Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
  • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
  • C07D487/08—Bridged systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
  • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
  • A61K31/5375—1,4-Oxazines, e.g. morpholine
  • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
  • A61K31/5375—1,4-Oxazines, e.g. morpholine
  • A61K31/5386—1,4-Oxazines, e.g. morpholine spiro-condensed or forming part of bridged ring systems
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
  • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
  • C07D491/10—Spiro-condensed systems
  • C07D491/107—Spiro-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
  • C07D498/08—Bridged systems

Definitions

• KRAS G12D INHIBITORS FIELD OF THE INVENTION relates to compounds that inhibit KRas G12D.
• the present invention relates to compounds that inhibit the activity of KRas G12D, pharmaceutical compositions comprising the compounds and methods of use therefor.
• KRas Kirsten Rat Sarcoma 2 Viral Oncogene Homolog
• KRas is a small GTPase and a member of the Ras family of oncogenes.
• KRas serves as a molecular switch cycling between inactive (GDP-bound) and active (GTP-bound) states to transduce upstream cellular signals received from multiple tyrosine kinases to downstream effectors to regulate a wide variety of processes, including cellular proliferation (e.g., see Alamgeer et al., (2013) Current Opin Pharmcol.13:394-401). [0003] The role of activated KRas in malignancy was observed over thirty years ago (e.g., see Santos et al., (1984) Science 223:661-664).
• KRas Aberrant expression of KRas accounts for up to 20% of all cancers and oncogenic KRas mutations that stabilize GTP binding and lead to constitutive activation of KRas and downstream signaling have been reported in 25 -30% of lung adenocarcinomas.
• Single nucleotide substitutions that result in missense mutations at codons 12 and 13 of the KRas primary amino acid sequence comprise approximately 40% of these KRas driver mutations in lung adenocarcinoma.
• KRAS G12D mutation is present in 25.0% of all pancreatic ductal adenocarcinoma patients, 13.3% of all colorectal carcinoma patients, 10.1% of all rectal carcinoma patients, 4.1% of all non-small cell lung carcinoma patients and 1.7% of all small cell lung carcinoma patients (e.g., see The AACR Project GENIE Consortium, (2017) Cancer Discovery;7(8):818-831. Dataset Version 4).
• KRas The well-known role of KRas in malignancy and the discovery of these frequent mutations in KRas in various tumor types made KRas a highly attractive target of the pharmaceutical industry for cancer therapy.
• KRas G12D inhibitors of KRas, particularly inhibitors of activating KRas mutants, especially KRas G12D.
• compositions comprising a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.
• methods for inhibiting KRas G12D activity in a in a cell comprising contacting the cell with a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof as defined herein.
• the contacting is in vitro. In one embodiment, the contacting is in vivo.
• Also provided herein is a method of inhibiting cell proliferation, in vitro or in vivo, the method comprising contacting a cell with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof as defined herein.
• methods for treating cancer in a patient comprising administering a therapeutically effective amount of a compound or pharmaceutical composition of the present invention or a pharmaceutically acceptable salt thereof to a patient in need thereof.
• Also provided herein is a method of treating a KRas G12D-associated disease or disorder in a patient in need of such treatment, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof as defined herein.
• a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof as defined herein for use in therapy is also provided herein.
• a compound of Formula (I), or a pharmaceutically acceptable salt thereof for use in the inhibition of KRas G12D.
• a compound of Formula (I), or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof as defined herein for use in the treatment of a KRas G12D-associated disease or disorder.
• a use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, as defined herein in the manufacture of a medicament for the inhibition of activity of KRas G12D is also provided herein.
• a compound of Formula (I), or a pharmaceutically acceptable salt thereof, as defined herein, in the manufacture of a medicament for the treatment of a KRas G12D-associated disease or disorder is also provided herein.
