Scheme 136

268

Pd(PPh₃)₂CI₂ K₂CO₃

Préparation of (2S,4R)-l-(2-(4-amino-5-(2-chlorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (136a)

Compound 136a was prepared according to the procedure reported in step-1 of scheme-59, from (2S,4R)-l-(2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (116b) (200 mg, 0.379 mmol), 2chlorophenylboronic acid (59.3 mg, 0.379 mmol), Pd(PPh3)zC12 (26.6 mg, 0.038 mmol) in dioxane (5 mL) using a 3.3 M aqueous K2CO3 (0.345 mL, 1.137 mmol) and heating at 100 °C for 16 h under argon. This gave after workup and purification by flash column chromatography (SiO₂,12 g, eluting with 0-5% MeOH in DCM) followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with 0-60% ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-5-(2-chlorophenyl)7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2carboxamide (136a) (43 mg, 20 % yield) HCl sait as a white solid after lyophilization. ^JH

NMR (300 MHz, DMSO-Ts) (mixture of two rotamers) δ 8.70 (t, J = 5.9 Hz, 1H), 8.43 (d, J = 4.3 Hz, 1H), 7.83 - 7.61 (m, 2H), 7.54 - 7.35 (m, 6H), 7.30 - 7.23 (m, 1H), 7.02 (t, J = 7.9 Hz, 1H), 5.61 - 5.17 (m, 3H), 4.51 - 4.09 (m, 5H), 4.02 - 3.77 (m, 1H), 2.21 -1.91 (m, 1H). ¹⁹F ' NMR (282 MHz, DMSO-d₆) δ -121.28, -121.65, -176.10, -176.29; MS (ES+) 559/561 (M+l), (ES-) 555/557 (M-l); Analysis calculated for C26H22C12F2N6O2’HC1*2H2O: C, 49.42;

H, 4.31; Cl, 16.83; N, 13.30; Found: C, 49.20; H, 4.24; Cl, 16.72; N, 13.24.

Scheme 137

Préparation of (2S,4R)-l-(2-(4-amino-5-(lH-indol-6-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (137b)

Compound 137b was prepared according to the procedure reported in step-1 of scheme-59, from (2S,4R)-l-(2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro2-fhiorobenzyl)-4-fluoropyrrolidine-2-carboxamide (116b) (200 mg, 0.379 mmol), indole-6boronic acid pinacol ester (137a) (92 mg, 0.379 mmol), Pd(PPh3)2Ch (26.6 mg, 0.038 mmol) in dioxane (5 mL) using a 3.3 M aqueous K2CO3 (0.345 mL, 1.137 mmol) and heating at 100 °C for 16 h under argon. This gave after workup and purification by flash column chromatography (SiO₂,12 g, eluting with 0-5% MeOH in DCM) followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with 0-60% ACN in water (containing 0.1% HCl) from 0-100%](2S,4R)-l-(2-(4-amino-5-(lH-indol-6-yl)-7Hpyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2carboxamide (137b) (30 mg, 14 % yield) HCl as a white solid; ³Η NMR (300 MHz, DMSO15 dô) (mixture of two rotamers) δ 11.30 (d, J = 2.3 Hz, 1H, D2O exchangeable), 8.67 (t, J = 5.9 Hz, 1H), 8.43 (d, J = 4.0 Hz, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.51 - 7.41 (m, 3H), 7.36 (m, 1H), 7.28 - 7.18 (m, 1H), 7.11 (m, 1H), 7.05 - 6.96 (m, 1H), 6.51 (t, J = 2.5 Hz, 1H), 5.78 - 5.13 (m, 2H), 4.93 - 4.32 (m, 2H), 4.32 - 4.06 (m, 1H), 3.89 (m, 1H), 2.22 -1.90 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-rfc) δ -121.28, -121.64, -176.09, -176.30; MS (ES+): 564/566 (M+l), (ES-): 560/562 (M-l); Analysis calculated for C2sH24ClF2N7O2.HCl.2H2O: C, 52.10;

H, 4.68; Cl, 10.98; N, 15.19; Found: C, 51.79; H, 4.31; Cl, 11.23; N, 15.29.

Scheme 138

270

Préparation of (2S,4R)-l-(2-(4-aminoquinazolin-8-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (138d)

Step-1: Préparation of ZerZ-butyl 2-(4-aminoquinazolin-8-yl)acetate (138b)

Compound 138b was prepared according to the procedure reported in step-1 of scheme-2, from 8-bromoquinazolin-4-amine (138a) (1.00 g, 4.46 mmol; CAS # 1260657-19-9) using zinc (1.752 g, 26.8 mmol), TMSC1 (0.283 mL, 2.232 mmol) in THF (5 mL), Pd₂(dba)₃ (0.409 g, 0.446 mmol), XPhos (0.426 g, 0.893 mmol) in THF (10 mL). This gave after work up and purification using flash column chromatography [silica gel (24 g), eluting with EtOAc in hexane from 0-50%] ZerZ-butyl 2-(4-aminoquinazolin-8-yl)acetate (138b) (97 mg, 8.4%) as a yellow solid; Ή NMR (300 MHz, DMSO-Je) δ 8.39 (s, 1H), 8.12 (dd, J = 8.4, 1.4 Hz, 1H), 7.90 - 7.68 (m, 2H), 7.65 (dd, J = 7.0,1.2 Hz, 1H), 7.42 (dd, J = 8.3, 7.1 Hz, 1H), 3.94 (s, 2H), 1.39 (s, 9H); MS (ES+): 260 (M+l).

Step-2: Préparation of 2-(4-aminoquinazolin-8-yl)acetic acid (138c)

Compound 138c was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-aminoquinazolin-8-yl)acetate (138b) (95 mg, 0.366 mmol) using TFA (2.103 mL, 5.50 mmol) in DCM (5 mL) and stirring at RT for 16 h. This gave after work up 2-(2-(4-aminoquinazolin-8-yl)acetic acid (138c) (106 mg, 91%) TFA sait as an orange solid; Ή NMR (300 MHz, DMSO-d₆) δ 9.88 - 9.57 (m, 2H), 8.76 (s, 1H), 8.36 (dd, J = 8.4,1.2 Hz,

271

1H), 7.97 (dd, J = 7.4,1.2 Hz, 1H), 7.73 (t, J = 7.8 Hz, 1H), 4.07 (s, 2H). ¹⁹F NMR (282

MHz, DMSO-Jô) δ -74.37; LC-MS: t = 0.56 min; MS (ES+): 204 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-aminoquinazolin-8-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (138d)

Compound 138d was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-aminoquinazolin-8-yl)acetic acid (138c) (93 mg, 0.293 mmol) in DMF (5 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (7a) (114 mg, 0.293 mmol), HATU (134 mg, 0.352 mmol), DIPEA (0.490 mL, 2.81 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography (SiO₂, 12 g, eluting with 0-3% MeOH in DCM) followed by reverse phase column chromatography [C-18 column, 100 g, eluting with 0.1% aqueous HCl in H₂O and MeCN from 0-100%] (2S,4R)-l-(2-(4-aminoquinazolin-8-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (138d) (22 mg, 16 % yield) HCl sait as a pale-yellow solid; ⁴Η NMR (300 MHz, DMSO-îL) (mixture of two rotamers) δ 9.80 (s, 2H, D2O exchangeable), 8.82 (q, J = 5.6, 4.1 Hz, 1H, D2O exchangeable), 8.52 (s, 1H), 8.39 (d, J = 8.3 Hz, 1H), 7.91 (d, J = 7.3 Hz, 1H), 7.72 (t, J = 7.8 Hz, 1H), 7.40 (td, J = 7.6,1.6 Hz, 1H), 7.30 - 7.15 (m, 1H), 6.92 (t, J = 8.2 Hz, 1H), 5.65 - 5.19 (m, 1H), 4.50 - 3.90 (m, 9H), 2.25 -1.95 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-d6) δ -121.15, -121.69, -175.82; MS (ES+): 460/462 (M+l), (ES-): 457/459 (M-l).

Scheme 139

H N

H N

Pd(PPh₃)₂CI₂ K₂CO₃

139b

116b

Préparation of (2S,4R)-l-(2-(4-amino-5-(lH-indol-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (139b)

272

Compound 139b was prepared according to the procedure reported in step-1 of scheme-59, from (2S,4R)-l-(2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (116b) (200 mg, 0.379 mmol), indole-5boronic acid pinacol ester (139a) (92 mg, 0.379 mmol), Pd(PPh3)2C12 (26.6 mg, 0.038 mmol) in dioxane (5 mL) using a 3.3 M aqueous K2CO3 (0.345 mL, 1.137 mmol) and heating at 100 °C for 16 h under argon. This gave after workup and purification by flash column chromatography (S1O2,12 g, eluting with 0-3% MeOH in DCM) followed by purification using reverse phase column chromatography [Cl 8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%](2S,4R)-l-(2-(4-amino-5-(lH-indol-5-yl)-7H10 pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2carboxamide (139b) (73 mg, 34 % yield) HCl sait as a white solid; ¹H NMR (300 MHz, DMSO-dô) (a mixture of two rotamers) δ 11.31 (d, J = 2.6 Hz, 1H, D2O exchangeable), 8.67 (t, J = 6.0 Hz, 1H), 8.42 (d, J = 4.0 Hz, 1H), 7.63 (d, J = 1.6 Hz, 1H), 7.54 (d, J = 8.4 Hz, 1H), 7.44 (m, 3H), 7.37 (m, 1H), 7.29 - 7.22 (m, 1H), 7.19 (m, 2H), 7.02 (t, J = 7.9 Hz, 1H), 6.51 (d, J = 2.6 Hz, 1H), 5.64 - 5.10 (m, 4H), 4.41 (m, 3H), 4.20 (m, 2H), 4.03 - 3.78 (m, 1H), 2.22

-1.93 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-de) δ -121.29, -121.64, -176.11, -176.30; MS (ES+): 564/566 (M+l), (ES-): 560/562 (M-l); Analysis calculated for

C28H24CIF2N7O2.HCI.2.25H2O: C, 52.47; H, 4.64; Cl, 11.06; N, 15.30 Found: C, 52.27; H, 4.40; Cl, 11.42; N, 15.41.

Préparation of 4-amino-7-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (140a)

Compound 140a was prepared according to the procedure reported in step-1 of scheme-60, 25 from (2S,4R)-l-(2-(4-amino-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro21436

273 ,

2-fhiorobenzyl)-4-fluoropyrrolidine-2-carboxamide (130e) (150 mg, 0.317 mmol) in éthanol (8 mL) using conc. NH4OH (3 mL) and hydrogen peroxide (0.112 mL, 1.266 mmol) and stirring at RT for 5 h. This gave after workup and purification by reverse phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 5 0-100%] 4-amino-7-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4-fluoropyrrolidinl-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxâmide (140a) (134 mg, 86% yield)

HCl sait as a white solid; Ή NMR (300 MHz, DMSO-deÆhO) (a mixture of two rotamers) δ 8.81 - 8.65 (m, 1H), 8.38 (d, J = 4.6 Hz, 1H), 8.15 (s, 1H), 7.42 (q, J = 10.2, 9.0 Hz, 1H), 7.21 (q, J = 8.7, 8.1 Hz, 1H), 7.07 (t, J = 7.9 Hz, 1H), 5.64 - 5.11 (m, 3H), 4.93 - 3.87 (m, 10 5H), 2.51 (s, 1H), 2.23 - 1.85 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-d₆) (a mixture of two rotamers) δ -121.33 & -121.61, -176.20 &-176.51; MS (ES+): 492.10, 494.10 (M+l);

Analysis calculated for C21H20CIF2N7O3.HQ.2H2O: C, 44.69; H, 4.46; N, 17.37; Cl, 12.56;

Found: C, 44.49; H, 4.44; N, 17.34; Cl, 12.42.

Scheme 141

Préparation of (2S,4R)-l-(2-(4-amino-5-(aminomethyl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (141a)

To a cooled solution of (2S,4R)-l-(2-(4-amino-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (130e) (300 mg, 20 0.633 mmol) in methanol (20 mL) was added nickel(II) chloride hexahydrate (37.6 mg, 0.158 mmol) followed by sodium borohydride (144 mg, 3.80 mmol) in several portions over a period of 10 min and stirred at RT for 1 h. To this mixture was added Nl-(2aminoethyl)ethane-l,2-diamine (0.137 mL, 1.266 mmol) and stirred at RT for 0.5 h and concentrated under vacuum. The residue was treated with ethyl acetate (120 mL), washed 25 with water (60 mL) and the aqueous phase was extracted again with ethyl acetate (60 mL).

274

The combined organics were washed with brine (75 mL), dried, filtered and concentrated in vacuum. The obtained residue was purified by reverse phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] to afford (2S,4R)-l-(2-(4-amino-5-(aminomethyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(35 chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (141a) (48 mg, 16% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-rfô) (a mixture of two rotamers) δ 9.18 8.53 (m, 3H, D2O exchangeable), 8.39 - 8.36 (m, 1H), 8.23 (s, 3H, D2O exchangeable), 7.52 &7.51 (2s, 1H), 7.49 - 7.05 (m, 3H), 5.65 - 5.07 (m, 3H), 4.99 - 3.72 (m, 7H), 2.61 - 2.40 (m, 1H), 2.21 - 1.88 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î/ô) (a mixture of two rotamers) δ

-121.32 & -121.61, -176.11 & -176.32; MS (ES+): 478.10 & 480.10 (M+l); Analysis calculated for C21H22CIF2N7O2.I.85 HC1.1.75 H₂O: C, 43.72; H, 4.78; N, 17.00; Cl, 17.52; Found: C, 43.89; H, 4.62; N, 16.93; Cl, 17.84.

i

Scheme 142

Préparation of (2S,4R)-l-(2-(4-amino-6-methyl-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetyl)-N(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (142d)

Step-1: Préparation of ZerZ-butyl 2-(4-amino-6-methyl-lH-pyrazolo[3,4-d]pyrimidin-lyl)acetate (142b)

275

Compound 142b was prepared according to the procedure reported in step-1 of scheme-1, from 6-methyl-lH-pyrazolo[3,4-d]pyrimidin-4-amine (142a) (500 mg, 3.35 mmol; CAS # 5326-80-7) in DMF (15 mL) using tert-butyl 2-bromoacetate (0.495 mL, 3.35 mmol), CS2CO3 (1.311 g, 4.02 mmol) and stirring at RT for 18 h. This gave after workup and 5 purification by flash column chromatography [silica gel; eluting with MeOH in DCM from 05%] tert-butyl 2-(4-amino-6-methyl-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetate (142b) (360 mg, 41% yield) as a yellow solid; NMR (300 MHz, DMSO-Jô) δ 8.03 (s, 1H), 7.62 (s, 2H), 4.97 (s, 2H), 2.37 (s, 3H), 1.40 (s, 9H); MS (ES+): 264.1 (M+l).

Step-2: Préparation of 2-(4-amino-6-methyl-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetic acid 10 (142c)

Compound 142c was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-6-methyl-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetate (142b) (335 mg, 1.272 mmol) using TFA in DCM (20 mL). This gave after workup 2-(4-amino-6-methyllH-pyrazolo[3,4-d]pyrimidin-l-yl)acetic acid (142c) (715 mg) and used as such for the next 75 step; MS (ES+): 208.10 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-6-methyl-lH-pyrazolo[3,4-d]pyrimidin-lyl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (142d)

Compound 142d was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-methyl-lH-pyrazolo[3,4-d]pyrimidin-l-yl)acetic acid (142c) (0.432 20 mmol) in DMF (10 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2carboxamide (7a) (0.540 mmol), ELATU (0.329 g, 0.864 mmol), DIPEA (0.376 mL, 2.160 mmol) and stirring at RT for 20 h. This gave after workup and purification using reverse phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-6-methyl-lH-pyrazolo[3,4-d]pyrimidin-l25 yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (142d) (92 mg, 46% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/₆) (a mixture of two rotamers) δ 10.00 (s, 1H, D2O exchangeable), 9.09 (t, J = 5.9 Hz) & 8.73 - 8.57 (m) (2H, D₂O exchangeable), 8.45 & 8.44 (2s, 1H), 7.52 - 7.41 (m, 1H), 7.40 - 7.19 (m, 1H), 7.16 & 7.06 (2t, J = 7.9 Hz, 1H), 5.62 - 5.17 (m, 3H), 4.96 - 3.73 (m, 5H), 2.54 & 2.52 (2s, 3H), 30 2.60 - 2.40 (m, 1H), 2.19 -1.90 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î/₆) (a mixture of two rotamers) δ -121.30 & -121.65, -176.29 & -176.51; MS (ES+): 464.10 & 466.10 (M+l);

276

Analysis calculated for C20H20CIF2N7O2.HCI.2.25H2O: C, 44.41; H, 4.75; Cl, 13.11; N, 18.13; Found: C, 44.52; H, 4.47; Cl, 12.80; N, 17.95.

Scheme 143

Préparation of (2S,4R)-l-(2-(4-amino-5-(furan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (143d)

Step-1: Préparation of ZerZ-butyl 2-(4-amino-5-(furan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetate (143b)

Compound 143b was prepared according to the procedure reported in step-1 of scheme-59, 10 from ZerZ-butyl 2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (114b) (500 mg, 1.528 mmol) in dioxane (15 mL) using furan-3-ylboronic acid (143a) (171 mg, 1.528 mmol), bis(triphenylphosphine)palladium(II) chloride (161 mg, 0.229 mmol) a solution of potassium carbonate (634 mg, 4.58 mmol) in water (1.8 mL) and heating at 100 °C for 11 h. This gave after workup and purification by flash column chromatography [silica gel, eluting 15 with EtOAc in hexane from 0-67% followed by 10% methanol in EtOAc (1:1)] ZerZ-butyl 2(4-amino-5-(furan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (143b) (190 mg, 40% yield) as a yellow solid; ³H NMR (300 MHz, DMSO-ί/β) δ 8.11 (s, 1H), 7.83 (s, 1H), 7.81 — 7.78 (m, 1H), 7.27 (s, 1H), 6.69 (dd, J = 1.8, 0.9 Hz, 1H), 6.25 (s, 2H), 4.90 (s, 2H), 1.42 (s, 9H); MS (ES+): 315.10 (M+l).

277

Step-2: Préparation of 2-(4-amino-5-(furan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (143c)

Compound 143c was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-5-(furan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (143b) (175 mg, 0.557 mmol) using TFA in DCM (10 mL). This gave after workup 2-(4-amino-5(furan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (143c) which was used as such for the next step; MS (ES+): 259.05 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-5-(furan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fhiorobenzyl)-4-fluoropyrrolidine-2-carboxamide (143d)

Compound 143d was prepared according to the procedure reported in step-3 of scheme-1, 2(4-amino-5-(furan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (143c) (0.557 mmol) in DMF (15 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2carboxamide (7a) (0.668 mmol), ELATU (0.424 g, 1.114 mmol), DIPEA (0.485 mL, 2.79 mmol) and stirring at RT for 13 h. This gave after workup and purification using flash column chromatography [silica column, DCM in MeOH from 0-5 %] followed by purification using reverse phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-5-(furan-3-yl)-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (143d) (30 mg, 10% for two steps) HCl sait as a white βοϋά^Η NMR (300 MHz, DMSO-dô) (a mixture of two rotamers) δ 9.11 & 8.68 (2t, J = 5.9 Hz, 1H), 8.43 & 8.42 (2s, 1H), 7.91 — 7.89 (m, 1H), 7.88 - 7.83 (m, 1H), 7.51 &7.50 (2s, 1H), 7.49 - 6.97 (m, 3H), 6.72 - 6.69 (m, 1H), 5.63 - 5.08 (m, 3H), 4.93 - 3.70 (m, 5H), 2.67-2.30 (m, 1H), 2.23 - 1.89 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-de) (a mixture of two rotamers) δ -121.31 & -121.64, -176.12 & 176.34; MS (ES+): 515.10 & 517.10 (M+l); Analysis calculated for

C24H21CIF2N6O3.HCI.I.75H2O: C, 49.45; H, 4.41; N, 14.42; Cl, 12.16; Found: C, 49.31; H,

4.13; N, 14.34; Cl, 11.94.

Scheme 144

278

144a 144b 144c

F,

Cl ζ Cl

144f 144g

Préparation of 4-amino-l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxamide (144g)

Step-1: Préparation of tert-butyl 2-(4-chloro-5-cyano-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate 5 (144b)

Compound 144b was prepared according to the procedure reported in step-1 of scheme-88, from 4-chloro-lH-pyrrolo[2,3-b]pyridine-5-carbonitrile (144a) (950 mg, 5.35 mmol; CAS # 920966-02-5) in DMF (25 mL) using sodium hydride (0.257 g, 6.42 mmol), tert-butyl bromoacetate (0.949 mL, 6.42 mmol) and stirring at RT for 18 h. This gave after workup and 10 purification by flash column chromatography [silica gel, EtOAc in hexane from 0 -14%] tert-butyl 2-(4-chloro-5-cyano-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (144b) (970 mg, 62% yield) as a white solid; ^XH NMR (300 MHz, DMSO-ifc) δ 8.73 (s, 1H), 7.88 (d, J = 3.7 Hz, 1H), 6.78 (d, J = 3.6 Hz, 1H), 5.12 (s, 2H), 1.40 (s, 9H). MS (ES+): 292.10 & 294.10 (M+l).

279

Step-2: Préparation of ZerZ-butyl 2-(4-((ZerZ-butoxycarbonyl)amino)-5-cyano-lH-pyrrolo[2,3b]pyridin-l-yl)acetate (144c)

Compound 144c was prepared according to the procedure reported in step-3 of scheme-17, from ZerZ-butyl 2-(4-chloro-5-cyano-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (144b) (950 mg,

3.26 mmol) in toluene (30 mL) using XPhos (155 mg, 0.326 mmol), t-butyl carbamate (572 mg, 4.88 mmol), Pd₂(dba)s (149 mg, 0.163 mmol), césium carbonate (1061 mg, 3.26 mmol) and heating at 90 °C for 19 h. This gave after work up and purification by flash column chromatography [silica gel, EtOAc in hexane from 0 - 33%] ZerZ-butyl 2-(4-((tertbutoxycarbonyl)amino)-5-cyano-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (144c) (1.13 g, 93 % yield); Ή NMR (300 MHz, DMSO-d₆) δ 10.19 (s, 1H), 8.51 (s, 1H), 7.61 (d, J = 3.7 Hz, 1H), 6.68 (d, J = 3.6 Hz, 1H), 5.04 (s, 2H), 1.51 (s, 9H), 1.40 (s, 9H); MS (ES+): 373.20 (M+l).

Step-3: Préparation of 2-(4-((ZerZ-butoxycarbonyl)amino)-5-cyano-lH-pyrrolo[2,3-b]pyridinl-yl)acetic acid (144d)

Compound 144d was prepared according to the procedure reported in step-4 of scheme-17, from ZerZ-butyl 2-(4-((ZerZ-butoxycarbonyl)amino)-5-cyano-lH-pyrrolo[2,3-b]pyridin-lyl)acetate (144c) (1.1g, 2.95 mmol) in THF (5 mL) and MeOH (5 mL) using 2M sodium hydroxide (1.698 mL, 3.40 mmol) and stirring at RT for 25 h. This gave after workup 2-(4((ZerZ-butoxycarbonyl)amino)-5-cyano-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (144d) (782 mg, 84% yield) as a light yellow solid; MS (ES+): 317.10 (M+l).

Step-4: Préparation of ZerZ-butyl (l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-5-cyano-lH-pyrrolo[2,3-b]pyridin-4-yl)carbamate (144e)

Compound 144e was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-((ZerZ-butoxycarbonyl)amino)-5-cyano-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (144d) (600 mg, 1.897 mmol) in DMF (30 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)25 4-fluoropyrrolidine-2-carboxamide (7a) (625 mg, 2.276 mmol), HATU (1442 mg, 3.79 mmol), DIPEA (1.652 mL, 9.48 mmol) and stirring at RT for 13 h. This gave after workup and purification by flash column chromatography [silica gel, eluting with MeOH in DCM from 0 - 5%] ZerZ-butyl (l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-5-cyano-lH-pyrrolo[2,3-b]pyridin-4-yl)carbamate (144e) (855 mg, 79% yield) as a white solid; ^XH NMR (300 MHz, DMSO-de) (a mixture of two rotamers) δ 10.20 (s, 1H), 9.03 & 8.60 (2t, J = 5.8 Hz, 1H), 8.473 & 8.467 (2s, 1H), 7.56 21436

280

7.37 (m, 2H), 7.28 - 7.13 (m, 1H), 7.09 - 7.00 (m, 1H), 6.69 & 6.66 (2d, J = 3.6 Hz, 1H), 5.68 - 5.05 (m, 3H), 4.91 - 3.66 (m, 5H), 2.61 - 2.04 (m, 2H), 1.52 (s, 9H); ¹⁹F NMR (282 MHz, DMSO-J6) (a mixture of two rotamers) δ -121.32 & -121.71 (1F), -176.11 & -176.35 (1F); MS (ES+): 573.20 & 575.20 (M+l);

Step-5: Préparation of teri-butyl (5-carbamoyl-l-(2-((2S,4R)-2-((3-chloro-2fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridin-4yl)carbamate (144f)

Compound 144f was prepared according to the procedure reported in step-1 of scheme-60, from ieri-butyl (l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l10 yl)-2-oxoethyl)-5-cyano-lH-pyrrolo[2,3-b]pyridin-4-yl)carbamate (144e) (150 mg, 0.262 mmol) in éthanol (8 mL) using conc. NH4OH (3 mL), hydrogen peroxide (0.093 mL, 1.047 mmol) and stirring for 5 h at RT. This gave after workup and purification by flash column chromatography [silica gel, eluting with MeOH in DCM from 0 - 5%] ieri-butyl (5carbamoyl-l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l-yl)-215 oxoethyl)-lH-pyrrolo[2,3-b]pyridin-4-yl)carbamate (144f) (42 mg, 27% yield) as a white solid; Ή NMR (300 MHz, DMSO-rfô) (a mixture of two rotamers) δ11.15&11.14 (2s, 1H), 9.02 & 8.60 (2t, 1H), 8.51 & 8.50 (2s, 1H), 8.23 (s, 1H), 7.62 (s, 1H), 7.53 - 7.37 (m, 1H), 7.34 & 7.31 (2d, J = 3.6 .

Hz, 1H), 7.28 - 7.15 (m, 1H), 7.11 - 7.02 (m, 1H), 6.72 & 6.70 (2d, J = 3.8 Hz, 1H), 5.60 20 5.36 (m, 1H), 5.34 - 5.03 (m, 2H), 4.93 - 3.65 (m, 5H), 2.63 - 1.77 (m, 2H), 1.50 (s, 9H); ¹⁹F

NMR (282 MHz, DMSO-î/ô) (a mixture of two rotamers) δ -121.36 & -121.74 (1 F) , -176.10 & -176.38 (1 F); MS (ES+): 591.10 & 593.20 (M+l);

Step-6: Préparation of 4-amino-l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxamide (144g)

Compound 144g was prepared according to the procedure reported in step-2 of scheme-1, from ieri-butyl (5-carbamoyl-l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridin-4-yl)carbamate (144f) (40 mg, 0.068 mmol) in DCM (10 mL) using TFA (0.101 mL, 1.354 mmol) and stirring at RT for 25 h. This gave after workup and purification by reverse phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] 4-amino-l(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l-yl)-2-oxoethyl)lH-pyrrolo[2,3-b]pyridine-5-carboxamide (144g) (21 mg, 63% yield) HCl sait as a white

281 solid; ^]H NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 9.09 & 8.76 (2t, J = 6.1 Hz, 1H, D2O exchangeable), 8.47 & 8.42 (2s, 1H), 8.16 (s, 1H, D2O exchangeable), 7.59 (s, 1H, D₂O exchangeable), 7.53 - 7.33 (m, 1H), 7.30 - 6.85 (m, 4H), 5.62 - 5.56 & 5.45 - 5.39 (2m, 1H), 5.37 (s, 2H), 4.91 - 3.81 (m, 5H), 2.64 - 2.41 (m, 1H), 2.31 - 1.93 (m, 1H); ¹⁹F

NMR (282 MHz, DMSO-î/ô) (a mixture of two rotamers) δ -121.31 & -121.66 9, -175.78 & 175.96; MS (ES+): 491.10 & 493.10 (M+l).

Scheme 145

Préparation of (2S,4R)-N-(3-chloro-2-fluorobenzyl)-l-(2-(2,4-dioxo-3,4-dihydro-lH10 pyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-7(2H)-yl)acetyl)-4-fluoropyrrolidine-2-carboxamide (145a)

Compound 145a was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl (l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-lyl)-2-oxoethyl)-5-cyano-lH-pyrrolo[2,3-b]pyridin-4-yl)carbamate (144e) (100 mg, 0.175 mmol) in DCM (10 mL) using TFA (0.104 mL, 1.396 mmol) and stirring at RT for 22 h. This gave after workup and purification by reverse phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-N-(3chloro-2-fluorobenzyl)-l-(2-(2,4-dioxo-3,4-dihydro-lH-pyrrolo[3',2':5,6]pyrido[4,3d]pyrimidin-7(2H)-yl)acetyl)-4-fluoropyrrolidine-2-carboxamide (145a) (8 mg, 9% yield) as a white solid; Ή NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 11.84 (s, 1H, D₂O exchangeable), 11.35 (s, 1H, D₂O exchangeable), 9.08 - 8.99 & 8.62 - 8.56 (2t, 1H, D2O exchangeable), 8.64 (s, 1H) 7.52 — 6.92 (m, 5H), 5.60 — 5.11 (m, 3H), 4.95 — 3.69 (m, 5H), 2.78 - 1.84 (m, 2H); ¹⁹F NMR (282 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 121.32 &-121.71, -176.10 & -176.32; MS (ES+): 517.10 & 519.10 (M+l).

Scheme 146

282

130e

146a

Préparation of methyl 4-amino-7-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylate (146a)

To a solution of (2S,4R)-l-(2-(4-amino-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N5 (3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (190 mg, 0.401 mmol) (130e) in methanol (10 mL) was added conc. sulfuric acid (0.214 mL, 4.01 mmol) and refluxed for 43 h. The reaction mixture was cooled to RT diluted with ethyl acetate (100 mL) and 0.5 M K2CO3 (40 ml). The organic layer was washed with brine (50 mL), dried, filtered and concentrated in vacuum. The residue obtained was purified using reverse-phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] to afford methyl 4-amino-7-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylate HCl sait (146a) (1.3 mg, 0.64% yield) HCl sait as a white solid. ^XH NMR (300 MHz, DMSO-dô) (a mixture of two rotamers) δ 9.11 - 8.42 (m, 3H, D₂O exchangeable), 8.39 - 8.35 (m, 1H), 8.17 & 8.16 (2s, 1H), 7.57 - 6.99 (m, 3H), 5.65 - 5.08 (m, 3H), 4.88 - 3.91 (m, 5H), 3.88 (s, 3H),

2.62 - 2.36 (m, 1H), 2.24 - 1.87 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-d₆) (a mixture of two rotamers) δ -121.64, -176.14; MS (ES+): 507.10 & 509.10 (M+l).

Scheme 147

283

Préparation of methyl 4-amino-l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylate (147a) i

I

Compound 147a was prepared according to the procedure reported in scheme-146, from tert5 butyl (l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l-yl)-2oxoethyl)-5-cyano-lH-pyrrolo[2,3-b]pyridin-4-yl)carbamate (144e) (200 mg, 0.349 mmol) in methanol (10 mL) using conc. sulfuric acid (0.186 mL, 3.49 mmol) and refluxing for 14 h. This gave after work up and purification using reverse-phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] methyl 410 amino-l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l-yl)-2oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylate (147a) (2.3 mg, 1.3% yield) HCl sait as a white solid; ^:Η NMR (300 MHz, DMSO-de) (a mixture of two rotamers) δ 9.03 & 8.64 (2t, J = 5.8 Hz, 1H, D2O exchangeable), 8.52 & 8.44 (2s, 1H), 7.56 - 6.83 (m, 5H), 5.63 - 5.07 (m, 3H), 4.93 - 3.71 (m, 8H), 2.68 - 2.37 (m, 1H), 2.32 - 1.90 (m, 1H); ¹⁹F NMR (282 MHz,

DMSO-dô) (a mixture of two rotamers) δ -121.33 & -121.68, -175.91 &-176.15; MS (ES+): 506.10 & 508.15 (M+l).

Scheme 148

148d

Préparation of (2S,4R)-l-(2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(320 chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (148d)

284

Step-1: Préparation of teri-butyl 2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (148b)

Compound 148b was prepared according to the procedure reported in step-1 of scheme-1, from 5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (148a) (5 g, 19.23 mmol; CAS # 1636225 50-2) in DMF (20 mL) using tert-butyl 2-bromoacetate (3.75 g, 19.23 mmol), CS2CO3 (7.52 g, 23.07 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] ieri-butyl 2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (148b) (5 g, 70% yield) as a pale orange solid; Ή NMR (300 MHz, DMSO-d₆) δ 8.09 (s, 1H), 7.43 (s, 1H), 6.86 - 6.41 (m, 10 2H), 4.87 (s, 2H), 1.41 (s, 9H). MS (ES+): 375 (M+l).

Step-2: Préparation of 2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (148c)

Compound 148c was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (148b) (2.50 g, 6.68 mmol) using TFA (38.3 mL, 100 mmol; 20% TFA in DCM) and stirring at RT for 16 h. 15 This gave after workup 2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (148c) (2.91 g) TFA sait as a purple solid; ³H NMR (300 MHz, DMSO-î/ô) δ 8.32 (d, J = 1.2 Hz, 1H), 7.66 (d, J = 1.3 Hz, 1H), 4.97 (s, 2H). ¹⁹F NMR (282 MHz, DMSO-de) δ -74.24; MS (ES+): 319 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-720 yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (148d)

Compound 148d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (148c) (2.88 g, 6.67 mmol) in DCM (10 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (7a) (2.59 g, 6.67 mmol), HATU (3.04 g, 8.00 mmol), 25 DIPEA (5.82 mL, 33.3 mmol) and stirring at RT for 16 h. This gave after workup and purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (2S,4R)-l-(2-(4-amino-5-iodo-7Hpyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2carboxamide (148d) (55 mg, 0.096 mmol) HCl sait as a white solid; ^XH NMR (300 MHz, 30 DMSO-ifc) δ (a mixture of two rotamers) 9.08 (t, J = 5.9 Hz) and 8.67 (t, J = 5.9 Hz) (2t, 1H), 8.36 (d, J = 3.8 Hz, 1H), 8.01 (s, 2H, D₂O exchangeable), 7.61 (d, J = 5.6 Hz, 1H), 7.52 7.34 (m, 1H), 7.29 - 7.13 (m, 1H), 7.08 (t, J = 8.1 Hz, 1H), 5.64 - 5.36 (m, 1H), 5.29 (d, J =

285

17.1 Hz, 1H), 5.19 - 5.03 (m, 1H), 4.50 - 4.31 (m, 2H), 4.31 - 4.03 (m, 2H), 3.85 (ddd, J = 38.4,12.6, 3.1 Hz, 1H), 2.60 - 2.21 (m, 1H), 2.20 - 1.90 (m, 1H); ^{10 * * * * 15 * * * 19 20 * * * * 25}F NMR (282 MHz, DMSO) δ -121.32, -121.62, -176.13, -176.34; MS (ES+): 575.0 (M+l); (ES-): 573.0 (M-l); Analysis calculated for C20Hi8ClF2IN6O2.HCl.2H2O: C, 37.11; H, 3.58; Cl, 10.96; N, 12.98:

Found: C, 37.13; H, 3.20; Cl, 10.93; N, 13.12. ’

Scheme 149

116b 149b

Préparation of (2S,4R)-l-(2-(4-amino-5-(lH-pyrrol-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (149b)

Compound 149b was prepared according to the procedure reported in step-1 of scheme-62, from (2S,4R)-l-(2-(4-amino-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(3-chloro2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (116b) (200 mg, 0.379 mmol) in dioxane (5 mL) using 2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrrole (149a) (110 mg,

0.568 mmol; CAS # 476004-79-2), Pd(PPh₃)2Cl₂ (26.6 mg, 0.038 mmol), 3.3 M aqueous

K₂CO₃ (0.345 mL, 1.137 mmol) and heating at 100 °C for 16 h under argon. This gave after workup and purification using flash column chromatography [silica column (12 g), eluting with MeOH in DCM from 0-5 %] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-60%] (2S,4R)-l-(2-(4-amino-5-(lH-pyrrol-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-720 yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (149b) (27 mg,14 % yield) HCl sait as a white solid; ^!H NMR (300 MHz, DMSO-îZ₆) δ (a mixture of two rotamers)11.31 (d, J = 8.3 Hz, 1H, D₂O exchangeable), 9.08 (t), 8.65 (t, J = 5.8 Hz) (2t,lH),

8.39 (d, J = 3.6 Hz, 1H), 7.64 (s, 2H, D2O exchangeable), 7.50 - 7.34 (m, 2H), 7.29 - 7.13 (m, 1H), 7.02 (t, J = 7.9 Hz, 1H), 6.91 (p, J = 2.0 Hz, 1H), 6.22 (t, J = 2.2 Hz, 2H), 5.66 25 5.04 (m, 3H), 4.56 - 4.07 (m, 4H), 4.01 - 3.72 (m, 1H), 2.62 - 2.37 (m, 1H), 2.21 - 1.89 (m,

286

1H); ¹⁹F NMR (282 MHz, DMSO-de) δ -121.31, -121.64, -176.11, -176.31; MS (ES+) 514.0 (M+l), 512.0 (M-l); Analysis calculated for: CiÆbzClFzNîOz-HClXSOHzO: C, 48.41; H, 4.74; Cl, 11.91; N, 16.47; Found: C, 48.22; H, 4.49; Cl, 12.01; N, 16.36.

Scheme 150

Préparation of (2S,4R)-l-(2-(4-amino-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidme-2-carboxamide (150d)

Step-1: Préparation of tert-butyl 2-(4-amino-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetate (150b)

Compound 150b was prepared according to the procedure reported in step-1 of scheme-1, from 5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (150a) (0.5 g, 3.37 mmol; CAS # 150110-6) in DMF (20 mL) using tert-butyl 2-bromoacetate (0.658 g, 3.37 mmol), CS2CO3 (1.319 g, 4.05 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] tert-butyl

2-(4-amino-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (150b) (0.30 g, 34% yield) as a pale orange solid; ^H NMR (300 MHz, DMSO-t/e) δ 7.99 (s, 1H), 6.86 (d, J — 1.3 Hz, 1H), 6.55 (s, 2H), 4.78 (s, 2H), 2.34 (s, 3H), 1.41 (s, 9H).

Step-2: Préparation of 2-(4-amino-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (150c) .

287

Compound 150c was prepared according to the procedure reported in step-2 of scheme-1, from ieri-butyl 2-(4-amino-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (150b) (300 mg, 1.144 mmol) using TFA (9781 mg, 17.16 mmol; 20% TFA in DCM) and stirring at RT for 16 h. This gave after workup TFA sait of 2-(4-amino-5-methyl-7H-pyrrolo[2,35 d]pyrimidin-7-yl)acetic acid (150c) (470 mg) as a pale-orange solid; ¹H NMR (300 MHz, DMSO-Je) δ 8.68 - 8.38 (m, 2H), 8.35 (s, 1H), 7.26 (d, J = 1.3 Hz, 1H), 4.98 (s, 2H), 2.41 (d, J = 1.2 Hz, 3H).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (150d)

Compound 150d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (150c) (150 mg, 0.468 mmol) in DMF (5 mL) using TFA sait of (2S,4R)-N-(3-chloro-2fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (7a) (182 mg, 0.468 mmol), HATU (214 mg, 0.562 mmol), DIPEA (0.409 mL, 2.342 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (150d) (83 mg, 38% yield) as a white solid; ^]H NMR (300 MHz, DMSO-d₆) δ (a mixture of two rotamers) 9.15 (t, J = 5.8 Hz) and 8.71 (t, J = 5.9 Hz) (2s,1H), 8.47 (s, 2H, D₂O exchangeable), 8.34 (d, J = 3.2 Hz, 1H), 7.53 - 7.40 (m, 1H), 7.31 - 7.21 (m, 1H), 7.20 - 7.11 (m, 1H), 7.07 (t, J = 7.9 Hz, 1H), 5.59 - 5.33 (m, 1H), 5.26 (d, J = 17.0 Hz, 1H), 5.09 (d, J = 17.1 Hz, 1H), 4.39 (ddd, J = 21.1, 1Ό.2, 5.7 Hz, 2H), 4.23 (dd, J = 21.7, 5.8 Hz, 1H), 4.16 - 4.02 (m, 1H), 3.86 (ddd, J = 37.2,

12.9, 3.3 Hz, 1H), 2.82 - 2.47 (m, 1H), 2.39 (s, 3H), 2.16 - 1.91 (m, 1H); ¹⁹F NMR (282

MHz, DMSO-dô) δ -121.32, -121.66, -176.15, -176.34; MS (ES+): 463.0 (M+l), (ES-): 461.0 (M-l); Analysis calculated for C₂iH₂iClF₂N6O₂-HCl-2.25H₂O: C, 46.72; H, 4.95; Cl, 13.13; N, 15.57; Found: C, 46.35; H, 4.56; Cl, 13.52; N, 15.40.

Scheme 151

288

ο νη₂

Préparation of 4-amino-l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylic acid (151a)

Compound 151a was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylate (147a) (22 mg, 0.043 mmol) in THF (3 mL) and MeOH (3 mL) using a solution of lithium hydroxide (18.62 mg, 0.435 mmol) in water (3 mL) and stirring at RT for 19 h. Additional lithium hydroxide hydrate (25 mg, and 50 mg), 2N sodium hydroxide solution (0.4 mL and 0.6 mL) were added and stirred at RT overnight after each addition. This gave after workup and purification by reverse phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] 4-amino-l-(2-((2S,4R)-2-((3-chloro-2fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5carboxylic acid (151a) (2.2 mg, 10% yield) HCl sait as a white solid; ’H NMR (300 MHz,

DMSO-ifc) (a mixture of two rotamers) δ 8.64 (t, J = 5.7 Hz, 1H, D2O exchangeable), 8.53 & 8.48 (2s, 1H), 7.55 - 6.92 (m, 5H), 5.62 - 5.14 (m, 3H), 4.97 - 3.70 (m, 5H), 2.68 - 2.38 (m, 1H), 2.27 (s, 1H); ¹⁹F NMR (282 MHz, DMSO-d₆) (a mixture of two rotamers) δ -121.32 & 121.65 (1F), -175.81 & -176.07 (1F); MS (ES+): 492.00 & 494.00 (M+l); MS (ES-): 490.00 & 492.00 (M-l).

₂q Scheme 152

Préparation of (2S,4R)-l-(2-(6-amino-8-methyl-9H-purin-9-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fhioropyrrolidine-2-carboxamide (152d)

Step-1: Préparation of tert-butyl 2-(6-amino-8-methyl-9H-purin-9-yl)acetate (152b)

Compound 152b was prepared according to the procedure reported in step-1 of scheme-1, from 8-methyl-7H-purin-6-amine (152a) (500 mg, 3.35 mmol; CAS # 22387-37-7) in DMF (15 mL) using tert-butyl 2-bromoacetate and K2CO3 (556 mg, 4.02 mmol). This gave after workup tert-butyl 2-(6-amino-8-methyl-9H-purin-9-yl)acetate (152b) (390 mg, 44 % yield) as a yellow solid; 1H NMR (300 MHz, DMSO-J6) δ 8.07 (s, 1H), 7.11 (s, 2H), 4.90 (s, 2H),

2.42 (s, 3H), 1.42 (s, 9H); MS (ES+): 264.10 (M+l).

Step-2: Préparation of 2-(6-amino-8-methyl-9H-purin-9-yl)acetic acid (152c)

Compound 152c was prepared according to the procedure reported in step-2 of scheme-1, i from tert-butyl 2-(6-amino-8-methyl-9H-purin-9-yl)acetate (152b) (150 mg, 0.570 mmol) in

DCM (10 mL) using TFA and stirred at RT. This gave after workup 2-(6-amino-8-methyl75 9H-purin-9-yl)acetic acid (152c) which was used as such in the next step; MS (ES+): 208.10 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(6-amino-8-methyl-9H-purin-9-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (152d)

I I

Compound 152d was prepared according to the procedure reported in step-3 of scheme-1, 20 from TFA sait of 2-(6-amino-8-methyl-9H-purin-9-yl)acetic acid (152c) (118 mg, 0.57

I ! i i

290 mmol) in DMF (10 mL) using TFA sait of (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (7a) (188 mg, 0.684 mmol), HATU (433 mg, 1.140 mmol), DIPEA (0.496 mL, 2.85 mmol) and stirring at RT overnight. This gave after workup and purification by flash column chromatography [silica gel, eluting with DCM/methanol (1:0 to

19:1)] (2S,4R)-l-(2-(6-amino-8-methyl-9H-purin-9-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)4-fluoropyrrolidine-2-carboxamide (152d) (31 mg, 12% yield) as a white solid; 'H NMR (300 MHz, DMSO-de) (a mixture of two rotamers) δ 9.00 & 8.64 (2t, J - 6.2 Hz, 1H), 8.22 & 8.20 (2s, 1H), 7.54 - 7.39 (m, 1H), 7.31 - 7.17 (m, 1H), 7.11 - 7.00 (m, 1H), 5.62 - 5.36 (m, 1H), 5.31 (d, J = 17.5 Hz, 1H), 5.03 (d, J = 17.6 Hz, 1H), 4.61 - 3.74 (m, 5H), 2.60-1.90 (m,

2H), 2.37 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -121.28, -175.82; MS (ES+): 464.0 (M+l).

Scheme 153

Préparation of 4-amino-7-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-415 fluoropyrrolidin-l-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylic acid (153a)

Compound 153a was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-7-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-4fluoropyrrolidin-l-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylate (146a) (15 mg, 0.030 mmol) in THF (3 mL) and MeOH (3 mL) using a solution of lithium hydroxide (25.3 mg, 0.592 mmol) in water (3 mL) and stirring at RT overnight. This gave after workup and purification by reverse phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] 4-amino-7-(2-((2S,4R)-2-((3-chloro-2fluorobenzyl)carbamoyl)-4-fluoropyrrolidin-l-yl)-2-oxoethyl)-7H-pyrrolo[2,3-d]pyrimidine5-carboxylic acid (153a) (4.6 mg, 32%) HCl sait as a white solid; 'H NMR (300 MHz,

DMSO-de) (a mixture of two rotamers) δ 9.08 & 8.65 (2t, J = 5.9 Hz, 1H, D2O

291 exchangeable), 8.49 (s, 2H, D2O exchangeable), 8.38 & 8.36 (2s, 1H), 8.11 & 8.10 (2s, 1H), 7.52 - 7.01 (m, 3H), 5.66 - 5.04 (m, 3H), 4.89 - 3.70 (m, 5H), 2.75 - 2.30 (m, 1H), 2.23 1.89 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-îL) (a mixture of two rotamers) δ -121.34 & 121.63 (1F), -176.12 & -176.34 (1F); MS (ES+): 493.00 & 495.00 (M+l); MS (ES-): 491.00 & 493.00 (M-l).

Scheme 154

154a

154b

154c

154d

154e

Préparation of (lR,3S,5R)-2-(2-(4-amino-5-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (154e)

Step-1: Préparation of 5-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (154b)

Compound 154b was prepared according to the procedure reported in step-4 of scheme-130, from 4-chloro-5-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine (154a) (3.96 g, 19.83 mmol; CAS # 144927-56-0)) in isopropanol (20 mL) using ammonium hydroxide (11.26 g, 198 mmol; 30 wt.% in water) in a sealed tube and heating at 100 °C for 16 h. This gave after workup and 75 purification by flash column chromatography [S1O2 (24 g), eluting with MeOH in DCM from

0-3%] 5-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (154b) (1.94 g, 54.3 % yield) as a yellow solid, Ή NMR (300 MHz, DMSO-d₆) δ 11.83 (s, 1H), 8.06 (s, 1H), 7.31 (s, 1H), 6.75 (s, 2H), 2.32 (s, 3H); MS (ES+): 181 (M+l).

Step-2: Préparation of ieri-butyl 2-(4-amino-5-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-720 yl)acetate (154c)

Compound 154c was prepared according to the procedure reported in step-1 of scheme-1, from 5-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (154b) (1.94 g, 10.76 mmol) in

292

DMF (20 mL) using ZerZ-butyl 2-bromoacetate (2.100 g, 10.76 mmol), CS2CO3 (4.21 g, 12.92 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] ZerZ-butyl 2-(4amino-5-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (154c) (2.73 g, 86% yield) as 5 a pale orange solid; Ή NMR (300 MHz, DMSO-d₆) δ 8.09 (s, 1H), 7.37 (s, 1H), 6.83 (s, 2H), 4.87 (s, 2H), 2.34 (d, J = 1.0 Hz, 3H), 1.41 (s, 9H); MS (ES+) 295 (M+l).

Step-3: Préparation of 2-(4-amino-5-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (154d)

Compound 154d was prepared according to the procedure reported in step-2 of scheme-1, 10 from ZerZ-butyl 2-(4-amino-5-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (154c) (2.73 g, 9.27 mmol) in DCM (20 mL) using TFA (106 g, 185 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] 2-(4-amino-5-(methylthio)-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetic acid (154d) (4.50 g) TFA sait as a pale-orange solid. ^!H NMR (300 15 MHz, DMSO-dô) δ 8.37 (s, 1H), 7.64 (s, 1H), 5.00 (s, 2H), 2.40 (s, 3H); ¹⁹F NMR (282 MHz,

DMSO-d₆) δ -74.22; MS (ES+): 239 (M+l), (ES-): 237 (M-l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-5-(methylthio)-7H-pyrrolo[2,3d]pyrimidin-7-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (154e)

Compound 154e was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-5-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (154d) (80 mg, 0.227 mmol) in DMF (5 mL) using (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (72.3 mg, 0.227 mmol), HATU (104 mg, 0.273 mmol), DIPEA (147 mg, 1.135 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), using MeOH in DCM from 0 to 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-5-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (154e) (46 mg, 40% yield)

HCl sait as a white solid; ^XH NMR (300 MHz, DMSO-d₆) δ 10.80 (s, 1H, D₂O exchangeable), 8.41 (s and bs, 3H, 2H D2O exchangeable), 8.02 (d, J = 8.2 Hz, 1H), 7.72 (t, J = 8.0 Hz, 1H), 7.63 (s, 1H), 7.33 (d, J = 7.7 Hz, 1H), 5.50 (d, J = 17.0 Hz, 1H), 5.20 (d, J = 17.0 Hz, 1H), 4.43 (dd, J = 9.0, 5.5 Hz, 1H), 3.74 (ddd, J = 7.2, 5.3, 2.2 Hz, 1H), 2.39 (s, 3H), 2.30 (dd, J =

293 .

13.6, 9.2 Hz, 1H), 2.19 (dt, J = 12.9, 5.9 Hz, 1H), 1.95 - 1.81 (m, 1H), 1.06 - 0.95 (m, 1H), 0.73-0.62 (m, 1H); MS (ES+): 502.0 (M+l); (ES-): 500.0 (M-l); Analysis calculated for C2oH₂₀BrN₇0₂S.l.l.HC1.2.H₂0: C, 41.52; H, 4.37; Cl, 6.74; N, 16.95; Found: C, 41.36; H, 4.35; Cl, 6.99; N, 16.87.

Scheme 155

Préparation of 4-amino-l-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo [3.1,0]hexan-2-yl)-2-oxoethyl)-lH-pyrrolo [2,3-b]pyridine-5 -carboxylic acid (155f)

Step-1 : Préparation of methyl l-(2-(tert-butoxy)-2-oxoethyl)-4-chloro-lH-pyrrolo[2,310 b]pyridine-5-carboxylate (155b)

Compound 155b was prepared according to the procedure reported in step-1 of scheme-1, from methyl 4-chloro-lH-pyrrolo[2,3-b]pyridine-5-carboxylate (155a) (450 mg, 2.137 mmol; CAS # 951625-93-7) in DMF (10 mL) using tert-butyl 2-bromoacetate (0.379 mL, 2.56 mmol), Cs₂CO₃ (835 mg, 2.56 mmol) and stirring at RT for 20 h. This gave after workup and 75 purification by flash column chromatography [silica gel, eluting with EtOAc in Hexane from 0-14%] methyl l-(2-(Zert-butoxy)-2-oxoethyl)-4-chloro-lH-pyrrolo[2,3-b]pyridine-5carboxylate (155b) (605 mg, 87% yield) as a white solid; ³H NMR (300 MHz, DMSO-de) δ 8.73 (s, 1H), 7.77 (d, J = 3.7 Hz, 1H), 6.71 (d, J = 3.6 Hz, 1H), 5.09 (s, 2H), 3.89 (s, 3H), 1.40 (s, 9H); MS (ES+): 325.00 (M+l).

294

Step-2: Préparation of methyl l-(2-(teri-butoxy)-2-oxoethyl)-4-((tertebutoxycarbonyl)amino)lH-pyrrolo[2,3-b]pyridine-5-carboxylate (155c)

Compound 155c was prepared according to the procedure reported in step-3 of scheme-17, from methyl l-(2-(tert-butoxy)-2-oxoethyl)-4-chloro-lH-pyrrolo[2,3-b]pyridine-55 carboxylate (155b) (560 mg, 1.724 mmol) in toluene (16 mL) using XPhos (82 mg, 0.172 mmol), t-butyl carbamate (303 mg, 2.59 mmol), Pd₂(dba)a (79 mg, 0.086 mmol), césium carbonate (562 mg, 1.724 mmol) and heating at 90 °C for 16 h. This gave after work up and purification by flash column chromatography [silica gel, eluting with EtOAc in Hexane from 0-17%] methyl l-(2-(tert-butoxy)-2-oxoethyl)-4-((tert-butoxycarbonyl)amino)-lH10 pyrrolo[2,3-b]pyridine-5-carboxylate (155c) (628 mg, 90% yield); NMR (300 MHz, DMSO-î/ô) δ 10.19 (s, 1H), 8.56 (s, 1H), 7.48 (d, J = 3.7 Hz, 1H), 6.78 (d, J = 3.6 Hz, 1H), 5.01 (s, 2H), 3.81 (s, 3H), 1.49 (s, 9H), 1.40 (s, 9H); MS (ES+): 406.10 (M+l).

Step-3: Préparation of 2-(4-amino-5-(methoxycarbonyl)-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (155d)

Compound 155d was prepared according to the procedure reported in step-2 of scheme-1, from methyl l-(2-(tert-butoxy)-2-oxoethyl)-4-((tert-butoxycarbonyl)amino)-lH-pyrrolo[2,3b]pyridine-5-carboxylate (155c) (0.620 g, 1.529 mmol) using TFA. This gave after work up 2-(4-amino-5-(methoxycarbonyl)-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (155d) (840 mg) which was used as such for the next step; MS (ES+): 250.00 (M+l).

Step-4: Préparation of methyl 4-amino-l-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5carboxylate (155e)

Compound 155e was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-5-(methoxycarbonyl)-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (155d) (95 mg, 0.383 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (122 mg, 0.383 mmol), HATU (291 mg, 0.766 mmol), DIPEA (0.334 mL, 1.915 mmol) and stirring at RT for 20 h. This gave after workup and purification by flash column chromatography [silica gel, MeOH in DCM from 0 - 5%] methyl 4-amino-l-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-230 azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylate(155e) (115 mg, 59% yield) as a yellow solid; ^XH NMR (300 MHz, DMSO-î/ô) δ 10.76 (s, 1H), 8.49 (s, 1H), 8.03 (d, J = 8.2 Hz, 1H), 7.71 (t, J = 7.9 Hz, 1H), 7.59 (s, 2H), 7.32 (d, J = 7.7 Hz,

295

1H), 7.14 (d, J = 3.5 Hz, 1H), 6.75 (d, J = 3.6 Hz, 1H), 5.38 (d, J = 17.0 Hz, 1H), 5.15 (d, J = 17.0 Hz, 1H), 4.42 (dd, J = 9.0, 5.6 Hz, 1H), 3.80 (s, 3H), 3.77 - 3.66 (m, 1H), 2.36 - 2.07 (m, 2H), 1.94 - 1.68 (m, 1H), 1.08 - 0.91 (m, 1H), 0.76 - 0.50 (m, 1H); MS (ES+): 513.00 (M+l).

Step-5: Préparation of 4-amino-l-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylic acid (155f)

Compound 155f was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-l-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylate (155e) 10 (110 mg, 0.214 mmol) in THF (2 mL) and acetonitrile (2 mL) using a solution of lithium hydroxide hydrate (27.5 mg, 0.643 mmol) in water (2 mL) and stirring at RT for 20 h. This gave after workup and purification by reverse phase column chromatography [C18 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] 4-amino-l-(2((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-275 oxoethyl)-lH-pyrrolo[2,3-b]pyridine-5-carboxylic acid (155f) (10 mg, 9% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) (a mixture of two rotamers) δ 10.88 & 10.77 (2s, 1H, D₂O exchangeable), 8.60 & 8.60 (2s, 1H), 8.07 — 7.93 (m, 1H), 7.78 — 7.65 (m, 1H), 7.42 - 7.21 (m, 2H), 7.05 - 6.93 (m, 1H), 5.75-5.15 (m, 2H), 4.92 & 4.43 (2dd, J = 8.9, 5.7 Hz, 1H), 3.82-3.62 (m, 1H), 2.43 - 1.63 (m, 3H), 1.34 - 0.63 (m, 2H); MS (ES+): 498.90 & 20 500.90 (M+l); MS (ES-): 496.90 & 498.90 (M-l).

296

Préparation of (lR,3S,5R)-2-(2-(4-amino-5-formyl-lH-pyrrolo[2,3-b]pyridin-l-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (156e)

Step-1: Préparation of tert-butyl 2-(4-chloro-5-(hydroxymethyl)-lH-pyrrolo[2,3-b]pyridin-l. yl)acetate (156b)

Compound 156b was prepared according to the procedure reported in step-1 of scheme-1, from (4-chloro-lH-pyrrolo[2,3-b]pyridin-5-yl)methanol (156a) (115 mg, 0.630 mmol; CAS # 1015610-07-7) in DMF (6 mL) using teri-butyl 2-bromoacetate (0.112 mL, 0.756 mmol), Cs₂CO₃ (246 mg, 0.756 mmol) and stirring at RT for 13 h. This gave after workup and purification by flash column chromatography [silica gel, eluting with EtOAc in Hexane from 10 0-50%] tert-butyl 2-(4-chloro-5-(hydroxymethyl)-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (156b) (155 mg, 83% yield) as a white solid; ^XH NMR (300 MHz, DMSO-Tû δ 8.30 (s, 1H), 7.63 (d, J = 3.6 Hz, 1H), 6.53 (d, J = 3.5 Hz, 1H), 5.30 (t, J = 5.6 Hz, 1H), 5.03 (s, 2H), 4.69 (d, J = 5.5 Hz, 2H), 1.40 (s, 9H); MS (ES+): 297.00 (M+l).

Step-2: Préparation of Zeri-butyl 2-(4-((teri-butoxycarbonyl)amino)-5-formyl-lH-pyrrolo[2,315 b]pyridin-l-yl)acetate (156c)

Compound 156c was prepared according to the procedure reported in step-3 of scheme-17, from Zeri-butyl 2-(4-chloro-5-(hydroxymethyl)-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (156b) (132 mg, 0.445 mmol) in toluene (10 mL) using XPhos (21.21mg, 0.044 mmol), t-butyl carbamate (78 mg, 0.667 mmol), Pd₂(dba)₃ (20.37 mg, 0.022 mmol), césium carbonate (145 20 mg, 0.445 mmol) and heating at 90 °C for 12 h. This gave after work up and purification by flash column chromatography [silica gel, eluting with EtOAc in Hexane from 0-20%] tertbutyl 2-(4-((ieri-butoxycarbonyl)amino)-5-formyl-lH-pyrrolo[2,3-b]pyridin-l-yl)acetate (156c) (50 mg, 30% yield); Ή NMR (300 MHz, DMSO-d₆) δ 10.76 (s, 1H), 10.00 (s, 1H), 8.58 (s, 1H), 7.50 (d, J = 3.7 Hz, 1H), 6.94 (d, J = 3.7 Hz, 1H), 5.04 (s, 2H), 1.52 (s, 9H), 25 1.41 (s, 9H); MS (ES+): 376.10 (M+l).

Step-3: Préparation of 2-(4-amino-5-formyl-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (156d)

Compound 156d was prepared according to the procedure reported in step-2 of scheme-1, from teri-butyl 2-(4-((ieri-butoxycarbonyl)amino)-5-formyl-lH-pyrrolo[2,3-b]pyridin-lyl)acetate (156c) (45 mg, 0.12 mmol) in DCM (10 mL) using TFA. This gave after work up 30 2-(4-amino-5-formyl-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (156d), which was used as such for the next step; MS (ES+): 220.00 (M+l).

297

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-5-formyl-lH-pyrrolo[2,3-b]pyridin-lyl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (156e)

Compound 156e was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-5-formyl-lH-pyrrolo[2,3-b]pyridin-l-yl)acetic acid (156d) (0.12 mmol) in

DMF (6 mL) using TFA sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (0.144 mmol), HATU (91 mg, 0.240 mmol), DIPEA (0.105 mL, 0.600 mmol) and stirring at RT for 14 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0-5%] (lR,3S,5R)-2-(2-(4-amino-5-formyl-lH-pyrrolo[2,3-b]pyridin-l10 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (156e) (36 mg, 62% yield) as a light yellow solid; ^]H NMR (300 MHz, DMSO-dô) δ 10.76 (s, 1H, D2O exchangeable), 9.84 (s, 1H), 8.25 (s, 1H), 8.03 (d, J = 8.2 Hz, 1H), 7.72 (t, J = 7.9 Hz, 1H), 7.33 (d, J = 7.7 Hz, 1H), 7.16 (d, J = 3.6 Hz, 1H), 6.77 (d, J = 3.6 Hz, 1H), 5.45 - 5.12 (m, 2H), 4.43 (dd, J = 9.0, 5.5 Hz, 1H), 3.82 - 3.65 (m, 1H), 2.35 - 2.10 (m, 2H), 1.93 - 1.78 (m,

1H), 1.27 - 1.19 (m, 1H), 1.08 - 0.94 (m, 1H); MS (ES+): 482.900 & 485.00 (M+l); MS (ES-): 480.90 & 483.00 (M-l).

Scheme 157

Préparation of (lR,3S,5R)-2-(2-(4-amino-5-(hydroxymethyl)-lH-pyrrolo[2,3-b]pyridin-l20 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-earboxamide (157a) .

To a solution of (lR,3S,5R)-2-(2-(4-amino-5-formyl-lH-pyrrolo[2,3-b]pyridin-l-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (156e) (23 mg, 0.048 mmol) in THF (4 mL) and MeOH (2 mL) was added sodium borohydride (5.51 mg, 0.143 mmol) stirred at RT for 19 h and diluted with ethyi acetate (100 mL), sat. NH4CI (50 mL).

The organic phase was washed with brine (50 mL), dried, filtered and concentrated in vacuum. The residue obtained was purified by flash column chromatography [silica gel (4 g),

298 eluting with MeOH in DCM (0 - 5%)] to give (lR,3S,5R)-2-(2-(4-amino-5-(hydroxymethyl)lH-pyrrolo[2,3-b]pyridin-l-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (157a) (12 mg, 52% yield) as a white solid; ^XH NMR (300 MHz, DMSO-dô) δ 10.74 (s, 1H, D₂O exchangeable), 8.03 (d, J = 8.2 Hz, 1H), 7.78 - 7.61 (m, 2H), 7.32 (d, J = 5 7.7 Hz, 1H), 7.06 (d, J = 3.5 Hz, 1H), 6.55 (d, J = 3.6 Hz, 1H), 5.99 (s, 2H, D₂O exchangeable), 5.29 (d, J = 17.0 Hz, 1H), 5.10 (d, J = 16.9 Hz, 1H), 4.82 (t, J = 5.2 Hz, 1H), 4.48 (d, J = 5.0 Hz, 2H), 4.42 (dd, J = 9.0, 5.5 Hz, 1H), 3.76 - 3.68 (m, 1H), 2.36 - 2.08 (m, 2H), 1.92 -1.72 (m, 1H), 1.05 - 0.92 (m, 1H), 0.70 - 0.56 (m, 1H); MS (ES+): 484.95 & 487.00 (M+l); MS (ES-): 482.95 & 484.95 (M-l).

Scheme 158

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (158e)

Step-1: Préparation of ethyl 2-(4-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate 15 (158b)

Compound 158b was prepared according to the procedure reported in step-1 of scheme-1, from 4-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidine (158a) (1 g, 5.97 mmol; CAS # 3580868-5) in DMF (25 mL) using ethyl 2-bromoacetate (0.794 mL, 7.16 mmol), Cs₂CO3 (2.33 g, 7.16 mmol) and stirring at 0 °C for 20 min. This gave after workup and purification by flash 20 column chromatography [silica gel, eluting with EtOAc in Hexane from 0-25%] ethyl 2-(4chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (158b) (1.32 g, 87% yield) as a light yellow solid; Ή NMR (300 MHz, DMSO-d₆) δ 8.56 (s, 1H), 6.51 - 6.46 (m, 1H), 5.17

299 (s, 2H), 4.16 (q, J = 7.1 Hz, 2H), 2.43 (d, J = 1.1 Hz, 3H), 1.21 (t, J = 7.1 Hz, 3H); MS (ES+): 254.00 (M+l).

Step-2: Préparation of ethyl 2-(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (158c)

Compound 158c was prepared according to the procedure reported in step-3 of scheme-17, from ethyl 2-(4-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (158b) (500 mg, 1.971 mmol) in toluene (15 mL) using XPhos (94 mg, 0.197 mmol), ί-butyl carbamate (346 mg, 2.96 mmol), Pd₂(dba)₃ (90 mg, 0.099 mmol), césium carbonate (642 mg, 1.971 mmol) and heating at 90 °C for 13 h under nitrogen. This gave after work up and purification by flash column chromatography [silica gel, eluting with EtOAc in Hexane/10% MeOH from 050%] ethyl 2-(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (158c) (110 mg, 24% yield); MS (ES+): 235.10 (M+l).

Step-3: Préparation of 2-(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (158d)

Compound 158d was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (158c) (100 mg, 0.427 mmol) in THF (3 mL) and MeOH (3 mL) using a solution of lithium hydroxide hydrate (110 mg, 2.56 mmol) in water (3 mL) and stirring at RT for 19 h. This gave after work up 2(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (158d) (88 mg,100% yield) which was used as such for the next step; MS (ES+): 207.10 (M+l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (158e)

Compound 158e was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid (158d) (0.427 mmol) in DMF (15 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (163 mg, 0.512 mmol), HATU (325 mg, 0.854 mmol), DIPEA (0.372 mL, 2.135 mmol) and stirring at RT for 23 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0-5 %] followed by conversion of product obtained to HCl sait by dissolving in acetonitrile (2 mL) and 50 mM aq. HCl (8 mL) and lyophilization (lR,3S,5R)-2-(2-(4-amino6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetyl)-N-(6-bromopyridin-2-yl)-221436

300 azabicyclo[3.1.0]hexane-3-carboxamide (158e) (50 mg, 25% yield) as an off white solid; ¹H NMR (300 MHz, DMSO-t/ô) δ 10.80 (s, 1H, D2O exchangeable), 8.33 (s, 1H), 8.02 (d, J = 8.1 Hz, 1H), 7.72 (t, J = 8.0 Hz, 1H), 7.33 (d, J = 7.7 Hz, 1H), 6.68 (d, J = 1.3 Hz, 1H), 5.47 (d, J = 17.4 Hz, 1H), 5.20 (d, J = 17.3 Hz, 1H), 4.43 (dd, J = 9.1, 5.5 Hz, 1H), 3.83 (td, J =

6.7, 6.2, 2.3 Hz, 1H), 2.42 - 2.12 (m, 5H), 1.95 - 1.77 (m, 1H), 1.12 - 0.93 (m, 1H), 0.78 0.59 (m, 1H); MS (ES+): 470.10 & 472.10 (M+l); MS (ES-): 468.10 & 470.10 (M-l); Analysis calculated for C2oH₂oBrN70₂.1.2HC1.2.5H20: C, 42.96; H, 4.72; Cl, 7.61; N, 17.54; Found: C, 42.96; H, 4.48; Cl, 7.38; N, 17.36.

Scheme 159

OMe

38e

159a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (159a)

Compound 159a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (38e) (100 mg, 0.259 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (95 mg, 0.285 mmol), HATU (148 mg, 0.388 mmol), DIPEA (167 mg, 1.294 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0-3 %] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (159a) (117 mg, 82% yield) HCl sait as a white solid; ¹H NMR (300 MHz, DMSO-dâ) δ 10.77 (s, 1H, D2O exchangeable), 8.75 (s, 2H, D2O exchangeable), 8.60 (s, 1H), 8.08 (d, J = 2.3 Hz, 1H),

8.01 (d, J = 8.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.62 (d, J = 9.0 Hz, 1H), 7.32 (d, J = 7.7 Hz,

301

1H), 7.19 (dd, J = 9.1, 2.3 Hz, 1H), 5.69 (d, J = 17.3 Hz, 1H), 5.35 (d, J = 17.3 Hz, 1H), 4.37 (dd, J = 9.1, 5.9 Hz, 1H), 3.88 (s, 3H), 3.71 - 3.66 (m, 1H), 2.44 (s, 1H), 1.98 (dd, J = 13.3, 5.8 Hz, 1H), 1.31 (s, 3H), 1.07 - 0.98 (m, 1H), 0.98 - 0.83 (m, 1H); MS (ES+): 550 (M+l), (ES-): 548 (M-l); Analysis calculated for C2sH24BrN7O3.HC1.2.25H2O: C, 47.86; H, 4.74; Cl, 5 5.65; N, 15.63; Found: C, 47.64; H, 4.71; Cl, 5.77; N, 15.34.

Scheme 160

Br

34d

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (160a)

Compound 160a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (34d) (50 mg, 0.115 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (42 mg, 0.126 mmol), HATU (65.5 mg, 0.172 mmol), DIPEA (74.3 mg, 0.575 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), MeOH in DCM from 0-3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] to afford (lR,3S,5R)-2-(2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (160a) (42 mg, 61% yield) HCl sait as a white solid; ³H NMR (300 MHz, DMSO-rfâ) δ 10.77 (s, 1H, D2O exchangeable), 8.96 - 8.69 (m, 3H, 2H D2O exchangeable), 8.65 (s, 1H), 8.01 (d, J = 8.1 Hz, 1H), 7.77 - 7.64 (m, 3H), 7.32 (d, J = 7.7 Hz, 1H), 5.73 (d, J = 17.4 Hz, 1H), 5.37 (d, J = 17.3 Hz, 1H), 4.37 (dd, J = 9.1, 5.9 Hz, 1H), 3.69 - 3.65 (m, 1H), 2.45 (d, J = 8.9 Hz, 1H), 1.98 (dd, J = 13.3, 5.9 Hz, 1H), 1.31 (s, 3H), 1.01 (m, 1H), 0.93 (m, 1H); MS (ES+): 598 (M+l), (ES-): 596 (M-l); Analysis calculated for C24H21Br2N7O2.HCl.L5.H2O: C, 43.49; H, 3.80; Cl,

5.35; N, 14.79; Found: C, 43.36; H, 3.76; Cl, 5.49; N, 14.69.

302

Scheme 161

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxylic acid (161a)

Compound 161a was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxylate (40a) (107 mg, 0.185 mmol) in THF (3 mL) and using 2M aqueous lithium hydroxide hydrate (0.111 mL, 0.222 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5b]indole-6-carboxylic acid (161a) (33 mg, 32% yield) HCl sait as a white solid; ³H NMR (300 MHz, DMSO-î/ô) δ 10.77 (s, 1H, D2O exchangeable), 9.10 (s, 1H), 8.81 - 8.40 (m, 3H,

2H D2O exchangeable), 8.11 (dt, J = 8.8, 2.0 Hz, 1H), 7.97 (dd, J = 18.4, 8.2 Hz, 1H), 7.70 (ddd, J = 16.5, 8.4, 3.9 Hz, 2H), 7.30 (dd, J = 7.8, 5.2 Hz, 1H), 5.74 (d, J = 17.8 Hz, 1H), 5.39 (dd, J = 17.2, 3.4 Hz, 1H), 4.37 (dd, J = 9.0, 5.9 Hz, 1H), 3.70 - 3.68 (m, 1H), 2.45 2.33 (m, 1H), 2.12 - 1.90 (m, 1H), 1.28 (s, 3H), 1.07 - 0.80 (m, 2H); MS (ES+) 564.0 (M+l);

(ES-): 562.0 (M-l).

Scheme 162

303

O₂N

NC^CN

162a

NaH

162b

Zn

AcOH

H NH₂ · AcOH

NH

NH₄OAc, AcOH HC(OMe)₃

H N

NH₂

162d

162c ⁱBuO₂C Br

TFA

N

Cs₂CO₃

162e

Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[3',2^,:4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (162g)

Step-1: Préparation of 2-(2-nitropyridin-3-yl)malononitrile (162b)

NaH (1.958 g, 49.0 mmol) was added portion-wise to a cold solution of malononitrile (3.23 g, 49.0 mmol) in THF (50 mL) at 0 °C, during which hydrogen gas liberated and a white precipitate formed. The resulting cloudy white mixture was stirred at 0 °C for 1 h, followed by addition of 3-bromo-2-nitropyridine (162a) (4.92 g, 24.24 mmol). The resulting deep red/black suspension was then heated at 60 °C under argon for 3 h. The cooled reaction was quenched with saturated NH4CI (50 mL) and diluted with EtOAc (50 mL). After 30-min stirring at rt, the two layers were separated. The aqueous layer was extracted with EtOAc (30 mL x 3). The combined extract was concentrated to provide the crude product 2-(2nitropyridin-3-yl)malononitrile (162b) (5.75 g,) as a brown/black solid, which was used as such in the next reaction.; MS (ES+): 159 (M+l), (ES-): 157 (M-l).

Step-2: Préparation of 2-amino-lH-pyrrolo[2,3-b]pyridine-3-carbonitrile (162c)

Crude 2-(2-nitropyridin-3-yl)malononitrile (162b) (5.75 g) from step-1 above was diluted in acetic acid (36.7 g, 611 mmol), followed by slow addition of zinc (10.00 g, 153 mmol) at rt. The mixture was stirred at 60 °C for 2 h and then filtered hot. The filtered cake was washed

304 thoroughly with boiling EtOH. The filtrate was concentrated to dryness. The residue was suspended in H₂O (50 mL), neutralized with 3 M aqueous NaOH until pH 7, and then extracted with EtOAc (50 mL x 3). The combined extract was washed with H₂O (30 mL x 2), brine (30 mL), dried, filtered and concentrated in vacuum. The residue obtained was purified by flash column chromatography (SiO₂, 40 g, eluting with 0-20% DMA80 in DCM) to provide 2-amino-lH-pyrrolo[2,3-b]pyridine-3-carbonitrile (162c) (1.00 g, 21 % yield) as a beige solid; 'HNMR (300 MHz, DMSO-î/₆) δ 11.43 (s, 1H), 7.89 (dd, J = 5.0, 1.5 Hz, 1H), 7.47 (dd, J = 7.7,1.5 Hz, 1H), 7.11 - 6.84 (m, 3H); MS (ES+): 159 (M+l), (ES-): 157 (M-l).

Step-3: Préparation of gH-pyridop^'^^lpyrrolop^-dJpyrimidin^-amine (162d)

Compound 162d was prepared according to the procedure reported m step-2 of scheme-29, from 2-amino-lH-pyrrolo[2,3-b]pyridine-3-carbonitrile (162c) (1.00 g, 6.32 mmol) m éthanol (30 mL) using formamidine acetate (5.27 g, 50.6 mmol), NH₄OAc (1.462 g, 18.97 mmol), glacial AcOH (1.139 g, 18.97 mmol), HC(OMe)₃ (6.71 g, 63.2 mmol) and heating at 80 °C for 5 days. The solid obtained was collected by filtration washed with boiling EtOH and air15 dried to provide 9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (162d) (0.27 g, 23% yield) as a brown solid; Ή NMR (300 MHz, DMSO-ifc) δ 12.36 (s, 1H), 8.68 (dd, J = 7.8, 1.6 Hz, 1H), 8.36 (dd, J = 4.9,1.5 Hz, 1H), 8.31 (s, 1H), 7.34 (s, 2H), 7.25 (dd, J = 7.8, 4.9 Hz, 1H); MS (ES+): 186 (M+l), (ES-): 184 (M-l).

Step-4: Préparation of teri-butyl 2-(4-amino-9H-pyrido[3^,,2':4,5]pyrrolo[2,3-d]pynmidin-920 yl)acetate (162e)

Compound 162e was prepared according to the procedure reported in step-1 of scheme-1, from 9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (162d) (0.27 g, 1.458 mmol) in DMF (10 mL) using teri-butyl 2-bromoacetate (0.341 g, 1.750 mmol) and Cs₂CO₃ (0.950 g, 2.92 mmol). This gave after workup and purification using flash column chromatography

[SiO₂ gel (24 g), eluting with MeOH in DCM from 0-4%] teri-butyl 2-(4-amino-9Hpyrido[3’,2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (162e) (0.27 g, 0.902 mmol, 62% yield) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-d₆) δ 8.75 (dd, J = 7.8, 1.5 Hz, 1H), 8.40 (dd, J = 4.9,1.5 Hz, 1H), 8.36 (s, 1H), 7.50 (s, 2H), 7.34 (dd, J = 7.8, 4.9 Hz, 1H), 5.06 (s, 2H), 1.40 (s, 9H); MS (ES+): 300 (M+l), (ES-): 298 (M-l).

Step-5: Préparation of 2-(4-amino-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (162f)

305

Compound 162f was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-9H-pyrido[3',2^,:4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (162e) (0.27 g, 0.902 mmol) in DCM (10 mL) using TFA (1.029 g, 9.02 mmol) and stirring at RT for 16 h. This gave after workup TFA sait of 2-(4-amino-9H-pyrido[3',2':4,5]pyrrolo[2,35 d]pyrimidin-9-yl)acetic acid (162f) (311 mg, 97 % yield) as a pale-yellow solid; ^]H NMR (300 MHz, DMSO-dô) δ 8.84 (dd, J = 7.9, 1.5 Hz, 1H), 8.59 - 8.44 (m, 2H), 8.04 (s, 2H), 7.43 (dd, J = 7.8, 4.9 Hz, 1H), 5.14 (s, 2H). ¹⁹F NMR (282 MHz, DMSO) δ -74.47; MS (ES+): 244 (M+l), (ES-): 242 (M-l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[3',2^,:4,5]pyrrolo[2,310 d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (162g)

Compound 162g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (162f) (75 mg, 0.210 mmol) in DMF (5 mL) using (lR,3S,5R)-N-(6-bromopyndm-2-yl)-275 azabicyclo[3.1.0]hexane-3-carboxamide (4a) (80 mg, 0.252 mmol), HATU (120 mg, 0.315 mmol), DIPEA (136 mg, 1.050 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH m DCM from 0-6%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[3',2';4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (162g) (75 mg, 70% yield) HCl sait as a white solid; ^XH NMR (300 MHz, DMSO-^) δ 10.78 (s, 1H, D₂O exchangeable), 9.29 - 8.91 (m, 3H, 2H D₂O exchangeable), 8.74 (s, 1H), 8.59 (dd, J = 4.8,1.4 Hz, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.53 (dd, J = 7.9, 4.8 Hz, 1H), 7.32 (d, J = 7.7

Hz, 1H), 5.78 (d, J = 17.0 Hz, 1H), 5.41 (d, J = 16.9 Hz, 1H), 4.42 (m, 1H), 4.03 - 3.84 (m, 1H), 2.39 - 2.27 (m, 1H), 2.27 - 2.18 (m, 1H), 1.99 -1.86 (m, 1H), 1.11 (m, 1H), 0.70 (m, 1H); MS (ES+): 507/509 (M+l), 505/507 (M-l).

Scheme 163

306

163e

NH₄OAc, AcOH HC(OMe)₃

Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[3^,,4':4,5]pyrrolo[2,3-d]pyrimidm-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (163g)

Step-1: Préparation of 2-(4-nitropyridin-3-yl)malononitrile (163b)

Compound 163b was prepared according to the procedure reported in step-1 of scheme-162 from 3-bromo-4-nitropyridine (163a) (5.00 g, 24.63 mmol) using NaH (1.990 g, 49.8 mmol) malononitrile (3.29 g, 49.8 mmol) in THF (100 mL). This gave after workup 2-(4nitropyridin-3-yl)malononitrile (163b) (5.27 g) as a brown/black solid, which was used as such in the next reaction; MS (ES+): 159 (M+l), (ES-): 157 (M-l).

Step-2: Préparation of 2-amino-lH-pyrrolo[3,2-c]pyridine-3-carbonitrile (163c)

Compound 163c was prepared according to the procedure reported in step-2 of scheme-162 from 2-(4-nitropyridin-3-yl)malononitrile (163b) (5.27 g, 28.0 mmol) in acetic acid (16.82 g, 280 mmol), using zinc (9.16 g, 140 mmol). This gave after workup and purification by flash column chromatography (SiO₂, 40 g, eluting with 0-20% DMA-80 in DCM) 2-ammo-lH75 pyrrolo[3,2-c]pyridine-3-carbonitrile (163c) (0.23 g, 1.454 mmol, 5.19 % yield) as a beige solid; Ή NMR (300 MHz, DMSO-ifc) δ 11.30 (s, 1H), 8.34 (d, J = 0.9 Hz, 1H), 8.04 (d, J =

307

5.4 Hz, 1H), 7.33 - 7.01 (m, 3H). LC-MS: t = 0.54 min; MS (ES+): 159 (M+l), (ES-): 157 (M-l).

Step-3: Préparation of 9H-pyrido[3',4^,:4,5]pyrrolo[2,3-d]pyrimidin-4-amine (163d)

Compound 163d was prepared according to the procedure reported in step-2 of scheme-29, 5 from 2-amino-lH-pyrrolo[3,2-c]pyridine-3-carbonitrile (163c) (0.23 g, 1.454 mmol) in éthanol (20 mL) using formamidine acetate (1.211 g, 11.63 mmol), NH4OAC (0.336 g, 4.36 mmol), glacial AcOH (0.262 g, 4.36 mmol), HC(OMe)3(0.772 g, 7.27 mmol) and heating at 80 °C for 5 days. This gave after work up 9H-pyrido[3',4':4,5]pyrrolo[2,3-d]pyrimidin-4amine (163d) (0.14 g, 52% yield) as a brown solid; ³H NMR (300 MHz, DMSO-î/ô) δ 12.24 10 (s, 1H), 9.47 (d, J = 1.0 Hz, 1H), 8.42 (d, J = 5.6 Hz, 1H), 8.32 (s, 1H), 7.42 (dd, J = 5.6,1.0

Hz, 1H), 7.37 (s, 2H). MS (ES+): 186 (M+l); (ES-): 184 (M-l).

Step-4: Préparation of ZerZ-butyl 2-(4-amino-9H-pyrido[3^,,4^,:4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetate (163e)

Compound 163e was prepared according to the procedure reported in step-1 of scheme-1, 15 from 9H-pyrido[3',4^,:4,5]pyrrolo[2,3-d]pyrimidin-4-amine (163d) (0.14 g, 0.756 mmol) in

DMF (10 mL) using ZerZ-butyl 2-bromoacetate (0.177 g, 0.907 mmol) and CS2CO3 (0.493 g, 1.512 mmol). This gave after workup and purification using flash column chromatography [S1O2 gel (24 g), eluting with MeOH in DCM from 0-6%] ZerZ-butyl 2-(4-amino-9Hpyridop'^'^^JpyrroloPjS-dJpyrimidin^-yljacetate (163e) (0.11 g, 49 % yield) as a pale20 yellow solid; Ή NMR (300 MHz, DMSO-d₆) δ 9.51 (s, 1H), 8.49 (d, J = 5.6 Hz, 1H), 8.37 (s, 1H), 7.64 (dd, J = 5.7, 0.9 Hz, 1H), 7.52 (s, 2H), 5.16 (s, 2H), 1.40 (s, 9H).

Step-5: Préparation of 2-(4-amino-9H-pyrido[3',4':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (1631)

Compound 163f was prepared according to the procedure reported in step-2 of scheme-1, 25 from ZerZ-butyl 2-(4-amino-9H-pyrido[3^,,4':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (163e) (0.108 g, 0.361 mmol) in DCM (10 mL) using TFA (411 mg, 3.61 mmol) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-9H-pyrido[3',4':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetic acid (163f) (202 mg) as a pale-yellow solid; ’H NMR (300 MHz, DMSO-dô) δ 9.87 (d, J = 2.1 Hz, 1H), 8.85 (dd, J = 6.9, 2.1 Hz, 1H), 8.55 (d, J = 2.1 Hz, 1H), 30 8.37 (dd, J = 6.7, 2.1 Hz, 1H), 8.10 (s, 2H), 5.37 (d, J = 2.0 Hz, 2H). ¹⁹F NMR (282 MHz,

DMSO-ife) δ -74.58; MS (ES+): 244 (M+l), (ES-): 242 (M-l).

308

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[3',4':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (163g)

Compound 163g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait 2-(4-amino-9H-pyrido[3',4':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (163f) (75 mg, 0.210 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (80 mg, 0.252 mmol), HATU (120 mg, 0.315 mmol), DIPEA (136 mg, 1.050 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0-7%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[3',4':4,5]pyrrolo[2,3-d]pyrimidin9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicycio [3.1,0]hexane-3-carboxamide (163g) (43 mg, 40% yield) HCl sait as a pale yellow solid; ³H NMR (300 MHz, DMSO-ifc) δ 10.78 (s,

1H, D2O exchangeable), 10.08 (s, 1H), 8.83 (d, J = 6.6 Hz, 1H), 8.62 (s, 1H), 8.44 (s, 2H,

D2O exchangeable),), 8.25 (d, J = 6.7 Hz, 1H), 8.00 (d, J = 8.1 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.92 (d, J = 17.4 Hz, 1H), 5.53 (d, J = 17.4 Hz, 1H), 4.48 - 4.37 (m, 1H), 3.97 - 3.83 (m, 1H), 2.37 - 2.29 (m, 1H), 2.28 - 2.17 (m, 1H), 1.99 - 1.85 (m, 1H), 1.15 - 1.02 (m, 1H), 0.91 - 0.78 (m, 1H); MS (ES+): 507/509 (M+l); (ES-): 505/507 (M-l).

Scheme 164

159a

164a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-hydroxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (164a)

To a solution of (lR,3S,5R)-2-(2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)25 N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (159a) (70 mg,

309

0.127 mmol) in DCM (5 mL) was added 1.0 M BBr₃ in DCM (0.636 mL, 0.636 mmol) drop wise at 0 °C under argon and stirred at RT for 16 h. The reaction mixture was quenched with Η₂Ο (2 mL) and concentrated in vacuum. The residue was purified by reverse-phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] to provide (lR,3S,5R)-2-(2-(4-amino-6-hydroxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (164a) (41 mg, 60% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-ifc) δ 10.77 (s, 1H, D₂O exchangeable), 9.46 (s, 1H, D₂O exchangeable), 8.54 (s, 1H), 8.46 (s, 2H, D2O exchangeable), 8.01 (d, J = 8.2 Hz, 1H), 7.84 (d, J = 2.2 Hz, 1H), 7.70 (t, J = 7.9 Hz, 1H),

ΊΑΊ (d, J = 8.8 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 7.06 (dd, J = 8.8, 2.3 Hz, 1H), 5.63 (d, J = 17.3 Hz, 1H), 5.29 (d, J = 17.3 Hz, 1H), 4.36 (m, 1H), 3.66 (dd, J = 5.4, 2.3 Hz, 1H), 2.52 2.39 (m, 1H), 1.97 (dd, J = 13.2, 5.9 Hz, 1H), 1.29 (s, 3H), 1.06 - 0.94 (m, 1H), 0.94 - 0.84 (m, 1H); MS (ES+): 536/538 (M+l), (ES-): 534/536 (M-l).

Scheme 165

165a

Préparation of 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6-yl)propanoic acid (165a)

Compounds 165a was prepared according to the procedure reported in step-4 of scheme-17, 20 from ethyl 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6-yl)propanoate(45d) (78 mg, 0.129 mmol) in THF (3 mL) using 2M aqueous lithium hydroxide (0.077 mL, 0.154 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1%

HCl) from 0-100%] 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6-yl)propanoicacid

310 (165a) (55 mg, 74 % yield) HCl sait as a white solid; ^JH NMR (300 MHz, DMSO-de) δ 10.77 (s, 1H, D₂O exchangeable), 8.59 (d, J = 4.5 Hz, 1H), 8.51 (d, J = 9.2 Hz, 2H, D₂O exchangeable), 8.43 — 8.34 (m, 1H), 7.98 (dd, J = 16.3, 8.2 Hz, 1H), 7.70 (td, J — 8.1, 6.0 Hz, 1H), 7.59 (dd, J = 8.4, 2.2 Hz, 1H), 7.44 (d, J = 8.7 Hz, 1H), 7.31 (dd, J = 7.8, 5.4 Hz, 1H), 5 5.74 (d, J = 17.3 Hz, 1H), 5.39 (dd, J = 17.3, 3.0 Hz, 1H), 4.48 - 4.35 (m, 1H), 4.05 - 3.85 (m, 1H), 3.01 (t, J = 7.8 Hz, 2H), 2.67 (dd, J = 8.6, 6.9 Hz, 2H), 2.40 - 2.27 (m, 1H), 2.27 2.14 (m, 1H), 1.99 - 1.87 (m, 1H), 1.28 -1.02 (m, 1H), 0.96 - 0.74 (m, 1H); MS (ES+): 578/580 (M+l), (ES-): 576/578 (M-l); Analysis calculated for C26H24BrN7O4.HCl.2H2O: C, 47.98; H, 4.49; Cl, 5.45; N, 15.06; Found: C, 47.70; H, 4.29; Cl, 5.69; N, 14.77.

m Scheme 166

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-hydroxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (166a)

Compound 166a was prepared according to the procedure reported in scheme-164, from (lR,3S,5R)-2-(2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (38e) (85 mg, 0.158 mmol) m DCM (5 mL) using a solution of 1.0 M BBr₃ in DCM (0.792 mL, 0.792 mmol) and stirring at RT for 30 min. This gave after work up and purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-hydroxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (166a) (30 mg, 36% yield) HCl sait as a white solid; ^XH NMR (300 MHz, DMSO-dô) δ 10.77 (s, 1H, D2O exchangeable), 9.91 - 9.13 (m, 1H, D₂O exchangeable), 8.68 (s, 2H, D₂O exchangeable), 8.60 (s, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.87 (d, J = 2.2 Hz, 1H), 7.71 (t, J = 7.9 Hz, 1H), 7.52 (d, J = 8.8 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 7.11 (dd, J = 8.8, 2.3 Hz, 1H), 5.70 (d, J = 17.3 Hz,

1H), 5.36 (d, J = 17.2 Hz, 1H), 4.42 (dd, J = 9.1, 5.5 Hz, 1H), 3.89 (td, J = 6.2, 5.4, 2.2 Hz,

311

1H), 2.39 - 2.26 (m, 1H), 2.26 - 2.14 (m, 1H), 1.98 - 1.84 (m, 1H), 1.12 - 0.98 (m, 1H), 0.83 - 0.69 (m, 1H); MS (ES+): 522/524 (M+l), 520/522 (M-l).

Scheme 167

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-hydroxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (167a)

Compound 167a was prepared according to the procedure reported in scheme-164, from (lR,3S,5R)-2-(2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (36e) (82 mg, 0.153 mmol) in

DCM (5 ml) using a solution of 1.0 M BBrs in DCM (0.764 mL, 0.764 mmol) and stirring at RT for 16 h. This gave after work up and purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-hydroxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (167a) (30 mg, 38% yield) HCl sait as a white solid; ¹H NMR (300 MHz, DMSO-ôfc) δ 10.79 (s, 1H, D₂O exchangeable), 9.97 (s, 1H, D₂O exchangeable), 8.53 (d, J = 3.1 Hz, 1H), 8.37 (s, 2H, D₂O exchangeable), 8.28 (d, J = 8.5 Hz, 1H), 8.01 (d, J = 8.1 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 6.97 (d, J = 2.0 Hz, 1H), 6.89 (dd, J = 8.6, 2.1 Hz, 1H), 5.66 (d, J = 17.3 Hz, 1H), 5.31 (d, J = 17.4 Hz, 1H), 4.42 (dd, J = 9.0, 5.5 Hz, 1H), 3.90 (td, J = 6.9, 6.3,

2.5 Hz, 1H), 2.38 - 2.29 (m, 1H), 2.26 - 2.15 (m, 1H), 1.98 - 1.87 (m, 1H), 1.12 -1.02 (m,

1H), 0.77-0.70 (m, 1H); MS (ES+): 522/524 (M+l), (ES-): 520/522 (M-l).

Scheme 168

Préparation of (lR,3S,5R)-2-(2-(6-acetamido-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (168g)

Step-1: Préparation of N-(4-acetamido-2-bromophenyl)-2,2,2-trifluoroacetamide (168b)

Compound 168b was prepared according to the procedure reported in step-1 of scheme-46, from N-(4-amino-3-bromophenyl)acetamide (168a) (0.88 g, 3.84 mmol; CAS # 860745-87-5) in DCM (30 mL) using triethylamine (0.661 g, 6.53 mmol), trifluoroacetic acid anhydride (1.210 g, 5.76 mmol) and stirring at RT for 3 h. This gave after workup N-(4-acetamido-2bromophenyl)-2,2,2-trifluoroacetamide (168b) (400 mg, 32% yield) as a yellow solid which was used as such for the next step; *H NMR (300 MHz, DMSO-î/ô) δ 11.18 (s, 1H), 10.25 (s, 1H), 8.09 (d, J = 2.2 Hz, 1H), 7.55 (dd, J = 8.6, 2.3 Hz, 1H), 7.37 (d, J = 8.6 Hz, 1H), 2.07 (s, 3H). ¹⁹F NMR (282 MHz, DMSO) δ -74.07; MS (ES+): 325/327 (M+l); (ES-): 323/325 (ΜΙ)·

Step-2: Préparation of N-(2-amino-3-cyano-lH-indol-5-yl)acetamide (168c)

Compound 168c was prepared according to the procedure reported in step-1 of scheme-11, from N-(4-acetamido-2-bromophenyl)-2,2,2-trifluoroacetamide (168b) (400 mg, 1.230 mmol) in DMSO (10 mL) using malononitrile (203 mg, 3.08 mmol), L-proline (56.7 mg,

313

0.492 mmol), Cul (46.9 mg, 0.246 mmol), a solution of K2CO3 (340 mg, 2.461 mmol) in water (10 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [SiOzgel (24 g), eluting with MeOH in DCM from 0-5 %] N-(2amino-3-cyano-lH-indol-5-yl)acetamide (168c) (120 mg, 46% yield) as a yellow solid; ’H

NMR (300 MHz, DMSO-d₆) δ 10.58 (s, 1H), 9.72 (s, 1H), 7.52 (d, J = 1.5 Hz, 1H), 7.01 (d, J = 1.3 Hz, 2H), 6.72 (s, 2H), 2.01 (s, 3H). LC-MS: t = 1.47 min; MS (ES+): 215 (M+l); (ES-): 213 (M-l).

Step-3: Préparation of N-(4-amino-9H-pyrimido[4,5-b]indol-6-yl)acetamide (168d)

Compound 168d was prepared according to the procedure reported in step-2 of scheme-29, from N-(2-amino-3-cyano-lH-indol-5-yl)acetamide (168c) (1.77 g, 8.26 mmol) in éthanol (30 mL) using formamidine acetate (6.88 g, 66.1 mmol), NH4OAC (1.911 g, 24.79 mmol), ' HC(OMe)₃(8.77 g, 83 mmol) and heating at 80 °C for 16 h. This gave after work up N-(4amino-9H-pyrimido[4,5-b]indol-6-yl)acetamide (168d) (1.48 g, 74% yield) as a tan solid; ’H

NMR (300 MHz, DMSO-d₆) δ 11.76 (s, 1H), 9.81 (s, 1H), 8.22 (d, J = 2.2 Hz, 2H), 7.46 15 7.31 (m, 2H), 7.09 - 6.90 (m, 2H), 2.06 (s, 3H); MS (ES+): 242 (M+l), (ES-): 240 (M-l).

Step-4: Préparation of tert-butyl 2-(6-acetamido-4-amino-9H-pyrimido[4,5-b]indol-9yl)acetate (168e)

Compound 168e was prepared according to the procedure reported in step-1 of scheme-1, from N-(4-amino-9H-pyrimido[4,5-b]indol-6-yl)acetamide (168d) (0.74 g, 3.07 mmol) in DMF (10 mL) using tert-butyl 2-bromoacetate (0.718 g, 3.68 mmol) and CS2CO3 (1.999 g, 6.13 mmol). This gave after workup and purification using flash column chromatography [SiO2 gel (40 g), eluting with MeOH in DCM from 0-6%] tert-butyl 2-(6-acetamido-4amino-9H-pyrimido[4,5-b]indol-9-yl)acetate (168e) (0.77 g, 71% yield) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-d₆) δ 9.86 (s, 1H), 8.28 (s, 2H), 7.56 - 7.40 (m, 2H), 7.14 (s, 2H), 5.09 (s, 2H), 2.07 (s, 3H), 1.41 (s, 9H); MS (ES+): 356 (M+l), (ES-): 354 (M-l).

Step-5: Préparation of 2-(6-açetamido-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (168f)

Compound 168f was prepared according to the procedure reported in step-2 of scheme-1, from ieri-butyl 2-(6-acetamido-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetate (168e) (0.77 g, 2.167 mmol) in DCM (15 mL) using TFA (2.470 g, 21.67 mmol) and stirring at RT for 16 h.

314

This gave after work up TFA sait of 2-(6-acetamido-4-amino-9H-pyrimido[4,5-b]indol-9yl)acetic acid (168f) (1.34 g) as a pale-yellow solid; ’H NMR (300 MHz, DMSO-dô) δ 10.04 (s, 1H), 8.66 (s, 2H), 8.62 (s, 1H), 8.54 (d, J = 1.9 Hz, 1H), 7.76 (d, J = 8.8 Hz, 1H), 7.54 (dd, J = 8.8,1.9 Hz, 1H), 5.27 (s, 2H), 2.09 (s, 3H). ¹⁹F NMR (282 MHz, DMSO- ifc) δ -74.68;

MS (ES+): 300 (M+l); (ES-): 298 (M-l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(6-acetamido-4-amino-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo [3.1.0]hexane-3-carboxamide (168g)

Compound 168g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(6-acetamido-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (168f) (200 mg, 0.484 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (170 mg, 0.532 mmol), HATU (276 mg, 0.726 mmol), DIPEA (313 mg, 2.420 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 to 7%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-acetamido-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (168g) (70 mg, 26% yield) HCl sait as a white solid; ²H NMR (300 MHz, DMSO-de) δ 10.77 (s, 1H, D2O exchangeable), 10.09 (s, 1H, D₂O exchangeable), 8.77 (s, 2H, D₂O exchangeable), 8.66 (s,

1H), 8.52 (d, J = 1.9 Hz, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.75 - 7.60 (m, 2H), 7.55 (dd, J = 8.8,

1.8 Hz, 1H), 7.31 (d, 1H), 5.76 (d, J = 17.4 Hz, 1H), 5.40 (d, J = 17.3 Hz, 1H), 4.42 (dd, J = 9.0, 5.4 Hz, 1H), 3.94 - 3.86 (m, 1H), 2.40 - 2.26 (m, 1H), 2.26 - 2.15 (m, 1H), 2.08 (s, 3H), 1.97 - 1.85 (m, 1H), 1.12 - 0.98 (m, 1H), 0.83 - 0.72 (m, 1H); MS (ES+): 563/565 (M+l), (ES-): 561/563 (M-l); Analysis calculated for C25H23BrN8O3.L75HCl.3H2O: C, 44.08; H, 25 4.55; N, 16.45; Found: C, 44.12; H, 4.67; N, 15.87.

Scheme 169

315

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-N-(cyclopropylmethyl)-9H-pyrimido[4,5-b]indole6-carboxamide (169f)

Step-1: Préparation of 4-amino-9H-pyrimido[4,5-b]indole-6-carboxylic acid (169a) Compound 169a was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-9H-pyrimido[4,5-b]indole-6-carboxylate (39c) (400 mg, 1.651 mmol) in THF (5 ml.) and MeOH (5 mL) using a solution of lithium hydroxide hydrate (424 mg, 9.91 mmol) in water (5 mL) and stirring at RT for 16 h. Additional lithium hydroxide hydrate (212 mg) in water (2 mL) and stirring for 6 h at RT was required for complété hydrolysis.

This gave after work up 4-amino-9H-pyrimido[4,5-b]indole-6-carboxyhc acid (169a) (305 mg, 81% yield) as a gray solid which was used as such for the next step; Y NMR (300 MHz, DMSO-dô) δ 12.16 (s, 1H), 8.90 (d, J = 1.5 Hz, 1H), 8.27 (s, 1H), 7.97 (dd, J = 8.5,1.5 Hz, 1H), 7.48 (d, J = 8.5 Hz, 1H), 7.32 (s, 2H); MS (ES+): 229.00 (M+l).

Step-2: Préparation of 4-amino-N-(cyclopropylmethyl)-9H-pyrimido[4,5-b]indole-6carboxamide (169c)

Compound 169c was prepared according to the procedure reported in scheme-119, from 4amino-9H-pyrimido[4,5-b]indole-6-carboxylic acid (169a) (100 mg, 0.438 mmol) in DMF (3 ml,) using cyclopropylmethanamine (169b) (93 mg, 1.315 mmol), HOBT (29.6 mg, 0.219 mmol), EDC (168 mg, 0.876 mmol), DIPEA (0.153 mL, 0.876 mmol) and stirring at RT for 41 h. This gave after workup 4-amino-N-(cyclopropylmethyl)-9H-pyrimido[4,5-b]mdole-6

316 carboxamide (169c) (12 mg, 10% yield) as a yellow solid which was used as such for the next step; MS (ES+): 282.10 (M+l).

Step-3: Préparation of tert-butyl 2-(4-amino-6-((cyclopropylmethyl)carbamoyl)-9Hpyrimido[4,5-b]indol-9-yl)acetate (169d)

Compound 169d was prepared according to the procedure reported in step-1 of scheme-1, from 4-amino-N-(cyclopropylmethyl)-9H-pyrimido[4,5-b]indole-6-carboxamide (169c) (12 mg, 0.043 mmol) in DMF (4 mL) using tert-butyl 2-bromoacetate (7.56 pL, 0.051 mmol), CS2CO3 (34.7 mg, 0.107 mmol) and stirring at RT for 19 h. This gave after workup tert-butyl 2-(4-amino-6-((cyclopropylmethyl)carbamoyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (169d) (21 mg) as a yellow gum which was used as such for the next step; MS (ES+): 396.20 (M+l).

Step-4: Préparation of 2-(4-amino-6-((cyclopropylmethyl)carbamoyl)-9H-pynmido[4,5b]indol-9-yl)acetic acid (169e)

Compound 169e was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-6-((cyclopropylmethyl)carbamoyl)-9H-pynmido[4,5-b]mdol-915 yl)acetate (169d) (0.017 g, 0.043 mmol) in DCM (5 mL) using TFA and stirring at RT. This gave after workup 2-(4-amino-6-((cyclopropylmethyl)carbamoyl)-9H-pyrimido[4,5-b]indol9-yl)acetic acid (169e) (15 mg) and was used as such for the next step; MS (ES+): 340.20 (M+l).

Step-5; Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-220 azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-N-(cyclopropylmethyl)-9H-pyrimido[4,5-b]indole6-carboxamide (169f).

Compound 169f was prepared according to the procedure reported m step-3 of scheme-1, from TFA sait of 2-(4-amino-6-((cyclopropylmethyl)carbamoyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (169e) (14.59 mg, 0.043 mmol) in DMF (7 mL) using HCl sait of (1R,3S,5R)25 N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (55 mg, 0.172 mmol), HATU (65 mg, 0.172 mmol), DIPEA (0.045 mL, 0.258 mmol) and stirring at RT for 18 h. This gave after workup and purification by flash column chromatography [silica gel (4 g), eluting with dichloromethane/methanol (1:0 to 19:1)] followed by conversion of product obtained to HCl sait by dissolving in acetonitrile (2 mL) and 0.1% aq. HCl (15 mL) and lyophilization 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-N-(cyclopropylmethyl)-9H-pyrimido[4,5-b]indole21436

317

6-carboxamide (169f) (4 mg, 15% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-d₆) δ 10.76 (s, 1H), 8.97 (s, 1H), 8.58 - 8.44 (m, 2H), 8.06 - 7.97 (m, 2H), 7.75 7.64 (m, 2H), 7.31 (d, J = 7.7 Hz, 1H), 5.74 (d, J = 17.4 Hz, 1H), 5.41 (d, J = 17.4 Hz, 1H), 4.48 - 4.35 (m, 1H), 3.95 - 3.86 (m, 1H), 3.31 - 3.10 (m, 2H), 2.41 - 2.12 (m, 2H), 1.97 -

1.83 (m, 1H), 1.30 - 1.01 (m, 2H), 0.84 - 0.70 (m, 1H), 0.54 - 0.39 (m, 2H), 0.36 - 0.21 (m,

2H); MS (ES+): 603.15 & 605.10 (M+l); MS (ES-): 601.10 & 603.10 (M-l).

Scheme 170

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(l-hydroxyethyl)-9H-pyrimido[4,5-b]indol-910 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (170c)

Step-1: Préparation of ethyl 2-(4-amino-6-(l-hydroxyethyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (170a)

Compound 170a was prepared according to the procedure reported in scheme-157, from ethyl 2-(6-acetyl-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetate (58a) (32 mg, 0.102 mmol) in

THF (6 mL) and MeOH (3 mL) using sodium borohydride (11.87 mg, 0.307 mmol) and stirring at RT for 24 h. The residue obtained after work up was used as such for the next step; MS (ES+): 301.10 (M+l).