• Also provided herein is a method for treating cancer in a patient in need thereof, the method comprising (a) determining that the cancer is associated with a KRas G12D mutation (i.e., a KRas G12D-associated cancer); and (b) administering to the patient a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
• a process for preparing a compound of Formula (I), or a pharmaceutically acceptable salt thereof is also provided herein.
• a compound of Formula (I), or a pharmaceutically acceptable salt thereof obtained by a process of preparing the compound as defined herein.
• the present invention relates to inhibitors of KRas G12D.
• the present invention relates to compounds that inhibit the activity of KRas G12D, pharmaceutical compositions comprising a therapeutically effective amount of the compounds and methods of use therefor.
• DEFINITIONS [0026] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this invention belongs. All patents, patent applications, and publications referred to herein are incorporated by reference.
• KRas G12D refers to a mutant form of a mammalian KRas protein that contains an amino acid substitution of an aspartic acid for a glycine at amino acid position 12.
• the assignment of amino acid codon and residue positions for human KRas is based on the amino acid sequence identified by UniProtKB/Swiss-Prot P01116: Variantp.Gly12Asp.
• a “KRas G12D inhibitor” refers to compounds of the present invention that are represented by Formula (I), as described herein. These compounds are capable of negatively modulating or inhibiting all or a portion of the enzymatic activity of KRas G12D.
• KRas G12D-associated disease or disorder refers to diseases or disorders associated with or mediated by or having a KRas G12D mutation.
• a non-limiting example of a KRas G12D-associated disease or disorder is a KRas G12D-associated cancer.
• the term “subject,” “individual,” or “patient,” used interchangeably, refers to any animal, including mammals such as mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, primates, and humans. In some embodiments, the patient is a human.
• the subject has experienced and/or exhibited at least one symptom of the disease or disorder to be treated and/or prevented.
• the subject has been identified or diagnosed as having a cancer having a KRas G12D mutation (e.g., as determined using a regulatory agency-approved, e.g., FDA-approved, assay or kit).
• the subject has a tumor that is positive for a KRas G12D mutation (e.g., as determined using a regulatory agency-approved assay or kit).
• the subject can be a subject with a tumor(s) that is positive for a KRas G12D mutation (e.g., identified as positive using a regulatory agency- approved, e.g., FDA-approved, assay or kit).
• the subject can be a subject whose tumors have a KRas G12D mutation (e.g., where the tumor is identified as such using a regulatory agency- approved, e.g., FDA-approved, kit or assay).
• the subject is suspected of having a KRas G12D gene-associated cancer.
• an assay is used to determine whether the patient has KRas G12D mutation using a sample (e.g., a biological sample or a biopsy sample (e.g., a paraffin-embedded biopsy sample) from a patient (e.g., a patient suspected of having a KRas G12D-associated cancer, a patient having one or more symptoms of a KRas G12D-associated cancer, and/or a patient that has an increased risk of developing a KRas G12D-associated cancer) can include, for example, next generation sequencing, immunohistochemistry, fluorescence microscopy, break apart FISH analysis, Southern blotting, Western blotting, FACS analysis, Northern blotting
• the assays are typically performed, e.g., with at least one labelled nucleic acid probe or at least one labelled antibody or antigen-binding fragment thereof.
• regulatory agency is a country’s agency for the approval of the medical use of pharmaceutical agents with the country.
• a non-limiting example of a regulatory agency is the U.S. Food and Drug Administration (FDA).
• FDA U.S. Food and Drug Administration
• acyl refers to -C(O)CH 3 .
• C1-C6 alkyl refers to straight and branched chain aliphatic groups having from 1-6 carbon atoms, or 1-4 carbon atoms, or 1-3 carbon atoms, respectively.
• alkyl groups include, without limitation, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl.
• C1-C3 haloalkyl and C1-C4 haloalkyl refer to a C1-C3 alkyl chain or C1- C4 alkyl chain, respectively, as defined herein in which one or more hydrogen has been replaced by a halogen. Examples include trifluoromethyl, difluoromethyl and fluoromethyl.