Step-2: Préparation of 2-(4-amino-6-(l-hydroxyethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (170b)

318

Compound 170b was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-6-(l-hydroxyethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (170a) (0.032 mg, 0.102 mmol) in THF (3 mL) and MeOH (3 mL) using a solution of lithium hydroxide hydrate (0.026 g) and stirring at RT for 20 h. The obtained residue after work up 5 was used as such for the next step; MS (ES+): 287.10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(l-hydroxyethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (170c)

Compound 170c was prepared according to the procedure reported in step-3 of scheme-1, 10 from TFA sait of 2-(4-amino-6-(l-hydroxyethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (170b) (29.2 mg, 0.102 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (65 mg, 0.204 mmol), HATU (97 mg, 0.255 mmol), DIPEA (0.071 mL, 0.408 mmol) and stirring at RT for 20 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), 15 eluting with dichloromethane/methanol (1:0 to 19:1)] followed by conversion of product obtained to HCl sait by dissolving in acetonitrile (2 mL) and 0.1% aq. HCl (15 mL) and lyophilization(lR,3S,5R)-2-(2-(4-amino-6-(l-hydroxyethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (170c) (12 mg, 21% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.76 (s, 1H), 20 8.62 (s, 2H), 8.61 (s, 1H), 8.45 (s, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.67

- 7.55 (m, 2H), 7.31 (d, J = 7.7 Hz, 1H), 5.75 (d, J = 17.4 Hz, 1H), 5.41 (d, J = 17.3 Hz, 1H), 4.91 (q, J = 6.4 Hz, 1H), 4.40 (dd, J = 9.1, 5.5 Hz, 1H), 3.99 - 3.86 (m, 1H), 2.43 - 2.10 (m, 2H), 2.01 - 1.78 (m, 1H), 1.43 (d, J = 6.4 Hz, 3H), 1.14 - 0.99 (m, 1H), 0.82 - 0.69 (m, 1H); MS (ES+): 550.10 & 552.10 (M+l); MS (ES-): 548.00 & 550.10 (M-l).

Scheme 171

319

Préparation of (S)-3-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)thiazolidine-2-carboxamide (171b)

Compound 171b was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (75 mg, 0.177 mmol) in DMF (2 mL) using TFA sait of (S)-N-(6-bromopyridin-2yl)thiazolidine-2-carboxamide (171a) (71.1 mg, 0.177 mmol; prepared according to the procedure reported by Yang, Chao-Yie in ACS Med. Chem. Lett. 2016, 7,1092-1096.), HATU (101 mg, 0.265 mmol), DIPEA (0.154 mL, 0.884 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (S)-3-(2-(4-amino-6-(trifhioromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)thiazolidine-2-carboxamide (171b) (61 mg, 60% yield) HCl sait as a white solid; ^XH NMR (300 MHz, DMSO-î/₆) (a mixture of two rotamers) δ 11.39 and 11.03 (2s, 1H, D₂O exchangeable), 8.99 (s, 1H), 8.62 (m, 3H, 2H D₂O exchangeable), 8.11 and 7.96 (2dd, J = 8.4, 4.5 Hz, 2H), 7.87 (dd, J = 8.8,1.7 Hz, 1H), 7.73 (t, J = 8.0 Hz, 1H), 7.42 and 7.34 (2d, J = 7.7 Hz, 1H), 5.75 - 5.46 (m, 3H), 4.40 (dt, J = 9.9, 4.8 Hz, 1H), 4.21 - 4.05 (m, 1H), 3.38 (dd, J = 8.3, 5.4 Hz, 2H). ¹⁹F NMR (282 MHz, DMSO-efe) δ -58.73. MS (ES+): 580.0 (M+l); (ES-): 578.0 (M-l); Analysis calculated for C₂₂Hi7BrF₃N7O₂S

1.75H₂O.HC1: C, 40.75; H, 3.34; Cl, 5.47; N, 15.12; Found: C, 40.84; H, 3.18; Cl, 5.40; N,

14.93.

Scheme 172

57f 172a

Préparation of (lR,3S,5R)-2-(2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)25 N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide(172a)

320

Compound 172a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (57f) (50 mg, 0.129 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (43.1 mg, 0.129 mmol), HATU (73.8 mg, 0.194 mmol), DIPEA (0.113 mL, 0.647 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(610 bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (172a) (51mg, 72% yield) HCl sait as a white solid; ¹H NMR (300 MHz, DMSO-î/ô) δ 10.74 (s, 1H, D₂O exchangeable), 8.65 (s, 2H, D₂O exchangeable), 8.60 (s, 1H), 8.42 (d, J = 8.8 Hz, 1H), 8.02 (d, J = 8.1 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.36 - 7.25 (m, 2H), 7.06 (dd, J = 8.8, 2.3 Hz, 1H), 5.69 (d, J = 17.3 Hz, 1H), 5.38 (d, J = 17.3 Hz, 1H), 4.38 (dd, J = 9.1, 6.0 Hz, 1H), 3.89 (s, 3H), 3.69 (dd, J = 5.6, 2.4 Hz, 1H), 2.47 (d, J = 9.0 Hz, 1H), 2.00 (dd, J = 13.3, 6.0 Hz,

1H), 1.32 (s, 3H), 1.03 (t, J = 5.4 Hz, 1H), 0.92 (dd, J = 5.3, 2.4 Hz, 1H); MS (ES+): 550.1 (M+l), 572.1 (M+Na); (ES-): 548.1 (M-l); Analysis calculated for

C₂5H₂4BrN₇O₃.1.2HC1.1.5H₂O: C, 48.34; H, 4.58; Cl, 6.85; N, 15.78; Found: C, 48.20; H, 4.62; Cl, 6.50; N, 15.83.

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (173a)

Compound 173a was prepared according to the procedure reported in step-3 of scheme-1, 25 from TFA sait of 2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (54f) (50 mg, 0.129 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5

321 methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (43.1 mg, 0.129 mmol), HATU (73.8 mg, 0.194 mmol), DIPEA (0.113 mL, 0.647 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (173a) (47 mg, 66% yield) HCl sait as a white solid; ¹H NMR (300 MHz, DMSO-î/ô) δ 10.73 (s, 1H, D2O exchangeable), 8.69 — 8.50 (m, 3H, 2H D2O exchangeable), 8.05 (dd, J = 16.9, 8.1 Hz, 2H),

7.71 (t, J = 8.0 Hz, 1H), 7.40 - 7.27 (m, 2H), 7.14 (d, J = 8.1 Hz, 1H), 5.76 (d, J = 17.0 Hz,

1H), 5.55 (d, J = 16.9 Hz, 1H), 4.39 (dd, J = 9.0, 5.9 Hz, 1H), 3.93 (s, 3H), 3.62 (dd, J = 5.4, 2.3 Hz, 1H), 2.49 - 2.41 (m, 1H), 2.05 - 1.91 (m, 1H), 1.31 (s, 3H), 1.06 (t, J = 5.4 Hz, 1H), 0.83 (dd, J = 5.3, 2.3 Hz, 1H); MS (ES+): 550.1 (M+l), 572.1 (M+Na); (ES-): 548.1 (M-l); Analysis calculated for C25H24BrN7O3.HCl.2H2O: C, 48.20; H, 4.69; Cl, 5.69, N, 15.74,

Found: C, 48.00; H, 4.59; Cl, 5.46; N, 15.60 .

Scheme 174

322

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo [3.1.0]hexane-3-carboxamide (174g)

Step-1: Préparation of N-(2-bromo-6-(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (174b)

Compound 174b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-6-(trifluoromethyl)aniline (174a) (5 g, 20.83 mmol; CAS # 58458-13-2) in DCM (50 mL) using triethylamine (4.94 mL, 35.4 mmol) and trifluoroacetic acid anhydride (4.34 mL, 31.2 mmol) and stirring at RT for 16 h. This gave after workup N-(2-bromo-6(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (174b) as a pale yellow solid (7 g, 100% 10 yield) which was used as such for next step; ¹H NMR (300 MHz, DMSO-î/ô) δ 11.72 (s, 1H),

8.16 (dd, J = 8.2,1.3 Hz, 1H), 7.91 (dd, J = 8.0,1.3 Hz, 1H), 7.61 (t, J = 8.0 Hz, 1H); MS (ES-): 333.90 (M-l).

Step-2: Préparation of 2-amino-7-(trifluoromethyl)-lH-indole-3-carbonitrile (174c)

Compound 174c was prepared according to the procedure reported in step-1 of scheme-11, 15 from N-(2-bromo-6-(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (174b) (3 g, 8.93 mmol) in DMSO (9 mL) using malononitrile (0.675 mL, 10.71 mmol), L-proline (0.206 g, 1.786 mmol), Cul (0.170 g, 0.893 mmol), a solution of K2CO3 (2.468 g, 17.86 mmol) in water (9 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [SiO₂gel (80 g), eluting with EtOAc in hexane from 0-50%] 220 amino-7-(trifluoromethyl)-lH-indole-3-carbonitrile (174c) (650 mg, 32% yield) as a yellow solid; Ή NMR (300 MHz, DMSO-rf₆) δ 11.04 (s, 1H), 7.43 (d, J = 7.5 Hz, 1H), 7.29 - 7.09 (m, 2H), 6.79 (s, 2H); MS (ES-): 224.00 (M-l).

Step-3: Préparation of 8-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-4-amine (174d)

Compound 174d was prepared according to the procedure reported in step-1 of scheme-6, 25 from 2-amino-7-(trifluoromethyl)-lH-indole-3-carbonitrile (174c) (600 mg, 2.66 mmol) using trimethyl orthoformate (5830 pL, 53.3 mmol), AcOH (762 pL, 13.32 mmol), NH4OAC (1027 mg, 13.32 mmol) and heating at 100 °C for 16 h in a pressure vessel. This gave after workup acetic acid sait of 8-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-4-amine (174d) (478 mg, 58% yield) as a yellow solid; ^]H NMR (300 MHz, DMSO-îL) δ 12.30 (s, 1H), 11.98 (s, 30 1H), 8.61 (d, J = 7.9 Hz, 1H), 8.35 (s, 1H), 7.67 (d, J = 7.7 Hz, 1H), 7.50 - 7.29 (m, 3H),

1.91 (s, 3H); MS (ES+): 253.00 (M+l).

323

Step-4: Préparation of ieri-butyl 2-(4-amino-8-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (174e)

Compound 174e was prepared according to the procedure reported in step-1 of scheme-1, from acetic acid sait of 8-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-4-amine (174d) (450 mg, 1.441 mmol) in DMF (10 mL) using ieri-butyl 2-bromoacetate (0.234 mL, 1.585 mmol), CS2CO3 (1033 mg, 3.17 mmol) and stirring at RT for 15 h. This gave after work up and purification using flash column chromatography [SiO₂ gel (40 g), eluting with ethyl acetate in hexanes from 0-100%] tert-butyl 2-(4-amino-8-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (174e) (180 mg, 34% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-cfc) δ 8.56 - 8.48 (m, 2H), 7.54 (d, J = 7.6 Hz, 1H), 7.20 (t, J = 7.7 Hz, 1H), 5.14 (s, 2H), 1.42 (s, 9H); MS (ES+): 367.10 (M+l).

Step-5: Préparation of 2-(4-amino-8-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (174Ç

Compound 174f was prepared according to the procedure reported in step-2 of scheme-1, from teri-butyl 2-(4-amino-8-(trifhioromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (174e) (175 mg, 0.478 mmol) using 20% TFA in DCM (2742 μΕ, 7.17 mmol) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-8-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (174f) (198 mg, 98% yield) as a pale yellow solid; MS (ES+): 311.08 (M+l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (174g)

Compound 174g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (174f) (60mg, 0.141 mmol) in DMF (1.5 mL) using TFA sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (56.0 mg, 0.141 mmol), HATU (81 mg, 0.212 mmol), DIPEA (0.123 mL, 0.707 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-(trifluoromethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (174g) (75

324 mg, 92% yield) HCl sait as a white solid; ^XH NMR (300 MHz, DMSO-t/₆/D₂O) δ 8.82 - 8.69 _(m ’ 2H), 7.98 (d, J = 8.1 Hz, 1H), 7.88 (d, J = 7.7 Hz, 1H), 7.69 (t, J = 8.0 Hz, 1H), 7.60 (t, J = 7.8 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 6.01 (d, J = 17.2 Hz, 1H), 5.64 (d, J = 17.2 Hz, 1H), 4.45 (dd, J = 9.0, 5.7 Hz, 1H), 3.73 (ddd, J = 7.4, 5.5, 2.3 Hz, 1H), 2.36 (dd, J = 13.2, 9.5 Hz, 5 1H), 2.23 (dt, J = 13.0, 6.2 Hz, 1H), 1.96 (tt, J = 6.6, 3.5 Hz, 1H), 1.10 (dt, J = 8.6, 5.5 Hz,

1H)’, 0.86 (td, J = 5.3, 2.3 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-d₆) δ -59.64. MS (ES+): 574.00 & 576.1 (M+l); (ES-): 572.10 & 574.10 (M-l); Analysis calculated for C24Hi₉BrF₃N7O2 1.5H₂O.HC1: C, 45.19; H, 3.63; Cl, 5.56; N, 15.37; Found: C, 44.94; H, 3.71; Cl, 5.32; N, 15.36.

Scheme 175

Préparation of(lR,3S,5R)-2-(2-(4-amino-8-(trifluoromethyl)-9H-pyrimido[4,5-b]mdol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carbo_Xamide (175a)

Compound 175a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (174f) (60 mg, 0.141 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6bromop_yridin-2-yl)-5-methyl-2-azabic_yclo[3.1.0]hexane-3-carboxamide (8a) (47 mg, 0.141 mmol), HATU (81 mg, 0.212 mmol) DIPEA (0.123 mL, 0.707 mmol) and stirring at RT for

16 h. This gave after workup and purification by flash column chromatography [silica gel (12

g), using DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (contammg 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-(trifluoromethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamrde (175a) (47 mg, 57% yield) HCl sait as a white solid; ^XH NMR (300 MHz, DMSO-J₆/D₂O) δ

8.78 - 8.57 (m, 2H), 7.94 (d, J = 8.2 Hz, 1H), 7.79 (d, J = 7.7 Hz, 1H), 7.64 (t, J = 8.0 Hz,

325

1H), 7.58 - 7.40 (m, 1H), 7.26 (d, J = 7.7 Hz, 1H), 5.85 (d, J = 17.2 Hz, 1H), 5.52 (d, J =

17.0 Hz, 1H), 4.36 (dd, J = 9.0, 6.1 Hz, 1H), 3.47 - 3.42 (m, 1H), 2.43 - 2.38 (m, 1H), 1.95 (dd, J = 13.3, 6.0 Hz, 1H), 1.26 (s, 3H), 1.02 - 0.92 (m, 2H); ¹⁹F NMR (282 MHz, DMSO-ifc) δ -59.60; MS (ES+): 588.1 (M+l), 610.1 (M+Na); (ES-): 586.1 (M-l); Analysis calculated for C25H2iBrF₃N7O₂ 2H₂O.HC1: C, 45.44; H, 3.97; Cl, 5.36; N, 14.84; Found: C, 45.71; H, 3.97; Cl, 5.09; N, 14.63.

Scheme 176

Préparation of (lR,3S,5R)-2-(2-(4-amino-5-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N10 (6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (176a)

Compound 176a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-5-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (56f) (50mg, 0.135 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (44.9 mg, 0.135 mmol), HATU 15 (77 mg, 0.203 mmol), DIPEA (0.118 mL, 0.675 mmol) and stirring at RT for 4 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%](lR,3S,5R)-2-(2-(4-amino-5-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(620 bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (176a) (53 mg, 73% yield) HCl sait as a white solid; ^JH NMR (300 MHz, DMSO-îL) δ 10.77 (s, 1H, D2O exchangeable), 8.63 (s, 1H), 8.01 (d, J = 8.1 Hz, 1H), 7.86 (s, 2H, D₂O exchangeable), 7.70 (t, J = 8.0 Hz, 1H), 7.46 (dt, J = 15.3, 8.0 Hz, 2H), 7.32 (d, J = 7.7 Hz, 1H), 7.18 (d, J = 7.0 Hz, 1H), 5.71 (d, J = 17.4 Hz, 1H), 5.37 (d, J = 17.3 Hz, 1H), 4.37 (dd, J = 9.1, 5.9 Hz, 1H), 25 3.70 (dd, J = 5.5, 2.4 Hz, 1H), 2.96 (s, 3H), 2.49 - 2.41 (m, 1H), 1.98 (dd, J = 13.2, 5.9 Hz,

326

1H), 1.31 (s, 3H), 1.02 (t, J = 5.4 Hz, 1H), 0.93 (dd, J = 5.4, 2.4 Hz, 1H). MS (ES+):

534.1(M+1), 556.1 (M+Na); (ES-): 532.0 (M-l); Analysis calculated for C25H₂4BrN?O₂ 1.25H₂O.HC1: C, 50.60; H, 4.67; Cl, 5.97; N, 16.52; Found: C, 50.63; H, 4.76; Cl, 5.73; N, 16.55.

Scheme 177

O

II a^ci o

177b

CF₃

177d

Préparation of (2S,4S)-l-(2-(4-amino-6-(trifhioromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-4-cyanopyrrolidine-2-carboxamide (177d)

Step-1: Préparation of (2S,4S)-teri-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-410 cyanopyrrolidine-l-carboxylate (177b)

Compound 177b was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4S)-l-(tert-butoxycarbonyl)-4-cyanopyrrolidine-2-carboxylic acid (177a) (0.5 g, 2.081 mmol; CAS # 132622-71-0) in DCM (15 mL) using 1-methyl-lH-imidazole (0.415 mL, 5.20 mmol), methanesulfonyl chloride (0.193 mL, 2.497 mmol), 6-bromopyridin-215 amine (0.360 g, 2.081 mmol) and stirring at RT for 18 h. This gave after workup (2S,4S)-ierZbutyl 2-((6-bromopyridin-2-yl)carbamoyl)-4-cyanopyrrolidine-l-carboxylate (177b) (0.5 g, 61% yield); ^XH NMR (300 MHz, DMSO-dô) δ 11.05 (d, J = 14.5 Hz, 1H), 8.17 - 8.05 (m, 1H), 7.77 (q, J = 7.7 Hz, 1H), 7.37 (d, J = 7.7 Hz, 1H), 4.39 (q, J = 7.9 Hz, 1H), 3.82 (d, J =

327

13.5 Hz, 1H), 3.49 (d, J = 6.0 Hz, 2H), 2.70 (dd, J = 14.3, 7.6 Hz, 1H), 2.10 (dt, J = 13.7, 8.0 Hz, 1H), 1.33 (d, J = 43.3 Hz, 9H); MS (ES+): 395.10 (M+l).

Step-2: Préparation of (2S,4S)-N-(6-bromopyridin-2-yl)-4-cyanopyrrolidine-2-carboxamide (177c)

Compound 177c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4S)-tert-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-4-cyanopyrrolidine-lcarboxylate (177b) (200 mg, 0.506 mmol) in DCM (2.5 mL) using TFA (404 mg, 3.54 mmol) and stirring overnight at RT. This gave after workup TFA sait of (2S,4S)-N-(6bromopyridin-2-yl)-4-cyanopyrrolidine-2-carboxamide (177c) (0.2 g, 97% yield) as a clear gel; Ή NMR (300 MHz, DMSO-d₆) δ 11.52 (s, 1H), 8.06 (d, J = 8.1 Hz, 1H), 7.83 (t, J = 8.0 Hz, 1H), 7.46 (d, J = Ί.Ί Hz, 1H), 4.48 (t, J = 8.3 Hz, 1H), 3.67 (dt, J = 12.9, 8.0 Hz, 2H), 3.55 (dd, J = 9.9, 5.8 Hz, 1H), 2.87 (dt, J = 13.6, 7.9 Hz, 1H), 2.32 (dt, J = 14.3, 7.3 Hz, 1H); MS (ES+): 295.00 (M+l).

Step-3: Préparation of (2S,4S)-l-(2-(4-amino-6-(trifhioromethyl)-9H-pyrimido[4,5-b]indol-915 yl)acetyl)-N-(6-bromopyridin-2-yl)-4-cyanopyrrolidine-2-carboxamide (177d)

Compound 177d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (2S,4S)-N-(6-bromopyridin-2-yl)-4-cyanopyrrolidine-2-carboxamide (177c) (0.2 g, 0.489 mmol) in DMF (1.5 mL) using TFA sait of 2-(4-amino-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (lie) (0.207 g, 0.489 mmol), HATU (0.279 g, 0.733 20 mmol), DIPEA (0.426 mL, 2.444 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(625 bromopyridin-2-yl)-4-cyanopyrrolidine-2-carboxamide (177d) (173 mg, 60% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) (a mixture of two rotamers) δ 11.40 and 10.97 (2s, 1H, D2O exchangeable), 8.93 (s, 1H), 8.54 (s, 1H), 8.33 (s, 2H, D2O exchangeable), 8.17 and 8.01 (2d, J = 8.2 Hz, 1H), 7.84 (d, J = 4.7 Hz, 2H), 7.71 (t, J = 7.9 Hz, 1H), 7.43 and 7.34 (2d, J - 7.7 Hz, 1H), 5.77 and 5.50 (2s, 2H), 4.55 (t, J = 7.5 Hz, 1H), 30 4.39 (dd, J = 10.0, 7.8 Hz, 1H), 4.07 (dd, J = 10.1, 7.8 Hz, 1H), 3.77 - 3.68 (m, 1H), 2.82 2.68 (m, 1H), 2.19 (dt, J = 14.0, 7.6 Hz, 1H). ¹⁹F NMR (282 MHz, DMSO-cfe) δ -58.53. MS (ES+): 587.1 (M+l); (ES-): 585.0 (M-l); Analysis calculated for C24Hi8BrF3NsO2 2H2O.HCI: C, 43.69; H, 3.51; Cl, 5.37; N, 16.98; Found: C, 43.70; H, 3.49; Cl, 4.98; N, 16.62.

328

Scheme 178

178d

Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-4-cyanopyrrolidine-2-carboxamide (178d)

Step-1: Préparation of (2S,4R)-tert-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-4cyanopyrrolidine-l-carboxylate (178b)

Compound 178b was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4R)-l-(terLbutoxycarbonyl)-4-cyanopyrrolidine-2-carboxylic acid (178a) (0.5 g, 2.081 mmol; CAS # 273221-94-6) in DCM (15 mL) using 1-methyl-lH-imidazole (0.415 mL, 5.20 mmol), methanesulfonyl chloride (0.193 mL, 2.497 mmol), 6-bromopyridin-2amine (0.360 g, 2.081 mmol) and stirring at RT for 18 h. This gave after workup (2S,4R)tert-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-4-cyanopyrrolidine-l-carboxylate (178b) (747 mg, 91% yield); Ή NMR (300 MHz, DMSO-î/₆) δ 11.07 (d, J = 17.6 Hz, 1H), 8.07 (dd, J = 12.9, 8.1 Hz, 1H), 7.77 (q, J = 8.0 Hz, 1H), 7.37 (d, J = 7.7 Hz, 1H), 4.53 (ddd, J = 13.0, 8.2,

4.9 Hz, 1H), 3.73 (dd, J = 10.4, 7.1 Hz, 1H), 3.57 (dt, J = 10.6, 5.7 Hz, 1H), 3.46 (tt, J = 7.1,

4.2 Hz, 1H), 2.46 (d, J = 8.2 Hz, 1H), 2.30 (dt, J = 12.6, 6.1 Hz, 1H), 1.40 (s, 3H), 1.27 (s, 6H).

Step-2: Préparation of (2S,4R)-N-(6-bromopyridin-2-yl)-4-cyanopyrrohdine-2-carboxamide (178c)

329

Compound 178c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-iert-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-4-cyanopyrrolidine-lcarboxylate (178b) (0.2 g, 0.506 mmol) in DCM (2.5 mL) using TFA (0.273 mL, 3.54 mmol) and stirring overnight at RT. This gave after workup TFA sait of (2S,4R)-N-(6-bromopyridin5 2-yl)-4-cyanopyrrolidine-2-carboxamide (178c) (0.2 g, 97% yield) as a clear gel; ¹H NMR (300 MHz, DMSO-dô) 8 11.50 (s, 1H), 8.04 (d, J = 8.2 Hz, 1H), 7.82 (t, J = 8.0 Hz, 1H), 7.45 (dd, J = 7.8, 0.7 Hz, 1H), 4.62 (t, J = 7.8 Hz, 1H), 3.76 - 3.51 (m, 3H), 2.68 (ddd, J = 11.2, 8.0, 5.5 Hz, 1H), 2.49 - 2.40 (m, 1H); MS (ES+): 295.00 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-6-(trifhioromethyl)-9H-pyrimido[4,5-b]indol10 9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-cyanopyrrolidine-2-carboxamide (178d)

Compound 178d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (2S,4R)-N-(6-bromopyridin-2-yl)-4-cyanopyrrolidine-2-carboxamide (178c) (200 mg, 0.488 mmol) in DMF (5 mL) using TFA sait of 2-(4-amino-6(trifhioromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (207 mg, 0.488 mmol), 15 HATU (0.278 g, 0.732 mmol), DIPEA (0.425 mL, 2.440 mmol) and stirring at RT for 16 h.

This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%](2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-920 yl)acetyl)-N-(6-bromopyridin-2-yl)-4-cyanopyrrolidine-2-carboxamide (178d) (177 mg, 62% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-Jô) (a mixture of two rotamers) δ 11.38 and 11.00 (2s, 1H, D₂O exchangeable), 8.95 (s, 1H), 8.56 and 8.52 (2s, 1H), 8.42 (s, 2H, D₂O exchangeable), 8.16 and 7.99 (2d, J = 8.2 Hz, 1H), 7.85 (d, J = 2.2 Hz, 2H), 7.71 (t, J = 8.0 Hz, 1H), 7.43 and 7.33 (2d, J = Ί.Ί Hz, 1H), 5.53 (d, J = 2.91 Hz , 2H), 4.68 (dd, J =

8.4, 4.5 Hz, 1H), 4.31 (dd, J = 10.0, 7.5 Hz, 1H), 4.19 (dd, J = 10.0, 6.9 Hz, 1H), 3.70 (p, J =

7.2 Hz, 1H), 2.59 - 2.52 (m, 1H), 2.46 - 2.36 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-d₆) δ 58.55; MS (ES+): 587.1 (M+l); (ES-): 585.0 (M-l).

Scheme 179

Préparation of (S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)indoline-2-carboxamide (179d)

Step-1: Préparation of (S)-tert-butyl 2-((6-bromopyridin-2-yl)carbamoyl)indoline-l5 carboxylate (179b)

Compound 179b was prepared according to the procedure reported in step-1 of scheme-53, from (S)-l-(ieri-butoxycarbonyl)indoline-2-carboxylic acid (179a) (1 g, 3.80 mmol; CAS # 144069-67-0) in DCM (30 mL) using 1-methyl-lH-imidazole (0.780 mL, 9.50 mmol), methanesulfonyl chloride (0.353 mL, 4.56 mmol), 6-bromopyridin-2-amine (0.657 g, 3.80 mmol) and stirring at RT for 18 h. This gave after workup (S)-tert-butyl 2-((6-bromopyridm2-yl)carbamoyl)indoline-l-carboxylate (179b) (1.49 g, 94% yield); ^XH NMR (300 MHz, DMSO-dô) δ 11.21 (s, 1H), 8.08 (d, J = 8.3 Hz, 1H), 7.77 (t, J = 8.3 Hz, 2H), 7.37 (d, J = 7.7 Hz, 1H), 7.17 (d, J = 7.4 Hz, 2H), 6.94 (t, J = 7.4 Hz, 1H), 5.04 (dd, J = 11.1, 4.6 Hz, 1H), 3.49 (dd, J = 17.0,11.2 Hz, 1H), 3.06 (d, J = 17.0 Hz, 1H), 1.44 (d, J = 53.8 Hz, 9H); MS (ES+): 418.10 (M+l).

Step-2: Préparation of (S)-N-(6-bromopyridin-2-yl)indoline-2-carboxamide (179c)

Compound 179c was prepared according to the procedure reported in step-2 of scheme-1, from (S)-ieri-butyl 2-((6-bromopyridin-2-yl)carbamoyl)indoline-l-carboxylate (179b) (1 g, 2.391 mmol) in DCM (10 mL) using TFA (1.289 mL, 16.74 mmol) and stirring overnight at 20 RT. This gave after work up and purification using reverse phase column chromatography

331

[C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (S)-N(6-bromopyridin-2-yl)indoline-2-carboxamide (179c) (0.66 g, 78% yield) HCl sait as an off white solid; Ή NMR (300 MHz, DMSO-dô) δ 10.59 (s, 1H), 8.10 (d, J = 8.1 Hz, 1H), 7.76 (t, J = 8.0 Hz, 1H), 7.37 (d, J = 7.7 Hz, 1H), 7.06 - 6.92 (m, 2H), 6.68 - 6.56 (m, 2H), 4.51 (dd, 5 J = 10.5, 6.9 Hz, 1H), 3.35 (dd, J = 16.2,10.5 Hz, 1H), 3.11 (dd, J = 16.2, 6.8 Hz, 1H); MS (ES+): 318.00 (M+l).

Step-3: Préparation of (S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)indoline-2-carboxamide (179d)

Compound 179d was prepared according to the procedure reported in step-3 of scheme-1, 10 from HCl sait of (S)-N-(6-bromopyridin-2-yl)indoline-2-carboxamide (179c) (83mg, 0.234 mmol) in DMF (2 mL) using TFA sait of 2-(4-amino-6-(trifhioromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (lie) (99 mg, 0.234 mmol), HATU (133 mg, 0.351 mmol), DIPEA (0.204 mL, 1.170 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 m DCM from 0 to

50%] followed by purification using reverse phase column chromatography [C18 column (50

g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (S)-l-(2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)indoline-2carboxamide (179d) (71 mg, 49% yield) HCl sait as a white solid; ^]H NMR (300 MHz, DMSO-dô) (a mixture of two rotamers) δ 11.58 and 11.08 (2s, 1H, D₂O exchangeable), 8.98 (s, 1H), 8.55 (s, 1H), 8.45 (s, 3H, D₂O exchangeable), 8.14 (d, J = 8.2 Hz, 1H), 8.01 - 7.70 (m, 4H), 7.44 - 7.25 (m, 2H), 7.16 (t, J = 7.6 Hz, 1H), 7.05 (t, J = ΊΑ Hz, 1H), 5.80 - 5.60 (m, 2H), 5.16 (d, J = 17.1 Hz, 1H), 3.82 and 3.58 (2dd, J = 17.1,11.5 Hz, 1H), 3.38 and 3.13 (2d, J = 17.1 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-d₆) δ -58.46; MS (ES+): 610.50 (M+l); (ES-): 608.50 (M-l).

Scheme 180

332

Préparation of (2S,4S)-4-amino-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (180e)

Step-1: Préparation of (2S,4S)-tert-butyl 4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-25 ((6-bromopyridin-2-yl)carbamoyl)pyrrolidine-l-carboxylate (180b)

Compound 180b was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-l-(ieributoxycarbonyl)pyrrolidine-2-carboxylic acid (180a) (1 g, 2.210 mmol; CAS # 174148-03-9) in DCM (30 mL) using 1-methyl-lH-imidazole (0.440 mL, 5.52 mmol), methanesulfonyl chloride (0.205 mL, 2.65 mmol), 6-bromopyridin-2-amine (0.382 g, 2.210 mmol) and stirring at RT for 18 h. This gave after workup (2S,4S)-teri-butyl 4-((((9H-fluoren-9yl)methoxy)carbonyl)amino)-2-((6-bromopyridin-2-yl)carbamoyl)pyrrolidine-l-carboxylate (180b) (1.22 g, 91% yield); Ή NMR (300 MHz, DMSO-d₆) δ 10.98 (d, J = 18.4 Hz, 1H), 8.22 - 8.06 (m, 1H), 7.89 (d, J = 7.5 Hz, 2H), 7.78 (t, J = 8.1 Hz, 1H), 7.67 (d, J = 7.4 Hz,

2H), 7.56 (d, J = 7.7 Hz, 1H), 7.36 (ddt, J = 20.7,12.9, 7.3 Hz, 5H), 4.40 - 4.27 (m, 3H),

4.21 (t, J = 6.8 Hz, 1H), 4.02 (d, J = 8.1 Hz, 1H), 3.82 - 3.56 (m, 1H), 3.14 (s, 1H), 2.51 (p, J = 1.9 Hz, 1H), 1.78 (d, J = 10.0 Hz, 1H), 1.39 (s, 3H), 1.25 (s, 6H); MS (ES+): 607.20 (M+l).

Step-2: Préparation of (9H-fluoren-9-yl)methyl ((3S,5S)-5-((6-bromopyridin-220 yl)carbamoyl)pyrrolidin-3-yl)carbamate (180c)

I,

333

Compound 180c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4S)-tert-butyl 4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((6-bromopyridin2-yl)carbamoyl)pyrrolidine-l-carboxylate (180b) (1 g, 1.646 mmol) in DCM (8 mL) using TFA (0.888 mL, 11.52 mmol) and stirring overnight at RT. This gave after work up and purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (9H-fluoren-9-yl)methyl ((3S,5S)-5-((6bromopyridin-2-yl)carbamoyl)pyrrolidin-3-yl)carbamate (180c) (0.62 g, 69% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-cfc) δ 11.45 (s, 1H), 9.71 (s, 1H), 8.87 (s, 1H), 8.07 (d, J = 8.1 Hz, 1H), 7.93 - 7.79 (m, 3H), 7.67 (d, J = 7.4 Hz, 2H), 7.58 (d, J = 6.2 Hz,

1H), 7.48 - 7.29 (m, 5H), 4.48 - 4.31 (m, 3H), 4.19 (dt, J = 19.5, 6.7 Hz, 2H), 3.40 (s, 1H),

3.16 (d, J = 9.0 Hz, 1H), 2.77 - 2.63 (m, 1H), 1.92 (dt, J = 15.1, 8.1 Hz, 1H); MS (ES+): 507.10 (M+l).

Step-3: Préparation of (9H-fluoren-9-yl)methyl ((3S,5S)-l-(2-(4-amino-6-(trifluoromethyl)9H-pyrimido[4,5-b]indol-9-yl)acetyl)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidm-315 yl)carbamate (180d)

Compound 180d was prepared according to the procedure reported m step-3 of scheme-1, from HCl sait of (9H-fluoren-9-yl)methyl ((3S,5S)-5-((6-bromopyndin-2yl)carbamoyl)pyrrolidin-3-yl)carbamate (180c) (200 mg, 0.368 mmol) in DMF (3 mL) using TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (156 mg, 0.368 mmol), HATU (210 mg, 0.552 mmol), DIPEA (0.320 mL, 1.839 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with DMA-80 in DCM from 0 - 50%] (9H-fluoren9-yl)methyl ((3S,5S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-3-yl)carbamate (180d) (280 mg,

95% yield) as a white solid; MS (ES+): 799.38 (M+l).

Step-4: Préparation of (2S,4S)-4-amino-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (180e) .

To a stirred solution of (9H-fluoren-9-yl)methyl ((3S,5S)-l-(2-(4-amino-6-(trifluoromethyl)9H-pyrimido[4,5-b]indol-9-yl)acetyl)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-330 yl)carbamate (180d) (150mg, 0.188 mmol) in DMF (0.8 mL) was added pipendine (0.185 mL, 1.876 mmol) and stirred at RT for lh. The reaction mixture was concentrated and purified using reverse phase column chromatography [C18 column (50 g), elutmg with ACN

334

in water (containing 0.1% HCl) from 0-100%] yielding (2S,4S)-4-amino-l-(2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)pyrrolidine2-carboxamide (180e) (27 mg, 25% yield) as a white solid; ’H NMR (300 MHz, DMSO-dâ) (a mixture of two rotamers) δ 11.51 and 11.12 (2s, 1H, D2O exchangeable), 8.94 (s, 1H), 8.53 5 (s, 1H), 8.45 - 8.29 and 8.27 - 8.17 (m, 4H, D2O exchangeable), 7.99 (d, J = 8.2 Hz, 1H),

7.90 - 7.78 (m, 2H), 7.72 (t, J = 8.0 Hz, 1H), 7.45 and 7.35 (d, J = 7.7 Hz, 1H), 5.49 (s, 2H), 4.57 (t, J = 7.5 Hz, 1H), 4.23 (dd, J = 10.3, 6.1 Hz, 1H), 4.08 - 3.95 (m, 1H), 3.88 (dd, J = 10.2, 6.2 Hz, 1H), 2.64 (dt, J = 14.3, 7.4 Hz, 1H), 2.05 (dt, J = 13.2, 6.5 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-Jô) δ -58.43. MS (ES+): 576.5 (M+l); 598.5(M+Na); (ES-): 574.6 (M-l);

Analysis calculated for C23H2oBrF3Ns02 2HC1 3.5H2O 0.5DMSO: C, 38.31; H, 4.29; N, 14.89; Found: C, 38.58; H, 4.28; N, 14.59.

Scheme 181

163f 181a

Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[3^,,4':4,5]pyrrolo[2,3-d]pyrimidin-915 yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (181a)

Compound 181a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-9H-pyrido[3',4^,:4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (163f) (96 mg, 0.269 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(620 bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (98 mg, 0.296 mmol), HATU (153 mg, 0.403 mmol), DIPEA (174 mg, 1.344 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 to 7%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1%

HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[3’,4':4,5]pyrrolo[2,3-d]pyrimidin9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide

335 (181a) (62 mg, 44% yield) as a white solid; ^XH NMR (300 MHz, DMSO-dc,) δ 10.78 (s, 1H, D₂O exchangeable), 10.06 (s, 1H), 8.83 (d, J = 6.7 Hz, 1H), 8.60 (s, 1H), 8.42 (s, 2H, D₂O exchangeable), 8.25 (d, J = 6.7 Hz, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.87 (d, J = 17.3 Hz, 1H), 5.48 (d, J = 17.2 Hz, 1H), 4.39 (dd, J 5 9.0, 5.8 Hz, 1H), 3.68 (dd, J = 5.5, 2.4 Hz, 1H), 2.49 - 2.42 (m, 1H), 2.05 - 1.92 (m, 1H), 1.31 (s, 3H), 1.04 (t, J = 5.5 Hz, 1H), 1.01 - 0.94 (m, 1H); MS (ES+): 521/523 (M+l), (ES-): 519/521 (M-l).

Scheme 182

Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pynmidm-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (182a)

Compound 182a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-9H-pyrido[4^,,3^,:4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (29e) (100 mg, 0.280 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N (6 bromopyridin-2-_yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (102 mg, 0.308 mmol), HATU (160 mg, 0.420 mmol), DIPEA (181 mg, 1.400 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 to 7%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-9H-p_yrido[4',3':4,5]p_yrrolo[2,3-d]pyrimidin9-_yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (182a) (87 mg, 60% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.80 (s, 1H, D₂O exchangeable), 9.48 (s, 1H), 9.07 (d, J = 6.3 Hz, 1H), 8.93 - 8.53 (m, 4H, 2H

D₂O exchangeable), 8.01 (d, J = 8.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.89 (d, J = 17.4 Hz, 1H), 5.47 (d, J = 17.2 Hz, 1H), 4.40 (dd, J = 9.1, 5.9 Hz, 1H), 3.65

336 (dd, J = 5.4, 2.6 Hz, 1H), 2.48 - 2.42 (m, 1H), 1.98 (dd, J = 13.3, 5.7 Hz, 1H), 1.31 (s, 3H), 1.08 - 0.97 (m, 2H); MS (ES+): 521/523 (M+l), (ES-): 519/521 (M-l).

Scheme 183

162f

183a

Préparation of (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[3',2^,:4,5]pyrrolo[2,3-d]pynmidm-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (183a)

Compound 183a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (162f) (100 mg, 0.280 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (102 mg, 0.308 mmol), HATU (160 mg, 0.420 mmol), DIPEA (181 mg, 1.400 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 to 7%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-9H-pyrido[3',2^,:4,5]pyrrolo[2,3-d]pynmidin9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (183a) (110 mg, 75% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.77 (s, 1H, D₂O exchangeable), 8.97 (dd, J = 7.9,1.5 Hz, 1H), 8.74 (s, 2H, D2O exchangeable), 8.66 (s, 1H), 8.56 (dd, J = 4.9,1.5 Hz, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.49 (dd, J = 7.9, 4.9 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.71 (d, J = 17.0 Hz, 1H), 5.34 (d, J = 16.9 Hz, 1H), 4.37 (dd, J = 9.0, 5.9 Hz, 1H), 3.72 (dd, J = 5.4, 2.3 Hz, 1H), 2.48 - 2.42 (m, 1H), 1.98 (dd, J = 13.2, 5.9 Hz, 1H), 1.31 (s, 3H), 1.05 (t, J = 5.5 Hz, 1H), 0 86 (dd, J = 5.4, 2.3 Hz, 1H); MS (ES+): 521/523 (M+l), (ES-): 519/521 (M-l); Analysis calculated for C23H2rBrN3O2.LlHCLl.5H2O: C, 46.94; H, 4.30; Cl, 6.63; N, 19.04; Found: C, 46.81; H, 4.17; Cl, 6.66; N, 18.90.

337

Scheme 184

184f 184g

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-35 carboxamide (184g)

Step-1: Préparation of N-(2-bromo-6-methyl-4-(trifluoromethÿl)phenyl)-2,2,2trifluoroacetamide (184b)

Compound 184b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-6-methyl-4-(trifluoromethyl)aniline (184a) (5.34 g, 21.02 mmol; CAS #

1100212-65-4) in DCM (30 mL) using triethylamine (3.62 g, 35.7 mmol), trifluoroacetic acid anhydride (6.62 g, 31.5 mmol) and stirring at RT for 1 h. This gave after workup N-(2bromo-6-methyl-4-(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (184b) (7.50 g, 102% yield) as a pale-orange solid.; ¹H NMR (300 MHz, DMSO-dô) δ 11.55 (s, 1H), 8.03 (d, J = 2.0 Hz, 1H), 7.83 (d, J = 2.0 Hz, 1H), 2.31 (s, 3H). ¹⁹F NMR (282 MHz, DMSO-ώ) δ -61.20, 75 -74.06; MS (ES-): 348/350 (M-l).

Step-2: Préparation of 2-amino-7-methyl-5-(trifluoromethyl)-lH-indole-3-carbonitrile (184c)

338

Compound 184c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-6-methyl-4-(trifluoromethyl)phenyI)-2,2,2-trifluoroacetamide (184b) (7.50 g, 21.43 mmol) in DMSO (25 mL) using malononitrile (1.698 g, 25.7 mmol), L-proline (0.493 g, 4.29 mmol), Cul (0.408 g, 0.2.143 mmol), K2CO3 (5.92 g, 42.9 mmol) in water (25 5 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [SiCh gel (80 g), eluting with EtOAc in hexane from 0-40%] to provide 2-amino7-methyl-5-(trifhioromethyl)-lH-indole-3-carbonitrile (184c) (3.68 g, 72% yield) as a palegreen solid; Ή NMR (300 MHz, DMSO-ifc) δ 11.23 (s, 1H), 7.30 - 7.17 (m, 1H), 7.07 (d, J = 1.8 Hz, 1H), 6.84 (s, 2H), 2.41 (s, 3H). ¹⁹F NMR (282 MHz, DMSO-tfc) δ -58.92; MS (ES+) 10 240 (M+l); (ES-): 238 (M-l).

Step-3: Préparation of 8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-4-amine (184d)

Compound 184d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-7-methyl-5-(trifluoromethyl)-lH-indole-3-carbonitrile (184c) (3.68 g, 15.38 mmol) in éthanol (10 mL) using formamidine acetate (8.01 g, 77 mmol) and NH4OAC (3.56 15 g, 46.2 mmol) and heating at 90 °C for 16 h. This gave after work up 8-methyl-6(trifhioromethyl)-9H-pyrimido[4,5-b]indol-4-amine (184d) (2.69 g, 66% yield) as a tan solid; Ή NMR (300 MHz, DMSO-î/₆) δ 12.26 (s, 1H), 8.60 (s, 1H), 8.32 (s, 1H), 7.48 (d, J = 1.6 Hz, 1H), 7.41 (s, 2H), 2.59 (s, 3H); ¹⁹F NMR (282 MHz, DMSCM₆) δ -58.08; MS (ES+): 267 (M+l), (ES-): 265 (M-l).

Step-4: Préparation of tert-butyl 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetate (184e)

Compound 184e was prepared according to the procedure reported in step-1 of scheme-1, from 8-methyl-6-(trifhioromethyl)-9H-pyrimido[4,5-b]indol-4-amine (184d) (0.56 g, 2.104 mmol) in DMF (10 mL) using ieri-butyl 2-bromoacetate (0.492 g, 2.52 mmol) CS2CO3 (1.371 25 g, 4.21 mmol) and stirring at RT for 16 h. This gave after work up and purification [SiChgel (24 g), eluting with methanol in DCM from 0-6%] /eri-butyl 2-(4-amino-8-methyl-6(trifhioromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (184e) (0.74 g, 92% yield) as a paleyellow solid; MS (ES+): 381 (M+l), (ES-): 379 (M-l).

Step-5: Préparation of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-930 yl)acetic acid (184f)

Compound 184f was prepared according to the procedure reported in step-2 of scheme-1, from teri-butyl 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-921436

339

yl)acetate (184e) (0.74 g, 1.946 mmol) using TFA (2.218 g, 19.46 mmol) in DCM (15 mL) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-8-methyl-6(trifhioromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (0.88 g, 103 % yield) as a pale-yellow solid; MS (ES+): 325 (M+l), (ES-): 323 (M-l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (184g)

Compound 184g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-910 yl)acetic acid (184f) (100 mg, 0.228 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (83 mg, 0.251 mmol), HATU (130 mg, 0.342 mmol) DIPEA (147 mg, 1.14 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 to 2%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (184g) (76 mg, 55% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.83 (s, 1H, D₂O exchangeable), 8.79 (s, 1H), 8.72 (s, 2H, 20 D₂O exchangeable), 8.64 (s, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.60 (s, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.92 (d, J = 18.0 Hz, 1H), 5.64 (d, J = 17.9 Hz, 1H), 4.38 (dd, J = 9.0, 6.1 Hz, 1H), 3.71 (dd, J = 5.6, 2.4 Hz, 1H), 2.75 (s, 3H), 2.48 - 2.44 (m, 1H), 1.97 (dd, J = 13.2, 6.0 Hz, 1H), 1.30 (s, 3H), 1.02 (t, J = 5.4 Hz, 1H), 0.85 (dd, J = 5.4, 2.4 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -58.78; MS (ES+): 602/604 (M+l), (ES-): 600/602 (M-l).

²⁵ Scheme 185

340

Préparation of (lR,3S,5R)-2-(2-(6-acetamido-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (185a)

Compound 185a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(6-acetamido-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (1681) (75 mg, 0.181 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (66.4 mg, 0.200 mmol), HATU (103 mg, 0.272 mmol) DIPEA (117 mg, 0.907 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 to 7%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(6-acetamido-4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (185a) (57 mg, 54% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ 10.78 (s, 1H, D₂O exchangeable), 10.06 (s, 1H, D₂O exchangeable), 8.82 - 8.52 (m, 3H, 2H D₂O exchangeable), 8.49 (d, J = 1.9 Hz, 1H), 8.01 (d, J = 8.1 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.62 (d, J = 8.8 Hz, 1H), 7.53 (dd, J = 8.8,1.8 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.70 (d, J = 17.3 Hz, 1H), 5.34 (d, J = 17.3 Hz, 1H), 4.37 (dd, J = 9.1, 5.9 Hz, 1H), 3.68 (dd, J = 5.5, 2.4 Hz, 1H), 2.47 - 2.41 (m, 1H), 2.08 (s, 3H), 1.97 (dd, J = 13.3, 5.9 Hz, 1H), 1.30 (s, 3H), 1.00 (t, J = 5.5 Hz, 1H), 0.93 (dd, J = 5.4, 2.4 Hz, 1H); MS (ES+): 577/579 (M+l); (ES-): 575/577 (M-l). .

Scheme 186

35e

186a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (186a)

Compound 186a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (35e) (75

341 .

mg, 0.172 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (57.3 mg, 0.172 mmol), HATU (79 mg, 0.207 mmol), DIPEA (111 mg, 0.862 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with

MeOH in DCM from 0 to 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (186a) (69 mg, 67% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.77 (s, 1H, D₂O exchangeable), 8.57 (s, 1H), 8.49 (d, J = 7.9 Hz, 1H), 8.37 (s, 2H, D₂O exchangeable), 8.01 (d, J = 8.2 Hz, 1H), 7.75 - 7.61 (m, 2H), 7.29 (t, J = 8.0 Hz, 2H), 5.93 (d, J = 17.7 Hz, 1H), 5.72 (d, J = 1Ί.6 Hz, 1H), 4.36 (dd, J = 8.9, 6.1 Hz, 1H), 3.66 (dd, J = 5.5, 2.4 Hz, 1H), 2.43 (d, J = 6.6 Hz, 1H), 1.97 (dd, J = 13.1, 5.9 Hz, 1H), 1.30 (s, 3H), 1.00 (t, J = 5.5 Hz, 1H), 0.89 (dd, J = 5.6, 2.3 Hz, 1H); MS (ES+): 598/600 (M+l), 596/598 (M-l).

¹⁵ Scheme 187

Préparation of(lR,3S,5R)-2-(2-(4-amino-6-ethyl-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(6. bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (187c)

Step-1: Préparation of ethyl 2-(4-amino-6-ethyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (187a)

A solution of ethyl 2-(4-amino-6-vinyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (59a) (100 mg, 0.337 mmol) in ethyl acetate (5 mL) and éthanol (5.00 mL) was treated with palladium (53.9 mg, 0.051 mmol) and hydrogenated for 2 h. The reaction mixture was filtered, washed with ethyl acetate/ethanol. The filtrate was concentrated to afford ethyl 2-(4-amino-6-ethyl-9Hpyrimido[4,5-b]indol-9-yl)acetate (187a) which was used as such for next step; MS (ES+)·

299.10 (M+l).

Step-2: Préparation of 2-(4.amino.6-ethyl-9H-pyrimido[4,5.b]indol-9-yl)acetic acid (187b)

342

Compound 187b was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-6-ethyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (187a) (67 mg, 0.225 mmol) in THF (3 mL) and MeOH (3 mL) using a solution of lithium hydroxide hydrate (57.7 mg, 1.347 mmol) in water (3 mL) and stirring at RT for 20 h. This gave after workup 2-(45 amino-6-ethyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (187b) which used as such for the next step; MS (ES+): 271.10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-ethyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (187c)

Compound 187c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-ethyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (187b) (61 mg, 0.225 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (108 mg, 0.338 mmol), HATU (171 mg, 0.450 mmol), DIPEA (0.157 mL, 0.900 mmol) and stirring at RT for 18 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with dichloromethane/methanol (1:0 to 19:1)] followed by purification using reverse phase column chromatography [C18 column, eluting with ACN in water (containing 0.1% HCl) from 0100%] (lR,3S,5R)-2-(2-(4-amino-6-ethyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (187c) (25 mg, 21% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-rf₆) δ 10.75 (s, 1H), 8.58 (s, 1H), 8.52 (s, 2H), 8.35 (s, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.60 (d, J = 8.4 Hz,

1H), 7.45 - 7.39 (m, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.73 (d, J = 17.3 Hz, 1H), 5.40 (d, J = 17.3 Hz, 1H), 4.40 (dd, J = 9.0, 5.5 Hz, 1H), 4.04 - 3.78 (m, 1H), 2.78 (q, J = 7.6 Hz, 2H), 2.41 - 2.13 (m, 2H), 2.00 - 1.78 (m, 1H), 1.28 (t, J = 7.5 Hz, 3H), 1.12 - 0.98 (m, 1H), 0.82 0.67 (m, 1H); MS (ES+): 534.10 & 536.10 (M+l); MS (ES-): 532.00 & 534.10 (M-l);

Analysis calculated for C25H₂4BrN₇O₂.HC1.2.25H2O: C, 49.11; H, 4.86; N, 16.04; Found: C, 49.18; H, 4.52; N, 15.67. ’

Scheme 188

343

188d 188e

188f

188g ' Préparation of (lR,3S,5R)-2-(2-(4-amino-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (188g)

Step-1: Préparation of 2,2,2-trifluoro-N-(2-iodo-4-nitrophenyl)acetamide (188b)

Compound 188b was prepared according to the procedure reported in step-1 of scheme-46, from 2-iodo-4-nitroaniline (188a) (5 g, 18.94 mmol; CAS # 6293-83-0) in DCM (35 mL) using triethylamine (6.60 mL, 47.3 mmol), trifluoroacetic acid anhydride (3.95 mL, 28.4 mmol) and stirring at RT for 19 h. This gave after workup 2,2,2-trifluoro-N-(2-iodo-4nitrophenyl)acetamide (188b) (8.986 g) as a brown gum which was used as such for the next 10 step; (ES-): 358.90 (M-l).

Step-2: Préparation of 2-amino-5-nitro-lH-indole-3-carbonitrile (188c)

Compound 188c was prepared according to the procedure reported in step-1 of scheme-11, from 2,2,2-trifluoro-N-(2-iodo-4-nitrophenyl)acetamide (188b) (6.819 g, 18.94 mmol) in DMSO (25 mL) using malononitrile (1.501 g, 22.73 mmol), L-proline (0.436 g, 3.79 mmol),

344

Cul (0.361 g, 1.894 mmol), a solution of K2CO3 (5.235 g, 37.9 mmol) in water (25 mL) and heatmg at 60 °C for 13 h under an argon atmosphère. This gave after workup and purification [SiChgel (120 g), eluting with EtOAc in hexane from 0-67%] 2-amino-5-nitro-lH-indole-3carbonitrile (188c) (531 mg, 14% yield) as a red solid; ^XH NMR (300 MHz, DMSO-de) δ 11.43 (s, 1H), 7.92 (d, J = 2.3 Hz, 1H), 7.85 (dd, J = 8.7; 2.3 Hz, 1H), 7.34 (s, 2H), 7.28 (d, J = 8.7 Hz, 1H); (ES+): 203.05 (M+l); (ES-): 201.00 (M-l).

Step-3: Préparation of 6-nitro-9H-pyrimido[4,5-b]indol-4-amine (188d)

Compound 188d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5-nitro-lH-indole-3-carbonitrile (188c) (520 mg, 2.57 mmol) in éthanol (12 mL) using formamidine acetate (2.164 g, 20.58 mmol) and refluxing for 44 h. This gave after work up 6-nitro-9H-pyrimido[4,5-b]indol-4-amine (188d) (497 mg, 84% yield) as a gray solid; Ή NMR (300 MHz, DMSOX) δ 12.54 (s, 1H), 9.34 (d, J = 2.2 Hz, 1H), 8.33 (s, 1H), 8.26 (dd, J = 8.9, 2.2 Hz, 1H), 7.62 (s, 2H), 7.58 (d, J = 9.0 Hz, 1H); (ES+): 230.10 (M+l); (ES-): 228.05 (M-l).

Step-4: Préparation of ethyl 2-(4-amino-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetate (188e)

Compound 188e was prepared according to the procedure reported in step-1 of scheme-1, from 6-nitro-9H-pyrimido[4,5-b]indol-4-amine (188d) (300 mg, 1.309 mmol) in DMF (15 mL) using ethyl 2-bromoacetate (0.160 mL, 1.440 mmol), CS2CO3 (1.066 g, 3.27 mmol) and stirring at RT for 18 h. This gave after work up ethyl 2-(4-amino-6-nitro-9H-pyrimido[4,5b]indol-9-yl)acetate (188e) (0.373 g, 90% yield) as a yellow solid; ^XH NMR (300 MHz, DMSO-Jô) δ 9.39 (d, J = 2.2 Hz, 1H), 8.38 (s, 1H), 8.32 (dd, J = 9.0, 2.2 Hz, 1H), 7.85 (d, J = 9.1 Hz, 1H), 7.78 (s, 2H), 5.35 (s, 2H), 4.15 (q, J = 7.1 Hz, 2H), 1.20 (t, J = 7.1 Hz, 3H); (ES+): 316.15 (M+l); (ES-): 314.00 (M-l).

Step-5: Préparation of 2-(4-amino-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (188f)

Compound 188f was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetate (188e) (0.100 g, 0.317 mmol) in THF (3 mL) and MeOH (3 mL) using a solution of lithium hydroxide hydrate (81 mg, 1.903 mmol) in water (3 mL) and stirring at RT for 22 h. This gave after work up 2-(4amino-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (188f) which was used as such for the next step; (ES+): 287.80 (M+l).

345

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-nitro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (188g)

Compound 188g was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (188f) (61 mg, 0.225 mmol) in DMF (12 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (151 mg, 0.476 mmol), HATU (241 mg, 0.634 mmol), DIPEA (0.276 mL, 1.585 mmol) and stirring at RT for 15 h. This gave after workup and purification by reverse phase column chromatography [C18 column, eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-nitro-9H10 pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (188g) (50 mg, 29% yield) HCl sait as a white solid; ³H NMR (300 MHz, DMSO-de) δ 10.76 (s, 1H), 9.52 (d, J = 2.2 Hz, 1H), 8.73 (s, 3H), 8.64 (s, 1H), 8.41 (dd, J = 9.1, 2.2 Hz, 1H), 8.00 (d, J = 8.1 Hz, 1H), 7.88 (d, J = 9.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.84 (d, J = 17.4 Hz, 1H), 5.48 (d, J = 17.3 Hz, 1H), 4.42 (dd, J =

9.0, 5.5 Hz, 1H), 3.97 - 3.86 (m, 1H), 2.44 - 2.12 (m, 2H), 1.99 - 1.79 (m, 1H), 1.17 - 0.96 (m, 1H), 0.88 - 0.67 (m, 1H); (ES+): 551.10 & 553.10 (M+l); (ES-): 549.00 & 551.00 (M-l); Analysis calculated for C23Hi9BrN₈O4.0.85HC1.1.75H₂O: C, 45.00; H, 3.83; N, 18.25; Cl, 4.91; Found: C, 45.22; H, 3.59; N, 17.88; Cl, 4.90.

Scheme 189

346

189d

189e

Préparation of (lR,3S,5R)-2-(2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3 1.0]hexane-3-carboxamide (189g)

Step-1: Préparation of N-(2-bromo-4,6-dimethylphenyl)-2,2,2-trifluoroacetamide (189b)

Compound 189b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-4,6-dimethylaniline (189a) (7 g, 35.0 mmol; CAS # 41825-73-4) in DCM (30 mL) using tnethylamine (6.02 g, 59.5 mmol), trifluoroacetic acid anhydride (11.02 g, 52.5 mmol) and stirring at RT for 1 h. This gave after workup N-(2-bromo-4,6-dimethylphenyl)2,2,2-trifhioroacetamide (189b) (10.32 g, 100% yield) as a pale-yellow solid; Ή NMR (300

MHz, DMSO-dô) δ 11.12 (s, 1H), 7.43 (s, 1H), 7.18 (s, 1H), 2.30 (s, 3H), 2.16 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-flfe) δ -74.08; MS (ES+): 296/298 (M+l); (ES-): 294/296 (M-l). Step-2: Préparation of 2-amino-5,7-dimethyl-lH-indole-3-carbonitrile (189c)

Compound 189c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-4,6-dimethylphenyl)-2,2,2-trifluoroacetamide (189b) (10.32 g, 34.9 mmol) in DMSO (30 mL), using malononitrile (2.76 g, 41.8 mmol), L-proline (0.803 g, 6.97 mmol), Cul (0.664 g, 3.49 mmol), and a solution of K₂CO₃ (9.63 g, 69.7 mmol) in water (30 mL) and , 347 ' heatmg at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [SiO₂gel (40 g), eluting with EtOAc in hexane from 0-40%] 2-amino-5,7-dimethyl-lHindole-3-carbonitrile (189c) (3.97 g, 62% yield) as a pale-orange solid; 'H NMR (300 MHz, DMSO-î/ô) δ 10.60 (s, 1H), 6.77 (s, 1H), 6.54 (d, J = 1.6 Hz, 1H), 6.43 (s, 2H), 2.28 (s, 3H), 5 2.27 (s, 3H); MS (ES+): 186 (M+l); (ES-): 184 (Μ-1).

Step-3: Préparation of 6,8-dimethyl-9H-pyrimido[4,5-b]indol-4-amine (189d)

Compound 189d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5,7-dimethyl-lH-indole-3-carbonitrile (189c) (3.97 g, 21.43 mmol) in éthanol (10 mL) using trimethoxymethane (11.37 g, 107 mmol), NH₄OAc (4.96 g, 64.3 mmol) and heatmg at 90 °C for 16 h. This gave after work up 6,8-dimethyl-9H-pyrimido[4,5-b]indol-4amine (189d) (4.07 g, 89% yield) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-î/₆) δ 11.68 (s, 1H), 8.22 (s, 1H), 7.93 (s, 1H), 7.03 (s, 2H), 6.98 (s, 1H), 2.47 (s, 3H), 2.42 (s, 3H); MS (ES+): 213 (M+l), (ES-): 211 (M-l). ’ ’

Step-4: Préparation of terAbutyl 2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-915 yl)acetate (189e)

Compound 189e was prepared according to the procedure reported in step-1 of scheme-1, from 6,8-dimethyl-9H-pyrimido[4,5-b]indol-4-amine (189d) (2.00 g, 9.42 mmol) in DMF (15 mL) using tert-butyl 2-bromoacetate (2.206 g, 11.31 mmol), Cs₂CO₃ (6.14 g, 18.85 mmol) and stirring at RT for 16 h. This gave after work up terAbutyl 2-(4-amino-6,8-dimethyl-9H20 pyrimido[4,5-b]indol-9-yl)acetate (189e) (1.79 g, 58%) as a tan solid which was used as such for the next step; Ή NMR (300 MHz, DMSO-J₆) δ 8.26 (s, 1H), 8.02 (s, 1H), 7.19 (s, 2H), 6.98 (s, 1H), 5.29 (s, 2H), 2.59 (s, 3H), 2.41 (s, 3H), 1.42 (s, 9H); MS (ES+): 327 (M+l). Step-5: Préparation of 2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (189f)

Compound 189f was prepared according to the procedure reported in step-2 of scheme-1, from Zeri-butyl 2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (189e) (300 mg, 0.919 mmol) using TFA (1048 mg, 9.19 mmol) in DCM (15 mL) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol9-yl)acetic acid (189f) (0.366 g, 104% yield) as a pale-yellow solid; ’H NMR (300 MHz,

DMSO-^) δ 8.66 (s, 2H), 8.61 (s, 1H), 8.19 (s, 1H), 7.16 (s, 1H), 5.43 (s, 2H), 2.65 (s, 3H), 2.45 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-rf₆) δ -74.52; MS (ES+): 271 (M+l)· (ES-)· 269 ’ (M-l). ’

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (189g)

Compound 189g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (1891) (75 mg, 0.195 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (62.2 mg, 0.195 mmol), HATU (89 mg, 0.234 mmol), DIPEA (126 mg, 0.976 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography

[C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (189g) (75 mg, 72% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.82 (s, 1H, D₂O exchangeable), 8.68 (s, 2H, D₂O exchangeable), 8.62 (s, 1H), 8.17 (s, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 7.11 (s, 1H), 5.88 (d, J = 17.9 Hz, 1H), 5.60 (d, J =

17.8 Hz, 1H), 4.41 (dd, J = 9.1, 5.7 Hz, 1H), 3.91 (ddd, J = 7.3, 5.4, 2.4 Hz, 1H), 2.66 (s, 3H), 2.42 (s, 3H), 2.40 - 2.29 (m, 1H), 2.26 - 2.14 (m, 1H), 1.99 - 1.85 (m, 1H), 1.14 - 0.98 (m, 1H), 0.74 - 0.62 (m, 1H); MS (ES+): 534/536 (M+l), (ES-): 532/534 (M-l); Analysis calculated for C₂₅H₂4BrN₇O₂.HC1.1.75H₂O: C, 49.85; H, 4.77; Cl, 5.89; N, 16.28; Found: C,

49.82; H, 4.65; Cl, 5.83; N, 16.03. ’ ’

Scheme 190

190a

Préparation of ethyl 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-broniopyri_<iin-2-yl)carba_moyl)-5methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-625 yl)propanoate (190a)

349

Compound 190a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(3-ethoxy-3-oxopropyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (45c) (75 mg, 0.164 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (65.6 mg, 0.197 5 mmol), HATU (94 mg, 0.247 mmol) DIPEA (106 mg, 0.822 mmol) and stirring at RT for 16

h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] ethyl 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)10 5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6yl)propanoate (190a) (78 mg, 76% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-de) δ 10.77 (s, 1H, D₂O exchangeable), 8.80 - 8.54 (m, 3H, 2H D₂O exchangeable), 8.46 - 8.36 (m, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.61 (d, J = 8.5 Hz, ’ 1H), 7.45 (dd, J = 8.5, 1.5 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.70 (d, J = 17.3 Hz, 1H), 5.35 (d, J = 17.2 Hz, 1H), 4.37 (dd, J = 9.0, 6.0 Hz, 1H), 4.05 (q, J = 7.1 Hz, 2H), 3.69 (dd, J = 5 6

2.4 Hz, 1H), 3.03 (t, J = 7.7 Hz, 2H), 2.75 (t, J = 7.7 Hz, 2H), 2.49 - 2.40 (m, 1H), 2.05 -1.92 (m, 1H), 1.31 (s, 3H), 1.16 (t, J = 7.1 Hz, 3H), 1.01 (t, J = 5.5 Hz, 1H), 0.97 - 0.88 (m, 1H);

MS (ES+): 620/622 (M+l), (ES-): 618/620 (M-l); Analysis calculated for ’

C₂₉H₃oBrN₇0₄.HC1.1.5H₂0: C, 50.92; H, 5.01; N, 14.33; Found: C, 50.82; H, 4.90; N, 14.05.

²⁰ Scheme 191

Préparation of(lR,3S,5R)-2-(2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5.methyl.2-azabicyclo[3.1.0]hexane-3-carboxamide (191a)

Compound 191a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6,8-dimethyl.9H.pyrimido[4,5-b]mdol-9-yl)acetic acid (1891)

350 (75 mg, 0.195 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (64.9 mg, 0.195 mmol), HATU (89 mg, 0.234 mmol), DIPEA (126 mg, 0.976 mmol)and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with

MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (191a) (63 mg, 59% yield) HCl as a white solid; Ή NMR (300 MHz, DMSO-t/₆) δ 10.80 (s,

1H, D2O exchangeable), 8.70 (s, 2H, D2O exchangeable), 8.61 (s, 1H), 8.16 (s, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.30 (d, J = 7.7 Hz, 1H), 7.11 (s, 1H), 5.82 (d, J = 18.0 Hz, 1H), 5.55 (d, J = 17.9 Hz, 1H), 4.37 (dd, J = 9.0, 6.2 Hz, 1H), 3.69 - 3.67 (m, 1H), 2.63 (s, 3H), 2.48 - 2.33 (m, 4H), 1.97 (dd, J = 13.2, 6.1 Hz, 1H), 1.30 (s, 3H), 1.01 (t, J = 5.5 Hz, 1H), 0.83 (dd, J = 5.4, 2.3 Hz, 1H); MS (ES+) 548/550 (M+l), 546/548 (M-l); Analysis calculated for C26H26BrN7O2.HCl.2H2O: C, 50.29; H, 5.03; Cl, 5.71; N, 15.79; Found: C, 50.14; H, 4.95; Cl, 5.64; N, 15.56.

Scheme 192

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-920 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (192g)

Step-1: Préparation of N-(2,4-dibromo-6-methylphenyl)-2,2,2-trifluoroacetamide (192b)

Compound 192b was prepared according to the procedure reported in step-1 of scheme-46, ' from 2,4-dibromo-6-methylaniline (192a) (10 g, 37.7 mmol; CAS # 30273-41-7) in DCM (30'

351 mL) using triethylamine (6.49 g, 64.2 mmol), trifluoroacetic acid anhydride (11.89 g, 56.6 mmol) and stirring at RT for 1 h. This gave after workup N-(2,4-dibromo-6-methylphenyl)2,2,2-trifluoroacetamide (192b) (13.29 g, 98% yield) as a purple solid; *H NMR (300 MHz, DMSO-cZô) δ 11.30 (s, 1H), 7.88 (s, 1H), 7.66 (s, 1H), 2.21 (s, 3H); ¹⁹F NMR (282 MHz, 5 DMSO-de) δ -74.09; MS (ES-): 358/360 (M-l).

Step-2: Préparation of 2-amino-5-bromo-7-methyl-lH-indole-3-carbonitrile (192c)

Compound 192c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2,4-dibromo-6-methylphenyl)-2,2,2-trifluoroacetamide (192b) (13.29 g, 36.8 mmol) in DMSO (20 mL) using malononitrile (2.92 g, 44.2 mmol), L-proline (0.848 g, 7.36 mmol), 10 Cul (0.701 g, 3.68 mmol), a solution of K2CO3 (10.18 g, 73.6 mmol) in water (30 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [SiO₂gel (40 g), eluting with MeOH in DCM from 0-6%] 2-amino-5-bromo-7-methyl-lHindole-3-carbonitrile (192c) (5.54 g, 60% yield) as a deep green-black solid; ^]H NMR (300 MHz, DMSO-ifc) δ 10.97 (s, 1H), 7.06 (d, J = 1.9 Hz, 1H), 6.89 (d, J = 1.9 Hz, 1H), 6.71 (s, 15 2H), 2.32 (s, 3H); MS (ES+): 250/252 (M+l); (ES-): 248/250 (M-l).

Step-3: Préparation of 6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-4-amine (192d)

Compound 192d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5-bromo-7-methyl-lH-indole-3-carbonitrile (192c) (5.54 g, 22.15 mmol) in éthanol (10 mL) using trimethoxymethane (11.75 g, 111 mmol), NH4OAC (5.12 g, 66.5 20 mmol) and heating at 80 °C for 16 h. This gave after work up 6-bromo-8-methyl-9Hpyrimido[4,5-b]indol-4-amine (192d) (5.57 g, 91% yield) as a green solid; *H NMR (300 MHz, DMSO-rf₆) δ 12.01 (s, 1H), 8.40 (d, J = 1.8 Hz, 1H), 8.27 (s, 1H), 7.33 (dd, J = 1.9, 0.9 Hz, 1H), 7.26 (s, 2H), 2.50 (d, J = 2.2 Hz, 3H); MS (ES+): 277/279 (M+l), (ES-): 275/277 (M-l).

Step-4: Préparation of ZerZ-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetate (192e)

Compound 192e was prepared according to the procedure reported in step-1 of scheme-1, from 6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-4-amine (192d) (2.00 g, 7.22 mmol) in DMF (15 mL) using ZerZ-butyl 2-bromoacetate (1.689 g, 8.66 mmol), Cs₂CO3 (4.70 g, 14.43 30 mmol) and stirring at RT for 16 h. This gave after work up and purification [SiO₂gel (40 g), eluting with methanol in DCM from 0-4%] ZerZ-butyl 2-(4-amino-6-bromo-8-methyl-9Hpyrimido[4,5-b]indol-9-yl)acetate (192e) (1.65 g, 58% yield) as a tan solid; Ή NMR (300

352

MHz, DMSO-d₆) δ 8.47 (d, J = 1.9 Hz, 1H), 8.31 (s, 1H), 7.43 (s, 2H), 7.38 - 7.30 (m, 1H), 5.33 (s, 2H), 2.62 (s, 3H), 1.42 (s, 9H); MS (ES+): 391/393 (M+l), (ES-): 389/391 (M-l). Step-5: Préparation of 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (192f)

Compound 192f was prepared according to the procedure reported in step-2 of scheme-1, from iert-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (192e) (300 mg, 0.767 mmol) using TFA (874 mg, 7.67 mmol) in DCM (15 mL) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-6-bromo-8-methyl-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (192f) (0.357 g) as a pale-yellow solid; ’H NMR (300 10 MHz, DMSO-tfc) δ 8.62 (d, J = 1.9 Hz, 1H), 8.55 (s, 1H), 8.45 (s, 2H), 7.48 (d, J = 1.9 Hz, 1H), 5.42 (s, 2H), 2.67 (s, 3H); ’⁹F NMR (282 MHz, DMSO-ife) δ -74.55; MS (ES+): 335/337 (M+l); (ES-): 333/335 (M-l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide 15 (192g)

Compound 192g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (1921) (75 mg, 0.167 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (53.2 mg, 0.167 mmol), 20 HATU (76 mg, 0.200 mmol), DIPEA (108 mg, 0.835 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-925 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (192g) (83 mg, 83% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-ifo) δ 10.83 (s, 1H, D2O exchangeable), 8.76 (s, 2H, D₂O exchangeable), 8.68 - 8.56 (m, 2H), 8.00 (d, J = 8.1 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.48 (d, J = 1.9 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.90 (d, J = 18.0 Hz, 1H), 5.63 (d, J = 17.9 Hz, 1H), 4.42 (dd, J = 9.0, 5.7 Hz, 1H), 3.91 (td, J = 5.4, 3.0

Hz, 1H), 2.69 (s, 3H), 2.41 - 2.29 (m, 1H), 2.26 - 2.10 (m, 1H), 2.00 - 1.83 (m, 1H), 1.18 0.97 (m, 1H), 0.75 - 0.60 (m, 1H); MS (ES+): 598/600 (M+l); (ES-): 596/598 (M-l);

Analysis calculated for C24H2iBr₂N7O2.1.1HC1.1.75H₂O: C, 42.96; H, 3.85; Cl, 5.81; N, 14.61; Found: C, 43.08; H, 3.70; Cl, 5.91; N, 14.34.

353

Scheme 193

Br ^192f 193a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide 5 (193a)

Compound 193a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (192f) (75 mg, 0.167 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (55.5 mg, 0.167 mmol), HATU (76 mg, 0.200 mmol), DIPEA (108 mg, 0.835 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol15 9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (193a) (83 mg, 81% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.82 (s, 1H, D₂O exchangeable), 8.60 (d, J = 1.9 Hz, 1H), 8.57 (s, 1H), 8.51 (s, 2H, D₂O exchangeable), 8.01 (d, J = 8.2 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.45 (d, J = 1.8 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.84 (d, J = 18.0 Hz, 1H), 5.58 (d, J = 17.9 Hz, 1H), 4.37 (dd, J =

9.0, 6.1 Hz, 1H), 3.70 - 3.66 (m, 1H), 2.67 (s, 3H), 2.48 - 2.40 (m, 1H), 1.97 (dd, J = 13.2, 6.1

Hz, 1H), 1.30 (s, 3H), 1.01 (t, J = 5.5 Hz, 1H), 0.83 (dd, J = 5.4, 2.4 Hz, 1H); MS (ES+): 612/614 (M+l), (ES-): 610/612 (M-l); Analysis calculated for C₂5H₂₃Br2N₇O₂.HC1.1.75H₂O: C, 44.07; H, 4.07; Cl, 5.20; N, 14.39; Found: C, 44.02; H, 3.91; Cl, 5.46; N, 14.13.

Scheme 194

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (194a)

Compound 194a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (1840 (100 mg, 0.228 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (80 mg, 0.251 mmol), HATU (130 mg, 0.342 mmol), DIPEA (147 mg, 1.141 mmol)and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 2%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (194a) (33 mg, 25% yield) HCl sait as a white solid; ^XH NMR (300 MHz, DMSO-î/ô) δ 10.83 (s, 1H, D₂O exchangeable), 8.78 (s, 1H), 8.59 (s, 1H), 8.50 (s, 2H, D₂O exchangeable), 8.01 (d, J = 8.2 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.64 - 7.57 (m, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.96 (d, J = 18.0 Hz, 1H), 5.69 (d, J = 17.9 Hz, 1H), 4.43 (dd, J = 9.1, 5.6 Hz, 1H), 3.97 - 3.92 (m, 1H), 2.78 (s, 3H), 2.40 - 2.30 (m, 1H), 2.30 - 2.15 (m, 1H), 2.00 - 1.86 (m, 1H), 1.15 - 1.04 (m, 1H), 0.74

- 0.64 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-tfc) δ -58.71; MS (ES+): 588/590 (M+l), (ES-):

586/588 (M-l).

Scheme 195

355

HATL), DIPEA

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(hydroxymeth_yl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (195e)

Step-1: Préparation of ethyl 2-(4-amino-6-(l,2-dih_ydroxyethyl)-9H-pyrimido[4,5-b]indol-95 yl)acetate (195a) .

To a solution of ethyl 2-(4-amino-6-vinyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (59a) (300 mg, 1.012 mmol) in acetonitrile (20 mL) and water (10 mL) was added 4-methylmorpholine 4-oxide (232 mg, 1.924 mmol) followed by osmium(VIII) oxide (4 % in water) (0.619 mL, 0.101 mmol) and stirred at RT for 22 h. Reaction mixture was quenched with 1 M aqueous

Na2S2Û3 solution (10 mL), diluted with water (50 mL), and extracted with ethyl acetate (100 mL, 2 x 75 mL). The combined organics were dried, filtered and concentrated in vacuum.

The residue obtained was purified by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM (from 0-10%)] to afford ethyl 2-(4-amino-6-(l,2-dihydroxyethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetate (195a) (122 mg, 37%) as a yellow solid; ’H NMR (300 15 MHz, DMSO-de) δ 8.29 - 8.24 (m, 2H), 7.51 (d, J = 8.4 Hz, 1H), 7.44 - 7.38 (m, 1H), 7.25 (s, 2H), 5.27 - 5.15 (m, 3H), 4.75 - 4.64 (m, 2H), 4.13 (q, J = 7.1 Hz, 2H), 3.55 (t, J = 5.9

Hz, 2H), 1.20 (t, J = 7.1 Hz, 3H).; MS (ES+): 331.15 (M+l); (ES-): 329.10 (M-l).

Step-2: Préparation of ethyl 2-(4-amino-6-formyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (195b)

To a solution of ethyl 2-(4-amino-6-(l,2-dihydroxyethyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (195a) (117 mg, 0.354 mmol) in diethyl ether (10 mL) and water (5 mL) was added

356 sodium periodate (152 mg, 0.708 mmol) and stirred at RT for 1 h. The reaction mixture was diluted with ethyl acetate (100 mL), water (50 mL), 1 M aqueous Na2S2Û3 solution (1 mL) and filtered. The filtrate was separated washed with brine (50 mL), dried, filtered and concentrated in vacuum to afford ethyl 2-(4-amino-6-formyl-9H-pyrimido[4,5-b]indol-95 yl)acetate (195b) as an off-white solid (101 mg) which was used as such for next step; 'H NMR (300 MHz, DMSO-î/₆) δ 10.06 (s, 1H), 9.00 (d, J = 1.5 Hz, 1H), 8.35 (s, 1H), 7.96 (dd, J = 8.6,1.5 Hz, 1H), 7.81 (d, J = 8.5 Hz, 1H), 7.58 (s, 2H), 5.32 (s, 2H), 4.15 (q, J = 7.1 Hz, 2H), 1.20 (t, J = 7.1 Hz, 3H); MS (ES+): 299.10 (M+l).

Step-3: Préparation of methyl/ethyl 2-(4-amino-6-(hydroxymethyl)-9H-pyrimido[4,5-b]indol10 9-yl)acetate (195c)

Compound 195c was prepared according to the procedure reported in scheme-157, from ethyl 2-(4-amino-6-formyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (195b) (0.106 g, 0.354 mmol) in THF (10 mL) and MeOH (5 mL) using sodium borohydride (0.027 g, 0.708 mmol) and stirring at RT for 2 h. This gave after work up a mixture of methyl/ethyl 2-(4-amino-615 (hydroxymethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (195c) as a white solid (90 mg) which was used as such for the next step; MS (ES+): 301.10 & 287.10 (M+l).

Step-4: Préparation of 2-(4-amino-6-(hydroxymethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (195d)

Compound 195d was prepared according to the procedure reported in step-4 of scheme-17, 20 from a mixture of methyl/ethyl 2-(4-amino-6-(hydroxymethyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (195c) (90 mg, 0.3 mmol) in THF (5 mL) and MeOH (5 mL) using a solution of lithium hydroxide hydrate (77 mg, 1.798 mmol) in water (5 mL) and stirring at RT for 20 h.

This gave after work up 2-(4-amino-6-(hydroxymethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (195d) which was used as such for the next step; MS (ES+): 272.80 (M+l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(hydroxymethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (195e)

Compound 195e was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(hydroxymethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (195d) (82 mg, 0.3 mmol) in DMF (12 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (143 mg, 0.450 mmol), HATU (228 mg, 0.600

357 mmol), DIPEA (0.261 mL, 1.500 mmol) and stirring at RT for 15 h. This gave after workup and purification using reverse phase column chromatography [C18 column, eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6(hydroxymethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-25 azabicyclo[3.1.0]hexane-3-carboxamide (195e) (33 mg, 21% yield) HCl sait as a white solid; ‘H NMR (300 MHz, DMSO-ifc) δ 10.76 (s, 1H), 8.58 (s, 1H), 8.52 (s, 3H), 8.44 (s, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.65 (d, J = 8.7 Hz, 1H), 7.58 - 7.51 (m, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.75 (d, J = 17.3 Hz, 1H), 5.41 (d, J = 17.2 Hz, 1H), 4.66 (s, 2H), 4.41 (dd, J = 9.1, 5.6 Hz, 1H), 3.96 - 3.86 (m, 1H), 2.41 - 2.13 (m, 2H), 2.01 - 1.79 (m, 1H), 10 1.14 - 0.99 (m, 1H), 0.87 - 0.59 (m, 1H); MS (ES+): 536.10 & 538.10 (M+l); MS (ES-):

534.10 & 536.10 (M-l); Analysis calculated for C24H22BrN₇O3.1.0HC1.2.75H₂O: C, 46.31; H, 4.62; N, 15.75; Found: C, 46.28; H, 4.29; N, 15.40.

Scheme 196

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-N-methyl-9H-pyrimido[4,5-b]indole-6carboxamide (196d)

Step-1: Préparation of 4-amino-N-methyl-9H-pyrimido[4,5-b]indole-6-carboxamide (196a)

358

Compound 196a was prepared according to the procedure reported in scheme-119, from 4amino-9H-pyrimido[4,5-b]indole-6-carboxylic acid (169a) (200 mg, 0.876 mmol) in DMSO (12 mL) using methanamine hydrochloride (178 mg, 2.63 mmol; CAS # 593-51-1), HOBT (59.2 mg, 0.438 mmol), EDC (336 mg, 1.753 mmol), DIPEA (0.458 mL, 2.63 mmol) and stirring at RT for 20 h. The residue obtained after work up of 4-amino-N-methyl-9Hpyrimido[4,5-b]indole-6-carboxamide (196a) was used as such for the next step; MS (ES+): 241.90 (M+l); (ES-): 239.85 (M-l).

Step-2: Préparation of ZerZ-butyl 2-(4-amino-6-(methylcarbamoyl)-9H-pyrimido[4,5-b]indol9-yl)acetate (196b)

Compound 196b was prepared according to the procedure reported in step-1 of scheme-1, from 4-amino-N-methyl-9H-pyrimido[4,5-b]indole-6-carboxamide (196a) (211 mg, 0.876 mmol) in DMF using ZerZ-butyl 2-bromoacetate, CS2CO3 and stirring at RT. This gave after workup ZerZ-butyl 2-(4-amino-6-(methylcarbamoyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (196b) which was used as such in the next step; MS (ES+): 356.20 (M+l); (ES-): 354.10 (M75 1). ,

Step-3: Préparation of 2-(4-amino-6-(methylcarbamoyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (196c)

Compound 196c was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6-(methylcarbamoyl)-9H-pyrimido [4,5-b] indol-9-yl)acetate (196b) (311 mg, 0.876 mmol) in DCM (10 mL) using TFA and stirring at RT. This gave after workup 2-(4-amino-6-(methylcarbamoyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (196c) which was used as such in the next step; MS (ES+): 300.10 (M+l); (ES-): 298.05 (M-l).

Step-4: Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5methyl-2-azabicy cio [3.1.0]hexan-2-yl)-2-oxoethyl)-N-methyl-9H-py rimido [4,5 -b] indole-625 carboxamide (196d)

Compound 196d was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(methylcarbamoyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (196c) (131 mg, 0.438 mmol) in DMF (12 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (146 mg, 0.438 mmol), HATU (333 mg, 0.876 mmol) DIPEA (0.381 mL, 2.190 mmol) and stirring at RT for 21 h. This gave after workup and purification by flash column chromatography [silica gel (25 g), eluting with

359

MeOH in DCM from 0 -10%] followed by purification using reverse phase.column chromatography [C18 column eluting with ACN in water (containing 0.1% HCl) from 0100%] 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-N-methyl-9H-pyrimido[4,5-b]indole-65 carboxamide (196d) (19 mg, 7.5% yield) as a white solid; ³H NMR (300 MHz, DMSO-î/ô) δ 10.77 (s, 1H), 9.05 (s, 1H), 8.58 (s, 1H), 8.51 - 8.35 (m, 4H), 8.05 - 7.96 (m, 2H), 7.76 7.65 (m, 2H), 7.31 (d, J = 7.7 Hz, 1H), 5.73 (d, J = 17.4 Hz, 1H), 5.37 (d, J = 17.3 Hz, 1H), 4.37 (dd, J = 9.1, 5.8 Hz, 1H), 3.72 - 3.64 (m, 1H), 2.86 (d, J = 4.4 Hz, 3H), 2.58 - 2.38 (m, 1H), 1.98 (dd, J = 13.2, 5.9 Hz, 1H), 1.30 (s, 3H), 1.09 - 0.97 (m, 1H), 0.96 - 0.90 (m, 1H);

MS (ES+): 577.15 & 579.20 (M+l); MS (ES-): 575.10 & 577.10 (M-l).

Scheme 197

197b

197c

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(o-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (197c)

Step-1: Préparation of ethyl 2-(4-amino-6-(o-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (197a)

360

Compound 197a was prepared according to the procedure reported in step-1 of scheme-59, from ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (150 mg, 0.430 mmol) in dioxane (15 mL) using o-tolylboronic acid (88 mg, 0.644 mmol; CAS # 16419-606), bis(triphenylphosphine)palladium(II) chloride (60.3 mg, 0.086 mmol) a solution of césium 5 carbonate (210 mg, 0.644 mmol) in water (1.8 mL) and heating at 100 °C for 17 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0-5%] ethyl 2-(4-amino-6-(o-tolyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (197a) (70 mg, 45% yield) as an off-white solid; MS (ES+): 361.20 (M+l).

Step-2: Préparation of 2-(4-amino-6-(o-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (197b)

Compound 197b was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-6-(o-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (197a) (64 mg, 0.178 mmol) in THF (3 mL) and methanol (3 mL) using a solution of lithium hydroxide hydrate (45.6 mg, 1.065 mmol) in water (3 mL) and stirring at RT for 14 h. This gave after 15 workup 2-(4-amino-6-(o-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (197b) which was used as such for the next step; MS (ES+): 333.15 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(o-tolyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (197c)

Compound 197c was prepared according to the procedure reported in step-3 of scheme-1, 20 from 2-(4-amino-6-(o-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (197b) (59.2 mg, 0.178 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (85 mg, 0.267 mmol), HATU (169 mg, 0.445 mmol), DIPEA (0.186 mL, 1.068 mmol) and stirring at RT for 19 h. This gave after workup and purification by flash column chromatography [silica gel (25 g), MeOH in DCM from 0 25 10%] followed by purification using reverse phase column chromatography [C18 column, eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino6-(o-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (197c) (23 mg, 22% yield) as a white solid; ^!Η NMR (300 MHz, DMSO-î/ô) δ 10.79 (s, 1H), 8.61 (s, 1H), 8.52 - 8.50 (m, 2H), 8.02 (d, J = 8.2 Hz, 30 1H), 7.78 - 7.66 (m, 2H), 7.52 (dd, J = 8.4,1.5 Hz, 1H), 7.38 - 7.26 (m, 6H), 5.81 (d, J =

17.4 Hz, 1H), 5.45 (d, J = 17.2 Hz, 1H), 4.44 (dd, J = 9.1, 5.5 Hz, 1H), 4.01 - 3.84 (m, 1H),

361

2.46 - 2.14 (m, 5H), 2.02 - 1.83 (m, 1H), 1.17 - 1.03 (m, 1H), 0.89 0.72 (m, 1H); MS (ES+): 596.15 & 598.15; (ES-): 594.10 & 596.10 (M-l).

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (198a)

Compound 198a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)açetic acid (51f) (50 mg, 0.134 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-510 methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (44.4 mg, 0.134 mmol), HATU (76 mg, 0.200 mmol), DIPEA (0.116 mL, 1.068 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 -100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from

0-100%] (lR,3S,5R)-2-(2-(4-amino-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (198a) (48 mg, 67% yield) HCl sait as a white solid;. ^]H NMR (300 MHz, DMSO-ifc) δ 10.78 (s, 1H, D₂O exchangeable), 8.58 (s, 1H), 8.40 (s, 2H, D₂O exchangeable), 8.30 (dd, J = 5.9, 3.0 Hz, 1H), 8.02 (d, J = 8.1 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.38 - 7.29 (m, 3H), 5.75 (d, J = 17.5 Hz,

1H), 5.41 (d, J = 17.5 Hz, 1H), 4.39 (dd, J = 9.0, 6.0 Hz, 1H), 3.66 (dd, J = 5.4, 2.3 Hz, 1H),

2.47 - 2.41 (m, 1H), 1.99 (dd, J = 13.3, 5.9 Hz, 1H), 1.30 (s, 3H), 1.03 (t, J = 5.4 Hz, 1H), 0.86 - 0.71 (m, 1H). ¹⁹F NMR (282 MHz, DMSO) δ -134.67. MS (ES+): 538.1 (M+l); (ES-): 536.1 (M-l); Analysis calculated for C₂₄H₂iBrFN₇O₂.1.75H₂O.HCl: C, 47.54; H, 4.24; Cl, 5.85; N, 16.17; Found: C, 47.63; H, 4.20; Cl, 5.59; N, 15.97.

V 362

Scheme 199

199a

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1,0]hexane-3-carboxamide 5 (199a)

Compound 199a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (16d) (50 mg, 0.118 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicycIo[3.1.0]hexane-3-carboxamide (8a) (39.2 mg, 0.118 10 mmol), HATU (67.2 mg, 0.177 mmol), DIPEA (0.103 mL, 0.589 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-(trifluoromethyl)-9H-pyrimido[4,515 b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (199a) HCl sait (50 mg, 72% yield) HCl sait as a white solid; ¹H NMR (300 MHz, DMSO-dô) δ 10.77 (s, 1H, D₂O exchangeable), 8.84 - 8.69 (m, 3H, 2H D₂O exchangeable), 8.66 (s, 1H), 8.18 (s, 1H), 8.00 (d, J= 8.1 Hz, 1H), 7.79 - 7.61 (m, 2H), 7.31 (d, J= 7.6 Hz, 1H), 5.85 (d, J= 17.4 Hz, 1H), 5.46 (d, J= 17.3 Hz, 1H), 4.38 (dd, J= 9.0, 5.9 20 Hz, 1H), 3.69 (dd, J= 5.6, 2.4 Hz, 1H), 2.48 - 2.40 (m, 1H), 1.99 (dd, J= 13.2, 5.9 Hz, 1H),

1.32 (s, 3H), 1.03 (t, J= 5.4 Hz, 1H), 0.92 (dd, J= 5.4, 2.4 Hz, 1H). ¹⁹F NMR (282 MHz, DMSO) δ -59.34. MS (ES+): 588.1(M+1); (ES-): 586.1 (M-l); Analysis calculated for C₂₅H₂iBrF₃N7O₂.2H₂O.HCl: C, 45.44; H, 3.97; Cl, 5.36; N, 14.84; Found: C, 45.44; H, 3.85; Cl, 5.30; N, 14.74.

Scheme 200

363

200a

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (200a)

Compound 200a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (52e) (60 mg, 0.138 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (45.9 mg, 0.138 mmol), HATU (79 mg, 0.207 mmol), DIPEA (0.120 mL, 0.689 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with 10 DMA-80 in DCM from 0 -100%] followed by purification using reverse phase column chromatography [Cl 8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (200a) (59 mg, 71% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-îZô) δ 10.78 (s, 1H, D2O exchangeable), 8.67 - 8.50 (m, 3H, 2H D₂O exchangeable), 8.46 (d, J= 8.5 Hz, 1H), 8.06 7.95 (m, 2H), 7.70 (t, J= 8.0 Hz, 1H), 7.57 (dd, J= 8.4, 1.7 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.73 (d, J= 17.4 Hz, 1H), 5.36 (d, J= 17.3 Hz, 1H), 4.38 (dd, J= 9.1, 6.0 Hz, 1H), 3.67 (dd, 7=5.6,2.4 Hz, 1H), 2.48-2.42 (m, 1H), 1.98 (dd, J= 13.3, 5.9 Hz, 1H), 1.31 (s, 3H), 1.02 (t, J= 5.6 Hz, 1H), 0.94 (dd, J= 5.3, 2.4 Hz, 1H). MS (ES+): 598.0/600.1 (M+l); (ES-):

596/598.0 (M-l); Analysis calculated for C24H2iBr₂N₇O2 2H₂O.HC1: C, 42.91; H, 3.90; Cl,

5.28; N, 14.60; Found: C, 43.02; H, 3.69; CI, 5.09; N, 14.47.

Scheme 201

364

Préparation of (2S,4R)-4-amino-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (201e)

Step-1: Préparation of (2S,4R)-/er/-butyl 4-((((9H-fluoren-9-yI)methoxy)carbonyl)amino)-2- ((6-bromopyridin-2-yl)carbamoyl)pyrrolidine-1 -carboxylate (201b)

Compound 201b was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4R)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-1 -(tertbutoxycarbonyl)pyrrolidine-2-carboxylic acid (201a) (1 g, 2.210 mmol; CAS # 176486-63-8) in DCM (30 mL) using 1-methyl-lH-imidazole (0.440 mL, 5.52 mmol), methanesulfonyl chloride (0.205 mL, 2.65 mmol) and 6-bromopyridin-2-amine (0.382 g, 2.210 mmol) and stirring at RT for 18 h. This gave after workup (2S,4R)-terZ-butyl 4-((((9H-fluoren-9yl)methoxy)carbonyl)amino)-2-((6-bromopyridin-2-yl)carbamoyl)pyrrolidine-l-carboxylate (201b) (1.22 g, 91% yield); MS (ES+): 607.60 (M+l).

Step-2: Préparation of (9H-fluoren-9-yl)methyl ((3R,5S)-5-((6-bromopyridin-215 yl)carbamoyl)pyrrolidin-3-yl)carbamate (201c)

Compound 201c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-terributyl 4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((6-bromopyridin2-yl)carbamoyl)pyrrolidine-l-carboxylate (201b) (1.2 g, 1.975 mmol) in DCM (11 mL) using TFA (1.065 mL, 13.83 mmol) and stirring ovemight at RT. This gave after workup and

365 purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] HCl sait of (9H-fluoren-9-yl)methyl ((3R,5S)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-3-yl)carbamate (201c) (720 mg, 67% yield); Ή NMR (300 MHz, DMSO-îZ₆) δ 11.47 (s, 1H), 10.15 (s, 1H), 8.94 (s, 1H), 8.06 5 (d, J=8.1Hz, 1H), 7.99 - 7.62 (m, 6H), 7.39 (dt, J= 25.2, 7.3 Hz, 5H), 4.63 -4.51 (m, 1H), ' 4.46-4.31 (m, 2H), 4.31-4.14 (m, 2H), 3.52 (dd, J= 11.7, 6.9 Hz, 1H), 3.14 (dd, J- 12.1,

5.6 Hz, 1H), 2.36 (t, J= 6.7 Hz, 1H), 2.23 (dt, J= 13.9, 7.2 Hz, 1H); MS (ES+): 507.10 (M+l).

Step-3: Préparation of (9H-fluoren-9-yl)methyl ((3R,5S)-l-(2-(4-amino-6-(trifluoromethyl)10 9H-pyrimido[4,5-b] indol-9-yl)acetyl)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-3 yl)carbamate (20Id)

Compound 201d was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (9H-fluoren-9-yl)methyl ((3R,5S)-5-((6-bromopyridin-2yl)carbamoyl)pyrrolidin-3-yl)carbamate (201c) (120 mg, 0.221 mmol) in DMF (3 mL) using 15 TFA sait of 2-(4-amino-6-(trifluoromethyI)-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (lie) (94 mg, 0.221 mmol), HATU (126 mg, 0.331 mmol), DIPEA (0.192 mL, 1.103 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 50%] (9H-fluoren9-yl)methyl ((3R,5S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-920 yl)acetyl)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-3-yl)carbamate (201d) (173 mg, 98% yield) as a white solid; *H NMR (300 MHz, DMSO-dô) δ 10.88 (s, 1H), 8.79 (s, 1H), 8.34 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.92 (d, J= 7.4 Hz, 2H), 7.77 - 7.64 (m, 5H), 7.55 (s, 2H), 7.38 (ddd, J= 27.2, 13.4, 7.5 Hz, 6H), 5.34 (s, 2H), 4.55 (s, 1H), 4.43 (d, J= 6.7 Hz, 2H), 4.28 (d, J= 6.7 Hz, 1H), 4.17 (d, J= 9.2 Hz, 1H), 3.61 (s, 1H), 3.22 - 3.02 (m, 0H),

2.24 (d, J= 19.2 Hz, 1H), 2.09 (d, J= 6.0 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -58.18.

Step-4: Préparation of (2S,4R)-4-amino-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (201e)

Compound 201e was prepared according to the procedure reported in step-4 of scheme-180, from (9H-fluoren-9-yl)methyl ((3R,5S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,530 b]indol-9-yl)acetyl)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-3-yl)carbamate (201d) (170 mg, 0.213 mmol) in DMF (0.8 mL) using piperidine (0.210 mL, 2.126 mmol) and stirring at RT for 1 h. This gave after work up and purification using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 -100%] followed

366 by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with

ACN m water (containing 0.1% HCl) from 0-100%] (2S,4R)-4-amino-l-(2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)pyrrolidine2-carboxamide (201e) (59 mg, 48% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/ô) δ (a mixture oftwo rotamers) 11.50 and 11.04 (2s, 1H, D₂O exchangeable), 9.02 (s, 1H), 8.91 (s, 2H, D₂O exchangeable), 8.69 (m, 4H, 3H D2O exchangeable), 8.21 - 7.96 (m, 2H), 7.89 - 7.81 (m, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.43 and 7.34 (2d, J= 7.7 Hz, 1H), 5.63-5.49 (m, 2H), 4.70 (dd, J= 8.5, 6.0 Hz, 1H), 4.21 (dd, J= 11.2, 6.2 Hz, 1H), 4.13 - 3.97 (m, 2H), 2.48 (m, 1H), 2.27 (dt, J= 13.1, 6.1 Hz, 1H). ¹⁹F NMR (282 MHz, DMSO-î7₆) δ 58.58. MS (ES+): 577.1 (M+l); (ES-): 575.1 (M-l); Analysis calculated for CzsHzoBrFsNsCh 1.25H₂O.1.9HC1.0.5DMSO: C, 40.70; H, 3.90; Cl, 9.51; N, 15.82; Found: C, 40.41; H, 3.65; Cl, 9.32; N, 15.85.

Scheme 202

188f

202a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (202a)

Compound 202a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (188f) (91 mg, 0.317 mmol) in DMF (12 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (8a) (105 mg, 0.317 mmol), HATU (241 mg, 0.634 mmol) and DIPEA (0.276 mL, 1.585 mmol) and stirring at RT for 13 h. This gave after workup and purification using reverse phase column chromatography [Cl8 column, eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-nitro9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (202a) (42 mg, 23% yield) as a white solid; ’H NMR

367 (300 MHz, DMSO-Jô) δ 10.77 (s, 1H), 9.52 (d, J= 2.2 Hz, 1H), 8.72 (s, 2H), 8.63 (s, 1H),

8.40 (dd, J= 9.1, 2.2 Hz, 1H), 8.00 (d, J= 8.2 Hz, 1H), 7.87 (d, J= 9.1 Hz, 1H), 7.69 (t, J= 8.0 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.79 (d, J= 17.4 Hz, 1H), 5.43 (d, J= 17.3 Hz, 1H), 4.37 (dd, J= 9.0, 6.0 Hz, 1H), 3.68 (dd, J= 5.6, 2.4 Hz, 1H), 2.50-2.42 (m, 1H), 1.98 (dd, J = 13.2, 5.9 Hz, 1H), 1.31 (s, 3H), 1.02 (t, J= 5.4 Hz, 1H), 0.95 (dd, J= 5.4, 2.4 Hz, 1H); MS (ES+): 565.10 & 567.10 (M +1); MS (ES-): 563.10 & 565.00 (M -1); Analysis calculated for C24H2iBrN₈04.0.85HCL2.0H₂0: C, 45.58; H, 4.12; N, 17.72; Cl, 4.77; Found: C, 45.65; H, 3.97; N, 17.42.; Cl, 4.59.

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-6-carboxamide (203a)

Compound 203a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-carbamoyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (60b) (0.079 g, 0.278 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (111 mg, 0.334 mmol), HATU (211 mg, 0.556 mmol), DIPEA (0.242 mL, 1.39 mmol) and stirring at RT for 22 h. This gave after workup and purification using reverse phase column chromatography [Cl 8 column , eluting with ACN in water (containing 0.1% HCl) from 0-100%] 4-amino-9-(2-((lR,3S,5R)-3-((6bromopyridin-2-yI)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9Hpyrimido[4,5-b]indole-6-carboxamide (203a) (42 mg, 27% yield) as a white solid; Ή NMR (300 MHz, DMSO-ί/ό) δ 10.77 (s, 1H), 9.14 (d, J= 1.6 Hz, 1H), 8.66 (s, 3H), 8.62 (s, 1H), 8.07 (dd, J= 8.7, 1.5 Hz, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.96 (s, 1H), 7.76 - 7.64 (m, 2H), 7.48 (s, 1H), 7.31 (d,J=7.7Hz, 1H), 5.75 (d, J= 17.4 Hz, 1H), 5.38 (d, J= 17.3 Hz, 1H), 4.38 (dd, J= 9.1, 5.9 Hz, 1H), 3.77 - 3.63 (m, 1H), 2.53-2.38 (m, 1H), 1.98 (dd, J= 13.1,

368

5.8 Hz, 1H), 1.30 (s, 3H), 1.07 - 0.97 (m, 1H), 0.96 - 0.89 (m, 1H); MS (ES+): 563.20 &

565.10 (M+l); MS (ES-): 561.10 & 563.10 (M-l); Analysis calculated for C2₅H23BrN8O3.1.0HC1.2.75H₂O: C, 46.24; H, 4.58; N, 17.25; Found: C, 46.38; H, 4.51; N, 16.99.

Scheme 204

196c

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicycIo[3.1.0]hexan-2-yl)-2-oxoethyl)-N-methyl-9H-pyrimido[4,5-b]indole-6carboxamide (204a)

Compound 204a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(methylcarbamoyI)-9H-pyrimido[4,5-b]indol-9-yI)acetic acid (196c) (0.131 g, 0.438 mmol) in DMF (12 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (167 mg, 0.526 mmol), HATU (333 mg, 0.876 mmol), DIPEA (0.381 mL, 2.19 mmol) and stirring at RT for 21 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with MeOH in DCM from 0 - 8% then 15%] followed by purification using reverse phase column chromatography [Cl8 column, eluting with ACN in water (containing 0.1% HCl) from 0100%] 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yI)-2-oxoethyl)-N-methyl-9H-pyrimido[4,5-b]indole-620 carboxamide (204a) (12 mg, 5% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-Je) δ 10.77 (s, 1H), 9.13 (s, 1H), 8.66 (s, 2H), 8.62 (s, 1H), 8.55 - 8.44 (m, 1H), 8.06 - 7.95 (m, 2H), 7.78 - 7.66 (m, 2H), 7.31 (d, J= 7.7 Hz, 1H), 5.79 (d, J= 17.3 Hz, 1H), 5.43 (d, J= 17.3 Hz, 1H), 4.42 (dd, J= 9.1, 5.4 Hz, 1H), 3.98 - 3.83 (m, 1H), 2.86 (d, J= 4.4 Hz, 3H), 2.41-2.14 (m, 2H), 2.01-1.82 (m, 1H), 1.17-0.94 (m, 1H), 0.87-0.60 (m, 1H); MS (ES+): 563.15 & 565.10 (M+l), MS (ES-): 561.10 & 563.05 (M-l).

369

Scheme 205

205b

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(p-toIyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyI)N-(6-bromopyridin-2-yI)-2-azabicyclo [3.1.0]hexane-3 -carboxamide (205c)

Step-1 : Préparation of ethyl 2-(4-amino-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (205a)

Compound 205a was prepared according to the procedure reported in step-1 of scheme-62, from ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (200 mg, 0.573 mmol) in dioxane (18 mL) usingp-tolylboronic acid (117 mg, 0.859 mmol; CAS # 16419-6010 6), bis(triphenylphosphine)palladium(II) chloride (80 mg, 0.115 mmol) a solution of césium carbonate (280 mg, 0.859 mmol) in water (2.2 mL) and heating at 100 °C for 13 h. This gave after workup and purification by flash column chromatography [silica gel (25 g), eluting with MeOH in DCM from 0-5%] ethyl 2-(4-amino-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (205a) (52 mg, 25% yield) as an off-white solid; 1H NMR (300 MHz, DMSO-J6) δ 8.62 (d, J= 1.6 Hz, 1H), 8.29 (s, 1H), 7.76 (d, J= 8.0 Hz, 2H), 7.71 (dd, J= 8.6, 1.6 Hz, 1H), 7.65 (d, J= 8.5 Hz, 1H), 7.44 (s, 2H), 7.29 (d, J= 7.9 Hz, 2H), 5.26 (s, 2H), 4.15 (q, J = 7.1 Hz, 2H), 2.36 (s, 3H), 1.21 (t, J= 7.1 Hz, 3H); MS (ES+): 361.20 (M+l).