• An "C1-C4 alkylene,” group is a C1-C4 alkyl group, as defined hereinabove, that is positioned between and serves to connect two other chemical groups. Exemplary alkylene groups include, without limitation, methylene, ethylene, propylene, and butylene.
• C1-C3 alkoxy and “C1 – C4 alkoxy” refer to –OC1 – C3 alkyl and -OC1-C4 alkyl, respectively, wherein the alkyl portion is as defined herein above.
• cycloalkyl as employed herein includes saturated and partially unsaturated cyclic hydrocarbon groups having 3 to 12 carbons, for example 3 to 8 carbons, and as a further example 3 to 6 carbons, wherein the cycloalkyl group additionally is optionally substituted with one or more R 6 groups as defined herein.
• cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.
• the term “cycloalkyl” also includes bridged cycloalkyls, such as bicyclo[1.1.1]pentanyl. [0039]
• the terms “C1-C3 hydroxyalkyl” and “C1-C4 hydroxyalkyl” refer to –C1- C3 alkylene-OH and -C1-C4 alkylene-OH, respectively.
• C2-C4 hydroxyalkynyl refers to -C2-C4 alkynylene-OH.
• An "aryl” group is a C 6 -C 14 aromatic moiety comprising one to three aromatic rings, which is optionally substituted with one or more R 6 or with one or more R 7 as defined herein.
• the aryl group is a C6-C10 aryl group. Examples of aryl groups include, without limitation, phenyl, naphthyl, anthracenyl, fluorenyl, and dihydrobenzofuranyl.
• Aryl also refers to bicyclic or tricyclic ring systems in which one or two rings, respectively, of said aryl ring system may be saturated or partially saturated, and wherein if said ring system includes two saturated rings, said saturated rings may be fused or spirocyclic.
• An example of an aryl ring system comprising two saturated rings wherein the rings are spirocyclic includes the following ring system: .
• An "araC1-C6 alkyl" or "arylalkyl” group comprises an aryl group covalently linked to an alkyl group, either of which may independently be optionally substituted or unsubstituted.
• An example of an aralkyl group is (C6-C10)aryl(C1- C6)alkyl-, including, without limitation, benzyl, phenethyl, and naphthylmethyl.
• An example of a substituted araC1-C6 alkyl is wherein the alkyl group is substituted with hydroxyalkyl.
• a “heterocyclyl” or “heterocyclic” group is a ring structure having from 3 to 12 atoms, for example 4 to 8 atoms, wherein one or more atoms are selected from the group consisting of N, O, and S wherein the ring N atom may be oxidized to N-O, and the ring S atom may be oxidized to SO or SO 2 , the remainder of the ring atoms being carbon.
• the heterocyclyl may be a monocyclic, a bicyclic, a spirocyclic or a bridged ring system.
• the heterocyclic group is optionally substituted with one or more R 6 on ring carbon or ring nitrogen at one or more positions, wherein R 6 is as defined for Formula I.
• the heterocyclic group is also independently optionally substituted on a ring nitrogen atom with alkyl, aralkyl, alkylcarbonyl, or on sulfur with lower alkyl.
• heterocyclic groups include, without limitation, epoxy, azetidinyl, aziridinyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, pyrrolidinonyl, piperidinyl, piperazinyl, imidazolidinyl, imidazopyridinyl, thiazolidinyl, dithianyl, trithianyl, dioxolanyl, oxazolidinyl, oxazolidinonyl, decahydroquinolinyl, piperidonyl, 4-piperidinonyl, quinuclidinyl, thiomorpholinyl, thiomorpholinyl 1,1 dioxide, morpholinyl, azepanyl, oxazepanyl, azabicyclohexanyls, azabicycloheptanyl, azabicyclooctanyls, azabicyclononanyls
• heteroaryl refers to groups having 5 to 14 ring atoms, preferably 5, 6, 9, or 10 ring atoms; having 6, 10, or 14 p electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to three heteroatoms per ring selected from the group consisting of N, O, and S.