370

Step-2: Préparation of 2-(4-amino-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (205b)

Compound 205b was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-6-(p-tolyl)-9H-pyrimido[4,5-b]indoI-9-yI)acetate (205a) (50 mg, 5 0.139 mmol) in THF (3 mL) and methanol (3 mL) using a solution of lithium hydroxide hydrate (35.6 mg, 0.832 mmol) in water (3 mL) and stirring at RT for 18 h. This gave after workup 2-(4-amino-6-(p-tolyI)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (205b) which was used as such for the next step; MS (ES+): 333.10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-910 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (205c)

Compound 205c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (205b) (46 mg, 0.139 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (66.4 mg, 0.209 mmol), HATU (106 mg, 0.278 15 mmol), DIPEA (0.121 mL, 0.695 mmol) and stirring at RT for 14 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [C 18 column, eluting with ACN in water (containing 0.1% HCl) from 0-100%] (1R,3S,5R)2-(2-(4-amino-6-(p-toIyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-220 azabicyclo[3.1.0]hexane-3-carboxamide (205c) (42 mg, 51% yield) HCl sait as a white solid;

Ή NMR (300 MHz, DMSO-cfe) δ 10.77 (s, 1H), 8.77 (d, J= 1.6 Hz, 1H), 8.70 (s, 2H), 8.62 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.86 (dd, J= 8.6, 1.6 Hz, 1H), 7.78 - 7.65 (m, 4H), 7.34 7.29 (m, 3H), 5.78 (d, J= 17.3 Hz, 1H), 5.44 (d, J= 17.3 Hz, 1H), 4.42 (dd, J= 9.0, 5.5 Hz, 1H), 4.01 - 3.84 (m, 1H), 2.40 - 2.29 (m, 4H), 2.28 - 2.13 (m, 1H), 2.01 - 1.85 (m, 1H), 1.16 25 - 1.02 (m, 1H), 0.84 - 0.73 (m, 1H); MS (ES+): 596.20 & 598.10 (M+l), MS (ES-): 594.10 & 596.10 (M-l); Analysis calculated for C3oH26BrN702.HC1.2.25H20: C, 53.50; H, 4.71; N, 14.56; Cl, 5.26; Found: C, 53.68; H, 4.56; N, 14.58.; Cl, 5.07.

Scheme 206

371

206b '

206c

Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyI)-N-(6-bromopyridin-2-yl)-4-methylpyrrolidine-2-carboxamide (206c)

Step-1: Préparation of (2S,4R)-terZ-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-45 methylpyrrolidine-l-carboxylate (206a)

Compound 206a was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4R)-l-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic acid (121a) (0.5 g, 2.181 mmol) in DCM (15 mL) using 1-methyl-lH-imidazole (0.435 mL, 5.45 mmol), methanesulfonyl chloride (0.203 mL, 2.62 mmol), 6-bromopyridin-2-amine (0.377 g, 2.181 10 mmol) and stirring at RT for 18 h. This gave after workup (2S,4R)-Zer/-butyl 2-((6bromopyridin-2-yl)carbamoyl)-4-methylpyrrolidine-l-carboxylate (206a) (656 mg, 78% yield); ’HNMR (300 MHz, DMSO-î/₆) δ 10.90 (d, J= 12.5 Hz, 1H), 8.07 (dd, J= 11.8, 8.1 Hz, 1H), 7.74 (q, J= 7.6 Hz, 1H), 7.34 (d, J= 7.7 Hz, 1H), 4.50 - 4.30 (m, 1H), 3.58 (dt, J= 10.0, 6.7 Hz, 1H), 2.91 - 2.79 (m, 1H), 2.33 (dt, J= 15.6, 7.4 Hz, 1H), 1.97 (q, J= 4.4 Hz, 15 1H), 1.89 - 1.72 (m, 1H), 1.32 (d, J= 39.6 Hz, 9H), 0.96 (d, J= 6.6 Hz, 3H); MS (ES+):

284.00 (M-Boc+1).

Step-2: Préparation of (2S,4R)-N-(6-bromopyridin-2-yl)-4-methylpyrrolidine-2-carboxamide (206b)

372

Compound 206b was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-teri-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-4-methylpyrrolidine-lcarboxylate (206a) (650 mg, 1.692 mmol) in DCM (9 mL) using TFA (0.912 mL, 11.84 mmol) and stirring ovemight at RT. This gave after workup and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] HCl sait of (2S,4R)-N-(6-bromopyridin-2-yl)-4-methylpyrrolidine2-carboxamide (206b) (541 mg, 100% yield) as a white solid; *H NMR (300 MHz, DMSOd₆) δ 11.44 (s, 1H), 9.63 (s, 1H), 8.68 (s, 1H), 8.05 (d, J= 8.2 Hz, 1H), 7.82 (t, J= 7.9 Hz, 1H), 7.44 (d, J= Ί.Ί Hz, 1H), 4.49 (s, 1H), 3.45 (s, 1H), 2.80 (s, 1H), 2.32 (dd, J= 14.9, 7.6 Hz, 1H), 2.19 (dt, J= 12.5, 6.0 Hz, 1H), 1.98 (dt, J= 13.1, 8.7 Hz, 1H), 1.03 (d, J= 6.6 Hz, 3H).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-methylpyrrolidine-2-carboxamide (206c)

Compound 206c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (2S,4R)-N-(6-bromopyridin-2-yl)-4-methylpyrrolidine-2-carboxamide (206b) (50 mg, 0.156 mmol) in DMF (1.5 mL) using TFA sait of 2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (66.2 mg, 0.156 mmol), HATU (89 mg, 0.234 mmol), DIPEA (0.136 mL, 0.780 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 50%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-4-methylpyrrolidine-2-carboxamide (206c) (41 mg, 46% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-àfe) (a mixture of two rotamers) δ 11.22 and 10.78 (2s, 1H, D₂O exchangeable), 8.94 (s, 1H), 8.81 (s, 2H, D₂O exchangeable), 8.61 and 8.58 (2s, 1H), 8.10 and 7.93 (2d, J= 8.1 Hz, 1H), 7.89 - 7.72 (m, 2H), 7.63 (t, J= 8.0 Hz, 1H), 7.33 and 7.24 (d, J= 7.7 Hz, 1H), 5.43 (q, J= 17.3 Hz, 2H), 4.60 - 4.43 (m, 1H), 4.03 (t, J= 8.5 Hz, 1H), 3.34 (t, 1H), 2.54 - 2.46 (m, 1H), 2.00 (dd, J= 13.2, 6.6 Hz, 1H), 1.88 - 1.70 (m, 1H), 1.04 and 0.96 (2d, J= 6.5 Hz, 3H). ¹⁹F NMR (282 MHz, DMSO) δ -58.59. MS (ES+): 576.1 (M+l); (ES-): 574.1 (M-l); Analysis calculated for C₂₄H₂iBrF₃N₇O₂ 1.5H₂O.HC1: C, 45.05; H, 3.94; Cl, 5.54; N, 15.32; Found: C, 45.09; H, 3.91; Cl, 5.42; N, 15.09.

Scheme 207

373

CF₃ 11e 207b

Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (207b)

Compound 207b was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (60 mg, 0.141 mmol) in DMF (1.2 mL) using TFA sait of (2S,4R)-N-(6-bromopyridin2-yl)-4-fhioro-4-methylpyrrolidine-2-carboxamide (207a) (58.9 mg, 0.141 mmol; prepared according to the procedure reported in Wiles, Jason A. et al., PCT Int. Appl. (2017), WO 2017035353 Al 20170302), HATU (81 mg, 0.212 mmol), DIPEA (0.123 mL, 0.707 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 50%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoro-415 methylpyrrolidine-2-carboxamide (207b) (55 mg, 65% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-àfe) (a mixture of two rotamers) δ 11.35 and 10.96 (2s, 1H, D₂O exchangeable), 8.95 (s, 1H), 8.57 (s, 1H), 8.47 (s, 2H, D₂O exchangeable), 8.21 and 7.98 (2d, J= 8.2 Hz, 1H), 7.84 (s, 2H), 7.68 (t, J= 8.0 Hz, 1H), 7.43 and 7.31 (2d, J= 7.7 Hz, 1H), 5.61 (d, J= 17.4 Hz, 1H), 5.38 (d, J= 17.3 Hz, 1H), 4.62 (t, 8.6 Hz, 1H), 4.30 (dd, J=

18.9, 12.0 Hz, 1H), 3.97 (dd, J= 35.2, 11.9 Hz, 1H), 2.54 (m, 1H), 2.20 - 1.93 (m, 1H), 1.62 (d, J= 21.0 Hz, 3H). ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -58.50, -139.31. MS (ES+): 594.1 (M+l); (ES-): 592.0 (M-l); Analysis calculated for C₂4H₂₀BrF₄N₇O₂ 1.25H₂O.HC1.: C, 44.12; H, 3.63; Cl, 5.43; N, 15.01; Found: C, 44.16; H, 3.67; Cl, 5.59; N, 14.84.

Scheme 208

374

208c

Préparation of (2S,4S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-4-methylpyrrolidine-2-carboxamide (208d)

Step-1: Préparation of (2S,4S)-ZerZ-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-45 methylpyrrolidine-l-carboxylate (208b)

Compound 208b was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4S)-l-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic acid (208a) (0.45 g, 1.963 mmol; CAS # 364750-81-2) in DCM (15 mL) using 1-methyl-IH-imidazole (0.391 mL, 4.91 mmol), methanesulfonyl chloride (0.182 mL, 2.355 mmol), 6-bromopyridin-210 amine (0.340 g, 1.963 mmol) and stirring at RT for 18 h. This gave after workup (2S,4S)-ZerZbutyl 2-((6-bromopyridin-2-yl)carbamoyl)-4-methylpyrrolidine-l-carboxylate (208b) (623 mg, 83% yield); 'HNMR (300 MHz, DMSO-z7₆) δ 10.92 (d, J= 16.6 Hz, 1H), 8.10 (dd, J= 14.1, 8.2 Hz, 1H), 7.76 (t, J= 8.1 Hz, 1H), 7.34 (d,J=7.7Hz, 1H), 4.32 (t, J= 8.3 Hz, 1H), 3.70-3.49 (m, 1H), 2.93 - 2.79 (m, 1H), 2.40-2.10 (m, 2H), 1.30 (d,J=43.9 Hz, 10H),

1.00 (t, J= 6.1 Hz, 3H); MS (ES+): 384.10 (M+l).

Step-2: Préparation of (2S,4S)-N-(6-bromopyridin-2-yl)-4-methylpyrrolidine-2-carboxamide (208c)

Compound 208c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4S)-ZerZ-butyl 2-((6-bromopyridin-2-yl)carbamoyI)-4-methylpyrrolidine-1 20 carboxylate (208b) (610 mg, 1.587 mmol) in DCM (9 mL) using TFA (0.856 mL, 11.11

375 mmol) and stirring ovemight at RT. This gave after workup TFA sait of (2S,4S)-N-(6bromopyridin-2-yl)-4-methylpyrrolidine-2-carboxamide (208c) (621 mg, 98% yield); *H NMR (300 MHz, DMSO-î/ô) δ 11.41 (s, 1H), 9.31 (s, 1H), 8.75 (s, 1H), 8.07 (d, J= 8.2 Hz, 1H), 7.82 (t, J= 8.0 Hz, 1H), 7.44 (d, J= 7.7 Hz, 1H), 4.51 - 4.27 (m, 1H), 3.54 - 3.26 (m,

1H), 2.91-2.64 (m, 1H), 2.59 (dd, J= 13.2, 7.1 Hz, 1H), 2.38-2.14 (m, 1H), 1.53 (dt, J=

12.6, 10.0 Hz, 1H), 1.04 (d, J= 6.6 Hz, 3H); MS (ES+): 284.00 (M+l).

Step-3: Préparation of (2S,4S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-4-methylpyrrolidine-2-carboxamide (208d)

Compound 208d was prepared according to the procedure reported in step-2 of scheme-1, from TFA sait of (2S,4S)-N-(6-bromopyridin-2-yl)-4-methylpyrrolidine-2-carboxamide (208c) (56.3 mg, 0.141 mmol) in DMF (1.2 mL) using TFA sait of 2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (60 mg, 0.141 mmol) HATU (81 mg, 0.212 mmol), N-ethyl-N-isopropylpropan-2-amine (0.123 mL, 0.707 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 50%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4S)-l-(2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yI)-4methylpyrrolidine-2-carboxamide (208d) (60 mg, 74% yield) HCl sait as a white solid; ’H

NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 11.31 and 10.86 (2s, 1H, D₂O exchangeable), 8.97 (s, 1H), 8.61 (s, 3H, 2H D₂O exchangeable), 8.20 and 7.99 (2d, J= 8.2 Hz, 1H), 7.92 - 7.81 (m, 2H), 7.68 (t, J= 8.0 Hz, 1H), 7.41 and 7.30 (2d, J= 7.7 Hz, 1H), 5.55 (d, J= 17.3 Hz, 1H), 5.38 (d, J= 17.3 Hz, 1H), 4.44 (t, J= 8.1 Hz, 1H), 4.14 (t, 8.4

Hz, 1H), 3.34 (t, J= 9.6 Hz, 1H), 2.54 (m, 1H), 2.42 (t, J= 7.2 Hz, 1H), 1.49 (q, J= 9.6 Hz,

1H), 1.13 andl.02 (2d, J= 6.6 Hz, 3H). ¹⁹F NMR (282 MHz, DMSO-î/ô) δ -58.53. MS (ES+):

576.1 (M+l); (ES-): 574.0 (M-l); Analysis calculated for C₂₄H₂iBrF₃N₇O₂ 1.5H₂O.HCL: C, 45.05; H, 3.94; Cl, 5.54; N, 15.32; Found: C, 45.08; H, 4.01; Cl, 5.43; N, 15.12.

Scheme 209

Préparation of (2S,4R)-1 -(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (209b)

Compound 209b was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (60 mg, 0.141 mmol) in DMF (1.2 mL) using HCl sait of (2S,4R)-N-(6-bromo-3methylpyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (209a) (50 mg, 0.141 mmol; prepared according to the procedure reported in Wiles, Jason A. et al., PCT Int. Appl.

(2018), WO 2018160889 Al 20180907.), HATU (81 mg, 0.212 mmol) DIPEA (0.123 mL,

0.707 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 50%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-615 (trifluoromethyl)-9H-pyrimido[4,5-b] indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-4fluoro-4-methylpyrrolidine-2-carboxamide (209b) (33 mg, 38% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-Jô) (a mixture of two rotamers) δ 10.83 and 10.40 (2s, 1H, D₂O exchangeable), 8.94 (s, 1H), 8.69 (s, 2H, D₂O exchangeable), 8.56 (s, 1H), 7.76 (qd, J = 10.7, 9.7, 6.2 Hz, 2H), 7.66 and 7.51 (d, J= 8.0 Hz, 1H), 7.46 and 7.35 (2d, J= 7.9 Hz, 1H),

5.58 (d, J= 17.3 Hz, 1H), 5.31 (d, J= 17.3 Hz, 1H), 4.51 (dd, J= 9.8, 7.5 Hz, 1H), 4.24 (dd,

J= 18.9, 12.3 Hz, 1H), 3.89 (dd, J= 35.1, 12.0 Hz, 1H), 2.56 (td, J= 14.9, 7.6 Hz, 1H), 2.18 - 1.93 (m, 1H), 1.85 (s, 3H), 1.58 (d, J= 21.1 Hz, 3H). ¹⁹F NMR (282 MHz, DMSO-ifc) δ 58.56, -139.44; MS (ES+): 608.1 (M+l), 630.1 (M+Na); (ES-): 606.1 (M-l); Analysis calculated for C₂₅H₂₂BrF4N₇O₂ 1.25H₂O.HC1.: C, 44.99; H, 3.85; N, 14.69; Found: C, 45.03;

H, 3.79; N, 14.43.

Scheme 210

377

210c

210d

Préparation of (S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-4,4-dimethylpyrrolidine-2-carboxamide (210d)

Step-1: Préparation of (S)-teri-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-4,45 dimethylpyrrolidine-l-carboxylate (210b)

Compound 210b was prepared according to the procedure reported in step-1 of scheme-53, from (S)-l-(tert-butoxycarbonyl)-4,4-dimethylpyrrolidine-2-carboxyIic acid (210a) (0.45 g, 1.850 mmol; CAS # 87691-27-8) in DCM (15 mL) using 1-methyl-1 H-imidazole (0.369 mL, 4.62 mmol), methanesulfonyl chloride (0.172 mL, 2.219 mmol), 6-bromopyridin-2-amine (0.320 g, 1.850 mmol) and stirring at RT for 18 h. This gave after workup (S)-fôrributyl 2((6-bromopyridin-2-yl)carbamoyl)-4,4-dimethylpyrrolidine-l-carboxylate (210b) (570 mg, 77% yield); *HNMR (300 MHz, DMSO-î/₆) δ 10.89 (d, J= 19.4 Hz, 1H), 8.10 (dd, J= 14.6, 8.2 Hz, 1H), 7.75 (q, J= 8.3 Hz, 1H), 7.34 (d, J= 7.7 Hz, 1H), 4.53 - 4.32 (m, 1H), 3.22 (dd, J= 10.2, 5.8 Hz, 1H), 3.09 (dd, J= 10.2, 5.2 Hz, 1H), 2.01 (dt, J= 12.5, 6.5 Hz, 1H), 1.70 15 1.55 (m, 1H), 1.31 (d, J= 40.7 Hz, 9H), 1.07 (d, J= 5.3 Hz, 3H), 1.00 (s, 3H); MS (ES+):

398.10 (M+l).

Step-2: Préparation of (S)-N-(6-bromopyridin-2-yl)-4,4-dimethylpyrrolidine-2-carboxamide (210c)

378

Compound 210c was prepared according to the procedure reported in step-2 of scheme-1, from (S)-tert-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-4,4-dimethylpyrrolidine-1 carboxylate (210b) (570 mg, 1.431 mmol) in DCM (8 mL) using TFA (0.772 mL, 10.02 mmol) and stirring ovemight at RT. This gave after workup and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] to afford the HCl sait of (S)-N-(6-bromopyridin-2-yl)-4,4dimethylpyrrolidine-2-carboxamide (210c) (375 mg, 78% yield); 'H NMR (300 MHz, DMSO-î/6) δ 11.44 (s, 1H), 10.24 (s, 1H), 8.81 (s, 1H), 8.06 (d, J= 8.2 Hz, 1H), 7.83 (t, J= 7.9 Hz, 1H), 7.44 (d, J= 7.7 Hz, 1H), 4.52 (s, 1H), 3.04 (q, J= 5.6, 5.1 Hz, 2H), 2.30 (dd, J=

12.8, 8.3 Hz, 1H), 1.80 (dd, J= 12.9, 9.4 Hz, 1H), 1.11 (d, J= 2.5 Hz, 6H); MS (ES+):

298.10 (M+l).

Step-3: Préparation of (S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-4,4-dimethylpyrrolidine-2-carboxamide (210d)

Compound 210d was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (S)-N-(6-bromopyridin-2-yl)-4,4-dimethylpyrrolidine-2-carboxamide (210c) (47.3 mg, 0.141 mmol) in DMF (1.2 mL) using TFA sait of 2-(4-amino-6-(trifluoromethyl)9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (60 mg, 0.141 mmol), HATU (81 mg, 0.212 mmol), DIPEA (0.123 mL, 0.707 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in

DCM from 0 - 50%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (S)-l(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2yl)-4,4-dimethylpyrrolidine-2-carboxamide (210d) (34 mg, 41% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) (a mixture oftwo rotamers) δ 11.26 and 10.83 (2s, 1H,

D2O exchangeable), 8.92 (s, 1H), 8.53 and 8.49 (2s, 1H), 8.26 (s, 2H, D₂O exchangeable), 8.20 and 7.99 (2d, J= 8.2 Hz, 1H), 7.81 (s, 2H), 7.68 (t, J- 8.0 Hz, 1H), 7.41 and 7.31 (2d, J = 7.7 Hz, 1H), 5.52 (d, J= 17.3 Hz, 1H), 5.34 (d, J= 17.3 Hz, 1H), 4.54 (t, J= 8.3 Hz, 1H), 3.77 (d, J= 9.8 Hz, 1H), 3.60 (d, J= 9.9 Hz, 1H), 2.08 (dd, J= 12.4, 7.9 Hz, 1H), 1.71 (dd, J = 12.4, 9.0 Hz, 1H), 1.21 and 1.10 (2s, 3H), 1.14 and 1.08 (2s, 3H). ¹⁹FNMR(282 MHz,

DMSO-îZô) δ -58.42. MS (ES+): 590.0 (M+l), 612.0 (M+Na); (ES-): 588.0 (M-l); Analysis calculated for C25H23BrF₃N7O₂ 1.25H₂O.HC1: C, 46.24; H, 4.11; Cl, 5.46; N, 15.10; Found: C, 46.21; H, 4.05; Cl, 5.62; N, 14.78.

Scheme 211

379

211d

Préparation of (2S,4S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-4-(aminomethyI)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (211e)

Step-1: Préparation of (2S,4S)-/erZ-butyl 4-(((((9H-fluoren-95 yl)methoxy)carbonyl)amino)methyl)-2-((6-bromopyridin-2-yl)carbamoyl)pyrrolidine-1 carboxylate (211b)

Compound 211b was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4S)-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-1 -(tertbutoxycarbonyl)pyrrolidine-2-carboxylic acid (211a) (0.25 g, 0.536 mmol; CAS # 27322110 98-0) in DCM (10 mL) using 1-methyl-lH-imidazole (0.107 mL, 1.340 mmol), methanesulfonyl chloride (0.050 mL, 0.643 mmol), 6-bromopyridin-2-amine (0.093 g, 0.536 mmol) and stirring at RT for 18 h. This gave after workup (2S,4S)-ter/-butyl 4-(((((9Hfluoren-9-yl)methoxy)carbonyl)amino)methyl)-2-((6-bromopyridin-2yl)carbamoyl)pyrrolidine-l-carboxylate (211b) (67 mg, 20% yield); MS (ES+): 621.20 (M+l).

Step-2: Préparation of (9H-fluoren-9-yl)methyl (((3R,5S)-5-((6-bromopyridin-2yl)carbamoyl)pyrrolidin-3-yl)methyl)carbamate (211c)

380

Compound 211c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4S)-terZ-butyl 4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-2-((6bromopyridin-2-yl)carbamoyl)pyrrolidine-l-carboxylate (211b) (65 mg, 0.105 mmol) in DCM (1 mL) using TFA (0.056 mL, 0.732 mmol) and stirring ovemight at RT. This gave after workup TFA sait of (9H-fluoren-9-yl)methyl (((3R,5S)-5-((6-bromopyridin-2yl)carbamoyl)pyrrolidin-3-yI)methyl)carbamate (211c) (60 mg, 90% yield); *HNMR (300 MHz, DMSO-ίΖό) δ 11.44 (s, 1H), 9.38 (s, 1H), 8.75 (s, 1H), 8.06 (d, J= 8.2 Hz, 1H), 7.87 (dd, J= 15.0, 7.7 Hz, 3H), 7.67 (d, J= 7 A Hz, 2H), 7.51 - 7.26 (m, 6H), 4.47 (s, 1H), 4.36 (d, J= 6.8 Hz, 2H), 4.22 (t, J= 6.7 Hz, 1H), 3.42 (s, 1H), 3.02 (dt, J= 33.5, 7.? Hz, 3H), 2.41 (t, J= 7.6 Hz, 1H), 2.10 (t, J= 8.9 Hz, 2H).

Step-3: Préparation of (9H-fluoren-9-yl)methyl (((3S,5S)-l-(2-(4-amino-6-(trifluoromethyI)9H-pyrimido[4,5-b]indol-9-yI)acetyl)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-3yl)methyl)carbamate (21 Id)

Compound 211d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (9H-fluoren-9-yl)methyl (((3R,5S)-5-((6-bromopyridin-2yl)carbamoyl)pyrrolidin-3-yl)methyl)carbamate (211c) (55 mg, 0.087 mmol) in DMF (1 mL) using TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (36.7 mg, 0.087 mmol), HATU (49.4 mg, 0.130 mmol), DIPEA (0.075 mL, 0.433 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 50%] (9H-fluoren9-yl)methyl (((3S,5S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-5 -((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-3 -yl)methyl)carbamate (211 d) (70 mg, 99% yield) as a white solid; Ή NMR (300 MHz, DMSO-î7₆) δ 10.87 (s, 1H), 8.80 (s, 1H), 8.33 (s, 1H), 8.02 (d, 8.2 Hz, 1H), 7.88 (d, J= 7.4 Hz, 2H), 7.74 - 7.66 (m, 5H),

7.55 (s, 3H), 7.44 - 7.26 (m, 5H), 5.34 (q, J= 17.3 Hz, 2H), 4.57 (s, 1H), 4.39 (d, J= 6.7 Hz,

2H), 4.25 (t, J= 6.7 Hz, 1H), 4.00 (t, J= 8.6 Hz, 1H), 3.71 - 3.44 (m, 1H), 3.21 - 3.04 (m, 3H), 1.98 (d, J= 7.8 Hz, 2H), 1.22 (s, 1H); MS (ES+): 813.10 (M+l).

Step-4: Préparation of (2S,4S)-l-(2-(4-amino-6-(trifluoromethyI)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-4-(aminomethyl)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (211e)

Compound 211e was prepared according to the procedure reported in step-4 of scheme-180, from (9H-fluoren-9-yl)methyl (((3 S,5 S)-1 -(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido [4,5 b]indol-9-yl)acetyl)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-3-yl)methyl)carbamate (211d) (60 mg, 0.074 mmol) in DMF (0.4 mL) using piperidine (0.073 mL, 0.737 mmol) and

381 ” stirring at RT for 1 h. This gave after work up and purification using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4S)-l-(2-(4-amino-65 (trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-4-(aminomethyl)-N-(6bromopyridin-2-yl)pyrrolidine-2-carboxamide (211e) (21 mg, 48% yield) HCl sait as a white solid; ^XH NMR (300 MHz, DMSOX) (a mixture of two rotamers) δ 11.40 and 10.93 (2s, 1H, D₂O exchangeable), 9.00 (s, 1H), 8.79 (s, 2H, D2O exchangeable), 8.66 and 8.62 (2s, 1H), 8.26 (s, 3H, D₂O exchangeable), 8.02 - 7.78 (m, 3H), 7.71 (t, J= 8.0 Hz, 1H), 7.42 and 7.33 (2d, J= 7.6 Hz, 1H), 5.53 (s, 2H), 4.63 (d, J= 8.3 Hz, 1H), 4.13 (dd, J= 9.9, 7.2 Hz, 1H),

3.73 (t, J= 8.8 Hz, 1H), 3.05 - 2.88 (m, 2H), 2.84 - 2.69 (m, 1H), 2.28 - 1.99 (m, 2H). ^I9F NMR (282 MHz, DMSOX) δ -58.56. MS (ES+): 591.1(M+1); (ES-): 589.1 (M-l).

Scheme 212

212a

TFA

212b

212d

212e

Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-4-(aminomethyl)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (212e)

Step-1: Préparation of (2S,4R)-Zer/-butyl 4-(((((9H-fluoren-9yl)methoxy)carbonyl)amino)methyl)-2-((6-bromopyridin-2-yl)carbamoyl)pyrrolidine-lcarboxylate (212b)

382

Compound 212b was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4R)-4-(((((9H-fluoren-9-yl)rnethoxy)carbonyl)amino)methyl)-l-(/erA butoxycarbonyl)pyrrolidine-2-carboxylic acid (212a) (0.25 g, 0.536 mmol; CAS # 217305284-9) in DCM (10 mL) using 1-methyl-IH-imidazole (0.107 mL, 1.340 mmol), methanesulfonyl chloride (0.050 mL, 0.643 mmol), 6-bromopyridin-2-amine (0.093 g, 0.536 mmol) and stirring at RT for 18 h. This gave after workup (2S,4R)-tert-butyl 4-(((((9Hfluoren-9-yl)methoxy)carbonyl)amino)methyl)-2-((6-bromopyridin-2yl)carbamoyl)pyrrolidine-l-carboxylate (212b) (231 mg, 69% yield); 'H NMR (300 MHz, DMSO-î/ô) δ 10.93 (d, J= 15.8 Hz, 1H), 8.11 (dd, J= 14.0, 8.1 Hz, 1H), 7.89 (d, J= 7.4 Hz, 10 2H), 7.81 - 7.60 (m, 3H), 7.52 - 7.39 (m, 2H), 7.39 - 7.27 (m, 4H), 4.26 (dd, J= 27.9, 7.3

Hz, 4H), 3.52 (t, J= 9.2 Hz, 1H), 3.05 (d, J= 9.3 Hz, 3H), 2.28 (s, 2H), 1.57 (d, J= 10.8 Hz, 1H), 1.31 (d, J =42.3 Hz, 9H).

Step-2: Préparation of (9H-fluoren-9-yl)methyl (((3S,5S)-5-((6-bromopyridin-2yl)carbamoyl)pyrrolidin-3-yl)methyl)carbamate (212c)

Compound 212c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-ter/-butyl 4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyI)-2-((6bromopyridin-2-yl)carbamoyl)pyrrolidine-l-carboxylate (212b) (220 mg, 0.354 mmol) in DCM (2 mL) using TFA (0.191 mL, 2.478 mmol) and stirring ovemight at RT. This gave after workup TFA sait of (9H-fluoren-9-yI)methyl (((3S,5S)-5-((6-bromopyridin-220 yl)carbamoyI)pyrrolidin-3-yl)methyl)carbamate (212c) (210 mg, 93% yield); ’HNMR (300 MHz, DMSO-îZ₆) δ 11.42 (s, 1H), 9.32 (s, 1H), 8.83 (s, 1H), 8.07 (d, J= 8.2 Hz, 1H), 7.95 7.74 (m, 3H), 7.66 (d, J= 7.4 Hz, 2H), 7.52 - 7.25 (m, 6H), 4.33 (d, J= 6.9 Hz, 3H), 4.20 (t, J= 6.7 Hz, 1H), 3.35 (s, 1H), 3.09 (t, J= 6.0 Hz, 2H), 2.97 (s, 1H), 2.57 - 2.53 (m, 1H), 2.43 (s, 1H), 1.73 - 1.58 (m, 1H); MS (ES+): 521.10 (M+l).

Step-3: Préparation of (9H-fluoren-9-yl)methyl (((3R,5S)-l-(2-(4-amino-6-(trifluoromethyl)9H-pyrimido[4,5-b]indol-9-yl)acetyl)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-3yl)methyl)carbamate (212d)

Compound 212d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (9H-fluoren-9-yl)methyI (((3S,5S)-5-((6-bromopyridin-230 yl)carbamoyl)pyrrolidin-3-yl)methyl)carbamate (212c) (200 mg, 0.315 mmol) in DMF (2 mL) using TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (134 mg, 0.315 mmol), HATU (180 mg, 0.472 mmol), DIPEA (0.274 mL, 1.574 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column

383 chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 50%] (9H-fluoren9-yl)methyl (((3R,5S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-3-yl)methyl)carbamate (212d) (248 mg, 97% yield) as a white solid; ^rH NMR (300 MHz, DMSO-7₆) δ 10.86 (s, 1H), 8.79 (s,

1H), 8.34 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.90 (d, J= 7.3 Hz, 2H), 7.74 - 7.63 (m, 5H),

7.55 (d, J= 4.2 Hz, 3H), 7.43 - 7.29 (m, 5H), 5.41 (d, J= 17.3 Hz, 1H), 5.25 (d, J= 17.1 Hz, 1H), 4.44 (t, J= 8.1 Hz, 1H), 4.37 (d, J= 6.7 Hz, 2H), 4.24 (t, J= 6.8 Hz, 1H), 4.05 (t, J= 8.7 Hz, 1H), 3.50 (t, J= 9.5 Hz, 1H), 3.15 - 3.06 (m, 3H), 2.33 (dt, J= 16.9, 8.7 Hz, 1H), 1.62 (q, 7= 10.1 Hz, 1H).

Step-4: Préparation of (2S,4R)-l-(2-(4-amino-6-(trifhioromethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-4-(aminomethyI)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (212e)

Compound 212e was prepared according to the procedure reported in step-4 of scheme-180, from (9H-fluoren-9-yl)methyl (((3R,5S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-5-((6-bromopyridin-2-yl)carbamoyl)pyrrolidin-3-yl)methyl)carbamate (212d) (240 mg, 0.295 mmol) in DMF (1.2 mL) using piperidine (0.291 mL, 2.95 mmol) and stirring at RT for 1 h. This gave after work up and purification using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-620 (trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-4-(aminomethyl)-N-(6bromopyridin-2-yl)pyrrolidine-2-carboxamide (212e) (65 mg, 37% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-7ô) (a mixture of two rotamers) δ 11.42 and 10.95 (2s, 1H, D₂O exchangeable), 9.07 - 8.93 (m, 3H, 2H D2O exchangeable), 8.71 and 8.68 (2s, 1H), 8.37 (s, 3H, D₂O exchangeable), 7.98 (dd, 7= 8.5, 5.6 Hz, 2H), 7.89 - 7.81 (m, 1H), 7.69 (t, 7 =

8.0 Hz, 1H), 7.42 and 7.31 (2d, 7= 7.7 Hz, 1H), 5.53 (s, 2H), 4.49 (t, 7= 7.8 Hz, 1H), 4.13 (dd, 7= 10.1, 7.0 Hz, 1H), 3.76 (dd, J= 10.2, 7.4 Hz, 1H), 3.19-3.03 (m, 1H), 3.01- 2.86 (m, 1H), 2.81- 2.67 (m, 1H), 2.61 - 2.57 (m, 1H), 1.78 (dt, 7= 12.8, 7.7 Hz, 1H). ¹⁹F NMR (282 MHz, DMSO-7ô) δ -58.60. MS (ES+): 591.2 (M+l); (ES-): 589.1 (M-l); Analysis calculated for C24H22BrF₃N₈O2 2H₂O.1.9HC1: C, 41.38; H, 4.04; Cl, 9.67; N, 16.08; Found: C, 41.53;

H, 3.96; CI, 9.47; N, 15.72.

Scheme 213

Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyrazin-2-yl)-4-fluoro-4-methyIpyrrolidine-2-carboxamide (213d)

Step-1: Préparation of (2S,4R)-/erZ-butyl 2-((6-bromopyrazin-2-yl)carbamoyl)-4-fluoro-45 methylpyrrolidine-1 -carboxylate (213b)

Compound 213b was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4R)-l-(/erZ-butoxycarbonyl)-4-fluoro-4-methylpyrrolidine-2-carboxylic acid (213a) (0.5 g, 2.022 mmol; CAS # 1386458-93-0) in DCM (15 mL) using 1-methyl-lH-imidazole . (0.403 mL, 5.06 mmol), methanesulfonyl chloride (0.188 mL, 2.427 mmol), 6-bromopyrazin10 2-amine (0.352 g, 2.022 mmol; CAS # 54237-53-5) and stirring at RT for 18 h. This gave after workup (2S,4R)-/ert-butyl 2-((6-bromopyrazin-2-yl)carbamoyl)-4-fluoro-4methylpyrrolidine-1-carboxylate (213b) (0.5 g, 61% yield); MS (ES+): 403.10 (M+l).

Step-2: Préparation of (2S,4R)-N-(6-bromopyrazin-2-yl)-4-fluoro-4-methylpyrrolidine-2. carboxamide (213c)

Compound 213c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-ZerZ-butyl 2-((6-bromopyrazin-2-yl)carbamoyl)-4-fluoro-4-methylpyrroIidine1-carboxylate (213b) (0.5 g, 1.240 mmol) in DCM (7 mL) using TFA (0.669 mL, 8.68 mmol) and stirring overnight at RT. This gave after workup and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1%

385 * HCl) from 0-100%] (2S,4R)-N-(6-bromopyrazin-2-yI)-4-fluoro-4-methylpyrrolidine-2carboxamide (213c) (176 mg, 47% yield); Ή NMR (300 MHz, DMSO-î/₆) δ 11.76 (s, 1H), 9.60 (s, 1H), 9.30 (s, 1H), 8.68 (s, 1H), 4.66 (dd, J= 10.8, 7.6 Hz, 1H), 3.66 - 3.51 (m, 1H), 3.41 (dd, J= 35.2, 13.1 Hz, 1H), 2.77 (td, J= 14.6, 7.5 Hz, 1H), 2.27 (ddd, J= 37.3, 14.2, 5 10.9 Hz, 1H), 1.56 (d, J= 21.4 Hz, 3H); MS (ES+): 303.00 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (213d)

Compound 213d was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (2S,4R)-N-(6-bromopyrazin-2-yl)-4-fluoro-4-methylpyrrolidine-210 carboxamide (213c) (48.0 mg, 0.141 mmol) in DMF (1.2 mL) using TFA sait of 2-(4-amino6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (60 mg, 0.141 mmol), HATU (81 mg, 0.212 mmol), DIPEA (0.123 mL, 0.707 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 50%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyrazin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (213d) (35 mg, 42% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 11.67 and 11.33 (2s, 1H, D₂O exchangeable), 9.44 and 9.21 (2s, 1H), 8.99 (s,

1H), 8.73 (s, 2H, D₂O exchangeable), 8.64 (s, 1H), 8.54 (s, 1H), 7.93 - 7.83 (m, 2H), 5.65 (d,

J= 17.4 Hz, 1H), 5.42 (d, J= 17.4 Hz, 1H), 4.65 (dd, J= 10.0, 7.3 Hz, 1H), 4.32 (dd, J= 18.9, 12.0 Hz, 1H), 3.99 (dd,J=35.3, 11.9 Hz, 1H), 2.68 - 2.55 (m, 1H), 2.32-1.99 (m, 1H), 1.64 and 1.54 (2d, J=21.1 Hz, 3H). ¹⁹FNMR(282 MHz, DMSOMô) δ -58.57, -139.00. MS (ES+): 595.1 (M+l); (ES-): 593.0 (M-l); Analysis calculated for C₂3Hi9BrF4NsO₂

2.5H₂O.HC1.: C, 40.81; H, 3.72; Cl, 5.24; N, 16.56; Found: C, 40.91; H, 3.67; Cl, 5.37; N,

16.25.

Scheme 214

386

Préparation of 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6-yl)propanoic acid (214b)

Step-1 : Préparation of ethyl 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5b]indol-6-yl)propanoate (214a)

Compound 214a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(3-ethoxy-3-oxopropyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (45c) (75 mg, 0.164 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (65.6 mg, 0.197 mmol), HATU (94 mg, 0.247 mmol), DIPEA (106 mg, 0.822 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] ethyl 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6yl)propanoate (214a) (78 mg, 76% yield) as a white solid; *H NMR (300 MHz, DMSO-àfo) δ 10.77 (s, 1H), 8.80 - 8.54 (m, 3H), 8.46 - 8.36 (m, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.70 (t, J =

8.0 Hz, 1H), 7.61 (d, J = 8.5 Hz, 1H), 7.45 (dd, J = 8.5, 1.5 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H),

5.70 (d, J = 17.3 Hz, 1H), 5.35 (d, J = 17.2 Hz, 1H), 4.37 (dd, J = 9.0, 6.0 Hz, 1H), 4.05 (q, J = 7.1 Hz, 2H), 3.69 (dd, J = 5.6, 2.4 Hz, 1H), 3.03 (t, J = 7.7 Hz, 2H), 2.75 (t, J = 7.7 Hz, 2H), 2.49 - 2.40 (m, 1H), 2.05 - 1.92 (m, 1H), 1.31 (s, 3H), 1.16 (t, J = 7.1 Hz, 3H), 1.01 (t, J = 5.5 Hz, 1H), 0.93 (dd, J = 5.4, 2.4 Hz, 1H); MS (ES+): 620/622 (M+l), (ES-): 618/620 (M25 1).

387

Step-2: Préparation of 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6-yl)propanoic acid (214b)

Compound 214b was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 3-(4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6-yl)propanoate (214a) (50 mg, 0.081 mmol) in THF (2 mL) and water (1 mL) using 2M aqueous lithium hydroxide hydrate (0.040 mL, 0.081 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [Cl8 column (100 g), eluting with

ACN in water (containing 0.1% HCl) from 0-100%] 3-(4-amino-9-(2-((lR,3S,5R)-3-((6bromopyridin-2-yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9Hpyrimido[4,5-b]indol-6-yl)propanoic acid (214b) (33 mg, 69% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-îZô) (a mixture of two rotamers) δ 10.85 and 10.76 (2s, 1H, D₂O exchangeable), 8.71 (s, 2H, D₂O exchangeable), 8.63 and 8.62 (2s, 1H), 8.41 (s, 1H),

8.00 and 7.94 (2d, J= 8.1 Hz, 1H), 7.69 (td, J= 8.0, 4.5 Hz, 1H), 7.60 (dd, J= 8.6, 2.5 Hz,

1H), 7.45 (d, J= 8.5 Hz, 1H), 7.30 (dd, J= 7.7, 4.9 Hz, 1H), 5.70 (d, J= 17.3 Hz, 1H), 5.35 (d, J = 17.3 Hz, 1H), 4.36 (dd, J= 9.0, 5.9 Hz, 1H), 3.71 - 3.67 (m, 1H), 3.00 (t, J= 7.8 Hz, 2H), 2.67 (t, J= 7.8 Hz, 2H), 2.48-2.39 (m, 1H), 2.05 - 1.92 (m, 1H), 1.31 (s, 3H), 1.01 (t, J = 5.5 Hz, 1H), 0.97 - 0.78 (m, 1H); MS (ES+): 592/594 (M+l), (ES-): 590/592 (M-l);

Analysis calculated for C₂₇H₂₆BrN₇O₄.l.lHC1.2.75H₂O: C, 47.54; H, 4.82; Cl, 5.72; N, 14.37; Found: C, 47.62; H, 4.56; Cl, 5.97; N, 14.24. .

Scheme 215

388

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(dimethylamino)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (215g)

Step-1: Préparation of N-(2-bromo-4-(dimethylamino)phenyl)-2,2,2-trifluoroacetamide (215b)

Compound 215b was prepared according to the procedure reported in step-1 of scheme-46, from 3-bromo-Nl,Nl-dimethylbenzene-l,4-diamine (215a) (1.75 g, 8.14 mmol; CAS #107100-00-5) in DCM (25 mL) using triethylamine (1.40 g, 13.83 mmol) trifluoroacetic acid anhydride (2.56 g, 12.20 mmol) in DCM (5 mL) and stirring at RT for 1 h. This gave after workup N-(2-bromo-4-(dimethylamino)phenyl)-2,2,2-trifluoroacetamide (215b) as a black solid (2.59 g, 102% yield) which was used as such for the next step; 'H NMR (300 MHz, DMSO-Jô) δ 10.92 (s, 1H), 7.17 (d, J = 8.8 Hz, 1H), 6.95 (d, J = 2.8 Hz, 1H), 6.74 (dd, J = 8.9, 2.8 Hz, 1H), 2.93 (s, 6H). ¹⁹F NMR (282 MHz, DMSO-îZ₆) δ -74.03; MS (ES+):

311/313 (M+l), (ES-): 309/311 (M-l).

Step-2: Préparation of 2-amino-5-(dimethylamino)-lH-indole-3-carbonitrile (215c)

389

Compound 215c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-4-(dimethyIamino)phenyl)-2,2,2-trifluoroacetamide (215b) (2.59 g, 8.33 mmol) in DMSO (20 mL) using malononitrile (0.660 g, 9.99 mmol), L-proline (0.192 g, 1.665 mmol), Cul (159 mg, 0.833 mmol) a solution of K2CO3 (2.301 g, 16.65 mmol) in water 5 (20 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [SiCh gel (40 g), eluting with EtOAc in hexane from 0-60%] 2-amino-5(dimethylamino)-lH-indole-3-carbonitrile (215c) (0.94 g, 56% yield) as aplum solid; *H NMR (300 MHz, DMSO-J₆) δ 10.32 (s, 1H), 6.96 (d, J= 8.5 Hz, 1H), 6.56 (s, 2H), 6.48 (d, J = 2.3 Hz, 1H), 6.44 (dd, J= 8.6, 2.4 Hz, 1H), 2.82 (s, 6H); MS (ES+): 201 (M+1), (ES-):

199 (M-1).

Step-3: Préparation ofN6,N6-dimethyl-9H-pyrimido[4,5-b]indole-4,6-diamine (215d)

Compound 215d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5-(dimethylamino)-lH-indole-3-carbonitrile (215c) (0.94 g, 4.69 mmol) in éthanol (30 mL) using formamidine acetate (3.91 g, 37.6 mmol) and heating at 80 °C for 16 75 h. This gave after work up and purification by flash column chromatography [S1O2 gel (24 g), eluting with CMA-80 in DCM from 0-20%] N6,N6-dimethyl-9H-pyrimido[4,5-b]indole-4,6diamine (215d) (0.26 g, 24% yield) as a pale-green solid; 'HNMR (300 MHz, DMSO-t/e) δ 11.42 (s, 1H), 8.17 (s, 1H), 7.56 (d, J = 2.4 Hz, 1H), 7.27 (d, J = 8.8 Hz, 1H), 7.09 (s, 2H), 6.91 (dd, J = 8.8, 2.4 Hz, 1H), 2.93 (s, 6H); MS (ES+): 228 (M+l), (ES-): 226 (M-l).

Step-4: Préparation of tert-butyl 2-(4-amino-6-(dimethylamino)-9H-pyrimido[4,5-b]indol-9yl)acetate (215e)

Compound 215e was prepared according to the procedure reported in step-1 of scheme-1, from N6,N6-dimethyl-9H-pyrimido[4,5-b]indole-4,6-diamine (215d) (0.26 g, 1.144 mmol) in DMF (10 mL) using /er/-butyl 2-bromoacetate (0.268 g, 1.373 mmol), Cs2CO₃ (0.746 g, 25 2.288 mmol) and stirring at RT for 16 h. This gave after work up and purification using flash column chromatography [SiO2 gel (40 g), methanol in DCM from 0-6%] Zer/-butyl 2-(4amino-6-(dimethylamino)-9H-pyrimido[4,5-b]indol-9-yl)acetate (215e) (0.23 g, 59% yield) as a pale-yellow solid; *H NMR (300 MHz, DMSO-î/₆) δ 8.21 (s, 1H), 7.61 (d, J= 2.3 Hz, 1H), 7.36 (d, J= 8.9 Hz, 1H), 7.23 (d, J= 2.8 Hz, 2H), 6.94 (dd, J= 9.0, 2.3 Hz, 1H), 5.02 (s, 30 2H), 2.95 (s, 6H), 1.40 (s, 9H); MS (ES+): 342 (M+1).

Step-5: Préparation of 2-(4-amino-6-(dimethylamino)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (215Q

390 * Compound 215f was prepared according to the procedure reported in step-2 of scheme-1, from iert-butyl 2-(4-amino-6-(dimethylamino)-9H-pyrimido[4,5-b]indol-9-yl)acetate (215e) (230 mg, 0.674 mmol) using TFA (768 mg, 6.74 mmol) in DCM (10 mL) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-6-(dimethyIamino)-9H5 pyrimido[4,5-b]indol-9-yl)acetic acid (215f) (315 mg); *H NMR (300 MHz, DMSO-î/ô) δ 8.54 (s, 3H), 8.02 (s, 1H), 7.71 (d, J = 9.0 Hz, 1H), 7.27 (d, J = 9.8 Hz, 1H), 5.22 (d, J = 4.3 Hz, 2H), 3.09 (s, 6H). ¹⁹F NMR (282 MHz, DMSO-<7₆) δ -74.29; MS (ES+): 286 (M+l), (ES): 284 (M-l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(dimethylamino)-9H-pyrimido[4,510 b]indol-9-yI)acetyI)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (215g)

Compound 215g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(dimethylamino)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (215f) (75 mg, 0.188 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(675 bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (62.5 mg, 0.188 mmol), HATU (86 mg, 0.225 mmol), DIPEA (121 mg, 0.939 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1%

HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(dimethylamino)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (215g) (13 mg, 12% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-J₆/D₂O) δ 8.44 (s, 1H), 8.25 (s, 1H), 7.92 (d, J= 8.2 Hz, 1H), 7.71 - 7.57 (m, 2H), 7.44 (dd, J= 9.0, 2.3 Hz, 1H), 7.28 (d, J= 7.7 Hz, 1H), 5.58 (d, J= 17.5 Hz, 1H), 5.36 (d, J= 17.4 Hz, 1H), 4.37 25 4.22 (m, 1H), 3.67-3.50 (m, 1H), 3.14 (s, 6H), 2.56-2.41 (m, 1H), 1.94 (dd, J= 13.3,6.1

Hz, 1H), 1.27 (s, 3H), 1.00 (t, J= 5.5 Hz, 1H), 0.95 - 0.81 (m, 1H); MS (ES+): 563/565 (M+l), (ES-): 561/563 (M-l).

Scheme 216

391

216g

Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5methyl-2-azabicyclo[3.1.0]hexan-2-yI)-2-oxoethyl)-8-methyl-9H-pyrimido[4,5-b]indole-6carboxylate (216g)

Step-1 : Préparation of methyl 3-bromo-5-methyl-4-(2,2,2-trifluoroacetamido)benzoate (216b)

Compound 216b was prepared according to the procedure reported in step-1 of scheme-46, from methyl 4-amino-3-bromo-5-methylbenzoate (216a) (5.00 g, 20.48 mmol; CAS # 900019-52-5) in DCM (25 mL) using triethylamine (3.52 g, 34.8 mmol) and trifluoroacetic acid anhydride (6.45 g, 30.7 mmol) in DCM (5 mL) and stirring at RT for 1 h. This gave after workup methyl 3-bromo-5-methyl-4-(2,2,2-trifluoroacetamido)benzoate (216b) (7.04 g) as a purple solid; Ή NMR (300 MHz, DMSO-t/e) δ 11.49 (s, 1H), 8.08 (d, J= 1.8 Hz, 1H), 7.96 (d, J = 1.9 Hz, 1H), 3.88 (s, 3H), 2.29 (s, 3H). ^,9F NMR (282 MHz, DMSO-Jd) δ -74.06; MS (ES-): 338/340 (M-l).

Step-2: Préparation of methyl 2-amino-3-cyano-7-methyl-lH-indole-5-carboxylate (216c)

392

Compound 216c was prepared according to the procedure reported in step-1 of scheme-11, from methyl 3-bromo-5-methyl-4-(2,2,2-trifluoroacetamido)benzoate (216b) (7.04 g, 20.70 . mmol) in DMSO (30 mL) using malononitrile (1.641 g, 24.84 mmol), L-proline (0.477 g, 4.14 mmol), Cul (394 mg, 2.070 mmol), a solution of K2CO3 (5.72 g, 41.4 mmol) in water (30 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [S1O2 gel (40 g), eluting with EtOAc in hexane from 0-50%]. methyl 2amino-3-cyano-7-methyl-lH-indole-5-carboxylate (216c) (2.75 g, 58% yield) as an orange solid; Ή NMR (300 MHz, DMSO-J₆) δ 11.18 (s, 1H), 7.60 (d, J = 1.6 Hz, 1H), 7.41 (dd, J = 1.6, 0.9 Hz, 1H), 6.79 (s, 2H), 3.82 (s, 3H), 2.39 (s, 3H). MS (ES+): 230 (M+l), (ES-): 228 (M-l).

Step-3: Préparation of methyl 4-amino-8-methyl-9H-pyrimido[4,5-b]indole-6-carboxyIate (216d)

Compound 216d was prepared according to the procedure reported in step-2 of scheme-29, from methyl 2-amino-3-cyano-7-methyl-lH-indole-5-carboxylate (216c) (2.75 g, 12 mmol) using NH₄OAc (2.77 g, 36.0 mmol), HC(OMe)3 (12.73 g, 120 mmol) and heating at 90 °C for 16 h. This gave after work up methyl 4-amino-8-methyl-9H-pyrimido[4,5-b]indole-6carboxylate (216d) (2.65 g, 86% yield) as apale-yellow solid; 'HNMR (300 MHz, DMSOdè) δ 12.22 (s, 1H), 8.78 (s, 1H), 8.30 (s, 1H), 7.80 (s, 1H), 7.34 (s, 2H), 3.88 (s, 3H), 2.57 (s, 3H); MS (ES+): 257 (M+l), (ES-): 255 (M-l).

Step-4: Préparation of methyl 4-amino-9-(2-(terZ-butoxy)-2-oxoethyl)-8-methyl-9Hpyrimido [4,5 -b] indole-6-carboxylate (216e)

Compound 216e was prepared according to the procedure reported in step-1 of scheme-1, from methyl 4-amino-8-methyl-9H-pyrimido[4,5-b]indole-6-carboxylate (216d) (2.00 g, 7.80 mmol) in DMF (15 mL) using ierZ-butyl 2-bromoacetate (1.827 g, 9.37 mmol), CS2CO3 (5.09 g, 15.61 mmol) and stirring at RT for 16 h. This gave after work up methyl 4-amino-9-(2(terZ-butoxy)-2-oxoethyl)-8-methyl-9H-pyrimido[4,5-b]indole-6-carboxylate (216e) (2.25 g, 78 % yield) as a beige solid; 'H NMR (300 MHz, DMSO-îZ₆) δ 8.82 (d, J= 1.6 Hz, 1H), 8.34 (s, 1H), 7.79 (s, 1H), 7.51 (s, 2H), 5.39 (s, 2H), 3.90 (s, 3H), 2.69 (s, 3H), 1.43 (s, 9H); MS (ES+): 371 (M+l), (ES-): 369 (M-l).

Step-5: Préparation of 2-(4-amino-6-(methoxycarbonyl)-8-methyl-9H-pyrimido[4,5-b]indol9-yl)acetic acid (216f)

393 r Compound 216f was prepared according to the procedure reported in step-2 of scheme-1, from methyl 4-amino-9-(2-(/er/-butoxy)-2-oxoethyl)-8-methyl-9H-pyrimido [4,5 -b] indole-6carboxylate (216e) (1.10 g, 2.97 mmol) in DCM (15 mL) using TFA (3.39 g, 29.7 mmol) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-65 (methoxycarbonyl)-8-methyI-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (216f) (1.65 g); *H NMR (300 MHz, DMSO-ùfe) δ 8.96 (s, 1H), 8.75 - 8.62 (m, 2H), 8.60 (s, 1H), 7.89 (s, 1H), 5.48 (s, 2H), 3.92 (s, 3H), 2.74 (s, 3H); ¹⁹F NMR (282 MHz, DMSO) δ -74.69; MS (ES+): 315 (M+l), (ES-): 313 (M-l).

Step-6: Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-210 yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methyl-9Hpyrimido[4,5-b]indole-6-carboxylate (216g)

Compound 216g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(methoxycarbonyl)-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetic acid (216f) (125 mg, 0.292 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N15 (6-bromopyridin-2-yI)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (97 mg, 0.292 mmol), HATU (133 mg, 0.350 mmol), DIPEA (189 mg, 1.459 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing

0.1% HCl) from 0-100%] methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methyl-9Hpyrimido[4,5-b]indole-6-carboxylate (216g) (105 mg, 61% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-ifc) δ 10.83 (s, 1H, D₂O exchangeable), 8.94 (s, 1H), 8.85 (s, 2H, D₂O exchangeable), 8.66 (s, 1H), 8.01 (d, J = 8.1 Hz, 1H), 7.86 (s, 1H), 7.70 (t, J = 8.0 Hz,

1H), 7.31 (d, J = 7.7 Hz, 1H), 5.91 (d, J = 18.0 Hz, 1H), 5.62 (d, J = 17.9 Hz, 1H), 4.39 (dd, J = 9.0, 6.1 Hz, 1H), 3.90 (s, 3H), 3.71 (dd, J = 5.5, 2.3 Hz, 1H), 2.73 (s, 3H), 2.48 - 2.44 (m, 1H), 1.98 (dd, J =13.3, 6.0 Hz, 1H), 1.31 (s, 3H), 1.03 (t, J = 5.5 Hz, 1H), 0.85 (dd, J = 5.5, 2.4 Hz, 1H); MS (ES+): 592/594 (M+l), (ES-): 590/592 (M-l); Analysis calculated for C₂₇H₂₆BrN₇O₄.0.9HC1.2.5H₂O: C, 48.38; H, 4.80; Cl, 4.76; N, 14.63; Found: C, 48.38; H,

4.71; Cl, 4.94; N, 14.34.

Scheme 217

394

217a

Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methyl-9H-pyrimido[4,5-b]indole-6-carboxylate (217a)

Compound 217a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(methoxycarbonyl)-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetic acid (216f) (125 mg, 0.292 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (93 mg, 0.292 mmol), HATU (133 mg, 0.350 mmol), DIPEA (189 mg, 1.459 mmol) and stirring atRT for 16 h.

This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyI)-8-methyl-9H-pyrimido[4,5-b]indole-615 carboxylate (217a) (110 mg, 65% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-dô) δ 10.83 (s, 1H, D₂O exchangeable), 8.94 (s, 1H), 8.79 - 8.43 (m, 3H, 2H D₂O exchangeable), 8.00 (d, J = 8.2 Hz, 1H), 7.86 (s, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.95 (d, J = 18.0 Hz, 1H), 5.67 (d, J = 18.0 Hz, 1H), 4.42 (dd, J = 9.1, 5.7 Hz, 1H), 3.96-3.89 (mi, 4H), 2.75 (s, 3H), 2.43-2.29 (m, 1H), 2.29-2.11 (m, 1H), 2.01 - 1.87 (m,

1H), 1.18 - 1.04 (m, 1H), 0.77 - 0.63 (m, 1H); MS (ES+): 578/580 (M+l), (ES-): 576/578 (M-l); Analysis calculated for C₂₆H24BrN₇O₄.0.9HC1.2.25H₂O: C, 47.91; H, 4.55; Cl, 4.90; N, 15.04; Found: C, 47.86; H, 4.52; Cl, 4.67; N, 15.08.

Scheme 218 .

395

CO₂Me 217a 218a

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methyl-9H-pyrimido[4,5-b]indole-6-carboxyIic acid (218a)

Compound 218a was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methyl-9H-pyrimido[4,5-b]indole-6-carboxylate (217a) (60 mg, 0.104 mmol) in THF (2 mL) and H₂O (1 mL) using 2 M aqueous LiOH (0.052 mL, 0.104 mmol) and stirring for 16 h at RT. This gave after workup and purification by reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] HCl sait of 4-amino-9-(2-((lR,3S,5R)-3-((6bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methyl-9Hpyrimido[4,5-b]indole-6-carboxylic acid (218a) (15 mg, 26 % yield) as a white solid. ’H NMR (300 MHz, DMSO-i/e) (mixture of two rotamers) δ 12.93 (bs, 1 H, D₂O exchangeable),

10.89 and 10.83 (s, 1H, D₂O exchangeable), 8.91 (s, 1H), 8.60 (d, J = 1.9 Hz, 1H), 8.53 (s,

2H, D₂O exchangeable), 8.02 and 7.96 (d, J = 8.1 Hz, 1H), 7.85 (s, 1H), 7.69 (q, J = 8.0 Hz, 1H), 7.30 (dd, J = 7.7, 5.0 Hz, 1H), 5.94 (dd, J = 17.9, 2.4 Hz, 1H), 5.67 (d, J = 17.9 Hz, 1H), 4.91 and 4.44 (dd, J = 10.1, 4.7 Hz, 1H), 3.97 - 3.91 (m, 1H), 2.79 and 2.74 (s, 3H), 2.44 2.15 (m, 2H), 1.97 - 1.81 (m, 1H), 1.27 - 1.02 (m, 1H), 0.97 and 0.70 (m, 1H); MS (ES+):

564/566 (M+l), (ES-): 562/564 (M-l).

Scheme 219

396

CO₂Me CO₂H ²¹⁶9 219a

Préparation of 4-amino-9-(2-(( 1 R,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methyl-9H-pyrimido[4,5-b]indole-6-carboxylic acid (219a)

Compound 219a was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methyl-9H-pyrimido[4,5-b]indole-6-carboxylate (216g) (64 mg, 0.108 mmol) in THF (2 mL) and water (1 mL) using 2M aqueous lithium hydroxide hydrate (0.054 mL, 0.108 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [C 18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%]4-amino-9-(2-((lR,3S,5R)3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8methyl-9H-pyrimido[4,5-b]indole-6-carboxylic acid (219a) (23 mg, 37 % yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSOX) (mixture of two rotamers) δ 12.97 (bs, 1H, D2O exchangeable), 10.89 and 10.82 (s, 1H, D2O exchangeable), 8.91 (s, 1H), 8.61 (d, J = 2.5 Hz, 1H), 8.60 (s, 2H, D₂O exchangeable), 8.02 and 7.95 (d, J = 8.1 Hz, 1H), 7.85 (s, 1H), 7.69 (q, J = 7.8 Hz, 1H), 7.30 (dd, J = 7.7, 4.9 Hz, 1H), 5.90 (d, J = 18.0 Hz, 1H), 5.62 (d, J = 17.9 Hz, 1H), 4.91 and 4.41 (dd, J = 10.0, 4.7 Hz, 1H), 3.71 - 3.68 (m, 1H), 2.77 and 2.73 (s, 3H), 2.53-2.33 (m, 1H), 2.13 - 1.93 (m, 1H), 1.31 (s, 3H), 1.07-0.98 (m, 1H), 0.91-0.81 (m,

1H); MS (ES+): 578/580 (M+l), (ES-): 576/578 (M-l).

Scheme 220

397

47g 220a

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-8-carboxylic acid (220a)

Compound 220a was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-8-carboxylate (47g) (130 mg, 0.230 mmol) in THF (2 mL) and water (4 mL) using IN aqueous lithium hydroxide hydrate (0.461 mL, 0.461 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-50%] 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-8carboxylic acid (220a) (10 mg, 8% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-cfc) δ 13.28 (s, 1H), 10.70 (s, 1H), 8.61 (d, J= 7.8 Hz, 1H), 8.49 (s, 1H), 7.97 (d, J =

8.2 Hz, 1H), 7.84 (d, J= 7.5 Hz, 1H), 7.67 (t, J= 8.0 Hz, 1H), 7.39 (t, J= 7.8 Hz, 1H), 7.29 (d, J= 7.7 Hz, 1H), 6.00 (d, J= 17.4 Hz, 1H), 5.68 (d, J= 17.3 Hz, 1H), 4.34 (dd, J= 8.8, 5.7 Hz, 1H), 3.87-3.67 (m, 1H), 2.38 - 2.10 (m, 2H), 1.96-1.80 (m, 1H), 1.14-0.97 (m, 1H), 0.89 - 0.77 (m, 1H); MS (ES+): 552.10 (M+l); MS (ES+): 550.10 & 552.10 (M+l), MS (ES): 548.10 & 550.00 (M-l).

Scheme 221

398

(R = Me, Et mixture) 221a 46d 221b

Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylate (221c)

Step-1 : Préparation of methyl 4-amino-9-(2-(tert-butoxy)-2-oxoethyl)-9H-pyrimido[4,5b]indole-5-carboxylate (221a)

Compound 221a was prepared according to the procedure reported in step-1 of scheme-1, from a mixture of methyl/ethyl 4-amino-9H-pyrimido[4,5-b]indole-5-carboxylate (46d) (230 mg) in DMF (15 mL) using ZerZ-butyl 2-bromoacetate (0.168 mL, 1.139 mmol), CS2CO3 (773 mg, 2.374 mmol) and stirring at RT for 14 h. This gave after work up and purification using flash column chromatography [SiO₂gel (25 g), eluting with methanol/ethyl acetate (1:9) in hexanes from 0-50%] methyl 4-amino-9-(2-(tert-butoxy)-2-oxoethyl)-9H-pyrimido[4,5b]indole-5-carboxylate (221a) (76 mg); Ή NMR (300 MHz, DMSO-cfe) δ 8.33 (s, 1H), 7.92 (dd, J= 8.2, 1.1 Hz, 1H), 7.83 (dd, J= 7.8, 1.1 Hz, 1H), 7.56 (t, J= 7.9 Hz, 1H), 7.25 (s, 2H),

5.23 (s, 2H), 4.01 (s, 3H), 1.40 (s, 9H); MS (ES+): 357.20 (M+l).

Step-2: Préparation of 2-(4-amino-5-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (221b)

399

Compound 221b was prepared according to the procedure reported in step-2 of scheme-1, from methyl 4-amino-9-(2-(/er/-butoxy)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5carboxylate (221a) (75 mg, 0.21 mmol) in DCM (10 mL) using TFA (0.486 mL, 6.31 mmol) and stirring at RT for 17 h. This gave after work up 2-(4-amino-5-(methoxycarbonyl)-9H5 pyrimido[4,5-b]indol-9-yl)acetic acid (221b) (99 mg) as TFA sait which was used as such for the next step; MS (ES+): 301.10 (M+l).

Step-3: Préparation of 2methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yI)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5b]indo!e-5-carboxylate (221c)

Compound 221c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-5-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (221b) (63 mg, 0.210 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (84 mg, 0.252 mmol), HATU (160 mg, 0.420 mmol), DIPEA (0.183 mL, 1.05 mmol) and stirring at RT for

16 h. This gave after workup and purification by flash column chromatography [silica gel (12

g), eluting with MeOH/EtOAc (1:9) in hexanes from 0 - 60%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin2-yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,520 b]indole-5-carboxylate (221c) (52 mg, 43% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-riô) δ 10.77 (s, 1H), 8.64 (s, 1H), 8.34 (s, 2H), 8.04 - 7.97 (m, 3H), 7.73 - 7.64 (m, 2H), 7.31 (d, J= 7.7 Hz, 1H), 5.81 (d, J= 17.4 Hz, 1H), 5.46 (d, J= 17.3 Hz, 1H), 4.37 (dd, J= 9.1, 5.9 Hz, 1H), 4.00 (s, 3H), 3.70 (dd, J= 5.5, 2.4 Hz, 1H), 2.54 - 2.39 (m, 1H), 1.98 (dd, J= 13.2, 5.9 Hz, 1H), 1.31 (s, 3H), 1.02 (t, J= 5.5 Hz, 1H), 0.96 (dd, J= 5.3, 2.4

Hz, 1H); MS (ES+): 578.10 & 580.10 (M+l), MS (ES-): 576.10 & 578.10 (M-l); Analysis calculated for C26H24BrN7O4.HCl.2H2O: C, 47.98; H, 4.49; Cl, 5.45; N, 15.06; Found: C, 48.24; H, 4.63; Cl, 5.30; N, 15.14.

Scheme 222

400

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylic acid (222a)

Compound 222a was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylate (221c) (100 mg, containing ethyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyI)-9H-pyrimido[4,5-b]indole-5-carboxylate, ratio: 1:1) in THF (2 mL) and water (4 mL) using IN aqueous lithium hydroxide hydrate (0.346 mL, 0. 346 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-50%] 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,515 b]indole-5-carboxylic acid (222a) (6 mg, 6% yield) HCI sait as a white solid; ’H NMR (300 MHz, DMSO-Jô) δ 10.85 (s, 1H), 8.55 (s, 1H), 8.27 (s, 2H), 8.01 - 7.86 (m, 3H), 7.68 (t, J= 8.0 Hz, 1H), 7.62 (t, J= 7.9 Hz, 1H), 7.30 (d, J= 7.6 Hz, 1H), 5.80 - 5.71 (m, 1H), 5.44 (d, J = 17.0 Hz, 1H), 4.96-4.84 (m, 1 H), 3.77 - 3.67 (m, 1H), 2.46-2.35 (m, 1H), 2.12-1.99 (m, 1H), 1.46 - 1.37 (m, 1H), 1.31 (s, 3H), 0.92 - 0.82 (m, 1H); MS (ES+): 564.10 & 566.10 (M+l), MS (ES-): 562.00 & 564.10 (M-l).

Scheme 223

401

47f

NH₂

223a

Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-8-carboxylate (223a)

Compound 223a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (47f) (152 mg, 0.505 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (168 mg, 0.505 mmol), HATU (288 mg, 0.758 mmol), DIPEA (0.440 mL, 2.53 mmol) and stirring at RT for

19 h. This gave after workup and purification by flash column chromatography [silica gel (12

g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-70%] methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-815 carboxylate (223a) (215 mg, 74% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-ί/ό) δ 10.74 (s, 1H), 8.73 (dd, J= 7.9, 1.2 Hz, 1H), 8.64 (s, 1H), 8.58 (s, 3H), 7.99 (d, J= 8.2 Hz, 1H), 7.87 (dd, J= Ί.Ί, 1.1 Hz, 1H), 7.69 (t, J= 8.0 Hz, 1H), 7.47 (t, J= 7.8 Hz, 1H), 7.30 (d, J= 7.7 Hz, 1H), 5.93 (d, J= 17.5 Hz, 1H), 5.62 (d, J= 17.4 Hz, 1H), 4.31 (dd, J = 9.0, 6.0 Hz, 1H), 3.92 (s, 3H), 3.59 (dd, J= 5.3, 2.2 Hz, 1H), 2.51-2.35 (m, 1H), 1.96 (dd,

J= 13.2, 6.0 Hz, 1H), 1.30 (s, 3H), 1.04 (t, J= 5.5 Hz, 1H), 0.90 (dd, J= 5.5, 2.3 Hz, 1H); MS (ES+): 578.10 & 580.10 (M+l), MS (ES-): 576.10 & 578.10 (M-l); Analysis calculated for C26H24BrN7O₄.0.95HC1.2.25H₂O: C, 47.78; H, 4.54; Cl, 5.15; N, 15.00; Found: C, 47.67; H, 4.37; Cl, 4.86; N, 14.71.

Scheme 224

402

Préparation of 4-amino-9-(2-(( 1 R,3 S,5R)-3 -((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yI)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylic acid (224b)

Step-1 : Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5carboxylate (224a)

Compound 224a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-5-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (221b) (61 mg, containing 2-(4-amino-5-(ethoxycarbonyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid, ratio: 1:1) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (77 mg, 0.242 mmol), HATU (154 mg, 0.404 mmol), DIPEA (0.176 mL, 1.010 mmol) and stirring at RT for 19 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with

MeOH/EtOAc (1:9) in hexanes from 0 - 60%] methyl 4-amino-9-(2-((lR,3S,5R)-3-((6bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5b]indole-5-carboxylate (224a) (115 mg, containing ethyl 4-amino-9-(2-((lR,3S,5R)-3-((6bromopyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5b]indole-5-carboxylate, ratio: 1:1) as a white solid; MS (ES+): 564.10 (M+l).

Step-2: Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylic acid (224b)

Compound 224b was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-225 azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylate (224a) (100 mg, containing ethyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2

403 azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5-carboxylate, ratio:

1:1) in THF (2 mL) and water (4 mL) using IN aqueous lithium hydroxide hydrate (0.354 mL, 0.354 mmol) and stirring at RT for 16 h. This gave after workup and purification by reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-50%] 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-5carboxylic acid (224b) (2 mg, 2% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ 10.85 (s, 1H), 8.56 (s, 1H), 8.28 (s, 2H), 8.00 - 7.88 (m, 3H), 7.74 - 7.57 (m, 2H), 7.31 (d, 1H), 5.80 (d, J= 17.3 Hz, 1H), 5.48 (d, J= 17.2 Hz, 1H), 4.90 (dd, J= 11.5, 3.8

Hz, 1H), 4.03-3.88 (m, 1H), 2.69 - 2.52 (m, 1H), 2.02 - 1.72 (m, 2H), 1.33-1.15 (m, 1H), 1.03 - 0.86 (m, 1H); MS (ES+): 550.10 & 552.10 (M+l), MS (ES-): 548.10 & 550.00 (M-l).

Scheme 225

Br

Préparation of methyl (4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yI)carbamoyl)-275 azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6-yl)carbamate (225e)

Step-1: Préparation of 4-amino-9H-pyrimido[4,5-b]indole-6-carbonyl azide (225a)

To a suspension of 4-amino-9H-pyrimido[4,5-b]indole-6-carboxylic acid (169a) (1 g, 4.38 mmol) in THF (25 mL) and DMF (25.00 mL) was added triethylamine (1.832 pL, 13.15 mmol) and stirred at RT for 15 min. The mixture was treated with diphenyl phosphorazidate

404 (2.93 mL, 13.15 mmol; CAS # 26386-88-9) and stirred at RT for 22 h. The reaction mixture was concentrated to dryness to afford 4-amino-9H-pyrimido[4,5-b]indole-6-carbonyl azide (225a) (4.29 g) which was used as such for the next step; MS (ES+): 254.10 (M+l).

Step-2: Préparation of methyl (4-amino-9H-pyrimido[4,5-b]indol-6-yl)carbamate (225b)

To a suspension of 4-amino-9H-pyrimido[4,5-b]indole-6-carbonyl azide (225a) (111 mg) in toluene (4 mL) was added methanol (6.03 mL, 149 mmol), triethylamine (0.366 mL, 2.63 mmol) and stirred at 115 °C for 4 h in a microwave. The reaction mixture was concentrated in vacuum and purified using flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-10%] to afford methyl (4-amino-9H-pyrimido[4,5-b]indol-610 yl)carbamate (225b) (96 mg) as a white solid; MS (ES+): 258.10 (M+l).

Step-3: Préparation of tert-butyl 2-(4-amino-6-((methoxycarbonyl)amino)-9H-pyrimido[4,5b]indol-9-yl)acetate (225c)

Compound 225c was prepared according to the procedure reported in step-1 of scheme-1, from methyl (4-amino-9H-pyrimido[4,5-b]indol-6-yl)carbamate (225b) (90 mg, 0.350 mmol) in DMF (10 mL) using tert-butyl 2-bromoacetate (0.062 mL, 0.420 mmol) CS2CO3 (285 mg, 0.875 mmol) and stirring at RT for 20 h. This gave after workup tert-butyl 2-(4-amino-6((methoxycarbonyl)amino)-9H-pyrimido[4,5-b]indol-9-yl)acetate (225c) as a brown solid (75 mg) which was used as such for the next step; MS (ES+): 372.20 (M+l).

Step-4: Préparation of 2-(4-amino-6-((methoxycarbonyl)amino)-9H-pyrimido[4,5-b]indol-920 yl)acetic acid (225d)

Compound 225d was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-6-((methoxycarbonyl)amino)-9H-pyrimido[4,5-b]indol-9yl)acetate (225c) (75 mg) in DCM (10 mL) using TFA and stirring at RT. This gave after workup TFA sait of 2-(4-amino-6-((methoxycarbonyl)amino)-9H-pyrimido[4,5-b]indol-925 yl)acetic acid (225d) which was used as such for next step; MS (ES+): 316.10 (M+l).

Step-5: Préparation of methyl (4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6yl)carbamate (225e)

Compound 225e was prepared according to the procedure reported in step-3 of scheme-1, 30 from TFA sait of 2-(4-amino-6-((methoxycarbonyl)amino)-9H-pyrimido[4,5-b]indol-9

405 yl)acetic acid (225d) (64 mg, 0.202 mmol) in DMF (12 mL) using HCI sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (97 mg, 0.303 mmol), HATU (154 mg, 0.404 mmol), DIPEA (0.176 mL, 1.010 mmol) and stirring at RT for 20 h. This gave after workup and purification by flash column chromatography [silica gel (25 g), eluting with MeOH in DCM from 0-5%] followed by purification using reverse phase column chromatography [Cl8 column, eluting with ACN in water (containing 0.1% HCl) from 0-100%] methyl (4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indol-6-yl)carbamate (225e) (52 mg, 44 % yield) HCl sait as a white solid; ‘H NMR (300 MHz, DMSO-J6) δ 10.77 (s,

1H), 9.61 (s, 1H), 8.67 - 8.59 (m, 3H), 8.43 (d, J= 1.9 Hz, 1H), 8.00 (d, J= 8.2 Hz, 1H),

7.70 (t, J= 8.0 Hz, 1H), 7.63 (d, J= 8.8 Hz, 1H), 7.46 (dd, J= 8.8, 1.9 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.74 (d, 17.3 Hz, 1H), 5.40 (d, J= 17.3 Hz, 1H), 4.42 (dd, J= 9.1, 5.5 Hz,

1H), 3.99 - 3.78 (m, 1H), 3.69 (s, 3H), 2.43 - 2.09 (m, 2H), 2.02 - 1.79 (m, 1H), 1.14 - 0.96 (m, 1H), 0.85 - 0.68 (m, 1H); MS (ES+): 579.10 & 581.10 (M+l), MS (ES-): 577.00 &

579.10 (M-l); Analysis calculated for C₂₅H23BrN₈O4.1.15HC1.2.25H2O: C, 45.37; H, 4.36;

N: 16.93; Cl, 6.16. Found: C, 45.49; H, 4.22; N, 16.61; Cl: 5.98.

Scheme 226

406

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(pyridin-4-yl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (226c)

Step-1: Préparation of ethyl 2-(4-amino-6-(pyridin-4-yl)-9H-pyrimido[4,5-b]indol-95 yl)acetate (226a)

Compound 226a was prepared according to the procedure reported in step-1 of scheme-62, from ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (200 mg, 0.573 mmol) in dioxane (18 mL) using pyridin-4-ylboronic acid (106 mg, 0.859 mmol), césium carbonate (280 mg, 0.859 mmol) in water (2.2 mL), Pd(PPh3)2Ch (80 mg, 0.115 mmol) and heating at 100 °C for 13 h under nitrogen. This gave after workup and purification using flash column chromatography [silica gel (25 g), eluting with MeOH in DCM from 0 - 8%] ethyl 2(4-amino-6-(pyridin-4-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (226a) (34 mg, 17% yield) as a white solid; ; Ή NMR (300 MHz, DMSO-îZ₆) δ 8.81 (d, J= 1.7 Hz, 1H), 8.68 - 8.60 (m, 2H), 8.32 (s, 1H), 7.96 - 7.88 (m, 3H), 7.75 (d, J= 8.6 Hz, 1H), 7.54 (s, 2H), 5.29 (s, 2H),

4.15 (q, J= 7.1 Hz, 2H), 1.21 (t, J= 7.1 Hz, 3H); MS (ES+): 348.20 (M+l).

407

Step-2: Préparation of 2-(4-amino-6-(pyridin-4-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (226b)

Compound 226b was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-6-(pyridin-4-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (226a) (32 mg, 5 0.092 mmol) in THF (3 mL) and methanol (3 mL) using a solution of lithium hydroxide hydrate (23.67 mg, 0.553 mmol) in water (3 mL) and stirring at RT for 14 h. This gave after workup 2-(4-amino-6-(pyridin-4-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (226b) which was used as such for the next step; MS (ES+): 320.10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(pyridin-4-yl)-9H-pyrimido[4,5-b]indol10 9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicycIo[3.1.0]hexane-3-carboxamide (226c)

Compound 226c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(pyridin-4-yI)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (226b) (0.029 g, 0.092 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (44 mg, 0.138 mmol), HATU (70 mg, 0.184 mmol), DIPEA (0.080 mL, 0.460 mmol) and stirring at RT for 19 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 10%] followed by purification using reverse phase column chromatography [Cl8 column, eluting with ACN in water (containing 0.1% HCl) from 0-100%] (1R,3S,5R)2-(2-(4-amino-6-(pyridin-4-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-220 yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (226c) (29 mg, 51% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-J₆) δ 10.77 (s, 1H), 9.19 (d, J= 1.8 Hz, 1H), 8.97 (d, J= 6.5 Hz, 2H), 8.70 - 8.53 (m, 5H), 8.25 (dd, J= 8.9, 1.7 Hz, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.89 (d, J= 8.8 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.82 (d, J= 17.4 Hz, 1H), 5.47 (d, J= 17.3 Hz, 1H), 4.42 (dd, J= 9.1, 5.5 Hz, 1H), 4.03 - 3.83 (m, 1H), 2.45 25 2.08 (m, 2H), 2.00 - 1.79 (m, 1H), 1.21 - 0.98 (m, 1H), 0.88 - 0.68 (m, 1H); MS (ES+):

583.10 & 585.10 (M+l), MS (ES-): 581.10 & 583.10 (M-l); Analysis calculated for C28H23BrN₈O₂.2.75HC1.3.0 H₂O: C, 45.58; H, 4.34; N, 15.19; Found: C, 45.81; H, 4.30; N, 15.10.

Scheme 227

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (227c)

Step-1: Préparation of tert-butyl 2-(4-amino-7-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (227a)

Compound 227a was prepared according to the procedure reported in step-1 of scheme-62, from tert-butyl 2-(4-amino-7-bromo-9H-pyrimido[4,5-b]indoI-9-yl)acetate (17c) (400 mg, 1.06 mmol) in dioxane (10 mL) using phenyl boronic acid (194 mg, 1.591 mmol), a solution of 3.3 M aqueous K2CO3 (0.964 mL, 3.18 mmol), Pd(PPh3)2Ch (74 mg, 0.106 mmol) and heating at 100 °C for 16 h under nitrogen. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 4%] tertbutyl 2-(4-amino-7-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (227a) (242 mg, 61% yield) as a pale-brown solid; Ή NMR (300 MHz, DMSO-d₆) δ 8.42 (d, J= 8.2 Hz, 1H), 8.30 (s, 1H), 7.92 (s, 1H), 7.82 (d, 2H), 7.60 (dd, J= 8.1, 1.5 Hz, 1H), 7.51 (t, J= 7.6 Hz, 2H), 7.43 15 7.37 (m, 1H), 7.35 (d, J= 5.5 Hz, 2H), 5.22 (s, 2H), 1.41 (s, 9H); MS (ES+): 375 (M+l).

Step-2: Préparation of 2-(4-amino-7-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (227b)

Compound 227b was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-7-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (227a) (242 mg, 0.646 mmol) in DCM (5 mL) using TFA (737 mg, 6.46 mmol) and stirring at RT for 16 h.

This gave after work up TFA sait of 2-(4-amino-7-phenyl-9H-pyrimido[4,5-b]indol-921436

409 yl)acetic acid (227b) (343 mg) which was used as such for the next step; 'H NMR (300 MHz, DMSO-Je) δ 8.68 - 8.48 (m, 4H), 8.19 (s, 1H), 7.88 (d, J = 1.5 Hz, 1H), 7.87 (s, 1H), 7.79 (dd, J = 8.3, 1.5 Hz, 1H), 7.53 (dd, J = 8.3, 6.9 Hz, 2H), 7.46 - 7.38 (m, 1H), 5.38 (s, 2H). ^I9F NMR (282 MHz, DMSO-cfc) δ -74.50; MS (ES+): 319 (M+l), (ES-): 317 (M-l); Analysis calculated for C29H24BrN7O2.HCL2.5H2O: C, 52.46; H, 4.55; Cl, 5.34; N, 14.77; Found: C, 52.45; H, 4.40; Cl, 5.47; N, 14.76.

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-7-phenyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (227c)

Compound 227c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (227b) (75 mg, 0.173 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (55.3 mg, 0.173 mmol), HATU (79 mg, 0.208 mmol), DIPEA (112 mg, 0.867 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in

DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-phenyl-9H-pyrimido[4,5-b]indoI-9-yI)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (227c) (54 mg, 53% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-t7₆) δ 10.77 (s, 1H, D₂O exchangeable),

8.71 (s, 2H, D₂O exchangeable), 8.64 (s, 1H), 8.60 (d, J = 8.3 Hz, 1H), 8.07 - 7.95 (m, 2H),

7.84 (d, J = 1.5 Hz, 1H), 7.81 (s, 1H), 7.75 (dd, J = 8.2, 1.5 Hz, 1H), 7.66 (t, J = 8.0 Hz, 1H), 7.52 (t, J = 7.6 Hz, 2H), 7.46 - 7.39 (m, 1H), 7.30 (d, J = 7.7 Hz, 1H), 5.85 (d, J = 17.3 Hz, 1H), 5.52 (d, J = 17.2 Hz, 1H), 4.43 (dd, J = 9.1, 5.5 Hz, 1H), 3.99 - 3.90 (m, 1H), 2.40 2.29 (m, 1H), 2.29-2.10 (m, 1H), 1.99-1.81 (m, 1H), 1.16-0.98 (m, 1H), 0.83-0.62 (m,

1H); MS (ES+): 582/584 (M+l), (ES-): 580/582 (M-l).

Scheme 228

410

227b 228a

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-phenyI-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (228a)

Compound 228a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (227b) (75 mg, 0.173 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (57.7 mg, 0.173 mmol), HATU (79 mg, 0.208 mmol), DIPEA (112 mg, 0.867 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with

MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (228a) (68 mg, 66% yield) HCl sait as a white solid; ^lH NMR (300 MHz, DMSO-îZ₆) δ 10.77 (s, 1H, D₂O exchangeable), 8.75 (s, 2H, D₂O exchangeable), 8.64 (s, 1H), 8.60 (d, J = 8.3 Hz, 1H), 7.99 (d, J = 8.1 Hz, 2H), 7.84 (s, 1H), 7.82 - 7.72 (m, 2H), 7.67 (t, J = 8.0 Hz, 1H), 7.53 (t, J = 7.5 Hz, 2H), 7.43 (t, J = 7.3 Hz, 1H), 7.30 (d, J = 7.7 Hz, 1H), 5.81 (d, J = 17.3 Hz, 1H), 5.47 (d, J= 17.3 Hz, 1H), 4.39 (dd, J = 9.1, 6.0 Hz, 1H), 3.74 - 368 (m, 1H), 2.49 - 2.42 (m, 1H), 1.99 (dd, J = 13.2, 5.8 Hz, 1H), 1.32 (s, 3H), 1.03 (t, J = 5.5 Hz, 1H), 0.96 - 0.82 (m, 1H); MS (ES+): 596/598 (M+l); (ES-): 594/596 (M-l); Analysis calculated for

C₃oH₂₆BrN₇0₂.1.15HC1.1.75H₂0: C, 53.78; H, 4.61; Cl, 6.09; N, 14.64; Found: C, 53.70; H, 4.48; Cl, 6.02; N, 14.64.

Scheme 229

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyI-6-phenyl-9H-pyrimido[4,5-b]indoI-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (229c)

Step-1: Préparation of ter/-butyl 2-(4-amino-8-methyl-6-phenyl-9H-pyrimido[4,5-b]indol-95 yl)acetate (229a)

Compound 229a was prepared according to the procedure reported in step-1 of scheme-62, from ZerZ-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (192e) (300 mg, 0.767 mmol) in dioxane (4 mL) using phenyl boronic acid (140 mg, 1.150 mmol), a solution of 3.3 M aqueous K2CO3 (0.697 mL, 2.30 mmol), Pd(PPh₃)2Ch (53.8 mg, 0.077 mmol) and heating at 100 °C for 16 h under nitrogen. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0 3%] terLbutyl 2-(4-amino-8-methyl-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (229a) (200 mg, 67% yield) as a yellow solid; Ή NMR (300 MHz, DMSO-îZô) δ 8.52 - 8.43 (m, 1H), 8.31 (s, 1H), 7.91 - 7.82 (m, 2H), 7.47 (dd, J = 15.8, 8.1 Hz, 5H), 7.34 (t, J = 7.3 Hz, 1H),

5.36 (s, 2H), 2.70 (s, 3H), 1.45 (s, 9H); MS (ES+): 389 (M+l), (ES-): 387 (M-l).

Step-2: Préparation of 2-(4-amino-8-methyl-6-phenyl-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (229b)

412

Compound 229b was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-8-methyl-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (229a) (198 mg, 0.510 mmol) in DCM (5 mL) using TFA (581 mg, 5.10 mmol) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-8-methyl-6-phenyl-9H5 pyrimido[4,5-b]indol-9-yl)acetic acid (229b) (282 mg) which was used as such for the next step; Ή NMR (300 MHz, DMSO-î/ô) δ 8.59 (s, 1H), 8.54 (s, 1H), 8.35 (s, 2H), 7.86 (d, 2H), 7.63 (s, 1H), 7.51 (t, J= 8.4, 6.9 Hz, 2H), 7.38 (t, J= Ί.2 Hz, 1H), 5.46 (s, 2H), 2.76 (s, 3H); ¹⁹F NMR (282 MHz, DMSO) δ -74.28; MS (ES+): 333 (M+l); (ES-): 331 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-phenyl-9H-pyrimido[4,510 b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (229c)

Compound 229c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (229b) (75 mg, 0.168 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(615 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (53.5 mg, 0.168 mmol), HATU (77 mg, 0.202 mmol), DIPEA (109 mg, 0.84 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-phenyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (229c) (82 mg, 82% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/ô) δ 10.84 (s, 1H, D₂O exchangeable), 8.83 (s, 2H, D2O exchangeable), 8.65 (s, 1H), 8.61 (s, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.85 (d, J = 7.6 Hz, 2H), 7.71 (t, J = 8.0 Hz, 1H), 7.65 (s, 1H), 7.51 (t, J = 7.6

Hz, 2H), 7.38 (t, J = 7.4 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.95 (d, J = 17.9 Hz, 1H), 5.67 (d, J = 17.9 Hz, 1H), 4.44 (dd, J = 9.0, 5.7 Hz, 1H), 3.99 - 3.88 (m, 1H), 2.78 (s, 3H), 2.45 - 2.30 (m, 1H), 2.30-2.14 (m, 1H), 2.02- 1.87 (m, 1H), 1.20-0.99 (m, 1H), 0.80 - 0.63 (m, 1H); MS (ES+): 596/598 (M+l); (ES-): 594/596 (M-l); Analysis calculated for

C3oH₂6BrN702.HC1.2.25H₂0: C, 53.50; H, 4.71; Cl, 5.26; N, 14.56; Found: C, 53.29; H, 4.53;

Cl, 5.28; N, 14.47.

Scheme 230

413

229b 230a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-phenyl-9H-pyrimido[4,5-b]indol-9yI)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1,0]hexane-3-carboxamide (230a)

Compound 230a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (229b) (75 mg, 0.168 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yI)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (55.9 mg, 0.168 mmol), HATU (77 mg, 0.202 mmol), DIPEA (109 mg, 0.840 mmol) and stirring at RT for 16

h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-phenyl-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (230a) (68 mg, 66% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/ô) δ 10.84 (s, 1H, D₂O exchangeable), 8.79 (s, 2H, D2O exchangeable), 8.64 (s, 1H), 8.61 (s, 1H), 8.02 (d, J = 8.2 Hz, 1H), 7.85 (d, 2H), 7.71 (t, J = 8.0 Hz, 1H), 7.64 (s, 1H), 7.50 (t, J = 8.3, 6.9 Hz, 2H), 7.42 - 7.34 (m, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.90 (d, J = 18.0 Hz, 1H), 5.62 (d, J = 17.9 Hz, 1H), 4.40 (dd, J = 9.0, 6.1 Hz, 1H), 3.71 (dd, J = 5.5, 2.3 Hz, 1H), 2.76 (s, 3H), 2.5320 2.43 (m, 1H), 1.99 (dd, J = 13.3, 6.1 Hz, 1H), 1.32 (s, 3H), 1.03 (t, J = 5.5 Hz, 1H), 0.86 (dd,

J = 5.4, 2.4 Hz, 1H); MS (ES+): 610/612 (M+l), (ES-): 608/610 (M-l); Analysis calculated for C₃iH28BrN₇O2.1.1HC1.2.25H₂O: C, 53.87; H, 4.90; Cl, 5.64; N, 14.19; Found: C, 53.71; H, 4.64; Cl, 5.59; N, 14.27.

Scheme 231

414

(CF₃CO)₂O

Et₃N

231a

231b

231c

HC(OMe)₃ NH₄OAc

Br'^'CO₂ ^,Bu

Cs₂CO₃

231e

231f

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (231g)

Step-1: Préparation ofN-(2-bromo-6-(trifluoromethyl)pyridin-3-yl)-2,2,2-trifluoroacetamide (231b) ₍

Compound 231b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-6-(trifluoromethyl)pyridin-3-amine (231a) (5 g, 20.75 mmol; CAS # 11751916-1) in DCM (30 mL) using triethylamine (3.57 g, 35.3 mmol), trifluoroacetic acid anhydride (6.54 g, 31.1 mmol) and stirring at RT for 1 h. This gave after workup N-(2bromo-6-(trifluoromethyl)pyridin-3-yI)-2,2,2-trifluoroacetamide (231b) as a plum solid (6.94 g, 99% yield) and was used as such for next step; *H NMR (300 MHz, DMSO-î/ô) δ 11.81 (s, 1H), 8.28 (d, J = 8.1 Hz, 1H), 8.13 (d, J = 8.1 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-îZ₆) δ 66.38, -74.12.

Step-2: Préparation of 2-amino-5-(trifluoromethyl)-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (231c)

Compound 231c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-6-(trifluoromethyl)pyridin-3-yl)-2,2,2-trifluoroacetamide (231b) (6.94 g, 20.59 mmol) in DMSO (30 mL) using malononitrile (1.632 g, 24.71 mmol), L-proline (0.474 g, 4.12 mmol), Cul (0.392 g, 2.059 mmol), a solution of K2CO3 (5.69 g, 41.2 mmol) in water (30 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [S1O2 gel (40 g), eluting with EtOAc in hexane from 0-100%] 2-amino-521436

415 (trifluoromethyl)-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (231c) (3.47 g, 75% yield) as a deep pink solid; ’H NMR (300 MHz, DMSO-<&) δ 11.27 (s, 1H), 7.60 (s, 2H), 7.53 (d, J = 8.1 Hz, 1H), 7.34 (d, J = 8.1 Hz, 1H); ¹⁹F NMR (282 MHz, DMSOX) δ -64.41; MS (ES+): 227 (M+l), (ES-): 225 (M-l).

Step-3: Préparation of 6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4amine (23Id)

Compound 231d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5-(trifluoromethyl)-lH-pÿrrolo[3,2-b]pyridine-3-carbonitrile (231c) (3.47 g, 15.34 mmol) in éthanol (10 mL) using NH4OAC (3.55 g, 46.0 mmol), HC(OMe)₃ (8.14 g, 77 10 mmol) and heating at 90 °C for 16 h. This gave after work up 6-(trifluoromethyl)-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (231d) (1.90 g, 49% yield) as apale-gray solid; ’H NMR (300 MHz, DMSO-cfc) δ 12.49 (s, 1H), 8.43 (s, 1H), 8.03 (d, J = 8.4 Hz, 1H), 7.84 (d, J = 8.4 Hz, 1H). ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -64.04; MS (ES+): 254 (M+l); (ES-): 252 (M-l).

Step-4: Préparation of tert-butyl 2-(4-amino-6-(trifluoromethyl)-9Hpyrido[2',3^,:4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (231e)

Compound 231e was prepared according to the procedure reported in step-1 of scheme-1, from 6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (231d) (1.90 g, 7.50 mmol) in DMF (15 mL) using terZ-butyl 2-bromoacetate (1.610 g, 8.25 mmol),

Cs2CO₃ (4.89 g, 15.01 mmol) and stirring at RT for 16 h. This gave after work up Zert-butyl 2-(4-amino-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (231e) (1.74 g, 63% yield) as a pale gray solid; *H NMR (300 MHz, DMSOX) δ 8.48 (s, 1H), 8.30 (d, J = 8.5 Hz, 1H), 8.19 (s, 1H), 7.94 (d, J = 8.6 Hz, 1H), 6.64 (s, 1H), 5.25 (s, 2H), 1.40 (s, 9H). ¹⁹F NMR (282 MHz, DMSOX) δ -64.07; MS (ES+): 368 (M+l), (ES-):

366 (M-l).

Step-5: Préparation of 2-(4-amino-6-(trifluoromethyl)-9H-pyrido[2^,,3^,:4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetic acid (23If)

Compound 231f was prepared according to the procedure reported in step-2 of scheme-1, from terZ-butyl 2-(4-amino-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin30 9-yl)acetate (231e) (1.00 g, 2.72 mmol) using TFA (3.10 g, 27.2 mmol) in DCM (5 mL) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-6-(trifluoromethyl)21436

416

9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (231f) (1.47 g) as a beige solid; ’H NMR (300 MHz, DMSO-Jd) δ 8.55 (s, 1H), 8.39 (d, J = 8.6 Hz, 1H), 7.98 (d, J = 8.5 Hz, 1H), 5.30 (s, 2H); ^I9F NMR (282 MHz, DMSO-î/₆) δ -64.10, -74.90; MS (ES+): 312 (M+l). Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H5 pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1,0]hexane-3-carboxamide (231g)

Compound 231g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetic acid (231f) (75 mg, 0.176 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (58.7 mg, 0.176 mmol), HATU (80 mg, 0.212 mmol), DIPEA (114 mg, 0.882 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with

ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (231g) (73 mg, 70% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-i/g) δ 10.77 (s, 1H, D₂O exchangeable), 8.74 (bs, 1H, D₂O exchangeable), 8.65 (s, 1H), 8.33 (d, J = 8.6 Hz, 1H), 8.01 (d, J = 8.5 Hz, 2H), 7.70 (t, J = 8.0 Hz, 1H), 7.47 (bs, 1H, D₂O exchangeable), 7.32 (d, J = 7.7

Hz, 1H), 5.79 (d, J = 17.4 Hz, 1H), 5.43 (d, J = 17.3 Hz, 1H), 4.38 (dd, J = 9.1, 5.9 Hz, 1H), 3.62 - 3.52 (m, 1H), 2.49 - 2.42 (m, 1H), 1.99 (dd, J = 13.2, 5.8 Hz, 1H), 1.31 (s, 3H), 1.02 (t, J = 5.5 Hz, 1H), 0.96 - 0.88 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-cfc) δ -64.14; MS (ES+): 589/591 (M+l); (ES-): 587/589 (M-l); Analysis calculated for

C₂₄H₂oBrF₃N₈0₂.0.9HC1.1.75H₂0: C, 44.10; H, 3.76; Cl, 4.88; N, 17.14; Found: C, 44.01; H,

3.64; Cl, 4.96; N, 17.07.

Scheme 232

417

NH₂ nh₂ ^223a 232a

Préparation of 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5-b]indole-8-carboxylic acid (232a)

Compound 232a was prepared according to the procedure reported in step-4 of scheme-17, from methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyI)-9H-pyrimido[4,5-b]indole-8-carboxylate (223a) (140 mg, 0.242 mmol) in THF (2 mL) and water (4 mL) using IN aqueous lithium hydroxide hydrate (0.242 mL, 0.242 mmol) and stirring at RT for 16 h. This gave after workup and purification using reverse phase column chromatography [Cl8 column, eluting with ACN in water (containing 0.1% HCl) from 0-70%] 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-9H-pyrimido[4,5b]indole-8-carboxylic acid (232a) (17 mg, 12% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-Jô) δ 10.71 (s, 1H), 8.69 (d, J= 7.9 Hz, 1H), 8.62 (s, 1H), 8.54 (d, J=

12.5 Hz, 3H), 7.98 (d, J= 8.2 Hz, 1H), 7.89 (d, J= 7.6 Hz, 1H), 7.67 (t, J= 8.1 Hz, 1H), 7.46 (t, J= 7.8 Hz, 1H), 7.29 (d, J= 7.7 Hz, 1H), 5.98 (d, J= 17.4 Hz, 1H), 5.68 (d, J= 17.3 Hz, 1H), 4.31 (dd, J= 9.0, 5.9 Hz, 1H), 3.54 (m, 1H), 2.49-2.32 (m, 1H), 1.95 (dd, J= 13.2, 5.9 Hz, 1H), 1.29 (s, 3H), 1.08 - 0.91 (m, 2H); MS (ES+): 565.10; (ES-): 563.10 (M-l).

Scheme 233

418

Préparation of (lS,3R,5S)-2-(l l-amino-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2c][l,3]oxazine-6-carbonyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (233h)

Step-1 : Préparation of 6-bromo-4-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine (233b)

To a solution of 6-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (233a) (1.02 g, 4.39 mmol; CAS# 784150-41-0) in DMSO (10 mL) at 50 °C was added sodium methanethiolate (1.230 g, 17.55 mmol; CAS # 5188-07-8) in three portions over a 3 h interval and heated at 50 °C overnight. The reaction mixture was poured into water (80 mL) and the solution was allowed to stand for 5 h. The solid obtained was collected by filtration washed with water, dried in vacuo to afford 6-bromo-4-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine (233b) (1.01 g, 4.14 mmol, 94 % yield) as a white solid; ’H NMR (300 MHz, DMSOY₆) δ 12.99 (s, 1H), 8.58 (s, 1H), 6.66 (s, 1H), 2.64 (s, 3H); MS (ES+): 243.95/245.9 (M+l).

Step-2: Préparation of 2-(4-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)phenoI (233c)

419

Compound 233c was prepared according to the procedure reported in step-1 of scheme-62, from 6-bromo-4-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine (233b) (922 mg, 3.78 mmol) in 1,4-dioxane (12 mL) using (2-hydroxyphenyl)boronic acid (625 mg, 4.53 mmol), a solution of K2CO3 (1566 mg, 11.33 mmol) in water (1.5 mL) and PdCh(dppf)-CH2C12 adduct (308 mg, 0.378 mmol) and heating at 80 °C for 6 h under nitrogen. This gave after workup and purification using flash column chromatography [silica gel (40 g), eluting with EtOAc/MeOH (9:1) inhexanes from 0 - 100%] 2-(4-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)phenol (233c) (655 mg, 67% yield) as a yellow solid; ’H NMR (300 MHz, DMSO-î/₆) δ 12.22 (s, 1H), 10.34 (s, 1H), 8.59 (s, 1H), 7.83 (dd, J= 7.9, 1.6 Hz, 1H), 7.28 - 7.14 (m, 1H), 7.02 (dd,

J= 8.6, 1.3 Hz, 2H), 6.92 (t, J= 7.4 Hz, 1H), 2.67 (s, 3H); MS (ES+): 258.0 (M+l).

Step-3: Préparation of ethyl ll-(methylthio)-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2c][l,3]oxazine-6-carboxylate (233d)

A mixture of 2-(4-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)phenol (233c) (470 mg, 1.827 mmol), ethyl 2,2-dibromoacetate (1347 mg, 5.48 mmol) and potassium carbonate (757 mg, 5.48 mmol) in DMF (10 mL) was heated to 100 °C and stirred at 100 °C for 100 min. The reaction mixture was cooled to room température, diluted with EtOAc (200mL), washed with water (lOOmL), brine (lOOmL), dried and concentrated in vacuo. The residue obtained was purified using flash column chromatography [silica gel (12 g), eluting with EtOAc/MeOH (9:1) in hexanes from 0-50%] to provide ethyl 1 l-(methylthio)-6H20 benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2-c][l,3]oxazine-6-carboxylate (233d) (298 mg, 48 % yield) as a white yellow solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 8.68 (s, 1H), 8.01 (dd, J= 7.9, 1.6 Hz, 1H), 7.41 (td, J=7.7, 1.6 Hz, 1H), 7.29 (s, 1H), 7.26 - 7.17 (m, 3H), 4.12-3.98 (m, 2H), 2.71 (s, 3H), 1.01 (t, J= 7.1 Hz, 3H).

Step-4: Préparation of ethyl 1 l-(methylsulfonyl)-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,225 c][l,3]oxazine-6-carboxylate (233e)

To a solution of ethyl ll-(methylthio)-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2c][l,3]oxazine-6-carboxylate (233d) (180 mg, 0.527 mmol) in DCM (35 mL) was added mCPBA (354mg) at 0 °C, warmed to RT and stirred ovemight. An additional amount of mCPBA (98mg) was added, the mixture was stirred at RT for 2 h. The reaction mixture was diluted with DCM (25mL), washed with Sat. NaHCO₃ (40 mL), dried, filtered and concentrated in vacuo to afford ethyl 1 l-(methylsulfonyl)-6H21436

420 benzo[e]pyrimido[5',4':4,5]pynOlo[l,2-c][l,3]oxazine-6-carboxylate (233e) (197 mg, 100 % yield) as a yellow solid which was used for the next step; MS (ES+): 374.10 (M+l);

Step-5: Préparation of 1 l-(methylsulfonyl)-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2c][l,3]oxazine-6-carboxylic acid (233f)

Compound 233f was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 1 l-(methylsulfonyl)-6H-benzo[e]pyrimido[5',4':4,5]pynOlo[l,2-c][l,3]oxazine-6carboxylate (233e) (197 mg, 0.528 mmol) in MeOH (1.5 mL), THF (1.5 mL), acetonitrile (3 mL) using a solution of lithium hydroxide hydrate and stirring at RT. The reaction mixture was concentrated in vacuo and was used for the next step without further purification; MS (ES+): 346.0 (M+l);

Step-6: Préparation of ll-amino-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2-c][l,3]oxazine-6carboxylic acid (233g)

To a solution of 11 -(methylsulfonyl)-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2c][l,3]oxazine-6-carboxylic acid (233f) (182 mg) in 1,4-Dioxane (4 mL) was added ammonium hydroxide (3 mL, 23.11 mmol) and stirred at RT overnight. An additional amount of NH4OH (ImL) was added and the reaction mixture was heated to 50 °C for 5 h. The reaction mixture was concentrated in vacuum and residue obtained was purified by reverse phase column chromatography [Cl8 (50g), eluting with ACN in water (without HCl) from 0100%] to provide 1 l-amino-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2-c][l,3]oxazine-620 carboxylic acid (233g) (90 mg, 61 % yield) as a white solid; MS (ES+): 283.0 (M+l).

Step-7: Préparation of (lS,3R,5S)-2-(l l-amino-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2c][l,3]oxazine-6-carbonyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (233h)

Compound 233h was prepared according to the procedure reported in step-3 of scheme-1, from ll-amino-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2-c][l,3]oxazine-6-carboxylic acid (233g) (40 mg, 0.142 mmol) in DMF (3 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (51 mg, 0.160 mmol), HATU (81 mg, 0.213 mmol), DIPEA (0.123 mL, 0.709 mmol) and stirring at RT for 14 h. This gave after workup and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%](lS,3R,5S)2-(l l-amino-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2-c][l,3]oxazine-6-carbonyl)-N-(621436

421 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (233h) (48 mg, 62.0 % yield) HCI sait as a white solid; *H NMR (300 MHz, DMSO-î/₆) δ 10.71 (d, J= 2.2 Hz, 1H), 8.11 8.06 (m, 1H), 7.98 - 7.85 (m, 1H), 7.69 - 7.51 (m, 2H), 7.34 - 7.24 (m, 2H), 7.24 - 7.09 (m, 4H), 7.00 - 6.92 (m, 1H), 4.48 - 4.26 (m, 1H), 4.24 - 3.99 (m, 1H), 2.34 - 2.03 (m, 2H), 2.03 5 -1.83 (m, 1H), 1.47-1.12 (m, 1H), 0.78 - 0.43 (m, 1H); ’H NMR (300 MHz, DMSOiZ₆/D₂O) δ 8.13 - 8.01 (m, 1H), 7.98 - 7.84 (m, 1H), 7.70 - 7.46 (m, 2H), 7.38 - 7.23 (m, 2H), 7.23 - 7.09 (m, 2H), 7.02 - 6.93 (m, 1H), 4.33 (dd, J= 28.1, 7.0 Hz, 1H), 4.11 (dt, J= 30.4, 7.2 Hz, 1H), 2.33-2.02 (m, 2H), 1.99-1.83 (m, 1H), 1.47-1.12 (m, 1H), 0.78-0.43 (m, 1H). MS (ES+): 546.1, 548.1 (M+l); (ES-): 544.1, 546.1 (M-l); Analysis calculated for:

C₂₅H₂oBrN70₃.0.15HC1.0.5H₂0: C, 53.54; H, 3.80; N, 17.48; Cl, 0.95; Found: C, 53.39; H,

3.70; N, 17.41; Cl, 0.95.

Scheme 234

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrido[2^,,3^,:4,5]pyrrolo[2,315 d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (234a)

Compound 234a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (231f) (75 mg, 0.176 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(620 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (56.2 mg, 0.176 mmol), HATU (80 mg, 0.212 mmol), DIPEA (114 mg, 0.882 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from

0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo [3.1.0]hexane-3 -carboxamide

422 (234a) (67 mg, 66% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î7₆) δ 10.77 (s, 1H, D₂O exchangeable), 8.82 (bs, 1H, D₂O exchangeable), 8.67 (s, 1H), 8.34 (d, J= 8.6 Hz, 1H), 8.06 - 7.97 (m, 2H), 7.70 (t, J= 8.0 Hz, 1H), 7.45 (bs, 1H, D₂O exchangeable), 7.32 (d, J = 7.7 Hz, 1H), 5.84 (d, J= 17.4 Hz, 1H), 5.49 (d, J= 17.3 Hz, 1H), 4.43 (dd, J= 9.1, 5.5 Hz, 5 1H), 3.90 (ddd, J= 7.5, 5.4, 2.3 Hz, 1H), 2.40-2.27 (m, 1H), 2.27 - 2.16 (m, 1H), 1.99-1.86 . (m, 1H), 1.20 - 1.00 (m, 1H), 0.88 - 0.68 (m, 1H); ¹⁹F NMR (282 MHz, DMS0-î/₆) δ -64.12;

MS (ES+): 575/577 (M+l), (ES-): 573/575 (M-l); Analysis calculated for C₂3Hi8BrF₃N₈O₂.1.15HC1.1.75H₂O: C, 42.58; H, 3.52; Cl, 6.28; N, 17.27; Found: C, 42.53; H, 3.36; Cl, 6.33; N, 17.08. .

Préparation of (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoro-l-(2-(2-oxo-3,4-dihydro-lHpyrrolo[3',2':5,6]pyrido[4,3-d]pyrimidin-7(2H)-yl)acetyl)pyrrolidine-2-carboxamide (235a)

To a suspension of ZerZ-butyl (l-(2-((2S,4R)-2-((3-chloro-2-fluorobenzyl)carbamoyl)-415 fluoropyrrolidin-l-yl)-2-oxoethyl)-5-cyano-lH-pyrrolo[2,3-b]pyridin-4-yl)carbamate (144e) (300 mg, 0.524 mmol) in methanol cooled with ice/water was added nickel(II) chloride hexahydrate (31.1 mg, 0.131 mmol), sodium borohydride (119 mg, 3.14 mmol) in several portions over a period of 10 min and stirred at RT for 1 h. The reaction mixture was treated with Nl-(2-aminoethyl)ethane-l,2-diamine (0.113 mL, 1.047 mmol) stirred at RT for 0.5 h and concentrated to dryness. The residue was treated with ethyl acetate (120 mL) and washed with water (60 mL). The aqueous phase was extracted again with ethyl acetate (60 mL). The combined extracts were washed with brine (75 mL), dried, filtered and concentrated in vacuum. The crude product was purified by flash column chromatography [silica gel, eluting with dichloromethane/methanol (1:0 to 19:1)]. The product obtained was dissolved in DCM

423 (10 mL) added TFA (0.077 mL, 1.040 mmol) and at RT for 22 h. The reaction mixture was concentrated to dryness and purified by reverse phase column chromatography using [Cl8 column (26 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] to give (2S,4R)-N-(3-chloro-25 fluorobenzyl)-4-fluoro-1 -(2-(2-oxo-3,4-dihydro-1 H-pyrrolo [3 ',2' :5,6]pyrido[4,3 -d]pyrimidin7(2H)-yl)acetyl)pyrrolidine-2-carboxamide (235a) (58 mg, 22% yield for two steps) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 10.54 (s, 1H, D₂O exchangeable), 9.01 & 8.66 (2t, J-6.0 Hz, 1H, D2O exchangeable), 7.97 (s, 1H), 7.50 7.29 (m, 3H, 1H D₂O exchangeable), 7.16 - 7.03 (m, 1H), 6.95 (d, J= 3.5 Hz, 1H), 6.84 (t, J = 7.9 Hz, 1H), 5.56 - 5.07 (m, 3H), 4.84 - 3.71 (m, 7H), 2.59 - 2.23 (m, 1H), 2.23 - 1.81 (m,

1H); ¹⁹F NMR (282 MHz, DMSO-î/ô) (a mixture of two rotamers) δ -121.34 & -121.74 (1F), -175.94 & -176.13 (1F); MS (ES+): 503.10 & 505.10 (M+l); Analysis calculated for C23H21CIF2N6O3.LO HC1.2.25 H₂O: C, 47.64; H, 4.61; N, 14.49; Cl, 12.23; Found: C, 47.97; H, 4.46; N, 14.55; Cl, 11.93.

Scheme 236

236a

HC(OMe)₃

NH₄OAc, AcOH

236c

236b

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-chIoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (2361)

Step-1: Préparation of 2-amino-5-chloro-lH-indole-3-carbonitrile (236b)

Compound 236b was prepared according to the procedure reported in step-1 of scheme-11, fromN-(4-chloro-2-iodophenyl)-2,2,2-trifluoroacetamide (236a) (6.2 g, 17.74 mmol; CAS #

424

784183-51-3) in DMSO (20 mL) using malononitrile (1.406 g, 21.29 mmol), L-proline (0.409 g, 3.55 mmol), Cul (0.338 g, 1.774 mmol), K2CO3 (4.90 g, 35.5 mmol) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification using flash column chromatography [SiCh gel (40 g), eluting with EtOAc in hexane from 0-50%] 25 amino-5-chloro-lH-indole-3-carbonitrile (236b) (2.02 g, 59% yield) as a yellow solid; 'H

NMR (300 MHz, DMSO-t/₆) δ 10.86 (s, 1H), 7.11 (d, J= 8.3 Hz, 1H), 7.08 (d, .7=2.1 Hz, 1H), 6.97 (s, 2H), 6.90 (dd, J= 8.3, 2.1 Hz, 1H); MS (ES+): 192.0 (M+l); (ES-): 190.0 (M1).

Step-2: Préparation of 6-chloro-9H-pyrimido[4,5-b]indol-4-amine (236c)

Compound 236c was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-5-chloro-lH-indole-3-carbonitrile (236b) (2.0 g, 10.44 mmol) using trimethyl orthoformate (22.84 mL, 209 mmol), AcOH (2.98 mL, 52.2 mmol) and NH4OAC (4.02 g, 52.2 mmol). This gave after workup 6-chloro-9H-pyrimido[4,5-b]indol-4-amine (236c) (2.3 g, 89% yield) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-J₆) δ 11.98 (s, 1H), 8.45 (d,

J= 2.0 Hz, 1H), 8.26 (s, 1H), 7.44 (d, J= 8.6 Hz, 1H), 7.36 (dd, J= 8.6, 2.0 Hz, 1H), 7.30 (s, 2H); MS (ES+): 219.00 (M+l).

Step-3: Préparation of tert-butyl 2-(4-amino-6-chloro-9H-pyrimido[4,5-b]indol-9-yl)acetate (236d)

Compound 236d was prepared according to the procedure reported in step-1 of scheme-1, 20 from 6-chIoro-9H-pyrimido[4,5-b]indol-4-amine (236c) (2.25 g, 9.05 mmol) in DMF (60 mL) using tert-butyl 2-bromoacetate (1.471 mL, 9.95 mmol), CS2CO3 (6.49 g, 19.91 mmol) and stirring at RT for 2 h. This gave after workup and purification using reverse phase column chromatography [C 18 column (150 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] tert-butyl 2-(4-amino-6-chloro-9H-pyrimido[4,5-b]indoI-9-yl)acetate (236d) (1.216 g, 40% yield) as a pale yellow solid; 'HNMR (300 MHz, DMSO-cfc) δ 8.60 (d,

J= 2.0 Hz, 1H), 8.44 (s, 1H), 7.70 (d, J= 8.7 Hz, 1H), 7.51 (dd, J= 8.8, 2.0 Hz, 1H), 5.20 (s, 2H), 1.41 (s, 9H); MS (ES+): 333.10 (M+l).

Step-4: Préparation of 2-(4-amino-6-chloro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (236e)

Compound 236e was prepared according to the procedure reported in step-2 of scheme-1, 30 from tert-butyl 2-(4-amino-6-chloro-9H-pyrimido[4,5-b]indol-9-yl)acetate (236d) (1 g, 3.00 mmol) using 20% TFA in DCM (17.25 mL, 45.1 mmol) and stirring at RT for 16 h. This gave after workup 2-(4-amino-6-chloro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (236e) (1.17 g, 100% yield) TFA sait as apale-yellow solid; 'HNMR (300 MHz, DMSO-î/ô) δ 8.66

425 (d, J= 2.0 Hz, 1H), 8.54 (s, 1H), 8.35 (s, 2H), 7.81 (d, J= 8.8 Hz, 1H), 7.56 (dd, J= 8.8, 2.0

Hz, 1H), 5.25 (s, 2H); MS (ES+): 277.00 (M+l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-chloro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (236f)

Compound 236f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-chloro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (236e) (60mg, 0.154 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (48.9 mg, 0.154 mmol), HATU (88 mg, 0.230 mmol), DIPEA (0.134 mL, 0.768 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCI) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-chloro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (236f) (71 mg, 85% yield) HCl sait as a white solid; ^!H NMR (300 MHz, DMSO-t/ô) δ 10.77 (s, 1H, D2O exchangeable), 8.68 (d, J= 2.0 Hz, 1H), 8.60 (s, 1H), 8.56 (s, 2H, D2O exchangeable), 8.01 (d, J= 8.2 Hz, 1H), 7.77 - 7.67 (m, 2H), 7.58 (dd, J= 8.8, 2.0 Hz, 1H), 7.33 (d, J= 7.7 Hz, 1H), 5.77 (d, J= 17.4 Hz, 1H), 5.42 (d, J= 17.3 Hz, 1H), 4.41 (dd, J= 9.0, 5.5 Hz, 1H), 3.90 (td, J= 6.2, 5.3, 2.3 Hz, 1H), 2.38-2.12 (m, 2H), 1.97-1.85 (m, 1H), 1.07 (dt, J = 9.2, 5.4 Hz, 1H), 0.78 (td, J= 5.2, 2.4 Hz, 1H). MS (ES+): 540.1 (M+l); (ES-): 538.0 (M-l); Analysis calculated for C₂₃Hi9BrClN₇O2 1.75H₂O. O.9HC1.:C, 45.65; H, 3.90; Cl, 11.13; N, 16.20; Found: C, 45.60; H, 3.80; Cl, 11.17; N, 15.84.

Scheme 237

426

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-chloro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yI)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (237a)

Compound 237a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-chloro-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (236e) (60 mg, 0.154 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (51.1 mg, 0.154 mmol), HATU (88 mg, 0.230 mmol), DIPEA (0.134 mL, 0.768 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] ( 1 R,3 S ,5R)-2-(2-(4-amino-6-chIoro-9H-pyrimido [4,5-b] indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (237a) (59 mg, 69% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-Jô) δ 10.77 (s, 1H, D₂O exchangeable), 8.68 (d, J= 2.0 Hz, 1H), 8.67 - 8.54 (m, 3H, 2H D₂O exchangeable), 8.01 (d,

J= 8.2 Hz, 1H), 7.78 - 7.66 (m, 2H), 7.58 (dd, J= 8.7, 1.9 Hz, 1H), 7.33 (d, J= 7.7 Hz, 1H), 5.73 (d, J= 17.4 Hz, 1H), 5.37 (d, J= 17.3 Hz, 1H), 4.37 (dd, J= 9.1, 5.9 Hz, 1H), 3.68 (dd, J=5.5, 2.4 Hz, 1H), 2.57-2.52 (m, 1H), 1.98 (dd, J= 13.3, 5.9 Hz, 1H), 1.31 (s, 3H), 1.01 (t, J= 5.4 Hz, 1H), 0.93 (dd, J= 5.5, 2.4 Hz, 1H). MS (ES+): 554.1(M+1); (ES-): 552.1 (ΜΙ); Analysis calculated for C₂4H₂iBrClN₇O₂ 2H₂O.HC1: C, 45.95; H, 4.18; Cl, 11.30; N,

15.63; Found: C, 46.26; H, 4.12; Cl, 10.92; N, 15.48.

Scheme 238

427

238f

238g

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-chloro-8-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (238g)

Step-1 : Préparation of N-(2-bromo-4-chloro-6-methoxyphenyl)-2,2,2-trifluoroacetamide (238b)

Compound 238b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-4-chloro-6-methoxyaniline (238a) (5 g, 21.14 mmol; CAS # 1261895-84-4) in DCM (75 mL) using triethylamine (5.01 mL, 35.9 mmol), trifluoroacetic acid anhydride (4.41 mL, 31.7 mmol) and stirring at RT for 16 h. This gave after workup N-(2-bromo-410 chloro-6-methoxyphenyl)-2,2,2-trifluoroacetamide (238b) (6.93 g, 99% yield) as a dark solid and was used as such for next step; *H NMR (300 MHz, DMSO-cfe) δ 11.10 (s, 1H), 7.48 (d, J= 2.1 Hz, 1H), 7.32 (d, J= 2.2 Hz, 1H), 3.85 (s, 3H); MS (ES+): 331.90 (M+l).

Step-2: Préparation of 2-amino-5-chloro-7-methoxy-lH-indole-3-carbonitrile (238c)

Compound 238c was prepared according to the procedure reported in step-1 of scheme-11, 15 from N-(2-bromo-4-chloro-6-methoxyphenyl)-2,2,2-trifluoroacetamide (238b) (6.9 g, 20.75

428 mmol) in DMSO (20 mL) using malononitrile (1.568 mL, 24.90 mmol), L-proline (0.478 g, 4.15 mmol), Cul (0.395 g, 2.075 mmol), a solution of K2CO3 (5.74 g, 41.5 mmol) in water (20 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification using flash column chromatography [S1O2 gel (40 g), eluting with EtOAc in hexane from 0-50%] 2-amino-5-chloro-7-methoxy-lH-indole-3-carbonitriIe (238c) (1.85 g, 40% yield) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-70 δ 11.04 (s, 1H), 6.76 (d, J = \.ΊΉζ, 1H), 6.64 (d, J= 1.8 Hz, 1H), 6.55 (s, 2H), 3.88 (s, 3H); MS (ES+): 222.00(M+l). Step-3: Préparation of 6-chloro-8-methoxy-9H-pyrimido[4,5-b]indol-4-amine (238d)

Compound 238d was prepared according to the procedure reported in step-1 of scheme-6, 10 from 2-amino-5-chloro-7-methoxy-lH-indole-3-carbonitrile (238c) (1.8 g, 8.12 mmol) using trimethyl orthoformate (17.77 mL, 162 mmol), AcOH (2.322 mL, 40.6 mmol), NH4OAC (3.13 g, 40.6 mmol) and heating at 100 °C for 1 h. This gave after workup 6-chloro-8methoxy-9H-pyrimido[4,5-b]indol-4-amine (238d) (2.33 g, 93% yield) ACOH sait as a paleyellow solid; MS (ES+): 249.1 (M+l).

Step-4: Préparation of tert-butyl 2-(4-amino-6-chloro-8-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetate (238e)

Compound 238e was prepared according to the procedure reported in step-1 of scheme-1, from AcOH sait of 6-chloro-8-methoxy-9H-pyrimido[4,5-b]indol-4-amine (238d) (1 g, 3.24 mmol) in DMF (25 mL) using tert-butyl 2-bromoacetate (0.503 mL, 3.40 mmol), CS2CO3 (2.64 g, 8.10 mmol) and stirring at RT for 15 h. This gave after workup and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] tert-butyl 2-(4-amino-6-chloro-8-methoxy-9Hpyrimido[4,5-b]indol-9-yl)acetate (238e) (1.175 g, 9% yield) as a pale-yellow solid; 'H NMR (300 MHz, DMSO-7₆) δ 8.48 (s, 1H), 8.23 (d, 7= 1.7 Hz, 1H), 8.17 (s, 2H), 7.17 (d, 7= 1.7

Hz, 1H), 5.23 (s, 2H), 3.94 (s, 3H), 1.42 (s, 9H); MS (ES+): 363.10 (M+l).

Step-5: Préparation of 2-(4-amino-6-chloro-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (238f)

Compound 238f was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-6-chIoro-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (238e) (100 mg, 0.276 mmol) using 20% TFA in DCM (1582 pL, 4.13 mmol) and stirring at RT for

h. This gave after workup 2-(4-amino-6-chloro-8-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetic acid (238f) (115 mg, 99% yield) TFA sait as a pale-yellow solid; *H NMR (300

429

MHz, DMSO-ίήΟ δ 8.48 (s, 1H), 8.23 (d, J= 1.7 Hz, 1H), 8.15 (s, 2H), 7.18 (d, J= 1.7 Hz,

1H), 5.28 (s, 2H), 3.94 (s, 3H); MS (ES+): 307.00 (M+l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-chloro-8-methoxy-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (238g)

Compound 238g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-chloro-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (238f) (55 mg, 0.131 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (41.6 mg, 0.131 mmol), HATU (74.6 mg, 0.196 mmol), DIPEA (0.114 mL, 0.654 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-chloro-8-methoxy-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (238g) (52 mg, 70% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/₆) δ 10.74 (s, 1H, D₂O exchangeable), 8.67 - 8.46 (m, 3H, 2H D₂O exchangeable), 8.26 (d, J= 1.7 Hz, 1H), 8.02 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 7.19 (d, J = 1.7 Hz, 1H), 5.78 (d, J= 17.0 Hz, 1H), 5.54 (d, J= 17.0 Hz, 1H), 4.42 (dd, J= 9.0, 5.6 Hz, 1H), 3.95 (s, 3H), 3.89-3.75 (m, 1H), 2.41-2.15 (m, 2H), 2.00-1.86 (m, 1H), 1.11 (dt, J= 9.3, 5.4 Hz, 1H), 0.74 - 0.56 (m, 1H). MS (ES+): 570.1 (M+l); (ES-): 568.1 (M-l); Analysis calculated for C₂₄H₂iBrClN₇O₃ 2H₂O. 0.9HC1. C, 45.06; H, 4.08; Cl, 10.53; N, 15.33; Found: C, 45.34; H, 3.93; Cl, 10.64; N, 15.21.

Scheme 239

430

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-chloro-8-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (239a)

Compound 239a was prepared according to the procedure reported in step-3 of scheme-1, 5 from TFA sait of 2-(4-amino-6-chloro-8-methoxy-9H-pyrimido[4,5-b]indol-9-yI)acetic acid (238f) (55 mg, 0.131 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (43.5 mg, 0.131 mmol), HATU (74.6 mg, 0.196 mmol), DIPEA (0.114 mL, 0.654 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-chloro-8-methoxy-9Hpyrimido[4,5-b]indol-9-yI)acetyl)-N-(6-bromopyridin-2-yl)-5-methyI-2azabicyclo[3.1.0]hexane-3-carboxamide (239a) (40 mg, 52% yield) HCl sait as a white solid;

Ή NMR (300 MHz, DMSO-î/ô) δ 10.74 (s, 1H, D₂O exchangeable), 8.59 - 8.46 (m, 3H, 2H D₂O exchangeable), 8.25 (d, J= 1.7 Hz, 1H), 8.02 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.32 (d, 7.7 Hz, 1H), 7.18 (d, J= 1.7 Hz, 1H), 5.78 - 5.66 (m, 1H), 5.49 (d, J= 17.0

Hz, 1H), 4.38 (dd, J= 9.0, 6.0 Hz, 1H), 3.94 (s, 3H), 3.63 - 3.61 (m, 1H), 2.47-2.39 (m, 1H), 1.99 (dd, J= 13.3, 5.9 Hz, 1H), 1.31 (s, 3H), 1.05 (t, J= 5.4 Hz, 1H), 0.82 (dd, J= 5.3,

2.4 Hz, 1H). MS (ES+): 584.1 (M+l); (ES-): 582.0 (M-l); Analysis calculated for

C₂₅H₂₃BrClN₇O₃ 1.75H₂O. HCl :C, 45.99; H, 4.25; CI, 10.86; N, 15.02; Found: C, 46.07; H, 4.22; Cl, 10.49; N, 14.90.

Scheme 240

431

240e

240f

240g

Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methoxy-9H-pyrimido[4,5-b]indole-6carboxylate (240g)

Step-1 : Préparation of methyl 3-iodo-5-methoxy-4-(2,2,2-trifluoroacetamido)benzoate (240b)

Compound 240b was prepared according to the procedure reported in step-1 of scheme-46, methyl 4-amino-3-iodo-5-methoxybenzoate (240a) (5 g, 16.28 mmol; CAS # 180624-10-6) in DCM (60 mL) using triethylamine (3.86 mL, 27.7 mmol), trifluoroacetic acid anhydride (3.39 mL, 24.42 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [S1O2 gel (40 g), eluting with EtOAc in hexane from 050%] methyl 3-iodo-5-methoxy-4-(2,2,2-trifluoroacetamido)benzoate (240b) (5.51 g, 84% yield) as a yellow solid; ’H NMR (300 MHz, DMSO-cfe) δ 11.25 (s, 1H), 8.02 (d,J= 1.7 Hz, 1H), 7.61 (d, J= 1.7 Hz, 1H), 3.89 (s, 3H), 3.87 (s, 3H).

Step-2: Préparation of methyl 2-amino-3-cyano-7-methoxy-lH-indole-5-carboxylate (240c)

432

Compound 240c was prepared according to the procedure reported in step-1 of scheme-11, from methyl 3-iodo-5-methoxy-4-(2,2,2-trifluoroacetamido)benzoate (240b) (5.5 g, 13.64 mmol) in DMSO (20 mL) using malononitrile (1.031 mL, 16.37 mmol), L-proline (0.314 g, 2.73 mmol), Cul (0.260 g, 1.364 mmol), a solution of K2CO3 (3.77 g, 27.3 mmol) in water (20 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification using flash column chromatography [S1O2 gel (40 g), EtOAc in hexane from 0-50%] methyl 2-amino-3-cyano-7-methoxy-lH-indole-5-carboxylate (240c) (1.83 g, 55% yield) as a yellow solid; ’H NMR (300 MHz, DMSO-ùfe) δ 11.30 (s, 1H), 7.46 (d, J= 1.3 Hz, 1H), 7.16 (d, J= 1.3 Hz, 1H), 6.66 (s, 2H), 3.93 (s, 3H), 3.84 (s, 3H); MS (ES+): 246.0 (M+l); (ES-): 244.1 (M-l).

Step-3: Préparation of methyl 4-amino-8-methoxy-9H-pyrimido[4,5-b]indole-6-carboxylate (240d)

Compound 240d was prepared according to the procedure reported in step-1 of scheme-6, from methyl 2-amino-3-cyano-7-methoxy-lH-indole-5-carboxylate (240c) (1.8 g, 7.34 mmol) using trimethyl orthoformate (16.06 mL, 147 mmol), AcOH (2.099 mL, 36.7 mmol), NH4OAC (2.83 g, 36.7 mmol) and stirring at 100 °C for 16 h. This gave after workup methyl 4-amino-8-methoxy-9H-pyrimido[4,5-b]indole-6-carboxylate (240d) (1.82 g, 87% yield) AcOH sait as apale-yellow solid; *H NMR (300 MHz, DMSO-t/₆) δ 12.36 (s, 1H), 8.62 (d, J =1.2 Hz, 1H), 8.28 (s, 1H), 7.51 (d, J= 1.3 Hz, 1H), 7.34 (s, 2H), 4.01 (s, 3H), 3.90 (s, 3H);

MS (ES+): 273.10 (M+l).

Step-4: Préparation of methyl 4-amino-9-(2-(ZerZ-butoxy)-2-oxoethyl)-8-methoxy-9Hpyrimido[4,5-b]indole-6-carboxylate (240e)

Compound 240e was prepared according to the procedure reported in step-1 of scheme-1, from AcOH sait of methyl 4-amino-8-methoxy-9H-pyrimido[4,5-b]indole-6-carboxylate (240d) (1 g, 3.67 mmol) in DMF (30 mL) using ZerZ-butyl 2-bromoacetate (0.651 mL, 4.41 mmol), CS2CO3 (2.99 g, 9.18 mmol) and stirring at RT for 30 min. This gave after workup methyl 4-amino-9-(2-(ier/-butoxy)-2-oxoethyl)-8-methoxy-9H-pyrimido[4,5-b]indole-6carboxylate (240e) (1.085 g, 76% yield) as a pale-yellow solid; Ή NMR (300 MHz, DMSO30 d₆) δ 8.65 (d, J= 1.3 Hz, 1H), 8.33 (s, 1H), 7.52 (t, J= 2.0 Hz, 3H), 5.23 (s, 2H), 3.95 (s,

3H), 3.91 (s, 3H), 1.42 (s, 9H); MS (ES+): 387.20 (M+l).

Step-5: Préparation of 2-(4-amino-8-methoxy-6-(methoxycarbonyl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (240f)

433

Compound 240f was prepared according to the procedure reported in step-2 of scheme-1, from methyl 4-amino-9-(2-(ieri-butoxy)-2-oxoethyl)-8-methoxy-9H-pyrimido[4,5-b]indole6-carboxylate (240e) (700 mg, 1.812 mmol) using 20% TFA in DCM (10.40 mL, 27.2 mmol) and stirring at RT for 16 h. This gave after workup 2-(4-amino-8-methoxy-65 (methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (240f) (0.8 g, 99% yield) TFA sait as a pale-yellow solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 8.76 (d, J= 1.3 Hz, 1H), 8.53 (s, 1H), 8.37 (s, 2H), 7.60 (d, J= 1.2 Hz, 1H), 5.35 (s, 2H), 3.98 (s, 3H), 3.93 (s, 3H); MS (ES+): 331.10 (M+l).

Step-6: Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-210 yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methoxy-9H-pyrimido[4,5b]indole-6-carboxylate (240g)

Compound 240g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methoxy-6-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (240£) (75mg, 0.169 mmol) in DMF (1.5 mL) using HCl sait of (1R,3S,5R)-N15 (6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (53.8 mg, 0.169 mmol), HATU (96 mg, 0.253 mmol), DIPEA (0.147 mL, 0.844 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), DMA-80 in DCM from 0 -100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from

0-100%] methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyI)-2azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methoxy-9H-pyrimido[4,5-b]indole-6carboxylate (240g) (70 mg, 70% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-îZô) δ 10.73 (s, 1H, D₂O exchangeable), 8.75 (d, J= 1.2 Hz, 1H), 8.57 (s, 1H), 8.60 8.37 (m, 2H, D₂O exchangeable), 8.02 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.59 (d, J = 1.2 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.82 (d, J= 17.0 Hz, 1H), 5.59 (d, J= 17.0 Hz, 1H),

4.43 (dd, J= 8.9, 5.6 Hz, 1H), 3.99 (s, 3H), 3.92 (s, 3H), 3.89 - 3.82 (m, 1H), 2.39 - 2.15 (m, 2H), 2.01 - 1.85 (m, 1H), 1.18 - 1.03 (m, 1H), 0.73 - 0.61 (m, 1H). MS (ES+): 594.1 (M+l); (ES-): 592.1 (M-l); Analysis calculated for C₂6H₂4BrN?O5 2H₂O. 0.9HC1 :C, 47.08; H, 4.39; Cl, 4.81; N, 14.78; Found: C, 47.15; H, 4.28; Cl, 4.90; N, 14.68.

Scheme 241

Préparation of methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin-2-yl)carbamoyl)-5methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methoxy-9H-pyrimido[4,5-b]indole-6carboxylate (241a)

Compound 241a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methoxy-6-(methoxycarbonyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (240f) (75mg, 0.169 mmol) in DMF (1.5 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (56.1 mg, 0.169 mmol), HATU (96 mg, 0.253 mmol), DIPEA (0.147 mL, 0.844 mmol) and stirring at

RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] methyl 4-amino-9-(2-((lR,3S,5R)-3-((6-bromopyridin2-yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)-8-methoxy-9H- pyrimido[4,5-b]indole-6-carboxylate (241a) (69 mg, 67% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/₆) δ 10.73 (s, 1H, D₂O exchangeable), 8.75 (d, J= 1.3 Hz, 1H), 8.58 (s, 1H), 8.54 (s, 2H, D₂O exchangeable), 8.02 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.58 (d, J= 1.3 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.77 (d, J= 17.0 Hz, 1H), 5.54 (d, J= 16.9 Hz, 1H), 4.39 (dd, J= 8.9, 5.9 Hz, 1H), 3.98 (s, 3H), 3.92 (s, 3H), 3.63 (dd, J= 5.5, 2.3

Hz, 1H), 2.49 - 2.40 (m, 1H), 2.00 (dd, J= 13.2, 5.9 Hz, 1H), 1.32 (s, 3H), 1.12-0.99 (m, 1H), 0.85 - 0.77 (m, 1H); MS (ES+): 608.1 (M+l); (ES-): 606.1 (M-l); Analysis calculated for C₂₇H₂₆BrN₇O₅ 1.5H₂O. 0.9HC1 :C, 48.53; H, 4.51; Cl, 4.77; N, 14.67; Found: C, 48.51; H, 4.47; Cl, 4.66; N, 14.59.

Scheme 242

435

242f 242g

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methoxy-6-nitro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (242g)

Step-1 : Préparation of N-(2-bromo-6-methoxy-4-nitrophenyl)-2,2,2-trifluoroacetamide (242b)

Compound 242b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-6-methoxy-4-nitroaniline (242a) (5 g, 20.24 mmol; CAS # 16618-66-9) in DCM (75 mL) using triethylamine (4.80 mL, 34.4 mmol), trifluoroacetic acid anhydride (4.22 mL, 30.40 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [S1O2 gel (40 g), eluting with EtOAc in hexane from 050%] N-(2-bromo-6-methoxy-4-nitrophenyl)-2,2,2-trifluoroacetamide (242b) (6.7 g, 96% yield) as a yellow solid; MS (ES+): 342.90 (M+l).

Step-2: Préparation of 2-amino-7-methoxy-5-nitro-lH-indole-3-carbonitrile (242c)

Compound 242c was prepared according to the procedure reported in step-1 of scheme-11, 15 from N-(2-bromo-6-methoxy-4-nitrophenyI)-2,2,2-trifluoroacetamide (242b) (6.7 g, 19.53 mmol) in DMSO (20 mL) using malononitrile (1.476 mL, 23.44 mmol), L-proline (0.450 g,

436

3.91 mmol), Cul (0.372 g, 1.953 mmol), K2CO3 (5.40 g, 39.1 mmol) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification using flash column chromatography [S1O2 gel (40 g), eluting with EtOAc in hexane from 0-50%] 2-amino-7methoxy-5-nitro-lH-indole-3-carbonitrile (242c) (290 mg, 6% yield) as a yellow solid; *H

NMR (300 MHz, DMSO-îZ₆) δ 11.71 (s, 1H), 7.69 (d, J= 2.0 Hz, 1H), 7.47 (d, J= 2.0 Hz, 1H), 6.91 (s, 2H), 3.99 (s, 3H).

Step-3: Préparation of 8-methoxy-6-nitro-9H-pyrimido[4,5-b]indol-4-amine (242d)

Compound 242d was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-7-methoxy-5-nitro-lH-indole-3-carbonitrile (242c) (0.28 g, 1.206 mmol) using trimethyl orthoformate (2.64 mL, 24.12 mmol), AcOH (0.345 mL, 6.03 mmol), NH₄OAc (0.465 g, 6.03 mmol) and heating at 100 °C for 16 h. This gave after workup 8-methoxy-6nitro-9H-pyrimido[4,5-b]indol-4-amine (242d) (245 mg, 78% yield) as apale-yellow solid; 'H NMR (300 MHz, DMSO-î/₆) δ 12.73 (s, 1H), 9.05 (d, J= 1.9 Hz, 1H), 8.32 (s, 1H), 7.81 (d, J= 1.9 Hz, 1H), 7.57 (s, 2H), 4.08 (s, 3H); MS (ES+): 260.00 (M+l).

Step-4: Préparation of ZerZ-butyl 2-(4-amino-8-methoxy-6-nitro-9H-pyrimido[4,5-b]indol-9yl)acetate (242e)

Compound 242e was prepared according to the procedure reported in step-1 of scheme-1, from 8-methoxy-6-nitro-9H-pyrimido[4,5-b]indol-4-amine (242d) (0.24 g, 0.926 mmol) in DMF (7 mL) using ZerZ-butyl 2-bromoacetate (0.164 mL, 1.111 mmol), CS2CO3 (0.754 g,

2.315 mmol) and stirring at RT for 30 min. This gave after workup ZerZ-butyl 2-(4-amino-8methoxy-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetate (242e) (155 mg, 45% yield) as a paleyellow solid; Ή NMR (300 MHz, DMSO-î/₆) δ 9.08 (d, J= 1.9 Hz, 1H), 8.38 (s, 1H), 7.83 (d, J= 1.9 Hz, 1H), 7.75 (s, 2H), 5.26 (s, 2H), 4.03 (s, 3H), 1.42 (s, 9H); MS (ES+): 374.17 (M+l).

Step-5: Préparation of 2-(4-amino-8-methoxy-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (242f)

Compound 242f was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-8-methoxy-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetate (242e) (150 mg, 0.402 mmol) using 20% TFA in DCM (2.306 pL, 6.03 mmol) and stirring at RT for 30 16 h. This gave after workup 2-(4-amino-8-methoxy-6-nitro-9H-pyrimido[4,5-b]indol-9yl)acetic acid (242f) (170 mg, 98% yield) TFA sait as a pale-yellow solid; 'H NMR (300

437

MHz, DMSO-îZô) δ 9.15 (d, J= 1.9 Hz, 1H), 8.53 (s, 1H), 8.38 (s, 2H), 7.88 (d, J= 1.9 Hz, 1H), 5.35 (s, 2H), 4.04 (s, 3H); MS (ES+): 318.10 (M+l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methoxy-6-nitro-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide 5 (242g)

Compound 242g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methoxy-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (242f) (60mg, 0.139 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (44.3 mg, 0.139 mmol), 10 HATU (79 mg, 0.209 mmol), DIPEA (0.121 mL, 0.696 mmol) and stirring at RT for 16 h.

This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methoxy-6-nitro-9H-pyrimido[4,5-b]indol15 9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (242g) (31 mg, 38% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-t/ό) δ 10.73 (s, 1H, D₂O exchangeable), 9.15 (d, J= 1.9 Hz, 1H), 8.66 (s, 2H, D₂O exchangeable), 8.61 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.86 (d, J= 1.9 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.82 (d, J= 17.1 Hz, 1H), 5.57 (d, J= 17.0 Hz, 1H), 4.43 (dd, J= 8.9, 5.6 Hz, 1H), 20 4.04 (s, 3H), 3.86 (td, J= 6.2, 5.6, 2.2 Hz, 1H), 2.43-2.13 (m, 2H), 2.01 - 1.83 (m, 1H),

1.19-1.05 (m, 1H), 0.74-0.60 (m, 1H); MS (ES+): 581.1 (M+l); (ES-): 579.1 (M-l);

Analysis calculated for C₂4H₂iBrN₈O₅.1.75H₂O.0.8HCl :C, 44.89; H, 3.97; Cl, 4.42; N, 17.45; Found: C, 44.96; H, 3.81; Cl, 4.49; N, 17.30.

Scheme 243

438

N0₂ ^242f 243a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methoxy-6-nitro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (243a)

Compound 243a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methoxy-6-nitro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (2421) (60mg, 0.139 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (46.3 mg, 0.139 mmol), HATU (79 mg, 0.209 mmol), DIPEA (0.121 mL, 0.696 mmol) and stirring at RT for

J0 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 -100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methoxy-6-nitro-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-315 carboxamide (243a) (49 mg, 59% yield) HCl sait as a pale-yellow solid; *H NMR (300 MHz, DMSOX) δ 10.74 (s, 1H, D₂O exchangeable), 9.15 (d, J= 1.9 Hz, 1H), 8.79 - 8.42 (m, 3H, 2H D₂O exchangeable), 8.02 (d, J= 8.1 Hz, 1H), 7.85 (d, J= 1.9 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.77 (d, J= 17.0 Hz, 1H), 5.52 (d, J= 17.0 Hz, 1H), 4.39 (dd, J = 9.0, 5.9 Hz, 1H), 4.03 (s, 3H), 3.63 (dd, J= 5.6, 2.3 Hz, 1H), 2.48 - 2.41 (m, 1H), 2.00 (dd,

J= 13.3, 5.9 Hz, 1H), 1.32 (s, 3H), 1.11 - 1.00 (m, 1H), 0.90 - 0.77 (m, 1H). MS (ES+): 595.1 (M+l); (ES-): 593.1 (M-l); Analysis calculated for C^H^BrNsOs 1.75H₂O.0.8HCl:C, 45.77; H, 4.19; Cl, 4.32; N, 17.08; Found: C, 45.83; H, 3.98; Cl, 4.65; N, 17.07.

Scheme 244

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (244d)

Step-1: Préparation of ethyl 2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (244a)

Compound 244a was prepared according to the procedure reported in step-1 of scheme-1, from 8-bromo-9H-pyrimido[4,5-b]indol-4-amine (35c) (2 g, 7.60 mmol) in DMF (50 mL) using ethyl 2-bromoacetate (1.012 mL, 9.12 mmol), CS2CO3 (3.72 g, 11.40 mmol) and stirring at RT for 1.5 h. This gave after workup and purification ethyl 2-(4-amino-8-bromo10 9H-pyrimido[4,5-b]indol-9-yI)acetate (244a) (1.35 g, 51% yield) as a pale-yellow solid; ’H

NMR (300 MHz, DMSO-îZ₆) δ 8.40 (dd, J= 7.8, 1.0 Hz, 1H), 8.34 (s, 1H), 7.60 (dd, J= 7.9, 1.0 Hz, 1H), 7.54 - 7.43 (m, 2H), 7.22 (t, J= 7.8 Hz, 1H), 5.55 (s, 2H), 4.18 (q, J= 7.1 Hz, 2H), 1.22 (t, J=1A Hz, 3H); MS (ES+): 349.10 (M+l).

Step-2: Préparation of ethyl 2-(4-amino-8-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (244b)

Compound 244b was prepared according to the procedure reported in step-1 of scheme-62, from ethyl 2-(4-amino-8-bromo-9H-pyrimido[4,5-b]indoI-9-yl)acetate (244a) (500 mg, 1.432 mmol) in dioxane (36 mL) using phenylboronic acid (0.262 g, 2.148 mmol), a solution of césium carbonate (0.700 g, 2.148 mmol) in water (4.5 mL), Pd(PPh3)2Ch (0.201 g, 0.286 mmol) and heating at 100 °C for 16 h under nitrogen. This gave after workup and purification using flash column chromatography [silica gel (40 g), eluting with MeOH/EtOAc (1:9) in

440 hexane from 0-75%] ethyl 2-(4-amino-8-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (244b) (495 mg, 100% yield) as an off-white solid; Ή NMR (300 MHz, DMSO-rf₆) δ 8.41 (dd, J= 7.8, 1.2 Hz, 1H), 8.29 (s, 1H), 7.48 (dd, J= 4.9, 1.8 Hz, 3H), 7.40 (s, 2H), 7.37 - 7.30 (m, 3H), 7.17 (dd, J= 7.4, 1.1 Hz, 1H), 4.65 (s, 2H), 3.89 (q, J= 7.1 Hz, 2H), 1.05 (t, J= 7.1 Hz, 5 3H); MS (ES+): 347.20 (M+l).

Step-3: Préparation of 2-(4-amino-8-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (244c)

Compound 244c was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-8-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (244b) (300 mg, 0.866 mmol) in THF (4.5 mL) and methanol (4.5 mL) using a solution of IN lithium hydroxide hydrate (4.33 mL, 4.33 mmol) and stirring at RT for 19 h. This gave after work up 2-(4amino-8-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (244c) (210 mg, 76% yield) which was used as such for the next step; ’H NMR (300 MHz, DMSO-cfe) δ 12.67 (s, 1H), 8.40 (dd, J= 7.8, 1.2 Hz, 1H), 8.29 (s, 1H), 7.71 - 7.52 (m, 1H), 7.48 (dd, J= 5.0, 2.0 Hz, 3H), 7.43 7.24 (m, 4H), 7.17 (dd, J= 7 A, 1.1 Hz, 1H), 4.53 (s, 2H); MS (ES+): 319.10 (M+l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-phenyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (244d)

Compound 244d was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-8-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (244c) (0.060 g, 0.188 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-220 azabicyclo[3.1.0]hexane-3-carboxamide (4a) (60.1 mg, 0.188 mmol), HATU (108 mg, 0.283 mmol), DIPEA (0.164 mL, 0.942 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), DMA80 in DCM from 0 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(425 amino-8-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (244d) (69 mg, 63% yield) as a white solid; 'H NMR (300 MHz, DMSO-J₆) δ 10.70 (s, 1H, D₂O exchangeable), 8.72 - 8.59 (m, 3H, 2H D₂O exchangeable), 8.55 (dd, J= 7.9, 1.2 Hz, 1H), 8.00 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.60 - 7.37 (m, 5H), 7.37 - 7.19 (m, 3H), 5.28 (d, J= 17.6 Hz, 1H), 4.71 (d, J= 17.6

Hz, 1H), 4.22 (t, J= 7.3 Hz, 1H), 3.18 - 3.07 (m, 1H), 2.13 (dd, J= 8.5, 3.4 Hz, 2H), 1.81 1.63 (m, 1H), 0.68-0.58 (m, 1H), 0.04 --0.08 (m, 1H). MS (ES+): 582.1 (M+l); (ES-): 580.1 (M-l); Analysis calculated for C₂₉H₂₄BrN₇O₂ 1.75H₂O. 0.9HC1 :C, 53.85; H, 4.43; Cl, 4.93; N, 15.16; Found: C, 54.03; H, 4.32; Cl, 5.11; N, 15.11.

244c

441

Scheme 245

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (245a)

Compound 245a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-8-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (244c) (60 mg, 0.188 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyI-2azabicyclo[3.1.0]hexane-3-carboxamide (8a) (62.7 mg, 0.188 mmol), HATU (108 mg, 0.283 mmol), DIPEA (0.164 mL, 0.942 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (245a) (71 mg, 63% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-cfe) δ 10.69 (s, 1H, D₂O exchangeable), 8.63 - 8.41 (m, 4H, 2H D₂O exchangeable), 8.00 (d, J= 8.2 Hz, 1H), 7.68 (t, J= 7.9 Hz, 1H), 7.62 - 7.36 (m, 5H), 7.36 - 7.06 (m, 3H), 5.21 (d, J= 17.9 Hz, 1H), 4.65 (d, J= 17.5 Hz, 1H), 4.17 (dd, J= 8.8, 6.2 Hz, 1H), 2.95 - 2.81 (m, 1H), 2.35 - 2.17 (m, 1H), 1.88 (dd, J= 13.0, 6.2 Hz, 1H), 1.18 (s, 3H), 0.64-0.51 (m, 1H), 0.28-0.12 (m, 1H). MS (ES+): 596.1(M+1); (ES-): 594.1 (M-l); Analysis calculated for C3oH₂6BrN?0₂ 1.75H₂O.HC1 :C, 54.23; H, 4.63; Cl, 5.34; N, 14.76; Found: C, 54.38; H, 4.44; Cl, 5.04; N, 14.68.

Scheme 246

442

Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyI)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-chloropyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (246b)

Compound 246b was prepared according to the procedure reported in step-3 of scheme-1, from HCI sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (50 mg, 0.144 mmol), in DMF (1 mL) using TFA sait of (2S,4R)-N-(6-chloropyridin2-yl)-4-fluoropyrrolidine-2-carboxamide (246a) (51.6 mg, 0.144 mmol; prepared according to the procedure reported by Wiles, Jason A. et al. PCT Int. Appl. (2017), WO 2017035351 Al 20170302.), HATU (82 mg, 0.216 mmol), DIPEA (0.126 mL, 0.721 mmol) and stirring at

RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCI) from 0-100%] (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yI)-4-fluoropynOlidine-215 carboxamide (246b) (51 mg, 66% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-Jé) (a mixture of two rotamers) δ 11.37 and 10.97 (2s, 1H, D2O exchangeable), 9.01 (s, 1H), 8.86 (s, 2H, D2O exchangeable), 8.67 and 8.64 (2s, 1H), 8.22 - 7.71 (m, 4H), 7.30 and 7.19 (2d, J= 7.7 Hz, 1H), 5.76 - 5.01 (m, 3H), 4.62 (dd, J= 9.6, 7.4 Hz, 1H), 4.33 (dd, J = 22.2, 12.5 Hz, 1H), 4.09 (ddd, J= 38.3, 12.5,3.2 Hz, 1H), 2.65-2.54 (m, 1H), 2.31-2.02 (m, 1H). ¹⁹FNMR(282 MHz, DMSO-J₆) δ -58.61, -175.54. MS (ES+): 536.2 (M+l); (ES-):

534.1 (M-l); Analysis calculated for C23H18CIF4N7O22H2O.O.95HCI : C, 45.54; H, 3.81; Cl, 11.40; N, 16.16; Found: C, 45.53; H, 3.72; CI, 11.16; N, 15.83.

Scheme 247

443

247c

Préparation of (2S,4S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yI)-4-methoxypyrrolidine-2-carboxamide (247d)

Step-1: Préparation of (2S,4S)-ieri-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-45 methoxypyrrolidine-l-carboxylate (247b)

Compound 247b was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4S)-l-(tert-butoxycarbonyl)-4-methoxypyrrolidine-2-carboxylic acid (247a) (0.5 g, 2.039 mmol; CAS # 83623-93-2) in DCM (15 mL) using 1-methyl-lH-imidazole (0.406 mL, 5.10 mmol), methanesulfonyl chloride (0.189 mL, 2.446 mmol), 6-bromopyridin-2-amine (0.353 g, 2.039 mmol) and stirring at RT for 18 h. This gave after workup (2S,4S)-terributyl

2-((6-bromopyridin-2-yl)carbamoyl)-4-methoxypyrrolidine-l-carboxylate (247b) (681 mg, 83% yield); Ή NMR (300 MHz, DMSO-î/₆) δ 10.68 (d, J= 27.7 Hz, 1H), 8.09 (q, J= 11.2, 10.0 Hz, 1H), 7.75 (q, J= 7.8 Hz, 1H), 7.35 (d, J= 7.7 Hz, 1H), 4.34 (q, J= 8.5, 7.9 Hz, 1H), 3.96 (t, J= 5.7 Hz, 1H), 3.70 - 3.59 (m, 1H), 3.30 - 3.15 (m, 4H), 2.43 (d, J= 17.1 Hz, 1H),

1.96 - 1.75 (m, 1H), 1.33 (d, J= 42.4 Hz, 9H).

Step-2: Préparation of (2S,4S)-N-(6-bromopyridin-2-yl)-4-methoxypyrrolidine-2carboxamide (247c)

Compound 247c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4S)-tert-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-4-methoxypyrrolidine-1 21436

444 carboxylate (247b) (670 mg, 1.674 mmol) in DCM (9 mL) using TFA (0.903 mL, 11.72 mmol) and stirring overnight at RT. This gave after workup TFA sait of (2S,4S)-N-(6bromopyridin-2-yl)-4-methoxypyrrolidine-2-carboxamide (247c) (0.96 g, 98% yield) as a pale-yellow oil; ’H NMR (300 MHz, DMSO-J₆) δ 11.42 (s, 1H), 9.65 (s, 1H), 8.83 (s, 1H),

8.04 (d, J= 8.3 Hz, 1H), 7.82 (t, J= 8.0 Hz, 1H), 7.45 (dd, J= 7.6, 3.1 Hz, 1H), 4.47 (s, 1H),

4.08 (d, J= 5.2 Hz, 1H), 3.50 - 3.31 (m, 2H), 3.18 (s, 3H), 2.63-2.54 (m, 1H), 2.20 (dd, J= 13.5, 4.7 Hz, 1H); MS (ES+): 300.00 (M+l).

Step-3: Préparation of (2S,4S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-4-methoxypyrrolidine-2-carboxamide (247d)

Compound 247d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (2S,4S)-N-(6-bromopyridin-2-yl)-4-methoxypyrrolidine-2-carboxamide (247c) (84 mg, 0.144 mmol) in DMF (1 mL) using HCl sait of 2-(4-amino-6(trifhioromethyI)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (50 mg, 0.144 mmol), HATU (82 mg, 0.216 mmol), DIPEA (0.151 mL, 0.865 mmol) and stirring at RT for 16 h.

This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4S)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-4-methoxypyrrolidine-2-carboxamide (247d) (55 mg,

64% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-Jé) (a mixture of two rotamers) δ 11.07 and 10.55 (2s, 1H, D₂O exchangeable), 8.98 (s, 1H), 8.74 - 8.47 (m, 3H, 2H D₂O exchangeable), 8.18 - 7.91 (m, 2H), 7.91 - 7.76 (m, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.40 and 7.33 (2d, J= 7.7 Hz, 1H), 5.61 - 5.40 (m, 2H), 4.51 (dd, J= 9.1, 5.1 Hz, 1H), 4.21 4.08 (m, 2H), 3.85 (q, J= 6.4 Hz, 1H), 3.30 and 3.15 (2s, 3H), 2.55-2.45 (m, 1H), 2.12 - 1.96 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-J₆) δ -58.53. MS (ES+): 592.1 (M+l), 614.1 (M+Na);

(ES-): 590.1 (M-l); Analysis calculated for C₂4H₂iBrF₃N₇O3 1.5H₂O.1.15HC1.:C, 43.59; H, 3.83; Cl, 6.17; N, 14.83; Found: C, 43.64; H, 3.61; Cl, 6.14; N, 14.50.

Scheme 248

445

248a ^248b

248c 248d

Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyI)-N-(6-bromopyridin-2-yl)-4-methoxypyrrolidine-2-carboxamide (248d)

Step-1: Préparation of (2S,4R)-terZ-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-45 methoxypyrrolidine-l-carboxylate (248b)

Compound 248b was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4R)-l-(terLbutoxycarbonyl)-4-methoxypyrrolidine-2-carboxylic acid (248a) (0.5 g, 2.039 mmol; CAS # 83624-01-05) in DCM (15 mL) using 1-methyl-IH-imidazole (0.406 mL, 5.10 mmol), methanesulfonyl chloride (0.189 mL, 2.446 mmol), 6-bromopyridin-210 amine (0.353 g, 2.039 mmol) and stirring at RT for 18 h. This gave after workup (2S,4R)terZ-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-4-methoxypyrroIidine-l-carboxylate (248b) (730 mg, 89% yield); Ή NMR (300 MHz, DMSO-î/₆) δ 10.97 (d, J= 18.3 Hz, 1H), 8.10 (dd, J= 14.4, 8.1 Hz, 1H), 7.76 (q, J= 8.2 Hz, 1H), 7.36 (d, J= 7.7 Hz, 1H), 4.39 (q, J= 8.2 Hz, 1H), 3.96 (d, J= 5.1 Hz, 1H), 3.45 (q, J= 3.5 Hz, 2H), 3.23 (s, 3H), 2.39 - 2.23 (m, 1H),

1.93 (ddd, J= 13.4, 8.6, 4.9 Hz, 1H), 1.32 (d, J= 42.4 Hz, 9H); MS (ES+): 400.10 (M+l).

Step-2: Préparation of (2S,4R)-N-(6-bromopyridin-2-yl)-4-methoxypyrrolidine-2carboxamide (248c)

446

Compound 248c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-/erZ-butyl 2-((6-bromopyridin-2-yl)carbamoyl)-4-methoxypyrrolidine-lcarboxylate (248b) (720 mg, 1.799 mmol) in DCM (10 mL) using TFA (0.970 mL, 12.59 mmol) and stirring ovemight at RT. This gave after workup TFA sait of (2S,4R)-N-(65 bromopyridin-2-yl)-4-methoxypyrrolidine-2-carboxamide (248c) (1.05 g, 100% yield) as a pale-yellow oil; ’H NMR (300 MHz, DMSO-î/₆) δ 11.44 (s, 1H), 9.70 (s, 1H), 8.90 (s, 1H), 8.06 (d, J= 8.1 Hz, 1H), 7.82 (t, J= 7.9 Hz, 1H), 7.44 (dd, J= 7.8, 0.7 Hz, 1H), 4.50 - 4.38 (m, 1H), 4.18-4.10 (m, 1H), 3.42-3.32 (m, 2H), 3.27 (s, 3H), 2.65 (dd, J= 13.7, 7.1 Hz, 1H), 2.07 - 1.93 (m, 1H); MS (ES+): 300.0 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-methoxypyrrolidine-2-carboxamide (248d)

Compound 248d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (2S,4R)-N-(6-bromopyridin-2-yl)-4-methoxypyrrolidine-2-carboxamide (248c) (84 mg, 0.144 mmol) in DMF (1 mL) using HCl sait of 2-(4-amino-615 (trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (50 mg, 0.144 mmol), HATU (82 mg, 0.216 mmol), DIPEA (0.151 mL, 0.865 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 -100%] (2S,4R)-l-(2-(4-amino-6-(trifluoromethyl)9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-methoxypyrrolidine-220 carboxamide (248d) (57 mg, 67% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-dô) (a mixture of two rotamers) δ 11.34 and 10.91 (2s, 1H, D₂O exchangeable), 9.00 (s, 1H), 8.77 (s, 2H, D₂O exchangeable), 8.65 and 8.61 (2s, 1H), 8.20 and 7.99 (2d, J= 8.2 Hz, 1H), 7.94 - 7.79 (m, 2H), 7.69 (t, J= 8.0 Hz, 1H), 7.42 and 7.30 (2d, 1H), 5.61 (d, J= 17.3 Hz, 1H), 5.45 (d, J= 17.3 Hz, 1H), 4.52 (t, J= 8.0 Hz, 1H), 4.27 - 4.13 (m, 1H), 4.07 25 3.94 (m,2H), 3.36 and 3.26 (2s,3H), 2.40-2.29 (m, 1H), 2.11 - 1.94 (m, 1H). ^,9FNMR (282 MHz, DMSO-îZô) δ -58.57. MS (ES+): 592.1 (M+l), (ES-): 590.0 (M-l); Analysis calculated for C₂₄H₂iBrF3N7O₃ 1.75H₂O.1.1HC1 :C, 43.41; H, 3.89; Cl, 5.87; N, 14.77; Found: C, 43.38; H, 3.60; Cl, 5.84; N, 14.60.

Scheme 249

447

^Me0 249a

Préparation of ( 1 R,3S,5R)-2-(2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (249a)

Compound 249a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-5-methoxy-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (57f) (60 mg, 0.155 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromo-3methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (10a) (53.8 mg, 0.155 mmol), HATU (89 mg, 0.233 mmol), DIPEA (0.135 mL, 0.777 mmol) and stirring at

RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-5-methoxy-9Hpyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-215 azabicyclo[3.1.0]hexane-3-carboxamide (249a) (52 mg, 59% yield) HCl sait as a white solid;

'H NMR (300 MHz, DMSO-î/ô) δ 10.31 (s, 1H, D₂O exchangeable), 8.57 (s, 1H), 8.56-8.48 (m, 2H, D₂O exchangeable), 8.41 (d, J= 8.7 Hz, 1H), 7.62 (d, J= 8.0 Hz, 1H), 7.45 (d, J = 7.9 Hz, 1H), 7.26 (d, J= 2.3 Hz, 1H), 7.04 (dd, J= 8.7, 2.3 Hz, 1H), 5.63 (d, J= 17.3 Hz, 1H), 5.42 (d, J= 17.3 Hz, 1H), 4.36 (dd, J= 9.3, 5.4 Hz, 1H), 3.78 (s, 3H), 3.69 (dd, J= 5.6,

2.4 Hz, 1H), 2.63-2.53 (m, 1H), 2.09-2.00 (m, 1H), 1.99 (s, 3H), 1.35 (s, 3H), 1.09-1.01 (m, 1H), 1.01-0.90 (m, 1H); MS (ES+): 564.1 (M+l); (ES-): 562.1 (M-l); Analysis calculated for C₂6H₂₆BrN₇O3 1.5H₂O.HC1.:C, 49.73; H, 4.82; CI, 5.65; N, 15.61; Found: C, 49.92; H, 4.81; Cl, 5.80; N, 15.54.

Scheme 250

Préparation of (S)-l-(2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(4cyanobenzyl)pyrrolidine-2-carboxamide (250b)

Compound 250b was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (le) (75 mg, 0.211 mmol) in DMF (5 mL) using HCl sait of (S)-N-(4-cyanobenzyl)pyrrolidine-2-carboxamide (250a) (55.9 mg, 0.211 mmol; CAS # 182291-72-1), HATU (89 mg, 0.233 mmol), DIPEA (136 mg, 1.053 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 6%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (S)-l-(2-(4-amino-9Hpyrimido [4,5-b] indol-9-yl)acetyl)-N-(4-cyanobenzyl)pyrrolidine-2-carboxamide (250b) (55 mg, 58% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-7₆) δ 8.70 - 8.40 (m, 5H, 3H D₂O exchangeable), 7.68 (td, J= 5.2, 4.5, 2.4 Hz, 1H), 7.56 - 7.44 (m, 2H), 7.39 (qd,

J= 7.6, 3.9 Hz, 2H), 7.27 (d, J= 8.1 Hz, 2H), 5.37 (s, 2H), 4.46 - 4.13 (m, 3H), 3.94 - 3.78 (m, 1H), 3.78 - 3.67 (m, 1H), 2.23 - 1.64 (m, 4H); MS (ES+): 454 (M+l); (ES-): 452 (M-l); IR 2233 (cm'¹) Analysis calculated for C₂₅H₂3N₇O₂.1.05HC1.3.5H₂O: C, 54.12; H, 5.64; Cl, 6.71; N, 17.67; Found: C, 54.43; H, 5.27; Cl, 6.90; N, 17.37.

Scheme 251

449

H₂/Pd(OH)₂

251e

Préparation of (S)-3-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(3-chloro-2-fluorobenzyl)-2-oxooxazolidine-4-carboxamide (251f)

Step-1: Préparation of perfluorophenyl 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,55 b]indol-9-yl)acetate (251b)

To a stirred solution of TFA sait of 2-(4-amino-6-(trifhioromethyl)-9H-pyrimido[4,5-b]indoI9-yl)acetic acid (lie) (500 mg, 1.179 mmol) in DMF (2 mL) was added pyridine (0.200 mL, 2.475 mmol) followed by pentafluorophenyl triflouroacetate (251a) (0.253 mL, 1.473 mmol; CAS # 14533-84-7) and stirred for 45 min at RT. The reaction mixture was diluted with ethyl 10 acetate (50 mL), washed with 0.1 N aqueous HCl (2 x 30 mL), 5% aqueous NaHCCh (1 x 30 mL), dried, filtered and concentrated in vacuo to afford perfluorophenyl 2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (251b) (475 mg, 85 % yield) as a white solid and was used as such for the next step; MS (ES+): 477.10 (M+l).

Step-2: Préparation of (S)-benzyl 3-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,515 b]indol-9-yl)acetyl)-2-oxooxazolidine-4-carboxylate (251d)

To a stirred solution of (S)-benzyl 2-oxooxazolidine-4-carboxylate (251c) (218 mg, 0.987 mmol; CAS # 203736-21-4) in dry DMF (1 mL) was added DIPEA (0.345 mL, 1.974 mmol), DMAP (12.06 mg, 0.099 mmol), followed by perfluorophenyl 2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (251b) (470 mg, 0.987 mmol) and 20 stirred for at RT 2 h. The reaction mixture was diluted with ethyl acetate (50 mL), washed with 0.1 N aqueous HCl (2 x 30 mL), 5% aqueous NaHCCh (1 x 30 mL), dried, filtered and

450 concentrated in vacuo. The residue obtained was purified using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 to 50 %] to give (S)-benzyl 3-(2-(4amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-2-oxooxazolidine-4carboxylate (251d) (140mg, 28% yield) as a white solid; Ή NMR (300 MHz, DMSO-î/ô) δ 5 8.84 (s, 1H), 8.34 (s, 1H), 7.69 (d, J= 1.1 Hz, 2H), 7.63 (s, 2H), 7.32 (s, 5H), 5.92 (d, J=

18.6 Hz, 1H), 5.74 (d, J= 18.6 Hz, 1H), 5.17 (s, 2H), 5.06 (dd, J= 9.4, 3.7 Hz, 1H), 4.78 (t, J = 9.3 Hz, 1H), 4.64 (dd, J= 9.2, 3.7 Hz, 1H); MS (ES+): 514.10 (M+l).

Step-3: Préparation of (S)-3-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-2-oxooxazolidine-4-carboxylic acid (251e)

Compound 251e was prepared according to the procedure reported in step-2 of scheme-45, from (S)-benzyl 3-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-2oxooxazolidine-4-carboxylate (251d) (135 mg, 0.263 mmol) in MeOH (30 mL) using palladium hydroxide on carbon (36.9 mg, 0.053 mmol) in a hydrogen atmosphère (balloon) for 2 h at RT. The reaction mixture was filtered and filtrate was concentrated in vacuum to give (S)-3-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-2oxooxazolidine-4-carboxylic acid (251e) (0.070 g, 63% yield) as an off-white solid; ’HNMR (300 MHz, DMSO-dô) δ 8.84 (s, 1H), 8.35 (s, 1H), 7.71 (d, J= 1.1 Hz, 2H), 7.62 (s, 2H), 5.89 (d, J= 18.6 Hz, 1H), 5.74 (d, J= 18.7 Hz, 1H), 4.90-4.80 (m, 1H), 4.75 (t, J= 9.1 Hz, 1H), 4.56 (dd, J= 8.9, 3.3 Hz, 1H); MS (ES+): 424.10 (M+l).

Step-4: Préparation of (S)-3-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-2-oxooxazolidine-4-carboxamide (251f)

Compound 251f was prepared according to the procedure reported in step-3 of scheme-1, from (S)-3-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-2oxooxazolidine-4-carboxylic acid (251e) (30 mg, 0.071 mmol) in DMF (1 mL) using (325 chloro-2-fluorophenyl)methanamine (109b) (8.91 pL, 0.071 mmol; CAS # 182291-72-1), HATU (40.4 mg, 0.106 mmol), DIPEA (0.037 mL, 0.213 mmol) and stirring atRT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl 8 column (30 g), eluting with ACN in water (containing 0.1%

HCl) from 0-100%] (S)-3-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(3-chloro-2-fluprobenzyl)-2-oxooxazolidine-4-carboxamide (251f) (12 mg, 30% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/₆) δ 8.99 - 8.92 (m, 2H), 8.55 (s, 1H), 8.42 (s, 2H, D₂O exchangeable), 7.81 (q, J= 8.7 Hz, 2H), 7.48 - 7.36 (m, 1H), 7.21 21436

451

7.11 (m, 1H), 7.00 (t, J= 7.9 Hz, 1H), 6.01 - 5.73 (m, 2H), 4.86 (dd, J= 9.3, 3.6 Hz, 1H),

4.73 (t, J= 9.1 Hz, 1H), 4.37 (dt, J= 15.1, 4.3 Hz, 3H). ^I9F NMR (282 MHz, DMSO) δ 58.50, -121.33. MS (ES+): 565.1(M+1); (ES-): 563.0 (M-l).

Scheme 252

/0 38d ^252a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1,0]hexane-3-carboxamide (252a)

Compound 252a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (38d) (60 mg, 0.155 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromo-3methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (10a) (53.8 mg, 0.155 mmol), HATU (89 mg, 0.233 mmol), DIPEA (0.135 mL, 0.777 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCI) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-methoxy-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (252a) (50 mg, 57% yield) HCl sait as a white solid; ‘H NMR (300 MHz, DMSO-riô) δ 10.29 (s, 1H, D₂O exchangeable), 8.63 - 8.45 (m, 3H, 2H D₂O exchangeable), 8.06 (d, 2.4 Hz, 1H), 7.62 (d, J= 8.0 Hz, 1H), 7.57 (d, J= 9.0 Hz,

1H), 7.45 (d, J= 7.9 Hz, 1H), 7.15 (dd, J= 8.9, 2.4 Hz, 1H), 5.64 (d, J= 17.3 Hz, 1H), 5.37 (d, J= 17.3 Hz, 1H), 4.36 (dd, J= 9.2, 5.2 Hz, 1H), 3.89 (s, 3H), 3.65 (dd, J= 5.4, 2.4 Hz, 1H), 2.60-2.54 (m, 1H), 2.09-2.03 (m, 1H), 2.01 (s, 3H), 1.33 (s, 3H), 1.08- 1.00 (m, 1H),

452

1.00 - 0.93 (m, 1H). MS (ES+): 564.1 (M+l); (ES-): 562.1 (M-l); Analysis calculated for C26H2₆BrN₇O3 1.75H₂O.HC1.: C, 49.38; H, 4.86; Cl, 5.61; N, 15.50; Found: C, 49.41; H, 4.63; CI, 5.39; N, 15.43.

Scheme 253

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromo-3-methylpyridin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (253a)

Compound 253a was prepared according to the procedure reported in step-3 of scheme-1, 10 from TFA sait of 2-(4-amino-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (36d) (60 mg, 0.155 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromo-3methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (10a) (53.8 mg, 0.155 mmol), HATU (89 mg, 0.233 mmol), DIPEA (0.135 mL, 0.777 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica 15 gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-7-methoxy-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (253a) (46 mg, 53% yield) HCl sait as a white solid; 20 Ή NMR (300 MHz, DMSO-î/₆) δ 10.31 (s, 1H, D₂O exchangeable), 8.67 - 8.46 (m, 3H, 2H

D₂O exchangeable), 8.42 (d, J= 8.7 Hz, 1H), 7.62 (d, J= 8.0 Hz, 1H), 7.45 (d, J= 7.9 Hz, 1H), 7.26 (d, J= 2.3 Hz, 1H), 7.05 (dd, J= 8.7, 2.2 Hz, 1H), 5.63 (d, J= 17.3 Hz, 1H), 5.42 (d, J= 17.2 Hz, 1H), 4.36 (dd, J= 9.3, 5.4 Hz, 1H), 3.78 (s, 3H), 3.69 (dd, J= 5.6, 2.4 Hz, 1H), 2.61-2.53 (m, 1H), 2.04 (dd, J= 13.5, 5.7 Hz, 1H), 1.99 (s, 3H), 1.35 (s, 3H), 1.12

453

0.98 (m, 1H), 0.98 - 0.90 (m, 1H). MS (ES+): 564.2 (M+l); (ES-): 562.2 (M-l); Analysis calculated for C₂6H26BrN₇O3 1.5H₂O.HC1 :C, 49.73; H, 4.82; Cl, 5.65; N, 15.61; Found: C,

49.88; H, 4.80; Cl, 5.77; N, 15.66.

Scheme 254

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (254a)

Compound 254a was prepared according to the procedure reported in step-3 of scheme-1, 10 from TFA sait of 2-(4-amino-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (54f) (60 mg, 0.155 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromo-3methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (10a) (53.8 mg, 0.155 mmol), HATU (89 mg, 0.233 mmol), DIPEA (0.135 mL, 0.777 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica 15 gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methoxy-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (254a) (55 mg, 63% yield) HCl sait as a white solid; 20 ’H NMR (300 MHz, DMSO-cfe) δ 10.28 (s, 1H, D₂O exchangeable), 8.69 - 8.54 (m, 3H, 2H

D₂O exchangeable), 8.10 (d, J= 7.9 Hz, 1H), 7.61 (d, J= 8.0 Hz, 1H), 7.43 (d, J= 7.9 Hz, 1H), 7.36 (t, J= 8.0 Hz, 1H), 7.14 (d, J= 8.1 Hz, 1H), 5.77 (d, J= 17.0 Hz, 1H), 5.56 (d, J = 16.9 Hz, 1H), 4.37 (dd, J= 9.1, 5.3 Hz, 1H), 3.90 (s, 3H), 3.60 (dd, J= 5.6, 2.4 Hz, 1H), 2.60 -2.55 (m, 1H), 2.10-2.03 (m, 1H), 2.01 (s, 3H), 1.34 (s, 3H), 1.14-1.04 (m, 1H), 0.92Ί

454

0.78 (m, 1Η). MS (ES+): 564.1 (M+l); (ES-): 562.1 (M-l); Analysis calculated for C₂₆H₂₆BrN₇O3 1.5H₂O.HC1 : C, 49.73; H, 4.82; Cl, 5.65; N, 15.61; Found: C, 49.71; H, 4.81; Cl, 5.55; N, 15.62.

Scheme 255

TFA

Me

255b

255c

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (255c)

Step-1: Préparation of tert-butyl 2-(4-amino-8-methyl-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-910 yl)acetate (255a)

Compound 255a was prepared according to the procedure reported in step-1 of scheme-59, from terZ-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (192e) (300 mg, 0.767 mmol) in dioxane (4 mL) using/>-tolylboronic acid (156 mg, 1.150 mmol), bis(triphenylphosphine)palladium(II) chloride (53.8 mg, 0.077 mmol) a solution of 3.3 M potassium carbonate (0.697 mL, 2.300 mmol) and stirring at 100 °C for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with

455

MeOH in DCM from 0-3%] ZerZ-butyl 2-(4-amino-8-methyl-6-(p-tolyl)-9H-pyrimido[4,5b]indol-9-yl)acetate (255a) (190 mg, 62% yield) as a white solid; *H NMR (300 MHz, DMSO-îZ₆) δ 8.44 (d, 1.7 Hz, 1H), 8.30 (s, 1H), 7.78 (s, 1H), 7.76 (s, 1H), 7.48 (d, J= 1.7

Hz, 1H), 7.42 (s, 2H), 7.30 (s, 1H), 7.27 (s, 1H), 5.35 (s, 2H), 2.69 (s, 3H), 2.36 (s, 3H), 1.44 5 (s, 9H); MS (ES+): 403 (M+l), (ES-): 401 (M-l).

Step-2: Préparation of 2-(4-amino-8-methyl-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (255b)

Compound 255b was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-8-methyl-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (255a) (190 mg, 0.472 mmol) in DCM (5 mL) using TFA (538 mg, 4.72 mmol) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-8-methyl-6-(p-tolyl)-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (255b) (250 mg); Ή NMR (300 MHz, DMSO-ô/₆) δ 8.77 - 8.51 (m, 4H), 7.77 (d, 2H), 7.64 (s, 1H), 7.32 (d, J= 8.0 Hz, 2H), 5.47 (s, 2H), 2.75 (s, 3H), 2.38 (s, 3H); ^I9F NMR (282 MHz, DMSO) δ -74.46; MS (ES+): 347 (M+l), (ES-): 345 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(p-tolyl)-9H-pyrimido[4,5b]indol-9-yI)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (255c)

Compound 255c was prepared according to the procedure reported in step-3 of scheme-1, 20 from TFA sait of 2-(4-amino-8-methyl-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (255b) (75 mg, 0.163 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (54.2 mg, 0.163 mmol), HATU (74.3 mg, 0.195 mmol) DIPEA (105 mg, 0.814 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12

g), MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9yI)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (255c) (65 mg, 64% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-dô) δ 10.84 (s, 1H, D₂O exchangeable), 8.72 (bs, 2H, D₂O exchangeable), 8.62 (s, 1H), 8.57 (s, 1H), 8.02 (d, J = 8.2 Hz, 1H), 7.81 - 7.65 (m, 3H), 7.61 (s, 1H), 7.38 - 7.21 (m, 3H), 5.87 (d, J = 17.9 Hz, 1H), 5.61 (d, J = 17.9 Hz, 1H), 4.40 (dd, J = 9.0, 6.2 Hz, 1H), 3.74 - 3.67 (m, 1H), 2.74 (s, 3H), 2.49 - 2.43 (m, 1H), 2.37 (s, 3H), 1.99 (dd, J = 13.3, 6.1 Hz, 1H), 1.31 (s, 3H), 1.03

456 (t, J = 5.5 Hz, 1H), 0.85 (dd, J = 5.6, 2.4 Hz, 1H); MS (ES+) 624/626 (M+l), (ES-) 622/624 (M-l); Analysis calculated for C3₂H₃oBrN₇0₂.l.lHC1.2.25H₂0: C, 54.50; H, 5.09; Cl, 5.53; N, 13.90; Found: C, 54.21; H, 5.00; Cl, 5.39; N, 13.87.

Scheme 256

256a ₅ 255b

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (256a)

Compound 256a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (255b) (75 mg, 0.163 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (51.9 mg, 0.163 mmol), HATU (74.3 mg, 0.195 mmol), DIPEA (105 mg, 0.814 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(p-tolyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1,0]hexane-3-carboxamide (256a) (70 mg, 70% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ê) δ 10.83 (s, 1H, D₂O exchangeable), 8.80 (s, 2H, D₂O exchangeable), 8.63 (s, 1H), 8.57 (s, 1H), 8.01 (d, J=

8.1 Hz, 1H), 7.81 - 7.66 (m, 3H), 7.61 (s, 1H), 7.37 - 7.21 (m, 3H), 5.93 (d, J= 18.0 Hz,

1H), 5.64 (d, 17.9 Hz, 1H), 4.43 (dd, J= 9.1, 5.7 Hz, 1H), 3.98 - 3.87 (m, 1H), 2.75 (s,

3H), 2.42 - 2.29 (m, 4H), 2.29-2.12 (m, 1H), 2.00-1.81 (m, 1H), 1.17-0.99 (m, 1H), 0.79 - 0.60 (m, 1H); MS (ES+): 610/612 (M+l), (ES-): 608/610 (M-l); Analysis calculated for

457

C3iH28BrN₇O₂.1.15 HC1.1.75 H₂0: C, 54.44; H, 4.81; Cl, 5.96; N, 14.34; Found: C, 54.27; H,

4.91; Cl, 5.93; N, 14.46.

Scheme 257

257a (CF₃CO)₂O

Et₃N

OMe

NH₄OAc

HC(OMe)₃

257b

257c

OMe

257d 257e

HATU, DIPEA

257f 257g

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methoxy-8-methyI-9H-pyrimido[4,5-b]indoI-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (257g)

Step-1 : Préparation of N-(2-bromo-4-methoxy-6-methylphenyl)-2,2,2-trifluoroacetamide (257b)

Compound 257b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-4-methoxy-6-methylaniline (257a) (5.00 g, 23.14 mmol; CAS # 1100394-71-

5) in DCM (25 mL) using triethylamine (3.98 g, 39.3 mmol), a solution of trifluoroacetic acid anhydride (7.29 g, 34.7 mmol) in DCM (5 mL) and stirring at RT for 1 h. This gave after workup N-(2-bromo-4-methoxy-6-methylphenyl)-2,2,2-trifluoroacetamide (257b) (7.05 g, 98% yield) as a plum solid; Ή NMR (300 MHz, DMSO-J₆) δ 11.02 (s, 1H), 7.17 (d,J= 2.8

Hz, 1H), 6.96 (d, 1H), 3.79 (s, 3H), 2.17 (s, 3H).

¹⁹F NMR (282 MHz, DMSO-</₆) δ -74.07; MS (ES-): 310/312 (M-l).

458

Step-2: Préparation of 2-amino-5-methoxy-7-methyl-lH-indole-3-carbonitrile (257c)

Compound 257c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-4-methoxy-6-methyIphenyl)-2,2,2-trifluoroacetamide (257b) (7.05 g, 22.59 mmol) in DMSO (20 mL) using malononitrile (1.791 g, 27.1 mmol), L-proline (0.520 g, 4.52 mmol), Cul (0.430 g, 2.259 mmol), a solution of K2CO3 (6.24 g, 45.2 mmol) in water (20 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [SiChgel (40 g), MeOH in DCM from 0-6%] 2-amino-5-methoxy-7-methyl-lHindole-3-carbonitrile (257c) (2.72 g, 60% yield) as a pale-brown solid; ’HNMR (300 MHz, DMSO-î/ô) δ 10.57 (s, 1H), 6.51 (d, J = 2.4 Hz, 1H), 6.43 (s, 2H), 6.35 (dd, J = 2.4, 0.9 Hz,

1H), 3.71 (s, 3H), 2.29 (s, 3H); MS (ES+): 202 (M+l), (ES-): 200 (M-l).

Step-3: Préparation of 6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-4-amine (257d)

Compound 257d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5-methoxy-7-methyl-lH-indole-3-carbonitrile (257c) (2.72 g, 13.52 mmol) using NH4OAC (3.13 g, 40.6 mmol), HC(OMe)3 (7.17 g, 67.6 mmol) and heating at 90 °C for

16 h. This gave after work up 6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-4-amine (257d) (2.97 g, 96% yield) as a pale-yellow solid; *H NMR (300 MHz, DMSO-d₆) δ 11.64 (s, 1H), 8.21 (s, 1H), 7.67 (d, J = 2.4 Hz, 1H), 7.13 (s, 2H), 6.79 (d, J = 2.3 Hz, 1H), 3.83 (s, 3H), 2.48 (s, 3H); MS (ES+): 229 (M+l), (ES-): 227 (M-l).

Step-4: Préparation of tert-butyl 2-(4-amino-6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol20 9-yl)acetate (257e)

Compound 257e was prepared according to the procedure reported in step-1 of scheme-1, from 6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-4-amine (257d) (2.00 g, 8.76 mmol) in DMF (15 mL) using tert-butyl 2-bromoacetate (1.880 g, 9.64 mmol), CS2CO3 (5.71 g, 17.52 mmol) and stirring at RT for 16 h. This gave after work up and purification using flash column chromatography [SiO₂ gel (40 g), methanol in DCM from 0-4%] tert-butyl 2-(4amino-6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (257e) (1.37 g, 46% yield) as a tan solid; Ή NMR (300 MHz, DMSO-d₆) δ 8.25 (s, 1H), 7.71 (d, J= 2.5 Hz, 1H), 7.30 (s, 2H), 6.79 (d, J= 2.4 Hz, 1H), 5.28 (s, 2H), 3.85 (s, 3H), 2.58 (s, 3H), 1.42 (s, 9H).

Step-5 : Préparation of 2-(4-amino-6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (257f)

I 21436 ! 459

Compound 257f was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (257e) | (1.00 g, 2.92 mmol) in DCM (10 mL) using TFA (3.33 g, 29.2 mmol) and stirring at RT for

h. This gave after work up TFA sait of 2-(4-amino-6-methoxy-8-methyl-9H- pyrimido[4,5-b]indoI-9-yl)acetic acid (257f) (1.51 g); Ή NMR (300 MHz, DMSO-t/e) δ 8.82 (s, 2H), 8.61 (s, 1H), 7.87 (d, J = 2.4 Hz, 1H), 6.97 (d, J = 2.4 Hz, 1H), 5.42 (s, 2H), 3.87 (s, 3H), 2.65 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-dd) δ -74.55; MS (ES+): 287 (M+l), (ES-): 285 (M-l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methoxy-8-methyl-9H-pyrimido[4,510 b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (257g)

Compound 257g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-methoxy-8-methyl-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (257f) (125 mg, 0.312 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(675 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (99 mg, 0.312 mmol), HATU (142 mg, 0.375 mmol), DIPEA (202 mg, 1.561 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (257g) (116 mg, 68% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-îZô) δ 10.82 (s, 1H, D₂O exchangeable), 8.69 (bs, 2H, D2O exchangeable), 8.58 (s, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.85 (d, J = 2.4 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 6.94 (d, J = 2.4 Hz,

1H), 5.88 (d, J = 17.9 Hz, 1H), 5.60 (d, J = 17.9 Hz, 1H), 4.42 (dd, J = 9.1, 5.7 Hz, 1H), 3.96

- 3.87 (m, 1H), 3.85 (s, 3H), 2.66 (s, 3H), 2.42 - 2.28 (m, 1H), 2.25 - 2.12 (m, 1H), 2.00 1.83 (m, 1H), 1.13 - 0.99 (m, 1H), 0.71 - 0.62 (m, 1H); MS (ES+): 550/552 (M+l), (ES-): 548/550 (M-l); Analysis calculated for C25H24BrN₇O3.HC1.2.5H₂O: C, 47.52; H, 4.79; Cl, 5.61; N, 15.52; Found: C, 47.55; H, 4.64; Cl, 5.61; N, 15.28.

Scheme 258

460

257f

258a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (258a)

Compound 258a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (2571) (125 mg, 0.312 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (104 mg, 0.312 mmol), HATU (142 mg, 0.375 mmol), DIPEA (202 mg, 1.561 mmol) and stirring at RT for

16 h. This gave after workup and purification by flash column chromatography [silica gel (12

g), MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (258a) (131 mg, 74% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ

10.83 (s, 1H, D2O exchangeable), 8.67 (bs, 2H, D2O exchangeable), 8.58 (s, 1H), 8.02 (d, J = 8.1 Hz, 1H), 7.85 (d, J = 2.4 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 6.94 (d, J = 2.3 Hz, 1H), 5.84 (d, J = 18.0 Hz, 1H), 5.57 (d, J = 17.9 Hz, 1H), 4.38 (dd, J = 9.0, 6.2 Hz, 1H), 3.86 (s, 3H), 3.72 - 3.66 (m, 1H), 2.65 (s, 3H), 2.49 - 2.39 (m, 1H), 1.98 (m, 1H),

1.31 (s, 3H), 1.01 (m, 1H), 0.83 (m, 1H); MS (ES+): 564/566 (M+l), (ES-): 562/564 (M-l);

Analysis calculated for C₂6H26BrN₇O3.1.2HC1.2.5H₂O: C, 47.81; H, 4.97; Cl, 6.51; N, 15.01; Found: C, 47.85; H, 4.89; Cl, 6.51; N, 14.94.

Scheme 259

461

259b

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(2-bromopyridin-4-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (259d)

Step-1: Préparation of (lR,3S,5R)-tert-butyl 3-((2-bromopyridin-4-yl)carbamoyl)-25 azabicyclo[3.1.0]hexane-2-carboxylate (259b)

Compound 259b was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (259a) (0.5 g, 2.200 mmol; CAS # 197142-34-0) in DCM (15 mL) using 1-methyl-lHimidazole (0.438 mL, 5.50 mmol), methanesulfonyl chloride (0.204 mL, 2.64 mmol), 210 bromopyridin-4-amine (0.381 g, 2.200 mmol; CAS # 7598-35-8) and stirring at RT for 18 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), EtOAc in hexane from 0 - 50%] (lR,3S,5R)-tert-butyl 3-((2-bromopyridin-4-yl)carbamoyl)2-azabicyclo[3.1.0]hexane-2-carboxylate (259b) (0.798 g, 95% yield) as a colorless gum; MS (ES+): 382.1 (M+l).

Step-2: Préparation of (lR,3S,5R)-N-(2-bromopyridin-4-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (259c)

Compound 259c was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-tert-butyl 3-((2-bromopyridin-4-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane2-carboxylate (259b) (0.78 g, 2.041 mmol) in DCM (11 mL) using TFA (1.100 mL, 14.28

462 mmol) and stirring overnight at RT. This gave after work up and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCI) from 0-100%] HCl sait of (lR,3S,5R)-N-(2-bromopyridin-4-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (259c) (450 mg, 69% yield); 'H NMR (300 MHz,

DMSO-cfc) δ 11.53 (s, 1H), 10.11 (s, 1H), 9.28 (s, 1H), 8.32 (d, 5.5 Hz, 1H), 7.92 (d, J =

1.8 Hz, 1H), 7.60 (dd, J= 5.6, 1.9 Hz, 1H), 4.24 (t, J= 9.4 Hz, 1H), 3.45 - 3.29 (m, 1H), 2.65 (dd, J= 12.8, 7.7 Hz, 1H), 2.11 (td, J= 12.0, 4.8 Hz, 1H), 1.84 (dq, J= 9.9, 5.1 Hz, 1H), 0.91 - 0.79 (m, 2H); MS (ES+): 282.0 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,510 b]indol-9-yl)acetyl)-N-(2-bromopyridin-4-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (259d) ·

Compound 259d was prepared according to the procedure reported in step-3 of scheme-1, from HCI sait of (lR,3S,5R)-N-(2-bromopyridin-4-yI)-2-azabicyclo[3.1.0]hexane-3carboxamide (259c) (55.1 mg, 0.173 mmol) in DMF (1.5 mL) using HCl sait of 2-(4-amino15 6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (60mg, 0.173 mmol), HATU (99 mg, 0.260 mmol), DIPEA (0.151 mL, 0.865 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 -100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1%

HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(trifhioromethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(2-bromopyridin-4-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (259d) (54 mg, 54% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î7₆) δ 10.59 (s, 1H, D₂O exchangeable), 8.99 (s, 1H), 8.79 - 8.53 (m, 3H, 2H D2O exchangeable), 8.20 (d, J= 5.6 Hz, 1H), 7.96 - 7.80 (m, 3H), 7.47 (dd, J= 5.6, 1.9 Hz, 1H), 5.84 (d, J= 17.4 Hz, 1H), 5.50 (d, J= 17.3 Hz, 1H), 4.28 (dd, J= 9.0, 6.0 Hz, 1H), 4.02 - 3.89 (m, 1H), 2.38 (dd, J= 13.4,

9.2 Hz, 1H), 2.29-2.15 (m, 1H), 2.00-1.86 (m, 1H), 1.16-1.00 (m, 1H), 0.92-0.64 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-t/₆) δ -58.55. MS (ES+): 574.1/576.1 (M+l); (ES-): 572.1/574.1 (M-l); Analysis calculated for C24Hi₉BrF3N₇O₂ 2.5H₂O.1.25HC1: C, 43.35; H, 3.83; Cl, 6.66; N, 14.74; Found: C, 43.46; H, 3.57; Cl, 6.83; N, 14.67.

Scheme 260

463

CF₃

260a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyrazin-2-yl)-2-azabicycIo[3.1.0]hexane-3-carboxamide (260a)

Compound 260a was prepared according to the procedure reported in step-3 of scheme-1, 5 from HCl sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (60mg, 0.173 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (112a) (55.3 mg, 0.173 mmol), HATU (99 mg, 0.260 mmol), DIPEA (0.151 mL, 0.865 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 10 g), eluting with DMA-80 in DCM from 0 -100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%](lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (260a) (37 mg, 37% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-ùfe) δ 11.14 15 (s, 1H, D₂O exchangeable), 9.24 (s, 1H), 8.98 (s, 1H), 8.68 - 8.56 (m, 3H, 2H D₂O exchangeable), 8.54 (s, 1H), 7.94 - 7.82 (m, 2H), 5.83 (d, J= 17.4 Hz, 1H), 5.48 (d, J= 17.3 Hz, 1H), 4.44 (dd, J= 9.0, 5.6 Hz, 1H), 3.93 (td, J= 6.5, 5.7, 2.3 Hz, 1H), 2.42 - 2.20 (m, 2H), 2.00-1.86 (m, 1H), 1.12-1.01 (m, 1H), 0.90 - 0.74 (m, 1H). ^I9F NMR (282 MHz, DMSO) δ -58.53. MS (ES+): 575.1/577.1 (M+l); (ES-): 573.1/575.1 (M-l); Analysis calculated for C^HisBrFsNsCh 2.5H₂O.1.15HC1.:C, 41.71; H, 3.68; Cl, 6.16; N, 16.92;

Found: C, 41.73; H, 3.45; Cl, 5.90; N, 16.76.

Scheme 261

Ί

464

Ο ιι S. II Ο

261b

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (261d)

Step-1 : Préparation of (lR,3S,5R)-teri-butyl 3-((6-bromopyrazin-2-yl)carbamoyl)-5-methyl2-azabicyclo[3.1.0]hexane-2-carboxylate (261b)

Compound 261b was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(terZ-butoxycarbonyl)-5-methyl-2-azabicyclo[3.1,0]hexane-3-carboxylic acid (261a) (0.53 g, 2.197 mmol; CAS # 1306734-44-0) in DCM (15 mL) using 1-methyl10 lH-imidazole (0.438 mL, 5.49 mmol), methanesulfonyl chloride (0.204 mL, 2.64 mmol) and 6-bromopyrazin-2-amine (0.382 g, 2.197 mmol; CAS # 54237-53-5) and stirring at RT for 18 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with EtOAc in hexane from 0 - 50%] (lR,3S,5R)-tert-butyl 3-((6-bromopyrazin-2yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (261b) (0.418 g, 48% yield) as a colorless gum; MS (ES+): 397.1/399.1 (M+l).

Step-2: Préparation of (lR,3S,5R)-N-(6-bromopyrazin-2-yl)-5-methyI-2azabicyclo[3.1.0]hexane-3-carboxamide (261c)

Compound 261c was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-fërZ-butyl 3-((6-bromopyrazin-2-yl)carbamoyl)-5-methyl-2• ·

465 azabicyclo[3.1.0]hexane-2-carboxylate (261b) (0.41 g, 1.032 mmol) in DCM (6 mL) using TFA (0.557 mL, 7.22 mmol) and stirring ovemight at RT. This gave after work up and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCI) from 0-100%] HCl sait of (lR,3S,5R)-N-(65 bromopyrazin-2-yI)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (261c) (232 mg, 67% yield); Ή NMR (300 MHz, DMSO-îZ₆) δ 11.70 (s, 1H), 9.67 (s, 2H), 9.27 (s, 1H), 8.67 (s, 1H), 4.23 (dd, J= 11.0, 7.7 Hz, 1H), 3.10 (dd, J= 6.9, 2.4 Hz, 1H), 2.65 (dd, J= 12.7, 7.7 Hz, 1H), 2.00 (t, J= 11.8 Hz, 1H), 1.25 (s, 3H), 1.02 (dd, J= 7.2, 2.4 Hz, 1H), 0.78 (t, J= 7.0 Hz, 1H); MS (ES+): 297.0 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyI)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (261d)

Compound 261d was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (lR,3S,5R)-N-(6-bromopyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane15 3-carboxamide (261c) (58 mg, 0.173 mmol) in DMF (1.5 mL) using HCl sait of 2-(4-amino6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (60mg, 0.173 mmol), HATU (99 mg, 0.260 mmol), DIPEA (0.151 mL, 0.865 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (261d) (62 mg, 61% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-dô) δ 11.14 (s, 1H, D₂O exchangeable), 9.25 (s, 1H), 8.95 (s, 1H), 8.56 (s, 1H), 8.54 (s, 1H), 8.40 (s, 2H,

D₂O exchangeable), 7.88 - 7.80 (m, 2H), 5.76 (d, J= 17.4 Hz, 1H), 5.42 (d, J= 17.3 Hz, 1H), 4.39 (dd, J= 9.1, 6.0 Hz, 1H), 3.71 (dd, J= 5.5, 2.5 Hz, 1H), 2.48 - 2.42 (m, 1H), 2.03 (dd, J = 13.3, 6.0 Hz, 1H), 1.32 (s, 3H), 1.05 - 0.91 (m, 2H). ¹⁹F NMR (282 MHz, DMSO-îZ₆) δ 58.48; MS (ES+): 589.1/591.1 (M+l); (ES-): 587.1/589.1 (M-l); Analysis calculated for C₂₄H₂₀BrF₃N₈O₂ 1.75H₂O.1.1HC1.:C, 43.61; H, 3.75; Cl, 5.90; N, 16.95; Found: C, 43.54; H,

3.74; Cl, 5.59; N, 16.70.

Scheme 262

466

^215f 262a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(dimethylamino)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (262a)

Compound 262a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(dimethylamino)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (215f) (75 mg, 0.188 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (59.8 mg, 0.188 mmol), HATU (86 mg, 0.225 mmol), DIPEA (121 mg, 0.939 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(dimethylamino)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (262a) (13 mg, 13% yield) as a pale-yellow solid; ’H NMR (300 MHz, DMSO-î7<î) δ 10.77 (s, 1H, D₂O exchangeable), 9.08 - 8.52 (m, 4H, 2H D₂O exchangeable), 8.01 (d, J = 8.2 Hz, 1H), 7.88 7.55 (m, 3H), 7.32 (d, J = 7.7 Hz, 1H), 5.77 (d, J = 17.4 Hz, 1H), 5.42 (d, J = 17.2 Hz, 1H), 4.41 (dd, J = 9.1, 5.6 Hz, 1H), 3.91 (td, J = 6.4, 5.6, 2.4 Hz, lH),3.18(s, 6H), 2.41-2.28 (m, 1H), 2.27-2.14 (m, 1H), 1.99-1.84 (m, 1H), 1.13-1.02 (m, 1H), 0.84-0.73 (m, 1H); MS (ES+): 549/551 (M+l); (ES-): 547/549 (M-l).

Scheme 263

467

192e 263a

263b 263c

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(pyridin-4-yl)-9H-pyrimido[4,5b]indol-9-yl)acetyI)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (263c)

Step-1 : Préparation of tert-butyl 2-(4-amino-8-methyl-6-(pyridin-4-yl)-9H-pyrimido[4,5b]indol-9-yl)acetate (263a)

To a suspension of tert-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (192e) (300 mg, 0.767 mmol), pyridin-4-yIboronic acid (141 mg, 1.150 mmol), Pd2(dba)s (70.2 mg, 0.077 mmol), XPhos (73.1 mg, 0.153 mmol), and 1.27 M aqueous K3PO4 (1.207 mL, 1.533 mmol) in 10 n-BuOH (4 mL) in a scintillation vial. The vial was flushed with nitrogen and then heated at 100 °C under nitrogen for 16 h. The resulting cooled deep red mixture was concentrated to dryness and the residue was suspended in H₂O (30 mL) and EtOAc (30 mL), stirred for 30 min, and then fîltered. The fîltered cake was washed with H₂O (20 mL) and EtOAc (20 mL) to provide 263a as a pale-yellow solid. The filtrate separated from aqueous layer and the aqueous layer was extracted with EtOAc (25 mL). The combined organic extract was washed with H2O (25 mL), brine (25 mL), dried, fîltered and concentrated, and residue obtained was purified by flash column chromatography (Silica gel, 12 g, 0-7% MeOH in DCM) to provide additional 263a as a white solid. Total yield: ZerZ-butyl 2-(4amino-8-methyl-6-(pyridin-4-yI)-9H-pyrimido[4,5-b]indol-9-yl)acetate (263a) (250 mg, 84 % yield). 'H NMR (300 MHz, DMSO-d₆) δ 8.76 - 8.55 (m, 3H), 8.33 (s, 1H), 8.01 - 7.88 (m, 2H), 7.69 (s, 1H),

468

7.52 (s, 2H), 5.38 (s, 2H), 2.72 (s, 3H), 1.44 (s, 9H). LC-MS: t = 1.73 min; MS (ES+) 390 (M+l), (ES-) 388 (M-l).

Step-2: Préparation of 2-(4-amino-8-methyl-6-(pyridin-4-yl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (263b)

Compound 263b was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-8-methyl-6-(pyridin-4-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (263a) (250 mg, 0.642 mmol) in DCM (5 mL) using TFA (732 mg, 6.42 mmol) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-8-methyl-6-(pyridin-4-yl)9H-pyrimido[4,5-b]indol-9-yl)acetic acid (263b) (394 mg); Ή NMR (300 MHz, DMSO-î/₆) δ

8.94 (d, J = 6.1 Hz, 2H), 8.89 (s, 1H), 8.60 - 8.44 (m, 3H), 8.19 (s, 2H), 7.99 (s, 1H), 5.46 (s,

2H), 2.79 (s, 3H). ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -74.43;

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(pyridin-4-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-315 carboxamide (263c)

Compound 263c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(pyridin-4-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (263b) (56 mg, 0.125 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (39.9 mg, 0.125 mmol),

HATU (57.1 mg, 0.150 mmol), DIPEA (81 mg, 0.626 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 8%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(pyridin-4-yl)-9H-pyrimido[4,525 b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (263c) (32 mg, 43% yield) HCl sait as a pale-yellow solid; ’H NMR (300 MHz, DMSO-î/ô) δ 10.84 (s, 1H, D₂O exchangeable), 9.02 (s, 1H), 8.96 (d, J= 6.5 Hz, 2H), 8.83 (s, 2H, D2O exchangeable), 8.68 - 8.57 (m, 3H), 8.09 - 7.92 (m, 2H), 7.70 (t, J= 8.0 Hz, 1H), 7.30 (d, J= 7.7 Hz, 1H), 5.97 (d, J= 18.0 Hz, 1H), 5.67 (d, J= 17.9 Hz, 1H), 4.43 (dd, J= 9.0, 5.7 Hz,

1H), 3.99-3.90 (m, 1H),2.81 (s, 3H), 2.42 - 2.29 (m, 1H), 2.28 - 2.14 (m, 1H), 2.02 - 1.85 (m, 1H), 1.17-1.03 (m, 1H), 0.79 - 0.59 (m, 1H); MS (ES+): 597/599 (M+l), (ES-): 595/597 (M-l); Analysis calculated for C29H25BrN8O2.1.9HC1.4.25H2O: C, 46.86; H, 4.80; Cl, 9.06; N, 15.08; Found: C, 46.75; H, 4.43; Cl, 9.04; N, 14.85.

469

Scheme 264

263b 264a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(pyridin-4-yl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-35 carboxamide (264a)

Compound 264a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(pyridin-4-yI)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (263b) (75 mg, 0.168 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicycIo[3.1.0]hexane-3-carboxamide (8a) (55.8 mg, 0.168 mmol), HATU (76 mg, 0.201 mmol), DIPEA (108 mg, 0.838 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 8%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(pyridin-4-yl)-9H-pyrimido[4,515 b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (264a) (69 mg, 67% yield) HCl sait as a pale-yellow solid; 'H NMR (300 MHz, DMSO-dô) δ 10.85 (s, 1H, D₂O exchangeable), 9.11 (bs, 2H, D2O exchangeable), 9.01 (s, 1H), 8.94 (d, J = 6.3 Hz, 2H), 8.73 - 8.55 (m, 3H), 8.01 (d, J= 8.2 Hz, 1H), 7.96 (s, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.30 (d, J = 7.7 Hz, 1H), 5.93 (d, J = 18.0 Hz, 1H), 5.59 (d, J = 17.9 Hz,

1H), 4.40 (dd, J = 9.0, 6.2 Hz, 1H), 3.75 (m, 1H), 2.78 (s, 3H), 2.50 - 2.44 (m, 1H), 1.99 (m,

1H), 1.33 (s, 3H), 1.15 - 0.96 (m, 1H), 0.86 (m, 1H); MS (ES+): 611/613 (M+l), (ES-): 609/611 (M-l).

Scheme 265

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-hydroxy-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (265a)

Compound 265a was prepared according to the procedure reported in scheme-164, from (lR,3S,5R)-2-(2-(4-amino-6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (258a) (78 mg, 0.138 mmol) in DCM (5 mL) using a solution of 1.0 M BBn in DCM (0.691 mL, 0.691 mmol) and stirring at RT for 16 h. This gave after work up and purification using reverse phase column chromatography [C 18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-hydroxy-8-methyl-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (265a) (30 mg, 36% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 10.89 and 10.82 (2s, 1H, D₂O exchangeable), 9.39 (s, 1H, D2O exchangeable), 8.77 - 8.41 (m, 3H, 2H D2O exchangeable), 8.01 and 7.94 (2d, J = 8.2 Hz, 1H), 7.77 - 7.62 (m, 2H), 7.35 - 7.27 (m, 1H), 6.82 (s, 1H), 5.80 (d, J= 17.9 Hz, 1H), 5.53 (d, J= 17.8 Hz, 1H), 4.38 (dd, J= 9.1, 6.2 Hz, 1H), 3.72 - 3.63 (m, 1H), 2.65 and 2.60 (2s, 3H), 2.48 - 2.40 (m, 1H), 1.97 (m, 1H), 1.30 (s, 3H), 1.00 (m, 1H), 0.91 - 0.76 (m, 1H); MS (ES+): 550/552 (M+l), (ES-): 548/550 (M-l); Analysis calculated for

C25H24BrN7O3.Li5HCl.2H2O: C, 47.79; H, 4.68; Cl, 6.49; N, 15.60; Found: C, 47.69; H, 4.42; Cl, 6.41; N, 15.42.

Scheme 266

471

266b

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(3-bromophenyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (266b)

Compound 266b was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (60mg, 0.173 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(3bromophenyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (266a) [(55.0 mg, 0.173 mmol); prepared according to the procedure reported by Lorthiois, E. et al. in J. Med. Chem. (2017), 60(13), 5717-5735.], HATU (99 mg, 0.260 mmol), DIPEA (0.151 mL, 0.865 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(3-bromophenyl)-2- azabicyclo[3.1.0]hexane-3-carboxamide (266b) (73 mg, 74% yield) as a white solid; *H NMR (300 MHz, DMSO-J₆) δ 10.09 (s, 1H, D₂O exchangeable), 8.99 (s, 1H), 8.65 - 8.45 (m, 3H, 2H D₂O exchangeable), 7.97 - 7.81 (m, 3H), 7.45 (dt, J= 6.9, 2.3 Hz, 1H), 7.28 7.18 (m, 2H), 5.82 (d, J= 17.4 Hz, 1H), 5.50 (d, J= 17.3 Hz, 1H), 4.27 (dd, J= 9.0, 5.8 Hz, 1H), 3.99-3.87 (m, 1H), 2.42-2.28 (m, 1H), 2.28-2.17 (m, 1H), 1.99- 1.86 (m, 1H), 1.14

- 1.02 (m, 1H), 0.83 - 0.74 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-d₆) δ -58.53; MS (ES+):

573.1/575.1 (M+l); (ES-): 571.1/573.1 (M-l); Analysis calculated for C₂5H₂oBrF₃N₆0₂.1.25H₂O.HCl: C, 47.48; H, 3.75; Cl, 5.61; N, 13.29; Found: C, 47.47; H, 3.43; Cl, 5.35; N, 13.23.

Scheme 267

267c

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-chloropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (267c)

Step-1: Préparation of (IR,3S,5R)-ZerZ-butyl 3-((6-chloropyridin-2-yl)carbamoyl)-25 azabicyclo[3.1.0]hexane-2-carboxy!ate (267a)

Compound 267a was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(ZerZ-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (259a) (0.5 g, 2.200 mmol) in DCM (15 mL) using 1-methyl-lH-imidazole (0.438 mL, 5.50 mmol), methanesulfonyl chloride (0.204 mL, 2.64 mmol), 6-chloropyridin-2-amine (0.283 g, 10 2.200 mmol; CAS # 45644-21-1) and stirring at RT for 18 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with EtOAc in hexane from 0 - 50%] (lR,3S,5R)-ZerZ-butyl 3-((6-chloropyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (267a) (0.580 g, 78% yield) as a white solid; ’H NMR (300 MHz, DMSO-cfc) δ 10.80 (s, 1H), 8.11 - 8.01 (m, 1H), 7.86 (t, J= 8.0 Hz, 1H), 7.22 (d, 15 1H), 4.20-4.00 (m, 1H), 3.44 - 3.37 (m, 1H), 2.30 (dd, J= 13.2, 8.8 Hz, 1H),

2.21-2.01 (m, 1H), 1.59 (s, 1H), 1.45-1.08 (m, 9H), 0.80-0.65 (m, 1H), 0.48-0.27 (m, 1H); MS (ES+): 338.1(M+1).

Step-2: Préparation of (lR,3S,5R)-N-(6-chloropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (267b)

473

Compound 267b was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-/er/-butyl 3-((6-chloropyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane2-carboxylate (267a) (0.565 g, 1.673 mmol) in DCM (9 mL) using TFA (0.902 mL, 11.71 mmol) and stirring ovemight at RT. This gave after work up and purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] HCl sait of (lR,3S,5R)-N-(6-chloropyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (267b) (413 mg, 90% yield); 'HNMR (300 MHz, DMSO-ί/ό) δ 11.35 (s, 1H), 10.01 (s, 1H), 9.23 (s, 1H), 8.06-7.88 (m, 2H), 7.33 (dd, J= 7.6, 0.9 Hz, 1H), 4.21-4.08 (m, 1H), 3.39 - 3.27 (m, 1H), 2.62 (dd, J= 12.8, 7.7 Hz, 1H), 2.1510 2.02 (m, 1H), 1.89 - 1.76 (m, 1H), 0.90 - 0.76 (m, 2H); MS (ES+): 238.1 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyI)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (267c) .

Compound 267c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (lR,3S,5R)-N-(6-chloropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (267b) (47.4 mg, 0.173 mmol) in DMF (1.5 mL) using HCl sait of 2-(4-amino6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (60 mg, 0.173 mmol), HATU (99 mg, 0.260 mmol), DIPEA (0.151 mL, 0.865 mmol) and stirring at RT for 16 h.