• heteroaryl groups include acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, 6,7-dihydro-5H-pyrrolo[1,2- a]imidazole, furanyl, furazanyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl,
• Heteroaryl also refers to bicyclic ring systems having, in addition to carbon atoms, from one to three heteroatoms per ring selected from the group consisting of N, O, and S in which one ring system may be saturated or partially saturated.
• an effective amount of a compound is an amount that is sufficient to negatively modulate or inhibit the activity of KRas G12D. Such amount may be administered as a single dosage or may be administered according to a regimen, whereby it is effective.
• a "therapeutically effective amount" of a compound is an amount that is sufficient to ameliorate, or in some manner reduce a symptom or stop or reverse progression of a condition, or negatively modulate or inhibit the activity of KRas G12D. Such amount may be administered as a single dosage or may be administered according to a regimen, whereby it is effective.
• treatment means any manner in which the symptoms or pathology of a condition, disorder or disease are ameliorated or otherwise beneficially altered. Treatment also encompasses any pharmaceutical use of the compositions herein.
• amelioration of the symptoms of a particular disorder by administration of a particular pharmaceutical composition refers to any lessening, whether permanent or temporary, lasting or transient that can be attributed to or associated with administration of the composition.
• COMPOUNDS [0100]
• Y is a bond, O or NR 5 ;
• R 2 is hydrogen, -N(R 5 ) 2 , heterocyclyl, C1 – C6 alkyl, -L-he
• R 1 is hydrogen.
• R 1 is hydroxy.
• R 1 is -CO 2 R 5 .
• R 5 is hydrogen.
• R 5 is C1 – C3 alkyl.
• R 1 is -C(O) 2 N(R 5 ) 2 .
• each R 5 is hydrogen, each R 5 is an independently selected C1 – C3 alkyl, or one R 5 is hydrogen and the second R 5 is C1 – C3 alkyl.
• Y is a bond.
• R 2 is hydrogen, - N(R 5 )2, or heterocyclyl optionally substituted with one or more R 6 .
• R 2 is -N(R 5 ) 2 .
• each R 5 is hydrogen.
• each R 5 is an independently selected C1 – C3 alkyl.
• one R 5 is hydrogen and the second R 5 is C1 – C3 alkyl.
• Y is a bond and R 2 is - N(R 5 )2.
• R 2 is heterocyclyl.
• R 2 is heterocyclyl and the heterocyclyl is azetidinyl, pyrrolidinyl, tetrahydro-2H-thiopyran 1,1-dioxide or 1,6 ⁇ 2 - diazaspiro[3.3]heptanyl.
• Y is a bond and R 2 is heterocyclyl.
• the heterocyclyl is azetidinyl substituted with one R 6 .
• the heterocyclyl is azetidinyl substituted with one R 6 , wherein R 6 is hydroxy, hydroxyalkyl, or -N(R 5 )2. In certain embodiments, the heterocyclyl is azetidinyl substituted with two R 6 groups independently selected from -N(R 5 ) 2 and C1 – C3 alkyl. In certain embodiments, Y is a bond and the heterocyclyl is azetidinyl substituted with one R 6 , wherein R 6 is hydroxy, hydroxyalkyl, or -N(R 5 )2.
• Y is a bond and the heterocyclyl is azetidinyl substituted with two R 6 groups independently selected from -N(R 5 ) 2 and C1 – C3 alkyl. [0124] In one embodiment, Y is O.
• Y is O and R 2 is C1 – C6 alkyl. In certain embodiments, the C1 – C6 alkyl is methyl, ethyl, isopropyl or isobutyl. [0127] In one embodiment of the compounds of Formula (I), Y is O and R 2 is -L-heterocyclyl optionally substituted with one or more R 6 . [0128] In one embodiment, Y is O and R 2 is heterocyclyl wherein the heterocyclyl is tetrahydropyranyl optionally substituted with two halogens. In certain embodiment, the two halogens are both fluoro.