This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (267c) (67 mg, 73% yield) as a white solid; ’H NMR (300 MHz, DMSO-î/ô) δ 10.75 (s, 1H,

D2O exchangeable), 8.98 (s, 1H), 8.61 (s, 1H), 8.58 (s, 2H, D₂O exchangeable), 7.98 (d, J= 8.2 Hz, 1H), 7.89 - 7.76 (m, 3H), 7.19 (d, J= 7.7 Hz, 1H), 5.82 (d, J= 17.4 Hz, 1H), 5.47 (d, J= 17.3 Hz, 1H), 4.42 (dd, J= 9.1, 5.5 Hz, 1H), 3.92 (td, J= 6.2, 5.4, 2.3 Hz, 1H), 2.40 2.15 (m, 2H), 2.00 - 1.83 (m, 1H), 1.15 - 0.98 (m, 1H), 0.86 - 0.74 (m, 1H); ¹⁹F NMR (282

MHz, DMSO-cfc) δ -58.53. MS (ES+): 530.2 (M+l); (ES-): 528.1 (M-l); Analysis calculated for C₂4Hi9ClF3N₇O₂.2H2O.0.85HCl: C, 48.29; H, 4.03; Cl, 10.99; N, 16.43; Found: C, 48.33; H, 3.79; Cl, 10.82; N, 16.28

Scheme 268

474

11e 268b

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(3-bromo-2-fluorophenyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (268b)

Compound 268b was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of 2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (60mg, 0.173 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(3-bromo-2fluorophenyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (268a) [(58.1 mg, 0.173 mmol); prepared according to the procedure reported by Lorthiois, E. et al. J. Med. Chem. (2017), 60(13), 5717-5735.], HATU (99 mg, 0.260 mmol) and DIPEA (0.151 mL, 0.865 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with DMA80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indoI-9-yl)acetyl)-N-(3-bromo-2-fluorophenyl)-215 azabicyclo[3.1,0]hexane-3-carboxamide (268b) (98 mg, 96% yield) as a white solid; *H NMR (300 MHz, DMSO-dô) δ 9.85 (s, 1H, D₂O exchangeable), 8.96 (s, 1H), 8.55 (s, 1H), 8.36 (s, 2H, D₂O exchangeable); 7.90 - 7.78 (m, 2H), 7.78 - 7.67 (m, 1H), 7.49 - 7.38 (m, 1H), 7.11 (td, J= 8.1, 1.4 Hz, 1H), 5.77 (d, J= 17.5 Hz, 1H), 5.49 (d, J= 17.3 Hz, 1H), 4.46 (dd, J= 8.8, 5.5 Hz, 1H), 3.94-3.84 (m, 1H), 2.38-2.22 (m, 2H), 2.00-1.84 (m, 1H), 1.10 (m,

1H), 0.86 - 0.71 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î7₆) δ -58.45, -117.67; MS (ES+):

591.1/593.1 (M+l); (ES-): 589.1/591.1 (M-l); Analysis calculated for C₂₅Hi9BrF4N₆O₂.1.5H₂O.0.95HCl :C, 45.98; H, 3.54; Cl, 5.16; N, 12.87; Found: C, 45.92; H, 3.41; Cl, 4.94; N, 12.73.

Scheme 269

475

269b

269c

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyI)-N-(6-chloropyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (269c)

Step-1 : Préparation of (1 R,3S,5R)-ZerZ-butyl 3-((6-chloropyridin-2-yl)carbamoyl)-5-methyl2-azabicyclo[3.1.0]hexane-2-carboxylate (269a)

Compound 269a was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(ieri-butoxycarbonyl)-5-methyI-2-azabicycIo[3.1.0]hexane-3-carboxylic acid (261a) (0.531 g, 2.200 mmol) in DCM (15 mL) using 1-methyl-IH-imidazole (0.438 mL, 5.50 mmol), methanesulfonyl chloride (0.204 mL, 2.64 mmol), 6-chloropyridin-2-amine (0.283 g, 2.200 mmol; CAS # 45644-21-1) and stirring at RT for 18 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with EtOAc in hexane from 0 - 50%] (lR,3S,5R)-terZ-butyl 3-((6-chloropyridin-2-yl)carbamoyl)5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (269a) (0.667 g, 86% yield) as a white solid; *H NMR (300 MHz, DMSO-î/₆) δ 10.80 (s, 1H), 8.16 - 7.96 (m, 1H), 7.86 (t, J= 7.9 Hz, 1H), 7.22 (d, J= 7.7 Hz, 1H), 4.19-4.01 (m, 1H), 3.23 - 3.06 (m, 1H), 2.39 (dd, J= 12.8, 8.7 Hz, 1H), 1.95 - 1.81 (m, 1H), 1.51 - 1.08 (m, 12H), 0.71-0.62 (m, 1H), 0.620.52 (m, 1H); MS (ES+): 352.1(M+1).

Step-2: Préparation of (lR,3S,5R)-N-(6-chloropyridin-2-yl)-5-methyl-220 azabicyclo[3.1.0]hexane-3-carboxamide (269b)

476

Compound 269b was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-tert-butyl 3-((6-chloropyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexane-2-carboxylate (269a) (0.652 g, 1.853 mmol) in DCM (10 mL) using TFA (0.999 mL, 12.97 mmol) and stirring ovemight at RT. This gave after work up and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] HCl sait of (lR,3S,5R)-N-(6chloropyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (269b) (532 mg, 100% yield); Ή NMR (300 MHz, DMSO-J₆) δ 11.34 (s, 1H), 9.85 (s, 1H), 9.24 (s, 1H), 8.08 - 7.85 (m, 2H), 7.40 - 7.21 (m, 1H), 4.27 - 4.08 (m, 1H), 3.09 (dd, J= 7.0, 2.4 Hz, 1H), 2.65 (dd, J= 12.6, 7.6 Hz, 1H), 1.97 (t, J= 11.9 Hz, 1H), 1.25 (s, 3H), 1.05-0.94 (m, 1H), 0.82 0.72 (m, 1H); MS (ES+): 252.1 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yI)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (269c)

Compound 269c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (lR,3S,5R)-N-(6-chloropyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane3-carboxamide (269b) (60 mg, 0.173 mmol) in DMF (1.5 mL) using HCl sait of 2-(4-amino6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) [(58.1 mg, 0.173 mmol), HATU (99 mg, 0.260 mmol), DIPEA (0.151 mL, 0.865 mmol) and stirring at RT for 16 h.

This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-325 carboxamide (269c) (53 mg, 56% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-îZô) δ 10.75 (s, 1H, D2O exchangeable), 8.98 (s, 1H), 8.70 - 8.47 (m, 3H, 2H D2O exchangeable), 7.99 (d, J= 8.2 Hz, 1H), 7.90 - 7.75 (m, 3H), 7.19 (d, J= 7.7 Hz, 1H), 5.78 (d, J= 17.4 Hz, 1H), 5.43 (d, J= 17.3 Hz, 1H), 4.37 (dd, J= 9.1, 6.0 Hz, 1H), 3.70 (m, 1H), 2.49-2.42 (m, 1H), 1.99 (m, 1H), 1.32 (s, 3H), 1.07 - 0.98 (m, 1H), 0.98 - 0.90 (m, 1H).

MS (ES+): 544.2 (M+l); (ES-): 542.1 (M-l); Analysis calculated for C25H21CIF3N7O2.2H2O.O.75HCI: C, 49.44; H, 4.27; Cl, 10.22; N, 16.14; Found: C, 49.65; H, 4.00; Cl, 9.94; N, 16.05.

Scheme 270

477

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N(6-bromopyridin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (270a)

Compound 270a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (62b) (90 mg, 0.283 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (8a) (113 mg, 0.339 mmol), HATU (161 mg, 0.424 mmol), DIPEA (0.197 mL, 1.131 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in

DCM from 0 - 5%] followed by purification using reverse phase C18 column chromatography with water (containing 0.1% HCl)/acetonitrile (1:0 to 0:1) (lR,3S,5R)-2-(2(4-amino-6-phenyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl2-azabicyclo[3.1.0]hexane-3-carboxamide (270a) (58 mg, 34% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-cfe) δ 10.78 (s, 1H), 8.80 (d, J= 1.6 Hz, 1H), 8.76 (s, 2H),

8.63 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.92 - 7.75 (m, 4H), 7.69 (t, J= 8.0 Hz, 1H), 7.56 7.47 (m, 2H), 7.43 - 7.35 (m, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.76 (d, J= 17.3 Hz, 1H), 5.40 (d, J= 17.3 Hz, 1H), 4.38 (dd, J= 9.1, 6.0 Hz, 1H), 3.71 (dd, J= 5.5, 2.4 Hz, 1H), 2.58 2.41 (m, 1H), 1.99 (dd, J= 13.2, 5.9 Hz, 1H), 1.31 (s, 3H), 1.06 - 0.98 (m, 1H), 0.94 (m, 1H); MS (ES+): 596.15 & 598.15 (M+l), MS (ES-): 594.10 & 596.10 (M-l); Analysis calculated for C30H26BrN7O2.LlHCl.2H2O: C, 53.57; H, 4.66; N, 14.58; Cl, 5.80. Found: C, 53.76; H, 4.48; N, 14.50.; Cl, 5.76

Scheme 271

478

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(m-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (271c)

Step-1: Préparation of ethyl 2-(4-amino-6-(m-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (271a)

Compound 271a was prepared according to the procedure reported in step-1 of scheme-62, from ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (200 mg, 0.573 mmol) in dioxane (18 mL) using m-tolylboronic acid (117 mg, 0.859 mmol), a solution of césium carbonate (280 mg, 0.859 mmol) in water (2.2 mL), Pd(PPh.3)2C12 (80 mg, 0.115 mmol) and heating at 100 °C for 19 h under nitrogen. This gave after workup a mixture of ethyl 2-(4-amino-6-(m-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (271a) and 2-(4-amino-6m-tolyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (271b) which was used as such for the next step; MS (ES+): 361.10 & 333.15 (M+l).

Step-2: Préparation of 2-(4-amino-6-(m-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (271b)

Compound 271b was prepared according to the procedure reported in step-4 of scheme-17, from a mixture containing ethyl 2-(4-amino-6-(m-tolyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (271a) and 2-(4-amino-6-(m-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid

479 (271b) (207 mg, 0.573 mmol) in THF (5 mL) and methanol (5 mL) using a solution of lithium hydroxide hydrate (147 mg, 3.44 mmol) in water (5 mL) and stirring at RT for 21 h. The residue obtained after work up was used as such for the next step; MS (ES+): 333.10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(m-toIyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (271c)

Compound 271c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(m-toIyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (271b) (0.090 mg, 0.271 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-210 azabicyclo[3.1.0]hexane-3-carboxamide (4a) (0.104 g, 0.325 mmol), HATU (0.154 g, 0.406 mmol), DIPEA (140 mg, 1.083 mmol) and stirring at RT for 19 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [C18 column, eluting with ACN in water (containing 0.1% HCl) from 0-100%] (1R,3S,5R)75 2-(2-(4-amino-6-(m-tolyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (271c) (58 mg, 36% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î7₆) δ 10.78 (s, 1H), 8.84 - 8.70 (m, 3H), 8.64 (s, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.87 (dd, J= 8.7, 1.6 Hz, 1H), 7.81 - 7.59 (m, 4H), 7.39 (t, J= Ί.6 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 7.20 (d, J= 7.5 Hz, 1H), 5.80 (d, J= 17.4 Hz, 1H), 5.45 (d, J= 17.3

Hz, 1H), 4.42 (dd, J= 9.0, 5.6 Hz, 1H), 3.98 - 3.89 (m, 1H), 2.42 (s, 3H), 2.41 - 2.14 (m, 2H), 2.02 - 1.82 (m, 1H), 1.16 - 0.99 (m, 1H), 0.86 - 0.72 (m, 1H); MS (ES+): 596.20 & 598.10 (M+l), (ES-): 594.10 & 596.20 (M-l); Analysis calculated for C₃oH₂₆BrN₇0₂.HC1.2.25H₂0: C, 53.50; H, 4.71; N, 14.56; Cl, 5.26; Found: C, 53.79; H, 4.62; N, 14.71.; Cl, 5.19.

Scheme 272

480

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(pyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (272c)

Step-1: Préparation of (lR,3S,5R)-tert-butyl 3-(pyridin-2-ylcarbamoyl)-25 azabicyclo[3.1.0]hexane-2-carboxylate (272a)

Compound 272a was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(/erLbutoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (259a) (2.415 g, 10.63 mmol) in DCM (10 mL) using 1-methyl-IH-imidazole (2.181 g, 26.6 mmol), methanesulfonyl chloride (1.461 g, 12.75 mmol) in DCM (2 mL), pyridin-2-amine (1.00 g, 10.63 mmol; CAS # 504-29-0) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (80 g), eluting with EtOAc in hexane from 0 - 30%] (lR,3S,5R)-terZ-butyl 3-(pyridin-2-ylcarbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (272a) (2.54 g, 79% yield) as a sticky white solid; ’H NMR (300 MHz, DMSO-î7₆) δ 10.39 (s, 1H), 8.35 - 8.22 (m, 1H), 8.08 (d, J= 8.4 Hz, 1H),

7.87 - 7.73 (m, 1H), 7.16 - 7.03 (m, 1H), 4.24 - 4.07 (m, 1H), 3.52 - 3.35 (m, 1H), 2.29 (dd,

J= 13.2, 8.8 Hz, 1H), 2.18-2.04 (m, 1H), 1.64-1.51 (m, 1H), 1.32 (2bs, 9H), 0.82-0.70 (m, 1H), 0.43 - 0.36 (m, 1H); MS (ES+): 304.20 (M+l).

Step-2: Préparation of (lR,3S,5R)-N-(pyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (272b)

481

Compound 272b was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-terZ-butyl 3-(pyridin-2-ylcarbamoyl)-2-azabicyclo[3.1.0]hexane-2carboxylate (272a) (2.5 g, 8.24 mmol) in DCM (30 mL) using TFA (3.15 mL, 41.2 mmol) and stirring for 16 h at RT. This gave after work up TFA sait of (lR,3S,5R)-N-(pyridin-2-yl)5 2-azabicyclo[3.1.0]hexane-3-carboxamide (272b) (6.0 g); MS (ES+): 204.10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(pyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (272c)

Compound 272c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (lR,3S,5R)-N-(pyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (272b) (0.127 mg, 0.4 mmol) in DMF (10 mL) using HCl sait of 2-(4-amino-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (lie) (69.3 mg, 0.2 mmol), HATU (152 mg, 0.400 mmol), DIPEA (0.174 mL, 1.0 mmol) and stirring at RT for 19 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 8%] followed by purification using reverse phase column chromatography [Cl8 column, eluting with ACN in water (containing 0.1% HCl) from 0-50%] (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(pyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (272c) (56 mg, 57% yield) HCI as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.46 (s, 1H), 8.99 (s, 1H), 8.68 (s, 2H), 8.64 (s, 1H), 8.28 (d, J= 4.9 Hz, 1H), 7.99 (d, J= 8.4 Hz, 1H), 7.93 - 7.83 (m,

2H), 7.77 (t, J= 8.3 Hz, 1H), 7.15 - 7.05 (m, 1H), 5.85 (d, J= 17.4 Hz, 1H), 5.48 (d, J= 17.2

Hz, 1H), 4.51-4.41 (m, 1H), 3.95 - 3.87 (m, 1H), 2.39-2.19 (m, 2H), 2.00- 1.84 (m, 1H), 1.16-1.04 (m, 1H), 0.87 - 0.73 (m, 1H); MS (ES+): 496.15 (M+l); (ES-): 494.10 (M-l); Analysis calculated for C24H20F3N7O2.L8HCl.3H2O: C, 46.86; H, 4.56; Cl, 10.37; N, 15.94; Found: C, 46.80; H, 4.06; Cl, 10.11; N, 15.62.

Scheme 273

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (273c)

Step-1: Préparation of ethyl 2-(4-amino-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-95 yl)acetate (273a)

Compound 273a was prepared according to the procedure reported in step-1 of scheme-62, from ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (200 mg, 0.573 mmol) in dioxane (18 mL) using pyridin-3-ylboronic acid (106 mg, 0.859 mmol; CAS # 1692-25-7), bis(triphenylphosphine)palladium(II) chloride (80 mg, 0.115 mmol), a solution of césium carbonate (280 mg, 0.859 mmol) in water (2.2 mL) and heating at 100 °C for 13 h.

This gave after workup a mixture of ethyl 2-(4-amino-6-(pyridin-3-yl)-9H-pyrimido[4,5b]indol-9-yl)acetate (273a) MS (ES+): 348.15 (M+l) and 2-(4-amino-6-(pyridin-3-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (273b) MS (ES+): 320.10 (M+l) which was used as such for the next step.

Step-2: Préparation of 2-(4-amino-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (273b)

483

Compound 273b was prepared according to the procedure reported in step-4 of scheme-17, from the above mixture of ethyl 2-(4-amino-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9yl)acetate (273a) and 2-(4-amino-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (273b) (0.573 mmol) in THF (5 mL) and methanol (5 mL) using a solution of lithium hydroxide hydrate (147 mg, 3.44 mmol) in water (5 mL) and stirring at RT for 18 h. This gave after work up 2-(4-amino-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (273b) which was used as such for the next step; MS (ES+): 320.10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (273c)

Compound 273c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (273b) (65 mg, 0.204 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (84 mg, 0.265 mmol), HATU (155 mg, 0.407 mmol), DIPEA (0.142 mL, 0.814 mmol) and stirring at RT for 21 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 8%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (273c) (48 mg, 40% yield)

HCl sait as a white solid; ’H NMR (300 MHz, DMSO-cfe) δ 10.78 (s, 1H), 9.45 (d, J= 2.2 Hz, 1H), 9.18 (d,J= 1.7 Hz, 1H), 9.14 (bs, 3H), 9.03 - 8.96 (m, 1H), 8.86 (dd,J=5.5, 1.3 Hz, 1H), 8.71 (s, 1H), 8.16-8.06 (m, 2H), 8.00 (d,J=8.2 Hz, 1H), 7.91 (d, J= 8.7 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.85 (d, J= 17.4 Hz, 1H), 5.48 (d, J= 17.2 Hz, 1H), 4.43 (dd, J= 9.0, 5.5 Hz, 1H), 4.01 - 3.87 (m, 1H), 2.43 - 2.14 (m, 2H), 2.01 - 1.83 (m, 1H), 1.18 - 1.01 (m, 1H), 0.90 - 0.71 (m, 1H); MS (ES+): 583.10 & 585.10 (M+l), MS (ES-): 581.10 & 583.10 (M-l); Analysis calculated for C₂₈H₂₃BrN₈O₂.2.25HC1.4.65 H₂O: C, 44.89; H, 4.65; N, 14.96; Cl, 10.65; Found: C, 45.29; H, 4.52; N, 14.35.; Cl, 10.37.

Scheme 274

Ί

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (274g)

Step-1 : Préparation of 2,2,2-trifluoro-N-(2-iodo-4-(trifluoromethoxy)phenyl)acetamide (274b)

Compound 274b was prepared according to the procedure reported in step-1 of scheme-46, from 2-iodo-4-(trifluoromethoxy)aniline (274a) (10 g, 33.0 mmol; CAS # 845866-79-7) in DCM (50 mL) using triethylamine (11.50 mL, 83 mmol), trifluoroacetic acid anhydride (6.88 mL, 49.5 mmol) and stirring at RT for 15 h. This gave after workup 2,2,2-trifluoro-N-(210 iodo-4-(trifluoromethoxy)phenyl)acetamide (274b) (15.94 g) which was used as such for next step; MS (ES-): 397.90 (M-l).

Step-2: Préparation of 2-amino-5-(trifluoromethoxy)-lH-indole-3-carbonitrile (274c)

Compound 274c was prepared according to the procedure reported in step-1 of scheme-11, from 2,2,2-trifluoro-N-(2-iodo-4-(trifluoromethoxy)phenyl)acetamide (274b) (13.17 g, 33 mmol) in DMSO (40 mL) using malononitrile (2.62 g, 39.6 mmol), L-proline (0.760 g, 6.60 mmol), Cul (0.628 g, 3.30 mmol), a solution of K2CO3 (9.12 g, 66.0 mmol) in water (40 mL) and heating at 60 °C for 19 h under .an argon atmosphère. This gave after workup and purification using flash column chromatography [S1O2 gel (120 g), eluting with EtOAc in hexane from 0-66%] 2-amino-5-(trifluoromethoxy)-lH-indole-3-carbonitriIe (274c) (4.7 g,

59% yield) as a yellow solid; ’HNMR (300 MHz, DMSO-îZ₆) δ 10.93 (s, 1H), 7.17 (d, J= 8.5

Hz, 1H), 7.03 - 6.98 (m, 3H), 6.88 - 6.82 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -56.92; MS (ES+): 242.00 (M+l); MS (ES-): 240.00 (M-l).

485

Step-3: Préparation of 6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-4-amine (274d)

Compound 274d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5-(trifluoromethoxy)-lH-indole-3-carbonitrile (274c) (4.5 g, 18.66 mmol) in éthanol (85 mL) using formamidine acetate (15.70 g, 149 mmol) and refluxing for 19 h. This 5 gave after work up 6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-4-amine (274d) (2.873 g) as a gray solid which was used as such for the next step; 'H NMR (300 MHz, DMSO-î/ô) δ 12.06 (s, 1H), 8.42 (d, J= 2.3 Hz, 1H), 8.27 (s, 1H), 7.50 (d, J= 8.7 Hz, 1H), 7.39 - 7.29 (m, 3H); ¹⁹F NMR (282 MHz, DMSO-î/ô) δ -56.94; MS (ES+): 269.10 (M+l); MS (ES-): 267.00 (M-l).

Step-4: Préparation of ethyl 2-(4-amino-6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-9yl)acetate (274e)

Compound 274e was prepared according to the procedure reported in step-1 of scheme-1, from 6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-4-amine (274d) (1.2 g, 4.47 mmol) in DMF (25 mL) using ethyl 2-bromoacetate (0.546 mL, 4.92 mmol), CS2CO3 (3.64 g, 11.19 15 mmol) and stirring at RT for 20 h. This gave after work up ethyl 2-(4-amino-6(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-9-yl)acetate (274e) (495 mg, 31% yield) as an off-white solid; Ή NMR (300 MHz, DMSO-î/ô) δ 8.48 (d, J= 2.3 Hz, 1H), 8.32 (s, 1H), 7.72 (d, J= 8.9 Hz, 1H), 7.51 (s, 2H), 7.44 - 7.37 (m, 1H), 5.27 (s, 2H), 4.14 (q, J= 7.1 Hz, 2H), 1.20 (t, J= 7.1 Hz, 3H); ¹⁹F NMR (282 MHz, DMSO-î/ô) δ -56.97; MS (ES+): 355.10 (M+l).

Step-5: Préparation of 2-(4-amino-6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (274f)

Compound 274f was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-9-yl)acetate (274e) (400 mg, 1.129 mmol) in THF (6 mL) and MeOH (6 mL) using a solution of lithium hydroxide hydrate (290 mg, 6.77 mmol) in water (6 mL) and stirring at RT for 17 h. This gave after work up 2-(4-amino-6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (274f) (356 mg, 97% yield); Ή NMR (300 MHz, DMSO-î/ô) δ 13.09 (s, 1H), 8.49 - 8.46 (m, 1H), 8.32 (s, 1H), 7.71 (d, J= 8.9 Hz, 1H), 7.49 (s, 2H), 7.43 - 7.37 (m, 1H), 5.17 (s, 2H); ^I9F NMR (282 MHz, DMSO-î/ô) δ -56.96; MS (ES+): 327.10 (M+l); MS (ES-): 325.10 (M30 1).

486

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethoxy)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (274g)

Compound 274g was prepared according to the procedure reported in step-3 of scheme-1, 5 from 2-(4-amino-6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (274f) (75 mg, 0.230 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (88 mg, 0.276 mmol), HATU (175 mg, 0.460 mmol), DIPEA (0.160 mL, 0.92 mmol) and stirring at RT for 23 h. This gave after workup and purification by flash column chromatography [silica gel (25 g), eluting with MeOH in

DCM from 0 - 5%] followed by purification using reverse phase column chromatography

[CI8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0100%](lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (274g) (87 mg, 64% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î7₆) δ 10.77 (s, 1H), 8.83 (s, 2H), 15 8.71 - 8.64 (m, 2H), 8.00 (d, J= 8.2 Hz, 1H), 7.82 (d, J= 9.0 Hz, 1H), 7.70 (t, J= 8.0 Hz,

1H), 7.62 - 7.54 (m, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.81 (d, J= 17.4 Hz, 1H), 5.45 (d, J= 17.3

Hz, 1H), 4.41 (dd, J= 9.1, 5.5 Hz, 1H), 3.99 - 3.84 (m, 1H), 2.41 - 2.14 (m, 2H), 2.03 -1.77 (m, 1H), 1.16 - 0.99 (m, 1H), 0.85 - 0.71 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-cfc) δ -57.02;

MS (ES+): 590.10 & 592.10 (M+l); MS (ES-): 588.00 & 590.00 (M-l); Analysis calculated 20 for C24H19BrF3N7O3.HCl.H2O: C, 44.70; H, 3.44; N, 15.21; Cl, 5.50; Found: C, 44.48; H, 3.23; N, 15.18.; Cl, 5.42.

Scheme 275

487

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(furan-3-yl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yi)-2-azabicyclo[3.1.0]hexane-3-carboxamide (275c)

Step-1: Préparation of ethyl 2-(4-amino-6-(furan-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate 5 (275a)

Compound 275a was prepared according to the procedure reported in step-1 of scheme-62, from ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (200 mg, 0.573 mmol) in dioxane (18 mL) using furan-3-ylboronic acid (96 mg, 0.859 mmol), bis(triphenylphosphine)palladium(II) chloride (80 mg, 0.115 mmol) a solution of césium carbonate (280 mg, 0.859 mmol) in water (2.2 mL) and heating at 100 °C for 19 h. This gave after workup a mixture of ethyl 2-(4-amino-6-(furan-3-yl)-9H-pyrimido[4,5-b]indol-9yl)acetate (275a) and 2-(4-amino-6-(furan-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (275b) which was used as such for the next step; MS (ES+): 337.10 (M+l) and MS (ES+): 309.10 (M+l).

Step-2: Préparation of 2-(4-amino-6-(furan-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (275b)

Compound 275b was prepared according to the procedure reported in step-4 of scheme-17, from the above mixture of ethyl 2-(4-amino-6-(furan-3-yl)-9H-pyrimido[4,5-b]indol-9

488 yl)acetate (275a) and 2-(4-amino-6-(furan-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (275b) (0.573 mmol) in THF (5 mL) and methanol (5 mL) using a solution of lithium hydroxide hydrate (147 mg, 3.44 mmol) in water (5 mL) and stirring at RT for 18 h. This gave after work up 2-(4-amino-6-(furan-3-yl)-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (275b) (166 mg, 94% yield) which was used as such for the next step; MS (ES+): 309.10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(fiiran-3-yl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (275c)

Compound 275c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(furan-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (275b) (78 mg, 0.253 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (97 mg, 0.304 mmol), HATU (192 mg, 0.506 mmol), DIPEA (0.176 mL, 1.012 mmol) and stirring at RT for 13 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] ( 1 R,3 S,5R)-2-(2-(4-amino-6-(furan-3 -yl)-9H-pyrimido [4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (275c) (10 mg, 7% yield) HCl sait as a light yellow solid; *H NMR (300 MHz, DMSO-J₆) δ 10.77 (s, 1H), 8.69 (s, 1H), 8.64 - 8.48 (m, 3H), 8.26 (s, 1H), 8.01 (d, J= 8.1 Hz, 1H), 7.86 - 7.81 (m, 1H), 7.79 (t, J= 1.7 Hz, 1H), 7.74 - 7.66 (m, 2H), 7.34 - 7.29 (m, 1H), 7.18 - 7.16 (m, 1H), 5.75 (d, J= 17.3 Hz, 1H), 5.42 (d, J= 17.3 Hz, 1H), 4.42 (dd, J= 9.0, 5.6 Hz, 1H), 3.98 - 3.81 (m, 1H), 2.42 - 2.13 (m, 2H), 2.00 - 1.83 (m, 1H), 1.14 -1.01 (m, 1H), 0.83 - 0.69 (m, 1H); MS (ES+): 572.10 & 574.10 (M+l); MS (ES-): 570.10 & 572.10 (M-l).

Scheme 276

Préparation of (lR,3S,5R)-2-(2-(4-amino-6,7-dimethoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (276g)

Step-1: Préparation ofN-(2-bromo-4,5-dimethoxyphenyl)-2,2,2-trifluoroacetamide (276b)

Compound 276b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-4,5-dimethoxyaniline (276a) (5 g, 21.54 mmol; CAS # 16791-41-6) in DCM (75 mL) using triethylamine (5.10 mL, 36.6 mmol), trifhioroacetic acid anhydride (4.49 mL, 32.3 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (40 g), eluting with EtOAc in hexane from 0-50%] N-(210 bromo-4,5-dimethoxyphenyl)-2,2,2-trifluoroacetamide (276b) (1.49 g, 21% yield) as a paleyellow solid which was used as such for next step; *H NMR (300 MHz, DMSO-î/ô) δ 11.13 (s, 1H), 7.26 (s, 1H), 7.05 (s, 1H), 3.81 (s, 3H), 3.76 (s, 3H); MS (ES+): 327.90 (M+l). Step-2: Préparation of 2-amino-5,6-dimethoxy-lH-indole-3-carbonitrile (276c)

Compound 276c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-4,5-dimethoxyphenyl)-2,2,2-trifluoroacetamide (276b) (1.45 g, 4.42 mmol) in DMSO (5 mL) using malononitrile (0.334 mL, 5.30 mmol), L-proline (0.102 g, 0.884 mmol), Cul (84 mg, 0.442 mmol), a solution of K2CO3 (1.222 g, 8.84 mmol) in water (5 mL)

490 and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [silica gel (40 g), eluting with EtOAc in hexane from 0-50%] 2-amino-5,6dimethoxy-lH-indole-3-carbonitrile (276c) (457 mg, 48% yield) as a yellow solid; 'HNMR (300 MHz, DMSO-î/ô) δ 10.38 (s, 1H), 6.80 (s, 1H), 6.71 (s, 1H), 6.48 (s, 2H), 3.74 (s, 3H), 5 3.71 (s, 3H); MS (ES+): 218.10 (M+l).

Step-3: Préparation of 6,7-dimethoxy-9H-pyrimido[4,5-b]indol-4-amine (276d)

Compound 276d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5,6-dimethoxy-lH-indole-3-carbonitrile (276c) (0.45 g, 2.072 mmol) in éthanol (5 mL) using trimethyl orthoformate (2.266 mL, 20.72 mmol), NH4OAC (0.479 g,

6.21 mmol) and heating at 90 °C for 16 h. This gave after work up and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] 6,7-dimethoxy-9H-pyrimido[4,5-b]indoI-4-amine (276d) (282 mg, 49% yield) HCl sait as a pale-yellow solid; ’H NMR (300 MHz, DMSO-ôfc) δ 12.87 (s, 1H), 8.70 - 8.55 (m, 2H), 8.49 (s, 1H), 8.03 (s, 1H), 7.11 (s, 1H), 3.89 (s, 3H),

3.88 (s, 3H); MS (ES+): 245.10 (M+l).

Step-4: Préparation of ZerZ-butyl 2-(4-amino-6,7-dimethoxy-9H-pyrimido[4,5-b]indol-9yl)acetate (276e)

Compound 276e was prepared according to the procedure reported in step-1 of scheme-1, from HCl sait of 6,7-dimethoxy-9H-pyrimido[4,5-b]indol-4-amine (276d) (275 mg, 0.980 mmol) in DMF (7 mL) using ZerZ-butyl 2-bromoacetate (0.159 mL, 1.078 mmol), CS2CO3 (702 mg, 2.155 mmol) and stirring at RT for 16 h. This gave after work up and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] ZerZ-butyl 2-(4-amino-6,7-dimethoxy-9Hpyrimido[4,5-b]indol-9-yl)acetate (276e) (283 mg, 81% yield) as a pale yellow solid; 'H

NMR (300 MHz, DMSO-cfc) δ 8.63 (s, 2H), 8.54 (s, 1H), 8.06 (s, 1H), ΊΑΊ (s, 1H), 5.28 (s, 2H), 3.90 (s, 3H), 3.88 (s, 3H), 1.42 (s, 9H); MS (ES+): 359.2 (M+l).

Step-5: Préparation of 2-(4-amino-6,7-dimethoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (276f)

Compound 276f was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6,7-dimethoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (276e) (270 mg, 0.753 mmol) using 20% TFA in DCM (4324 pL, 11.30 mmol) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-6,7-dimethoxy-9H-pyrimido[4,5-b]indol

491

9-yl)acetic acid (276f) (311 mg, 99% yield) as a pale-yellow solid; ’H NMR (300 MHz, DMSO-Je) δ 8.50 (s, 1H), 8.47 (s, 2H), 8.03 (s, 1H), 7.51 (s, 1H), 5.27 (s, 2H), 3.89 (s, 3H), 3.89 (s, 3H); MS (ES+): 303.1 (M+l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6,7-dimethoxy-9H-pyrimido[4,5-b]indol-95 yI)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (276g)

Compound 276g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6,7-dimethoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (276f) (65.3 mg, 0.157 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (50mg, 0.157 mmol), HATU (90 mg,

0.235 mmol), DIPEA (0.137 mL, 0.785 mmol) and stirring at RT for 16 h. This gave after work up and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] ( 1 R,3S,5R)-2-(2-(4-amino-6,7-dimethoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N15 (6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (276g) (68 mg, 76% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/ô) δ 10.73 (s, 1H, D₂O exchangeable), 8.69 (s, 2H, D₂O exchangeable), 8.55 (s, 1H), 8.06 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.72 (t, J = 8.0 Hz, 1H), 7.36 - 7.29 (m, 2H), 5.72 (d, J= 17.3 Hz, 1H), 5.46 (d, J= 17.2 Hz, 1H), 4.42 (dd, J= 9.1, 5.7 Hz, 1H), 3.93 - 3.89 (m, 7H), 2.35 (dd, J= 13.4, 9.2 Hz, 1H), 2.29 - 2.14 (m, 1H), 1.98 - 1.85 (m, 1H), 1.08 (dt, J= 8.3, 5.4 Hz, 1H), 0.75 (td, J= 5.2, 2.4 Hz, 1H).

MS (ES+): 566.1/568.1 (M+l); (ES-): 564.1/566.1 (M-l); Analysis calculated for C₂5H₂4BrN₇O4.2.5H2O.1.25HCl.:C, 45.70; H, 4.64; Cl, 6.75; N, 14.92; Found: C, 45.82; H, 4.63; Cl, 6.58; N, 14.82.

Scheme 277

276f

277a

492

Préparation of (lR,3S,5R)-2-(2-(4-amino-6,7-dimethoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1,0]hexane-3-carboxamide (277a)

Compound 277a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6,7-dimethoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (276f) (65.1 mg, 0.156 mmol) in DMF (1.5 mL) using HCI sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (52 mg, 0.156 mmol), HATU (89 mg, 0.234 mmol), DIPEA (0.136 mL, 0.782 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with

DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCI) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6,7-dimethoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (277a) (51 mg, 56% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.72 (s, 1H, D₂O exchangeable), 8.70 (s, 2H, D₂O exchangeable), 8.55 (s, 1H), 8.10 - 7.91 (m, 2H), 7.71 (t, J= 8.0 Hz, 1H), 7.38 - 7.29 (m, 2H), 5.68 (d, J= 17.3 Hz, 1H), 5.40 (d, J= 17.2 Hz, 1H), 4.37 (dd, J= 9.1, 6.1 Hz, 1H), 3.89 (s, 6H), 3.69 (dd, J= 5.6, 2.4 Hz, 1H), 2.59 - 2.54 (m, 1H), 2.00 (dd, J= 13.3, 6.0 Hz, 1H), 1.32 (s, 3H), 1.08-0.96 (m, 1H), 0.96-0.82 (m, 1H). MS (ES+): 580.1/582.1 (M+l); (ES-): 578.1/580.1 (M-l); Analysis calculated for C₂6H₂₆BrN₇O₄

2.75H₂O. HCl.: C, 46.86; H, 4.92; Cl, 5.32; N, 14.71; Found: C, 46.80; H, 4.88; Cl, 5.17; N,

14.55.

Scheme 278

493

278f

278g

Préparation of (lR,3S,5R)-2-(2-(4-amino-6,8-dimethoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (278g)

Step-1: Préparation ofN-(2-bromo-4,6-dimethoxyphenyl)-2,2,2-trifluoroacetamide (278b)

Compound 278b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-4,6-dimethoxyaniline (278a) (1 g, 4.31 mmol; CAS # 197803-53-5) in DCM (15 mL) using triethylamine (1.021 mL, 7.33 mmol), trifluoroacetic acid anhydride (0.898 mL, 6.46 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (24 g), eluting with EtOAc in hexane from 0-50%] N-(210 bromo-4,6-dimethoxyphenyl)-2,2,2-trifluoroacetamide (278b) (0.89 g, 63% yield) as a pale yellow solid; 'HNMR (300 MHz, DMSO-J₆) δ 10.81 (s, 1H), 6.89 (d, J= 2.5 Hz, 1H), 6.72 (d, J= 2.6 Hz, 1H), 3.82 (s, 3H), 3.79 (s, 3H); MS (ES+): 327.91 (M+l).

Step-2: Préparation of 2-amino-5,7-dimethoxy-lH-indole-3-carbonitrile (278c)

494

Compound 278c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-4,6-dimethoxyphenyl)-2,2,2-trifluoroacetamide (278b) (0.87 g, 2.65 mmol) in DMSO (3 mL) and using malononitrile (0.200 mL, 3.18 mmol), L-proline (0.061 g, 0.530 mmol), Cul (51 mg, 0.265 mmol), a solution of K2CO3 (0.733 g, 5.30 mmol) in water (3 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [silica gel (40 g), eluting with EtOAc in hexane from 0-50%] 2-amino-5,7dimethoxy-lH-indole-3-carbonitrile (278c) (279 mg, 48% yield) as a yellow solid; *H NMR (300 MHz, DMSO-de) δ 10.63 (s, 1H), 6.31 (d, J= 2.1 Hz, 1H), 6.29 (s, 2H), 6.21 (d, J= 2.1 Hz, 1H), 3.83 (s, 3H), 3.73 (s, 3H); MS (ES+): 218.1 (M+l); (ES-): 216.0 (M-l).

Step-3: Préparation of 6,8-dimethoxy-9H-pyrimido[4,5-b]indol-4-amine (278d)

Compound 278d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5,7-dimethoxy-lH-indole-3-carbonitrile (278c) (0.270 g, 1.243 mmol) in éthanol (20 mL) using formamidine acetate (1046 mg, 9.94 mmol) and refluxing for 22 h. This gave after work up 6,8-dimethoxy-9H-pyrimido[4,5-b]indol-4-amine (278d) (206 mg, 15 68% yield) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-t/₆) δ 11.72 (s, 1H), 8.18 (s,

1H), 7.43 (d, J= 2.0 Hz, 1H), 7.12 (s, 2H), 6.60 (d, J= 2.0 Hz, 1H), 3.92 (s, 3H), 3.85 (s, 3H); MS (ES+): 245.1 (M+l).

Step-4: Préparation of ZerZ-butyl 2-(4-amino-6,8-dimethoxy-9H-pyrimido[4,5-b]indol-9yl)acetate (278e)

Compound 278e was prepared according to the procedure reported in step-1 of scheme-1, from 6,8-dimethoxy-9H-pyrimido[4,5-b]indol-4-amine (278d) (200 mg, 0.819 mmol) in DMF (6 mL) using terributyl 2-bromoacetate (0.133 mL, 0.901 mmol), CS2CO3 (400 mg, 1.228 mmol) and stirring at RT for 16 h. This gave after work up and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] terf-butyl 2-(4-amino-6,8-dimethoxy-9H-pyrimido[4,5b]indol-9-yl)acetate (278e) (198 mg, 68% yield) as a pale yellow solid; ’HNMR (300 MHz, DMSO-î/₆) δ 8.32 (s, 1H), 7.67 (s, 2H), 7.51 (d, J= 2.0 Hz, 1H), 6.67 (d, J= 2.0 Hz, 1H), 5.15 (s, 2H), 3.87 (s, 6H), 1.42 (s, 9H); MS (ES+): 359.1 (M+l).

Step-5: Préparation of 2-(4-amino-6,8-dimethoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (278f)

Compound 278f was prepared according to the procedure reported in step-2 of scheme-1, from teri-butyl 2-(4-amino-6,8-dimethoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (278e) (175

495 mg, 0.488 mmol) using 20% TFA in DCM (2803 pL, 7.32 mmol) and stirring at RT for 16 h. This gave after work up and purification TFA sait of 2-(4-amino-6,8-dimethoxy-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (278f) (225 mg, 97% yield) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.51 (s, 1H), 8.44 (s, 2H), 7.59 (d, J= 2.0 Hz, 1H), 6.76 (d, J = 2.0 Hz, 1H), 5.27 (s, 2H), 3.90 (s, 3H), 3.89 (s, 3H); MS (ES+): 303.1(M+l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6,8-dimethoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (278g)

Compound 278g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6,8-dimethoxy-9H-pyrimido[4,5-b]indol-9-yI)acetic acid (278f) (74.3 mg, 0.157 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (50 mg, 0.157 mmol), HATU (90 mg, 0.235 mmol), DIPEA (0.137 mL, 0.785 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6,8-dimethoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (278g) (63 mg, 71% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-c/ô) δ 10.74 (s, 1H, D₂O exchangeable), 8.68 (s, 2H, D₂O exchangeable), 8.57 (s, 1H), 8.02 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz,

1H), 7.61 (d, J= 2.0 Hz, 1H), 7.32 (d, 7.7 Hz, 1H), 6.75 (d, J= 2.0 Hz, 1H), 5.75 (d, J=

17.0 Hz, 1H), 5.53 (d, J= 16.9 Hz, 1H), 4.43 (dd, J= 8.9, 5.6 Hz, 1H), 3.91 (s, 3H), 3.88 (s, 3H), 3.86-3.78 (m, 1H), 2.39-2.14 (m, 2H), 2.00-1.85 (m, 1H), 1.15 - 1.05 (m, 1H), 0.71 - 0.60 (m, 1H). MS (ES+): 566.1/568.1 (M+l); (ES-): 564.1/566.1 (M-l); Analysis calculated for C₂5H₂₄BrN₇O4.1.5H₂O.1.15HCl: C, 47.26; H, 4.47; Cl, 6.42; N, 15.43; Found:

C, 47.47; H, 4.81; Cl, 6.46; N, 15.10.

Scheme 279

496

278f ^279a

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6,8-dimethoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (279a)

Compound 279a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6,8-dimethoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (278f) (74.3 mg, 0.157 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (52.2 mg, 0.157 mmol), HATU (90 mg, 0.236 mmol), DIPEA (0.137 mL, 0.785 mmol) and stirring at RT for 16 h.

This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6,8-dimethoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (279a) (53 mg, 58% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î7₆) δ 10.74 (s, 1H, D2O exchangeable), 8.68 (s, 2H, D2O exchangeable), 8.56 (s, 1H), 8.02 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.60 (s, 1H), 7.32 (d, J= 7.7 Hz, 1H), 6.75 (s, 1H), 5.70 (d, J = 17.0 Hz, 1H), 5.48 (d, J= 17.0 Hz, 1H), 4.38 (dd, J= 8.9, 5.9 Hz, 1H), 3.90 (s, 3H), 3.88 (s, 3H), 3.63-3.58 (m, 1H), 2.48-2.36 (m, 1H), 1.99 (dd, J= 13.4, 5.8 Hz, 1H), 1.31 (s, 3H),

1.08- 1.01 (m, 1H), 0.88 - 0.78 (m, 1H). MS (ES+): 580.2/582.1 (M+l); (ES-): 578.1/580.1 (M-l); Analysis calculated for C26H26BrN₇O₄.2H₂O.l.lHCl: C, 47.56; H, 4.77; Cl, 5.94; N, 14.93; Found: C, 47.47; H, 4.81; Cl, 5.88; N, 14.86.

Scheme 280

497

280f

280g

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-fluoro-8-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (280g)

Step-1 : Préparation of N-(2-bromo-4-fluoro-6-methoxyphenyl)-2,2,2-trifluoroacetamide (280b)

Compound 280b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-4-fluoro-6-methoxyaniline (280a) (1 g, 4.54 mmol; CAS # 354574-32-6) in DCM (15 mL) using triethylamine (1.077 mL, 7.73 mmol), trifluoroacetic acid anhydride (0.948 mL, 6.82 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with EtOAc in hexane from 050%] N-(2-bromo-4-fluoro-6-methoxyphenyl)-2,2,2-trifluoroacetamide (280b) (1.25 g, 87% yield) as a pale yellow solid; Ή NMR (300 MHz, DMSO-î7₆) δ 11.01 (s, 1H), 7.30 (dd, J= 8.2, 2.7 Hz, 1H), 7.18 (dd, J= 10.8, 2.7 Hz, 1H), 3.83 (s, 3H); MS (ES+): 315.90 (M+l). Step-2: Préparation of 2-amino-5-fhioro-7-methoxy-lH-indole-3-carbonitrile (280c)

Compound 280c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-4-fluoro-6-methoxyphenyl)-2,2,2-trifluoroacetamide (280b) (1.22 g, 3.86

498 mmol) in DMSO (5 mL) using malononitrile (0.292 mL, 4.63 mmol), L-proline (0.089 g, 0.772 mmol), Cul (74 mg, 0.386 mmol), a solution of K2CO3 (1.067 g, 7.72 mmol) in water (5 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [silica gel (40 g), eluting with EtOAc in hexane from 0-50%] 2-amino-5-fluoro5 7-methoxy-lH-indole-3-carbonitrile (280c) (538 mg, 68% yield) as a yellow solid; ’H NMR (300 MHz, DMSO-dô) δ 10.90 (s, 1H), 6.60 - 6.40 (m, 4H), 3.87 (s, 3H); MS (ES+): 206.10 (M+l); (ES-): 204.10 (M-l).

Step-3: Préparation of 6-fluoro-8-methoxy-9H-pyrimido[4,5-b]indol-4-amine (280d)

Compound 280d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5-fluoro-7-methoxy-lH-indole-3-carbonitrile (280c) (0.520 g, 2.53 mmol) in éthanol (30 mL) using formamidine acetate (2132 mg, 20.27 mmol) and refluxing for 22 h. This gave after work up AcOH sait of 6-fIuoro-8-methoxy-9H-pyrimido[4,5-b]indol-4-amine (280d) (665 mg, 90% yield) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-î7₆) δ 10.00 (s, 1H), 8.22 (s, 1H), 7.81 (dd, J= 9.Ί, 2.2 Hz, 1H), 7.18 (s, 2H), 6.92 (dd, J= 11.5, 2.2 Hz,

1H), 3.96 (s, 3H); MS (ES+): 233.1 (M+l).

Step-4: Préparation of ZerZ-butyl 2-(4-amino-6-fluoro-8-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetate (280e)

Compound 280e was prepared according to the procedure reported in step-1 of scheme-1, from AcOH sait of 6-fluoro-8-methoxy-9H-pyrimido[4,5-b]indol-4-amine (280d) (650 mg,

1.576 mmol) in DMF (15 mL) using ie/7-butyl 2-bromoacetate (0.256 mL, 0.1.734 mmol),

Cs2CO₃ (1284 mg, 3.94 mmol) and stirring at RT for 16 h. This gave after work up and purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] teri-butyl 2-(4-amino-6-fluoro-8methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (280e) (546 mg, 25% yield) as a pale yellow solid; ’H NMR (300 MHz, DMSO-îZ₆) δ 8.54 (s, 1H), 8.43 (s, 2H), 8.02 (dd, J= 9.4, 2.2 Hz, 1H), 7.11 (dd, J= 11.4, 2.2 Hz, 1H), 5.25 (s, 2H), 3.94 (s, 3H), 1.43 (s, 9H); MS (ES+): 347.1 (M+l).

Step-5: Préparation of 2-(4-amino-6-fluoro-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (280f)

Compound 280f was prepared according to the procedure reported in step-2 of scheme-1, from Zeri-butyl 2-(4-amino-6-fluoro-8-methoxy-9H-pyrimido[4,5-b]indoI-9-yl)acetate (280e) (125 mg, 0.361 mmol) using 20% TFA in DCM (2071 pL, 5.41 mmol) and stirring at RT for

499

h. This gave after work up TFA sait of 2-(4-amino-6-fluoro-8-methoxy-9H-pyrimido[4,5b]indol-9-yl)acetic acid (280f) (171 mg, 97% yield) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-îZô) δ 8.55 (s, 1H), 8.46 (s, 2H), 8.02 (dd, J= 9.4, 2.2 Hz, 1H), 7.13 (dd, J= 11.5, 2.2 Hz, 1H), 5.30 (s, 2H), 3.94 (s, 3H); MS (ES+): 291.10 (M+l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-fluoro-8-methoxy-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (280g)

Compound 280g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-fluoro-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (2801) (77 mg, 0.157 mmol) in DMF (7 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (50 mg, 0.157 mmol), HATU (90 mg, 0.235 mmol), DIPEA (0.137 mL, 0.785 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), DMA80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-fluoro-8-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (280g) (58 mg, 67% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-dô) δ 10.75 (s, 1H, D₂O exchangeable), 8.71 (s, 2H, D₂O exchangeable), 8.62 (s, 1H), 8.08 - 7.91 (m, 2H), 7.71 (t, J= 8.0 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 7.11 (dd, J= 11.5, 2.2 Hz, 1H), 5.78 (d, J= 17.1 Hz, 1H), 5.54 (d, J= 16.9 Hz, 1H), 4.43 (dd, J= 8.9, 5.6 Hz, 1H), 3.95 (s, 3H), 3.84 (ddd, J= 7.6, 5.3, 2.3 Hz, 1H), 2.41-2.14 (m, 2H), 2.00- 1.86 (m, 1H), 1.11 (dt, J= 9.8, 5.4 Hz, 1H), 0.66 (td, J =5.1,2.4 Hz, 1H). ¹⁹F NMR (282 MHz, DMSO) δ-117.06. MS (ES+): 554.1/556.1 (M+l); (ES-): 552.1/554.1 (M-l); Analysis calculated for C₂₄H2iBrFN₇O₃ 1.75H₂O. HCl.: C, 46.32; H, 4.13; Cl, 5.70; N, 15.75; Found: C, 46.24; H, 3.99; Cl, 5.70; N, 15.66.

Scheme 281

500

280f 281a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-fluoro-8-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (281a)

Compound 281a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-fluoro-8-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (280f) (77 mg, 0.157 mmol) in DMF (1.5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (52.2 mg, 0.157 mmol), HATU (90 mg, 0.236 mmol), DIPEA (0.137 mL, 0.785 mmol) and stirring at RT for

16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with DMA-80 in DCM from 0 -100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-fluoro-8-methoxy-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-375 carboxamide (281a) (65 mg, 73% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-dô) δ 10.74 (s, 1H, D2O exchangeable), 8.65 - 8.48 (m, 3H, 2H D2O exchangeable), 8.06 - 7.95 (m, 2H), 7.71 (t, J= 8.0 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 7.10 (dd, J= 11.5, 2.2 Hz, 1H), 5.73 (d, J= 17.0 Hz, 1H), 5.50 (d, J= 16.9 Hz, 1H), 4.38 (dd, J= 9.0, 5.9 Hz, 1H), 3.94 (s, 3H), 3.62 (dd, J= 5.5, 2.3 Hz, 1H), 2.49 - 2.41 (m, 1H), 2.00 (dd, J= 13.2, 5.9 Hz,

1H), 1.31 (s, 3H), 1.08 - 1.00 (m, 1H), 0.89 - 0.77 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-t/₆) δ -117.27. MS (ES+): 568.1/570.1 (M+l); (ES-): 566.1/568.1 (M-l); Analysis calculated for C25H23BrFN7O3.L75H2O.HCl.: C, 47.18; H, 4.36; Cl, 5.57; N, 15.41; Found: C, 47.11; H, 4.33; Cl, 5.23; N, 15.33

Scheme 282

501

257g 282a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-hydroxy-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (282a)

Compound 282a was prepared according to the procedure reported in scheme-164, from (lR,3S,5R)-2-(2-(4-amino-6-methoxy-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (257g) (64 mg, 0.116 mmol) in DCM (5 mL) using a solution of 1.0 M BBr₃ in DCM (0.581 mL, 0.581 mmol) and stirring at RT for 16 h. This gave after work up and purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-hydroxy-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (282a) (29 mg, 47% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 10.89 and 10.82 (2s, 1H, D2O exchangeable), 9.39 (s, 1H, D2O exchangeable), 8.55 (s, 1H), 8.47 (s, 2H, D2O exchangeable), 8.00 and 7.95 (2d, J= 8.1 Hz, 1H), 7.71 (t, J=

7.9 Hz, 1H), 7.65 (d, J= 2.1 Hz, 1H), 7.36 - 7.26 (m, 1H), 6.81 (s, 1H), 5.84 (d, J= 17.9 Hz,

1H), 5.57 (d, J= 17.9 Hz, 1H), 4.41 (dd, J= 9.0, 5.7 Hz, 1H), 3.95 - 3.82 (m, 1H), 2.65 and 2.61 (2s, 3H), 2.40-2.27 (m, 1H), 2.26-2.11 (m, 1H), 1.98- 1.85 (m, 1H), 1.14-0.99 (m, 1H), 0.73 - 0.55 (m, 1H); MS (ES+): 536/538 (M+l); (ES-): 534/536 (M-l).

Scheme 283

502

283d

283e

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5 -methyl-2-azabicyclo [3.1.0]hexane-3 carboxamide (283g)

Step-1: Préparation of N-(2-bromo-4-methylpyridin-3-yl)-2,2,2-trifluoroacetamide (283b)

Compound 283b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-4-methyIpyridin-3-amine (283a) (5 g, 26.7 mmol; CAS # 126325-50-6) in DCM (30 mL) using triethylamine (4.60 g, 45.4 mmol), trifluoroacetic acid anhydride (8.42 g, 40.1 mmol) and stirring at RT for 1 h. This gave after workup N-(2-bromo-410 methylpyridin-3-yl)-2,2,2-trifluoroacetamide (283b) (6.85 g, 91% yield) as a plum solid; Ή NMR (300 MHz, DMSO-î7₆) δ 11.51 (s, 1H), 8.30 (d, J = 4.9 Hz, 1H), 7.48 (d, J = 4.9 Hz, 1H), 2.25 (s, 3H). ¹⁹F NMR (282 MHz, DMSO- d₆) δ -74.21; MS (ES+): 283/285, (ES-): 281/283 (M-l).

Step-2: Préparation of 2-amino-7-methyl-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (283c)

Compound 283c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-4-methylpyridin-3-yl)-2,2,2-trifluoroacetamide (283b) (6.85 g, 24.20 mmol) in DMSO (30 mL) and using malononitrile (1.919 g, 29.0 mmol), L-proline (0.557 g,

503

4.84 mmol), Cul (0.461 g, 2.420 mmol), a solution of K2CO3 (6.69 g, 48.4 mmol) in water (30 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification using flash column chromatography [silica gel (40 g), eluting with MeOH in DCM from 0-6%] 2-amino-7-methyI-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (283c) (0.39 g, 9% yield) as a yellow solid; *H NMR (300 MHz, DMSO-î/ô) δ 11.66 (s, 1H), 7.98 (d, J =

5.6 Hz, 1H), 7.48 (s, 2H), 6.93 (d, J = 5.6 Hz, 1H), 2.43 (s, 3H); MS (ES+): 173 (M+l), (ES): 171 (M-l).

Step-3: Préparation of 8-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (283d)

Compound 283d was prepared according to the procedure reported in step-2 of scheme-29, 10 from 2-amino-7-methyl-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (283c) (0.39 g, 2.265 mmol) in éthanol (5 mL) using NH4OAC (0.524 g, 6.79 mmol), HC(OMe)₃ (2.404 g, 22.65 mmol) and heating at 90 °C for 16 h. This gave after work up 8-methyl-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (283d) (196 mg, 43% yield) as a brown solid; *HNMR (300 MHz, DMSO-î/ô) δ 12.11 (s, 1H), 847 - 8.22 (m, 2H), 8.01 (s, 1H), 7.19 75 (d, J= 4.9 Hz, 1H), 6.65 (s, 1H), 2.56 (s, 3H); MS (ES+): 200 (M+l), (ES-): 198 (M-l).

Step-4: Préparation of terZ-butyl 2-(4-amino-8-methyl-9H-pyrido[2',3':4,5]pyrroIo[2,3d]pyrimidin-9-yI)acetate (283e)

Compound 283e was prepared according to the procedure reported in step-1 of scheme-1, from 8-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (283d) (190 mg, 0.954 mmol) in DMF (15 mL) using ZerAbutyl 2-bromoacetate (205 mg, 1.049 mmol), CS2CO3 (622 mg, 1.908 mmol) and stirring at RT for 16 h. This gave after work up /er/-butyl 2-(4-amino8-methyl-9H-pyrido[2',3^,:4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (283e) (236 mg, 79% yield) as a white solid; Ή NMR (300 MHz, DMSO-î/ô) δ 8.50 - 8.31 (m, 2H), 7.90 (s, 1H), 7.20 (d, J= 5.0 Hz, 1H), 6.78 (s, 1H), 5.33 (s, 2H), 2.64 (s, 3H), 1.43 (s, 9H); MS (ES+): 314 (M+l), (ES-): 312 (M-l).

Step-5: Préparation of 2-(4-amino-8-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (283f)

Compound 283f was prepared according to the procedure reported in step-2 of scheme-1, from /er/-butyl 2-(4-amino-8-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-930 yl)acetate (283e) (236 mg, 0.753 mmol) in DCM (5 mL) using TFA (859 mg, 7.53 mmol) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-8-methyl-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (283f) (356 mg) as a white solid; ’H

504

NMR (300 MHz, DMSO-<7₆) δ 8.75 (s, 1H), 8.58 (s, 1H), 8.53 (d, J= 5.0 Hz, 1H), 7.93 (s, 1H), 7.37 (d, J= 5.0 Hz, 1H), 5.43 (s, 2H), 2.72 (s, 3H); MS (ES+): 258 (M+l), (ES-): 256 (M-l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,35 d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (283g)

Compound 283g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (283f) (75 mg, 0.202 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N10 (6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (67.2 mg, 0.202 mmol), HATU (92 mg, 0.242 mmol), DIPEA (131 mg, 1.010 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing

0.1% HCI) from 0-100%](lR,3S,5R)-2-(2-(4-amino-8-methyl-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicycIo[3.1.0]hexane-3-carboxamide (283g) (71 mg, 66% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.84 (s, 1H, D₂O exchangeable), 8.79 (s, 1H, D₂O exchangeable), 8.61 (s, 1H), 8.53 (d, J= 5.1 Hz, 1H), 8.34 (s, 2H, D₂O exchangeable), 8.01 (d, J= 8.2 Hz, 1H), 7.72 (t, J= 8.0 Hz, 1H), 7.42 (d, J= 5.3 Hz, 1H), 7.32 (d, J= 7.7 Hz,

1H), 5.88 (d, J= 18.0 Hz, 1H), 5.62 (d, J= 17.9 Hz, 1H), 4.39 (dd, J= 9.1, 6.1 Hz, 1H), 3.77 -3.68 (m, 1H), 2.75 (s, 3H), 2.49-2.40 (m, 1H), 1.98 (dd, J= 13.3, 6.0 Hz, 1H), 1.31 (s, 3H), 1.03 (t, J= 5.5 Hz, 1H), 0.87 (dd, J= 5.5, 2.3 Hz, 1H); MS (ES+): 535/537 (M+l); (ES): 533/535 (M-l); Analysis calculated for C₂₄H₂₃BrN₈O₂.1.45HC1.2.25H₂O: C, 45.84; H,

4.64; Cl, 8.18; N, 17.82; Found: C, 45.82; H, 4.56; Cl, 8.20; N, 17.70.

Scheme 284

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (284a)

Compound 284a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-9H-pyrido[2',3^,:4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (283f) (75 mg, 0.202 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (64.4 mg, 0.202 mmol), HATU (92 mg, 0.242 mmol) DIPEA (131 mg, 1.010 mmol) and stirring at RT for 16

h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%](lR,3S,5R)-2-(2-(4-amino-8-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (284a) (86 mg, 82% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-tfc) δ 10.84 (s, 1H, D2O exchangeable), 8.91 (bs, 1H, D2O exchangeable), 8.65 (s, 1H), 8.60 (bs, 1H, D2O exchangeable), 8.55 (d, J = 5.1 Hz, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.71 (t, J = 7.9 Hz, 1H), 7.46 (d, J = 5.2 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.95 (d, J = 18.0 Hz, 1H), 5.68 (d, J = 17.9 Hz, 1H), 4.47 - 4.39 (m, 1H), 3.97 - 3.90 (m, 1H), 2.78 (s, 3H), 2.43 - 2.29 (m, 1H), 2.29 20 2.15 (m, 1H), 2.02 - 1.81 (m, 1H), 1.19 - 1.01 (m, 1H), 0.79 - 0.66 (m, 1H); MS (ES+):

521/523 (M+l); (ES-): 519/521 (M-l).

Scheme 285

506

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (285g)

Step-1: Préparation of N-(2-bromo-6-methylpyridin-3-yl)-2,2,2-trifluoroacetamide (285b)

Compound 285b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-6-methylpyridin-3-amine (285a) (5 g, 26.7 mmol; CAS # 126325-53-9) in DCM (30 mL) using triethylamine (4.60 g, 45.4 mmol), trifluoroacetic acid anhydride (8.42 g, 40.1 mmol) and stirring at RT for 1 h. This gave after workup N-(2-bromo-610 methylpyridin-3-yl)-2,2,2-trifluoroacetamide (285b) (7.33 g, 97% yield) as a pale-yellow solid which was used as such for the next step; ’H NMR (300 MHz, DMSO-îZô) δ 11.42 (s, 1H), 7.81 (d, J = 7.9 Hz, 1H), 7.41 (d, J = 7.9 Hz, 1H), 2.50 (s, 3H). ¹⁹F NMR (282 MHz, DMSO-Jô) δ -74.21; MS (ES+): 283/285, (ES-): 281/283 (M-l).

Step-2: Préparation of 2-amino-5-methyl-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (285c)

507

Compound 285c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-6-methylpyridin-3-yl)-2,2,2-trifluoroacetamide (285b) (7.33 g, 25.90 mmol) in DMSO (30 mL) using malononitrile (2.053 g, 31.1 mmol), L-proline (0.596 g, 5.18 mmol), Cul (0.493 g, 2.59 mmol), a solution of K2CO3 (7.16 g, 51.8 mmol) in water (30 mL) 5 and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup 2-amino-5methyl-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (285c) (2.27 g, 51% yield) as apale-pink solid; Ή NMR (300 MHz, DMSO-cfc) δ 10.71 (s, 1H), 7.25 (d, J= 7.9 Hz, 1H), 7.05 (s, 2H), 6.72 (d, J= 7.9 Hz, 1H), 2.41 (s, 3H); MS (ES+): 173 (M+l), (ES-): 171 (M-l).

Step-3: Préparation of 6-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (285d)

Compound 285d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5-methyl-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (285c) (1.79 g, 10.40 mmol) in éthanol (10 mL) using NH4OAC (2.404 g, 31.2 mmol), HC(OMe)3 (11.03 g, 104 mmol) and heating at 90 °C for 16 h. This gave after work up 6-methyl-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (285d) (1.04 g, 50% yield) as a beige solid;

Ή NMR (300 MHz, DMSO-d₆) δ 11.88 (s, 1H), 8.32 (s, 1H), 7.73 (d, J= 8.2 Hz, 2H), 7.23 (d, J= 8.3 Hz, 1H), 6.53 (s, 1H), 2.63 (s, 3H); MS (ES+): 200 (M+l), (ES-): 198 (M-l). Step-4: Préparation of tert-butyl 2-(4-amino-6-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetate (285e)

Compound 285e was prepared according to the procedure reported in step-1 of scheme-1, from 6-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (285d) (1.02 g, 0.5.12 mmol) in DMF (15 mL) using tert-butyl 2-bromoacetate (1.099 g, 5.63 mmol), CS2CO3 (3.34 g, 10.24 mmol) and stirring at RT for 16 h. This gave after work up tert-butyl 2-(4-amino-6methyl-9H-pyrido[2^,,3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (285e) (1.24 g, 77% yield) as a beige solid; ‘H NMR (300 MHz, DMSO-î/₆) δ 8.36 (s, 1H), 7.92 (d, J= 8.4 Hz, 1H), 7.92 (s, 1H), 7.29 (d, J= 8.3 Hz, 1H), 6.58 (s, 1H), 5.13 (s, 2H), 2.65 (s, 3H), 1.39 (s, 9H); MS (ES+): 314 (M+l).

Step-5: Préparation of 2-(4-amino-6-methyl-9H-pyrido[2^,,3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (285f)

Compound 285f was prepared according to the procedure reported in step-2 of scheme-1, 30 from tert-butyl 2-(4-amino-6-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetate (285e) (1.23 g, 3.93 mmol) using TFA (4.48 g, 39.3 mmol) in DCM (5 mL) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-6-methyl-9H21436

508 pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (285f) (2.28 g) as a white solid; *H

NMR (300 MHz, DMSO-rf₆) δ 8.87 (bs,lH), 8.59 (s, 1H), 8.27 (d, J = 8.6 Hz, 1H), 7.90 (bs, 1 H), 7.51 (d, J = 8.5 Hz, 1H), 5.28 (s, 2H), 2.73 (s, 3H). ¹⁹F NMR (282 MHz, DMSO- d₆) δ 74.81; MS (ES+): 258 (M+l); (ES-): 256 (M-l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1,0]hexane-3-carboxamide (285g)

Compound 285g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (285f) (75 mg, 0.202 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (64.4 mg, 0.202 mmol), HATU (92 mg, 0.242 mmol), DIPEA (131 mg, 1.010 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCI) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (285g) (69 mg, 66% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/g) δ 10.76 (s, 1H, D₂O exchangeable), 8.86 (bs, 2H, D2O exchangeable), 8.61 (s, 1H), 8.33 (d, J = 7.7 Hz, 1H), 8.00 (d, J = 8.1 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.62 (d, J = 8.5 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.81 (d, J = 17.3 Hz, 1H), 5.45 (d, J = 17.3 Hz, 1H), 4.45 - 4.36 (m, 1H), 3.90 - 3.83 (m, 1H), 2.81 (s, 3H), 2.39 - 2.26 (m, 1H), 2.26 - 2.11 (m, 1H), 1.98 - 1.84 (m, 1H), 1.13 - 1.01 (m, 1H), 0.85 - 0.72 (m, 1H); MS (ES+): 521/523 (M+l); (ES-): 519/521 (M-l).

Scheme 286

Me ^285f 286a

509

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (286a)

Compound 286a was prepared according to the procedure reported in step-3 of scheme-1, 5 from TFA sait of 2-(4-amino-6-methyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (285f) (75 mg, 0.202 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (67.2 mg, 0.202 mmol), HATU (92 mg, 0.242 mmol), DIPEA (131 mg, 1.010 mmol) and stirring atRT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%](lR,3S,5R)-2-(2-(4-amino-6-methyl-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyI-2azabicyclo[3.1.0]hexane-3-carboxamide (286a) (65 mg, 60% yield) HCl sait as a white solid;

*H NMR (300 MHz, DMSO-àfc) δ 10.77 (s, 1H, D₂O exchangeable), 8.88 (bs, 2H, D₂O exchangeable), 8.61 (s, 1H), 8.33 (d, J= 8.9, 2.6 Hz, 1H), 8.00 (d, J = 8.1 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.62 (d, J = 8.6 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.77 (d, J = 17.3 Hz, 1H), 5.41 (d, J = 17.3 Hz, 1H), 4.43 - 4.30 (m, 1H), 3.67 - 3.64 (m, 1H), 2.81 (s, 3H), 2.49 - 2.42 (m, 1H), 1.98 (dd, J= 13.3, 5.8 Hz, 1H), 1.30 (s, 3H), 1.06 - 0.98 (m, 1H), 0.98 - 0.89 (m,

1H); MS (ES+): 535/537 (M+l), (ES-): 533/535 (M-l); Analysis calculated for

C₂4H₂₃BrN8O₂.1.95HCL4.25H₂O: C, 42.20; H, 4.94; Cl, 10.12; N, 16.40; Found: C, 42.44; H, 4.61; Cl, 9.93; N, 16.00.

Scheme 287

287a

510

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3carboxamide (287a)

Compound 287a was prepared according to the procedure reported in step-3 of scheme-1, 5 from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (100 mg, 0.228 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-chloropyridin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (269b) (65.8 mg, 0.228 mmol), HATU (130 mg, 0.342 mmol), DIPEA (147 mg, 1.141 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography

[silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (287a) (58 mg, 46% yield) HCl sait as a white solid;

’H NMR (300 MHz, DMSO-J_Ô) δ 10.81 (s, 1H, D₂O exchangeable), 8.79 (s, 1H), 8.64 (s, 1H), 8.64 (bs, 2H, D₂O exchangeable), 7.98 (d, J = 8.2 Hz, 1H), 7.81 (t, J = 8.0 Hz, 1H), 7.60 (s, 1H), 7.18 (d, J = 7.7 Hz, 1 H), 5.92 (d, J =18.0 Hz, 1H), 5.65 (d, J =17.9 Hz, 1H), 4.38 (dd, J = 9.1, 6.1 Hz, 1H), 3.74 - 3.69 (m, 1H), 2.76 (s, 3H), 2.49 - 2.36 (m, 1H), 1.98 (dd, J = 13.3, 6.1 Hz, 1H), 1.31 (s, 3H), 1.03 (t, J = 5.5 Hz, 1H), 0.89 - 0.82 (m, 1H); ¹⁹F NMR (282

MHz, DMSO-J₆) δ -58.77; MS (ES+): 558/560 (M+l); (ES-): 556/558 (M-l); Analysis calculated for C₂6H₂₃ClF₃N₇O₂.0.75HC1.2.25H₂O: C, 49.90; H, 4.55; Cl, 9.91; N, 15.67; Found: C, 50.06; H, 4.28; Cl, 9.86; N, 15.34.

Scheme 288

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(pyridin-3-yl)-9H-pyrimido[4,5b]indoI-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (288c)

Step-1 : Préparation of teri-butyl 2-(4-amino-8-methyl-6-(pyridin-3-yl)-9H-pyrimido[4,5b]indol-9-yl)acetate (288a)

Compound 288a was prepared according to the procedure reported in step-1 of scheme-263, from tert-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (192e) (300 mg, 0.767 mmol) in w-BuOH (4 mL) using pyridin-3-ylboronic acid (114c) (141 mg,

1.150 mmol), a solution of 1.27 M aqueous K3PO4 (1.207 mL, 1.533 mmol), Pdz(dba)3 (70.2 mg, 0.077 mmol), XPhos (73.1 mg, 0.153 mmol) and heating at 100 °C for 16 h under nitrogen. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 7%] Zer/-butyl 2-(4-amino-8-methyl-6(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (288a) (260 mg, 87% yield) as a white solid; ’H NMR (300 MHz, DMSO-î/₆) δ 9.11 (d, J = 2.4 Hz, 1H), 8.66 - 8.49 (m, 2H), 8.32 (s, 1H), 8.25 (dt, J = 8.1, 2.0 Hz, 1H), 7.58 (s, 1H), 7.56 - 7.35 (m, 3H), 5.37 (s, 2H), 2.71 (s, 3H), 1.44 (s, 9H); MS (ES+): 390 (M+l); (ES-): 388 (M-l).

512

Step-2: Préparation of 2-(4-amino-8-methyl-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (288b)

Compound 288b was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-8-methyl-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (288a) (260 mg, 0.668 mmol) in DCM (5 mL) using TFA (761 mg, 6.68 mmol) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-8-methyl-6-(pyridin-3-yl)9H-pyrimido[4,5-b]indol-9-yl)acetic acid (288b) (436 mg) and used as such for the next step; Ή NMR (300 MHz, DMSOW) δ 9.26 (d, J = 2.2 Hz, 1H), 8.87 - 8.71 (m, 2H), 8.68 (dd, J = 8.1, 2.2 Hz, 1H), 8.64 - 8.42 (m, 3H), 7.88 (dd, J = 8.2, 5.2 Hz, 1H), 7.81 (s, 1H), 5.48 (s,

2H), 2.78 (s, 3H); ¹⁹F NMR (282 MHz, DMSOW) δ -74.60; MS (ES+): 334 (M+l); (ES-):

332 (M-l). <

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(pyridin-3-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (288c)

Compound 288c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (288b) (75 mg, 0.168 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (53.4 mg, 0.168 mmol), HATU (76 mg, 0.201 mmol), DIPEA (108 mg, 0.838 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(pyridin-3-yl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (288c) (50 mg, 50% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 10.84 (s, 1H, D₂O exchangeable), 9.45 (d, J = 2.2 Hz, 1H), 9.12 (bs, 2H, D₂O exchangeable), 8.98 (dd, J = 8.4, 1.7 Hz, 1H), 8.94 (d, J = 1.7 Hz, 1H), 8.83 (d, 1H), 8.67 (s, 1H), 8.05 (dd, J = 8.3, 5.5 Hz, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.86 (s, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1 H), 5.99 (d, J = 18.0 Hz, 1H), 5.67 (d, J = 17.9 Hz, 1H),4.43 (dd, J = 9.1, 5.8 Hz,

1H), 4.01 - 3.94 (m, 1H), 2.80 (s, 3H), 2.37 (dd, J = 13.5, 9.1 Hz, 1H), 2.29 - 2.08 (m, 1H),

2.01 - 1.87 (m, 1H), 1.16 - 1.05 (m, 1H), 0.78 - 0.59 (m, 1H); MS (ES+): 597/599 (M+l);

513

(ES-): 595/597 (M-l); Analysis calculated for C29H25BrN₈O2.2.15HC1.4.25H₂O: C, 46.29; H, 4.78; Cl, 10.13; N, 14.89; Found: C, 46.42; H, 4.44; Cl, 10.13; N, 14.82.

Scheme 289

289a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyI-6-(pyridin-3-yl)-9H-pyrimido[4,5b]indoI-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (289a)

Compound 289a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (288b) (75 mg, 0.168 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (55.8 mg, 0.168 mmol), HATU (76 mg, 0.201 mmol), DIPEA (108 mg, 0.838 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [C 18 column (100 g), eluting with ACN in water (containing 0.1 % HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(pyridin-3-yl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (289a) (36 mg, 35% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-Jé) δ 10.84 (s, 1H, D₂O exchangeable), 9.40 (s, 1H), 9.14 - 8.84 (m, 4H, in which 2H were D₂O exchangeable), 8.84 - 8.78 (m, 1H), 8.66 (s, 1H), 8.10 - 7.95 (m, 2H), 7.85 (s, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.93 (d, J = 18.0 Hz, 1H), 5.64 (d, J = 17.9 Hz, 1H), 4.39 (dd, J = 9.0, 6.2 Hz, 1H), 3.73 (dd, J = 5.4, 2.4 Hz, 1H), 2.78 (s, 3H), 2.49 2.44 (m, 1H), 1.99 (dd, J = 13.4, 6.1 Hz, 1H), 1.32 (s, 3H), 1.04 (t, J = 5.4 Hz, 1H), 0.92 0.82 (m, 1H); MS (ES+): 611/613 (M+l), (ES-): 609/611 (M-l).

514

Scheme 290

290d 290e

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide 5 (290g)

Step-1: Préparation of N-(2,6-dibromopyridin-3-yl)-2,2,2-trifluoroacetamide (290b)

Compound 290b was prepared according to the procedure reported in step-1 of scheme-46, from 2,6-dibromopyridin-3-amine (290a) (10 g, 39.7 mmol; CAS # 39856-57-0) in DCM (30 mL) using triethylamine (6.83g, 67.5 mmol), trifluoroacetic acid anhydride (12.51 g, 59.5 mmol) and stirring at RT for 1 h. This gave after workup N-(2,6-dibromopyridin-3-yl)-2,2,2trifluoroacetamide (290b) (13.78 g, 100% yield) as a pale-yellow solid and was used as such for next step; ’H NMR (300 MHz, DMSO-^ δ 11.68 (s, 1H), 8.61 (d, J = 2.4 Hz, 1H), 8.35 (d, J = 2.3 Hz, 1H); ^I9F NMR (282 MHz, DMSO-J₆) δ -74.16; MS (ES-): 345/347 (M-l).

Step-2: Préparation of 2-amino-5-bromo-lH-pynOlo[3,2-b]pyridine-3-carbonitrile (290c)

515

Compound 290c was prepared according to the procedure reported in step-1 of scheme-11, fromN-(2,6-dibromopyridin-3-yl)-2,2,2-trifluoroacetamide (290b) (13.78 g, 39.6 mmol) in DMSO (30 mL) using malononitrile (3.14 g, 47.5 mmol), L-proline (0.912 g, 7.92 mmol), Cul (0.754 g, 3.96 mmol), a solution of K2CO3 (10.95 g, 79 mmol) in water (30 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and 2-amino-5bromo-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (290c) (7.03 g, 75% yield) as a beige solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.95 (s, 1H), 8.36 - 7.90 (m, 1H), 7.55 (d, J= 1.9 Hz, 1H), 7.39 (s, 2H); MS (ES+): 237/239 (M+l); (ES-): 235/237 (M-l).

Step-3: Préparation of 6-bromo-9H-pyrido[2',3':4,5]pynOlo[2,3-d]pyrimidin-4-amine (290d)

Compound 290d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5-bromo-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (290c) (7.02 g, 29.6 mmol) in éthanol (20 mL) using NH4OAC (11.41 g, 148 mmol), formamidine acetate (9.25 g, 89 mmol), HC(OMe)3 (31.4 g, 296 mmol) and heating at 90 °C for 16 h. This gave after work up 6-bromo-9H-pyrido[2^,,3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (290d) (7.68 g, 98% yield) as a beige solid; Ή NMR (300 MHz, DMSO-î/ô) δ 12.20 (s, 1H), 8.56 (d, J = 2.0 Hz, 1H), 8.37 (s, 1H), 8.03 (d, J = 2.0 Hz, 1H), 7.87 (s, 1H), 6.75 (s, 1H); MS (ES+): 264/266 (M+l), (ES-): 262/264 (M-l).

Step-4: Préparation of tert-buty! 2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetate (290e)

Compound 290e was prepared according to the procedure reported in step-1 of scheme-1, from 6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (290d) (3.00 g, 11.36 mmol) in DMF (15 mL) using ZerAbutyl 2-bromoacetate (2.437 g, 12.50 mmol), CS2CO3 (7.40 g, 22.72 mmol) and stirring at RT for 16 h. This gave after work up tert-butyl 2-(4amino-6-bromo-9H-pyrido[2',3^,:4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (290e) (3.20 g, 75% yield) as a beige solid; Ή NMR (300 MHz, DMSO-ifc) δ 8.61 (d, J = 1.9 Hz, 1H), 8.47 (d, J = 1.9 Hz, 1H), 8.40 (s, 1H), 7.97 (bs, 1H), 6.86 (bs, 1H), 5.16 (s, 2H), 1.41 (s, 9H); MS (ES+): 378/380 (M+l).

Step-5: Préparation of 2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (290f)

Compound 290f was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate

516 (290e) (300 mg, 0.793 mmol) using TFA (904 mg, 7.93 mmol) in DCM (5 mL) and stirring at RT for 16 h. This gave after work up TFA sait of 2-(4-amino-6-bromo-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (290f) (346 mg) as a beige solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.66 (d, J = 1.9 Hz, 1H), 8.58 (d, J = 1.9 Hz, 1H), 8.56 - 8.29 (m, 2H), 7.27 (s, 1H), 5.22 (s, 2H); ¹⁹F NMR (282 MHz, DMSO-t/_e) δ -74.62; MS (ES+):

322/324 (M+l); (ES-): 320/322 (M-l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (290g)

Compound 290g was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (290f) (75 mg, 0.172 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (54.8 mg, 0.172 mmol), HATU (78 mg, 0.206 mmol), DIPEA (111 mg, 0.860 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1,0]hexane-3-carboxamide (290g) (73 mg, 72% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ 10.80 (s, 1H, D₂O exchangeable), 9.07 (bs, 2H, D2O exchangeable), 8.78 - 8.70 (m, 2H), 8.61 (d, J = 2.0 Hz, 1H), 8.02 (d, J = 8.1 Hz, 1H), 7.71 (t, J = 7.9 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.83 (d, J = 17.4 Hz, 1H), 5.43 (d, J = 17.3 Hz, 1H), 4.47 - 4.41 (m, 1H), 3.93 - 3.84 (m, 1H), 2.40 - 2.28 (m, 1H), 2.28 - 2.16 (m, 1H), 1.99 - 1.84 (m, 1H), 1.14 - 0.96 (m, 1H), 0.96 - 0.74 (m,

1H); MS (ES+): 585/587 (M+l), (ES-): 583/585 (M-l); Analysis calculated for

C22Hi8Br2N8O2.LlHCl.2H2O: C, 39.89; H, 3.52; Cl, 5.89; N, 16.92; Found: C, 39.97; H, 3.29; Cl, 5.89; N, 16.88.

Scheme 291

517

291a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (291a)

Compound 291a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (100 mg, 0.228 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-chloropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (267b) (62.6 mg, 0.228 mmol), HATU (130 mg, 0.342 mmol), DIPEA (147 mg, 1.141 mmol) and stirring at RT for

16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 2%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-315 carboxamide (291a) (30 mg, 24% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-i/d) δ 10.82 (s, 1H, D₂O exchangeable), 8.81 (s, 1H), 8.78 (bs, 2H, D₂O exchangeable), 8.66 (s, 1H), 7.98 (d, J = 8.2 Hz, 1H), 7.82 (t, J = 8.0 Hz, 1H), 7.62 (s, 1H), 7.19 (d, J = 7.7 Hz, 1H), 5.98 (d, J = 18.0 Hz, 1H), 5.70 (d, J = 18.0 Hz, 1H), 4.43 (dd, J = 9.1, 5.7 Hz, 1H), 3.98 - 3.93 (m, 1H), 2.79 (s, 3H), 2.42 - 2.30 (m, 1H), 2.30 - 2.10 (m, 1H),

2.01 - 1.84 (m, 1H), 1.20 - 1.02 (m, 1H), 0.78 - 0.58 (m, 1H); ¹⁹F NMR (282 MHz, DMSOdè) δ -58.79; MS (ES+): 544/546 (M+l); (ES-): 542/544 (M-l); Analysis calculated for C25H21CIF3N-7O2.HCI.3H2O: C, 47.33; H, 4.45; N, 15.45; Found: C, 47.40; H, 4.05; N, 15.36.

Scheme 292

I

518

Br

292a

290f

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (292a)

Compound 292a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (290f) (75 mg, 0.172 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (57.2 mg, 0.172 mmol), HATU (78 mg, 0.206 mmol), DIPEA (111 mg, 0.860 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-bromo-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-215 azabicyclo[3.1.0]hexane-3-carboxamide (292a) (89 mg, 86% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.72 (s, 1H, D₂O exchangeable), 8.70 (bs, 1H, D₂O exchangeable), 8.63 (d, J = 1.9 Hz, 1H), 8.56 (s, 1H), 8.46 (d, J = 2.0 Hz, 1H), 8.14 (bs, 1H, D₂O exchangeable), 7.95 (d, J = 8.2 Hz, 1H), 7.63 (t, J = 7.9 Hz, 1H), 7.25 (d, J = 7.7 Hz, 1H), 5.67 (d, J = 17.4 Hz, 1H), 5.29 (d, J = 17.3 Hz, 1H), 4.32 (dd, J = 9.1, 5.8 Hz, 1H), 3.58

- 3.55 (m, 1H), 2.42 - 2.32 (m, 1H), 1.91 (dd, J= 13.3, 5.8 Hz, 1H), 1.24 (s, 3H), 1.00 - 0.83 (m, 2H); MS (ES+): 599/601 (M+l), (ES-): 597/599 (M-l); Analysis calculated for C₂3H₂₀Br₂N₈O₂.HC1.2H₂O: C, 41.06; H, 3.75; Cl, 5.27; N, 16.66; Found: C, 41.06; H, 3.51; Cl, 5.21; N, 16.52.

Scheme 293

519

(-)-Diastereomer-B 293B

Préparation of (lS,3R,5S)-2-(ll-amino-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2c][l,3]oxazine-6-carbonyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (293A; (-)-diastereoisomer-A), (lS,3R,5S)-2-(ll-amino-6H5 benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2-c][l,3]oxazine-6-carbonyl)-N-(6-bromopyridin-2-yl)2-azabicyclo[3.1.0]hexane-3-carboxamide (293B; (-)-diastereoisomer-B)

Compound (233h) was prepared according to procedure reported in scheme 233 from 11amino-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2-c][l,3]oxazine-6-carboxylic acid (233g) (171 mg, 0.606 mmol) and (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane10 3-carboxamide, HCl (4a) (212 mg, 0.666 mmol) in DMF (5 mL) using HATU (346 mg,

0.909 mmol), DIPEA (0.528 mL, 3.03 mmol) and stirring at RT for 48h. This gave after workup and purification by flash column chromatography [silica (12 g), eluting with ethyl acetate/MeOH 9:1 in hexanes from 0-100%] to afford Compound (233h) as a mixture of diastereomers. The mixture was repurified by chromatography [silica (12 g), eluting with ethyl acetate/MeOH 9:1 in DCM from 0-100%] to afford pure individual diastereoisomer 293A and 293B. The following was the order and data for the compound that was eluted

1. (lS,3R,5S)-2-(ll-amino-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2-c][l,3]oxazine-6carbonyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (293A; (-)-diastereoisomer-A) as a free base; Ή NMR (300 MHz, MeOD-<7₄) δ 8.11 (s, 1H),

7.95 (d, J= 8.4 Hz, 1H), 7.68 (d, J= 7.9 Hz, 1H), 7.52 (t, J= 8.2 Hz, 1H), 7.37 (s,

1H), 7.27 (t, J= 7.9 Hz, 1H), 7.21 - 7.03 (m, 3H), 6.92 (s, 1H), 4.33-4.11 (m, 2H), 2.47-2.19 (m, 2H), 2.05 - 1.99 (m, 1H), 1.27-1.23 (m, 1H), 0.79-0.59 (m, 1H); MS (ES+): 546.10, 548.10 (M+l); (ES-): 543.00, 544.00 (M-l); Optical rotation [a]_D

520 .

= -11.43 (c = 0.14, MeOH); Compound (293A; (-)-diastereoisomer-A) was further purified by reverse-phase column chromatography [EZ-PREP, C-18 column, 50 g, eluting with 0.1% aqueous HCl in water and acetonitrile from 0-100%] to afford (lS,3R,5S)-2-(l l-amino-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2-c][l,3]oxazine-65 carbonyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1,0]hexane-3-carboxamide (293A;

(-)-diastereoisomer-A) (42mg, 13 % yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ 10.76 (s, 1H), 9.58 (s, 1H, D₂O exchangeable), 8.76 (s, 1H, D₂O exchangeable), 8.45 (s, 1H), 7.93 (d, J= 8.2 Hz, 1H), 7.82 - 7.56 (m, 3H), 7.39 (d, J = 6.8 Hz, 2H), 7.33 - 7.04 (m, 3H), 4.46 - 4.26 (m, 1H), 4.26 - 4.06 (m, 1H), 2.34 10 2.18 (m, 1H), 2.18-2.01 (m, 1H), 2.01 - 1.81 (m, 1H), 1.36- 1.08 (m, 1H), 0.630.32 (m, 1H); Ή NMR (300 MHz, DMSO-W)) δ 10.71 (d, J= 4.3 Hz, 1H), 8.41 (d, J= 3.9 Hz, 1H), 7.91 (d, J= 8.3 Hz, 1H), 7.74 (q, J= 6.1, 4.5 Hz, 2H), 7.65 (t, J= 7.9 Hz, 1H), 7.48 - 7.31 (m, 2H), 7.31 - 7.07 (m, 3H), 4.41 - 4.22 (m, 1H), 4.22 4.05 (m, 1H), 2.35-2.17 (m, 1H), 2.17-2.00 (m, 1H), 2.00-1.82 (m, 1H), 1.21 (q,

J= 6.3 Hz, 1H), 0.62 - 0.39 (m, 1H); MS (ES+): 546.1, 548.1 (M+l); (ES-): 544.1,

546.1 (M-l); LCMS, Rt = 1.96 min, (wavelength = 254 nM), 98.06%; Optical rotation [a]_D = +5.88 (c = 0.17, MeOH);

2. (lS,3R,5S)-2-(l l-amino-6H-benzo[e]pyrimido[5',4':4,5]pyrrolo[l,2-c][l,3]oxazine-6carbonyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (293B;

(-)-diastereoisomer-B); Ή NMR (300 MHz, MeODX) δ 8.08 (s, 1H), 7.86 (d, J=

8.3 Hz, 1H), 7.67 (d, J= 7.6 Hz, 1H), 7.51 (t, J= 8.0 Hz, 1H), 7.25 - 7.14 (m, 4H), 7.12-7.04 (m, 1H), 6.94 (s, 1H), 4.53-4.38 (m, 1H), 4.23-4.13 (m, 1H), 2.432.25 (m, 2H), 2.03 - 1.97 (m, 1H), 1.30-1.22 (m, 1H), 1.02-0.91 (m, 1H); MS (ES+): 546.10, 548.10 (M+l); Optical rotation [a]_D= -223.88 (c = 0.135, MeOH),

Compound (293B;

(-)-diastereoisomer-B) was further purified by reverse-phase column chromatography [EZ-PREP, C-18 column, 50 g, eluting with 0.1% aqueous HCl in water and acetonitrile from 0-100%] to afford (lS,3R,5S)-2-(ll-amino-6Hbenzo[e]pyrimido[5',4':4,5]pyrrolo[l,2-c][l,3]oxazine-6-carbonyl)-N-(630 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (293B; (-)diastereoisomer-B) to afford (64mg, 19% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-iZé) δ 10.79 (s, 1H), 9.60 (s, 1H, D₂O exchangeable), 8.84 (s, 1H, D₂O exchangeable), 8.49 (s, 1H), 7.92 (d, J= 8.1 Hz, 1H), 7.84 - 7.62 (m, 2H), 7.57 21436

521

7.38 (m, 3H), 7.38 - 7.16 (m, 3H), 5.21 - 4.35 (m, 1H), 4.35 - 4.10 (m, 1H), 2.44 2.14 (m, 2H), 2.02 (p, J=6.8Hz, 1H), 1.34- 1.05 (m, 1H), 0.90 - 0.67 (m, 1H); 'H NMR (300 MHz, DMSO-ri₆/D₂O) δ 11.66 - 10.37 (m, 1H), 8.58 - 8.15 (m, 1H), 8.03 - 7.74 (m, 2H), 7.69 (t, J= 7.9 Hz, 1H), 7.56 - 7.14 (m, 6H), 5.14 - 4.34 (m, 1H),

4.15 (d, J= 6.5 Hz, 1H), 2.44-2.28 (m, 1H), 2.28 - 2.13 (m, 1H), 2.13 - 1.88 (m,

1H), 1.37- 1.08 (m, 1H), 0.93-0.65 (m, 1H); MS (ES+): 546.1, 548.1 (M+l); (ES-): 544.1, 546.1 (M-l); LCMS, Rt = 1.98 min, (wavelength = 254 nM), 98.60%; Optical rotation [œ]d= -256.15 (c = 0.26, MeOH);

Scheme 294

Préparation of (2S,4R)-l-(2-(4-aminoquinazolin-6-yl)acetyl)-N-(3-chloro-2-fluorobenzyl)-4fluoropyrrolidine-2-carboxamide (294c)

Step-1: Préparation of 2-(4-aminoquinazolin-6-yl)acetic acid (294b)

Compound 294b was prepared according to the procedure reported in step-1 of scheme-2, from 6-bromoquinazolin-4-amine (294a) (1.00 g, 4.46 mmol; CAS # 21419-48-7) using zinc (1.752 g, 26.8 mmol), TMSC1 (0.170 mL, 1.339 mmol) in THF (5 mL) and Pd₂(dba)₃ (0.204 g, 0.223 mmol) and XPhos (0.213 g, 0.446 mmol) in THF (10 mL) and heating at 60 °C under argon for 16 h. The cooled reaction mixture was diluted with H₂O (30 mL) and EtOAc (30 mL), followed by addition of solid NH4CI (2 g). After 15-min stirring at rt, TFA (10 mL) was added and stirring continued for 30 min. The mixture was then filtered. The filtered cake was washed thoroughly with H₂O and EtOAc and filtrate was concentrated in vacuum to provide the product TFA sait of 2-(4-aminoquinazolin-6-yI)acetic acid (294b) (475 mg, 33.5 % yield) as a brown solid. Ή NMR (300 MHz, DMSO-cfc) δ 9.70 (s, 2H), 8.75 (s, 1H), 8.24 (d, J = 1.7 Hz, 1H), 7.90 (dd, J = 8.5, 1.7 Hz, 1H), 7.69 (d, J = 8.6 Hz, 1H), 3.72 (s, 2H). ^,9F

NMR (282 MHz, DMSO-cfc) δ -74.74; MS (ES+): 204 (M+l); (ES-): 202 (M-l).

522

Step-2: Préparation of (2S,4R)-l-(2-(4-aminoquinazolin-6-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (294c)

Compound 294c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-aminoquinazolin-6-yl)acetic acid (294b) (163 mg, 0.515 mmol) in

DMF (5 mL) using (2S,4R)-N-(3-chloro-2-fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (7a) (200 mg, 0.515 mmol), HATU (235 mg, 0.617 mmol), DIPEA (0.449 mL, 2.57 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography (SiCh, 12 g, eluting with 0-5% MeOH in DCM) followed by reverse phase column chromatography [C-18 column, 100 g, eluting with 0.1% aqueous HCl in H2O and

MeCN from 0-100%] (2S,4R)-l-(2-(4-aminoquinazolin-6-yl)acetyl)-N-(3-chloro-2fluorobenzyl)-4-fluoropyrrolidine-2-carboxamide (294c) (21 mg, 9 % yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/ô) (mixture of two rotamers) δ 9.70 (bs, 2H, D2O exchangeable), 8.82 (s, 1H), 9.18 and 8.74 (2t, J= 5.9 Hz, 1H), 8.36 and 8.30 (2s, 1H), 8.02 7.84 (m, 1H), 7.84 - 7.72 (m, 1H), 7.53 - 7.39 (m, 1H), 7.40 - 7.25 (m, 1H), 7.23 - 7.04 (m,

1H), 5.60 - 5.21 (m, 1H), 4.51 - 3.72 (m, 6H), 3.60 - 3.46 (m, 1H), 2.76 - 2.40 (m, 1H), 2.23

- 1.86 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-ifc) δ -121.25, -121.65, -176.17, -176.38; MS (ES+) 460/462 (M+l).

Scheme 295

295a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (295a)

Compound 295a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-925 yl)acetic acid (184f) (75 mg, 0.171 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N21436

523

(6-bromopyrazin-2-yl)-2-azabicycIo[3.1.0]hexane-3-carboxamide (112a) (54.7 mg, 0.171 mmol), HATU (98 mg, 0.257 mmol), DIPEA (111 mg, 0.856 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 3%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (295a) (66 mg, 65% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-e/d) δ 11.22 (s, 1H, D₂O exchangeable), 9.23 (s, 1H), 8.92 (bs, 2H, D₂O exchangeable), 8.82 (s, 1H), 8.70 (s, 1H), 8.53 (s, 1H), 7.64 (s, 1H), 6.00 (d, J = 18.0 Hz, 1H), 5.71 (d, J = 17.9 Hz, 1H), 4.47 (dd, J = 9.0, 5.7 Hz, 1H), 4.00 - 3.92 (m, 1H), 2.79 (s, 3H), 2.44 - 2.31 (m, 1H), 2.31 - 2.17 (m, 1H), 2.02 - 1.88 (m, 1H), 1.19 - 1.02 (m, 1H), 0.80 0.64 (m, 1H); ^I9F NMR (282 MHz, DMSO-J_Ô) δ -58.83; MS (ES+): 589/591 (M+l), (ES-): 587/589 (M-l).

Scheme 296

296a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (296a)

Compound 296a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (75 mg, 0.171 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (261c) (57.1 mg, 0.171 mmol), HATU (98 mg, 0.257 mmol), DIPEA (111 mg, 0.856 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica

524 gel (12 g), eluting with MeOH in DCM from 0 - 2%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-5-methyl-25 azabicyclo[3.1.0]hexane-3-carboxamide (296a) (63 mg, 61% yield) HCl sait as a white solid; 'HNMR(300 MHz, DMSO-ri₆) δ 11.21 (s, 1H, D₂O exchangeable), 9.24 (s, 1H), 8.91 (bs, 2H, D₂O exchangeable), 8.85 (s, 1H), 8.69 (s, 1H), 8.53 (s, 1H), 7.62 (s, 1H), 5.94 (d, J = 18.0 Hz, 1H), 5.66 (d, J = 18.0 Hz, 1H), 4.42 (dd, J = 9.1, 6.1 Hz, 1H), 3.76 - 3.71 (m, 1H), 2.76 (s, 3H), 2.57 - 2.51 (m, 1H), 2.10 - 1.94 (m, 1H), 1.31 (s, 3H), 1.08 - 0.98 (m, 1H), 0.94

- 0.82 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-ri_d) δ -58.84; MS (ES+): 603/605 (M+l), (ES-):

601/603 (M-l).

Scheme 297

297c

297d

Préparation of (1 R,3 S,5R)-2-(2-(4-amino-6-(3-methylureido)-9H-pyrimido[4,5-b] indol-915 yl)acetyI)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (297d)

Step-1: Préparation of l-(4-amino-9H-pyrimido[4,5-b]indol-6-yl)-3-methylurea (297a)

525

A suspension of 4-amino-9H-pyrimido[4,5-b]indole-6-carbonyl azide (225a) (111 mg, 0.438 mmol) in toluene (4 mL) was added methanamine hydrochloride (177 mg, 2.63 mmol), triethylamine (0.916 mL, 6.57 mmol) and heated at 115 °C for 2 h in a microwave. The reaction mixture was cooled to RT, triturated with dichloromethane/methanol (9:1) (30 mL), filtered, washed with dichloromethane/methanol (9:1). The filtrate was concentrated and purified by flash column chromatography [silica gel (40 g), eluting with dichloromethane/methanol (1:0 to 9:1)] to afford l-(4-amino-9H-pyrimido[4,5-b]indol-6-yl)3-methylurea (297a) (44 mg, 39% yield) as a light brown gum; MS (ES+): 257.10 (M+Na).

Step-2: Préparation of ZerZ-butyl 2-(4-amino-6-(3-methylureido)-9H-pyrimido[4,5-b]indol-9yl)acetate (297b)

Compound 297b was prepared according to the procedure reported in step-1 of scheme-1, from l-(4-amino-9H-pyrimido[4,5-b]indol-6-yl)-3-methylurea (297a) (42 mg, 0.164 mmol) in DMF (10 mL) using ZerZ-butyl 2-bromoacetate (0.027 mL, 0.180 mmol) and CS2CO3 (133 mg, 0.410 mmol) and stirring at RT for 17 h. This gave after work up ZerZ-butyl 2-(4-amino6-(3-methylureido)-9H-pyrimido[4,5-b]indol-9-yl)acetate (297b) (32 mg) and was used as such for the next step; MS (ES+): 371.20 (M+l).

Step-3: Préparation of 2-(4-amino-6-(3-methylureido)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (297c)

Compound 297c was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6-(3-methylureido)-9H-pyrimido[4,5-b]indol-9-yl)acetate (297b) (30 mg, 0.081 mmol) in DCM (10 mL) using TFA and stirring at RT. This gave after work which was used as such for the next step; MS (ES+): 315.10 (M+l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(3-methylureido)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (297d)

Compound 297d was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(3-methylureido)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (297c) (25.5 mg, 0.081 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (51.6 mg, 0.162 mmol), HATU (61.6 mg, 0.0.162 mmol), DIPEA (0.071 mL, 0.405 mmol) and stirring at RT for 21 h. This gave after workup and purification using flash column chromatography [silica gel (12

526

g), eluting with MeOH in DCM from 0-10%] followed by treating with acetonitrile (2 mL) and 0.1% aq. HCl (10 mL) to afford (lR,3S,5R)-2-(2-(4-amino-6-(3-methylureido)-9Hpyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (297d) (4 mg, 9% yield) HCl sait as a light brown solid; *H NMR (300 MHz, 5 DMSO-î/ô) δ 10.76 (s, 1H), 8.55 (s, 1H), 8.42 (s, 1H), 8.27 (s, 1H), 8.01 (d, J= 8.1 Hz, 1H),

7.70 (t, J= 7.9 Hz, 1H), 7.55 (s, 2H), 7.32 (d, J= 7.7 Hz, 1H), 6.19 (s, 1H), 5.71 (d, J= 17.5 Hz, 1H), 5.37 (d, J= 17.3 Hz, 1H), 4.51 -4.33 (m, 1H), 3.98-3.85 (m, 1H), 2.70-2.63 (m, 3H), 2.40-2.14 (m, 2H), 1.97 - 1.83 (m, 1H), 1.14 - 0.98 (m, 1H), 0.81 - 0.71 (m, 1H); MS (ES+): 578.20 (M+l); (ES-): 576.10 (M-l).

'0

Scheme 298

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-phenyl-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (298c)

Step-1: Préparation of ZerZ-butyl 2-(4-amino-6-phenyl-9H-pyrido[2',3':4,5]pynOlo[2,3d]pyrimidin-9-yl)acetate (298a)

527

Compound 298a was prepared according to the procedure reported in step-lof scheme-59, from tert-butyl 2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (290e) (300 mg, 0.793 mmol) in dioxane (4 mL) using phenyl boronic acid (145 mg, 1.190 mmol), bis(triphenylphosphine)palladium(II) chloride (55.7 mg, 0.079 mmol) a solution of

3.3 M potassium carbonate (0.721 mL, 2.380 mmol) and heating at 100 °C for 16 h. This gave after workup and purification using flash column chromatography [silica gel (24g), eluting with MeOH in DCM from 0-3%] tert-butyl 2-(4-amino-6-phenyl-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (298a) (281 mg, 94% yield) as a yellow solid; *HNMR (300 MHz, DMSO-î/ô) δ 8.85 (d, J= 1.8 Hz, 1H), 8.42 (d, J= 1.9 Hz, 1H),

8.39 (s, 1H), 8.15 - 7.66 (m, 3H), 7.61 - 7.50 (m, 2H), 7.49 - 7.40 (m, 1H), 6.85 (bs, 1H),

5.24 (s, 2H), 1.41 (s, 9H); MS (ES+): 376 (M+l).

Step-2: Préparation of 2-(4-amino-6-phenyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (298b) ' Compound 298b was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-6-phenyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (298a) (280 mg, 0.746 mmol) in DCM (5 mL) using TFA (850 mg, 7.46 mmol) and stirring at RT for 16 h. This gave after work up 2-(4-amino-6-phenyl-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetic acid (298b) (386 mg) TFA sait as a pale-yellow solid; ’H NMR (300 MHz, DMSO-î/ô) δ 8.98 (d, J= 1.8 Hz, 1H), 8.80 (bs, 1H), 8.65 (d, J= 1.9 Hz, 1H), 8.60 (s, 20 1H), 8.10 (bs, 1H), 7.92 - 7.86 (m, 2H), 7.65 - 7.52 (m, 2H), 7.53 - 7.42 (m, 1H), 5.36 (s,

2H); ¹⁹F NMR (282 MHz, DMSO) δ -74.76; MS (ES+): 320 (M+l); (ES-): 318 (M-l). Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-phenyl-9H-pyrido[2',3':4,5]pynOlo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1,0]hexane-3-carboxamide (298e)

Compound 298c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-phenyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (298b) (75 mg, 0.173 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (55.1 mg, 0.173 mmol), HATU (79 mg, 0.208 mmol), DIPEA (112 mg, 0.865 mmol) and stirring at RT for 16

h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [C18 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-phenyl-9H-pyrido[2',3':4,5]pyrrolo[2,3

528

d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (298c) (72 mg, 71% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-Jd) δ 10.79 (s, 1H, D₂O exchangeable), 9.15 - 8.85 (bs, 1H, D2O exchangeable), 8.98 (d, J = 1.8 Hz, 1H), 8.85 - 8.56 (bs, 1H, D₂O exchangeable), 8.70 (s, 1H,), 8.53 (d, J = 1.9 Hz, 1H), 7.98 (d, J = 5 8.2 Hz, 1H), 7.90 - 7.83 (m, 2H),, 7.67 (t, J = 8.0 Hz, 1H), 7.63 - 7.55 (m, 2H), 7.54 - 7.45 (m, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.89 (d, J = 17.3 Hz, 1H), 5.54 (d, J = 17.3 Hz, 1H), 4.45 (dd, J = 9.1, 5.5 Hz, 1H), 3.98 - 3.87 (m, 1H), 2.42 - 2.28 (m, 1H), 2.28 - 2.12 (m, 1H), 2.00 1.86 (m, 1H), 1.19 - 0.99 (m, 1H), 0.87 - 0.72 (m, 1H); MS (ES+): 583/585 (M+l), (ES-): 581/583 (M-l).

Scheme 299

298b

299a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-phenyl-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (299a)

Compound 299a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-phenyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (298b) (75 mg, 0.173 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (57.6 mg, 0.173 mmol), HATU (79 mg, 0.208 mmol) DIPEA (112 mg, 0.865 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0 - 5%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-6-phenyl-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide

529

(299a) (67 mg, 65% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.79 (s, 1H, D₂O exchangeable), 9.19 - 8.84 (bs, 1H, D₂O exchangeable), 8.98 (d, J = 1.8 Hz, 1H), 8.84 - 8.58 (bs, 1H, D₂O exchangeable), 8.70 (s, 1H), 8.53 (d, J = 1.9 Hz, 1H), 7.99 (d, J = 8.2 Hz, 1H), 7.90 - 7.81 (m, 2H), 7.67 (t, J = 8.0 Hz, 1H), 7.63 - 7.54 (m, 2H), 7.54 - 7.46 (m, 1H), 5 7.30 (d, J = 7.7 Hz, 1H), 5.85 (d, J = 17.3 Hz, 1H), 5.48 (d, J = 17.3 Hz, 1H), 4.40 (dd, J = 9.1,

5.9 Hz, 1H), 3.76 - 3.61 (m, 1H), 2.48 - 2.38 (m, 1H), 1.99 (dd, J = 13.3, 5.8 Hz, 1H), 1.32 (s, 3H), 1.08 - 1.00 (m, 1H), 1.00 - 0.88 (m, 1H); MS (ES+): 597/599 (M+l); (ES-): 595/597 (ΜΙ)·

Scheme 300

F

300a

Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (300a)

Compound 300a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-915 yl)acetic acid (184f) (61.3 mg, 0.140 mmol) in DMF (1.5 mL) using TFA sait of (2S,4R)-N(6-chloropyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (246a) (50 mg, 0.140 mmol), HATU (80 mg, 0.210 mmol), DIPEA (0.122 mL, 0.699 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido [4,5-b] indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-4-fluoropyrrolidine-2carboxamide (300a) (48 mg, 62% yield) HCI sait as a white solid; *H NMR (300 MHz, DMSO-cfe) (a mixture of two rotamers) δ 11.37 and 11.03 (2s, 1H), 8.78 (s, 1H), 8.59 (s, 1H),

530

8.57 - 8.46 (m, 2H), 8.15 and 7.97 (2d, J= 8.2 Hz, 1H), 7.82 (t, J= 8.0 Hz, 1H), 7.59 (s, 1H), 7.31 and 7.19 (2d, J= 7.7 Hz, 1H), 5.89 - 5.42 (m, 3H), 4.68 (t, J= 8.5 Hz, 1H), 4.32 (dd, J= 21.3, 12.6 Hz, 1H), 4.10-3.84 (m, 1H), 2.76 and 2.66 (2s, 3H), 2.61 - 2.53 (m, 1H),2.282.02 (m, 1H); ¹⁹FNMR (282 MHz, DMSO-î/₆) δ -58.73, -176.50; MS (ES+): 550.1 (M+l);

(ES-): 548.1 (M-l).

Scheme 301

184f

Préparation of (2S,4R)-l-(2-(4-amino-8-methyI-6-(trifluoromethyl)-9H-pyrimido[4,510 b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (301a)

Compound 301a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (54.5 mg, 0.124 mmol) in DMF (1.5 mL) using TFA sait of (2S,4R)-N(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (5d) (50 mg, 0.124 mmol), HATU (70.9 mg, 0.187 mmol), DIPEA (0.108 mL, 0.622 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [C18 column (50 g), eluting with ACN in water (containing 0.1% HCI) from 0-100%] (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyI)-9H-pyrimido[4,520 b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yI)-4-fluoropyrrolidine-2-carboxamide (301a) (53 mg, 72% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-îZs) (a mixture of two rotamers) δ 11.38 and 11.04 (2s, 1H, D₂O exchangeable), 8.76 (s, 1H), 8.57 (s, 1H), 8.45 (s, 2H, D₂O exchangeable), 8.18 and 7.99 (2d, J= 8.2 Hz, 1H), 7.84and 7.71 (2t, J= 8.0 Hz, 1H), 7.58 (s, 1H), 7.44 and 7.32 (2d, J= 7.7 Hz, 1H), 5.86 - 5.43 (m, 3H), 4.68 (t, J= 8.5 Hz,

531

1H), 4.31 (dd, 7=21.4, 12.6 Hz, 1 H), 3.98 (dd, 7=38.1, 12.5 Hz, 1H), 2.75 and 2.65 (2s, 3H), 2.61 - 2.54 (m, 1H), 2.30 - 2.02 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-7₆) δ -58.70, 176.49; MS (ES+): 594.1/596.1 (M+l); (ES-): 592.1/594.1 (M-l).

Scheme 302

CF₃

302a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane3-carboxamide (302a)

Compound 302a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (63.2 mg, 0.144 mmol) in DMF (1.5 mL) using HCl sait of (1R,3S,5R)N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (10a) (50 mg, 0.144 mmol), HATU (82 mg, 0.216 mmol), DIPEA (0.126 mL, 0.721 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 -100%] followed by purification using reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-520 methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (302a) (64 mg, 72% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-7₆) δ 10.33 (s, 1H, D₂O exchangeable), 8.73 (s, 1H), 8.49 (s, 1H), 8.13 (s, 2H, D₂O exchangeable), 7.61 (d, 7= 8.0 Hz, 1H), 7.53 (s, 1H), 7.43 (d, 7= 7.9 Hz, 1H), 5.86 (d, 7= 18.0 Hz, 1H), 5.65 (d, J= 17.9 Hz, 1H), 4.37 (dd, 7= 9.1, 5.6 Hz, 1H), 3.70 - 3.65 (m, 1H), 2.73 (s, 3H), 2.60 - 2.57 (m, 1H), 2.08 - 2.02 (m, 1H), 2.01 (s,

532

3H), 1.33 (s, 3H), 1.07 - 1.00 (m, 1H), 0.94 - 0.85 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-J₆) δ -58.58; MS (ES+): 616.2/618.1 (M+l); (ES-): 614.1/616.1 (M-l).

Scheme 303

303a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyI)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3carboxamide (303a)

Compound 303a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-910 yl)acetic acid (184f) (65.9 mg, 0.150 mmol) in DMF (1.5 mL) using HCl sait of (1R,3S,5R)N-(6-bromo-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (Id) (50 mg, 0.150 mmol), HATU (86 mg, 0.225 mmol), DIPEA (0.131 mL, 0.752 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with DMA-80 in DCM from 0 - 100%] followed by purification using reverse phase column chromatography [Cl 8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-2-(2-(4-amino-8-methyl-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (303a) (45 mg, 50% yield) HCl sait as a white solid; ‘HNMR (300 MHz, DMSO-cfc) δ 10.32 (s, 1H, D₂O exchangeable), 8.78 (s, 1H), 8.63 - 8.41 (m, 3H, 2H D₂O exchangeable), 7.63 - 7.50 (m, 2H), 7.42 (d, J= 7.9 Hz, 1H), 5.93 (d, J =

18.3 Hz, 1H), 5.70 (d, J= 17.9 Hz, 1H), 4.38 (dd, J= 9.2, 5.5 Hz, 1H), 3.93 - 3.84 (m, 1H), 2.74 (s, 3H), 2.47-2.34 (m, 1H), 2.30-2.16 (m, 1H), 1.99 (s, 3H), 1.98- 1.88 (m, 1H), 1.16 - 0.99 (m, 1H), 0.82 - 0.64 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -58.69; MS (ES+): 602.1/604.1 (M+l); (ES-): 600.1/602.1 (M-l).

533

Scheme 304

CF₃

184f ^304a

Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoro-4-methyIpyrrolidine-2-carboxamide (304a)

Compound 304a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifhiofomethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (60mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of (2S,4R)-N-(6bromopyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (207a) (57.0 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (2S,4R)-1-(2-(4amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (304a) (41 mg, 49% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 11.36 and 11.02 (2s, 1H, D₂O exchangeable), 8.76 (s, 1H), 8.56 (s, 1H), 8.42 (s, 2H, D₂O exchangeable), 8.18 and 7.99 (2d, J= 8.2 Hz, 1H), 7.84 and 7.71 (2t, J= 8.0 Hz, 1H), 7.58 (s, 1H), 7.43 and 7.32 (2d, J= 7.7 Hz, 1H), 5.76 (d, J= 18.0 Hz, 1H), 5.54 (d, J= 17.9 Hz, 1H), 4.68 (t, J= 8.7 Hz, 1H), 4.41 - 4.20 (m, 1H), 3.96 - 3.81 (m, 1H), 2.75 (s, 3H), 2.62 - 2.56 (m, 1H), 2.23 - 1.95 (m, 1H), 1.62 (d, J= 21.1 Hz, 3H); ¹⁹F NMR (282 MHz, DMSO) δ 58.70, -139.66; MS (ES+): 608.1/610.1 (M+l); (ES-): 606.1/608.1 (M-l); Analysis calculated for C₂₅H₂₂BrF₄N₇O₂ 2H₂O. HCl : C, 44.10; H, 4.00; Cl, 5.21; N, 14.40; Found: C, 43.75; H, 3.89; Cl,5.56; N, 14.11.

Scheme 305

534

Ί 84f 305a

Préparation of (2S,4R)-1 -(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (305a)

Compound 305a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyI)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (60mg, 0.137 mmol) in DMF (1.5 mL) using HCl sait of (2S,4R)-N-(6bromopyrazin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (213c) (46.5 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 10 16 h. This gave after workup and purification as described in scheme-303, (2S,4R)-1-(2-(4amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyrazin-2-yl)-4-fluoro-4-methyIpyrrolidine-2-carboxamide (305a) (64 mg, 77 % yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-ôfc) (a mixture of two rotamers) δ 11.68 and 11.38 (2s, 1H, D₂O exchangeable), 9.40 and 9.22 (2s, 1H), 8.78 (s, 1H), 8.70 - 8.57 (m, 15 3H, 2H D₂O exchangeable), 8.54 (s, 1H), 7.60 (d, J= 1.7 Hz, 1H), 5.79 (d, J= 18.0 Hz, 1H),

5.57 (d, J= 18.0 Hz, 1H), 4.71 (dd, J= 9.8, 7.5 Hz, 1H), 4.30 (dd, J= 18.1, 12.1 Hz, 1H), 3.91 (dd, J= 35.3, 12.1 Hz, 1H), 2.75 (s, 3H), 2.65-2.55 (m, 1H), 2.25 - 1.99 (m, 1H), 1.63 (d, J=21.0 Hz, 3H); ’⁹FNMR(282 MHz, DMSO) δ -58.76, -139.68; MS (ES+): 609.1/611.1 (M+l); (ES-): 607.1/609.1 (M-l); Analysis calculated for C₂₄H₂iBrF₄N₈O₂ 2H₂O. HCl : C,

42.27; H, 3.84; Cl, 5.20; N, 16.43; Found: C, 42.34; H, 3.78; Cl, 4.91; N, 16.16.

Scheme 306

535

184f 306a

Préparation of (S)-l -(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yI)acetyl)-N-(6-bromopyridin-2-yl)pyrrolidine-2-carboxamide (306a)

Compound 306a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (60mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of (S)-N-(6bromopyridin-2-yl)pyrrolidine-2-carboxamide (13a) (52.6 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (S)-l-(2-(4-amino-8-methyl-610 (trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)pyrrolidine2-carboxamide (306a) (76 mg, 96% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-cfc) (a mixture oftwo rotamers) δ 11.28 and 10.94 (2s, 1H, D₂O exchangeable), 8.78 (s, 1H), 8.71 - 8.43 (m, 3H, 2H D₂O exchangeable), 8.15 and 8.00 (2d, J= 8.1 Hz, 1H), 7.82 and 7.71 (2t, J= 8.0 Hz, 1H), 7.60 (d, J= 1.7 Hz, 1H), 7.41 and 7.31 (2d, J= 7.7 Hz, 1H),

5.66 (s, 2H), 4.56 (dd, J= 8.4, 4.5 Hz, 1H), 3.84 (t, J= 6.6 Hz, 2H), 2.77 (s, 3H), 2.33 - 2.16 (m, 1H), 2.13 - 2.00 (m, 2H), 2.00 - 1.85 (m, 1H); ¹⁹F NMR (282 MHz, DMSO) δ -58.74; MS (ES+): 576.1/578.1 (M+l); (ES-): 574.1/576.0 (M-l); Analysis calculated for C₂₄H₂iBrF₃N₇O₂ 1.5 H₂O. HCl: C, 45.05; H, 3.94; Cl, 5.54; N, 15.32; Found: C, 45.19; H, 3.98; Cl, 5.28; N, 15.22.

Scheme 307

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(pyridin-4-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3 -carboxamide (307c)

Step-1 : Préparation of ZerZ-butyl 2-(4-amino-6-(pyridin-4-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yI)acetate (307a)

Compound 307a was prepared according to the procedure reported in step-1 of scheme-263, from ZerZ-butyl 2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (290e) (300 mg, 0.793 mmol) in rc-BuOH (4 mL) using pyridin-4-ylboronic acid (146 mg,

1.190 mmol), Pd₂(dba)₃ (72.6 mg, 0.079 mmol), XPhos (76 mg, 0.159 mmol), a solution of

1.27 M potassium phosphate (1.249 mL, 1.586 mmol) and heating at 100 °C for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0-7%] ZerZ-butyl 2-(4-amino-6-(pyridin-4-yl)-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidm-9-yl)acetate (307a) (275 mg, 92% yield) as a gray solid; ‘HNMR (300 MHz, DMSO-J₆) δ 8.99 (d, J= 1.8 Hz, 1H), 8.73 (d, 2H), 8.61 (d, J = 1.9 Hz, 1H), 8.41 (s, 1H), 8.01 (s, 1H), 7.91 (d, 2H), 6.89 (bs, 1H), 5.25 (s, 2H), 1.41 (s, 9H); MS (ES+): 377 (M+l).

Step-2: Préparation of 2-(4-amino-6-(pyridin-4-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetic acid (307b)

Compound 307b was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6-(pyridin-4-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-921436

537 yl)acetate (307a) (272 mg, 0.723 mmol) in DCM (5 mL) using TFA (824 mg, 7.23 mmol) and stirring at RT for 16 h. This gave after work up 2-(4-amino-6-(pyridin-4-yl)-9Hpyrido[2^,,3':4,5]pyrrolo[2,3-d]pyrimidin-9-yI)acetic acid (307b) (358 mg) TFA sait as a beige solid; Ή NMR (300 MHz, DMSO-riô) δ 9.28 (d, J= 1.9 Hz, 1H), 9.13 - 9.03 (m, 3H), 5 8.97 (bs, 1H), 8.68 (s, 1H), 8.60 (d, 2H), 8.30 (bs, 1H), 5.39 (s, 2H); MS (ES+) 321 (M+l);

(ES-): 319.10 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(pyridin-4-yl)-9Hpyrido[2',3':4,5]pynOlo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (307c)

Compound 307c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(pyridin-4-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (307b) (75 mg, 0.173 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (55.0 mg, 0.173 mmol), HATU (79 mg, 0.207 mmol), DIPEA (112 mg, 0.863 mmol) and stirring at RT for 16

h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4amino-6-(pyridin-4-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (307c) (52 mg, 52% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ 10.78 (s, 1H, D₂O exchangeable), 9.20 (d, J = 1.9 Hz, 1H), 9.02 (d, J = 5.9 Hz, 2H), 8.86 - 8.68 (bs, 1H, D₂O exchangeable),

8.77 (s, 1H), 8.63 (s, 1H), 8.46 (d, J = 6.0 Hz, 2H), 8.13 - 7.78 (bs, 1H, D₂O exchangeable),

7.97 (d, J = 8.2 Hz, 1H), 7.68 (t, J = 8.0 Hz, 1H), 7.30 (d, J = 7.7 Hz, 1H), 5.85 (d, J = 17.2 Hz, 1H), 5.53 (d, J = 17.2 Hz, 1H), 4.49 - 4.41 (m, 1H), 3.95 - 3.87 (m, 1H), 2.39 - 2.29 (m, 1H), 2.29 - 2.17 (m, 1H), 2.01 - 1.81 (m, 1H), 1.20 - 1.04 (m, 1H), 0.90 - 0.74 (m, 1H); MS (ES+): 584/586 (M+l); (ES-): 582/584 (M-l); Analysis calculated for

C₂₇H₂₂BrN₉O₂.2.2HC1.4.5H₂O: C, 43.49; H, 4.49; Cl, 10.46; N, 16.90; Found: C, 43.57; H,

4.29; Cl, 10.41; N, 16.69.

Scheme 308

I

538

I !

ί ι

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (308c)

Step-1 : Préparation of /eri-butyl 2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetate (308a)

Compound 308a was prepared according to the procedure reported in step-1 of scheme-263, from tert-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (192e) (500 mg, 1.278 mmol) in n-BuOH (20 mL) using (2-methylpyrimidin-5-yl)boronic acid (264 mg, 1.917 mmol), Pd₃(dba)₃ (117 mg, 0.128 mmol), XPhos (122 mg, 0.256 mmol), a solution of 1.27 M potassium phosphate (2.012 mL, 2.56 mmol) and heating at 100 °C for 16 h. This gave after workup and purification by flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0-7%] Zert-butyl 2-(4-amino-8-methyl-6-(2-methylpyrimidin-5yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (308a) (475 mg, 92% yield) as a white solid; Ή

NMR (300 MHz, DMSO-îZ₆) δ 9.21 (s, 2H), 8.59 (d, J= 1.8 Hz, 1H), 8.33 (s, 1H), 7.64 (s, 1H), 7.49 (s, 2H), 5.37 (s, 2H), 2.71 (s, 3H), 2.68 (s, 3H), 1.44 (s, 9H); MS (ES+): 405 (M+l); (ES-): 403 (M-l).

Step-2: Préparation of 2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (308b)

539

Compound 308b was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5-b]indol-9yl)acetate (308a) (475 mg, 1.174 mmol) in DCM (20 mL) using TFA (1339 mg, 11.74 mmol) and stirring at RT for 16 h. This gave after work up TFA sait 2-(4-amino-8-methyl-6-(25 methylpyrimidin-5-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (308b) (715 mg) as a pale yellow solid; Ή NMR (300 MHz, DMSO-ri₆) δ 9.21 (s, 2H), 8.82 (bs, 2H), 8.73 (d, J= 1.8 Hz, 1H), 8.65 (s, 1H), 7.80 (s, 1H), 5.48 (s, 2H), 2.77 (s, 3H), 2.70 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-de) δ -74.78; MS (ES+): 349 (M+l); (ES-): 347 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H10 pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (308c)

Compound 308c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yI)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (308b) (75 mg, 0.162 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (51.7 mg, 0.162 mmol), HATU (74.0 mg, 0.195 mmol), DIPEA (105 mg, 0.81 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (308c) (70 mg, 71% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.84 (s, 1H, D₂O exchangeable), 9.23 (s, 2H), 9.06 (bs, 2H, D₂O exchangeable), 8.77 (d, J = 1.7 Hz, 1H), 8.69 (s, 1H), 8.00 (d, J = 8.2 Hz, 1H), 7.79 (s, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.97 (d, J = 18.0 Hz, 1H), 5.67 (d, J = 17.9 Hz, 1H), 4.43 (dd, J = 9.1, 5.7 Hz, 1H), 3.98 - 3.91 (m, 1H), 2.78 (s, 3H), 2.69 (s, 3H), 2.42 - 2.29 (m, 1H), 2.29 - 2.10 (m, 1H), 2.02

- 1.87 (m, 1H), 1.17 - 1.02 (m, 1H), 0.79 - 0.62 (m, 1H); MS (ES+): 612/614 (M+l); (ES-):

610/612 (M-l); Analysis calculated for C₂9H₂₆BrN₉O₂.1.5HC1.4H₂O: C, 47.12; H, 4.84; Cl, 7.19; N, 17.05; Found: C, 47.26; H, 4.67; Cl, 7.17; N, 17.07.

Scheme 309

540

308b 309a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9Hpyrimido [4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (309a)

Compound 309a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (308b) (75 mg, 0.162 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (54 mg, 0.162 mmol), HATU (74.0 mg, 0.195 mmol), DIPEA (105 mg, 0.811 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yI)-5-methyI-2-azabicyclo[3.1.0]hexane-3carboxamide (309a) (64 mg, 63% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-dg) δ 10.84 (s, 1H, D₂O exchangeable), 9.23 (s, 1H), 9.21 (s, 1H), 9.01 (bs, 1H, D₂O exchangeable), 8.89 (bs, 1H, D₂O exchangeable), 8.76 (s, 1H), 8.71 - 8.64 (m, 1H), 8.02 (d, J = 8.2 Hz, 1H), 7.79 (s, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.92 (d, J = 18.0 Hz, 1H), 5.63 (d, J = 17.8 Hz, 1H), 4.39 (dd, J = 9.0, 6.2 Hz, 1H), 3.75 - 3.70 (m, 1H), 2.77 (s, 3H), 2.69 (s, 3H), 2.49 - 2.44 (m, 1H), 1.99 (dd, J= 13.3, 6.1 Hz, 1H), 1.32 (s, 3H), 1.11 - 0.99 (m, 1H), 0.92 - 0.81 (m, 1H); MS (ES+) 626/628 (M+l); (ES-) 624/626 (M-l);

Analysis calculated for C₃oH₂8BrN₉0₂.1.6HC1.4H₂0: C, 47.60; H, 5.01; Cl, 7.49; N, 16.65; Found: C, 47.59; H, 4.89; Cl, 7.36; N, 16.44.

Scheme 310

541

308b 310a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9Hpyrimido [4,5-b] indol-9-yl)acetyl)-N-(6-bromo-3 -methylpyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (310a)

Compound 310a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (308b) (75 mg, 0.162 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromo-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (Id) (54 mg, 0.162 mmol), HATU (74 mg, 0.195 mmol), DIPEA (105 mg, 0.811 mmol) and 10 stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, ( 1 R,3 S,5R)-2-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5 -yl)-9H-pyrimido[4,5-b] indol-9yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (310a) (41 mg, 40% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-cfe) δ 10.35 (s, 1H, D2O exchangeable), 9.26 (s, 2H), 9.11 (bs, 2H, D2O exchangeable), 8.80 (s, 1H), 8.70 15 (s, 1H), 7.79 (s, 1H), 7.60 (d, J = 8.0 Hz, 1H), 7.42 (d, J = 7.9 Hz, 1H), 5.96 (d, J = 18.0 Hz,

1H), 5.68 (d, J = 17.8 Hz, 1H), 4.41 (dd, J = 9.2, 5.6 Hz, 1H), 3.95 - 3.93 (m, 1H), 2.76 (s, 3H), 2.71 (s, 3H), 2.48 - 2.36 (m, 1H), 2.32 - 2.19 (m, 1H), 2.00 (s, 3H), 1.98 - 1.91 (m, 1H), 1.19 - 0.98 (m, 1H), 0.89 - 0.65 (m, 1H); MS (ES+) 626/628 (M+l); (ES-) 624/626 (M-l); Analysis calculated for C3oH₂8BrN₉02.1.5HC1.5.25H20: C, 46.45; H, 5.20; N, 16.25; Found:

C, 46.48; H, 4.98; N, 16.02.

Scheme 311

542

308b 311a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (311a)

Compound 311a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (308b) (75 mg, 0.162 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (10a) (56.2 mg, 0.162 mmol), HATU (74.0 mg, 0.195 mmol), DIPEA (105 mg,

0.811 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (311a) (61 mg, 59% yield) HCl sait as a white solid; ‘H NMR (300 MHz, DMSO-J₆) δ 10.37 (s, 1H, D₂O exchangeable), 9.24 (s, 2H), 9.04 (bs,

2H, D₂O exchangeable), 8.78 (d, J = 1.7 Hz, 1H), 8.68 (s, 1H), 7.78 (s, 1H), 7.61 (d, J = 8.0

Hz, 1H), 7.42 (d, J = 7.9 Hz, 1H), 5.90 (d, J = 18.0 Hz, 1H), 5.63 (d, J = 17.9 Hz, 1H), 4.39 (dd, J = 9.1, 5.7 Hz, 1H), 3.71 - 3.68 (m, 1H), 2.74 (s, 3H), 2.70 (s, 3H), 2.63 - 2.53 (m, 1H), 2.13 - 2.04 (m, 1H), 2.02 (s, 3H), 1.34 (s, 3H), 1.10 - 0.97 (m, 1H), 0.98 - 0.85 (m, 1H); MS (ES+): 640/642 (M+l), (ES-): 638/640 (M-l); Analysis calculated for

C₃iH3oBrN₉0₂.1.5HC1.3.5H₂0: C, 49.10; H, 5.12; Cl, 7.01; N, 16.62; Found: C, 48.93; H,

4.95; Cl, 7.05; N, 16.39.

Scheme 312

543

Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2-. carboxamide (312a)

Compound 312a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(2-methyIpyrimidin-5-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (308b) (75 mg, 0.162 mmol) in DMF (5 mL) using HCl sait of (2S,4R)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (5d) (52.6 mg, 0.162 mmol), HATU (74.0 mg, 0.195 mmol), DIPEA (105 mg, 0.811 mmol) and stirring at RT for 10 16 h. This gave after workup and purification as described in scheme-299, (2S,4R)-1-(2-(4amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (312a) (26 mg, 26% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-dô) δ (a mixture of two rotamers) 11.39 and 11.05 (2s, 1H, D2O exchangeable), 9.23 and 9.21 (2s, 2H), 8.94 (s, 2H, D2O exchangeable), 15 8.76 (d, J= 1.7 Hz, 1H), 8.67 and 8.65 (2s, 1H), 8.18 and 7.99 (2d, J= 8.2 Hz, 1H), 7.82 and

7.70 (2t, J= 8.0 Hz, 1H), 7.79 (s, 1H), 7.44 and 7.31 (d, J= 7.7 Hz, 1H), 5.91 - 5.09 (m, 3H), 4.69 (t, J= 8.5 Hz, 1H), 4.42 - 4.22 (m, 1H), 4.10 - 3.93 (m, 1H), 2.77 (s, 3H), 2.69 (s, 3H), 2.32-2.16 (m, 1H), 2.16-2.01 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-Jô) δ-176.18; MS (ES+): 618/620 (M+l); (ES-): 616/618 (M-l).

Scheme 313

544

CF₃ 184f 313b

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yI)acetyl)-N-(6-bromo-5-methylpyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (313b)

Compound 313b was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (75 mg, 0.171 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)-N(6-bromo-5-methylpyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (313a) (57.1 mg, 0.171 mmol; CAS # 2086190-99-8), HATU (98 mg, 0.257 mmol), DIPEA (111 mg, 0.856 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromo-5-methylpyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (313b) (52 mg, 50% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-îZô) δ 11.08 (s, 1H, D₂O exchangeable), 9.10 (s, 1H), 8.80 (s, 1H), 8.71 (bs, 2H, D₂O exchangeable), 8.65 (s, 1H), 7.61 (s, 1H), 5.98 (d, J = 18.1 Hz, 1H), 5.70 (d, J = 17.9 Hz, 1H), 4.44 (dd, J = 9.1, 5.6 Hz, 1H), 3.98 - 3.90 (m, 1H), 2.77 (s, 3H), 2.52 (s, 3H), 2.43 - 2.29 (m, 1H), 2.29 - 2.14 (m, 1H), 2.01 - 1.87 (m, 1H), 1.17 - 0.99 (m, 1H), 0.82 - 0.62 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-^) δ -58.78; MS (ES+): 603/605 (M+l), (ES-): 601/603 (M-l);

Analysis calculated for C₂5H₂₂BrF₃N8O₂.1.4HC1.1.5H₂O: C, 44.06; H, 3.90; N, 16.44; Found: 20 C, 44.02; H, 3.55; N, 16.44.

Scheme 314

545

189f 314a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-9yI)acetyl)-N-(6-bromopyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (314a)

Compound 314a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (189f) (75 mg, 0.195 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyrazin-2yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (261c) (65.1 mg, 0.195 mmol), HATU (111 mg, 0.293 mmol), DIPEA (126 mg, 0.976 mmol) and stirring at RT for 16 h.

This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4amino-6,8-dimethyl-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-bromopyrazin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (314a) (66 mg, 62% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-cfo) δ 11.20 (s, 1H, D₂O exchangeable), 9.24 (s, 1H), 8.64 (bs, 2H, D₂O exchangeable), 8.61 (s, 1H), 8.54 (s, 1H), 8.17 (s, 1H), 7.12 (s, 1H), 5.86 (d, J = 18.0 Hz, 1H), 5.58 (d, J = 17.9 Hz, 1H), 4.41 (dd, J = 9.1, 6.1 Hz, 1H), 3.74 - 3.69 (m,

1H), 2.65 (s, 3H), 2.50 - 2.45 (m, 1H), 2.43 (s, 3H), 2.12 - 1.98 (m, 1H), 1.31 (s, 3H), 1.09 0.95 (m, 1H), 0.92 - 0.81 (m, 1H); MS (ES+): 549/551 (M+l); (ES-): 547/549 (M-l);

Analysis calculated for C₂₅H₂₅BrN₈O₂.l.lHCl. 2H₂O: C, 48.00; H, 4.85; Cl, 6.23; N, 17.91; Found: C, 47.84; H, 4.67; Cl, 6.10; N, 17.90.

Scheme315

546

Me

308b 315a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (315a)

Compound 315a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5-b]indol9-yl)acetic acid (308b) (75 mg, 0.162 mmol) in DMF (5 mL) using HCl sait of (1R,3S,5R)N-(6-chloropyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (269b) (46.7 mg, 0.162 mmol), HATU (74.0 mg, 0.195 mmol), DIPEA (105 mg, 0.811 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-chloropyridin-2-yl)-5-methyl-2-azabicyclo[3.1,0]hexane-3-carboxamide (315a) (47 mg, 50% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.83 (s, 1H, D₂O exchangeable), 9.24 (s, 2H), 9.06 (bs, 2H, D₂O exchangeable), 8.77 (d, J = 1.7

Hz, 1H), 8.69 (s, 1H), 7.99 (d, J = 8.2 Hz, 1H), 7.87 - 7.74 (m, 2H), 7.18 (d, J = 7.7 Hz, 1H), 5.92 (d, J = 18.0 Hz, 1H), 5.62 (d, J = 17.9 Hz, 1H), 4.40 (dd, J = 9.0, 6.2 Hz, 1H), 3.76 - 3.70 (m, 1H), 2.76 (s, 3H), 2.70 (s, 3H), 2.49 - 2.43 (m, 1H), 1.99 (dd, J = 13.3, 6.1 Hz, 1H), 1.32 (s, 3H), 1.08 - 0.95 (m, 1H), 0.94 - 0.76 (m, 1H); MS (ES+): 582/584 (M+l); (ES-): 580/582 (M-l); Analysis calculated for C₃₀H₂₈ClN₉O₂.1.3HCL3H₂O: C, 52.72; H, 5.21; Cl, 11.93; N,

18.44; Found: C, 52.74; H, 5.19; Cl, 11.84; N, 18.36.

Scheme 316

547

Me 308b 316a

Préparation of (2S,4R)-1 -(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9Hpyrimido[4,5-b]indol-9-yI)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoro-4-niethylpyrroIidine-2carboxamide (316a)

Compound 316a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5-b]indol9-yl)acetic acid (308b) (75 mg, 0.162 mmol) in DMF (5 mL) using TFA sait of (2S,4R)-N(6-bromopyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (207a) (67.5 mg, 0.162 mmol), HATU (74.0 mg, 0.195 mmol), DIPEA (105 mg, 0.811 mmol) and stirring at RT for

16 h. This gave after workup and purification as described in scheme-299, (2S,4R)-1-(2-(4amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (316a) (60 mg, 59% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-îZô) δ (a mixture of two rotamers) 11.37 and 11.03 (2s, 1H, D₂O exchangeable), 9.22 (s, 2H), 8.93 (bs, 2H, D2O exchangeable), 8.77 (s, 1H), 8.67 and 8.65 (2s, 1H), 8.19 and 7.99 (2d, J = 8.2 Hz, 1H), 7.79 (d, J = 1.6 Hz, 1H),

7.85 and 7.70 (2t, J = 8.0 Hz, 1H), 7.44 and 7.32 (2d, J = 7.7 Hz, 1H), 5.77 (d, J = 18.0 Hz, 1H), 5.56 (d, J = 17.9 Hz, 1H), 4.70 (dd, J = 9.7, 7.5 Hz, 1H), 4.29 (dd, J = 18.2, 12.3 Hz, 1H), 3.99 - 3.81 (m, 1H), 2.77 (s, 3H), 2.69 (s, 3H), 2.63 - 2.55 (m, 1H), 2.24 - 1.96 (m, 1H), 1.63 (d, J = 21.1 Hz, 3H); ¹⁹F NMR (282 MHz, DMSO-i/g) δ -139.89; MS (ES+): 632/634 (M+l); (ES-): 630/632 (M-l); Analysis calculated for C29H27BrFN₉O2.1.55HC1.2.5H2O: C,

47.45; H, 4.61; Cl, 7.49; N, 17.17; Found: C, 47.48; H, 4.71; Cl, 7.45; N, 17.11.

Scheme 317

548

cf₃ 317c

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyI)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromo-3-fluoropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (317c)

Step-1 : Préparation of (lR,3S,5R)-ZerZ-butyl 3-((6-bromo-3-fluoropyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (317a)

Compound 317a was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(ZerZ-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (259a) (0.5 g, 2.2mmol) and 6-bromo-3-fluoropyridin-2-amine (0.42 g, 2.2 mmol; CAS # 10 1379457-78-9) to afford after purification using flash column chromatography [silica gel (24

g), EtOAc in hexane from 0 - 50%] (1 R,3S,5R)-ZerZ-butyl 3-((6-bromo-3-fluoropyridin-2' yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (317a) (180 mg, 20% yield) as a white solid; MS (ES+): 400.1/402.1(M+l).

Step-2: Préparation of (lR,3S,5R)-N-(6-bromo-3-fluoropyridin-2-yl)-215 azabicyclo[3.1.0]hexane-3-carboxamide (317b)

Compound 317b was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-ZerZ-butyl 3-((6-bromo-3-fluoropyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (317a) (0.18 g, 0.450 mmol) in DCM (2.5 mL) using

TFA (0.243 mL, 3.15 mmol). This gave after work up TFA sait of (lR,3S,5R)-N-(6-bromo20 3-fluoropyridin-2-yl)-2-azabicycIo[3.1.0]hexane-3-carboxamide (317b) (180 mg, 97% yield);

’H NMR (300 MHz, DMSO-cfc) δ 10.95 (d, J= 13.0 Hz, 1H), 8.50 (dd, J= 4.5, 2.5 Hz, 1H),

549

8.32 (d, J= 5.7 Hz, 1H), 4.30 - 4.18 (m, 1H), 3.41 - 3.31 (m, 1H), 2.66 - 2.57 (m, 1H), 2.34 - 2.09 (m, 1H), 1.92 - 1.70 (m, 1H), 0.92 - 0.73 (m, 2H); ¹⁹F NMR (282 MHz, DMSO-î/ô) δ -73.77, -142.14; MS (ES+): 300.0/302.0 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H5 pyrimido [4,5-b] indol-9-yl)acetyI)-N-(6-bromo-3-fluoropyridin-2-yl)-2azabicycio [3.1.0]hexane-3 -carboxamide (317c)

Compound 317c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (lR,3S,5R)-N-(6-bromo-3-fluoropyridin-2-yl)-2-azabicyclo[3.1.0]hexane3-carboxamide (317b) (56.7 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(410 amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (60mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromo-3-fluoropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (317c) (47mg, 57% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ 10.49 (s, 1H, D₂O exchangeable), 8.78 (s, 1H), 8.59 (s, 1H), 8.50 (s, 2H, D2O exchangeable), 8.37 (d, J= 2.5 Hz, 1H), 8.34 (d, J= 5.7 Hz, 1H), 7.61 (s, 1H), 5.97 (d, J= 18.0 Hz, 1H), 5.70 (d, J= 18.0 Hz, 1H), 4.59 (dd, J = 8.9, 5.8 Hz, 1H), 4.01-3.89 (m, 1H), 2.78 (s, 3H), 2.422.20 (m, 2H), 2.02 - 1.86 (m, 1H), 1.15 - 1.05 (m, 1H), 0.77 - 0.68 (m, 1H); ¹⁹F NMR (282

MHz, DMSO-î/ô) δ -58.71, -142.98; MS (ES+): 606.1/608.1 (M+l); (ES-): 604.1/606.1 (ΜΙ); Analysis calculated for C25H2oBrF4N₇0₂ 2H₂O. HCl.: C, 44.23; H, 3.71; Cl, 5.22; N, 14.44; Found: C, 44.06; H, 3.73; Cl,4.96; N, 14.31.

Scheme 318

550

HATU, DIPEA

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b] indol-9-y l)acetyl)-N-(6-chloro-3 -methy lpyridin-2-y l)-2-azabicy cio [3.1.0] hexane-3 carboxamide (318c)

Step-1 : Préparation of (lR,3S,5R)-ierZ-butyl 3-((6-chloro-3-methylpyridin-2-yl)carbamoyl)2-azabicyclo[3.1.0]hexane-2-carboxylate (318a)

Compound 318a was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(terributoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (259a) (0.5 g, 2.2 mmol) in DCM (15 mL) and 6-chloro-3-methylpyridin-2-amine (0.314 g, 10 2.200 mmol; CAS # 442128-86-1) to afford after purification as described in step-1 of scheme-317 (lR,3S,5R)-fôrributyl 3-((6-chIoro-3-methylpyridin-2-yI)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (318a) (0.46 g, 59% yield) as a white solid; MS (ES+): 352.1(M+1).

Step-2: Préparation of (lR,3S,5R)-N-(6-chloro-3-methylpyridin-2-yl)-215 azabicyclo[3.1.0]hexane-3-carboxamide (318b)

Compound 318b was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-teri-butyl 3-((6-chloro-3-methylpyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (318a) (0.45 g, 1.279 mmol) in DCM (7 mL) using TFA (0.690 mL, 8.95 mmol). This gave after work up TFA sait of (lR,3S,5R)-N-(6-chloro-320 methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (318b) (0.47 g); 'H NMR (300 MHz, DMSO-î/ô) δ 10.79 (s, 1H), 9.86 (s, 1H), 9.23 (s, 1H), 7.83 (d, J= 8.0 Hz, 1H), 7.42 (d, J= 8.0 Hz, 1H), 4.34 - 4.15 (m, 1H), 3.37 - 3.26 (m, 1H), 2.68 - 2.58 (m, 1H), 2.15

551 (s, 3H), 2.15-2.05 (m, 1H), 1.95- 1.75 (m, 1H), 0.97-0.78 (m, 2H); MS (ES+): 252.0 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido [4,5-b] indol-9-yl)acetyl)-N-(6-chloro-3 -methylpyridin-2-y l)-2azabicyclo[3.1.0]hexane-3-carboxamide (318c)

Compound 318c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (lR,3S,5R)-N-(6-chloro-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane3-carboxamide (318b) (50.1 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(4amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (60mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indoI-9yI)acetyl)-N-(6-chloro-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (318c) (19 mg, 25% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ 10.30 (s, 1H, D₂O exchangeable), 8.82 (s, 1H), 8.73 (s, 2H, D2O exchangeable), 8.65 (d, J= 1.7 Hz, 1H), 7.71 (d, J= 8.0 Hz, 1H), 7.61 (d, J= 1.7 Hz, 1H), 7.30 (d, J= 7.9 Hz, 1H), 5.96 (d, J= 18.0 Hz, 1H), 5.72 (d, J= 17.9 Hz, 1H), 4.40 (dd, J= 9.2, 5.5 Hz, 1H), 3.99 - 3.88 (m, 1H), 2.76 (s, 3H), 2.42 (dd, J= 13.4, 9.3 Hz, 1H), 2.33-2.19 (m, 1H), 2.02 (s, 3H), 2.01 - 1.90 (m, 1H), 1.17 - 1.03 (m, 1H), 0.82 - 0.66 (m, 1H); ¹⁹F NMR (282 MHz, DMSO) δ -58.69; MS (ES+): 558.2 (M+l); (ES-): 556.1 (M-l).

Scheme 319

552

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloro-3-fluoropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (319c)

Step-1: Préparation of (lR,3S,5R)-tert-butyI 3-((6-chloro-3-fluoropyridin-2-yl)carbamoyl)-25 azabicyclo[3.1.0]hexane-2-carboxylate (319a)

Compound 319a was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(Zer/-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (259a) (0.25 g, 1.1 mmol) and 6-chloro-3-fluoropyridin-2-amine (0.161 g, 1.1 mmol; CAS # 1260672-14-7) to afford after purification as described in step-1 of scheme-317 (1R,3S,5R)10 teri-butyl 3-((6-chloro-3-fluoropyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2carboxylate (319a) (0.187 g, 48% yield) as a white solid; MS (ES+): 356.1 (M+l); 378.1 (M+Na).

Step-2: Préparation of (lR,3S,5R)-N-(6-chloro-3-fluoropyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (319b)

Compound 319b was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-Zer/-butyl 3-((6-chloro-3-fluoropyridin-2-yl)càrbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (319a) (0.18 g, 0.506 mmol) in DCM (2.7 mL) using TFA (0.273 mL, 3.54 mmol). This gave after work up and purification using reverse phase column chromatography [Cl 8 column (50 g), eluting with ACN in water (containing 0.1%

HCl) from 0-100%] TFA/HC1 sait of (lR,3S,5R)-N-(6-chloro-3-fluoropyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (319b) (0.19 g); ^JH NMR (300 MHz, DMSO-cfc) δ 11.11 (s, 1H), 9.82 (s, 1 H), 9.29 (s, 1H), 7.99 (t, 7= 8.9 Hz, 1H), 7.56 (dd, 7= 8.6, 3.0 Hz, 1H), 4.30-4.23 (m, 1H), 3.38 - 3.28 (m, 1H), 2.70-2.56 (m, 1 H), 2.15 - 2.02 (m, 1H), 1.94 - 1.71 (m, 1H), 1.04 - 0.70 (m, 2H); MS (ES+): 256.1 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido [4,5-b] indol-9-yl)acetyl)-N-(6-chloro-3 -fluoropyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (319c)

Compound 319c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (lR,3S,5R)-N-(6-chloro-3-fluoropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-330 carboxamide (319b) (50.6 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(4-amino8-methyl-6-(trifhioromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (60mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(2(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloro-3

Ί

553 fluoropyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (319c) (27 mg, 35.1 % yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.56 (s, 1H, D₂O exchangeable), 8.81 (s, 1H), 8.68 (s, 2H, D₂O exchangeable), 8.64 (s, 1H), 7.86 (t, J= 8.9 Hz, 1H), 7.60 (s, 1H), 7.43 (dd, J= 8.5, 2.9 Hz, 1H), 5.96 (d, J= 18.0 Hz, 1H), 5.70 (d, J= 17.9 Hz, 1H), 4.44 (dd, J= 9.3, 5.3 Hz, 1H), 3.95 - 3.90 (m, 1H), 2.76 (s, 3H), 2.42 (dd, J= 13.5, 9.4 Hz, 1H), 2.30-2.16 (m, 1H), 2.03-1.90 (m, 1H), 1.19-1.04 (m, 1H), 0.77-0.69 (m, 1H); ¹⁹FNMR (282 MHz, DMSO) δ -58.75, -125.59; MS (ES+): 562.2 (M+l); (ES-): 560.1 (M-l).

Scheme 320

Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (320c)

Step-1: Préparation of (2S,4R)-ZerZ-butyl 2-((6-chloropyridin-2-yl)carbamoyl)-4-fluoro-4methylpyrrolidine-1 -carboxylate (320a)

Compound 320a was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4R)-l-(ZerZ-butoxycarbonyl)-4-fluoro-4-methylpyrrolidine-2-carboxylic acid (213a) (0.35 g, 1.415 mmol) and 6-chloropyridin-2-amine (0.182 g, 1.415 mmol; CAS # 45644-211) to afford after purification as described in step-1 of scheme-317, (2S,4R)-ZerZ-butyl 2-((6chloropyridin-2-yl)carbamoyl)-4-fluoro-4-methylpyrrolidine-l-carboxylate (320a) (0.362 g, 72% yield) as a white solid; MS (ES+): 358.2 (M+l); 380.1 (M+Na).

Step-2: Préparation of (2S,4R)-N-(6-chloropyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2carboxamide (320b)

554

Compound 320b was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-terZ-butyl 2-((6-chloropyridin-2-yl)carbamoyl)-4-fluoro-4-methylpyrrolidine-1 carboxylate (320a) (0.36 g, 1.006 mmol) in DCM (5.5 mL) using TFA (0.543 mL, 7.04 mmol) to afford after work up TFA sait of (2S,4R)-N-(6-chloropyridin-2-yl)-4-fluoro-45 methylpyrrolidine-2-carboxamide (320b) (0.375 g); ’H NMR (300 MHz, DMSO-àfc) δ 11.45 (s, 1H), 9.99 (s, 1H), 9.12 (s, 1H), 8.05 (d, J= 8.2 Hz, 1H), 7.95 (t, J= 7.9 Hz, 1H), 7.33 (d, J = Ί.Ί Hz, 1H), 4.76-4.51 (m, 1H), 3.68 - 3.29 (m, 2H), 2.88 - 2.67 (m, 1H), 2.35-2.11 (m, 1H), 1.55 (dd, J= 21.3, 5.8 Hz, 3H); MS (ES+): 258.1 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,510 b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (320c)

Compound 320c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (2S,4R)-N-(6-chloropyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2carboxamide (320b) (50.9 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(4-amino15 8-methyl-6-(trifluoromethyI)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (2S,4R)-1-(2-(4amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6chloropyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (320c) (46mg, 60% yield)

HCl sait as a white solid; ’H NMR (300 MHz, DMSO-Jô) (a mixture of two rotamers) δ 11.34 and 11.00 (2s, 1H, D₂O exchangeable), 8.76 (s, 1H), 8.57 (s, 1H), 8.45 (s, 2H, D₂O exchangeable), 8.16 and 7.97 (2d, J= 8.2 Hz, 1H), 7.82 (t, J= 8.0 Hz, 1H), 7.58 (s, 1H), 7.30 and 7.19 (2d, J= 7.7 Hz, 1H), 5.76 (d, J= 18.0 Hz, 1H), 5.54 (d, J= 17.9 Hz, 1H), 4.68 (t, J = 8.6 Hz, 1H), 4.28 (dd, J= 18.0, 12.0 Hz, 1H), 3.88 (dd, J= 35.2, 12.0 Hz, 1H), 2.75 (s,

3H), 2.62-2.54 (m, 1H), 2.23 - 1.94 (m, 1H), 1.63 (d, J=21.1 Hz, 3H); ’⁹F NMR (282 MHz,

DMSO-dô) δ -58.71, -139.94; MS (ES+): 564.2 (M+l); (ES-): 562.1 (M-l); Analysis calculated for C₂₅H₂₂C1F4N₇O₂2H₂O. 0.9HC1.: C, 47.45; H, 4.28; Cl, 10.65; N, 15.49; Found: C, 47.18; H, 3.93; Cl,10.40; N, 15.20.

Scheme 321

Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indoI-9-yl)acetyI)-N-(6-bromopyrazin-2-yl)-4-fluoropyrrolidine-2-carboxamide (321d) Step-1: Préparation of (2S,4R)-/er/-butyl 2-((6-bromopyrazin-2-yl)carbamoyl)-45 fluoropyrrolidine-l-carboxylate (321b)

Compound 321b was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4R)-l-(fôrF-butoxycarbonyl)-4-fluoropyrrolidine-2-carboxylic acid (321a) (0.5 g, 2.144 mmol; CAS # 203866-14-2) and 6-bromopyrazin-2-amine (0.373 g, 2.144 mmol; CAS # 54237-53-5) to afford after purification as described in step-1 of scheme-317, (2S,4R)-fërA butyl 2-((6-bromopyrazin-2-yl)carbamoyI)-4-fluoropyrrolidine-l-carboxylate (321b) (0.452 g, 54% yield) as a white solid; MS (ES+): 389.1/391.1(M+1).

Step-2: Préparation of (2S,4R)-N-(6-bromopyrazin-2-yl)-4-fluoropyrrolidine-2-carboxamide (321c)

Compound 321c was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-terAbutyl 2-((6-bromopyrazin-2-yl)carbamoyl)-4-fluoropyrrolidine-lcarboxylate (321b) (0.45 g, 1.156 mmol) in DCM (6 mL) using TFA (0.624 mL, 8.09 mmol) to afford after work up TFA sait of (2S,4R)-N-(6-bromopyrazin-2-yl)-4-fluoropyrrolidine-2carboxamide (321c) (0.569 g); ’H NMR (300 MHz, DMSO-î/₆) δ 11.80 (s, 1H), 9.30 (s, 1H), 8.67 (s, 1H), 5.65 - 5.37 (m, 1H), 4.66 (dd, J= 10.8, 7.4 Hz, 1H), 3.71 - 3.47 (m, 2H), 2.88 20 2.66 (m, 1H), 2.44 - 2.18 (m, 1H); MS (ES+): 289.0/290.9 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-4-fluoropyrrolidine-2-carboxamide (321d)

556

Compound 321d was prepared according to the procedure reported m step-3 of scheme-1, from TFA sait of (2S,4R)-N-(6-bromopyrazin-2-yl)-4-fluoropyrrolidine-2-carboxamide (321c) (70.8 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(4-amino-8-methyl-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (2S,4R)-l-(2-(4-amino8-methyl-6-(trifluoromethyI)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyrazin-2yl)-4-fluoropyrrolidine-2-carboxamide (321d) (45 mg, 55% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 11.69 and 11.39 (2s, 1H, D2O exchangeable), 9.40 and 9.22 (2s, 1H), 8.77 (s, 1H), 8.65 and 8.58 (2s, 1H), 8.54 (s, 1H), 8.48 (s, 2H, D₂O exchangeable), 7.59 (s, 1H), 6.00 - 5.45 (m, 3H), 4.70 (t, J= 8.5 Hz, 1H), 4.33 (dd, 7=21.4, 12.6 Hz, 1H), 4.15 - 3.88 (m, 1H), 2.75 (s, 3H), 2.67-2.57 (m, 1H), 2.382.07 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-J₆) δ-58.72,-176.42; MS (ES+): 595.1/597.1 (M+l); (ES-): 593.1/595.1 (M-l); Analysis calculated for C23Hi9BrF₄NsO2 1.75H2O. HCl.: C, 41.65; H, 3.57; Cl, 5.34; N, 16.89; Found: C, 41.92; H, 3.47; Cl, 5.35; N, 16.51.

Scheme 322

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-2-azabicyclo[3.1.0]hexane3-carboxamide (322c)

Step-1: Préparation of (lR,3S,5R)-terributyl 3-((6-bromo-5-fluoro-3-methylpyridin-2yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (322a)

Compound 322a was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1,0]hexane-3-carboxylic acid

557 (259a) (0.322 g, 1.415 mmol) in DCM (12 mL) and 6-bromo-5-fluoro-3-methylpyridin-2amine (0.290 g, 1.415 mmol; CAS # 2086189-45-7) to afford after purification as described in step-1 of scheme-317, (lR,3S,5R)-ter/-butyl 3-((6-bromo-5-fluoro-3-methylpyridin-2yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (322a) (0.463 g, 79% yield) as a 5 white solid; MS (ES+): 414.1/416.1(M+1).

Step-2: Préparation of (lR,3S,5R)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (322b)

Compound 322b was prepared according to the procedure reported in step-2 of scheme-1, from ( 1 R,3S,5R)-tert-butyl 3-((6-bromo-5-fluoro-3-methylpyridin-2-yI)carbamoyl)-210 azabicyclo[3.1.0]hexane-2-carboxylate (322a) (0.46 g, 1.110 mmol) in DCM (6 mL) using TFA (0.599 mL, 7.77 mmol) to afford after purification as described in step-2 of scheme-319 TFA sait of (lR,3S,5R)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (322b) (0.54 g, 100% yield); ’H NMR (300 MHz, DMSO-cfe) δ 10.81 (s, 1H), 9.88 (s, 1H), 9.21 (s, 1H), 7.93 (d, J= 8.4 Hz, 1H), 4.48 - 3.96 (m, 1H), 3.46-3.19 (m, 1H), 2.67 - 2.54 (m, 1H), 2.23-2.05 (m, 4H), 1.91-1.80 (m, 1H),

1.02 - 0.87 (m, 1H), 0.88 - 0.74 (m, 1H); ’⁹F NMR (282 MHz, DMSO-î/₆) δ -74.31, -117.92; MS (ES+): 314.0/316.0 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-220 azabicyclo[3.1.0]hexane-3-carboxamide (322c)

Compound 322c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of (lR,3S,5R)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (322b) (66.4 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-925 yl)acetic acid (184f) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-2-azabicycIo[3.1.0]hexane3-carboxamide (322c) (45 mg, 55% yield) HCl sait as a white solid; ’H NMR (300 MHz,

DMSO-Jé) δ 10.34 (s, 1H, D₂O exchangeable), 8.80 (s, 1H), 8.60 (s, 1H), 8.56 (s, 2H, D₂O exchangeable), 7.82 (d, J= 8.4 Hz, 1H), 7.59 (s, 1H), 5.94 (d, J= 18.0 Hz, 1H), 5.71 (d, J= 17.9 Hz, 1H), 4.37 (dd, J= 9.2, 5.5 Hz, 1H), 3.97 - 3.86 (m, 1H), 2.75 (s, 3H), 2.42 (dd, J = 13.4, 9.4 Hz, 1H), 2.33 - 2.17 (m, 1H), 2.02 (s, 3H), 2.00 - 1.88 (m, 1H), 1.21 - 1.05 (m, 1H), 0.79 - 0.68 (m, 1H); ^I9F NMR (282 MHz, DMSO) δ-58.70,-119.16. MS (ES+):

558

620.1/622.1 (M+l); (ES-): 618.0/620.0 (M-l); Analysis calculated for C26H22BrF4N7O2.H2O.HCI: C, 46.27; H, 3.73; Cl, 5.25; N, 14.53; Found: C, 46.1; H, 4.02; Cl, 5.16; N, 14.25.

Scheme 323

184f ^323a

Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromo-3-methylpyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2carboxamide (323a)

Compound 323a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyI-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (60 mg, 0.137 mmol) in DMF (1.5 mL) using HCl sait of (2S,4R)-N-(6bromo-3-methylpyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (209a) (48.3 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (2S,4R)-115 (2-(4-amino-8-methyl-6-(trifIuoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo3-methylpyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (323a) (45 mg, 53% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-dô) (a mixture of two rotamers) δ 10.83 and 10.50 (2s, 1H, D₂O exchangeable), 8.72 (s, 1H), 8.48 and 8.46 (2s, 1H), 8.14 (s, 2H, D₂O exchangeable), 7.73 and, 7.59 (2d, J= 8.0 Hz, 1H), 7.55 and 7.51 (2s, 1H), 7.41 (d,

J= 7.9 Hz, 1H), 5.74 (d, J= 18.0 Hz, 1H), 5.49 (d, J= 17.9 Hz, 1H), 4.62 (dd, J= 9.8, 7.5 Hz, 1H), 4.27 (dd, J= 18.1, 12.0 Hz, 1H), 3.89 (dd, J= 35.2, 12.0 Hz, 1H), 2.71 (s, 3H), 2.61 -2.54 (m, 1H), 2.21 -2.00 (m, 1H), 1.95 (s, 3H), 1.65 (d,7=21.1 Hz, 3H); ¹⁹FNMR(282 MHz, DMSO-î/₆) δ -58.59, -139.97. MS (ES+): 622.1/624.1 (M+l); (ES-): 620.1/622.1 (ΜΙ).; Analysis calculated for C26H24BrF4N7O2.L75H2O.HCl: C, 45.23; H, 4.16; Cl, 5.14; N,

14.20; Found: C, 45.34; H, 4.07; Cl, 4.99; N, 14.15

Scheme 324

Préparation of (S)-3-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)thiazolidine-2-carboxamide (324a)

Compound 324a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (60 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of (S)-N-(6bromopyridin-2-yl)thiazolidine-2-carboxamide (171a) (55.1 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (S)-3-(2-(4-amino-8-methyl-610 (trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)thiazolidine2-carboxamide (324a)(75 mg, 92% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-dô) (a mixture of two rotamers) δ 11.30and 11.08 (2s, 1H, D₂O exchangeable), 8.78 (s, 1H), 8.59 (s, 1H), 8.51 (s, 2H, D₂O exchangeable), 8.07and 7.97 (2d, J= 8.2 Hz, 1H), 7.82 and 7.74 (2t, J= 8.0 Hz, 1H), 7.59 (s, 1H), 7.42 and 7.35 (2d, J= 7.7 Hz, 1H), 5.86 - 5.54 (m, 3H), 4.42-4.28 (m, 1H), 4.18-4.05 (m, 1H), 3.44 - 3.31 (m, 2H), 2.75 (s, 3H); ¹⁹F

NMR (282 MHz, DMSO-î/₆) δ -58.74; MS (ES+): 594.1/596.1 (M+l); (ES-): 592.0/594.0 (M-l); Analysis calculated for C₂₃Hi₉BrF₃N₇O₂S.1.25H₂O.1.15HCl: C, 41.93; H, 3.47; Cl, 6.19; N, 14.88; Found: C, 41.81; H, 3.38; Cl, 5.92; N, 14.71.

Scheme 325

325c

Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloropyrazin-2-yl)-4-fluoropyrrolidine-2-carboxamide (325c) Step-1: Préparation of (2S,4R)-terZ-butyl 2-((6-chloropyrazin-2-yl)carbamoyl)-45 fluoropyrrolidine-l-carboxylate (325a)

Compound 325a was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4R)-l-(Zeri-butoxycarbonyl)-4-fluoropyrrolidine-2-carboxylic acid (321a) (0.466 g, 2 mmol) in DCM (15 mL) and 6-chloropyrazin-2-amine (0.259 g, 2.00 mmol; CAS # 3333228-4) to afford after purification as described in step-1 of scheme-317, (2S,4R)-ZerZ-butyl 210 ((6-chIoropyrazin-2-yl)carbamoyl)-4-fluoropyrrolidine-l-carboxylate (325a) (0.36 g, 52% yield) as a clear gel; MS (ES+): 345.1(M+1).

Step-2: Préparation of (2S,4R)-N-(6-chloropyrazin-2-yl)-4-fluoropyrrolidine-2-carboxamide (325b)

Compound 325b was prepared according to the procedure reported in step-2 of scheme-1, 75 from (2S,4R)-/er/-butyl 2-((6-chloropyrazin-2-yl)carbamoyl)-4-fluoropyrrolidine-lcarboxylate (325a) (0.35 g, 1.015 mmol) in DCM (5.5 mL) using TFA (0.547 mL, 7.11 mmol) to afford after purification as described in step-2 of scheme-319, HCl sait of (2S,4R)N-(6-chloropyrazin-2-yl)-4-fluoropyrrolidine-2-carboxamide (325b) (0.220 g, 77% yield); ’H NMR (300 MHz, DMSO-dô) δ 11.77 (s, 1H), 9.29 (s, 1H), 8.62 (s, 1H), 5.51 (d, J= 52.4 Hz, 20 1H), 4.64 (dd, J= 10.9, 7.3 Hz, 1H), 3.67 - 3.61 (m, 1H), 3.59 - 3.47 (m, 1H), 2.93 - 2.67

561 (m, 1H), 2.45 - 2.17 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-cfc) δ -175.30; MS (ES+): 245.0 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloropyrazin-2-yl)-4-fluoropyrrolidine-2-carboxamide (325c)

Compound 325c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (2S,4R)-N-(6-chIoropyrazin-2-yl)-4-fluoropyrrolidine-2-carboxamide (325b) (38.5 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(4-amino-8-methyl-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (2S,4R)-l-(2-(4-amino8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloropyrazin-2yl)-4-fluoropyrrolidine-2-carboxamide (325c) (69 mg, 91% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-cfc) (a mixture of two rotamers) δ 11.68 and 11.37 (2s, 1H, D₂O exchangeable), 9.38 and 9.21 (2s, 1H), 8.78 (s, 1H), 8.67 - 8.50 (m, 3H, 2H D₂O exchangeable), 8.48 (s, 1H), 7.60 (s, 1H), 5.93 - 5.46 (m, 3H), 4.70 (t, J= 8.5 Hz, 1H), 4.34 (dd,<7=21.4, 12.6Hz, 1H),4.O1 (dd,.7=38.1, 12.5 Hz, 1H), 2.75 (s, 3H), 2.64-2.55 (m, 1H), 2.35 - 2.06 (m, 1H); ^I9F NMR (282 MHz, DMSO-J₆) δ -58.74, -176.51. MS (ES+): 551.2 (M+l); (ES-): 549.1 (M-l); Analysis calculated for C₂3Hi9ClF₄N₈O₂ 1.75 H₂O. 0.85HC1.: C, 45.03; H, 3.84; Cl, 10.69; N, 18.27; Found: C, 45.06; H, 3.84; Cl, 10.59; N, 18.07.

Scheme 326

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyI)-N-(6-chloropyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (326c)

Step-1 : Préparation of (lR,3S,5R)-terZ-butyl 3-((6-chloropyrazin-2-yl)carbamoyl)-2azabicyclo [3.1.0]hexane-2-carboxylate (326a)

Compound 326a was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(terZ-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (259a) (0.455 g, 2 mmol) and 6-chloropyrazin-2-amine (0.259 g, 2 mmol; CAS # 33332-2810 4) to afford after purification as described in step-1 of scheme-317 (lR,3S,5R)-ZerZ-butyl 3((6-chloropyrazin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (326a) (0.245 g, 36% yield) as a clear gel; MS (ES+): 339.1 (M+l).

Step-2: Préparation of (lR,3S,5R)-N-(6-chloropyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (326b)

Compound 326b was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-ZerZ-butyl 3-((6-chloropyrazin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane2-carboxylate (326a) (0.24 g, 0.708 mmol) in DCM (4 mL) using TFA (0.382 mL, 4.96 mmol) to afford after purification as described in step-2 of scheme-319, (lR,3S,5R)-N-(6chloropyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (326b) (0.14 g, 72% yield)

HCl sait; ’H NMR (300 MHz, DMSO-î/₆) δ 11.66 (s, 1H), 9.26 (s, 1H), 8.61 (s, 1H), 4.21 (dd, J= 10.9, 7.8 Hz, 1H), 3.32 - 3.26 (m, 1H), 2.62 (dd, J= 12.8, 7.8 Hz, 1H), 2.22 - 1.99 (m, 1H), 1.92- 1.68 (m, 1H), 1.00-0.58 (m, 2H); MS (ES+): 239.0 (M+l).

563

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloropyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (326c)

Compound 326c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (lR,3S,5R)-N-(6-chloropyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (326b) (37.7 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(4-amino8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(210 (4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6chIoropyrazin-2-yl)-2-azabicycIo[3.1.0]hexane-3-carboxamide (326c) (67 mg, 90 % yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-<7₆) δ 11.19 (s, 1H, D₂O exchangeable), 9.23 (s, 1H), 8.79 (s, 1H), 8.63 (s, 3H, 2HD₂O exchangeable), 8.48 (s, 1H), 7.62 (s, 1H), 5.99 (d, J= 18.0 Hz, 1H), 5.71 (d, J= 18.0 Hz, 1H), 4.46 (dd, J= 9.1, 5.7 Hz, 1H), 4.03 - 3.80 (m,

1H), 2.78 (s, 3H), 2.38 (dd, J= 13.5, 9.2 Hz, 1H), 2.32 - 2.18 (m, 1H), 2.02 - 1.87 (m, 1H),

1.21 - 1.01 (m, 1H), 0.78 - 0.67 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-t/₆) δ -58.76; MS (ES+): 545.2 (M+l); (ES-): 543.1 (M-l); Analysis calculated for C₂₄H₂₀ClF₃N₈O₂.2H₂O. 0.75HC1.: C, 47.39; H, 4.10; Cl, 10.20; N, 18.42; Found: C, 47.57; H, 3.92; Cl, 10.02; N, 18.31.

Scheme 327

564

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifIuoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyI)-N-(6-chloropyrazin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3carboxamide (327c)

Step-1: Préparation of (lR,3S,5R)-ter/-butyl 3-((6-chloropyrazin-2-yl)carbamoyl)-5-methyl2-azabicyclo [3.1.0]hexane-2-carboxylate (327a)

Compound 327a was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(teri-butoxycarbonyl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (261a) (0.483 g, 2 mmol) and 6-chloropyrazin-2-amine (0.259 g, 2 mmol) to afford after purification as described in step-1 of scheme-317, (lR,3S,5R)-tert-butyl 3-((6-chloropyrazin2-yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (327a) (0.091 g, 13% yield) as a clear gel; MS (ES+): 353.1 (M+l).

Step-2: Préparation of (lR,3S,5R)-N-(6-chloropyrazin-2-yl)-5-methyl-2azabicyclo[3.1,0]hexane-3-carboxamide (327b)

Compound 327b was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-/ert-butyl 3-((6-chloropyrazin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexane-2-carboxylate (327a)(0.085 g, 0.241 mmol) in DCM (1.5 mL) using TFA (0.130 mL, 1.686 mmol) to afford after purification as described in step-2 of scheme319, (lR,3S,5R)-N-(6-chloropyrazin-2-yI)-5-methyI-2-azabicyclo[3.1.0]hexane-3carboxamide (327b) (0.055 g, 79% yield) HCl sait; Ή NMR (300 MHz, DMSO-îZ₆) δ 11.68 (s, 1H), 9.26 (s, 1H), 8.61 (s, 1H), 4.24 (dd, J= 11.1, 7.7 Hz, 1H), 3.10 (dd, J= 6.9, 2.4 Hz, 1H), 2.74-2.60 (m, 1H), 2.00 (t, J= 11.8 Hz, 1H), 1.26 (s, 3H), 1.03 (dd, J= 7.2, 2.5 Hz, 1H), 0.79 (t, J= 7.0 Hz, 1H); MS (ES+): 253.1 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloropyrazin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3 -carboxamide (327c)

Compound 327c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (lR,3S,5R)-N-(6-chloropyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane3-carboxamide (327b) (39.6 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(4amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (1841) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-chloropyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (327c) (64 mg, 84% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 11.18

565 (s, 1H, D₂0 exchangeable), 9.23 (s, 1H), 8.78 (s, 1H), 8.60 (s, 1H), 8.53 (s, 2H, D₂O exchangeable), 8.48 (s, 1H), 7.60 (s, 1H), 5.93 (d, J= 18.0 Hz, 1H), 5.66 (d, J= 17.9 Hz, 1H), 4.41 (dd, J= 9.1, 6.1 Hz, 1H), 3.73 (dd, J= 5.5, 2.3 Hz, 1H), 2.76 (s, 3H), 2.56 - 2.52 (m, 1H), 2.03 (dd, J= 13.3, 6.0 Hz, 1H), 1.32 (s, 3H), 1.08 - 0.99 (m, 1H), 0.92 - 0.83 (m,

1H); ^{10 * * * * 15 * * * 19 20}F NMR (282 MHz, DMSO-<7₆) δ -58.73; MS (ES+): 559.1 (M+l); (ES-): 557.1 (M-l);

Analysis calculated for C25H22CIF3N8O2 2.25H₂O. 0.75HCL: C, 47.90; H, 4.38; Cl, 9.90; N, 17.88; Found: C, 47.92; H, 4.18; Cl, 9.91; N, 17.77.

Scheme 328

Me

NC^CN

Cul, K2CO3

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methoxy-6-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (328f)

Step-1: Préparation of 2-amino-6-methoxy-5-methyl-lH-indole-3-carbonitrile (328b)

Compound 328b was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-5-methoxy-4-methylphenyl)-2,2,2-trifluoroacetamide (328a) (7.4 g, 23.71 mmol; CAS # 1262881-27-5) in DMSO (30 mL) using malononitrile (1.792 mL, 28.5 mmol),

L-proline (0.546 g, 4.74 mmol), Cul (452 mg, 2.371 mmol), a solution of K2CO3 (6.55 g,

47.4 mmol) in water (30 mL) and heating at 60 °C for 16 h under an argon atmosphère. This gave after workup and purification [S1O2 gel (80 g), EtOAc/methanol (9:1) in hexane from 0 50%] 2-amino-6-methoxy-5-methyl-lH-indole-3-carbonitrile (328b) (1.95 g, 41% yield) as a grey solid; ^[H NMR (300 MHz, DMSO-ife) δ 10.43 (s, 1H), 6.89 (s, 1H), 6.75 (s, 1H), 6.49 (s, 2H), 3.73 (s, 3H), 2.21 -2.07 (m, 3H); MS (ES+): 202.1 (M+l); (ES-): 200.1 (M-l).

Step-2: Préparation of 7-methoxy-6-methyI-9H-pyrimido[4,5-b]indol-4-amine (328c) Compound 328c was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-6-methoxy-5-methyl-lH-indole-3-carbonitrile (328b) (1.9 g, 9.44 mmol) in

566 éthanol (60 mL) using formamidine acetate (7.94 g, 76 mmol) to afford after workup AcOH sait of 7-methoxy-6-methyl-9H-pyrimido[4,5-b]indol-4-amine (328c) (2.72 g, 81% yield) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-J₆) δ 8.53 (s, 1H), 8.45 (s, 1H), 8.32 (s, 1H), 7.46 (s, 1H), 5.29 (s, 2H), 3.92 (s, 3H), 2.32 (s, 3H); MS (ES+): 229.1 (M+l).

Step-3: Préparation of tert-butyl 2-(4-amino-7-methoxy-6-methyl-9H-pyrimido[4,5-b]indol9-yl)acetate (328d)

To a mixture of 7-methoxy-6-methyl-9H-pyrimido[4,5-b]indol-4-amine (328c) AcOH sait (2 g, 6.94 mmol) in DMF (50 mL) was added césium carbonate (6.78 g, 20.81 mmol) at 0 °C under N2 and stirred at the same température for 20 min. To this mixture was added tertio butyl 2-bromoacetate (1.128 mL, 7.63 mmol) and allowed to RT over a period of 16 h at RT. The reaction mixture was quenched by adding water. The solid separated was filtered and purified by reverse phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0 -100%] to afford tert-butyl 2-(4-amino-7-methoxy-6methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (328d) (0.2g, 8% yield) as a pale yellow solid;

Ή NMR (300 MHz, DMSO-J₆) δ 8.24 (s, 1H), 8.15 (s, 1H), 7.19 (s, 3H), 5.12 (s, 2H), 3.88 (s, 3H), 2.29 (s, 3H), 1.41 (s, 9H); MS (ES+): 343.2 (M+l).

Step-4: Préparation of 2-(4-amino-7-methoxy-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (328e)

Compound 328e was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-7-methoxy-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (328d) (0.19 g, 0.555 mmol) using 20% TFA in DCM (3.18 mL, 8.32 mmol) to afford the TFA sait of 2-(4-amino-7-methoxy-6-methyl-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (328e) (285 mg) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-J₆) δ 8.53 (s, 1H), 8.45 (s, 1H), 8.32 (s, 1H), 7.46 (s, 1H), 5.29 (s, 2H), 3.92 (s, 3H), 2.32 (s, 3H); MS (ES+): 287.10 (M+l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methoxy-6-methyl-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (3281)

Compound 328f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-methoxy-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (328e) (81 mg, 0.157 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (50mg, 0.157 mmol), HATU (90 mg, 0.235 mmol), DIPEA (0.137 mL, 0.785 mmol) and stirring at RT for 16 h.

567

This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(2-(4amino-7-methoxy-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)2-azabicyclo[3.1.0]hexane-3-carboxamide (328f) (51 mg, 59% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/ô) δ 10.73 (s, 1H, D₂O exchangeable), 8.54 (s, 1H), 8.43 (s, 2H, D₂O exchangeable), 8.31 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.72 (t, J= 8.0 Hz, 1H),

7.33 (d, J= 7.7 Hz, 1H), 7.25 (s, 1H), 5.71 (d, J= 17.2 Hz, 1H), 5.46 (d, J= 17.2 Hz, 1H), 4.41 (dd, J= 9.1, 5.6 Hz, 1H), 4.00 - 3.86 (m, 4H), 2.46 - 2.12 (m, 5H), 2.00 - 1.88 (m, 1H), 1.12-1.01 (m, 1H), 0.79-0.65 (m, 1H). MS (ES+): 550.1/552.1 (M+l); (ES-): 548.1/550.1 (M-l); Analysis calculated for C₂₅H₂4BrN₇O3 2H₂O. HCL: C, 48.20; H, 4.69; Cl, 5.69; N,

15.74; Found: C, 48.19; H, 4.35; Cl, 5.70; N, 15.69.

Scheme 329

328e ^329a

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methoxy-6-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyI)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide 15 (329a)

Compound 329a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-methoxy-6-methyl-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (328e) (81 mg, 0.157 mmol) in DMF (2 mL) using HCI sait of (lR,3S,5R)-N-(6bromopyridin-2-yI)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (52.2 mg, 0.157 20 mmol), HATU (90 mg, 0.236 mmol), DIPEA (0.137 mL, 0.785 mmol) and stirring at RT for

h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(2(4-amino-7-methoxy-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (329a) (56 mg, 63% yield) HCl sait as a white solid; ’HNMR (300 MHz, DMSO-î/ô) δ 10.72 (s, 1H, D₂O exchangeable), 8.53 (s, 25 1H), 8.40 (s, 2H, D₂O exchangeable), 8.30 (s, 1H), 8.02 (d, J= 8.2 Hz, 1H), 7.72 (t, J= 8.0

Hz, 1H), 7.33 (d, J= 7.7 Hz, 1H), 7.25 (s, 1H), 5.67 (d, J= 17.3 Hz, 1H), 5.40 (d, J= 17.2

568

Hz, 1H), 4.37 (dd, J= 9.1, 6.1 Hz, 1H), 3.92 (s, 3H), 3.70 (dd, J= 5.6, 2.3 Hz, 1H), 2.50 2.43 (m, 1H), 2.30 (s, 3H), 2.00 (dd, J= 13.2, 6.0 Hz, 1H), 1.32 (s, 3H), 1.10 - 0.97 (m, 1H), 0.97 - 0.82 (m, 1H); MS (ES+): 564.1/566.1 (M+l); (ES-): 562.1/564.0 (M-l); Analysis calculated for C₂₆H26BrN₇O3 1.75H₂O. 1.15HC1.: C, 48.95; H, 4.84; Cl, 6.39; N, 15.37;

Found: C, 48.93; H, 4.87; Cl, 6.37; N, 15.47.

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-fluoro-6-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (330f)

Step-1: Préparation of 2-amino-6-fluoro-5-methoxy-lH-indole-3-carbonitrile (330b) Compound 330b was prepared according to the procedure reported in step-1 of scheme-11, fromN-(2-bromo-5-fluoro-4-methoxyphenyl)-2,2,2-trifluoroacetamide (330a) (3.5 g, 11.07 mmol; CAS # 1262881-29-7) in DMSO (15mL) using malononitrile (0.837 mL, 13.29 mmol), L-proline (0.255 g, 2.215 mmol), Cul (211 mg, 1.107 mmol), a solution ofKùCCE (3.06 g, 22.15 mmol) in water (15 mL) to afford after workup and purification as described in step-1 of scheme-328, 2-amino-6-fluoro-5-methoxy-lH-indole-3-carbonitrile (330b)(0.95 g, 42% yield) as a yellow solid; MS (ES+): 206.1 (M+l); (ES-): 204.0 (M-l).

Step-2: Préparation of 7-fluoro-6-methoxy-9H-pyrimido[4,5-b]indol-4-amine (330c) Compound 330c was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-6-fluoro-5-methoxy-lH-indole-3-carbonitrile (330b) (0.9 g, 4.39 mmol) in éthanol (30 mL) using formamidine acetate (3.69 g, 35.1 mmol) to afford after workup AcOH sait of 7-fluoro-6-methoxy-9H-pyrimido[4,5-b]indol-4-amine (330c) (0.98 g, 85% yield) as a

569 pale-yellow solid; Ή NMR (300 MHz, DMSO-J₆) δ 11.74 (s, 1H), 8.20 (s, 1H), 8.04 (d, J= 8.4 Hz, 1H), 7.35 - 7.16 (m, 3H), 3.93 (s, 3H); MS (ES+): 233.1 (M+l).

Step-3: Préparation of tert-butyl 2-(4-amino-7-fluoro-6-methoxy-9H-pyrimido[4,5-b]indol9-yl)acetate (330d)

Compound 330d was prepared according to the procedure reported in step-1 of scheme-1/ step-3 of scheme-328, from AcOH sait of 7-fhioro-6-methoxy-9H-pyrimido[4,5-b]indol-4amine (330c) (950 mg, 3.62 mmol) in DMF (25 mL) using tert-butyl 2-bromoacetate (0.589 mL, 3.98 mmol) and CS2CO3 (2.95 g, 9.06 mmol) to afford after workup and purification as described in step-3 of scheme-328, tert-butyl 2-(4-amino-7-fluoro-6-methoxy-9H10 pyrimido[4,5-b]indol-9-yl)acetate (330d) (0.89 g, 71% yield) as a pale yellow solid; MS (ES+): 347.2 (M+l).

Step-4: Préparation of 2-(4-amino-7-fluoro-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (330e)

Compound 330e was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-7-fluoro-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (330d) (0.87 g, 2.51 mmol) using 20% TFA in DCM (14.42 mL, 37.7 mmol) to afford after workup TFA sait of 2-(4-amino-7-fluoro-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (330e) (1.3 g) as a pale-yellow solid; ‘H NMR (300 MHz, DMSO-tZ₆) δ 8.73 (s, 2H), 8.58 (s, 1H), 8.24 (d, J= 8.2 Hz, 1H), 7.91 (d, J= 11.7 Hz, 1H), 5.24 (s, 2H), 3.98 (s, 3H); MS (ES+):

291.1 (M+l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-7-fluoro-6-methoxy-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (330f)

Compound 330f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-fluoro-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (330e) (81 mg, 0.157 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (50mg, 0.157 mmol), HATU (90 mg, 0.235 mmol), DIPEA (0.137 mL, 0.785 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(2-(430 amino-7-fluoro-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)2-azabicyclo[3.1.0]hexane-3-carboxamide (330f) (56 mg, 64.4 % yield) HCl sait as a white solid; ‘H NMR (300 MHz, DMSO-ife) δ 10.77 (s, 1H, D₂O exchangeable), 8.54 (s, 3H, 2H D₂O exchangeable), 8.24 (d, J= 8.2 Hz, 1H), 8.01 (d, J= 8.1 Hz, 1H), 7.77 - 7.63 (m, 2H), 7.32 (d, J= 7.7 Hz, 1H), 5.71 (d, J= 17.4 Hz, 1H), 5.36 (d, J= 17.3 Hz, 1H), 4.42 (dd, J=

570

9.0, 5.4 Hz, 1H), 3.97 (s, 3H), 3.93 - 3.81 (m, 1H), 2.39 - 2.27 (m, 1H), 2.27 - 2.14 (m, 1H), 2.00 - 1.82 (m, 1H), 1.19 - 0.96 (m, 1H), 0.84 - 0.75 (m, 1H). ^I9F NMR (282 MHz, DMSOdd) δ -133.52. MS (ES+): 554.1/556.1 (M+l); (ES-): 552.1/554.1 (M-l); Analysis calculated for C₂₄H₂iBrFN₇O₃ 2H₂O. 0.9HC1.: C, 46.25; H, 4.19; Cl, 5.12; N, 15.73; Found: C, 46.18;

H, 3.92; Cl, 5.08; N, 15.62.

Scheme 331

Préparation· of (lR,3S,5R)-2-(2-(4-amino-7-fluoro-6-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (331a)

Compound 331a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-fIuoro-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (330e) (78 mg, 0.150 mmol) in DMF (2 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (50 mg, 0.150 mmol), HATU (86 mg, 0.225 mmol), DIPEA (0.131 mL, 0.752 mmol) and stirring at RT for 16 h. This gave after workup and purification using as described in scheme-303, (1R,3S,5R)2-(2-(4-amino-7-fluoro-6-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (331a) (48mg, 56% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSOX) δ 10.77 (s, 1H, D₂O exchangeable), 8.64 - 8.28 (m, 3H, 2H D₂O exchangeable), 8.21 (d, J= 8.2 Hz, 1H), 8.02 (d, J= 8.1 Hz, 1H), 7.80 - 7.58 (m, 2H), 7.32 (d, J= 7.7 Hz, 1H), 5.66 (d, J= 17.4 Hz, 1H), 5.30 (d, J= 17.3 Hz, 1H), 4.37 (dd, J= 9.1, 5.9 Hz, 1H), 3.96 (s, 3H), 3.70 - 3.64 (m, 1H), 2.48 2.41 (m, 1H), 1.98 (dd, J= 13.3, 5.8 Hz, 1H), 1.31 (s, 3H), 1.06-0.90 (m, 2H); ¹⁹FNMR (282 MHz, DMSO-Jô) δ -133.30. MS (ES+): 568.1/570.1 (M+l); (ES-): 566.1/568.1 (M-l);

Analysis calculated for C₂₅H₂₃BrFN₇O₃ 2H₂O. 1.1HC1.: C, 46.59; H, 4.39; Cl, 6.05; N, 15.21; Found: C, 46.52; H, 4.33; Cl, 6.12; N, 15.15.

Scheme 332

571

332c

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloro-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane3-carboxamide (332c)

Step-1 : Préparation of (1 R,3S,5R)-ZerZ-butyl 3-((6-chloro-3-methylpyridin-2-yl)carbamoyl)5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (332a)

Compound 332a was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(terZ-butoxycarbonyl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (261a) (0.35 g, 1.451 mmol) and 6-chloro-3-methylpyridin-2-amine (0.207 g, 1.451 mmol) to afford after purification as described in step-1 of scheme-317, (lR,3S,5R)-fërt-butyl 3-((6-chloro-3-methylpyridin-2-yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexane-2carboxylate (332a) (0.196 g, 37% yield) as a white solid; MS (ES+): 366.2 (M+l). Step-2: Préparation of (lR,3S,5R)-N-(6-chloro-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (332b)

Compound 332b was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-terributyl 3-((6-chloro-3-methylpyridin-2-yl)carbamoyl)-5-methyI-2azabicyclo[3.1.0]hexane-2-carboxylate (332a) (0.19 g, 0.519 mmol) in DCM (2.8 mL) using TFA (0.280 mL, 3.64 mmol) to afford after purification as described in step-2 of scheme-319, (lR,3S,5R)-N-(6-chloro-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-320 carboxamide (332b) (137 mg, 99% yield); *H NMR (300 MHz, DMSO-dô) δ 10.89 (s, 1H), 10.12 (s, 1H), 9.19 (d, J= 10.0 Hz, 1H), 7.82 (d, J= 8.0 Hz, 1H), 7.41 (d, J= 8.0 Hz, 1H), 4.36-4.22 (m, 1H), 3.11 - 3.04 (m, 1H), 2.65 (dd, J= 12.6, 7.6 Hz, 1H), 2.16 (s, 3H), 2.05 21436

572

1.95 (m, 1H), 1.27 (s, 3H), 1.13 - 1.03 (m, 1H), 0.84 - 0.72 (m, 1H); MS (ES+): 266.1 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloro-3-methylpyridin-2-yl)₇5-methyl-25 azabicyclo [3.1.0]hexane-3 -carboxamide (332c)

Compound 332c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (lR,3S,5R)-N-(6-chloro-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (332b) (41.4 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-910 yl)acetic acid (184f) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloro-3-methylpyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane3-carboxamide (332c) (55 mg, 70% yield) HCl sait as a white solid; *H NMR (300 MHz,

DMSO-î/ô) δ 10.31 (s, 1H, D₂O exchangeable), 8.79 (s, 1H), 8.59 (s, 1H), 8.53 (s, 2H, D₂O exchangeable), 7.72 (d, J= 8.0 Hz, 1H), 7.58 (s, 1H), 7.31 (d, J= 7.9 Hz, 1H), 5.90 (d, J= 18.0 Hz, 1H), 5.66 (d, J= 17.9 Hz, 1H), 4.38 (dd, 9.1, 5.6 Hz, 1H), 3.69 (dd, J= 5.6, 2.4 Hz, 1H), 2.74 (s, 3H), 2.62-2.56 (m, 1H), 2.10 - 1.94 (m, 4H), 1.33 (s, 3H), 1.12-0.98 (m, 1H), 0.96 - 0.78 (m, 1H). ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -58.70; MS (ES+): 572.2 (M+l);

(ES-): 570.1 (M-l); Analysis calculated for C₂7H₂5C1F₃N7O₂ 2.25H₂O.0.75HCl: C, 50.68; H,

4.77; Cl, 9.70; N, 15.32; Found: C, 50.55; H, 4.68; Cl, 9.70; N, 15.18.

Scheme 333

333c

573

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (333c)

Step-1: Préparation of (!R,3S,5R)-ZerZ-butyl 3-((6-bromo-5-fluoro-3-methylpyridin-25 yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (333a)

Compound 333a was prepared according to the procedure reported in step-1 of scheme-53, from (lR,3S,5R)-2-(ZerZ-butoxycarbonyl)-5-methyl-2-azabicyclo[3.1,0]hexane-3-carboxylic acid (261a) (0.35 g, 1.451 mmol) and 6-bromo-5-fluoro-3-methylpyridin-2-amine (0.297 g, 1.451 mmol) to afford after purification as described in step-1 of scheme-317, (1R,3S,5R)10 ZerZ-butyl 3-((6-bromo-5-fluoro-3-methylpyridin-2-yl)carbamoyl)-5-methyl-2azabicyclo[3.1.0]hexane-2-carboxylate (333a) (0.560 g, 90% yield) as a white solid; MS (ES+): 428.1/430.1(M+1).

Step-2: Préparation of (lR,3S,5R)-N-(6-bromo-5-fhioro-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (333b)

Compound 333b was prepared according to the procedure reported in step-2 of scheme-1, from (lR,3S,5R)-ZerZ-butyl 3-((6-bromo-5-fluoro-3-methylpyridin-2-yl)carbamoyl)-5-methyl2-azabicyclo[3.1.0]hexane-2-carboxylate (333a) (0.550 g, 1.284 mmol) in DCM (7 mL) using TFA (0.693 mL, 8.99 mmol) to afford after purification as described in step-2 of scheme-319, (lR,3S,5R)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-5-methyl-220 azabicyclo[3.1.0]hexane-3-carboxamide (333b) (408 mg, 97% yield); *H NMR (300 MHz, DMSO-dô) δ 10.92 (s, 1H), 10.08 (s, 1H), 9.27 - 9.11 (m, 1H), 7.93 (d, J= 8.4 Hz, 1H), 4.38 -4.21 (m, 1H), 3.07 (d, J= 6.6 Hz, 1H), 2.65 (dd, J= 12.6, 7.6 Hz, 1H), 2.17 (s, 3H), 1.99 (t, J= 11.8 Hz, 1H), 1.27 (s, 3H), 1.15 - 1.01 (m, 1H), 0.81-0.71 (m, 1H); MS (ES+):

328.1/330.1(M+1).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (333c)

Compound 333c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (lR,3S,5R)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-5-methyl-230 azabicyclo[3.1.0]hexane-3-carboxamide (333b) (49.9 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (184f) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,521436

574 . b]indol-9-yl)acetyI)-N-(6-bromo-5-fluoro-3-methyIpyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (333c) (68 mg, 78 % yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-J₆) δ 10.36 (s, 1H, D₂O exchangeable), 8.80 (s, 1H), 8.74 - 8.44 (m, 3H, 2H D₂O exchangeable), 7.82 (d, 8.5 Hz, 1H), 7.59 (s, 1H), 5.90 (d, J= 18.0 Hz,

1H), 5.66 (d, J= 17.9 Hz, 1H), 4.35 (dd, J= 9.2, 5.7 Hz, 1H), 3.69 (dd, J= 5.6, 2.4 Hz, 1H),

2.74 (s, 3H), 2.62 - 2.54 (m, 1H), 2.08 - 1.97 (m, 4H), 1.33 (s, 3H), 1.09 - 0.96 (m, 1H), 0.94 - 0.86 (m, 1H). ¹⁹F NMR (282 MHz, DMS0-î/₆) δ -58.72, -119.20. MS (ES+): 634.1/636.1 (M+l); (ES-): 632.1/634.1 (M-l); Analysis calculated for C₂₇H₂4BrF4N₇O₂ H₂O.HC1: C, 47.07; H, 3.95; Cl, 5.15; N, 14.23; Found: C, 47.00; H, 3.75; Cl, 4.92; N, 14.06.

Scheme 334

H

ISL ^Br

O 8a

HATU, DIPEA

274f

334a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (334a)

Compound 334a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (274f) (75 mg, 0.23 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (92 mg, 0.276 mmol), HATU (175 mg, 0.460 mmol), DIPEA (0.160 mL, 0.920 mmol) and stirring at RT for 20 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6(trifluoromethoxy)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl2-azabicyclo[3.1.0]hexane-3-carboxamide (334a) (109 mg, 78% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.76 (s, 1H, D₂O exchangeable), 8.64 - 8.59 (m, 1H), 8.56 (s, 1H), 8.44 (s, 2H, D₂O exchangeable), 8.01 (d, J= 8.2 Hz, 1H), 7.76 (d, J= 9.0

Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.57 - 7.48 (m, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.72 (d, J= 17.4 Hz, 1H), 5.38 (d, J= 17.3 Hz, 1H), 4.36 (dd, J= 9.0, 6.0 Hz, 1H), 3.71 - 3.64 (m, 1H),

575

2.56 - 2.41 (m, 1H), 1.98 (dd, J= 13.3, 5.9 Hz, 1H), 1.30 (s, 3H), 1.05 - 0.97 (m, 1H), 0.97 0.90 (m, 1H); ^,9F NMR (282 MHz, DMSO-î/₆) δ -57.00; MS (ES+): 604.10 & 606.10 (M+l); MS (ES-): 602.00 & 604.00 (M-l); Analysis calculated for C25H2iBrF₃N₇O₃.HC1.1.25H₂O: C, 45.26; H, 3.72; N, 14.78; Cl, 5.34; Found: C, 45.13; H, 3.83; N, 14.66; Cl, 5.08.

Scheme 335

Préparation of ( 1 R,3S,5R)-2-(2-(4-amino-6-(4-fluorophenyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (335c) Step-1: Préparation of ethyl 2-(4-amino-6-(4-fluorophenyl)-9H-pyrimido[4,5-b]indoI-910 yl)acetate (335a)

Compound 335a was prepared and purified according to the procedure reported in step-1 of scheme-62, from ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (200 mg, 0.573 mmol) in dioxane (18 mL) using (4-fluorophenyl)boronic acid (120 mg, 0.859 mmol), césium carbonate (280 mg, 0.859 mmol) in water (2.2 mL), Pd(PPh₃)₂C12 (80 mg,

0.115 mmol) and heating at 100 °C for 18 h to give a mixture of ethyl 2-(4-amino-6-(4fluorophenyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (335a) and 2-(4-amino-6-(4fluorophenyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (335b) which was used as such for the next step; MS (ES+): 337.10 & 365.10 (M+l).

Step-2: Préparation of 2-(4-amino-6-(4-fluorophenyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic 20 acid (335b)

Compound 335b was prepared according to the procedure reported in step-4 of scheme-17, from mixture of ethyl 2-(4-amino-6-(4-fluorophenyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate

576 (335a) and 2-(4-amino-6-(4-fluorophenyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (335b) obtained in step-1 of this scheme (0.573 mmol) in THF (5 mL) and methanol (5 mL) using a solution of lithium hydroxide hydrate (147 mg, 3.44 mmol) in water (5 mL) and stirring at RT for 12 h. The reaction mixture was concentrated to dryness diluted with water (12 mL), 5 acidified with 4 N HCl to pH about 6 followed by filtration, washing with water, and drying under vacuum to afford a gray solid of 2-(4-amino-6-(4-fluorophenyl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (335b) (120 mg) which was used as such for the next step; MS (ES+): 337.10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(4-fluorophenyl)-9H-pyrimido[4,510 b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (335c)

Compound 335c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(4-fluorophenyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (335b) (60 mg, 0.178 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-215 azabicyclo[3.1.0]hexane-3-carboxamide (4a) (68.2 mg, 0.214 mmol), HATU (136 mg, 0.357 mmol), DIPEA (0.124 mL, 0.714 mmol) and stirring at RT for 20 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6-(4-fluorophenyl)9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (335c) (52 mg, 49% yield) HCl sait as a white solid; *H NMR (300 MHz,

DMSO-dô) δ 10.77 (s, 1H, D2O exchangeable), 8.78 (d, 1H), 8.67 (s, 3H, D2O exchangeable), 8.62 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.92 - 7.83 (m, 3H), 7.77 (d, J= 8.7 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.40 - 7.28 (m, 3H), 5.79 (d, J= 17.4 Hz, 1H), 5.45 (d, J= 17.3 Hz, 1H), 4.42 (dd, J =9.0, 5.6 Hz, 1H), 4.05-3.81 (m, 1H), 2.44-2.14 (m, 2H), 2.02- 1.84 (m, 1H), 1.17 -1.01 (m, 1H), 0.85-0.69 (m, 1H); ^I9FNMR(282 MHz, DMSO-dô) δ-116.03; MS (ES+):

600.10 & 602.15 (M+l); MS (ES-): 598.10 & 600.05 (M-l); Analysis calculated for

C₂9H23BrFN7O2.1.15HCL1.75H₂O: C, 51.69; H, 4.14; N, 14.55; Cl, 6.05; Found: C, 51.71; H, 4.14; N, 14.42; Cl, 5.91. .

Scheme 336

577

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(furan-2-yl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicycIo[3.1.0]hexane-3-carboxamide (336c)

Step-1: Préparation of ethyi 2-(4-amino-6-(furan-2-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (336a) .

Compound 336a was prepared and purified according to the procedure reported in step-1 of scheme-62, from ethyi 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (200 mg, 0.573 mmol) in dioxane (18 mL) using furan-2-ylboronic acid (96 mg, 0.859 mmol), césium carbonate (280 mg, 0.859 mmol) in water (2.2 mL), Pd(PPh3)₂Cl₂ (80 mg, 0.115 mmol) to give a mixture of ethyi 2-(4-amino-6-(furan-2-yl)-9H-pyrimido[4,5-b]indol-9yl)acetate (336a) and 2-(4-amino-6-(furan-2-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (336b) and this was used as such for the next step; MS (ES+): 309.10 & 337.20.

Step-2: Préparation of 2-(4-amino-6-(furan-2-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (336b)

Compound 336b was prepared according to the procedure reported in step-4 of scheme-17, from a mixture of ethyi 2-(4-amino-6-(furan-2-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (336a) and 2-(4-amino-6-(furan-2-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (336b) from step-1 above (0.573 mmol) in THF (5 mL) and methanol (5 mL) using a solution of lithium hydroxide hydrate (147 mg, 3.44 mmol) in water (5 mL) and stirring at RT for 23 h. This gave after workup a dark brown solid (190 mg) which was used as such for the next step; MS (ES+): 309.10 (M+l)

578

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(furan-2-yl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (336c)

Compound 336c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(furan-2-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (336b) (90 mg, 0.292 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (97 mg, 0.292 mmol), HATU (222 mg, 0.584 mmol), DIPEA (0.203 mL, 1.168 mmol) and stirring at RT for 20 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-610 (fiiran-2-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (336c) (29 mg, 17% yield foro three steps) HCl sait as a light brown solid. 'H NMR (300 MHz, DMSO-Jô) δ 10.77 (s, 1H, D2O exchangeable), 8.83 (d, J= 1.6 Hz, 1H), 8.67 (s, 2H, D₂0 exchangeable), 8.60 (s, 1H), 8.01 (d, 7= 8.2 Hz, 1H), 7.90 (dd, 7= 8.6, 1.5 Hz, 1H), 7,80 - 7.78 (m, 1H), 7.73 (d, 7= 8.5 Hz, 1H), 7.68 (d, 7 =

8.0 Hz, 1H), 7.31 (d, 7= 7.7 Hz, 1H), 7.04 (d, 7= 3.4 Hz, 1H), 6.65 (dd, 7= 3.4, 1.8 Hz, 1H),

5.72 (d, J= 17.3 Hz, 1H), 5.37 (d, 7= 17.3 Hz, 1H), 4.37 (dd, 7= 9.2, 5.8 Hz, 1H), 3.69 (dd, 7= 5.6, 2.3 Hz, 1H), 2.60-2.38 (m, 1H), 1.98 (dd, 7= 13.4, 5.9 Hz, 1H), 1.31 (s, 3H), 1.060.97 (m, 1H), 0.97 - 0.87 (m, 1H); MS (ES+): 586.10 & 588.20 (M+l); MS (ES-): 584.10 & 586.05 (M-l); Analysis calculated for C₂8H₂4BrN7O3.L0HC1.2.25H₂O: C, 50.69; H, 4.48; N,

14.78; Cl, 5.34; Found: C, 50.60; H, 4.34; N, 14.64; Cl, 5.09.

Scheme 337

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(4-fluorophenyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide 25 (337a)

Compound 337a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(4-fluorophenyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (335b) (60

579 mg, 0.178 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (71.2 mg, 0.214 mmol), HATU (136 mg, 0.357 mmol), DIPEA (0.124 mL, 0.714 mmol) and stirring at RT for 20 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6-(45 fluorophenyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (337a) (48 mg, 44% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-J₆) δ 10.78 (s, 1H, D₂O exchangeable), 8.77 (d, J= 1.7 Hz, 1H), 8.66 (s, 3H, D₂O exchangeable), 8.61 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.92 - 7.82 (m, 3H), 7.76 (d, J= 8.7 Hz, 1H), 7.69 (t, J= 7.9 Hz, 1H), 7.39 - 7.29 (m, 3H), 5.75 (d, J= 17.3 Hz,

1H), 5.40 (d, J= 17.3 Hz, 1H), 4.37 (dd, J= 9.1, 6.0 Hz, 1H), 3.71 (dd, J= 5.6, 2.4 Hz, 1H),

2.63-2.37 (m, 1H), 1.99 (dd, J= 13.1, 5.9 Hz, 1H), 1.31 (s, 3H), 1.07-0.98 (m, 1H), 0.980.89 (m, 1H).; ^I9FNMR(282 MHz, DMSO-î/₆) δ -116.03; MS (ES+): 614.10 & 616.15 (M+l); MS (ES-): 612.10 & 614.10 (M-l); Analysis calculated for C3oH₂5BrFN₇0₂.l.lHC1.1.6H₂0: C, 52.72; H, 4-32; N, 14.35; Cl, 5.71; Found: C, 52.79; H,

4.25; N, 14.28; Cl, 5.52.

Scheme 338 o

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-cyclopentyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (338e)

Step-1: Préparation of ethyl 2-(4-amino-6-(cyclopent-l-en-l-yl)-9H-pyrimido[4,5-b]indol-9yl)acetate (338a) and 2-(4-amino-6-(cyclopent-l-en-l-yl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (338b)

580

Compound 338a was prepared and purifïed according to the procedure reported in step-1 of scheme-62, from ethyl 2-(4-amino-6-bromo-9H-pyrimido[4,5-b]indol-9-yl)acetate (48a) (200 mg, 0.573 mmol) in dioxane (18 mL) using cyclopent-l-en-l-ylboronic acid (96 mg, 0.859 mmol), césium carbonate (280 mg, 0.859 mmol) in water (2.2 mL), Pd(PPh3)2Ch (80 mg, 0.115 mmol) to give a mixture of ethyl 2-(4-amino-6-(cyclopent-l-en-l-yl)-9H-pyrimido[4,5b]indol-9-yl)acetate (338a) and 2-(4-amino-6-(cyclopent-l-en-l-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (338b) which was used as such for the next step; MS (ES+): 309.10 & 337.20 (M+l).

Step-2: ethyl 2-(4-amino-6-cyclopentyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (338c) and 2(4-amino-6-cyclopentyl-9H-pÿrimido[4,5-b]indol-9-yl)acetic acid (338d)

To a solution of mixture of ethyl 2-(4-amino-6-(cyclopent-l-en-l-yl)-9H-pyrimido[4,5b]indol-9-yl)acetate (338a) and 2-(4-amino-6-(cyclopent-l-en-l-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (338b) from step-1 above (0.097 g, 0.287 mmol) in ethyl acetate (5 mL), éthanol (5 mL) and MeOH (5 mL) was treated with palladium (0.046 g, 0.043 mmol) and hydrogenated for 29 h. Reaction mixture was filtered, washed with ethyl acetate/methanol (9:1), and concentrated to obtained residue containing mixture of Ethyl 2(4-amino-6-cyclopentyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (338c) and 2-(4-amino-6cyclopentyl-9H-pyrimido[4,5-b]indol-9-yI)acetic acid (338d) (206 mg) which was used as such for next step; MS (ES+): 311.10 & 339.20 (M+l).

Step-3: Préparation of 2-(4-amino-6-cyclopentyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (338d)

Compound 338d was prepared according to the procedure reported in step-4 of scheme-17, from mixture of Ethyl 2-(4-amino-6-cyclopentyl-9H-pyrimido[4,5-b]indol-9-yI)acetate (338c) and 2-(4-amino-6-cyclopentyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (338d) (97 mg, 0.287 mmol) in THF (3 mL) and methanol (3 mL) using a solution of lithium hydroxide hydrate (73.7 mg, 1.722 mmol) in water (3 mL) and stirring at RT for 32 h. This gave after workup 2-(4-amino-6-cyclopentyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (338d) as a gray solid (56 mg) which was used as such for the next step; MS (ES+): 311.20 (M+l).

Step-4: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-cyclopentyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (338e)

Compound 338e was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-cyclopentyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (338d) (52 mg, 0.168 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5

581 methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (66.9 mg, 0.201 mmol), HATU (127 mg, 0.335 mmol), DIPEA (0.117 mL, 0.670 mmol) and stirring at RT for 19 h. This gave after workup as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6-cyclopentyl-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (338e) (22 mg, 22% yield) HCl sait as a light brown solid; 'H NMR (300 MHz, DMSO-îZ₆) δ 10.76 (s, 1H, D₂O exchangeable), 8.61 (s, 3H, D₂O exchangeable), 8.59 (s, 1H), 8.37 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.59 (d, J= 8.5 Hz, 1H), 7.48 - 7.43 (m, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.69 (d, J= 17.3 Hz, 1H), 5.35 (d, J= 17.3 Hz, 1H), 4.35 (dd, J= 9.0, 6.0 Hz, 1H), 3.69 (dd, J= 5.6, 2.4 Hz, 1H), 3.20 - 3.03 (m, 1H), 2.59 - 2.38 (m, 1H), 2.19-1.55 (m, 9H), 1.30 (s, 3H), 1.05 - 0.97 (m, 1H), 0.95 - 0.88 (m, 1H); MS (ES+): 588.20 & 590.20 (M+l); MS (ES-): 586.10 & 588.10 (M-l).

Scheme 339

339e 339f 339g

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(methoxymethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (339g) Step-1: Préparation of 2,2,2-trifluoro-N-(2-iodo-4-(methoxymethyl)phenyl)acetamide (339b) Compound 339b was prepared according to the procedure reported in step-1 of scheme-46, from 2-iodo-4-(methoxymethyl)aniline (339a)(6.538 g, 24.85 mmol; CAS # 1697458-72-2) in DCM (40 mL) using triethylamine (8.66 mL, 62.1 mmol), trifluoroacetic acid anhydride (5.18 mL, 37.3 mmol) and stirring at RT for 20 h. This gave after workup 2,2,2-trifluoro-N(2-iodo-4-(methoxymethyl)phenyl)acetamide (339b) (10.39 g) as a yellow solid and was used as such for next step; MS (ES-): 358.00 (M-l).

Step-2: Préparation of 2-amino-5-(methoxymethyl)-lH-indole-3-carbonitrile (339c)

582

Compound 339c was prepared according to the procedure reported in step-1 of scheme-11, from 2,2,2-trifluoro-N-(2-iodo-4-(methoxymethyl)phenyl)acetamide (339b) (8.92 g, 24.85 mmol) in DMSO (30 mL) using malononitrile (1.970 g, 29.8 mmol), L-proline (0.572 g, 4.97 mmol), Cul (0.473 g, 2.485 mmol), and K2CO3 (6.87 g, 49.7 mmol) in water (30 mL) to afford after workup and purification as described in step-1 of scheme-328, 2-amino-5(methoxymethyl)-lH-indole-3-carbonitrile (339c) (2.68 g, 54% yield) as a yellow solid; *H NMR (300 MHz, DMSO-t/ô) δ 10.69 (s, 1H), 7.11 - 7.04 (m, 2H), 6.88 - 6.82 (m, 1H), 6.75 (s, 2H), 4.38 (s, 2H), 3.24 (s, 3H); MS (ES+): 202.10 (M+l).

Step-3: Préparation of 6-(methoxymethyl)-9H-pyrimido[4,5-b]indol-4-amine (339d) Compound 339d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-5-(methoxymethyl)-lH-indole-3-carbonitrile (339c)(2.55 g, 12.67 mmol) in éthanol (60 mL) using formamidine acetate (10.66 g, 101 mmol) and refluxing for 19 h. This gave after workup 6-(methoxymethyl)-9H-pyrimido[4,5-b]indoI-4-amine (339d) (910 mg) as a light brown solid which was used as such for the next step; *H NMR (300 MHz, DMSO-dô) δ 11.81 (s, 1H), 8.27 (s, 1H), 8.23 (s, 1H), 7.40 (d, J= 8.2 Hz, 1H), 7.32 (dd, J= 8.2, 1.5 Hz, 1H), 7.14 (s, 2H), 4.51 (s, 2H), 3.29 (s, 3H); MS (ES+): 229.10 (M+l).

Step-4: Préparation of ethyl 2-(4-amino-6-(methoxymethyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (339e)

Compound 339e was prepared according to the procedure reported in step-1 of scheme-1, from 6-(methoxymethyl)-9H-pyrimido[4,5-b]indol-4-amine (339d) (850 mg, 3.72 mmol) in DMF (20 mL) using ethyl 2-bromoacetate (0.454 mL, 4.10 mmol), CS2CO3 (3033 mg, 9.31 mmol) and stirring at RT for 26 h. This gave after workup ethyl 2-(4-amino-6(methoxymethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (339e) (780 mg) as a light-yellow solid; Ή NMR (300 MHz, DMSO-ri₆) δ 8.35 - 8.32 (m, 1H), 8.28 (s, 1H), 7.56 (d, J= 8.3 Hz, 1H), 7.38 (dd, J= 8.4, 1.5 Hz, 1H), 7.30 (s, 2H), 5.23 (s, 2H), 4.53 (s, 2H), 4.14 (q, J= 7.1 Hz, 2H), 3.30 (s, 3H), 1.20 (t, J= 7.1 Hz, 3H); MS (ES+): 315.15 (M+l).

Step-5: Préparation of 2-(4-amino-6-(methoxymethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (339f)

Compound 339f was prepared according to the procedure reported in step-4 of scheme-17, from ethyl 2-(4-amino-6-(methoxymethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (339e) (600 mg, 1.909 mmol) in THF (8 mL) and methanol (8 mL) using a solution of lithium hydroxide hydrate (490 mg, 11.45 mmol) in water (8 mL) and stirring at RT for 19 h. This gave after workup 2-(4-amino-6-(methoxymethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (339f) (1.205 g) as a white solid which was used as such for next step; MS (ES+): 287.10 (M+l).

583

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(methoxymethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (339g)

Compound 339g was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(methoxymethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (339f) (0.23 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (4a) (0.088 g, 0.276 mmol), HATU (0.175 g, 0.460 mmol), DIPEA (0.160 mL, 0.92 mmol) and stirring at RT ovemight. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6-(methoxymethyI)9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (339g) (83 mg, 66% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-tZe) δ 10.76 (s, 1H, D₂O exchangeable), 8.75 (s, 2H, D₂O exchangeable), 8.64 (s, 1H), 8.52 (s, 1H), 8.00 (d, J= 8.2 Hz, 1H), 7.80 - 7.63 (m, 2H), 7.54 (d, J= 8.4 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.77 (d, J= 17.4 Hz, 1H), 5.42 (d, J= 17.3 Hz, 1H), 4.56 (s, 2H), 4.47 - 4.35 (m, 1H), 3.98-3.83 (m, 1H), 3.32 (s, 3H), 2.40-2.10 (m, 2H), 1.99-1.82 (m, 1H), 1.190.96 (m, 1H), 0.89 - 0.65 (m, 1H); MS (ES+): 550.10 & 552.15 (M+l); MS (ES-): 548.10 & 550.10 (M-l); Analysis calculated for C₂5H₂₄BrN₇O3.1.15HC1.1.0H₂O: C, 49.20; H, 4.48; N, 16.06; Cl, 6.68; Found: C, 49.30; H, 4.44; N, 15.99; Cl, 6.56.

Scheme 340

^339f 340a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(methoxymethyl)-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (340a)

Compound 340a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(methoxymethyI)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (339f) (0.23 mmol) in DMF (10 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (8a) (92 mg, 0.276 mmol), HATU (175 mg, 0.460

584 mmol), DIPEA (0.160 mL, 0.920 mmol) and stirring at RT ovemight. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6-(methoxymethyl)9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-bromopyridin-2-yl)-5-methyI-2azabicyclo[3.1.0]hexane-3-carboxamide (340a) (80 mg, 62% yield) HCl sait as a white solid;

Ή NMR (300 MHz, DMSOX) δ 10.77 (s, 1H, D₂Oexchangeable), 8.69 (s, 2H, D₂Oexchangeable), 8.62 (s, 1H), 8.50 (s, 1H), 8.00 (d, J= 8.2 Hz, 1H), 7.75 - 7.64 (m, 2H), 7.53 (dd, J= 8.5, 1.5 Hz, 1H), 7.31 (d, 7=7.7 Hz, 1H), 5.72 (d, J= 17.4 Hz, 1H), 5.37 (d, J= 17.3 Hz, 1H), 4.56 (s, 2H), 4.37 (dd, J= 9.1, 6.0 Hz, 1H), 3.69 (dd, J= 5.5, 2.4 Hz, 1H), 3.31 (s, 3H), 2.55-2.41 (m, 1H), 1.98 (dd, J= 13.2, 5.9 Hz, 1H), 1.30 (s, 3H), 1.01 (t, J= 5.5 Hz, 10 1H), 0.93 (dd, J= 5.4, 2.4 Hz, 1H); MS (ES+): 564.10 & 566.10 (M+l); MS (ES-): 562.10 &

564.10 (M-l); Analysis calculated for C₂₆H₂6BrN₇O₃.1.0HC1.1.5H₂O: C, 49.73; H, 4.82; N, 15.61; Cl, 5.65; Found: C, 49.79; H, 4.86; N, 15.71; Cl, 5.38.

Scheme 341

341b 341c

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(3-methoxyphenyl)-8-methyl-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (341c)

Step-1: Préparation of terZ-butyl 2-(4-amino-6-(3-methoxyphenyl)-8-methyl-9Hpyrimido[4,5-b]indol-9-yl)acetate (341a)

Compound 341a was prepared and purified according to the procedure reported in step-1 of scheme-62, from ZerZ-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-921436

585 yl)acetate (192e) (200 mg, 0.511 mmol) in dioxane (18 mL) using (3-methoxyphenyl)boronic acid (117 mg, 0.767 mmol), césium carbonate (250 mg, 0.767 mmol) in water (2.2 mL), Pd(PPh3)2C12 (71.8 mg, 0.102 mmol) to afford tert-butyl 2-(4-amino-6-(3-methoxyphenyl)-8methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (341a) (110 mg, 51% yield) as a white solid; MS (ES+): 419.20 (M+l).

Step-2: Préparation of 2-(4-amino-6-(3-methoxyphenyl)-8-methyl-9H-pyrimido[4,5-b]indol9-yl)acetic acid (341b)

Compound 341b was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-6-(3-methoxyphenyl)-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetate (341a) (105 mg, 0.251 mmol) in DCM (10 mL) using TFA (0.483 mL, 6.27 mmol) and stirring at RT for 27 h. This gave after workup 2-(4-amino-6-(3-methoxyphenyl)-8methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (341b) and was used as such for the next step; MS (ES+): 363.10 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(3-methoxyphenyl)-8-methyl-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (341c)

Compound 341c was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(3-methoxyphenyl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (341b) (0.125 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (0.048 g, 0.150 mmol), HATU (0.095 g, 0.250 mmol), DIPEA (0.109 mL, 0.625 mmol) and stirring at RT for 22 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6-(3methoxyphenyl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (341c) (26 mg, 33% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.83 (s, 1H, D₂O exchangeable), 8.69 (s, 2H, D₂O exchangeable), 8.62 (s, 1H), 8.59 - 8.56 (m, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.64 (s, 1H), 7.45 - 7.35 (m, 3H), 7.31 (d, J= 7.7 Hz, 1H), 7.01 - 6.90 (m, 1H), 5.94 (d, J= 18.0 Hz, 1H), 5.66 (d, J= 17.9 Hz, 1H), 4.43 (dd, J= 9.0, 5.7 Hz, 1H), 3.98 - 3.89 (m, 1H), 3.85 (s, 3H), 2.77 (s, 3H), 2.45-2.29 (m, 1H), 2.28-2.14 (m, 1H), 2.01 - 1.86 (m, 1H), 1.15 - 1.03 (m, 1H), 0.78 - 0.56 (m, 1H); MS (ES+): 626.10 & 628.20 (M+l); MS (ES-): 624.10 & 626.10 (M-l); Analysis calculated for C₃iH₂₈BrN₇03.1.0HC1.2.0H₂0: C, 53.27; H, 4.76; N, 14.03; Found: C, 53.14; H, 4.60; N, 13.87.

Scheme 342

586

341b

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(3-methoxyphenyl)-8-methyl-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicycIo[3.1.0]hexane-3carboxamide (342a)

Compound 342a was prepared according to the procedure reported in step-3 of scheme-1, from 2-(4-amino-6-(3-methoxyphenyl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (341b) (0.125 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (49.9 mg, 0.150 mmol), HATU (95 mg, 0.250 mmol), DIPEA (0.109 mL, 0.625 mmol) and stirring at RT for 22 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6-(3methoxyphenyl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (342a) (27 mg, 34% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î7₆) δ 10.82 (s, 1H, D₂O exchangeable), 8.57 (s, 1H), 8.56 - 8.48 (m, 4H, 3H D₂O exchangeable ), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.61 (s, 1H), 7.43 - 7.35 (m, 3H), 7.31 (d, J= 7.7 Hz, 1H), 7.01 - 6.89 (m, 1H), 5.88 (d, J= VIS) Hz, 1H), 5.61 (d, J= 17.8 Hz, 1H), 4.39 (dd, J= 8.9, 6.2 Hz, 1H), 3.85 (s, 3H), 3.74 -3.68 (m, 1H), 2.74 (s, 3H), 2.58 - 2.42 (m, 1H), 1.98 (dd, J= 13.2, 6.1 Hz, 1H), 1.31 (s, 3H), 1.02 (t, J= 5.4 Hz, 1H), 0.87 - 0.82 (m, 1H); MS (ES+): 640.20 & 642.15 (M+l); MS (ES-): 638.10 & 640.10 (M-l); Analysis calculated for C32H3oBrN703.1.0HC1.1.75H₂0: C, 54.25; H, 4.91; N, 13.84; Found: C, 54.21; H, 4.81; N, 13.72.

Scheme 343

587

Me

308b 343a

Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2carboxamide (343a)

Compound 343a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5-b]indol9-yl)acetic acid (308b) (75 mg, 0.162 mmol) in DMF (5 mL) using TFA sait of (2S,4R)-N(6-chloropyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (320b) (60.3 mg, 0.162 mmol), HATU (74.0 mg, 0.195 mmol), DIPEA (105 mg, 0.811 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (2S,4R)-l-(2-(4amino-8-methyl-6-(2-methylpyrimidin-5-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6chloropyridin-2-yl)-4-fluoro-4-methylpyrrolidine-2-carboxamide (343a) (43 mg, 45% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-Jd) δ (a mixture of two rotamers) 11.37 and 11.01 (2s, 1H, D₂O exchangeable), 9.24 and 9.23 (2s, 2H), 9.04 (bs, 2H, D₂O exchangeable), 8.80 and 8.77 (2s, 1H), 8.68 (s, 1H), 7.97 (d, J = 8.2 Hz, 1H), 7.87 - 7.75 (m, 2H), 7.18 (d, J = 7.7 Hz, 1H), 5.77 (d, J = 18.0 Hz, 1H), 5.56 (d, J = 17.7 Hz, 1H), 4.70 (dd, J = 9.7, 7.5 Hz, 1H), 4.37 - 4.22 (m, 1H), 3.98 - 3.93 (m, 1H), 2.77 (s, 3H), 2.70 (s, 3H), 2.65 2.56 (m, 1H), 2.22 - 1.97 (m, 1H), 1.64 (d, J = 21.1 Hz, 3H); ¹⁹F NMR (282 MHz, DMSO-ifc) δ -139.90; MS (ES+): 588/590 (M+l); (ES-): 586/588 (M-l); Analysis calculated for C₂₉H₂₇C1FN₉O₂.1.35HC1.3.25H₂O: C, 50.06; H, 5.05; Cl, 11.97; N, 18.12; Found: C, 50.03; H, 4.93; Cl, 12.07; N, 17.91.

Scheme 344

588

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indoI-9-yI)acetyl)-N-(6-bromo-4-methoxypyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (344c)

Step-1 : Préparation of (lR,3S,5R)-/erZ-butyl 3-((6-bromo-4-methoxypyridin-2yI)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (344a)

An aliquot of l-chloro-N,N,2-trimethylprop-l-en-l-amine (0.090 mL, 0.677 mmol) was added drop wise to a solution of (lR,3S,5R)-2-(ZerZ-butoxycarbonyl)-2azabicyclo[3.1.0]hexane-3-carboxylic acid (259a) (140 mg, 0.616 mmol) in DCM (10 mL) at 0 °C and the mixture was stirred at 0 °C for 1 h, followed by addition of 6-bromo-4methoxypyridin-2-amine (125 mg, 0.616 mmol), DIPEA (159 mg, 1.231 mmol) and stirring at RT for 16 h. The reaction mixture was diluted with DCM (15 mL), washed with H₂O (20 mL><2), brine (20 mL) dried, filtered, concentrated and the residue obtained was purified using flash column chromatography [SiO₂ gel, eluting with EtOAc in hexane from 0-20%] to provide ( 1 R,3S,5R)-terZ-butyl 3-((6-bromo-4-methoxypyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (344a) (155 mg, 61% yield) as a pale-yellow solid; ’H NMR (300 MHz, DMSO-J₆) δ 10.74 (s, 1H), 7.69 (s, 1H), 6.99 (d, J= 2.0 Hz, 1H), 4.26 4.01 (m, 1H), 3.85 (s, 3H), 3.47-3.35 (m, 1H), 2.37-2.18 (m, 1H), 2.18-2.01 (m, 1H), 1.68-1.49 (m, 1H), 1.33 (d, J= 46.4 Hz, 9H), 0.83-0.65 (m, 1H), 0.47 - 0.31 (m, 1H); MS (ES+): 412/414 (M+l); (ES-): 410/412 (M-l).

589

Step-2: Préparation of (lR,3S,5R)-N-(6-bromo-4-methoxypyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (344b)

A mixture of (lR,3S,5R)-to7-butyl 3-((6-bromo-4-methoxypyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (344a) (155 mg, 0.376 mmol), 4 M HCl in dioxane (0.94 mL, 3.76 mmol) were suspended in DCM (5 mL), stirred at RT for 16 h and concentrated in vacuum to dryness. The residue was triturated with ether and the solid obtained was collected by filtration to provide the HCl sait of (lR,3S,5R)-N-(6-bromo-4methoxypyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (344b) (130 mg, 99 % yield) as a white solid; *H NMR (300 MHz, DMSO-dô) δ 11.32 (s, 1H), 10.51 (bs, J= 6.9 Hz, 10 1H), 9.18 (bs, 1H), 7.60 (d, J= 2.0 Hz, 1H), 7.09 (d, J= 2.0 Hz, 1H), 4.29 - 4.04 (m, 1H),

3.87 (s, 3H), 3.39-3.25 (m, 1H), 2.69-2.54 (m, 1H), 2.16-1.96 (m, 1H), 1.90-1.73 (m, 1H), 0.95 - 0.65 (m, 2H); MS (ES+): 312/314 (M+l); (ES-): 310/312 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-4-methoxypyridin-2-yl)-215 azabicyclo[3.1.0]hexane-3-carboxamide (344c)

Compound 344c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (lR,3S,5R)-N-(6-bromo-4-methoxypyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (344b) (59.7 mg, 0.171 mmol) in DMF (5 mL) using HCl sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (75 mg, 0.171 mmol), HATU (98 mg, 0.257 mmol), DIPEA (111 mg, 0.856 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyI)-N-(6-bromo-4-methoxypyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3• carboxamide (344c) (27 mg, 34% yield) HCl sait as a white solid; ’H NMR (300 MHz,

DMSO-dô) δ 10.77 (s, 1H, D₂O exchangeable), 9.03 (bs, 2H, D₂O exchangeable), 8.83 (s,

1H), 8.72 (s, 1H), 7.64 (d, J = 1.7 Hz, 1H), 7.60 (d, J = 2.0 Hz, 1H), 6.95 (d, J = 2.0 Hz, 1H), 5.98 (d, J = 18.1 Hz, 1H), 5.69 (d, J = 18.0 Hz, 1H), 4.49 - 4.37 (m, 1H), 3.96 - 3.90 (m, 1H), 3.82 (s, 3H), 2.78 (s, 3H), 2.41 - 2.27 (m, 1H), 2.26 - 2.13 (m, 1H), 2.02 - 1.83 (m, 1H), 1.15 - 1.01 (m, 1H), 0.78 - 0.59 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-é7₆) δ -58.86; MS (ES+):

618/620 (M+l); (ES-): 616/618 (M-l); Analysis calculated for

C₂6H₂₃BrF3N₇O3.1.1HC1.1.75H₂O: C, 45.26; H, 4.03; Cl, 5.65; N, 14.21; Found: C, 45.34; H, 3.95; Cl, 5.74; N, 14.16.

Scheme 345

590

DIPEA

CF₃ ^345b 345c

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-(difluoromethoxy)pyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (345c)

Step-1 : Préparation of (lR,3S,5R)-tert-butyl 3-((6-(difluoromethoxy)pyridin-2yl)carbamoyl)-2-azabicyclo[3.1,0]hexane-2-carboxylate (345a)

Compound 345a was prepared and purified according to the procedure reported in step-1 of scheme-344, from (lR,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3carboxylic acid (259a) (177 mg, 0.781 mmol) in DCM (5 mL) using l-chloro-N,N,210 trimethylprop-1 -en-1 -amine (0.114 mL, 0.859 mmol), 6-(difluoromethoxy)pyridin-2-amine (125 mg, 0.781 mmol) and DIPEA (202 mg, 1.561 mmol) to afford (lR,3S,5R)-ter/-butyl 3(6-(difluoromethoxy)pyridin-2-ylcarbamoyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (345a) (170 mg, 59% yield) as a pale-yellow solid; *H NMR (300 MHz, DMSO-dô) δ 10.44 (s, 1H), 8.04 - 7.85 (m, 2H), 7.57 (s, 1H), 6.88 - 6.64 (m, 1H), 4.34 - 4.05 (m, 1H), 3.35 (s,

1H), 2.38 - 2.21 (m, 1H), 2.21 - 2.02 (m, 1H), 1.71 - 1.53 (m, 1H), 1.32 (s, 9H), 0.84 - 0.64 (m, 1H), 0.50 - 0.33 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-^) δ -86.36; MS (ES+): 370 (M+l); (ES-): 368 (M-l).

Step-2: Préparation of (lR,3S,5R)-N-(6-(difluoromethoxy)pyridin-2-yl)-2azabicyclo[3.1,0]hexane-3-carboxamide (345b)

Compound 345b was prepared according to the procedure reported in step-2 of scheme-344, from (lR,3S,5R)-ter/-butyl 3-((6-(difluoromethoxy)pyridin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (345a) (165 mg, 0.447 mmol) in DCM (5 mL) using 4 M HCl in dioxane (1.117 mL, 4.47 mmol) and stirring at RT for 16 h. This gave after work

591 up HCl sait of (lR,3S,5R)-N-(6-(difluoromethoxy)pyridin-2-yl)-2-azabicyclo[3.1.0]hexane3-carboxamide (345b) (123 mg, 90% yield) as a white solid^IH NMR (300 MHz, DMSO-ùfc) δ 11.08 (s, 1H), 7.98 (t, J = 8.0 Hz, 1H), 7.86 (d, J= 8.1 Hz, 1H), 7.58 (t, J= 72.8 Hz, 1H), 6.88 (d, J =7.9 Hz, 1H), 4.34-4.08 (m, 1H), 3.33 - 3.18 (m, 1H), 2.66-2.54 (m, 1H),2.19 - 1.98 (m, 1H), 1.91 - 1.62 (m, 1H), 0.91 - 0.76 (m, 2H); ¹⁹F NMR (282 MHz, DMSO-J₆) δ -86.32; MS (ES+): 270 (M+l); (ES-): 268 (M-l)

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-(difluoromethoxy)pyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (345c)

Compound 345c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (lR,3S,5R)-N-(6-(difluoromethoxy)pyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (345b) (52.3 mg, 0.171 mmol) in DMF (5 mL) using TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (75 mg, 0.171 mmol), HATU (98 mg, 0.257 mmol), DIPEA (111 mg, 0.856 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (1R,3 S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido [4,5b]indol-9-yl)acetyl)-N-(6-(difluoromethoxy)pyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (345c) (56 mg, 57% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-î/ô) δ 10.54 (s, 1H, D₂O exchangeable), 8.86 (bs, 2H, D₂O exchangeable), 8.82 (s, 1H), 8.68 (s, 1H), 7.91 - 7.80 (m, 2H), 7.63 (d, J = 1.7 Hz, 1H), 7.50 (t, J = 73.0 Hz, 1H), 6.82 - 6.67 (m, 1H), 5.99 (d, J = 18.0 Hz, 1H), 5.70 (d, J = 18.0 Hz, 1H), 4.49 (dd, J = 9.0, 5.7 Hz, 1H), 3.99 - 3.91 (m, 1H), 2.79 (s, 3H), 2.42 - 2.30 (m, 1H), 2.28 - 2.14 (m, 1H), 2.03 1.87 (m, 1H), 1.17 - 1.04 (m, 1H), 0.70 (m, J = 5.2, 2.4 Hz, 1H); ¹⁹F NMR (282 MHz, DMSO-îZô) δ -58.82, -86.30, -86.40; MS (ES+): 576 (M+l); (ES-): 574 (M-l); Analysis calculated for C₂₆H₂₂F₅N₇O3.HC1.1.5H₂O: C, 48.87; H, 4.10; Cl, 5.55; N, 15.34; Found: C, 48.78; H, 4.12; Cl, 5.53; N, 15.26.

Scheme 346

592

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1,0]hexane-3-carboxamide (346g)

Step-1 : Préparation of N-(4-bromo-2-methyl-6-(trifluoromethyl)pyridin-3-yl)-2,2,2trifluoroacetamide (346b)

Compound 346b was prepared according to the procedure reported in step-1 of scheme-46, from 4-bromo-2-methyI-6-(trifluoromethyl)pyridin-3-amine (346a)(6.16 g, 24.15 mmol; CAS # 1990462-02-6) in DCM (25 mL) using triethylamine (4.15 g, 41.1 mmol), trifluoroacetic acid anhydride (7.61 g, 36.2 mmol) to afford after workup N-(4-bromo-2-methyl-6(trifluoromethyl)pyridin-3-yl)-2,2,2-trifluoroacetamide (346b) (8.08 g, 95% yield) as a paleyellow solid and was used as such for next step; *H NMR (300 MHz, DMSO-îZô) δ 11.84 (s, 1H), 8.32 (s, 1H), 2.51 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -66.43, -74.09; MS (ES+): 351/353 (M+l); (ES-): 349/351 (M-l).

Step-2: Préparation of 2-amino-7-methyl-5-(trifluoromethyl)-lH-pyrrolo[2,3-c]pyridine-3carbonitrile (346c)

Compound 346c was prepared and purified according to the procedure reported in step-1 of scheme-11, from N-(4-bromo-2-methyl-6-(trifluoromethyl)pyridin-3-yl)-2,2,2trifluoroacetamide (346b) (8.08 g, 23.02 mmol) in DMSO (30 mL) using malononitrile (1.825 g, 27.6 mmol), L-proline (0.530 g, 4.60 mmol), Cul (0.438 g, 2.302 mmol), a solution of K2CO3 (6.36 g, 46.0 mmol) in water (30 mL) to afford 2-amino-7-methyl-5(trifluoromethyl)-lH-pyrrolo[2,3-c]pyridine-3-carbonitrile (346c) (3.76 g, 68% yield) as a yellow solid; Ή NMR (300 MHz, DMSO-î7₆) δ 11.68 (s, 1H), 7.40 (s, 1H), 7.27 (s, 2H), 2.57 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-tfc) δ -64.65; MS (ES+): 241 (M+l); (ES-): 239 (M-l).

593

Step-3: Préparation of 8-methyl-6-(tnfluoromethyl)-9H-pyndo[4',3':4,5]pyrrolo[2,3d]pyrimidin-4-amine (346d)

Compound 346d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-7-methyl-5-(trifluoromethyl)-lH-pynOlo[2,3-c]pyridine-3-carbonitrile (346c) (3.76 g, 15.65 mmol) in éthanol (10 mL) using trimethoxymethane (16.61 g, 157 mmol), NKUOAc (3.62 g, 47.0 mmol) and heating at 90 °C for 16 h. This gave after workup 8methyl-6-(trifluoromethyl)-9H-pyrido[4^,,3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (346d) (3.53 g, 84% yield) as apale-yellow solid; *HNMR (300 MHz, DMSO-îZô) δ 12.66 (s, 1H), 8.78 (s, 1H), 8.40 (s, 1H), 7.72 (bs, 2H), 2.79 (s, 3H); ¹⁹F NMR (282 MHz, DMSO) -d₆ δ -63.78; MS (ES+): 268 (M+l); (ES-): 266 (M-l).

Step-4: Préparation of ZerZ-butyl 2-(4-amino-8-methyl-6-(trifIuoromethyl)-9Hpyrido^'jS'^jSlpyrroloPjS-djpyrimidin^-yljacetate (346e)

Compound 346e was prepared according to the procedure reported in step-1 of scheme-1, from 8-methyl-6-(trifluoromethyl)-9H-pyrido[4',3':4,5]pyrrplo[2,3-d]pyrimidin-4-amine (346d) (3.53 g, 13.21 mmol) in DMF (20 mL) using ZerZ-butyl 2-bromoacetate (3.09 g, 15.85 mmol), CS2CO3 (8.61 g, 26.4 mmol) and stirring at RT for 16 h. This gave after workup ZerZbutyl 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin9-yl)acetate (346e) (3.46 g, 69% yield) as a pale-yellow solid; *H NMR (300 MHz, DMSOd₆) δ 8.86 (s, 1H), 8.45 (s, 1H), 7.91 (bs, 2H), 5.44 (s, 2H), 2.88 (s, 3H), 1.44 (s, 9H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -64.14; MS (ES+): 382 (M+l); (ES-): 380 (M-l).

Step-5 : Préparation of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (346f)

Compound 346f was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetate (346e) (366 mg, 0.960 mmol) using TFA (1094 mg, 9.60 mmol) in DCM (5 mL) to afford after workup TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (346f) (0.280 g, 66% yield) as a pale-yellow solid; ’H NMR (300 MHz, DMSO-î/₆) δ 8.88 (s, 1H), 8.48 (s, 1H), 8.04 (bs, 2H), 5.47 (s, 2H), 2.91 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -64.16, -74.52; MS (ES+): 326 (M+l), (ES-): 324 (M-l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo [3.1.0]hexane-3 -carboxamide (346g)

594

Compound 346g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[4',3':4,5]pyrroIo[2,3d]pyrimidin-9-yl)acetic acid (346f) (60 mg, 0.137 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (43.5 mg, 0.137 mmol), HATU (62.3 mg, 0.164 mmol), DIPEA (88 mg, 0.683 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (346g) (72 mg, 89% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-d₆) δ 10.85 (s, 1H, D₂O exchangeable), 8.99 (s, 1H), 8.95 (bs, 2H, D₂O exchangeable), 8.71 (s, 1H), 8.01 (d, J = 8.1 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 6.03 (d, J = 18.0 Hz, 1H), 5.73 (d, J = 17.9 Hz, 1H), 4.46 - 4.40 (m, 1H), 4.00 - 3.92 (m, 1H), 2.43 - 2.30 (m, 1H), 2.28 - 2.12 (m, 1H), 2.04 - 1.82 (m, 1H), 1.10 (m, J = 8.8, 5.4 Hz, 1H), 0.80 - 0.59 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-d₆) δ -64.39; MS (ES+): 589/591 (M+l); (ES-): 587/589 (M-l);

Analysis calculated for C₂₄H₂oBrF₃N₈0₂.l.lHC1.2H₂0: C, 43.31; H, 3.80; Cl, 5.86; N, 16.84; Found: C, 43.24; H, 3.73; Cl, 5.97; N, 16.73.

Scheme 347

CF₃

346f ^347a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H20 pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (347a)

Compound 347a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetic acid (346f) (60 mg, 0.137 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (45.4 mg, 0.137 mmol), HATU (62.3 mg, 0.164 mmol), DIPEA (88 mg, 0.683 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme

595

299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (347a) (63 mg, 76 % yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/ô) δ 10.84 (s, 1H, D2O exchangeable), 8.95 (s, 1H), 8.66 (bs, 2H, D2O exchangeable), 8.63 (s, 1H), 8.02 (d, J = 8.2 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.95 (d, J = 18.0 Hz, 1H), 5.68 (d, J = 17.9 Hz, 1H), 4.39 (dd, J = 9.1, 6.2 Hz, 1H), 3.77 - 3.73 (m, 1H), 2.93 (s, 3H), 2.50 - 2.41 (m, 1H), 1.99 (dd, J= 13.3, 6.1 Hz, 1H), 1.32 (s, 3H), 1.07 - 0.98 (m, 1H), 0.96 - 0.82 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-cfc) δ -64.32; MS (ES+): 603/605 (M+l); (ES-): 601/603 (M-l); Analysis calculated for C25H22BrF3N₈O2.0.95HC1.1.75H2O: C, 44.85; H, 3.98; Cl, 5.03; N, 16.74; Found: C, 44.76; H, 3.80; Cl, 5.06; N, 16.60.

Scheme 348

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-(trifluoromethyl)pyrazin-2-yl)-2-azabicyclo[3.1,0]hexane-3carboxamide (348c)

Step-1: Préparation of (lR,3S,5R)-tertebutyl 3-((6-(trifluoromethyl)pyrazin-2-yl)carbamoyl)2-azabicyclo[3.1,0]hexane-2-carboxylate (348a)

Compound 348a was prepared and purified according to the procedure reported in step-1 of scheme-344, from (lR,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3carboxylic acid (259a) (174 mg, 0.766 mmol) in DCM (10 mL) using l-chloro-N,N,2trimethylprop-1-en-1-amine (0.112 mL, 0.843 mmol), 6-(trifluoromethyl)pyrazin-2-amine (125 mg, 0.766 mmol), DIPEA (198 mg, 1.533 mmol) to afford (lR,3S,5R)-tert-butyl 3-((6

596 (trifluoromethyl)pyrazin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (348a) (63 mg, 22% yield) as apale-yellow solid; MS (ES+): 373 (M+l); (ES-): 371 (M-l).

Step-2: Préparation of (lR,3S,5R)-N-(6-(trifluoromethyl)pyrazin-2-yI)-2azabicyclo[3.1.0]hexane-3-carboxamide (348b)

Compound 348b was prepared according to the procedure reported in step-2 of scheme-344, from (lR,3S,5R)-fërt-butyl 3-((6-(trifluoromethyl)pyrazin-2-yl)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (348a) (63 mg, 0.169 mmol) in DCM (5 mL) using 4 M HCl in dioxane (0.423 mL, 1.692 mmol) to afford HCl sait of (lR,3S,5R)-N-(6(trifluoromethyl)pyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (348b) (42 mg, 80% yield) as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 11.88 (s, 1H), 9.58 (s, 1H), 8.98 (s, 1H), 4.25 (dd, J= 10.8, 7.8 Hz, 1H), 3.40 - 3.30 (m, 1H), 2.64 (dd, J= 12.8, 7.8 Hz, 1H), 2.23 - 2.04 (m, 1H), 1.90 - 1.74 (m, 1H), 0.93 - 0.76 (m, 2H); ¹⁹F NMR (282 MHz, DMSO-îZô) δ -66.42; MS (ES+): 273 (M+l); (ES-): 271 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifhioromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-(trifluoromethyl)pyrazin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (348c)

Compound 348c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (lR,3S,5R)-N-(6-(trifluoromethyl)pyrazin-2-yl)-2-azabicyclo[3.1.0]hexane3-carboxamide (348b) (42.3 mg, 0.137 mmol) in DMF (5 mL) using TFA sait of 2-(4-amino8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (88 mg, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6(trifluoromethyl)pyrazin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (348c) (58 mg, 73% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ 11.38 (s, 1H, D₂O exchangeable), 9.55 (s, 1H), 8.85 (s, 1H), 8.80 (s, 1H), 8.70 (bs, 2H, D₂O exchangeable), 8.64 (s, 1H), 7.61 (d, J = 1.7 Hz, 1H), 5.99 (d, J = 18.0 Hz, 1H), 5.71 (d, J = 17.9 Hz, 1H), 4.51 (dd, J = 9.0, 5.8 Hz, 1H), 4.01 - 3.91 (m, 1H), 2.79 (s, 3H), 2.46 - 2.34 (m, 1H), 2.34 - 2.22 (m, 1H), 2.03 - 1.89 (m, 1H), 1.16 - 1.03 (m, 1H), 0.81 - 0.66 (m, 1H); ¹⁹FNMR (282 MHz, DMSO-Jé) δ -58.78, -66.52; MS (ES+): 579 (M+l), (ES-): 577 (M-l); Analysis calculated for C₂₅H₂₀F₆N8O₂.l.lHCl.H₂O: C, 47.17; H, 3.66; Cl, 6.13; N, 17.60; Found: C, 47.00; H, 3.61; Cl, 6.09; N, 17.45.

Scheme 349

597

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-(trifluoromethyl)pyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (349c)

Step-1 : Préparation of (lR,3S,5R)-/eri-butyl 3-((6-(trifluoromethyl)pyridin-2-yl)carbamoyI)2-azabicyclo[3.1.0]hexane-2-carboxylate (349a)

Compound 349a was prepared according to the procedure reported in step-1 of scheme-345, from (lR,3S,5R)-2-(ter/-butoxycarbonyl)-2-azabicyclo[3.1,0]hexane-3-carboxylic acid (259a) (350 mg, 1.542 mmol) in DCM (5 mL) using l-chloro-N,N,2-trimethylprop-l-en-l10 amine (0.224 mL, 1.696 mmol), 6-(trifluoromethyl)pyridin-2-amine (250 mg, 1.542 mmol), DIPEA (399 mg, 3.08 mmol) to afford after work up and purification using flash column chromatography [SiCh gel, eluting with EtOAc in hexane from 0-15%] followed by purification using reverse phase column chromatography [Cl8 column (100 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%] (lR,3S,5R)-to7-butyl 3-((615 (trifluoromethyl)pyridin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (349a) (280 mg, 49% yield) as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 10.92 (s, 1H), 8.38 (d, J= 8.4 Hz, 1H), 8.09 (t, J= 8.1 Hz, 1H), 7.61 (d, J= 7.5 Hz, 1H), 4.36 - 4.03 (m, 1H), 3.53-3.37 (m, 1H), 2.39-2.23 (m, 1H), 2.23-2.02 (m, 1H), 1.74-1.52 (m, 1H), 1.37 (d, 9H), 0.89 - 0.65 (m, 1H), 0.53 - 0.31 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-d₆) δ -66.61; MS (ES+): 394 (M+Na); (ES-): 370 (M-l).

Step-2: Préparation of (lR,3S,5R)-N-(6-(trifluoromethyl)pyridin-2-yl)-2azabicycIo[3.1.0]hexane-3-carboxamide (349b)

598

Compound 349b was prepared according to the procedure reported in step-2 of scheme-344, from (lR,3S,5R)-terf-butyl 3-((6-(trifluoromethyl)pyridin-2-yI)carbamoyl)-2azabicyclo[3.1.0]hexane-2-carboxylate (349a) (280 mg, 0.754 mmol) in DCM (5 mL) using 4 M HCl in dioxane (1.885 mL, 7.54 mmol) to afford HCl sait of (lR,3S,5R)-N-(6(trifluoromethyI)pyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (349b) (230 mg, 99% yield) as a white solid; ‘HNMR (300 MHz, DMSO-J₆) δ 11.48 (s, 1H), 8.32 (d, J= 8.5 Hz, 1H), 8.17 (t, J= 8.0 Hz, 1H), 7.71 (d, J= 7.5 Hz, 1H), 4.38 - 4.07 (m, 1H), 3.41 - 3.29 (m, 1H), 2.62 (dd, 7= 12.8, 7.7 Hz, 1H), 2.27-2.05 (m, 1H), 1.98 - 1.72 (m, 1H), 0.98 0.58 (m, 2H); ¹⁹F NMR (282 MHz, DMSO-cfc) δ -66.61; MS (ES+): 272 (M+l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-(trifluoromethyl)pyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (349c)

Compound 349c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (lR,3S,5R)-N-(6-(trifluoromethyl)pyridin-2-yI)-2-azabicyclo[3.1.0]hexane3-carboxamide (349b) (42.1 mg, 0.137 mmol) in DMF (5 mL) using TFA sait of 2-(4-amino8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (1841) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (88 mg, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yI)acetyl)-N-(6(trifluoromethyl)pyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (349c) (54 mg, 68% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 10.96 (s, 1H, D₂O exchangeable), 8.80 (s, 1H), 8.77 (bs, 2H, D₂O exchangeable), 8.66 (s, 1H), 8.29 (d, J = 8.5 Hz, 1H), 8.05 (t, J = 8.0 Hz, 1H), 7.61 (s, 1H), 7.58 (d, J = 7.6 Hz, 1H), 5.99 (d, J = 18.0 Hz, 1H), 5.70 (d, J = 18.0 Hz, 1H), 4.49 (dd, J = 9.0, 5.7 Hz, 1H), 4.01 - 3.90 (m, 1H), 2.79 (s, 3H), 2.44 - 2.31 (m, 1H), 2.30 - 2.16 (m, 1H), 2.03 - 1.86 (m, 1H), 1.18 - 0.98 (m, 1H), 0.78 0.62 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-7₆) δ -58.80, -66.65; MS (ES+): 578 (M+l), (ES): 576 (M-l); Analysis calculated for C₂₆H₂iF₆N7O₂.1.1HC1.1.25H₂O: C, 48.79; H, 3.87; Cl, 6.09; N, 15.32; Found: C, 48.59; H, 3.85; Cl, 5.97; N, 15.33.

Scheme 350

599

346f ^CF3 _350a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrido [4',3 ' :4,5]pyrrolo [2,3 -d]pyrimidin-9-yI)acetyl)-N-(6-bromopyrazin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (350a)

Compound 350a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyI-6-(trifluoromethyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetic acid (346f) (60 mg, 0.137 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (261c) (45.6 mg, 0.137 mmol), HATU (62.3 mg, 0.164 mmol), DIPEA (88 mg, 0.683 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (350a) (57 mg, 69% yield) HCl sait as a white solid; ^!H NMR (300 MHz, DMSO-î/ô) δ 11.21 (s, 1H, D₂O exchangeable), 9.24 (s, 1H), 8.95 (s, 1H), 8.68 (bs, 2H, D₂O exchangeable), 8.64 (s, 1H), 8.53 (s, 1H), 5.96 (d, J = 18.0 Hz, 1H), 5.68 (d, J = 17.9 Hz, 1H), 4.41 (dd, J = 9.1, 6.2 Hz, 1H), 3.80 - 3.70 (m, 1H), 2.92 (s, 3H), 2.60 - 2.52 (m, 1H), 2.03 (dd, J= 13.2, 6.1 Hz, 1H), 1.32 (s, 3H), 1.09 - 0.99 (m, 1H), 0.96 - 0.86 (m, 1H). ¹⁹FNMR (282 MHz, DMSO-rid) δ -64.34; MS (ES+): 604/606 (M+l); (ES-): 602/604 (M-l); Analysis calculated for C24H21BrF3N9O2.L45HCl.2H2O: C, 41.58; H, 3.85; Cl, 7.42; N, 18.18; Found:

C, 41.70; H, 3.67; Cl, 7.44; N, 18.08.

Scheme 351

600

CF₃ 346f 351a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyI-6-(trifluoromethyl)-9Hpyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yI)acetyl)-N-(6-chloropyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (351a)

Compound 351a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetic acid (346f) (55 mg, 0.125 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-chloropyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (269b) (36.1 mg, 0.125 mmol), HATU (57.1 mg, 0.150 mmol), DIPEA (81 mg, 0.626 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scherne299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-chloropyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (351a) (56 mg, 80% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-7₆) δ 10.82 (s, 1H, D₂O exchangeable), 8.97 (s, 1H), 8.81 (bs, 2H, D₂O exchangeable), 8.67 (s, 1H), 7.99 (d, J = 8.2 Hz, 1H), 7.81 (t, J = 8.0 Hz, 1H), 7.18 (d, J = 7.6 Hz, 1H), 5.96 (d, J = 18.1 Hz, 1H), 5.68 (d, J = 17.9 Hz, 1H), 4.39 (dd, J = 9.1, 6.2 Hz, 1H), 3.78 - 3.69 (m, 1H), 2.93 (s, 3H), 2.49 - 2.43 (m, 1H), 1.99 (dd, J= 13.3, 6.1 Hz, 1H), 1.31 (s, 3H), 1.08 - 0.98 (m, 1H), 0.92 - 0.81 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-tfc) δ -64.37; MS (ES+): 559/561 (M+l); (ES-): 557/559 (M-l); Analysis calculated for

C₂₅H₂₂ClF₃N₈O₂.0.95HC1.2H₂O: C, 47.69; H, 4.31; Cl, 10.98; N, 17.80; Found: C, 47.78; H,

4.27; Cl, 10.90; N, 17.77.

Scheme 352

601

352c

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(furan-3-yl)-8-methyl-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (352c) Step-1: Préparation of Zeri-butyl 2-(4-amino-6-(furan-3-yl)-8-methyl-9H-pyrimido[4,5b]indol-9-yl)acetate (352a)

Compound 352a was prepared according to the procedure reported in step-lof scheme-62, from tert-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (192e) (200 mg, 0.511 mmol) in dioxane (4 mL) using furan-3-ylboronic acid (57.2 mg, 0.511 mmol), bis(triphenylphosphine)palladium(II) chloride (35.9 mg, 0.051 mmol), a solution of 3.3 M potassium carbonate (0.465 mL, 1.533 mmol) and heating at 100 °C for 16 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with MeOH in DCM from 0-3%] tert-butyl 2-(4-amino-6-(furan-3-yl)-8-methyl-9HpyrimidoÎ4,5-b]indol-9-yl)acetate (352a) (74 mg, 38% yield) as a yellow solid; 'H NMR (300 MHz, DMSO-Jé) δ 8.35 (s, 1H), 8.30 (s, 1H), 8.23 (s, 1H), 7.76 (t, J= 1.7 Hz, 1H), 7.47 (s, 1H), 7.40 (bs, 2H), 7.19 (d, J= 1.9 Hz, 1H), 5.33 (s, 2H), 2.64 (s, 3H), 1.43 (s, 9H); MS (ES+): 379 (M+l); (ES-): 377 (M-l).

Step-2: Préparation of 2-(4-amino-6-(furan-3-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetic acid (352b)

Compound 352b Was prepared according to the procedure reported in step-2 of scheme-1, from terZ-butyl 2-(4-amino-6-(furan-3-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate

602 (352a) (72 mg, 0.190 mmol) in DCM (5 mL) using TFA (217 mg, 1.903 mmol) to afford TFA sait of 2-(4-amino-6-(furan-3-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (352b) (105 mg) as a brown solid; MS (ES+): 323 (M+l); (ES-): 321 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(furan-3-yl)-8-methyl-9H-pyrimido[4,55 b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (352c)

Compound 352c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(furan-3-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (352b) (50 mg, 0.115 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(610 bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (36.5 mg, 0.115 mmol), HATU (52.3 mg, 0.138 mmol), DIPEA (74.0 mg, 0.573 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4amino-6-(furan-3-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (352c) (34 mg, 51% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-ifc) δ 10.84 (s, 1H, D₂O exchangeable), 8.74 (bs, 2H, D₂O exchangeable), 8.61 (s, 1H), 8.52 (s, 1H), 8.27 (s, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.78 (t, J = 1.7 Hz, 1H), 7.71 (t, J =8.0 Hz, 1H), 7.60 (s, 1H),7.31 (d, J = 7.7 Hz, 1H), 7.19 (d, J =1.9 Hz, 1H), 5.91 (d, J = 17.9 Hz, 1H), 5.63 (d, J = 17.9 Hz, 1H), 4.43 (dd, J = 9.1, 5.7 Hz, 1H), 3.98 - 3.89 (m, 1H), 2.73 (s, 3H), 2.42 - 2.29 (m, 1H), 2.30 - 2.12 (m, 1H), 2.01 - 1.84 (m,

1H), 1.22 - 0.95 (m, 1H), 0.77 - 0.60 (m, 1H); MS (ES+): 586/588 (M+l), (ES-): 584/586 (M-l); Analysis calculated for C₂8H₂₄BrN₇O₃.HC1.2.75H₂O: C, 50.01; H, 4.57; Cl, 5.27; N, 14.58; Found: C, 49.94; H, 4.36; Cl, 5.41; N, 14.67.

Scheme 353

352b

353a

603

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(furan-3-yl)-8-methyl-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (353a) '

Compound 353a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(furan-3-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (352b) (50 mg, 0.115 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (38.1 mg, 0.115 mmol), HATU (52.3 mg, 0.138 mmol), DIPEA (74.0 mg, 0.573 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2(4-amino-6-(furan-3-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin2-yl)-5-methyl-2-azabicycIo[3.1.0]hexane-3-carboxamide (353a) (37 mg, 54% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-iZe) δ 10.83 (s, 1H, D₂O exchangeable), 8.71 (bs, 2H, D₂O exchangeable), 8.59 (s, 1H), 8.51 (s, 1H), 8.28 (s, 1H), 8.02 (d, J = 8.2 Hz, 1H), 7.81 - 7.76 (m, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.59 (s, 1H), 7.31 (d, J = 7.7 Hz, 1H), 7.19 (d, J = 1.9 Hz, 1H), 5.83 (d, J = 18.0 Hz, 1H), 5.56 (d, J = 17.9 Hz, 1H), 4.39 (dd, J = 9.0, 6.2 Hz, 1H), 3.70 - 3.64 (m, 1H), 2.69 (s, 3H), 2.49 - 2.43 (m, 1H), 1.98 (dd, J = 13.3, 6.0 Hz, 1H), 1.31 (s, 3H), 1.08 - 0.94 (m, 1H), 0.92 - 0.79 (m, 1H); MS (ES+): 600/602 (M+l); (ES-): 598/600 (M-l); Analysis calculated for C₂9H₂₆BrN₇O3.1.05HC1.2.75H₂O: C, 50.61; H, 4.77; Cl, 5.41; N, 14.24; Found: C, 50.69; H, 4.66; Cl, 5.31; N, 14.36.

Scheme 354

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-4-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (354c)

Step-1 : Préparation of Zeri-butyl 2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-4-yl)9H-pyrimido[4,5-b]indol-9-yl)acetate (354a)

Compound 354a was prepared according to the procedure reported in scheme-263, from tertbutyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (192e) (200 mg, 0.511 mmol) in n-BuOH (4 mL) using (2-(trifluoromethyl)pyridin-4-yl)boronic acid (146 mg, 10 0.Ί67 mmol), Pd₂(dba)₃ (46.8 mg, 0.051 mmol), XPhos (48.7 mg, 0.102 mmol), a solution of

1.27 M potassium phosphate (0.805 mL, 1.022 mmol) and heating at 100 °C for 16 h. This gave after workup and purification using flash column chromatography [silica gel (12 g), eluting with MeOH in DCM from 0-7%] ZerZ-butyl 2-(4-amino-8-methyl-6-(2(trifluoromethyl)pyridin-4-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (354a) (169 mg, 72% yield); *H NMR (300 MHz, DMSO-7₆) δ 8.82 (d, J= 5.2 Hz, 1H), 8.76 - 8.66 (m, 1H), 8.42 (d, 7= 1.6 Hz, 1H), 8.35 (s, 1H), 8.26 (dt,7=5.6, 1.2 Hz, 1H), 7.82 (d, 7= 1.7 Hz, 1H), 7.58 (bs, 2H), 5.39 (s, 2H), 2.74 (s, 3H), 1.44 (s, 9H); ¹⁹F NMR (282 MHz, DMSO-7₆) δ -66.17; MS (ES+): 458 (M+l); (ES-): 456 (M-l).

605

Step-2: Préparation of 2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyndin-4-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (354b)

Compound 354b was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-4-yl)-9H-pyrimido[4,5b]indol-9-yl)acetate (354a) (167 mg, 0.365 mmol) in DCM (5 mL) using TFA (416 mg, 3.65 mmol) to afford after workup TFA sait of 2-(4-amino-8-methyl-6-(2(trifluoromethyl)pyridin-4-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (354b) (233 mg) as a pale-yellow solid; Ή NMR (300 MHz, DMSO-î/ô) δ 8.86 (d, J= 5.2 Hz, 1H), 8.81 (d, J= 1.8 Hz, 1H), 8.57 (s, 1H), 8.45 (s, 2H), 8.41 (d, J= 1.7 Hz, 1H), 8.26 (dd, J= 5.2, 1.7 Hz, 1H), 7.93 (d, J= 1.6 Hz, 1H), 5.48 (s, 2H), 2.79 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-î/ô) δ -66.19, -74.61; MS (ES+): 402 (M+l); (ES-): 400 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-4yl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (354c)

Compound 354c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-4-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (354b) (60 mg, 0.116 mmol) in DMF (5 mL) using HCl sait of ( 1 R,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (37.1 mg, 0.116 mmol), HATU (53.1 mg, 0.140 mmol), DIPEA (75 mg, 0.582 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-(trifhioromethyl)pyridin-4-yl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (354c) (52 mg, 67% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-î/ô) δ 10.84 (s, 1H, D₂O exchangeable), 8.85 (d, J = 5.6 Hz, 1H), 8.84 (s, 1H), 8.79 (bs, 2H, D₂O exchangeable), 8.65 (s, 1H), 8.40 (d, J = 1.7 Hz, 1H), 8.26 (dd, J = 5.3, 1.7 Hz, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.93 (s, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.97 (d, J = 18.0 Hz, 1H), 5.68 (d, J = 17.9 Hz, 1H), 4.44 (dd, J = 9.0, 5.7 Hz, 1H), 4.00 - 3.92 (m, 1H), 2.81 (s, 3H), 2.43 - 2.30 (m, 1H), 2.30 - 2.11 (m, 1H), 2.05 - 1.85 (m, 1H), 1.18 - 1.00 (m, 1H), 0.81 - 0.60 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î/ô) δ -66.18; MS (ES+): 665/667 (M+l), (ES-): 663/665 (M-l); Analysis calculated for C3oH₂4BrF3Ns0₂.1.3HC1.1.75H₂0: C, 48.40; H, 3.90; Cl, 6.19; N, 15.05; Found: C, 48.48; H, 3.70; Cl, 6.30; N, 14.97.

Scheme 355

606

354b 355a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-(trifluoromethyI)pyridin-4-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (355a)

Compound 355a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-4-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (354b) (60 mg, 0.116 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (38.7 mg, 0.116 mmol), HATU (53.1 mg, 0.140 mmol), DIPEA (75 mg, 0.582 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-4-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyI-2azabicyclo[3.1.0]hexane-3-carboxamide (355a) (55 mg, 70% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-cfc) δ 10.84 (s, 1H, D₂O exchangeable), 8.99 - 8.69 (m, 4H, in which 2H were D₂O exchangeable), 8.65 (s, 1H), 8.39 (d, J = 1.7 Hz, 1H), 8.26 (dd, J = 5.2, 1.7 Hz, 1H), 8.02 (d, J = 8.2 Hz, 1H), 7.92 (d, J = 1.7 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.91 (d, J = 18.0 Hz, 1H), 5.63 (d, J = 17.9 Hz, 1H), 4.40 (dd, J = 9.0, 6.1 Hz, 1H), 3.74 - 3.69 (m, 1H), 2.79 (s, 3H), 2.49 - 2.44 (m, 1H), 1.99 (dd, J = 13.2, 6.0 Hz, 1H), 1.32 (s, 3H), 1.08 - 0.96 (m, 1H), 0.93 - 0.81 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ

-66.18; MS (ES+): 679/681 (M+l); (ES-): 677/679 (M-l); Analysis calculated for

C₃iH₂₆BrF₃N₈O₂.0.25HC1.4H₂O: C, 48.95; H, 4.54; Cl, 1.17; N, 14.73; Found: C, 48.94; H, 3.99; Cl, 1.07; N, 14.73.

Scheme 356

607

Β(ΟΗ)₂

cf₃

192e 356a

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(6-(trifluoromethyl)pyridin-3-yI)-9Hpyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (356c)

Step-1 : Préparation of tert-butyl 2-(4-amino-8-methyl-6-(6-(trifluoromethyl)pyridin-3-yl)9H-pyrimido[4,5-b]indol-9-yl)acetate (356a)

Compound 356a was prepared according to the procedure reported in scheme-263 and step-1 of scheme-354, from tert-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetate (192e) (200 mg, 0.511 mmol) and (6-(trifluoromethyl)pyridin-3-yl)boronic acid (146 mg, 0.767 mmol) to afford tert-butyl 2-(4-amino-8-methyl-6-(6(trifhioromethyl)pyridin-3-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (356a) (214 mg, 92% yield) as a white solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 9.30 (d, J= 2.4 Hz, 1H), 8.66 (s, 1H), 8.54 (d, J= 8.3 Hz, 1H), 8.34 (d, J= 1.8 Hz, 1H), 8.01 (dd, J= 8.3, 1.8 Hz, 1H), 7.70 (s, 1H), 7.52 (bs, 2H), 5.38 (s, 2H), 2.73 (s, 3H), 1.45 (s, 9H); ¹⁹F NMR (282 MHz, DMSO d₆) δ

-66.02; MS (ES+): 458 (M+l); (ES-): 456 (M-l).

Step-2: Préparation of 2-(4-amino-8-methyl-6-(6-(trifluoromethyl)pyridin-3-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (356b)

Compound 356b was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-8-methyl-6-(6-(trifluoromethyl)pyridin-3-yl)-9H-pyrimido[4,5

608

b]indol-9-yl)acetate (356a) (210 mg, 0.459 mmol) m DCM (5 mL) using TFA (523 mg, 4.59 mmol) to afford TFA sait of 2-(4-amino-8-methyl-6-(6-(trifluoromethyl)pyridin-3-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (356b) (298 mg) as apale-yellow solid; ’HNMR (300 MHz, DMSO-J₆) δ 9.30 (d, J= 2.2 Hz, 1H), 8.77 (d, J= 1.8 Hz, 1H), 8.58 (s, 1H), 8.55 (dd, J = 8.4, 2.2 Hz, 1H), 8.49 (s, 1H), 8.05 (d, J= 8.3 Hz, 1H), 7.82 (d, J= 1.7 Hz, 1H), 5.48 (s, 2H), 2.78 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-ί/φ) δ -66.07, -74.59; MS (ES+): 402 (M+l); (ES-): 400 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(6-(trifluoromethyl)pyridin-3yl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (356c)

Compound 356c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(6-(trifluoromethyl)pyridin-3-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (356b) (60 mg, 0.116 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (37.1 mg, 0.116 mmol), HATU (53.1 mg, 0.140 mmol), DIPEA (75 mg, 0.582 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(6-(trifluoromethyl)pyridin-3-yl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (356c) (47 mg, 61% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-cfe) δ 10.84 (s, 1H, D₂O exchangeable), 9.29 (d, J = 2.2 Hz, 1H), 8.78 (d, J = 1.8 Hz, 1H), 8.68 (bs, 2H, D₂O exchangeable), 8.62 (s, 1H), 8.54 (dd, J = 8.1, 2.2 Hz, 1H), 8.10 - 7.97 (m, 2H), 7.80 (s, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 5.96 (d, J = 18.0 Hz, 1H), 5.68 (d, J = 17.9 Hz, 1H), 4.44 (dd, J = 9.0, 5.7 Hz, 1H), 4.01 - 3.90 (m, 1H), 2.80 (s, 3H), 2.44 - 2.30 (m, 1H), 2.30 - 2.13 (m, 1H), 2.03 - 1.86 (m, 1H), 1.18 - 1.04 (m, 1H), 0.80 - 0.63 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-ùfe) δ -66.08; MS (ES+): 665/667 (M+l); (ES-): 663/665 (M-l); Analysis calculated for C3oH₂₄BrF₃N₈0₂.l.lHC1.2.5H₂0: C, 48.00; H, 4.04; Cl, 5.20; N, 14.93; Found: C, 48.10; H, 3.54; Cl, 5.27; N, 14.93.

Scheme 357

609

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(6-(trifluoromethyl)pyridin-3-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (357a)

Compound 357a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyI-6-(6-(trifluoromethyl)pyridin-3-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (356b) (60 mg, 0.116 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (38.7 mg, 0.116 mmol), HATU (53.1 mg, 0.140 mmol), DIPEA (75 mg, 0.582 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(6-(trifluoromethyl)pyridin-3-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (357a) (50 mg, 63% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-<&) δ 10.84 (s, 1H, D₂O exchangeable), 9.29 (d, J = 2.2 Hz, 1H), 15 8.99 - 8.69 (m, 3H, in which 2H were D₂O exchangeable), 8.63 (s, 1H), 8.54 (dd, J = 8.0, 2.3

Hz, 1H), 8.04 (d, J = 3.0 Hz, 1H), 8.01 (d, J = 3.0 Hz, 1H), 7.79 (s, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 5.90 (d, J =18.0 Hz, 1H), 5.61 (d, J =17.8 Hz, 1H), 4.40 (dd, J = 9.0, 6.2 Hz, 1H), 3.75 - 3.68 (m, 1H), 2.77 (s, 3H), 2.49 - 2.39 (m, 1H), 1.99 (dd, J = 13.3, 6.0 Hz, 1H), 1.32 (s, 3H), 1.07 - 0.96 (m, 1H), 0.90 - 0.74 (m, 1H); ¹⁹F NMR (282 MHz,

DMSO-cZe) δ -66.09; MS (ES+): 679/681 (M+l); (ES-): 677/679 (M-l); Analysis calculated for C3iH₂₆BrF₃N₈O₂.1.15HC1.2H₂O: C, 49.16; H, 4.15; Cl, 5.38; N, 14.79; Found: C, 49.2.5; H, 3.91; Cl, 5.37; N, 14.83.

Scheme 358 ι

610

192e 358a

358b

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-3-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (358c)

Step-1 : Préparation of ZerZ-butyl 2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-3-yl)9H-pyrimido[4,5-b]indoI-9-yl)acetate (358a)

Compound 358a was prepared according to the procedure reported in scheme-263 and step-1 of scheme-354, from ZerZ-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetate (192e) (200 mg, 0.511 mmol) and (2-(trifluoromethyl)pyridin-3-yl)boronic acid (98 mg, 0.511 mmol) to afford ZerZ-butyl 2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin3-yI)-9H-pyrimido[4,5-b]indol-9-yl)acetate (358a) (21 mg, 9% yield); MS (ES+): 458 (M+l); (ES-): 456 (M-l).

Step-2: Préparation of 2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-3-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (358b)

Compound 358b was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-3-yl)-9H-pyrimido[4,5b]indol-9-yl)acetate (358a) (21 mg, 0.456 mmol) in DCM (5 mL) using TFA (52.3 mg, 0.459 mmol) to afford TFA sait of 2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-3-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (358b) (26 mg) as a pale-yellow solid; MS (ES+): 402 (M+l); (ES-): 400 (M-l).

611

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-3yl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (358c)

Compound 358c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-3-yl)-9H-pyrimido[4,5b]indol-9-yl)acetic acid (358b) (26 mg, 0.050 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (16.78 mg, 0.050 mmol), HATU (23.02 mg, 0.061 mmol), DIPEA (32.6 mg, 0.252 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(2-(trifluoromethyl)pyridin-3-yl)-9Hpyrimido[4,5-b] indol-9-y l)acetyl)-N-(6-bromopyridin-2-yl)-5 -methyI-2azabicyclo[3.1.0]hexane-3-carboxamide (358c) (21 mg, 0.031 mmol, 61.3 % yield) HCl sait as a pale-yellow solid; ’H NMR (300 MHz, DMSO-i/d) δ 10.85 (s, 1H, D2O exchangeable), 8.81 (d, J = 4.7 Hz, 1H), 8.61 (d, J = 2.0 Hz, 1H), 8.42 (bs, 2H, D₂O exchangeable), 8.38 (s,

1H), 8.12 - 7.95 (m, 2H), 7.83 (dd, J = 7.8, 4.7 Hz, 1H), 7.71 (t, J = 7.9 Hz, 1H), 7.32 (d, J =

7.8 Hz, 1H), 7.26 (s, 1H), 5.92 (d, J = 17.9 Hz, 1H), 5.65 (d, J = 18.0 Hz, 1H), 4.49 - 4.33 (m, 1H), 3.74 - 3.69 (m, 1H), 2.73 (s, 3H), 2.49 - 2.45 (m, 1H), 1.99 (dd, J = 13.2, 6.1 Hz, 1H), 1.32 (s, 3H), 1.10 - 0.97 (m, 1H), 0.93 - 0.80 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-îZ₆) δ 59.64; MS (ES+): 679/681 (M+l); (ES-): 677/679 (M-l).

Scheme 359 <

192e 359a

O,

359b

359c

612

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(furan-2-yl)-8-methyl-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyI-2-azabicyclo[3.1.0]hexane-3-carboxamide (359c)

Step-1: Préparation of tert-butyl 2-(4-amino-6-(furan-2-yl)-8-methyl-9H-pyrimido[4,5b]indol-9-yl)acetate (359a)

Compound 359a was prepared according to the procedure reported in scheme-263 and step-1 of scheme-354, from tert-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetate (192e) (180 mg, 0.460 mmol) and furan-2-ylboronic acid (103 mg, 0.920 mmol), to afford tert-butyl 2-(4-amino-6-(furan-2-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (359a) (158 mg, 91% yield)as a pale-yellow solid; *H NMR (300 MHz, DMSO-ifc) δ 8.49 (d, J= 1.7 Hz, 1H), 8.31 (s, 1H), 7.74 (d, J= 1.8 Hz, 1H), 7.54 (s, 1H), 7.42 (bs, 2H), 7.03 (d, J = 3.3 Hz, 1H), 6.62 (dd, J= 3.4, 1.8 Hz, 1H), 5.34 (s, 2H), 2.67 (s, 3H), 1.43 (s, 9H); MS (ES+): 379 (M+l); (ES-): 377 (M-l).

Step-2: Préparation of 2-(4-amino-6-(furan-2-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetic acid (359b)

Compound 359b was prepared according to the procedure reported in step-2 of scheme-1, from terf-butyl 2-(4-amino-6-(furan-2-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (359a) (158 mg, 0.418 mmol) in DCM (5 mL) using TFA (476 mg, 4.18 mmol) to afford TFA sait of 2-(4-amino-6-(furan-2-yI)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (359b) (196 mg) as a green solid; *H NMR (300 MHz, DMSO-cfc) δ 8.54 (d, J= 1.6 Hz, 1H), 8.52 - 8.45 (m, 3H), 7.70 (d, J= 1.8 Hz, 1H), 7.58 (s, 1H), 6.96 (d, J= 3.3 Hz, 1H), 6.57 (dd, J= 3.4, 1.8 Hz, 1H), 5.36 (s, 2H), 2.64 (s, 3H); MS (ES+): 323 (M+l), (ES-): 321 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(furan-2-yl)-8-methyl-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (359c)

Compound 359c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(furan-2-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (359b) (60 mg, 0.138 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (45.7 mg, 0.138 mmol), HATU (62.7 mg, 0.165 mmol), DIPEA (89 mg, 0.688 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2(4-amino-6-(furan-2-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin

613

2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (359c) (27 mg, 33% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-ifc) δ 10.83 (s, 1H, D₂O exchangeable), 8.60 (s, 1H), 8.55 (s, 1H), 8.46 (bs, 2H, D₂O exchangeable), 8.02 (d, J = 8.2 Hz, 1H), 7.77 (d, J = 1.7 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.63 (s, 1H), 7.31 (d, J = 7.7 Hz, 1H), 7.03 (d, J = 3.4 Hz, 1H), 6.69 - 6.59 (m, 1H), 5.86 (d, J = 17.8 Hz, 1H), 5.59 (d, J = 17.8 Hz, 1H), 4.39 (dd, J = 9.1, 6.2 Hz, 1H), 3.73 - 3.67 (m, 1H), 2.72 (s, 3H), 2.49 - 2.42 (m, 1H), 1.99 (dd, J= 13.3, 6.1 Hz, 1H), 1.31 (s, 3H), 1.09 - 0.96 (m, 1H), 0.91 - 0.73 (m, 1H); MS (ES+): 600/602 (M+l); (ES-): 598/600 (M-l).

Scheme 360

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (360c) Step-1: Préparation of tert-butyl 2-(4-amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5b]indol-9-yl)acetate (360a)

Compound 360a was prepared according to the procedure reported in step-1 of scheme-59 and step-1 of scheme-354, from tert-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5b]indol-9-yl)acetate (192e) (180 mg, 0.460 mmol) and cyclopropyl boronic acid (79 mg, 0.920 mmol) to afford tert-butyl 2-(4-amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5b]indol-9-yl)acetate (360a) (157 mg, 97% yield); Ή NMR (300 MHz, DMSO-J₆) δ 8.80 (bs, 2H), 8.63 (s, 1H), 8.00 (d, J= 1.7 Hz, 1H), 7.11 (d, J= 1.6 Hz, 1H), 5.41 (s, 2H), 2.63 (s, 3H), 2.13 - 1.91 (m, 1H), 1.44 (s, 9H), 1.07 - 0.95 (m, 2H), 0.89 - 0.73 (m, 2H); MS (ES+): 353 (M+l); (ES-): 351 (M-l).

614

Step-2: Préparation of 2-(4-amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5-b]indol-9yl)acetic acid (360b)

Compound 360b was prepared according to the procedure reported in step-2 of scheme-1, from terributyl 2-(4-amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (360a) (151 mg, 0.428 mmol) in DCM (5 mL) using TFA (489 mg, 4.28 mmol) to afford

TFA sait of 2-(4-amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (360b) (186 mg) as a white solid; ’H NMR (300 MHz, DMSO-ô/₆) δ 8.63 (bs, 2H), 8.59 (s, 1H), 7.96 (d, J= 1.7 Hz, 1H), 7.10 (s, 1H), 5.41 (s, 2H), 2.65 (s, 3H), 2.13 - 1.94 (m, 1H), 1.09 - 0.93 (m, 2H), 0.88 - 0.72 (m, 2H); MS (ES+): 297 (M+l); (ES-): 295 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (360c)

Compound 360c was prepared according to the procedure reported iri step-3 of scheme-1, from TFA sait of 2-(4-amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic 15 acid (360b) (66 mg, 0.161 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (51.2 mg, 0.161 mmol), HATU (73.4 mg, 0.193 mmol), DIPEA (104 mg, 0.804 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-220 yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (360c) (39 mg, 43% yield) HCl sait as a white solid; ‘H NMR (300 MHz, DMSO-î/₆) δ 10.82 (s, 1H, D₂O exchangeable), 8.89 - 8.33 (m, 3H, in which 2H were D₂O exchangeable), 8.01 (d, J = 8.2 Hz, 1H), 7.96 (d, J = 1.7 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 7.06 (s, 1H), 5.88 (d, J = 17.9 Hz, 1H), 5.61 (d, J = 17.9 Hz, 1H), 4.42 (dd, J = 9.1, 5.7 Hz, 1H), 3.98 - 3.86 (m, 1H), 2.66 (s, 3H), 2.42 25 2.28 (m, 1H), 2.27 - 2.13 (m, 1H), 2.08 - 1.97 (m, 1H), 1.96 - 1.86 (m, 1H), 1.14 - 1.02 (m,

1H), 1.02 - 0.92 (m, 2H), 0.87 - 0.77 (m, 2H), 0.73 - 0.64 (m, 1H); MS (ES+): 560/562 (M+l); (ES-): 558/560 (M-l); Analysis calculated for C₂₇H₂₆BrN₇O₂.1.35HC1.2.75H₂O: C, 49.19; H, 5.02; Cl, 7.26; N, 14.87; Found: C, 49.09; H, 4.77; Cl, 7.30; N, 14.93.

Scheme 361

615

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5-b]indoI9-yl)acetyI)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (361a)

Compound 361a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (360b) (60 mg, 0.146 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (48.6 mg, 0.146 mmol), HATU (66.7 mg, 0.175 mmol), DIPEA (94 mg, 0.731 mmol) and stirring at RT for

16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2(4-amino-6-cyclopropyl-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (361a) (49 mg, 58% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-i/d) δ 10.82 (s, 1H, D₂O exchangeable), 8.73 (bs, 2H, D₂O exchangeable), 8.61 (s, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.96 (d, J = 1.7 Hz, 1H),

7.71 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 7.7 Hz, 1H), 7.06 (d, J = 1.6 Hz, 1H), 5.83 (d, J = 17.9

Hz, 1H), 5.56 (d, J = 17.9 Hz, 1H), 4.38 (dd, J = 9.0, 6.1 Hz, 1H), 3.72 - 3.66 (m, 1H), 2.64 (s, 3H), 2.49 - 2.36 (m, 1H), 2.07 - 1.89 (m, 2H), 1.31 (s, 3H), 1.06 - 0.91 (m, 3H), 0.89 0.76 (m, 3H); MS (ES+): 574/576 (M+l); (ES-): 572/574 (M-l); Analysis calculated for C₂8H₂8BrN₇O₂.HC1.2H₂O: C, 51.98; H, 5.14; Cl, 5.48; N, 15.15; Found: C, 51.82; H, 5.01;

Cl, 5.44; N, 15.22.

Scheme 362

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyI)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (362f)

Step-1: Préparation of 2-amino-7-methyl-5-(trifluoromethyl)-lH-pyrrolo[3,2-b]pyridine-3carbonitrile (362b)

Compound 362b was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-4-methyl-6-(trifluoromethyl)pyridin-3-yl)-2,2,2-trifluoroaçetamide (362a) (4.87 g, 13.87 mmol; CAS # 2055497-18-0) in DMSO (30 mL) using malononitrile (1.100 g,

16.65 mmol), L-proline (0.319 g, 2.77 mmol), Cul (0.264 g, 1.387 mmol), a solution of

K2CO3 (3.83 g, 27.7 mmol) in water (10 mL) to afford after purification [S1O2 gel (80 g), MeOH in DCM from 0-10%] 2-amino-7-methyl-5-(trifluoromethyl)-lH-pyrrolo[3,2b]pyridine-3-carbonitrile (362b) (3.38 g) as apale-yellow solid; *HNMR (300 MHz, DMSOJ₆) δ 11.52 (s, 1H), 7.31 (s, 2H), 7.22 (s, 1H), 2.43 (s, 3H); ¹⁹F NMR (282 MHz, DMSO) δ 15 64.38; MS (ES+): 241 (M+l); (ES-): 239 (M-l).

Step-2: Préparation of 8-methyl-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-4-amine (362c)

Compound 362c was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-7-methyl-5-(trifluoromethyl)-lH-pyrrolo[3,2-b]pyridine-3-carbonitrile (362b) (3.38 g, 14.07 mmol) using trimethoxymethane (7.47 g, 70.4 mmol) and NH₄OAc (3.25 g,

42.2 mmol) to afford 8-methyl-6-(trifluoromethyl)-9H-pyrido[2^,,3':4,5]pyrrolo[2,3d]pyrimidin-4-amine (362c) (3.28 g, 87% yield) as a green solid; Ή NMR (300 MHz,

617

DMSO-ôfc) δ 12.57 (s, 1H), 8.43 (s, 1H), 7.99 (s, 1H), 7.72 (s, 1H), 6.46 (s, 1H), 2.66 (s, 3H); ¹⁹F NMR (282 MHz, DMSOX) δ -64.00; MS (ES+): 268 (M+l); (ES-): 266 (M-l).

Step-3 : Préparation of ZerZ-butyl 2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (362d)

Compound 362d was prepared according to the procedure reported in step-1 of scheme-1, from 8-methyl-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-4-amine (362c) (3.28 g, 12.27 mmol) in DMF (15 mL) using ZerZ-butyl 2-bromoacetate (2.63 g, 13.50 mmol) and CS2CO3 (8.0 g, 24.55 mmol) to afford Zeri-butyl 2-(4-amino-8-methyl-6(trifluoromethyI)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetate (362d) (3.21 g, 10 69% yield) as a pale-yellow solid; *H NMR (300 MHz, DMSO-ôfc) δ 8.48 (s, 1H), 8.20 (s,

1H), 7.75 (s, 1H), 6.64 (s, 1H), 5.39 (s, 2H), 2.74 (s, 3H), 1.43 (s, 9H); ¹⁹F NMR (282 MHz, DMSO-î/ô) δ -64.31; MS (ES+) 382 (M+l).

Step-4: Préparation of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (362e)

Compound 362e was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrroIo[2,3d]pyrimidin-9-yl)acetate (362d) (3.21 g, 8.42 mmol) using TFA (9.6 g, 84 mmol) in DCM (10 mL) to afford TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetic acid (362e) (3.16 g, 85% yield) as apale20 yellow solid; Ή NMR (300 MHz, DMSO-7₆) δ 8.57 (s, 2H), 7.80 (s, 1H), 7.09 (bs, 1H), 5.43 (s, 2H), 2.78 (s, 3H); ¹⁹F NMR (282 MHz, DMSO) δ -64.34, -74.87; MS (ES+): 326 (M+l); (ES-): 324 (M-l).

Step-5: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-225 azabicyclo[3.1.0]hexane-3-carboxamide (362f)

Compound 362f was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetic acid (362e) (75 mg, 0.171 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (54.4 mg, 0.171 mmol), HATU (78 mg, 0.205 mmol), DIPEA (110 mg, 0.854 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3

618

d]pyrimidin-9-yl)acetyI)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (362f) (76 mg, 76% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-cfc) δ 10.77 (s, 1H, D₂O exchangeable), 8.52 (bs, 2H, in which 1H was D₂O exchangeable), 7.93 (d, J = 8.2 Hz, 1H), 7.71 (s, 1H), 7.64 (t, J = 8.0 Hz, 1H), 7.25 (d, J = 7.7 Hz, 1H), 7.13 (bs, 1H, D₂O exchangeable), 5.84 (d, J = 18.0 Hz, 1H), 5.59 (d, J = 17.9 Hz, 1H), 4.36 (dd, J = 9.1, 5.7 Hz, 1H), 3.91 - 3.83 (m, 1H), 2.72 (s, 3H), 2.36 - 2.22 (m, 1H), 2.19 - 2.08 (m, 1H), 1.93 - 1.77 (m, 1H), 1.09 - 0.94 (m, 1H), 0.71 - 0.58 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-cfe) δ -64.32; MS (ES+): 589/591 (M+l); (ES-): 587/589 (M-l).

Scheme 363

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (363a)

Compound 363a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetic acid (362e) (75 mg, 0.171 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (56.8 mg, 0.171 mmol), HATU (78 mg, 0.205 mmol), DIPEA (110 mg, 0.854 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (363a) (71 mg, 69% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-t/e) δ 10.84 (s, 1H, D₂O exchangeable), 8.82 (bs, 1H, D₂O exchangeable), 8.63 (s, 1H), 8.01 (d, J = 8.1 Hz, 1H), 7.80 (s, 1H), 7.71 (t, J = 7.9 Hz, 1H), 7.41 (bs, 1H, D₂O exchangeable), 7.32 (d, J = 7.7 Hz, 1H), 5.88 (d, J = 18.0 Hz, 1H), 5.62 (d, J = 17.9 Hz, 1H), 4.39 (dd, J = 9.1, 6.1 Hz, 1H), 3.76 - 3.67 (m, 1H), 2.78 (s, 3H), 2.50 - 2.38 (m, 1H), 1.98

619 (dd, J= 13.3, 6.0 Hz, 1H), 1.31 (s, 3H), 1.08 - 0.98 (m, 1H), 0.93 - 0.78 (m, 1H); ¹⁹FNMR (282 MHz, DMSO-Jô) δ -64.34; MS (ES+): 603/605 (M+l); (ES-): 601/603 (M-l).

Scheme 364

364d 364e

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloro-5-methylpyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane3-carboxamide (364e) and (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-5-methylpyrazin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (364d)

Step-1: Préparation of (lR,3S,5R)-tert-butyl 3-((6-bromo-5-methylpyrazin-2-yl)carbamoyl)5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (364a)

Compound 364a was prepared according to the procedure reported in step-1 of scheme-53 and step-1 of scheme-317, from (lR,3S,5R)-2-(tert-butoxycarbonyl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxylic acid (261a) (0.321 g, 1.330 mmol) and 6-bromo-5methylpyrazin-2-amine (0.25 g, 1.330 mmol) to afford (lR,3S,5R)-tert-butyl 3-((6-bromo-5methylpyrazin-2-yl)carbamoyl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (364a) (0.36 g, 66% yield) as a white crystalline solid; ’H NMR (300 MHz, DMSO-î/ô) δ 11.13 — 10.84 (m, 1H), 9.33 - 9.08 (m, 1H), 4.23-4.03 (m, 1H), 3.24 - 3.09 (m, 1H), 2.56 (s, 3H), 2.40 (dd, J= 12.9, 8.8 Hz, 1H), 1.98 - 1.77 (m, 1H), 1.32 (d, 9H), 1.19 (s, 3H), 0.75 - 0.55 (m, 2H); MS (ES+): 411/413 (M+l); (ES-): 409/411 (M-l).

Step-2: Préparation of (lR,3S,5R)-N-(6-bromo-5-methylpyrazin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (364b) and (lR,3S,5R)-N-(6-chloro-5methylpyrazin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (364c)

620

Compounds 364b and 364c were prepared according to the procedure reported in step-2 of scheme-344, from (lR,3S,5R)-tert-butyl 3-((6-bromo-5-methylpyrazin-2-yI)carbamoyI)-5methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (364a) (360 mg, 0.875 mmol) in DCM (5 mL) using 4 M HCl in dioxane (2.188 mL, 8.75 mmol) to afford 318 mg HCl sait of a mixture oftwo products (lR,3S,5R)-N-(6-bromo-5-methylpyrazin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (364b); MS (ES+): 311/313 (M+l); (ES-): 309/311 (M-l), and (lR,3S,5R)-N-(6-chloro-5-methylpyrazin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (364c); MS (ES+): 267/269 (M+l); (ES-): 265/267 (M-l) as white solids.

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloro-5-methylpyrazin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (364e) and (lR,3S,5R)-2-(2-(4-amino-8-methyl-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-5-methylpyrazin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (364d)

Compounds 364d and 364e were prepared according to the procedure reported in step-3 of scheme-1, from above mixture of (lR,3S,5R)-N-(6-bromo-5-methylpyrazin-2-yl)-5-methyl2-azabicyclo[3.1.0]hexane-3-carboxamide (364b) and (lR,3S,5R)-N-(6-chloro-5methylpyrazin-2-yI)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (364c) (99 mg, 0.285 mmol) in DMF (5 mL) using HCl sait of 2-(4-amino-8-methyl-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetic acid (1841) (125 mg, 0.285 mmol), HATU (163 mg, 0.428 mmol), DIPEA (184 mg, 1.426 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-methyl-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloro-5-methylpyrazin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (364e) (10 mg, 6% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 11.05 (s, 1H, D₂O exchangeable), 9.10 (s, 1H), 8.79 (s, 1H), 8.65 (bs, 2H, D₂O exchangeable), 8.63 (s, 1H), 7.60 (d, J = 1.7 Hz, 1H), 5.93 (d, J = 18.0 Hz, 1H), 5.65 (d, J = 18.0 Hz, 1H), 4.40 (dd, J = 9.1, 6.1 Hz, 1H), 3.74 - 3.69 (m, 1H), 2.76 (s, 3H), 2.51 (s, 3H), 2.49 - 2.44 (m, 1H), 2.01 (dd, J= 13.3, 6.0 Hz, 1H), 1.32 (s, 3H), 1.08 - 0.97 (m, 1H), 0.94 - 0.81 (m, 1H); ^I9F NMR (282 MHz, DMSO-7₆) δ -58.76; MS (ES+): 573/575 (M+l); (ES-): 571/573 (M-l); and (lR,3S,5R)-2-(2-(4-amino-8-methyl-6(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-5-methylpyrazin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (364d) (77 mg, 44% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-7₆) δ 11.08 (s, 1H, D₂O exchangeable), 9.11 (s, 1H), 8.80 (bs, 3H, in which 2H were D₂O exchangeable), 8.66 (s, 1H), 7.62 (s, 1H), 5.94 (d, J =

621

18.0 Hz, 1H), 5.66 (d, J = 18.0 Hz, 1H), 4.40 (dd, J = 9.1, 6.1 Hz, 1H), 3.75 - 3.69 (m, 1H),

2.76 (s, 3H), 2.52 (s, 3H), 2.50 - 2.45 (m, 1H), 2.01 (dd, J = 13.2, 6.0 Hz, 1H), 1.32 (s, 3H), 1.11- 0.98 (m, 1H), 0.98 - 0.78 (m, 1H); ¹⁹F NMR (282 MHz, DMSOW) δ -58.80; MS (ES+): 617/619 (M+l); (ES-): 615/617 (M-l); Analysis calculated for C26H24BrF3N₈O2.1.25HC1.2.5H₂O: C, 44.10; H, 4.31; Cl, 6.26; N, 15.83; Found: C, 43.91; H, 4.15; Cl, 6.31; N, 15.93

Scheme 365

F

Préparation of (2S,4R)-1 -(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-chloro-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (365c)

Step-1: Préparation of (2S,4R)-/erZ-butyl 2-((6-chloro-3-methylpyridin-2-yl)carbamoyl)-4fluoropyrrolidine-1 -carboxylate (365a)

Compound 365a was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4R)-l-(tert-butoxycarbonyl)-4-fluoropyrrolidine-2-carboxylic acid (321a) (0.340 g, 1.458 mmol) and 6-chloro-3-methylpyridin-2-amine (0.208 g, 1.458 mmol) to afford after purification as reported in step-1 of scheme-317, (2S,4R)-/erZ-butyl 2-((6-chloro-3methyIpyridin-2-yl)carbamoyl)-4-fluoropyrrolidine-l-carboxylate (365a) (0.150 g, 29% yield) as a white solid; MS (ES+): 358.1(M+1); 380.1 (M+Na); (ES-): 356.1(M-1).

Step-2: Préparation of (2S,4R)-N-(6-chloro-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2carboxamide (365b)

622

Compound 365b was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-terZ-butyl 2-((6-chloro-3-methylpyridin-2-yl)carbamoyl)-4-fluoropyrrolidine-lcarboxylate (365a) (0.145 g, 0.405 mmol) in DCM (2.2 mL) using TFA (0.219 mL, 2.84 mmol) to afford after purification as reported in step-2 of scheme-319, HCl sait of (2S,4R)-N(6-chloro-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (365b) (0.086 mg, 82% yield); 'H NMR (300 MHz, DMSO-î/₆) δ 10.96 (s, 1H), 10.22 (s, 1H), 9.05 (s, 1H), 7.83 (d, J = 8.0 Hz, 1H), 7.42 (d, J= 8.0 Hz, 1H), 5.69 - 5.34 (m, 1H), 4.66 (s, 1H), 3.59 (d, ./= 28.5 Hz, 2H), 2.87-2.72 (m, 1H), 2.40-2.22 (m, 1H), 2.18 (s, 3H); MS (ES+): 258.0 (M+l).

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyI)-9H-pyrimido[4,5b]indol-9-yI)acetyl)-N-(6-chIoro-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (365c)

Compound 365c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (2S,4R)-N-(6-chloro-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2carboxamide (365b) (40.3 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of2-(4-amino8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (184f) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (2S,4R)-1-(2-(4amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-chloro-3methylpyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide (365c) (37mg, 48% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) (a mixture of two rotamers) δ 10.99 and 10.52 (2s, 1H, D₂O exchangeable), 8.92 (s, 2H, D₂O exchangeable), 8.83 (s, 1H), 8.68 (s, 1H), 7.83 and 7.69 (2d, J= 8.0 Hz, 1H), 7.64 and 7.61 (2s, 1H), 7.40 and 7.29 (2d, J= 7.9 Hz, 1H), 5.83 (d, J= 18.1 Hz, 1H), 5.69 - 5.41 (m, 2H), 4.63 (t, J= 8.5 Hz, 1H), 4.37 - 4.26 (m, 1H), 4.08-3.92 (m, 1H), 2.74 (s, 3H), 2.66-2.55 (m, 1H), 2.32-2.08 (m, 1H), 1.97 (s, 3H); ¹⁹F NMR (282 MHz, DMSO-J₆) δ -58.82, -176.22; MS (ES+): 564.2 (M+l);586.1 (M+Na); (ES): 562.1 (M-l); Analysis calculated for C₂5H₂₂C1F₄N₇O₂ 2H₂O.0.85HCl: C, 47.59; H, 4.29; Cl, 10.40; N, 15.54; Found: C, 47.87; H, 4.01; Cl, 10.51; N, 15.43.

Scheme 366

623

F F

366c

Préparation of (2S,4R)-1 -(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2carboxamide (366c)

Step-1 : Préparation of (2S,4R)-ZerZ-butyl 2-((6-bromo-5-fluoro-3-methylpyridin-2yl)carbamoyl)-4-fluoropyrrolidine-1 -carboxylate (366a)

Compound 366a was prepared according to the procedure reported in step-1 of scheme-53, from (2S,4R)-l-(ZerZ-butoxycarbonyl)-4-fluoropyrrolidine-2-carboxylic acid (321a) (0.340 g, 1.458 mmol) and 6-bromo-5-fluoro-3-methylpyridin-2-amine (0.299 g, 1.458 mmol) to afford after purification as reported in step-1 of scheme-317, (2S,4R)-ZerZ-butyl 2-((6-bromo-5fluoro-3-methylpyridin-2-yl)carbamoyl)-4-fluoropyrrolidine-l-carboxylate (366a) (0.445 g, 73% yield) as a white solid; MS (ES+): 420.1/422.1 (M+l); 442.0/444.0 (M+Na).

Step-2: Préparation of (2S,4R)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-4fluoropyrrolidine-2-carboxamide (366b)

Compound 366b was prepared according to the procedure reported in step-2 of scheme-1, from (2S,4R)-ZerZ-butyl 2-((6-bromo-5-fluoro-3-methylpyridin-2-yl)carbamoyl)-4fluoropyrrolidine-l-carboxylate (366a) (0.435 g, 1.035 mmol) in DCM (5.6 mL) using TFA (0.558 mL, 7.25 mmol) to afford after purification as reported in step-2 of scheme-319, HCl sait of (2S,4R)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-4-fluoropyrrolidine-220 carboxamide (366b) (0.324 mg, 98% yield); Ή NMR (300 MHz, DMSO-î/ô) δ 10.98 (s, 1H), 10.18 (s, 1H), 9.06 (s, 1H), 7.94 (d, J= 8.4 Hz, 1H), 5.53 (dd, J= 53.5, 3.5 Hz, 1H), 4.64 (t, J = 9.2 Hz, 1H), 3.68-3.60 (m, 1H), 3.57 - 3.51 (m, 1H), 2.88-2.69 (m, 1H), 2.42-2.23 (m, 1H), 2.19 (s, 3H); MS (ES+): 320.0/322.0 (M+l).

624

Step-3: Préparation of (2S,4R)-l-(2-(4-amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2carboxamide (366c)

Compound 366c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of (2S,4R)-N-(6-bromo-5-fluoro-3-methylpyridin-2-yl)-4-fluoropyrrolidine-2carboxamide (366b) (48.8 mg, 0.137 mmol) in DMF (1.5 mL) using TFA sait of 2-(4-amino8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (1841) (60 mg, 0.137 mmol), HATU (78 mg, 0.205 mmol), DIPEA (0.119 mL, 0.684 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (2S,4R)-l-(2-(410 amino-8-methyl-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromo-5fluoro-3-methylpyridin-2-yl)-4-fluoropyrrolidme-2-carboxamide (366c) (63mg, 74% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-ùfe) (a mixture of two rotamers) δ 10.88 and 10.54 (2s, 1H, D2O exchangeable), 8.78 (s, 1H), 8.57 (s, 1H), 8.48 (s, 2H, D2O exchangeable), 7.94 and 7.80 (2d, J= 8.5 Hz, 1H), 7.59 and 7.56 (2s, 1H), 5.80 (d, J= 18.0

Hz, 1H), 5.69 - 5.46 (m, 2H), 4.60 (t, J= 8.5 Hz, 1H), 4.31 (dd, 21.4, 12.7 Hz, 1H), 4.07 - 3.90 (m, 1H), 2.72 (s, 3H), 2.68 - 2.58 (m, 1H), 2.32 - 2.07 (m, 1H), 1.98 (s, 3H). ¹⁹F NMR (282 MHz, DMSO-i/é) δ -58.69, -119.17, -176.67. MS (ES+): 626.1/628.1 (M+l); (ES-): 624.0/626.0 (M-l); Analysis calculated for C25H2iBrF5N?O2 I.5H2O.HCI: C, 43.53; H, 3.65;, Cl, 5.14; N, 14.21; Found: C, 43.65; H, 3.48; Cl, 5.01; N, 14.04.

Scheme 367

625

367e

367f

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methoxy-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (367g)

Step-1: Préparation of N-(2-bromo-5-methoxy-4-(trifluoromethyl)phenyl)-2,2,2trifluoroacetamide (367b)

Compound 367b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-5-methoxy-4-(trifluoromethyl)aniline (367a)(l g, 3.70 mmol; CAS # 69451419-7) in DCM (15 mL) using triethylamine (0.877 mL, 6.30 mmol), trifluoroacetic acid anhydride (0.772 mL, 5.55 mmol) and stirring at RT for 16 h. This gave after workup and purification using flash column chromatography [silica gel (24 g), eluting with EtOAc in hexane from 0 to 50 %] N-(2-bromo-5-methoxy-4-(trifluoromethyl)phenyl)-2,2,2trifluoroacetamide (367b) (1.35 g, 100% yield) as a pale yellow solid; *H NMR (300 MHz, DMSO-î/ô) δ 11.57 (s, 1H), 7.96 (s, 1H), 7.47 (s, 1H), 3.91 (s, 3H); ^I9F NMR (282 MHz,

DMSO-î/6) δ -61.15, -74.02; MS (ES-): 363.9 (M-l).

Step-2: Préparation of 2-amino-6-methoxy-5-(trifluoromethyl)-lH-indole-3-carbonitrile (367c)

626

Compound 367c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-5-methoxy-4-(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (367b) (1.3 g, 3.55 mmol) in DMSO (5 mL) and using malononitrile (0.268 mL, 4.26 mmol), Lproline (0.082 g, 0.710 mmol), Cul (0.068 g, 0.355 mmol) and K2CO3 (0.982 g, 7.10 mmol) 5 to afford after workup and purification as described in step-1 of scheme-328, 2-amino-6methoxy-5-(trifluoromethyl)-lH-indole-3-carbonitrile (367c) (0.106 g, 12% yield) as a yellow solid; Ή NMR (300 MHz, DMSO-J₆) δ 10.98 (s, 1H), 7.31 (s, 1H), 7.07 (s, 1H), 6.99 (s, 2H), 3.89 (s, 3H); ¹⁹F NMR (282 MHz, DMSO) δ -59.05; MS (ES+): 256.1 (M+l); (ES-): 254.0 (M-l).

Step-3: Préparation of 7-methoxy-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-4-amine (367d)

Compound 367d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-6-methoxy-5-(trifluoromethyl)-lH-indole-3-carbonitrile (367c) (0.1 g, 0.392 mmol) in éthanol (10 mL) using formamidine acetate (0.33 g, 3.13 mmol) and refluxing for 75 22 h. The mixture was purified using reverse-phase column chromatography [Cl8 column (50 g), eluting with ACN in water (containing 0.1% HCl) from 0-100%]] to afford 7methoxy-6-(trifluorornethyl)-9H-pyrimido[4,5-b]indol-4-amine (367d) (80 mg, 72% yield) HCl sait as a pale yellow solid; Ή NMR (300 MHz, DMSO-rf₆) δ 13.07 (s, 1H), 8.81 (s, 1H), 8.59 - 8.42 (m, 3H), 7.27 (s, 1H), 3.99 (s, 3H); MS (ES+): 283.1(M+1).

Step-4: Préparation of tert-butyl 2-(4-amino-7-methoxy-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetate (367e)

Compound 367e was prepared according to the procedure reported in step-2 of scheme-16, from HCl sait of 7-methoxy-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-4-amine (367d) (75 mg, 0.235 mmol) in DMF (3 mL) using tert-butyl 2-bromoacetate (0.038 mL, 0.259 25 mmol), CS2CO3 (192 mg, 0.588 mmol) and stirring at RT for 16 h. The solid separated was collected by filtration to give tert-butyl 2-(4-amino-7-methoxy-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetate (367e) (85 mg, 91% yield) as a pale yellow solid; MS (ES+): 397.1 (M+l).

Step-5 : Préparation of 2-(4-amino-7-methoxy-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol30 9-yl)acetic acid (367f)

Compound 367f was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-7-methoxy-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (367e) (83 mg, 0.209 mmol) using 20% TFA in DCM (1202 pL, 3.14 mmol)) to

627 afford TFA sait of 2-(4-ammo-7-methoxy-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (367f) (70 mg, 98% yield) as a pale-yellow solid; 'H NMR (300 MHz, DMSOî/ô) δ 8.75 (s, 1H), 8.45 (s, 1H), 8.01 (d, J= 31.6 Hz, 1H), 7.67 (s, 1H), 5.27 (s, 2H), 3.99 (s, 3H); MS (ES+): 341.1 (M+l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methoxy-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (367g)

Compound 367g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-methoxy-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (367f) (45.4 mg, 0.1 mmol) in DMF (1 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (31.9 mg, 0.100 mmol), HATU (57.0 mg, 0.150 mmol), DIPEA (0.087 mL, 0.500 mmol) and stirring at RT for 1 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2-(2-(4amino-7-methoxy-6-(triflùoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (367g) (48 mg, 79% yield) HCl sait as a white solid; *H NMR (300 MHz, DMSO-î/ô) δ 10.73 (s, 1H, D₂O exchangeable), 8.84 (s, 1H), 8.71 - 8.52 (m, 3H, 2H D₂O exchangeable), 8.02 (d, J= 8.1 Hz, 1H), 7.72 (t, J = 8.0 Hz, 1H), 7.52 (s, 1H), 7.33 (d, J= 7.7 Hz, 1H), 5.77 (d, J= 17.3 Hz, 1H), 5.51 (d, J= 17.2 Hz, 1H), 4.42 (dd, J= 9.1, 5.7 Hz, 1H), 4.01 (s, 3H), 3.93 (td, J= 6.2, 5.3, 2.3 Hz, 1H), 2.36 (dd, J= 13.4, 9.2 Hz, 1H), 2.31 - 2.16 (m, 1H), 2.00-1.88 (m, 1H), 1.17-1.02 (m, 1H), 0.82 - 0.73 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î/ô) δ -59.20; MS (ES+): 604.1/606.1 (M+l); (ES-): 602.0/604.1 (M-l); Analysis calculated for C₂5H₂iBrF3N7O3.3.5H₂O.0.65HCl: C, 43.45; H, 4.18; Cl, 3.33; N, 14.19; Found: C, 43.07; H, 3.76; Cl, 3.04; N, 13.78.

Scheme 368

367f ^368a

628

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-methoxy-6-(trifluoromethyl)-9H-pynmido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (368a)

Compound 368a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-methoxy-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (367f) (45.4 mg, 0.1 mmol) in DMF (1 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (33.3 mg, 0.100 mmol), HATU (57.0 mg, 0.150 mmol), DIPEA (0.087 mL, 0.500 mmol) and stirring at RT for 1 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2(2-(4-amino-7-methoxy-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (368a) (44 mg, 71% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-cfe) δ 10.72 (s, 1H, D₂O exchangeable), 8.85 (s, 1H), 8.70 (s, 2H, D2O exchangeable), 8.61 (s, 1H), 8.02 (d, J= 8.2 Hz, 1H), 7.72 (t, J= 8.0 Hz, 1H), 7.53 (s, 1H), 7.33 (d, J= 7.7 Hz, 1H), 5.74 (d, J= 17.3 Hz,. 1H), 5.46 (d, J= 17.2 Hz, 1H), 4.38 (dd, J= 9.1, 6.1 Hz, 1H), 4.00 (s, 3H), 3.70 (dd, J= 5.5, 2.4 Hz, 1H), 2.57-2.53 (m, 1H), 2.01 (dd, J= 13.3, 6.0 Hz, 1H), 1.33 (s, 3H), 1.09-0.97 (m, 1H), 0.97 - 0.88 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-J₆) δ -59.24; MS (ES+): 618.1/620.1 (M+l); (ES-): 616.1/618.0 (M-l); Analysis calculated for C26H23BrF3N?O3 2H₂O.1.2HC1: C, 44.73; H, 4.07; Cl, 6.09; N, 14.04; Found: C, 44.79; H, 4.07; Cl, 5.87; N, 13.85.

Scheme 369

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-fluoro-6-(trifhioromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (369g)

Step-1: Préparation of N-(2-bromo-6-fluoro-4-(trifluoromethyl)phenyl)-2,2,2trifluoroacetamide (369b)

Compound 369b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-6-fluoro-4-(trifluoromethyl)aniline (369a) (1 g, 3.88 mmol; CAS # 103432563-7) in DCM (15 mL) using triethylamine (0.918 mL, 6.59 mmol), trifluoroacetic acid anhydride (0.808 mL, 5.81 mmol) to afford after purification as reported in step-1 of scheme367 N-(2-bromo-6-fluoro-4-(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (369b) (1.276 g, 93% yield) as a pale yellow solid; ’H NMR (300 MHz, DMSO-î/ô) δ 11.84 (s, 1H), 8.13 (s, 1H), 8.04 (dd, J= 9.4, 1.9 Hz, 1H); MS (ES-): 351.9/353.9 (M-l).

Step-2: Préparation of 2-amino-7-fluoro-5-(trifluoromethyl)-lH-indole-3-carbonitrile (369c) 15 Compound 369c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2-bromo-6-fluoro-4-(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (369b) (1.25 g, 3.53 mmol) in DMSO (5 mL) using malononitrile (0.267 mL, 4.24 mmol), L-proline (0.081 g, 0.706 mmol), Cul (0.067 g, 0.353 mmol), a solution of K2CO3 (0.976 g, 7.06 mmol)

630 in water (5 mL) to afford after purification as reported m step-2 of scheme-367, 2-amino-7fluoro-5-(trifluoromethyl)-lH-indole-3-carbonitrile (369c)(0.125 g, 15% yield) as ayellow solid; ’H NMR (300 MHz, DMSO-îZ₆) δ 11.89 (s, 1H), 7.26 (s, 1H), 7.17 (d, J= 11.0 Hz, 1H), 7.11 (s, 2H); MS (ES+): 244.0 (M+l); (ES-): 242.0 (M-l).

Step-3: Préparation of 8-fluoro-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-4-amine (369d) Compound 369d was prepared according to the procedure reported in step-2 of scheme-29, from 2-amino-7-fluoro-5-(trifluoromethyl)-lH-indole-3-carbonitriIe (369c) (0.120 g, 0.494 mmol) in éthanol (10 mL) using formamidine acetate (0.415 g, 3.95 mmol) to afford after purification as reported in step-3 of scheme-367, 8-fluoro-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-4-amine (369d) (61 mg, 46% yield) HCl sait as a pale yellow solid; *H NMR (300 MHz, DMSO-J₆) δ 13.69 (s, 1H), 8.83 (s, 1H), 8.67 (s, 2H), 8.60 (s, 1H), 7.79 (dd, J=11.0, 1.4 Hz, 1H); MS (ES+): 271.1(M+1).

Step-4: Préparation of terZ-butyl 2-(4-amino-8-fluoro-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetate (369e)

Compound 369e was prepared according to the procedure reported in step-2 of scheme-16, from HCl sait of 8-fluoro-6-(trifhioromethyl)-9H-pyrimido[4,5-b]indol-4-amine (369d) (60 mg, 0.196 mmol) in DMF (3 mL) using ZerZ-butyl 2-bromoacetate (0.032 mL, 0.215 mmol), CS2CO3 (159 mg, 0.489 mmol) and stirring at RT for 16 h. The solid separated was collected by filtration to give fôrf-butyl 2-(4-amino-8-fluoro-6-(trifluoromethyl)-9H-pyrimido[4,5b]indol-9-yl)acetate (369e) (30 mg, 40% yield) as a pale yellow solid; MS (ES+): 385.1(M+1).

Step-5: Préparation of 2-(4-amino-8-fluoro-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (369f)

Compound 369f was prepared according to the procedure reported in step-2 of scheme-1, from terf-butyl 2-(4-amino-8-fluoro-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetate (369e) (28 mg, 0.073 mmol) using 20% TFA in DCM (418 pL, 1.093 mmol)) to afford 2-(4-amino-8-fluoro-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (369f) (23 mg, 96% yield) TFA sait as a pale-yellow solid; 'H NMR (300 MHz, DMSO-cfe) δ 8.73 (s, 1H), 8.41 (s, 1H), 7.80 (s, 2H), 7.70 (d, J= 12.3 Hz, 1H), 5.25 (s, 2H); MS (ES+): 329.0(M+l).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-fluoro-6-(trifluoromethyl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3carboxamide (369g)

631

Compound 369g was prepared according to the procedure reported m step-3 of scheme-1, from TFA sait of 2-(4-amino-8-fhioro-6-(trifhioromethyl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (369f) (31.0 mg, 0.07 mmol) in DMF (1 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (22.30 mg, 0.070 mmol), HATU (39.9 mg, 0.105 mmol), DIPEA (0.061 mL, 0.350 mmol) and stirring at RT for 16 h. This gave after workup and purification as described in scheme-303, (lR,3S,5R)-2(2-(4-amino-8-fluoro-6-(trifluoromethyl)-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (369g) (35 mg, 84 % yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î7₆) δ 10.78 (s, 1H, D₂O exchangeable), 8.81 (s, 1H), 8.60 (s, 1H), 8.48 (s, 2H, D₂O exchangeable), 8.01 (d, J= 8.2 Hz, 1H), 7.81 7.65 (m, 2H), 7.32 (d, J= 7.7 Hz, 1H), 5.90 - 5.73 (m, 1H), 5.50 (dd, J= 17.3, 2.0 Hz, 1H), 4.43 (dd, J= 9.0, 5.6 Hz, 1H), 3.91 - 3.83 (m, 1H), 2.34 (dd, J= 13.4, 9.1 Hz, 1H), 2.30 2.15 (m, 1H), 2.00- 1.86 (m, 1H), 1.16-1.05 (m, 1H), 0.70-0.58 (m, 1H); ¹⁹FNMR(282 MHz, DMSO-riô) δ -58.66, -132.55; MS (ES+): 592.1/594.1 (M+l); (ES-): 590.0/592.0 (ΜΙ); Analysis calculated for C₂₄Hi8BrF4N₇O₂2H₂O.HCl: C, 43.36; H, 3.49; Cl, 5.33; N, 14.75; Found: C, 43.40; H, 3.46; Cl, 5.47; N, 14.64.

Scheme 370

359b 370a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(furan-2-yl)-8-methyl-9H-pyrimido[4,5-b]indol9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (370a) Compound 370a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(fiiran-2-yl)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (359b) (100 mg, 0.182 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (69.5 mg, 0.218 mmol), HATU (138 mg, 0.363 mmol), DIPEA (0.158 mL, 0.908 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4

632 amino-6-(furan-2-yI)-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (370a) (9 mg, 8% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-cfo) δ 10.83 (s, 1H), 8.61 (s, 1H), 8.57 (s, 1H), 8.52 (s, 2H, D₂O exchangeable), 8.01 (d, J= 8.1 Hz, 1H), 7.77 (d, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.64 (s,

1H), 7.31 (d, 7=7.7 Hz, 1 H), 7.03 (d, J = 3.4 Hz, 1H), 6.65 - 6.62 (m, 1H), 5.91 (d, 7=18.0

Hz, 1H), 5.64 (d, J= 17.9 Hz, 1H), 4.43 (dd, J= 9.1, 5.8 Hz, 1H), 3.99 - 3.84 (m, 1H), 2.74 (s, 3H), 2.43-2.12 (m, 2H), 2.01-1.81 (m, 1H), 1.18 - 0.98 (m, 1H), 0.75 - 0.63 (m, 1H); MS (ES+): 586.1 (M+l); (ES-): 584.0 (M-l).

Scheme 371

B(OH)₂

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(pyridin-3-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (371c)

Step-1: Préparation of terA-butyl 2-(4-amino-6-(pyridin-3-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,315 d]pyrimidin-9-yl)acetate (371a)

Compound 371a was prepared according to the procedure reported in in scheme-263 and step-1 of scheme-354, from Zeri-butyl 2-(4-amino-6-bromo-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetate (290e) (300 mg, 0.793 mmol) and pyridin-3-ylboronic acid (146 mg, 1.190 mmol) to afford teri-butyl 2-(4-amino-6-(pyridin-3-yl)-9H-pyrido[2',3^,:4,5]pyrrolo[2,320 d]pyrimidin-9-yl)acetate (371a) (260 mg, 87% yield); Ή NMR (300 MHz, DMSO-d₆) δ 9.08 (d, J= 1.9 Hz, 1H), 8.91 (d, J =1.9 Hz, 1H), 8.65 (dd,J = 4.8, 1.6 Hz, 1H), 8.54 (d, J =1.9

633

Hz, 1H), 8.41 (s, 1H), 8.26 (dt, J = 8.1, 1.9 Hz, 1H), 8.03 (bs, 1H), 7.70 - 7.46 (m, 1H), 6.86 (bs, 1H), 5.23 (s, 2H), 1.41 (s, 9H); MS (ES+): 377 (M+l).

Step-2: Préparation of 2-(4-amino-6-(pyridin-3-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yl)acetic acid (371b)

Compound 371b was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6-(pyridin-3-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetate (371a) (260 mg, 0.691 mmol) in DCM (5 mL) using TFA (788 mg, 6.91 mmol) to afford TFA sait of 2-(4-amino-6-(pyridin-3-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin9-yI)acetic acid (371b) (344 mg) as a pale-yellow solid; *H NMR (300 MHz, DMSO-dé) δ 9.18 (s, 1H), 9.01 (d, J = 3.3 Hz, 1H), 8.71 (d, J = 11.4 Hz, 2H), 8.58 - 8.26 (m, 3H), 7.74 (s, 1H), 7.52 (s, 1H), 5.29 (s, 2H); MS (ES+): 321 (M+l), (ES-): 319 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(pyridin-3-yl)-9Hpyrido[2^,,3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (371c)

Compound 371c was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(pyridin-3-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (371b) (75 mg, 0T73 mmol) in DMF (8 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (66 mg, 0.207 mmol), HATU (131 mg, 0.345 mmol), DIPEA (0.150 mL, 0.863 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4amino-6-(pyridin-3-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)acetyl)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (371c) (47 mg, 47% yield) HCl sait as a pale-yellow solid; *H NMR (300 MHz, DMSO-J₆) δ 10.78 (s, 1H, D₂O exchangeable), 9.45 (d, J= 2.0 Hz, 1H), 9.22 (s, 2H, D₂O exchangeable), 9.15 (d, J= 1.9 Hz, 1H), 9.00 - 8.93 (m, 2H), 8.83 (d, J= 1.9 Hz, 1H), 8.77 (s, 1H), 8.16 (dd, J= 7.9, 5.8 Hz, 1H), 7.95 (d, J= 8.2 Hz, 1H), 7.67 (t, J= 8.0 Hz, 1H), 7.29 (d, J= 7.7 Hz, 1H), 5.91 (d, J = 17.3 Hz, 1H), 5.55 (d, J= 17.2 Hz, 1H), 4.45 (dd, J= 9.0, 5.5 Hz, 1H), 3.97 - 3.89 (m, 1H), 2.42 - 2.15 (m, 2H), 2.00 - 1.85 (m, 1H), 1.17 - 1.03 (m, 1H), 0.88 - 0.76 (m, 1H); MS (ES+): 584.10 (M+l); (ES-): 582.00 (M-l).

Scheme 372

634

371b ^372a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(pyridin-3-yl)-9H-pyrido[2',3':4,5]pynOlo[2,3d]pyrimidin-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (372a)

Compound 372a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(pyridin-3-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (371b) (75 mg, 0.234 mmol) in DMF (8 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (93 mg, 0.281 mmol), HATU (178 mg, 0.468 mmol), DIPEA (0.204 mL, 1.171 mmol) and stirring at

RT for 18 h. This gave after workup and purification as described in scheme-299, ( 1 R,3 S,5R)-2-(2-(4-amino-6-(pyridin-3 -y l)-9H-pyrido [2',3 ' :4,5]pyrrolo [2,3 -d]pyrimidin-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (372a) (48 mg, 34% yield) HCl sait as a pale-yellow solid; Ή NMR (300 MHz, DMSO-ùfe) δ 10.78 (s, 1H, D₂O exchangeable), 9.33 (s, 1H), 9.10 (d, J= 2.8 Hz, 1H), 8.89 (s, 1H), 8.81 15 8.63 (m, 3H), 8.09 - 7.89 (m, 2H), 7.67 (t, J= 8.0 Hz, 1H), 7.29 (d, J= 7.7 Hz, 1H), 5.83 (d,

J= 17.4 Hz, 1H), 5.48 (d, J= 17.3 Hz, 1H), 4.40 (dd, J= 9.0, 5.9 Hz, 1H), 3.73 - 3.63 (m, 1H), 2.50 - 2.41 (m, 1H), 1.99 (dd, J= 13.3, 5.8 Hz, 1H), 1.09 - 1.00 (m, 1H), 1.00 - 0.92 (m, 1H); MS (ES+): 598.1 (M+l); (ES-): 596.1 (M-l).

Scheme 373

635

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-(pyridin-4-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3d]pyrimidin-9-yI)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (373a)

Compound 373a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-(pyridin-4-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetic acid (307b) (80 mg, 0.250 mmol) in DMF (8 mL) using HCl sait of (1R,3S,5R)-N(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (100 mg, 0.300 mmol), HATU (190 mg, 0.500 mmol), DIPEA (0.218 mL, 1.249 mmol) and stirring at

RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6-(pyridm-4-yl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1,0]hexane-3-carboxamide (373a) (72 mg, 48% yield) HCl sait as a pale-yellow solid; ’H NMR (300 MHz, DMSO-î/ô) δ 10.77 (s, 1H, D₂O exchangeable), 9.23 (d, J= 1.9 Hz, 1H), 9.09 - 9.02 (m, 2H), 8.96 (s, 2H,

D₂O exchangeable), 8.84 (d, J= 2.0 Hz, 1H), 8.68 (s, 1H), 8.56 - 8.48 (m, 2H), 8.26 (s, 1H, D₂O exchangeable), 7.97 (d, J= 8.1 Hz, 1H), 7.67 (t, J= 8.0 Hz, 1H), 7.29 (d, J= 7.7 Hz, 1H), 5.84 (d, J= 17.3 Hz, 1H), 5.50 (d, J= 17.2 Hz, 1H), 4.40 (dd, J= 9.1, 5.9 Hz, 1H), 3.69 (dd, J= 5.6, 2.4 Hz, 1H), 2.55-2.40 (m, 1H), 1.99 (dd, J= 13.2, 5.8 Hz, 1H), 1.32 (s, 3H), 1.08 - 1.01 (m, 1H), 1.00 - 0.94 (m, 1H); MS (ES+): 598.10 (M+l); (ES-): 596.0 (M-l).

Scheme 374

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (374g) Step-1: Préparation of N-(2,4-dibromo-5-methoxyphenyl)-2,2,2-trifluoroacetamide (374b)

Compound 374b was prepared according to the procedure reported in step-1 of scheme-46, from 2,4-dibromo-5-methoxyaniline (374a) (5.0 g, 17.80 mmol; CAS # 35736-52-8) in DCM (50 mL) using trifluoroacetic acid anhydride (3.78 mL, 26.68 mmol) and stirring at RT for 1 h. This gave after workup N-(2,4-dibromo-5-methoxyphenyl)-2,2,2-trifluoroacetamide (374b) (6.5 g, 97% yield) as a pale yellow solid; *H NMR (300 MHz, DMSO-îZs) δ 11.37 (s,

1H), 7.99 (s, 1H), 7.27 (s, 1H), 3.86 (s, 3H).

Step-2: Préparation of 2-amino-5-bromo-6-methoxy-lH-indole-3-carbonitrile (374c) Compound 374c was prepared according to the procedure reported in step-1 of scheme-11, from N-(2,4-dibromo-5-methoxyphenyl)-2,2,2-trifluoroacetamide (374b) (6.1 g, 16.18 mmol) in DMSO (24.4 mL) using malononitrile (1.28 g, 19.41 mmol), L-proline (0.37 g, 3.23 mmol), Cul (0.31 g , 1.61 mmol), a solution of K2CO3 (2.80 g, 20.22 mmol) in water (3.05 mL) and heating at 65 °C for 16 h under an argon atmosphère. This gave after workup and

637 purification by flash column chromatography [S1O2 gel, eluting with EtOAc m n-heptane from 0 - 100%] 2-amino-5-bromo-6-methoxy-lH-indole-3-carbonitrile (374c) (1.48 g, 34% yield) as a reddish solid; Ή NMR (300 MHz, DMSO-J₆) δ 10.69 (s, 1H), 7.24 - 7.19 (m, 1H), 6.95 - 6.89 (m, 1H), 6.79 (s, 2H), 3.79 (s, 3H).

Step-3: Préparation of 6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-4-amine (374d) Compound 374d was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-5-bromo-6-methoxy-lH-indole-3-carbonitrile (374c) (1.6 g, 6.01 mmol) using trimethyl orthoformate (19.73 mL, 180.3 mmol), AcOH (1.72 mL, 30.05 mmol) and NH4OAC (2.32 g, 30.05) at RT. This gave after workup AcOH sait of 6-bromo-7-methoxy9H-pyrimido[4,5-b]indol-4-amine (374d) (1.65 g, 78% yield) as a greenish solid; MS (ES-) 291.3 (M-l).

Step-4: Préparation of ZerZ-butyl 2-(4-amino-6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetate (374e)

Compound 374e was prepared according to the procedure reported in step-2 of scheme-16, from AcOH sait of 6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-4-amine (374d) (1.65 g, 4.67 mmol) in DMF (66 mL) using ZerZ-butyl 2-bromoacetate (0.83 mL, 5.62 mmol) and CS2CO3 (3.67 g, 11.24 mmol) and stirring at RT for 16 h. This gave after workup and purification by flash column chromatography [S1O2 gel, eluting with MeOH in DCM from 0 10%] ZerZ-butyl 2-(4-amino-6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (374e) (0.97 g, 51% yield ) as an off white solid; ’H NMR (300 MHz, DMSO-cfe) δ 8.62 (s, 1H), 8.24 (s, 1H), 7.40 (s, 1H), 7.27 (s, 2H), 5.14 (s, 2H), 3.92 (s, 3H), 1.40 (s, 9H).

Step-5: Préparation of 2-(4-amino-6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (374f)

Compound 374f was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (374e) (0.97 g, 2.38 mmol) in DCM (4.0 mL) using TFA (4.0 mL, 52.36 mmol) in DCM (11.60 mL) to afford 2-(4-amino-6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (374f) (1.1 g, 99% yield) TFA sait as an off white solid; ’H NMR (300 MHz, DMSO-î/ê) δ 8.80 (s, 1H), 8.52 (s, 1H), 8.44 (s, 2H), 7.62 (s, 1H), 5.27 (s, 2H), 3.96 (s, 3H).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-7-methoxy-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (374g)

638

Compound 374g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (374f) (87 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (57.3 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (0.131 mL, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6bromo-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (374g) (67 mg, 73% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 10.71 (s, 1H, D₂O exchangeable), 8.82 (s, 1H), 8.56 (s, 1H), 8.53 (s, 3H, D₂O exchangeable), 8.00 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.42 (s, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.72 (d, J= 17.3 Hz, 1H), 5.46 (d, J= 17.2 Hz, 1H), 4.41 (dd, J= 9.1, 5.7 Hz, 1H), 3.97 (s, 3H), 3.94 - 3.86 (m, 1H), 2.42 - 2.15 (m, 2H), 2.01 - 1.79 (m, 1H), 1.13 - 0.97 (m, 1H), 0.82 - 0.67 (m, 1H); MS (ES+): 614/616 (M+l).

Scheme 375

H N. .Br

HATU, DIPEA

Br 375a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (375a)

Compound 375a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (374f) (87 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (59.9 mg, 0.1800 mmol), HATU (114 mg, 0.300 mmol), DIPEA (0.131 mL, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4amino-6-bromo-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (375a) (47 mg, 50% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-Jô) δ 10.70 (s, 1H, D₂O exchangeable), 8.81 (s, 1H),

639

8.55 (s, 1H), 8.51 (s, 2H, D₂0 exchangeable), 8.01 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.42 (s, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.68 (d, J= 17.4 Hz, 1H), 5.41 (d, J= 17.2 Hz, 1H), 4.36 (dd, J =9.1,6.0 Hz, 1H), 3.96 (s, 3H), 3.73 - 3.63 (m, 1H), 2.56-2.40 (m, 1H), 1.99 (dd, J= 13.1, 6.1 Hz, 1H), 1.32 (s, 3H), 1.07 - 0.96 (m, 1H), 0.94 - 0.87 (m, 1H); MS

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-8-fluoro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (376g)

Step-1 : Préparation of N-(2,4-dibromo-6-fluorophenyl)-2,2,2-trifluoroacetamide (376b)

Compound 376b was prepared according to the procedure reported in step-1 of scheme-46, from 2,4-dibromo-6-fluoroaniline (376a) (10.0 g, 37.19 mmol; CAS # 141474-37-5) in DCM (100 mL) using trifluoroacetic acid anhydride (8.4 mL, 55.78 mmol) to afford N-(2,4dibromo-6-fluorophenyl)-2,2,2-trifluoroacetamide (376b) (13.27 g, 98% yield) as an off white solid; *H NMR (300 MHz, DMSO-7₆) δ 11.55 (s, 1H), 7.97 (t, J= 1.9 Hz, 1H), 7.87 (dd, 7= 9.1, 2.1 Hz, 1H).

640

Step-2: Préparation of 2-amino-5-bromo-7-fluoro-lH-indole-3-carbonitrile (376c)

Compound 376c was prepared according to the procedure reported in step-1 of scheme-11 and step-2 of scheme-374, from N-(2,4-dibromo-6-fluorophenyl)-2,2,2-trifluoroacetamide (376b) (7.40 g, 20.28 mmol) in DMSO (20.0 mL) using malononitrile (1.35 mL, 24.32 mmol), L-proline (0.47 mg, 4.06 mmol), Cul (386 mg, 2.03 mmol), a solution of K2CO3 (3.50 g, 25.34 mmol) in water (1.4 mL) to afford 2-amino-5-bromo-7-fluoro-lH-indole-3carbonitrile (376c) (1.84 g, 36% yield) as a brown solid; 'H NMR (300 MHz, DMSO-J₆) δ 11.57 (s, 1H), 7.14-7.07 (m, 1H), 7.06-7.00 (m, 1H), 6.97 (s, 2H).

Step-3: Préparation of 6-bromo-8-fluoro-9H-pyrimido[4,5-b]indol-4-amine (376d)

Compound 376d was prepared according to the procedure reported in step-1 of scheme-6 and step-3 of scheme-374, from 2-amino-5-bromo-7-fluoro-lH-indole-3-carbonitrile (376c) (3.40 g, 13.38 mmol) using trimethyl orthoformate (42.61 g, 401.47 mmol), AcOH (4.017 g, 66.9 mmol), NH4OAC (5.156 g, 66.9 mmol) to afford AcOH sait of 6-bromo-8-fluoro-9Hpyrimido[4,5-b]indol-4-amine (376d) (3.50 g, 93% yield) as an off white solid; *H NMR (300 MHz, DMSO-î/ô) δ 12.54 (s, 1H), 8.46 (d, J= 1.6 Hz, 1H), 8.30 (s, 1H), 7.50 (dd, J= 10.4, 1.5 Hz, 1H), 7.42 (s, 2H).

Step-4: Préparation of tert-butyl 2-(4-amino-6-bromo-8-fluoro-9H-pyrimido[4,5-b]indol-9yl)acetate (376e)

Compound 376e was prepared according to the procedure reported in step-2 of scheme-16 and step-4 of scheme-374, from AcOH sait of 6-bromo-8-fluoro-9H-pyrimido[4,5-b]indol-4amine (376d) (3.50 g, 12.45 mmol) in DMF (140 mL) using tert-butyl 2-bromoacetate (2.43 g, 12.45 mmol), CS2CO3 (8.11 g, 24.90 mmol) to afford tert-butyl 2-(4-amino-6-bromo-8fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (376e) (3.10 g, 63 % yield) as an off white solid; Ή NMR (300 MHz, DMSO-J₆) δ 8.51 (d, J= 1.6 Hz, 1H), 8.35 (s, 1H), 7.68 - 7.46 (m, 3H), 5.25 - 5.07 (m, 2H), 1.40 (s, 9H).

Step-5: Préparation of 2-(4-amino-6-bromo-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (376f)

Compound 376f was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-6-bromo-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (376e) (3.0 g, 7.59 mmol) in DCM (50 mL) using TFA (12.77 mL, 166.99 mmol) to afford 2-(4amino-6-bromo-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (376f) (3.5 g) TFA sait as

641 an off white solid; Ή NMR (300 MHz, DMSO-î/ô) δ 8.60 (d, J= 1.7 Hz, 1H), 8.49 (s, 1H), 8.16 (s, 2H), 7.64 (dd, J= 11.7, 1.6 Hz, 1H), 5.22 (d, J= 1.9 Hz, 2H).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-8-fluoro-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (376g)

Compound 376g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (376f) (85 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (57.3 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (0.131 mL, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6bromo-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (376g) (52 mg, 58% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î/ô) δ 10.76 (s, 1H), 8.61 (s, 1H), 8.55 (s, 1H), 8.32 (s, 2H, D₂O exchangeable), 8.08 - 7.92 (m, 1H), 7.79 - 7.56 (m, 2H), 7.39 - 7.22 (m, 1H), 5.76 (d, J = 17.7 Hz, 1H), 5.42 (d, J= 17.4 Hz, 1H), 4.53 - 4.26 (m, 1H), 3.95 - 3.77 (m, 1H), 2.43 2.13 (m, 2H), 2.02-1.81 (m, 1H), 1.14-1.01 (m, 1H), 0.72-0.50 (m, 1H); MS (ES+): 602.00 & 604.00 (M+l).

Scheme 377

376f ^377a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-8-fluoro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (377a)

Compound 377a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid

(376f) (85 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (59.9 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (0.131 mL, 0.750 mmol) and stirring atRT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(45 amino-6-bromo-8-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (377a) (84 mg, 91% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-t/₆) δ 10.77 (s, 1H), 8.70 - 8.49 (m, 4H, 2H D₂O exchangeable), 8.05 - 7.96 (m, 1H), 7.78 - 7.55 (m, 2H), 7.37 - 7.23 (m, 1H), 5.72 (d, J= 17.5 Hz, 1H), 5.38 (d, J= 17.5 Hz, 1H), 4.43-4.32 (m, 1H), 3.13-2.98 (m, 1H), 2.61 10 2.32 (m, 1H), 2.17-1.76 (m, 1H), 1.29 (s, 3H), 1.08 - 0.95 (m, 1H), 0.83 - 0.70 (m, 1H); ¹⁹F

NMR (282 MHz, DMSO-îZ₆) δ -131.77; MS (ES+): 616.00 & 618.00 (M+l).

Scheme 378

Br

192e 378a

378b 378c

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(pyridin-2-yl)-9H-pyrimido[4,515 b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3carboxamide (378c)

Step-1: Préparation of ZerZ-butyl 2-(4-amino-8-methyl-6-(pyridin-2-yl)-9H-pyrimido[4,5b]indol-9-yl)acetate (378a)

643

A mixture of tert-butyl 2-(4-amino-6-bromo-8-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (192e) (300 mg, 0.767 mmol), B2(pin)2 (292 mg, 1.150 mmol), Pd(dppf)C12.CH2C12 (62.6 mg, 0.077 mmol), KOAc (226 mg, 2.300 mmol) in anhydrous dioxane (15 mL) was heated at 100 °C for 16 h under nitrogen. To this mixture 2-bromopyridine (182 mg, 1.150 mmol), Pd(PPh3)2Ch (53.8 mg, 0.077 mmol) and 3.3 M aqueous K2CO3 (0.697 mL, 2.300 mmol) were added and the mixture was stirred at 100 °C for 16 h under nitrogen. The resulting mixture was diluted with EtOAc (50 mL) and fîltered. The filtrate was washed with H2O (25 mL x 3), brine (25 mL), dried, fîltered and concentrated in vacuum. The residue obtained was purified using flash column chromatography [S1O2 gel (12 g), eluting with DMA-80 in DCM from 0-20%] to provide teri-butyl 2-(4-amino-8-methyl-6-(pyridin-2-yl)-9H-pyrimido[4,5b]indol-9-yl)acetate (378a) (60 mg, 20% yield) as a yellow semi-solid; ’H NMR (300 MHz, DMSO-àfe) δ 8.75 (d, J= 3.7 Hz, 1H), 8.66 (s, 1H), 8.43 - 8.21 (m, 2H), 8.08 (s, 1H), 7.92 (t, J= 8.7 Hz, 1H), 7.48 (s, 2H), 7.32 (d, J= 6.6 Hz, 1H), 5.37 (s, 2H), 2.71 (s, 3H), 1.44 (s, 9H); MS (ES+): 390 (M+l), (ES-): 388 (M-l).

Step-2: Préparation of 2-(4-amino-8-methyl-6-(pyridin-2-yl)-9H-pyrimido[4,5-b]indol-9yl)acetic acid (378b)

Compound 378b was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-8-methyI-6-(pyridin-2-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetate (378a) (60 mg, 0.154 mmol) in DCM (5 mL) using TFA (176 mg, 1.541 mmol) to afford after purification as reported in step-2 of scheme-319, 2-(4-amino-8-methyl-6-(pyridin-2-yl)9H-pyrimido[4,5-b]indol-9-yl)acetic acid (378b) (33 mg, 58% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 9.23 (s, 1H), 9.15 (bs, 2H), 8.81 (dd, J= 5.5, 1.7 Hz, 1H), 8.71 (s, 1H), 8.57 (d, J = 8.2 Hz, 1H), 8.38 (t, J= 7.8 Hz, 1H), 8.21 (s, 1H), 7.84-7.66 (m, 1H), 5.52 (s, 2H), 2.79 (s, 3H); MS (ES+): 334 (M+l), (ES-): 332 (M-l).

Step-3: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-methyl-6-(pyridin-2-yl)-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (378c)

Compound 378c was prepared according to the procedure reported in step-3 of scheme-1, from HCl sait of 2-(4-amino-8-methyl-6-(pyridin-2-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (378b) (30 mg, 0.081 mmol) in DMF (5 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (32.4 mg, 0.097 mmol), HATU (61.7 mg, 0.162 mmol), DIPEA (0.071 mL, 0.406 mmol) and stirring at RT

644 for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2(2-(4-amino-8-methyl-6-(pyridin-2-yl)-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicycIo[3.1.0]hexane-3-carboxamide (378c) (32 mg, 65% yield) HCl sait as a white solid; Y NMR (300 MHz, DMSO-J₆) δ 10.84 (s, 1H), 9.08 (s, 1H), 5 8.95 (s, 2H, D₂O exchangeable), 8.80 - 8.74 (m, 1H), 8.68 (s, 1H), 8.43 (d, J= 8.1 Hz, 1H),

8.24 (t, J= 7.8 Hz, 1H), 8.14 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.66 7.58 (m, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.94 (d, J= 18.0 Hz, 1H), 5.65 (d, J= 17.9 Hz, 1H), 4.40 (dd, J= 9.0, 6.2 Hz, 1H), 3.72 (dd, J= 5.5, 2.4 Hz, 1H), 2.79 (s, 3H), 2.56 - 2.42 (m, 1H), 1.99 (dd, J= 13.2, 6.0 Hz, 1H), 1.31 (s, 3H), 1.08-0.99 (m, 1H), 0.89-0.83 (m, 1H);

MS (ES+): 611.10 (M+l); (ES-): 609.10 (M-l).

Scheme 379

379g

379h

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-7-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (379h)

Step-1: Préparation ofN-(4-bromo-3-methylphenyl)-2,2,2-trifluoroacetamide (379b)

645

Compound 379b was prepared according to the procedure reported in step-1 of scheme-46, from 4-bromo-3-methylaniline (379a) ( 15.0 g, 80.62 mmol; CAS # 6933-10-4) in DCM (30 mL) using trifluoroacetic acid anhydride (25.40 g, 120.93 mmol) and stirring at RT for 30 min. This gave after workup N-(4-bromo-3-methylphenyl)-2,2,2-trifluoroacetamide (379b) (22.8 g) as a brown solid; Ή NMR (300 MHz, DMSO-cfe) δ 11.31 (s, 1H), 7.88 - 7.54 (m, 2H), 7.45 (dd, J= 8.6, 2.6 Hz, 1H), 2.35 (s, 3H).

Step-2: Préparation of N-(4-bromo-2-iodo-5-methylphenyl)-2,2,2-trifluoroacetamide (379c)

To a stirred solution of N-(4-bromo-3-methylphenyl)-2,2,2-trifluoroacetamide (379b) (22.8 g, 80.83 mmol) in acetonitrile (228 mL) was added at RT NIS (18.18 g, 80.83 mmol), triflic acid (0.6 g, 4.05 mmol) and stirred ovemight at RT. Reaction quenched with water (250.0 mL), extracted with ethyl acetate (2 X 250 mL). The combined organic layers were washed with brine (150 mL), dried, filtered and concentrated to give N-(4-bromo-2-iodo-5methylphenyl)-2,2,2-trifluoroacetamide (379c) (31.4 g, 95% yield) as a brown solid; 'H NMR (300 MHz, DMSO-<7₆) δ 11.29 (s, 1H), 8.13 (s, 1H), 7.41 (s, 1H), 2.31 (s, 3H).

Step-3: Préparation of 2-amino-5-bromo-6-methyl-lH-indole-3-carbonitrile (379d)

Compound 379d was prepared according to the procedure reported in step-1 of scheme-11 and step-2 of scheme-374, from N-(4-bromo-2-iodo-5-methylphenyl)-2,2,2trifluoroacetamide (379c) (10.0 g, 24.51 mmol) in DMSO (40.0 mL) using malononitrile (1.94 g, 29.41 mmol), L-proline (0.569 g, 4.92 mmol), Cul (0.47 g , 2.45 mmol), a solution of K2CO3 (4.064 g, 29.41 mmol) in water (0.5 mL) to afford 2-amino-5-bromo-6-methyl-lHindole-3-carbonitrile (379d) (4.0 g, 65% yield) as a yellow solid; ’H NMR (300 MHz, DMSO-îZô) δ 10.77 (s, 1H), 7.25-7.23 (m, 1H), 7.09 (s, 1H), 6.88 (s, 2H), 2.35-2.31 (m, 3H).

Step-4: Préparation of 6-bromo-7-methyl-9H-pyrimido[4,5-b]indol-4-amine (379e)

Compound 379e was prepared according to the procedure reported in step-1 of scheme-6, from 2-amino-5-bromo-6-methyl-lH-indole-3-carbonitrile (379d)( 4.0 g, 15.99 mmol) using trimethyl orthoformate (50.90 g, 479.7 mmol), AcOH (4.80 g) and NH₄OAc (6.16 g, 79.96 mmol) at 100 °C for 16 h. This gave after workup AcOH sait of 6-bromo-7-methyl-9Hpyrimido[4,5-b]indol-4-amine (379e) (3.13 g, 58% yield) as a gray solid; MS (ES+): 277.1 (M+l).

646

Step-5: Préparation of ZerZ-butyl 2-(4-amino-6-bromo-7-methyl-9H-pynmido[4,5-b]indol-9yl)acetate (379f)

Compound 379f was prepared according to the procedure reported in step-2 of scheme-16 and step-4 of scheme-374, from AcOH sait of 6-bromo-7-methyl-9H-pyrimido[4,5-b]indol-4amine (379e) (3.13 g, 9.28 mmol) in DMF (125.2 mL) using ZerZ-butyl 2-bromoacetate (2.2 g, 11.29 mmol), CS2CO3 (7.35 g, 22.59 mmol) to afford ZerZ-butyl 2-(4-amino-6-bromo-7methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (379f) (1.32 g, 36% yield) as a yellow solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.64 (s, 1H), 8.27 (d, J= 0.8 Hz, 1H), 7.60 (s, 1H), 7.36 (s, 2H), 5.07 (s, 2H), 2.49 (s, 3H), 1.57 - 1.30 (m, 9H).

Step-6: Préparation of 2-(4-amino-6-bromo-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (379g)

Compound 379g was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6-bromo-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (379f) (1.3 g, 3.32 mmol) in DCM (10 mL) using TFA (8.33 g, 73.10 mmol) in DCM (15 mL) to afford 2-(4-amino-6-bromo-7-methyl-9H-pyrimido[4,5-b]indoI-9-yl)acetic acid (379g) (1.35 g, 91% yield) TFA sait as an off white solid; 'H NMR (300 MHz, DMSO-î/₆) δ 8.60 (s, 1H), 8.45 (s, 1H), 7.62 (s, 1H), 5.16 (s, 2H), 2.45 (s, 3H).

Step-7: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-7-methyl-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (379h)

Compound 379h was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (379g) (84 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (57.3 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (0.131 mL, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4amino-6-bromo-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (379h) (45 mg, 50% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-î7₆) δ 10.76 (s, 1H), 8.80 (s, 1H), 8.56 (s, 1H), 8.48 (s, 3H, D₂O exchangeable), 8.06 - 7.93 (m, 1H), 7.76 - 7.62 (m, 2H), 7.36 - 7.25 (m, 1H), 5.70 (d, J= 17.6 Hz, 1H), 5.37 (d, J= 17.9 Hz, 1H), 4.52-4.31 (m, 1H), 3.99 - 3.82 (m, 1H), 2.4121436

647

2.13 (m, 2H), 2.00- 1.81 (m, 1H), 1.11 -0.97 (m, 1H), 0.88 - 0.62 (m, 1H); MS (ES+):

598.00 (M+l); (ES-): 595.95 (M-l).

Scheme 380

H N. .Br

HATU, DIPEA

379g

380a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-7-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (380a)

Compound 380a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (379g) (84 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (59.9 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (0.131 mL, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2(4-amino-6-bromo-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (380a) (73 mg, 79% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-cfc) δ 10.76 (s, 1H, D2O exchangeable), 8.80 (s, 1H), 8.70 - 8.47 (m, 3H, 2H D2O exchangeable), 8.06 - 7.94 (m, 1H), 7.77 - 7.58 (m, 2H), 7.35-7.25 (m, 1H), 5.65 (d, J= 17.2, 3.8 Hz, 1H), 5.31 (d,J=17.4 Hz, 1H), 4.42-4.30 (m, 1H), 3.70-3.61 (m, 1H), 2.65-2.33 (m, 4H), 2.09-1.89 (m, 1H), 1.30 (s, 3H), 1.07-0.87 (m, 2H); MS (ES+): 612.00 (M+l); (ES-): 609.90 (M-l).

Scheme 381

648

381d ^381e

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-fluoro-7-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (381g)

Step-1: Préparation ofN-(2-bromo-4-fluoro-5-methylphenyl)-2,2,2-trifluoroacetamide (381b)

Compound 381b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-4-fluoro-5-methylaniline (381a)( 4.0 g, 19.60 mmol; CAS # 1065076-39-2) in DCM (20 mL) using trifluoroacetic acid anhydride (6.17g, 29.40 mmol) and stirring at RT for 1 h. This gave after workup N-(2-bromo-4-fluoro-5-methylphenyl)-2,2,2-trifluoroacetamide (381b) (6.25 g) as a light violet color solid; Ή NMR (300 MHz, DMSO-î7₆) δ 11.26 (s, 1H),

7.66 (d, J= 9.1 Hz, 1H), 7.43 (d, J= Ί.6 Hz, 1H), 2.22 (d, J= 2.1 Hz, 3H).

Step-2: Préparation of 2-amino-5-fluoro-6-methyl-lH-indole-3-carbonitrile (381c)

Compound 381c was prepared according to the procedure reported in step-1 of scheme-11 and step-2 of scheme-374, from N-(2-bromo-4-fluoro-5-methylphenyl)-2,2,2trifluoroacetamide (381b) (6.25 g, 20.83 mmol) in DMSO (16 mL) using malononitrile (1.65

649 g, 24.99 mmol), L-proline (0.48 g, 4.16 mmol), Cul (0.40 g , 2.08 mmol), a solution of K2CO3 (5.75 g, 41.64 mmol) in water (16 mL) to afford 2-amino-5-fluoro-6-methyl-lHindole-3-carbonitrile (381c) (1.4 g, 36% yield) as a gray solid; ’HNMR (300 MHz, DMSOd₆) δ 10.64 (s, 1H), 6.97 (d, J= 6.5 Hz, 1H), 6.82 (d, J= 10.2 Hz, 1H), 6.77 (s, 2H), 2.21 (d, J = 2.2 Hz, 3H).

Step-3: Préparation of 6-fluoro-7-methyl-9H-pyrimido[4,5-b]indol-4-amine (381d)

Compound 381d was prepared according to the procedure reported in step-1 of scheme-6 and step-3 of scheme-374, from 2-amino-5-fluoro-6-methyl-lH-indole-3-carbonitrile (381c) (1.35 g, 7.14 mmol) using trimethyl orthoformate (22.69 g, 213.9 mmol), AcOH (2.03 mL) NH4OAC (2.75 g, 35.67 mmol) to afford AcOH sait of 6-fluoro-7-methyl-9H-pyrimido[4,5b]indol-4-amine (381d) (1.0 g, 65% yield) as a gray solid; 'H NMR (300 MHz, DMSO-dô) δ 11.77 (s, 1H), 8.22 (s, 1H), 8.15 (d, J= 10.9 Hz, 1H), 7.29 (d, J= 6.6 Hz, 1H), 7.16 (s, 2H), 2.37 (d, J =2.2 Hz, 3H).

Step-4: Préparation of ZerZ-butyl 2-(4-amino-6-fluoro-7-methyl-9H-pyrimido[4,5-b]indol-9yl)acetate (381e)

Compound 381e was prepared according to the procedure reported in step-2 of scheme-16 and step-2 of scheme-374, from AcOH sait of 6-fluoro-7-methyl-9H-pyrimido[4,5-b]indol-4amine (381d) (1.0 g, 3.82 mmol) in DMF (40 mL) using ZerZ-butyl 2-bromoacetate (0.745 g, 3.82 mmol), CS2CO3 (2.49 g, 7.65 mmol) to afford ZerZ-butyl 2-(4-amino-6-fluoro-7-methyl9H-pyrimido[4,5-b]indol-9-yl)acetate (381e) (1.21 g, 80% yield) as an off white solid; *H NMR (300 MHz, DMSO-ri₆) δ 8.26 (s, 1H), 8.21 (d, J= 10.7 Hz, 1H), 7.47 (d, J= 6.4 Hz, 1H), 7.29 (s, 2H), 5.07 (s, 2H), 2.39 (d, J= 22 Hz, 3H), 1.40 (s, 9H).

Step-5: Préparation of 2-(4-amino-6-fluoro-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (381f)

Compound 381f was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6-fluoro-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (381e) (1.44 g, 4.36 mmol) in DCM (6 mL) using TFA (10.93 g, 95.90 mmol) in DCM (18 mL) to afford 2-(4-amino-6-fluoro-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (381f) (1.62 g, 96% yield) TFA sait as an off white solid; Ή NMR (300 MHz, DMSO-J₆) δ 8.71 (s, 2H), 8.60 (s, 1H), 8.37 (d, J= 10.5 Hz, 1H), 7.74 (d, J= 6.3 Hz, 1H), 5.23 (s, 2H), 2.42 (d, J= 2.2 Hz, 3H).

650

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-fluoro-7-methyl-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1,0]hexane-3-carboxamide (381g)

Compound 381g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-fluoro-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (381f) (75 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (57.3 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (0.131 mL, 0.750 mmol) and stirring at RT for 22 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6fluoro-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicycIo[3.1.0]hexane-3-carboxamide (381g) (36 mg, 45% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/₆) δ 10.77 (s, 1H), 8.66 (s, 2H), 8.61 (s, 1H), 8.41 (d, J= 10.4 Hz, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.64 (d, J= 6.3 Hz, 1H), 7.32 (d, J = 7.8 Hz, 1H), 5.73 (d, J= 17.3 Hz, 1H), 5.39 (d, J= 17.3 Hz, 1H), 4.42 (dd, J= 9.1, 5.6 Hz, 1H), 3.97-3.83 (m, 1H), 2.42 (s, 3H), 2.39-2.16 (m, 2H), 1.99-1.85 (m, 1H), 1.13-1.00 (m, 1H), 0.84-0.75 (m, 1H); MS (ES+): 538.1 (M+l); (ES-): 536.0 (M-l).

Scheme 382

H

N. ^N. ^Br

HATU, DIPEA

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-fluoro-7-methyI-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (382a)

Compound 382a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-fluoro-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (3811) (75 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (59.9 mg, 0.180 mmol), HATU (114 mg, 0.3 mmol), DIPEA (0.131 mL, 0.750 mmol) and stirring atRT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(421436

amino-6-fluoro-7-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (382a) (77 mg, 93% yield) HCl sait as a white solid; 'HNMR (300 MHz, DMSO-t7₆) δ 10.77 (s, 1H), 8.67 (s, 2H, D₂O exchangeable), 8.60 (s, 1H), 8.41 (d, J= 10.4 Hz, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.64 (d, J= 6.4 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.69 (d, J= 17.4 Hz, 1H), 5.34 (d, J= 17.3 Hz,

1H), 4.42 - 4.31 (m, 1H), 3.67 (d, J= 6.1 Hz, 1H), 2.47 - 2.43 (m, 1H), 2.42 (s, 3H), 2.04 1.91 (m, 1H), 1.31 (s, 3H), 1.06 - 0.87 (m, 2H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -123.40; MS (ES+): 552.10 (M+l); (ES-): 550.0 (M-l).

Scheme 383

383e

383f 383g

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (383g)

Step-1 : Préparation of N-(2-bromo-4-fluoro-5-methoxyphenyl)-2,2,2-trifluoroacetamide (383b)

Compound 383b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-4-fluoro-5-methoxyaniline (383a)( 5.0 g, 22.72 mmol; CAS # 420786-92-1) in

652

DCM (50 mL) using trifluoroacetic acid anhydride (4.80 mL, 34.08 mmol) and stirring at RT for 1 h. This gave after workup N-(2-bromo-4-fluoro-5-methoxyphenyl)-2,2,2trifluoroacetamide (383b) (7.0 g, 97% yield) as a light purple solid; ’H NMR (300 MHz, DMSO-7₆)Ô 11.29 (s, 1H), 7.73 (dd, 7= 10.8, 1.2 Hz, 1H), 7.34 (dd, 7= 8.5, 1.2 Hz, 1H), 3.84 (d, 7= 1.3 Hz, 3H).

Step-2: Préparation of 2-amino-5-fluoro-6-methoxy-lH-indole-3-carbonitrile (383c)

Compound 383c was prepared according to the procedure reported in step-1 of scheme-11 and step-2 of scheme-374, from N-(2-bromo-4-fluoro-5-methoxyphenyl)-2,2,2trifluoroacetamide (383b) (6.5 g, 20.57 mmol) in DMSO (26 mL) using malononitrile (1.63 g, 24.67 mmol), L-proline (0.47 g, 4.11 mmol), Cul (0.39 g , 2.05 mmol), a solution of K2CO3 (4.26 g, 30.84 mmol) in water (6.5 mL) to afford 2-amino-5-fluoro-6-methoxy-lHindole-3-carbonitrile (383c) (1.42 g, 34% yield) as a black solid; 'H NMR (300 MHz, DMSO-7₆) δ 10.58 (s, 1H), 6.94 (s, 1H), 6.91 (d, 7= 4.3 Hz, 1H), 6.68 (s, 2H), 3.78 (s, 3H).

Step-3: Préparation of 6-fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-4-amine (383d)

Compound 383d was prepared according to the procedure reported in step-1 of scheme-6 and step-3 of scheme-374, from 2-amino-5-fluoro-6-methoxy-lH-indole-3-carbonitrile (383c) (1.48 g, 7.21 mmol) using trimethyl orthoformate (23.67 mL, 216.3 mmol), AcOH (2.06 mL, 36.05 mmol), NH4OAC (2.78 g, 36.05 mmol) to afford 6-fluoro-7-methoxy-9H-pyrimido[4,5b]indol-4-amine (383d) (1.20 g, 72% yield) as a light brown solid; *H NMR (300 MHz, DMSO-Jô) δ 11.80 (s, 1H), 8.40 - 8.10 (m, 2H), 7.22-6.98 (m, 3H), 3.91 (s, 3H).

Step-4: Préparation of terf-butyl 2-(4-amino-6-fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-9yl)acetate (383e)

Compound 383e was prepared according to the procedure reported in step-2 of scheme-16 and step-4 of scheme-374, from 6-fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-4-amine (383d) (1.20 g, 5.17 mmol,) in DMF (48 mL) using terributyl 2-bromoacetate (0.95 mL, 6.19 mmol), CS2CO3 (3.86 g, 11.35 mmol) to afford ZerZ-butyl 2-(4-amino-6-fluoro-7-methoxy9H-pyrimido[4,5-b]indol-9-yl)acetate (383e) (0.95 g, 53% yield) as an off white solid; *H NMR (300 MHz, DMSO-7₆) δ 8.28 (d, 7= 12.1 Hz, 1H), 8.23 (s, 1H), 7.44 (d, J= 7.4 Hz, 1H), 7.22 (s, 2H), 5.13 (s, 2H), 3.92 (s, 3H), 1.40 (s, 10H).

Step-5: Préparation of 2-(4-amino-6-fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (383f)

653

Compound 383f was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-amino-6-fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetate (383e) (0.95 g, 2.74 mmol) in DCM (4.75 mL) using TFA (4.62 mL, 60.28 mmol) in DCM (10.45 mL) to afford 2-(4-amino-6-fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (383f) (1.10 g, 99.2% yield) TFA sait as an off white solid; Ή NMR (300 MHz, DMSO-ûfc) δ 8.62 - 8.51 (m, 3H), 8.47 (d, J= 11.9 Hz, 1H), 7.71 (d, J= 7.3 Hz, 1H), 5.29 (s, 2H), 3.96 (s, 3H). .

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-fluoro-7-methoxy-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (383g)

Compound 383g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (383f) (78 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (57.3 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (0.131 mL, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (383g) (36 mg, 43% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 10.72 (s, 1H), 8.55 (s, 1H), 8.52 - 8.43 (m, 3H, 2H D₂O exchangeable), 8.00 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 7.9 Hz, 1H), 7.49 (d, J= 7.2 Hz, 1H), 7.33 (d, J= 7.7 Hz, 1H), 5.72 (d, J= 17.3 Hz, 1H), 5.46 (d, J= 17.3 Hz, 1H), 4.48-4.37 (m, 1H), 3.97 (s, 3H), 3.93 - 3.82 (m, 1H), 2.40-2.13 (m, 2H), 2.00 - 1.83 (m, 1H), 1.15 - 0.96 (m, 1H), 0.85 - 0.66 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î/₆) δ -140.13; MS (ES+):554.10 (M+l); (ES-):552.0 (M-l).

Scheme 384

654

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (384g)

Step-1: Préparation ofN-(2-bromo-5-fluoro-4-methylphenyl)-2,2,2-trifluoroacetamide (384b)

Compound 384b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-5-fhioro-4-methylaniline (384a) ( 3.0 g, 14.70 mmol; CAS # 202865-78-9) in DCM (30 mL) using trifluoroacetic acid anhydride (4.63 g, 22.05 mmol) and stirring at RT for 1 h. This gave after workup N-(2-bromo-5-fluoro-4-methylphenyl)-2,2,2trifluoroacetamide (384b) (3.5 g, 79% yield) as a dark brown solid; 'H NMR (300 MHz, DMSO-7,) δ 11.33 (s, 1H), 7.73 (d, J= 7.7 Hz, 1H), 7.39 (d, J= 10.0 Hz, 1H), 2.26 (d, J= 2.0 Hz, 3H).

Step-2: Préparation of 2-amino-6-fluoro-5-methyl-lH-indole-3-carbonitrile (384c)

Compound 384c was prepared according to the procedure reported in step-1 of scheme-11 and step-2 of scheme-374, from N-(2-bromo-5-fluoro-4-methylphenyl)-2,2,2trifluoroacetamide (384b) (5.0 g, 16.66 mmol) in DMSO (15 mL) using malononitrile (1.32 g, 19.98 mmol), L-proline (0.38 g, 3.32 mmol), Cul (0.31 g , 1.66 mmol), a solution of K2CO3 (4.60 g, 33.35 mmol) in water (15 mL) to afford 2-amino-6-fluoro-5-methyl-lHindole-3-carbonitrile (384c) (1.5 g, 48% yield) as a white solid; Ή NMR (300 MHz, DMSOd₆) δ 10.62 (s, 1H), 6.96 (d, J= 7.1 Hz, 1H), 6.91 (d, J= 10.3 Hz, 1H), 6.73 (s, 2H), 2.22 (s, 3H).

Step-3: Préparation of 7-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-4-amine (384d)

655

Compound 384d was prepared according to the procedure reported in step-1 of scheme-6 and step-3 of scheme-374, from 2-amino-6-fluoro-5-methyl-lH-indole-3-carbonitrile (384c) (1.5 g, 7.93 mmol) using trimethyl orthoformate (25.24 g, 237.85 mmol), AcOH (7.5 mL), NH4OAC (3.05 g, 39.65 mmol) to afford 7-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-4-amine (384d) ( 1.2 g, 70% yield) as an off-white solid.

Step-4: Préparation of ZerZ-butyl 2-(4-amino-7-fluoro-6-methyI-9H-pyrimido[4,5-b]indol-9yl)acetate (384e)

Compound 384e was prepared according to the procedure reported in step-2 of scheme-16 and step-4 of scheme-374, from 7-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-4-amine (384d) (1.2 g, 5.55 mmol) in DMF (36.0 mL) using ZerZ-butyl 2-bromoacetate (1.07 g, 5.55 mmol), CS2CO3 (3.60 g, 11.09 mmol) to afford ZerZ-butyl 2-(4-amino-7-fluoro-6-methyl-9Hpyrimido[4,5-b]indol-9-yl)acetate (384e) (500.0 mg, 27% yield ) as an off-white solid; 'H NMR (300 MHz, DMSO-d₆) δ 8.30 - 8.24 (m, 2H), 7.47 (d, J= 10.7 Hz, 1H), 7.25 (s, 2H), 5.08 (s, 2H), 2.37 (d, J= 2.2 Hz, 3H), 1.40 (s, 9H).

Step-5: Préparation of 2-(4-amino-7-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (384f)

Compound 384f was prepared according to the procedure reported in step-2 of scheme-1, from tezY-butyl 2-(4-amino-7-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (384e) (700.0 mg, 2.12 mmol) in DCM (7.0 mL) using TFA (5.31g, 46.6 mmol) in DCM (10 mL) to afford 2-(4-amino-7-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (384f) (590.0 mg, 72% yield) TFA sait as an off white solid; *H NMR (300 MHz, DMSO-cfc) δ 8.52 (s, 1H), 8.47 - 8.29 (m, 3H), 7.70 (d, 10.6 Hz, 1H), 5.21 (s, 2H), 2.40 (d, J= 2.1 Hz, 3H).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-7-fluoro-6-methyl-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (384g)

Compound 384g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (384f) (75 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-2-azabicycIo[3.1.0]hexane-3-carboxamide (4a) (57.3 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (97 mg, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-7-fluoro-6

656 methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (384g) (50 mg, 62% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 10.76 (s, 1H), 8.58 (s, 1H), 8.55 (s, 2H, D₂O exchangeable), 8.46 (d, J= 7.3 Hz, 1H), 8.00 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.62 (d, J= 10.5 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.73 (d, J= 17.4 Hz, 1H), 5.37 (d, J= 17.3 Hz, 1H), 4.42 (dd, J= 9.0, 5.6 Hz, 1H), 3.92 - 3.83 (m, 1H), 2.39 (s, 3H), 2.37 - 2.14 (m, 2H), 1.98 - 1.84 (m, 1H), 1.13 - 0.97 (m, 1H), 0.85 - 0.74 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-cfc) δ -117.36; MS (ES+): 538.10 (M+l); (ES-): 536.10 (M-l).

Scheme 385

Préparation of (lR,3S,5R)-2-(2-(4-amino-7-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1,0]hexane-3-carboxamide (385a)

Compound 385a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-7-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (384f) (75 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (60 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (97 mg, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-7fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (385a) (75 mg, 82 % yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-îZ₆) δ 10.77 (s, 1H), 8.71 (s, 2H, D₂O exchangeable), 8.61 (s, 1H), 8.48 (d, J= 7.2 Hz, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.69 (t, J= 8.0 Hz, 1H), 7.62 (d, J = 10.4 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.69 (d, J= 17.4 Hz, 1H), 5.32 (d, J= 17.3 Hz, 1H), 4.37 (dd, J= 9.0, 5.9 Hz, 1H), 3.71-3.63 (m, 1H), 2.47-2.41 (m, 1H), 2.39 (s, 3H), 1.97

657 (dd, J= 13.3, 5.8 Hz, 1H), 1.30 (s, 3H), 1.06 - 0.89 (m, 2H); ¹⁹F NMR (282 MHz, DMSO-d₆) δ -116.70; MS (ES+): 552.10 (M+l); (ES-): 550.0 (M-l).

Scheme 386

Préparation of (lR,3S,5R)-2-(2-(4-amino-6,7-difluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (386g)

Step-1: Préparation ofN-(2-bromo-4,5-difluorophenyl)-2,2,2-trifluoroacetamide (386b)

Compound 386b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-4,5-difluoroaniline (386a)( 3.0 g, 14.70 mmol; CAS # 64695-79-0 ) in DCM 10 (30 mL) using trifluoroacetic acid anhydride (4.54 g, 21.62 mmol) and stirring at RT for 1 h.

This gave after workup N-(2-bromo-4,5-difluorophenyl)-2,2,2-trifluoroacetamide (386b) (3.5 g, 79% yield) as a dark brown solid; *H NMR (300 MHz, DMSO-î/ô) δ 11.43 (s, 1H), 8.11 7.98 (m, 1H), 7.78 (dd, J= 11.2, 7.9 Hz, 1H).

Step-2: Préparation of 2-amino-5,6-difluoro-lH-indole-3-carbonitriIe (386c)

Compound 386c was prepared according to the procedure reported in step-1 of scheme-11 and step-2 of scheme-374, from N-(2-bromo-4,5-difluorophenyl)-2,2,2-trifluoroacetamide

658 (386b) (4.5 g, 14.80 mmol) in DMSO (13.5 mL) using malononitrile (1.17 g, 17.71 mmol), L-proline (0.34 g, 2.95 mmol), Cul (0.28 g , 1.47 mmol), a solution of K2CO3 (4.09 g, 29.60 mmol) in water (13.5 mL) to afford 2-amino-5,6-difluoro-lH-indole-3-carbonitrile (386c) ( 1.4 g, 49% yield) as a white solid; Ή NMR (300 MHz, DMSO-J₆) δ 10.80 (s, 1H), 7.12 (ddd, J= 20.0, 10.9, 7.3 Hz, 2H), 6.93 (s, 2H).

Step-3: Préparation of 6,7-difhioro-9H-pyrimido[4,5-b]indol-4-amine (386d)

Compound 386d was prepared according to the procedure reported in step-1 of scheme-6 and step-3 of scheme-374, from 2-amino-5,6-difluoro-lH-indole-3-carbonitrile (386c) (1.4 g, 7.25 mmol) using trimethyl orthoformate (23.07 g, 217.43 mmol), AcOH (7 mL), NH₄OAc (2.79 g, 36.23 mmol) to afford 6,7-difluoro-9H-pyrimido[4,5-b]indol-4-amine (386d) (500 mg, 32% yield) as a brown solid.

Step-4: Préparation of ZerZ-butyl 2-(4-amino-6,7-difluoro-9H-pyrimido[4,5-b]indol-9yl)acetate (386e)

Compound 386e was prepared according to the procedure reported in step-2 of scheme-16 and step-4 of scheme-374, from 6,7-difluoro-9H-pyrimido[4,5-b]indol-4-amine (386d) (500 mg, 2.27 mmol) in DMF (6.5 mL) using ZerZ-butyl 2-bromoacetate (443.0 mg, 2.27 mmol), CS2CO3 (1.48 g, 4.54 mmol) to afford ZerZ-butyl 2-(4-amino-6,7-difluoro-9H-pyrimido[4,5b]indol-9-yl)acetate (386e) (220.0 mg, 29% yield ) as an off-white solid.

Step-5: Préparation of 2-(4-amino-6,7-difluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (386f)

Compound 386f was prepared according to the procedure reported in step-2 of scheme-1, from ZerZ-butyl 2-(4-arnino-6,7-difluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (386e) (500.0 mg, 1.50 mmol) in DCM (5.0 mL) using TFA (3.75 g, 32.90 mmol) in DCM (10 mL) to afford 2-(4-amino-6,7-difluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (386f) (360.0 mg, 61% yield) TFA sait as a white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 8.60 (dd, J= 11.5, 7.9 Hz, 1 H), 8.46 (s, 1H), 8.09 (s, 2H), 7.99 (dd, J= 11.3, 6.9 Hz, 1H), 5.20 (s, 2H).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6,7-difluoro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (386g)

Compound 386g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6,7-difluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (386f)

659 (76 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyndin-2-yl)2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (52 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (97 mg, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6,7-difluoro-9Hpyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicycIo[3.1.0]hexane-3carboxamide (386g) (57 mg, 64% yield) HCl sait as a white solid; 'H NMR (300 MHz, DMSO-J₆) δ 10.76 (s, 1H), 8.82 - 8.54 (m, 4H, 2H exchangeable), 8.06 - 7.87 (m, 2H), 7.70 (t, J= 7.9 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.76 (d, J= 17.4 Hz, 1H), 5.39 (d, J= 17.4 Hz, 1H), 4.42 (dd, J= 9.0, 5.5 Hz, 1H), 3.94 - 3.83 (m, 1H), 2.41 - 2.11 (m, 2H), 2.00 - 1.82 (m, 1H), 1.14 - 0.99 (m, 1H), 0.90 - 0.74 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-7₆) δ -138.00, 144.42; MS (ES+): 542.10 (M+l); (ES-): 540.0 (M-l).

Scheme 387

F

386f 387a

Préparation of (lR,3S,5R)-2-(2-(4-amino-6,7-difluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (387a)

Compound 387a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6,7-difluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (386f) (76 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (60 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (97 mg, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6,7-difluoro9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (387a) (55 mg, 60% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-7₆) δ 10.77 (s, 1H), 8.76 - 8.63 (m, 1H), 8.62 - 8.50 (m, 3H, 2H D₂O exchangeable), 8.01 (d, J= 8.2 Hz, 1H), 7.93 (dd, J= 11.2, 6.9 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.71 (d, J= 17.4 Hz, 1H), 5.34 (d, J= 17.3 Hz, 1H), 4.38

660 (t, J= 7.5 Hz, 1H), 3.71 - 3.62 (m, 1H), 2.51 - 2.39 (m, 1H), 1.99 (dd, J= 13.1, 5.8 Hz, 1H), 1.31 (s, 3H), 1.08 - 0.90 (m, 2H); ¹⁹FNMR (282 MHz, DMSO-î/ô) δ -138.26, -144.69; MS (ES+): 556.10 (M+l); (ES-): 554.0 (M-l).

Scheme 388

H

N. ^Br

HATU, DIPEA

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-9yI)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyçlo[3.1.0]hexane-3-carboxamide (388a)

Compound 388a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (383f) (78 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (59.9 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (0.131 mL, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4amino-6-fluoro-7-methoxy-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (388a) (40 mg, 47% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-î/ô) δ 10.71 (s, 1H), 8.65 (s, 2H, D₂O exchangeable), 8.58 (s, 1H), 8.50 (d, J= 11.9 Hz, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 7.51 (d, J= 7.4 Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.70 (d, J= 17.3 Hz, 1H), 5.42 (d, J= 17.3 Hz, 1H), 4.37 (t, J= 7.5 Hz, 1H), 3.96 (s, 3H), 3.72 - 3.66 (m, 1H), 2.52 - 2.42 (m, 1H), 2.00 (dd, J= 13.4, 5.9 Hz, 1H), 1.32 (s, 3H), 1.06 - 0.87 (m, 2H); ^I9F NMR (282 MHz, DMSO-î/ô) δ 139.92; MS (ES+): 568.10 (M+l); (ES-): 566.0 (M-l).

Scheme 389

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-7-fluoro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yI)-2-azabicyclo[3.1.0]hexane-3-carboxamide (389g)

Step-1: Préparation of N-(2,4-dibromo-5-fluorophenyl)-2,2,2-trifluoroacetamide (389b)

Compound 389b was prepared according to the procedure reported in step-1 of scheme-46, from 2,4-dibromo-5-fluoroaniline (389a)(5 g, 18.59 mmol; CAS # 1000578-04-0) in DCM (25 mL) using trifluoroacetic acid anhydride (5.86 g, 27.89 mmol) and stirring at RT for 1 h. This gave after workup N-(2,4-dibromo-5-fluorophenyl)-2,2,2-trifluoroacetamide (389b) ( 6.4 g, 94% yield) as an off-white solid; Ή NMR (300 MHz, DMSO-î/₆) δ 11.47 (s, 1 H), 8.22 (dd, J= 7.1, 2.1 Hz, 1H), 7.69 (dd, J= 9.2, 2.0 Hz, 1H).

Step-2: Préparation of 2-amino-5-bromo-6-fluoro-lH-indole-3-carbonitrile (389c)

Compound 389c was prepared according to the procedure reported in step-1 of scheme-11 and step-2 of scheme-374, from N-(2,4-dibromo-5-fluorophenyl)-2,2,2-trifluoroacetamide (389b) (6.4 g, 17.54 mmol) in DMSO (20 mL) using malononitrile (1.39 g, 21.046 mmol), L

662 proline (0.404 g, 3.51 mmol), Cul (0.334 g, 1.75 mmol), a solution of K2CO3 (3.647 g , 26.31 mmol) in DMW (6.4 mL) to afford 2-amino-5-bromo-6-fluoro-lH-indole-3-carbonitrile (389c) (1.7 g, 38% yield) as a yellow solid; Ή NMR (300 MHz, DMSO-ûfe) δ 10.93 (s, 1H), 7.27 (d, J= 6A Hz, 1H), 7.13 (d, J= 9.2 Hz, 1H), 7.01 (s, 2H).

Step-3: Préparation of 6-bromo-7-fluoro-9H-pyrimido[4,5-b]indol-4-amine (389d)

Compound 389d was prepared according to the procedure reported in step-1 of scheme-6 and step-3 of scheme-374, from 2-amino-5-bromo-6-fluoro-lH-indole-3-carbonitrile (389c) (1.8 g, 7.08 mmol) using trimethyl orthoformate (22.55 g, 212.55 mmol), AcOH (2.127 g, 35.425 mmol), NH4OAC (2.731 g, 35.425 mmol) to afford 6-bromo-7-fluoro-9H-pyrimido[4,5b]indol-4-amine (389d)(1.52 g, 76% yield) as a brown solid; *H NMR (300 MHz, DMSO-Jô) δ 12.12 (s, 1H), 8.70 (d, J= 6.8 Hz, 1H), 8.26 (s, 1H), 7.39 (d, J= 9.4 Hz, 1H), 7.34 (s, 2H).

Step-4: Préparation of tert-butyl 2-(4-amino-6-bromo-7-fluoro-9H-pyrimido[4,5-b]indol-9yl)acetate (389e)

Compound 389e was prepared according to the procedure reported in step-2 of scheme-16 and step-4 of scheme-374, from 6-bromo-7-fluoro-9H-pyrimido[4,5-b]indol-4-amine (389d) (1.5 g, 5.34 mmol) in DMF (60 mL) using tert-butyl 2-bromoacetate (1.15 g, 5.87 mmol), CS2CO3 (3.65 g, 11.21 mmol) to afford tert-butyl 2-(4-amino-6-bromo-7-fluoro-9Hpyrimido[4,5-b]indol-9-yl)acetate (389e) (1.5 g, 71% yield) as a brown solid; ’HNMR (300 MHz, DMSO-î/₆) δ 8.78 - 8.72 (m, 1H), 8.30 (d, J= 1.0 Hz, 1H), 7.82 - 7.76 (m, 1H), 7.46 (s, 2H), 5.12 (s, 2H), 1.42 - 1.37 (m, 9H).

Step-5: Préparation of 2-(4-amino-6-bromo-7-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (389f)

Compound 389f was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-6-bromo-7-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetate (389e) (1.5 g, 3.80 mmol) in DCM (24 mL) using TFA (9.52 g, 83.50 mmol) to afford 2-(4-amino-6bromo-7-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (389f) (1.8 g) TFA sait as an offwhite solid; 'HNMR (300 MHz, DMSO-îZ₆) δ 8.86 (d, J= 6.7 Hz, 1H), 8.48 (s, 1H), 8.18 (s, 2H), 7.95 (d, J= 9.8 Hz, 1H), 5.20 (s, 2H).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-7-fhioro-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (389g)

663

Compound 389g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-6-bromo-7-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (389f) (85 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (57.3 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (97 mg, 0.75 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-6-bromo-7fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (389g) (62 mg, 69% yield) HCl sait as a white solid; ’H NMR (300 MHz, DMSO-7₆) δ 10.76 (s, 1H), 8.88 (d, J= 6.7 Hz, 1H), 8.52 (s, 1H), 8.28 (s, 2H, D₂O exchangeable), 8.00 (d, J= 8.2 Hz, 1H), 7.83 (d, J= 9.7 Hz, 1H), 7.70 (t, J= 8.0

Hz, 1H), 7.32 (d, J= 7.7 Hz, 1H), 5.71 (d, 7= 17.4 Hz, 1H), 5.35 (d, 7= 17.4 Hz, 1H), 4.41 (dd, 7= 9.3, 5.5 Hz, 1H), 3.90 - 3.80 (m, 1H), 2.39-2.12 (m, 2H), 1.98- 1.84 (m, 1H), 1.11 - 0.98 (m, 1H), 0.87 - 0.75 (m, 1H); MS (ES+): 601.90 (M+l).

Scheme 390

₁₅ 389f ^{ΒΓ 390a}

Préparation of (lR,3S,5R)-2-(2-(4-amino-6-bromo-7-fluoro-9H-pyrimido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (390a)

Compound 390a was prepared according to the procedure reported in step-3 of scheme-1, 20 from TFA sait of 2-(4-amino-6-bromo-7-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (389f) (85 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (60 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (97 mg, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-625 bromo-7-fluoro-9H-pyrimido[4,5-b]indol-9-yl)acetyI)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (390a) (85 mg, 84% yield) HCl sait as a white solid;

664 'H NMR (300 MHz, DMSO-î/₆) δ 10.76 (s, 1H), 8.89 (d, J= 6.6 Hz, 1H), 8.53 (s, 1H), 8.35 (s, 2H, D₂O exchangeable), 8.01 (d, J= 8.2 Hz, 1H), 7.83 (d, J= 9.6 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.31 (d, J= 7.7 Hz, 1H), 5.68 (d, J= 17.4 Hz, 1H), 5.31 (d, J= 17.2 Hz, 1H), 4.42 4.27 (m, 1H), 3.68 - 3.61 (m, 1H), 2.49 - 2.40 (m, 1H), 1.99 - 1.89 (m, 1H), 1.30 (s, 3H),

1.04 - 0.90 (m, 2H); MS (ES+): 616.00 (M+l); (ES-): 613.85 (M-l).

Scheme 391

391a 391b 391c

^391d 391e

391f ^{Me 391g}

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-fluoro-6-rnethyl-9H-pyrimido[4,5-b]indol-910 yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (391g)

Step-1: Préparation of N-(2-bromo-6-fluoro-4-methylphenyl)-2,2,2-trifluoroacetamide (391b)

Compound 391b was prepared according to the procedure reported in step-1 of scheme-46, from 2-bromo-6-fluoro-4-methylaniline (391a) (5 g, 24.50 mmol; CAS # 18349-09-2) in DCM (25 mL) using trifluoroacetic acid anhydride (7.72 g, 36.76 mmol) in DCM (25 mL) and stirring at RT for 1 h. This gave after workup N-(2-bromo-6-fluoro-4-methylphenyl)21436

665

2,2,2-trifluoroacetamide (391b) (6.8 g, 93% yield) as a black solid; 'HNMR (300 MHz, DMSO-Je) δ 11.35 (s, 1H), 7.48 (s, 1H), 7.35 - 7.24 (m, 1H), 2.35 (s, 3H).

Step-2: Préparation of 2-amino-7-fluoro-5-methyl-lH-indole-3-carbonitrile (391c)

Compound 391c was prepared according to the procedure reported in step-1 of scheme-11 and step-2 of scheme-374, from N-(2-bromo-6-fluoro-4-methylphenyl)-2,2,2trifluoroacetamide (391b) (6.2 g, 20.66 mmol) in DMSO (19.3 mL) using malononitrile (1.64 g, 24.80 mmol), L-proline (0.476 g, 4.13 mmol), Cul (0.393 g, 2.07 mmol), a solution of K2CO3 (4.28 g, 30.99 mmol) in DMW (6.2 mL) to afford 2-amino-7-fluoro-5-methyl- 1Hindole-3-carbonitrile (391c) (1.55 g, 40% yield) as a brown solid; 'HNMR (300 MHz,

DMSO-î7₆) δ 11.16 (s, 1H), 6.77 (s, 1H), 6.68 (s, 2H), 6.63 - 6.53 (m, 1H), 2.31 (s, 3H).

Step-3: Préparation of 8-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-4-amine (391d)

Compound 391d was prepared according to the procedure reported in step-1 of scheme-6 and step-3 of scheme-374, from 2-amino-7-fluoro-5-methyl-lH-indole-3-carbonitrile (391c) (1.6 g, 8.46 mmol) using trimethyl orthoformate (26.92 g, 253.71 mmol), AcOH (2.54 g, 42.29 mmol), NH4OAC (3.26 g, 42.29 mmol) to afford 8-fluoro-6-methyl-9H-pyrimido[4,5-b]indol4-amine (391d) (1.45 g, 79% yield) as an off-white solid; 'H NMR (300 MHz, DMSO-dô) δ 12.19 (s, 1H), 8.26 (d, J= 1.0 Hz, 1H), 8.03 - 7.93 (m, 1H), 7.21 (s, 2H), 7.06 (d, J= 12.0 Hz, 1H), 2.46 (s, 3H).

Step-4: Préparation of tert-butyl 2-(4-amino-8-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-920 yl)acetate (391e)

Compound 391e was prepared according to the procedure reported in step-2 of scheme-16 and step-4 of scheme-374, from 8-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-4-amine (391d) (1.45 g, 6.71 mmol) in DMF (58 mL) using tert-butyl 2-bromoacetate (1.44 g, 7.38 mmol), CS2CO3 (4.588 g, 14.08 mmol) to afford tert-butyl 2-(4-amino-8-fluoro-6-methyl-9H25 pyrimido[4,5-b]indol-9-yl)acetate (391e) (1.6 g, 72% yield) as an off-white solid; 'H NMR (300 MHz, DMSO-îZô) δ 8.30 (s, 1H), 8.08 - 8.01 (m, 1H), 7.37 (s, 2H), 7.09 (d, J= 13.1 Hz, 1H), 5.12 (d, J= 1.8 Hz, 2H), 2.46 (s, 3H), 1.40 (s, 9H).

Step-5: Préparation of 2-(4-amino-8-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (391f)

666

Compound 391f was prepared according to the procedure reported in step-2 of scheme-1, from tert-butyl 2-(4-amino-8-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetate (391e) (1.6 g, 4.84 mmol) in DCM (25.6 mL) using TFA (12.15 g, 106.55 mmol) to afford 2-(4amino-8-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (391f) (1.85 g, 98% yield) TFA sait as a cream color solid; *H NMR (300 MHz, DMSO-cfc) δ 8.56 (d, J= 0.9 Hz, 1H), 8.46 (s, 2H), 8.15 (s, 1H), 7.23 (d, J= 13.3 Hz, 1H), 5.25 - 5.21 (m, 2H), 2.49 (s, 3H).

Step-6: Préparation of (lR,3S,5R)-2-(2-(4-amino-8-fluoro-6-methyl-9H-pyrimido[4,5b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2-azabicyclo[3.1,0]hexane-3-carboxamide (391g)

Compound 391g was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (391f) (75 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-2-azabicyclo[3.1.0]hexane-3-carboxamide (4a) (57.3 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (97 mg, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8-fluoro-6methyl-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-2azabicyclo[3.1.0]hexane-3-carboxamide (391g) (56 mg, 69% yield) HCl sait as a white solid; ‘H NMR (300 MHz, DMSO-J₆) δ 10.76 (s, 1H, D₂O exchangeable), 8.61 (s, 1H), 8.58 (s, 2H, D₂O exchangeable), 8.18 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 7.22 (d, 13.2 Hz, 1H), 5.77 (d, J= 17.4 Hz, 1H), 5.48 - 5.38 (m, 1H), 4.42 (dd, J= 9.1, 5.6 Hz, 1H), 3.91 - 3.81 (m, 1H), 2.48 (s, 3H), 2.39 - 2.14 (m, 2H), 2.02 - 1.83 (m, 1H), 1.16 - 0.97 (m, 1H), 0.68 - 0.53 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î7₆) δ 135.24; MS (ES+): 538.10 (M+l); (ES-): 536.10 (M-l).

Scheme 392

391f ^Me 392a

667

Préparation of (lR,3S,5R)-2-(2-(4-amino-8-fluoro-6-methyl-9H-pynmido[4,5-b]indol-9yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (392a)

Compound 392a was prepared according to the procedure reported in step-3 of scheme-1, from TFA sait of 2-(4-amino-8-fluoro-6-methyl-9H-pyrimido[4,5-b]indol-9-yl)acetic acid (391f) (75 mg, 0.15 mmol) in DMF (8 mL) using HCl sait of (lR,3S,5R)-N-(6-bromopyridin2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-carboxamide (8a) (59.9 mg, 0.180 mmol), HATU (114 mg, 0.300 mmol), DIPEA (97 mg, 0.750 mmol) and stirring at RT for 18 h. This gave after workup and purification as described in scheme-299, (lR,3S,5R)-2-(2-(4-amino-8fluoro-6-methyI-9H-pyrimido[4,5-b]indol-9-yl)acetyl)-N-(6-bromopyridin-2-yl)-5-methyl-2azabicyclo[3.1.0]hexane-3-carboxamide (392a) (31 mg, 37% yield) HCl sait as a white solid; Ή NMR (300 MHz, DMSO-t/₆) δ 10.76 (s, 1H, D₂O exchangeable), 8.59 (s, 1H), 8.51 (s, 2H, D₂O exchangeable), 8.17 (s, 1H), 8.01 (d, J= 8.2 Hz, 1H), 7.70 (t, J= 8.0 Hz, 1H), 7.31 (d, J = 7.7 Hz, 1H), 7.21 (d, J= 13.1 Hz, 1H), 5.72 (d, J= 17.5 Hz, 1H), 5.38 (d, J= 17.5 Hz, 1H), 4.37 (dd, J= 9.0, 5.9 Hz, 1H), 3.64 (dd, J= 5.5, 2.3 Hz, 1H), 2.49 - 2.42 (m, 4H), 1.98 (dd, J = 13.3, 5.9 Hz, 1H), 1.29 (s,3H), 1.06 - 0.97 (m, 1H), 0.83 - 0.74 (m, 1H); ¹⁹F NMR (282 MHz, DMSO-î/ô) δ -135.26; MS (ES+): 552.10 (M+l); (ES-): 549.95 (M-l).

Example 393

The ICso value of a compound (i.e., the concentration of the compound that inhibits 50% of the enzymatic activity) was calculated according to the procedure reported in US patent 6,653,340 Bl, e.g., column 74 (incorporated by reference).

Specifically, the compounds were dissolved in a stock solution of DMSO at 10.0 or 100 mM. A portion of this stock solution was added to assay buffer in a final volume of 50 pL. Controls included buffer alone and enzyme solutions to which DMSO was added. Substrate was added to the reaction wells immediately or after incubation at room température. The reaction rates were measured spectrophotometrically by the génération of product at 405 nm for 600 sec. Background absorbance at 690 nm was measured and subtracted from the absorbance at 405 nm for each well.

The reaction rate for enzyme alone was compared to the rate of enzyme in the presence of inhibitor and the percent inhibition was calculated as shown below:

Percent Inhibition = [Rate without inhibitor-Rate with inhibitor)/(Rate without inhibitor)] x 100

668

Factor D Esterolytic Assay:

An established esterolytic assay for the measurement of Factor D activity and inhibition of Factor D activity was used (Kam, C. M.; McRae, B. J.; Harper, J. W.; Niemann, M. A.; Volanakis, J. E.; Powers, J. C. Human complément proteins D, C2, and B Active site mapping with peptide thioester substrates. J Biol. Chem. 1987, 262, 3444-3451). For this assay Z-Lys-SBzl, 1.29 mM (Kim, S.; Narayana, S. V. L; Volanakis, J. E. Mutational analysis of the substrate binding site of human complément Factor D. Biochemistry. 1994, 33, 14393-14399.) was used as the substrate for Factor D (104 mM). Hydrolysis of this compound by Factor D liberated a free sulfhydryl group which is then reacted with 5,5'dithiobis(2nitrobenzoic acid) producing an intense yellow color (Habeeb, A. F. S. A. Reaction of protein sulfhydryl groups with Ellman's Reagent. Methods in Enzymol. 1976, 25, 457-464.). The assays were performed in 96 well microtiter plates and rates of hydrolysis were monitored at 405 nm on a Biotek Synergy H1 plate reader. Hydrolysis rates were reported as change in mOD/min. The assay was conducted in 100 mM HEPES, 500 mM NaCl, pH 7.5 containing 10% DMSO in a final volume of 50 pL per well.

An IC50, a compound concentration which inhibits 50% of the enzymatic activity, was calculated. Compounds in the examples were tested a minimum of three times. In the table below, three plus symbols (+++) are used to indicate compounds with an IC50 value of greater than lmicromolar (>1000 nM) concentration; Two (++) indicate compounds with an IC50 value between 1 and 0.1 micromolar (1000-100 nM) concentration; One (+) indicate compounds with an IC50 value less than 0.1 micromolar (<100 nM) concentration.

Table 1. Measured Ki (IC50) Value for Compounds. Three (+++) is used to dénoté compounds with an IC50 value of greater than lmicromolar (>1000 nM) concentration; Two (++) indicate compounds with an IC50 value between 1 and 0.1 micromolar (1000-100 nM) concentration; One (+) indicate compounds with an IC50 value less than 0.1 micromolar (<100 nM) concentration.

669

Compound	icso		Compound	icso		Compound	ic50
75d	+++	93a	++	106b	+++
76b	+++	94b	+++	112b	+
77b	+++	95c	+++	119a	++
78a	++	96e	+++	120c	+++
79a	++	97b	+++	121d	+++
80a	+	98b	++	122c	++
81a	+	99b	+++	123g	+++
82a	+++	100b	+++	118a	+++
83d	++	101b	++	117d	++
84a	++	102b	++	116b	++
85b	+++	103b	++	115e	++
86b	++	104a	+	114f	+
87f	+++	105b	++	113a	++
88f	+++	111b	++	66b	++
89b	++	107b	+++	124b	+++
90d	+++	108b	+++	68a	++
91a	+	109e	++	67c	++
92b	+++	HOd	+++	65a	+

670

Compound	icso		Compound	icso		Compound	ic50
64a	+	144g	++	151a	++
130e	++	145a	++	152d	++
131e	++	146a	+++	le	+
132d	+++	235a	+++	6e	+
133b	+	147a	+++	7b	+
20a	+	148d	++	21a	+
134b	+	19a	+	73a	+
135a	++	3b	+	74a	++
136a	+++	149b	++	153a	. +++
137b	+	4b	+	27a	+
140a	+++	5e	++	25a	+
141a	+++	150d	++	26a	+
138d	++	69a	++	8b	+
139b	+	70a	++	28a	+
125d	++	2d	+++	10b	+
126f	++	71a	++	llf	+
142d	+++	72a	++	12a	+
143d	++	9a	+	127d	+++

671

Compound	IC50		Compound	icso		Compound	ic50
128a	+++	50f	++	49c	+
154e	++	157a	++	58c	+
34e	+	158e	++	59c	+
13b	. ++	32f	+	17f	+
14a	+	43a	+	18a	+
15a	+	35f	+	31f	+
155f	+++	30f	+	60c	+
156e	++	47g	+	61a	+
45d	+	48d	+	129f	++
41f	+	38e	+	46g	++
37a	+	51g	+	62c	+
42a	+	52f	+	63d	+
16e	+	54g	+	53d	++
36e	+	44a	+	55a	+
39f	+	29f	+	56g	++
23f	+	22c	+++	57g	+
40a	+	22d	+++	159a	+
24a	+	33e	+	160a	+

672

Compound	icso		Compound	ic50		Compound	icso
171b	+	167a	+	204a	+
172a	+	168g	+	184g	+
173a	+	187c	+	185a	+
161a	+	188g	+	186a	+
174g	+++	195e	+	206c	++
175a	+++	198a	+	209b	+
176a	++	199a	+	210d	++
169f	+	200a	+	189g	+
177d	++	194a	+	193a	+
178d	++	201e	++	190a	+
162g	+	211e	++	191a	+
163g	+	212e	++	192g	+
164a	+	181a	+	196d	+
165a	+	182a	+	197c	+
170c	+	183a	+	214b	+
179d	+++	207b	+	213d	+
180e	++	208d	+	215g	+
166a	+	203a	+	216g	+

673

Compound	ICSO		Compound	ic50		Compound	icso
217a	+	224b	+++	271c	+
220a	++	225e	+	249a	+
202a	+	226c	+	252a	+
205c	+	219a	+	253a	+
221c	+	227c	+	254a	+
236f	+	228a	+	263c	+
237a	+	251f	++	272c	++
238g	+	233h	+	255c	+
239a	+	229c		256a	+
222a	+++	230a	+	273c	+ '
240g	+	231g	+	257g	+
241a	+	246b	+	258a	+
242g	+	247d	++	259d	++
243a	+	248d	+++	260a	+
244d	+	232a	+	261d	+
245a	+	234a	+	262a	+
218a	+	250b	+++	264a	+
223a	+	270a	+	265a	+

674

Compound	ic50
266b	+
267c	+
268b	+
269c	+
274g	+
275c	+
276g	+
277a	+
278g	+
279a	+
280g	+
281a	+
282a	+
283g	+
284a	+
285g	+
286a	+
287a	+

Compound	icso
288c	+
289a	+
290g	+
291a	+
292a	+
293a	++
293b	+
294c	+++
295a	+
296a	+
297d	+
298c	+
299a	+
300a	+
301a	+
302a	+
303a	+

675

Compound	IC50		Compound	IC5o		Compound	IC50
307c	+	313b	+	315a	+
304a	+	314a	+	316a	+
305a	+	312a	+	343a	+
308c	+	323a	+	344c	+
309a	+	324a	+	345c	+
310a	+	306a	+	346g	+
311a	+	325c	+	347a	+
317c	++	326c	+	338e	+
318c	+	327c	+	339g	+
319c	. +	328f	+	348c	+
320c	+	329a	+	349c	+
321d	+	330f	+	350a	+
322c	+	331a	+	351a	+
334a	+	336c	+	332c	+
335c	+	337a	+	333c	+

676

Compound	ic50		Compound	icso		Compound	ic50
365c	+	367g	+	389g	+
366c	+	368a	+	390a	+
352c	+	369g	+	376g	+
353a	+	362f	+	377a	+
354c	+	363a	. +	379h	+
355a	+	364d	+	380a	+
340a	+	364e	+	374g	+
341c	+	381g	+	375a	+
342a	+	382a	. +	391g	+
356c	+	385a	+	392a	+ .
357a	+	384g	+	370a	+
358c	++	386g	+	378c	+
359c	+	387a	+	371c	++
360c	+	383g	+	372a	++
361a	+	388a	+	373a	++

INCORPORATION BY REFERENCE

Ail of the U.S. patents and U.S. and PCT published patent applications cited herein 5 are hereby incorporated by reference.

EQUIVALENTS

The foregoing written spécification is considered to be sufficient to enable one skilled in the art to practice the invention. The présent invention is not to be limited in scope by examples provided, since the examples are intended as a single illustration of one aspect of 10 the invention and other functionally équivalent embodiments are within the scope of the invention. Various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and fall

677 within the scope of the appended claims. The advantages and objects of the invention are not necessarily encompassed by each embodiment of the invention.

Claims (15)

1. A compound represented by Formula (I), or a pharmaceutically acceptable sait

2. The compound of claim 1, wherein Y represents C(R^Y)2.

3. The compound of claim 1 or 2, wherein Y represents CH2.

4. The compound of any one of daims 1-3, wherein X represents a bond.

5 nocturnal hemoglobinuria, atypical hemolytic urémie syndrome, organ transplant rejection, myasthenia gravis, neuromyelitis optica, membranoproliferative glomerulonephritis, densedeposit disease, cold agglutinin disease, and catastrophic antiphospholipid syndrome.

54. The compound for use of claim 46, wherein the disease or condition characterized by aberrant complément System activity is selected from the group consisting of adult

5 25.

The compound of claim 15, wherein R² represents

26. The compound of claim 15, wherein R² represents ^r7Z

27. The compound of claim 26, wherein R² represents ^NH2

28.

The compound of claim 15, wherein R² represents

683

29.

The compound of claim 15, wherein R² represents

30.

The compound of claim 29, wherein R² represents

31. The compound of any one of daims 15-21, 24-26, 28, and 29, wherein R^7Z represents

-NR¹³R¹⁴.

5

32. The compound of any one of daims 15-21, 24-26, 28, and 29, wherein R^7Z represents

-NH₂.

33. The compound of any one of daims 15-20, wherein R^6Z represents -C(O)R¹³, -C(O)OR¹³, -C(O)NR¹³R¹⁴, or hydroxyalkyl.

34. The compound of any one of daims 15-21, 24-26, 28, and 29, wherein R^5Z represents 10 alkyl, halo, or -NR¹³R¹⁴.

35. The compound of any one of daims 15-20, 25, 28, and 29, wherein R^1Z represents CN, halo, haloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkyl, -C(O)R¹³, -SR¹³, -NR¹³R¹⁴, -OR¹³, -C(O)OR¹³, -C(O)NR¹³R¹⁴, or -NR¹³C(O)R¹⁴.

15

36. The compound of any one of daims 15-24, wherein each occurrence of R^2A independently represents-CN, -NO₂, halo, haloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl, -C(O)R¹³, -C(O)OR¹³, -NR¹³C(O)OR¹⁴, -SR¹³, -NR¹³R¹⁴, -OR¹³, -C(O)NR¹³R¹⁴, or -NR¹³C(O)R¹⁴.

20

37. The compound of any one of daims 1-36, wherein G is C(R³)₂.

38. The compound of any one of daims 1-37, having the structure of formula (la):

684

R² (la).

39. The compound of any one of daims 1-38, wherein two vicinal occurrences of R³ taken together with the carbon atoms to which they are bonded form an optionally substituted fused Ca-cycloalkyl.

5

40. The compound of any one of daims 1-39, wherein at least one occurrence of R³ is halo.

41. The compound of any one of daims 1-39, wherein at least one occurrence of R³ is fluoro.

42. The compound of any one of daims 1-39, wherein at least one occurrence of R³ is 10 methyl.

43. The compound of any one of daims 1-39, wherein two vicinal occurrences of R³ taken together with the carbon atoms to which they are bonded form an optionally substituted fused Ca-cycloalkyl and another occurrence of R³ is methyl.

44. The compound of daim 1, or a pharmaceutically acceptable sait thereof, selected from the 15 following table:

685

686

688

91a 7=\ Z—Z z // m )—t Z_x O \___/ ^O=\ zz O 22c and 22d 'Wï/ ^Me^o N-U'V--. flT) nh₂ ^Me (+) and (-) isomer

92b y । h Α^Νγ^Νγ^ΒΓ r—ζ O H₂N N N ⁰ NH_Z 33e ______ 11 .. Br ^O<1 f MV N=/ \r=V nh₂

93a F \ 1 ^{H n}^\<ⁿ^n _Br nh₂ 49c Rr ^rQÿ-C ^o<?S nV \==\ Λιη₂ ^cn

94b Jpy°^O ÇQ nh₂ 58c cû O Z v\ TZ (Ai y^o ks-Z A æ^zŒ V⁷ ° ⁷\ . ·

95c H ^<N Τ^ΝΊ A ° A N___N ί T/> Ν^ζΛ-Ν nh₂ 59c ____ 11 .. Br Aï? °^s <^z μΛ ⁿ=/ nh₂

689

691

693

123g x /^z=x y/ z z_x z^ M \ - ^z^V^m O \___/ ^o=\ ZI 4> O 176a V-o x° Ν~γ X^⁷ NH₂ '

118a 1; ΛΊ H ^N Τ^νΎ^νΎ^Βγ r4 o JL n ⁰ ço NH₂ Z==\ V .N ^Ni 169f Z'X- OY // A /V z'x-îX ^O Λ C° X

117d CyX Αθ ° ço NH₂ /==S\ V .N ^Ni 177d z^H=\ Z—Z t \s // M )----( A/V β J i (fi τι 1 ω J ZI ! z X-A ro “!

116b F, _p F Ôy’V rX o ⁰ ÇQ nh₂ ^Br 17 8d /^H=\ Z—G. Z z Ά // M )----( \ Z z²'-. P /Ύ Α^ζ/Ύ A ω / o=\ ZI / Z A# CD “1

115e F ? CVÙÔ <A Ο ÇQ ^NH2 0-^NH2 162g _Z-/A Λ, o=A ZI / Z Aa CD “Ί

696

132d 0 F H₃C N N ° ^ÇO nh₂ 188g H xi ^Br °Λ <^z μΛ N=Z \=A nh₂ ⁿ°²

133b ,^z=\ Z—z z // M >---< CPA_V ¹ O \ ? ^o=\ ZI b O 195e μι .. Br AVïJ ^o<l nV'v-n <^z ααΛ ^N \ nh₂

20a t? Zz-A \ €v i θ ^Xk-uTI ZI 198a tw ^N't\5 nh₂ ^x==/

134b ÇXyN^jÇX_c| rV^A ° ^F nJL? h T \ «n nh₂ /=/ > ia 199a Aï/' il 1 T \.___ / ^CF3 nh₂ —

135a ξ ÇXyN^jÇX_c| /A 0 F z'V'N ° Il Ί /> nh₂ Aa 200a Aï/ r^° < AY n A \A nh₂

697

699

ch₃

cf₃

700

146a u. 2 IZ r \ ( OJ ZA-i \=Z 186a z'-'W, . j· ^z~\ y\ — VAy j o=\ ZI 1 Z νΛ □□ “1

147a /—y o /^N^N ° O nh₂ 206c nA H / V-N N / Xk F \—N if- ^___N o o f y-^Br H₂N cf₃

148d F /VN ° ^f il T /> n^A<Z nh₂ ¹ 209b f=_z / N—t H / >>-N N F X—N F Vn 0 O J VBr ^NyJV ^XsV H₂N cf₃

19a F N—t ^H / >r-N . N / Vi 0 \—-N F VN 0 0 F yBr NyVJV. h₂n vA 210d N—X ^H / y-N N / vk V X—-N F V^N ο o F -yBr NxJL·^ \=^ H₂N 'îï-/ cf₃

3b F n4 h / ζ F~ y^N,_{x Rr} F V^{N 0} ° J ^F^b N^^/V 'XV h₂n Vv 189g ;;;y—\ H N^/⁸¹· [ o nvⁿv4 <^z XJ) NV V=\ nh₂ ^Me

704

705

25a V H D'à / \____/Ν^^Λ5=γ L 0 l^⁰ N-γ nh₂ ^CF3 240g wcr rv Ν—γ \=< / \ V-o nh₂ II ² 0

26a F N—\ H / \-N N ΛΛ U Y-N [ΓVn ^xo ο Γ \yBr n h₂n Χ-χ cf₃ 241g Ά* 1 °o^ _z N-X⁷ \===\ / \ \-0 nh₂ h ² 0

8b Aw Λ (\ n a v=x nh₂ ^Me 242a r? jT^^Z“X /X X<p z \ o zi ho / l ~ZOT —1

28a A NX ^===^ nh₂ ^CF3 243a Dû Z / / CM IZ O >o \ ___z^Z ζ/ΎΪ ^Z \ Λύ x⁷ \JL * yv^{z Z}Xi^Z -

10b A/ 1^° <\ N-jZ \=^\ nh₂ ^CH3 244d N=/M ΊμΠ V=7 nh₂

706

707

708

37a W 1^° 'A__/F V-OMe N^=/ \==F nh₂ 221c w 9% . nh₂ / H₃COOC

218a ΓΛΆ IVk^{Br N}\f A )> l o __,^Me nh₂ co₂h 250b /—\ H / \____/N^ A ⁰ Λί\ i ⁰ Az N^z^Nz^ f VÏA ^CN N=y NH₂

223a z% _X<A // Ο χ\_ y— z iF V-^ (f \ \\ /X. ^ZI o 2 / z s VL· co ----\ CD ^—I 270a ζΆ c'\ // y 0 XL·. XZ/ ~ Ca CO -n

224b Br N=/ >''-r>4^N^ ^-Ν Ό ^=0 N ^ZZ ⁿ=x A j N'A CO₂H nh₂ 271c ____ 11 .. Br SW ^QX| n^ⁿVf X7A nV NH₂ νχ^0Η₃

709

714

280g ?^z X²^ X Xao° y π \ zz / z CD “1 288c Z—(' z I \__/ M )-----( ^Ζ~~Λ |T o ZI / z ν-Λ tu -J

281a ? z Ζ-ΆμΧ s « X,z . α5ό° J~ -Π \ ZI ! z Υ-Λ tu -n 289a Ζ^!ϊ/ Æ° f X-O ^N λ nh₂ y^_N

282a w Γ o iXle ν_ύΜ eX-O \ih₂ ^oh 290g Æk o>? tu O^:=:\ ZI ! z kkZ CD “1

283g kx iXe ΝύΜ (' yJpi ^N=y X nh₂ 291a 'Wk¹ fk) Ι^^Νχ<\__,^Me Nip-'k'C nh₂ ^x==\ cf₃

716

717

310a Wf Me _Me ^N T\5 nh₂ ξ \ N N =ζ Me 311a Me ^^ΝχϊΓ^Ν\^/^Μθ ^N t\5 NH₂ \ N N Me

317c ώ z / LL XZ VsO Γ ^m Λ /? Lz^ XjT -/ ^~z O I z y— z \=z 318c /^H=\ z—(_x z I V // ro ) ( /Γ j ^ω o=\ zx / z Q

319c ,^H=\ z—(\ ^z X \\ + 60 >------< 2. _>Λ\- /Γ ] /^Z⁷^ o^z )> ω J o=\ zx m z ! -z. Q 320c tw liVw ^N T\> nh₂ cf₃

321d F, Wïf ï Q N /^° H Ί V-/ ^N T\) nh₂ ^x===\ cf₃ 322c w /^° H 1 V/ ⁿt\> nh₂ cf₃

718

334a N—N H oA ^Ny-N ) ⁰ Γ V h₂n ocf₃ 335c >:::r\ WW J O °η NW^NVw <⁷ ID nh₂ xy F

313b z—(' ² I \_/ ^w )----\ /Γ ] O V ω φ J o=\ ZI z ^zw g ” CD 314a L°ⁿ ^^Ny-N _Me ⁿt\5 nh₂ Me

312a z—(^z z X \_/ K> )------( A/^Z> .J _ γύ y^n jry^yo y. / z VA CD 323a w l^o nwⁿ-w4 N-y Vy NH₂ ^CF3 m Va z / xz yo ω-\ /9 wA CO z / xz y⁰ y

324a wM W N-W NH₂ ^cfs 306a CO LL O ~\A * yy ^ZXi^²

719

325c w A⁰ <\ XT? N-/ \=X nh₂ ^CF3 326c w 1 O ^N A⁰ λ-Γι Ν-γ X=\ nh₂ ^CF3

327c Wtf 1 Q N i^⁰ < M? Ny \=\ NH₂ ^CF3 328f [^° <x fl-TY⁰ N'Y nh₂

329a Ύ A⁰ λΑΥ° NA \=X nh₂ 330f Wï/ f^⁰ n^^NV-x N~Y X^x NH₂ /°

331a Yx 1^° NA NH₂ θ 336c ⁿA Kl ο=ζ 'Y y-N > ° Γυ^βγ Y y-N W h₂n VA O]

337a W °A fin N=/ \^( NH₂ YX F 315a ’Y-if fAle nVvY f tn N Y NH₂ F% nY Me

721

350a ^rtW Ion X VL_,^Me 1 ^N nh₂ '—( cf₃ 351a ’Xü” r\.° f X-X.N N^/ \===\ nh₂ ⁰¹⁼³

332c W 1^° « M/ nh₂ ^CF3 333c F> V X r? oW XX j¹ o rW Tl z ^ω VA / tx> τι -¹

365c H M 7^CI îxx w° M? N -X X=\ NH₂ ^CF3 366c μ ' κι Br XX x° NX^NvL N-X \=\ nh₂ ^CF3

352c _/?X Z—V -Z. z \_/ fo )------( W ZI ! Z. VjZ CD Π 353a Xi/ yj^⁰ ^>^Nx_-N__Me nh₂ Xo

723

359c ΥΊΓΑ_.^{Me n}tv5 nh₂ y V 360c z—<^z z I \_/ M )----( ^y~^zX] o ^v CD J . ZI Γ z y_j7 tu ^—I

361a /Ά_ο YVX_ Me ^N TW nh₂ 367g ^ry^s··© l⁰ n^ⁿv^ / mY° N--Z \^\ NH₂ ^CF3

368a /^Z=x Z—<\ Z I Ά // ΙΌ \-----< ^Z~~a Z 1 /fr—Y~” S A 'A ω i j o=\ ^x ZI ! z tu “1 369g w©r rS NZ^NU N-Z \=\ nh₂ ^CF3

362f W©·' a.° Z© < y_fS N=/ ^X _N=\ nh₂ ^CF3 363a tv©·' Γ o Awie nV'vY <^z M7 N^=/ ^X _N=\ . nh₂ ^CF3

364e y©:. ille f X-O N© NH₂ ^CF3 364d <ÿX [H/le nVvY <^z M? N==/ \=\ nh₂ ^CF3

724

381g N—t ^H 0=^ Ίτ V_N > ° Γν r y-N w h₂n F 382a i? Z—, Ζ-Λ \ Z /Y ° zx CD ^—1

385a N—C H o=( ^Y^Ny-^N > ° fr^{r N}<sJ<A> h₂n xA-f 384g m TZ \ LL ΛΎ ^O r—Z Yyx'W' o A/ z \ 'x y~-z X

386g I -, N> --. Λ \ Z /Ύ ° _: / ?^z T! \ ZI CD t 387a T -, ro —. Λ \ Z /Y ° T| \ XX CD —J

383g J h,,___ N—\ H o=( ^Y^Ny-N .n > ° fy^Br NXM. h₂n γΚο⁷ F 388a \ Z /Ύ ° - ' ^x °-=s< o \ \ zx CD -1

389g N-i H o=( Y'^Ny-N > ° fr^r y-N W H₂N Vïï-z'F Br 390a N—X H yⁿ _Vn ,n > ⁰ fy^Br © })~Ν W h₂n Br

725

376g N—χ H o=/ ^^^Ny-N > ° ΓΓ r y-n \=y h₂n XX Br 377a I _z A \ Z /Y ° CCJ^\ _-/ \ ¹ ZI ç 00 —1

379h nX H 0=/ ^V^Ny-N > ° Γ V y-N W H₂N XXX Br 380a ^NX N °=( y-n > ° Γ V γ-N W H₂N Br

374g nX H o=( ^V'Ny-N > ° fr^r y-N W ⁿ.jy h₂n X₅Xo^/ Br 375a ^^hJL ^NX N o=( X y,n ) ° Γ V h₂n XXo^/ Br

391g ta Va Z J / u—/y ^TZ v w v^o <\ γ\λ ^χ z—x y ° 392a + XVr? y⁷ W /K ^{zx 71} / 1 Z νΛ CD —r

370a w °Λ _Ν^^ΝνΧ f XJ? N=/ \=< \ \-0 NH₂ F \ 378c // z O /CLr? jT/Z-Ζ jCX-- γ¹ Ca 00 1

726

45. A pharmaceutical composition, comprisingthe compound of any one of the preceding daims, or a pharmaceutically acceptable sait thereof and a pharmaceutically acceptable carrier.

46. A compound of any one of daims 1-44, or a pharmaceutically acceptable sait thereof 5 for use in a method of treating or preventing a disease or condition characterized by aberrant complément System activity.

47. The compound for use of claim 46, wherein the disease or condition characterized by aberrant complément System activity is an immunological disorder.

48. The compound for use of claim 46, wherein the disease or condition characterized by 10 aberrant complément System activity is a disease of the central nervous System.

49. The compound for use of claim 46, wherein the disease or condition characterized by aberrant complément System activity is a neurodegenerative disease or neurological disease.

50. The compound for use of claim 46, wherein the disease or condition characterized by 15 aberrant complément System activity is a rénal disease.

51. The compound for use of claim 46, wherein the disease or condition characterized by aberrant complément System activity is a cardiovascular disease.

727

52. The compound for use of claim 46, wherein the disease or condition characterized by aberrant complément System activity is a cardiometabolic disease.

53. The compound for use of claim 46, wherein the disease or condition characterized by aberrant complément System activity is selected from the group consisting of paroxysmal

5. The compound of any one of daims 1-3, wherein X represents CH2.

5 thereof:

R² (I);

wherein, independently for each occurrence:

X is a bond or C(R^x)2j

Y is a bond, C(R^Y)2, or -N(R^b)

G is S or C(R³)₇;

R^a and R^b are each independently H or (Ci-Ce)alkyl;

R¹ represents optionally substituted aryl, heteroaryl, alkyl, cycloalkyl, alkenyl, or cycloalkenyl;

R² represents optionally substituted bicyclic ortricyclic heteroaryl;

R³ is independently for each occurrence H, halogen, -CN, -NH2, -CH2NH2, (CiCe)alkoxy or (Ci-Ce)alkyl; or two vicinal occurrences of R³ taken together with the carbon atoms to which they are bonded form an optionally substituted fused (C3-C7)cycloalkyl or (C6)aryl; or two geminal occurrences of R³ taken together with the carbon atom to which they are bonded form an optionally substituted spiro (C3-C7)cycloalkyl; or two hominal occurrences of R³ taken together with the carbon atoms to which they are bonded form an optionally substituted bridged (C3-C₇)cycloalkyl;

R^x is independently for each occurrence H, (Ci-Ce)alkyl, or (C3-C₇)cycloalkyl;

679

R^Y is independently for each occurrence H, (Ci-Ce)aIkyl, or (C3-C?)cycloalkyl;

or an occurrence of R^Y and a substituent on R² taken together with the intervening atoms form a ring;

optional substituents on R¹ or R² each independently represent halogen, -CN, —NO2, -OR¹³, -NR¹³R¹⁴, -C(O)R¹³, -C(O)OR¹³, -C(O)NR¹³R¹⁴, -OC(O)R¹³, -NR¹³C(O)R¹⁴, -OC(O)NR¹³R¹⁴, -OC(O)OR¹³, -NR¹³C(O)OR¹⁴, -NR¹³C(O)NR¹³R¹⁴, —OS(O)P(R¹³), -SR¹³,-NR¹³S(O)_P(R¹⁴), or optionally substituted alkyl, alkenyl, alkynyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cycloalkoxyalkyl, aryloxyalkyl, aralkyl, heteroaralkyl, heteroaryl, aryl, aryloxy, cycloaIkyl, (cycloalkyl)alkyl, heterocycloalkyl, or (heterocycloalkyl)alkyl;

or wherein two substituents on R¹ or two substituents on R², taken together with the intervening atoms, form a ring;

R¹³ and R¹⁴, independently for each occurrence, represent H or optionally substituted àlkyl, haloalkyl, alkenyl, alkynyl, aryl, or heteroaryl; and p is 0,1, or 2.

6. The compound of any one of daims 1-5, wherein R¹ represents optionally substituted aryl or heteroaryl.

7. The compound of any one of daims 1-5, wherein R¹ represents optionally substituted heteroaryl.

8. The compound of any one of daims 1-5, wherein R¹ represents optionally substituted phenyl (e.g., 3-halophenyl, or 2,3-dihalophenyl), (Cs-Cejcycloalkyl, alkenyl, pyridinyl (e.g., 6-halopyridin-2-yl), or pyrazinyl (e.g., 6-halopyrazin-2-yl).

9. The compound of any one of daims 1-8, wherein R¹ is mono-, di-, or tri- substituted.

680

10 ischemia/reperfusion injury, acute kidney injury, and organ transplantation, e.g., kidney transplant, systemic inflàmmatory response syndrome, septic shock, trauma, cancer, antibody-mediated rejection, Berger's disease, delayed graft function, granulomatosis with polyangiitis, graft versus host disease, hematopoietic stem cell transplant-related thrombotic microangiopathy, immune complex-mediated membranoproliferative

10 respiratory distress syndrome, myocardial infarct, lung inflammation, sepsis, cardiopulmonary bypass, burns, asthma, restenosis, multiple organ dysfunction, GuillainBarré syndrome, hémorrhagie shock, glomerulonephritis, systemic lupus erythematosus, rheumatoid arthritis, infertility, Alzheimer's disease, multiple sclerosis, platelet storage, and hemodialysis.

15

55. The compound for use of claim 46, wherein the disease or condition characterized by aberrant complément System activity is selected from the group consisting of antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), warm autoimmune hemolytic anémia, IgA nephropathy, C3 glomerulonephritis, and focal segmentai glomerulosclerosis.

56. The compound for use of claim 46, wherein the disease or condition characterized by 20 aberrant complément System activity is a hematological disorder.

57. The compound for use of claim 46, wherein the disease or condition characterized by aberrant complément System activity is an ocular disorder or an eye disorder.

58. The compound for use of claim 46, wherein the disease or condition characterized by aberrant complément System activity is macular degeneration, age-related macular

25 degeneration (AMD), wet AMD, géographie atrophy, macular edema, diabetic macular edema, choroidal neovascularization (CNV), uveitis, Behcet's uveitis, proliférative diabetic retinopathy, non-proliferative diabetic retinopathy, glaucoma, hypertensive retinopathy, a corneal neovascularization disease, post-corneal transplant rejection, a corneal dystrophie

728 disease, an autoimmune dry eye disease, Stevens-Johnson syndrome, Sjogren's syndrome, an environmental dry eye disease, Fuchs' endothélial dystrophy, retinal vein occlusion, or post-operative inflammation.

59. The compound for use of claim 46, wherein the disease or condition characterized by 5 aberrant complément System activity is selected from the group consisting of obesity, insulin résistance, diabètes, dyslipidemia, nephropathy, and neuropathy.

60. The compound for use of claim 46, wherein the disease or condition is angioedema.

61. The compound for use of claim 46, wherein the disease or condition is hereditary angioedema.

10

62. The compound for use of claim 46, wherein the disease or condition is acquired angioedema.

63. The compound for use of claim 46, wherein the disease or condition is selected from the group consisting of antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), paroxysmal nocturnal hemoglobinuria, and thrombotic microangiopathy.

15

64. The compound for use of claim 46, wherein the disease or condition is selected from the group consisting of Alzheimer's disease, multiple sclerosis, neuromyelitis optica, generalized myasthenia gravis, Guillain-Barré syndrome, Parkinson's disease, and schizophrenia.

65. The compound for use of claim 46, wherein the disease or condition is periodontitis.

20

66. The compound for use of claim 46, wherein the disease or condition is Crohn's disease.

67. The compound for use of claim 46, wherein the disease or condition is selected from the group consisting of asthma, chrohic obstructive pulmonary disease, and acute respiratory distress syndrome.

25

68. The compound for use of claim 46, wherein the disease or condition is atheroscerosis.

L

729

69. The compound for use of claim 46, wherein the disease or condition is selected from the group consisting of age-related macular degeneration (AMD), uveitis, glaucoma, and wet AMD.

70. The compound for use of claim 46, wherein the disease or condition is myocardial infarction.

71. The compound for use of claim 46, wherein the disease or condition is selected from the group consisting of atypical hemolytic urémie syndrome, C3 glomerulopathy, lupus nephritis, IgA nephropathy, and membranous nephropathy.

72. The compound for use of claim 46, wherein the disease or condition is selected from the group consisting of rheumatoid arthritis, osteoarthritis, bullous pemphigoid, psoriasis, hidradenitis suppurativa, and burns.

73. The compound for use of claim 46, wherein the disease or condition is hemodialysis.

74. The compound for use of claim 46, wherein the disease or condition is selected from the group consisting of rheumatoid arthritis, osteoarthritis, bullous pemphigoid, psoriasis, hidradenitis suppurativa, and burns.

75. The compound for use of claim 46, wherein the disease or condition is selected from the group consisting of ischemia/reperfusion injury, acute kidney injury, and organ transplantation, e.g., kidney transplant.

76. The compound for use of claim 46, wherein the disease or condition is selected from the group consisting of systemic inflammatory response syndrome, sepsis, septic shock, trauma, systemic lupus erythematosus, hereditary angioedema, and cancer.

77. The compound for use of claim 46, wherein the disease or condition is selected from the group consisting of antibody-mediated rejection, antiphospholipid syndrome, Berger's disease, C3 glomerulonephritis, cold agglutinin disease, cardiopulmonary bypass, dense deposit disease, delayed graft function, géographie atrophy, granulomatosis with polyangiitis, graft versus host disease, hematopoietic stem cell transplant-related thrombotic microangiopathy, immune complex-mediated membranoproliferative

L

730

glomerulonephritis, immune-mediated necrotizing myopathy, idiopathic polypoidal choroidal vasculopathy, microscopie polyangitis, pyoderma gangrenosum, Stargardt Disease 1, and warm type autoimmune menolytic anémia.

78. The compound for use of claim 46, wherein the disease or condition characterized by 5 aberrant complément System activity is selected from the group consisting of obesity, insulin résistance, diabètes, dyslipidemia, nephropathy, neuropathy, angioedema, e.g., hereditary angioedema or acquired angioedema, thrombotic microangiopathy, Parkinson's disease, schizophrenia, periodontitis, Crohn's disease, C3 glomerulopathy, membranous nephropathy, osteoarthritis, bullous pemphigoid, psoriasis, hidradenitis suppurativa, of

10. The compound of any one of daims 1-7, wherein R¹ represents optionally substituted pyridinyl.

11. The compound of any one of daims 1-7 and 10, wherein R¹ represents optionally substituted 2-pyridinyl.

5

12. The compound of any one of daims 1-7,10, and 11, wherein R¹ represents

13. The compound of any one of daims 1-6, wherein R¹ represents optionally substituted phenyl.

F

W¹

14. The compound of any one of daims 1-6 and 13, wherein R¹ represents

10

15. The compound of any one of daims 1-14, wherein R² represents

Z¹ represents N or CR^1Z;

Z² represents N or CR^2Z;

Z³ represents N or C;

681

Z⁴ represents N or CR^4Z;

Z⁵ represents N or CR⁵,

Z⁶ represents N or CR^6Z;

Z⁷ represents N or CR^7Z;

Z⁸ represents C;

Z⁹ represents N or C;

k is an integer from 1-4;

m is an integer from 1-3; and each occurrence of R^1Z, R^2Z, R^4Z, R^5Z, R^6Z, R^7Z, R^2A independently represents H, halogen, -CN, -NO2, -OR¹³, -NR¹³R¹⁴, -C(O)R¹³, -C(O)OR¹³, -C(O)NR¹³R¹⁴, -OC(O)R¹³, -NR¹³C(O)R¹⁴, -OC(O)NR¹³R¹⁴, -OC(O)OR¹³, -NR¹³C(O)OR¹⁴, -NR¹³C(O)NR¹³R¹⁴, -OS(O)p(R¹³), -SR¹³, -NR¹³S(O)P(R¹⁴), or optionally substituted alkyl, alkenyl, alkynyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cycloalkoxyalkyl, aryloxyalkyl, aralkyl, heteroaralkyl, heteroaryl, aryl, aryloxy, cycloalkyl, (cycloalkyl)alkyl, heterocycloalkyl, or (heterocycloalkyl)alkyl; or wherein an occurrence of R^6Z and an occurrence of R^7Z taken together with the intervening atoms form a ring; or wherein an occurrence of R^Y and an occurrence of R^2Z taken together with the intervening atoms form a ring.

16. The compound of claim 15, wherein Z², Z³, Z⁴, and Z⁶ represent N.

17. The compound of claim 15, wherein Z³, Z⁴, and Z⁶ represent N.

18. The compound of claim 15, wherein Z⁹, Z⁴, and Z⁶ represent N.

19. The compound of claim 15, wherein Z³ and Z⁴ represent N.

20. The compound of claim 15, wherein Z¹, Z³, Z⁴, and Z⁶ represent N.

682

21.

The compound of claim 15, wherein R² represents

22.

The compound of claim 21, wherein R² represents

23. The compound of claim 21 or 22, wherein k represents 2.

24.

The compound of claim 15, wherein R² represents

15 glomerulonephritis, immune-mediated necrotizing myopathy, idiopathic polypoidal choroidal vasculopathy, microscopie polyangitis, pyoderma gangrenosum, and Stargardt Disease 1.