• Y is O and R 2 is -L-heterocyclyl wherein L is methylene and the heterocyclyl is hexahydro-1H-pyrrolizinyl, hexahydro-3H-pyrrolizin-3-one, hexahydro-1H- pyrrolo[2,1-c][1,4]oxazinyl, octahydroindolizinyl, hexahydropyrrolizine 4(1H)-oxide, azetidinyl, pyrrolidinyl, pyrrolidin-2-one, oxetanyl, piperidinyl, 1-azabicyclo[2.2.1]heptanyl, morpholinyl, oxa-5-azabicyclo[2.2.1]heptan-5-yl, thiopyranyl, 6-oxa-2 ⁇ 2 -azaspiro[3.4]octanyl, 7-oxa-2 ⁇
• Y is O and R 2 is -L-heterocyclyl wherein L is methylene and the heterocyclyl is hexahydro-1H-pyrrolizinyl.
• Y is O and R 2 is -L-heterocyclyl wherein L is methylene and the heterocyclyl is hexahydro-1H-pyrrolizinyl is optionally substituted with one R 6 , wherein R 6 is halogen, hydroxy, hydroxyalkyl, C1 - C3 haloalkyl, C1 – C3 alkyl, C1-C3 alkoxy, phenyl, tert- butyldimethylsilyloxyCH 2 - or pyrazolyl, wherein the pyrazolyl is optionally substituted with C1- C3 alkyl.
• the C1 - C3 haloalkyl is chloromethyl.
• the pyrazolyl is substituted with C1 – C3 alkyl.
• the hexahydro-1H- pyrrolizinyl is substituted with two R 6 groups, wherein each R 6 is an independently selected C1 – C3 alkyl.
• the heterocyclyl is hexahydro-1H-pyrrolizinyl which is unsubstituted.
• Y is O and R 2 is -L-heterocyclyl wherein L is methylene and the heterocyclyl is azetidinyl substituted with one R 6 , wherein R 6 is C1 – C3 alkyl.
• Y is O and R 2 is -L-heterocyclyl wherein L is methylene and the heterocyclyl is pyrrolidinyl substituted with one R 6 , wherein R 6 is C1 - C3 hydroxyalkyl, C1 - C3 haloalkyl, C1 – C3 alkyl, C1 - C3 alkoxy, C1-C3 aralkyl, or -Q-phenyl, wherein Q is O, and - NHC(O)phenyl.
• the phenyl group of the -Q-phenyl is substituted with SO 2 F.
• the phenyl group of the -NHC(O)phenyl is substituted with SO 2 F.
• the C1-C3 aralkyl is benzyl.
• Y is O and R 2 is -L-heterocyclyl wherein L is methylene and the pyrrolidinyl is substituted with two R 6 groups, wherein one R 6 is C1 – C3 alkyl and the other R 6 is C1 - C3 alkoxy or halogen.
• Y is O and R 2 is -L-heterocyclyl wherein L is methylene and the heterocyclyl is pyrrolidin-2-one substituted with one R 6 , wherein R 6 is C1 – C3 alkyl.
• Y is O and R 2 is -L-heterocyclyl wherein L is methylene and the heterocyclyl is piperidinyl substituted with one R 6 , wherein R 6 is acetyl, (C1-C3 alkoxy)C1-C3 alkoxy, or -C(O)CH 2 Cl.
• Y is O and R 2 is -L-heterocyclyl wherein L is methylene and the heterocyclyl is (2S)-1-azabicyclo[2.2.1]heptan-2-yl.
• Y is O, R 2 is -L-heterocyclyl wherein L is ethylene or propylene and the heterocyclyl is morpholinyl or oxa-5-azabicyclo[2.2.1]heptan- 5-yl.
• Y is O and R 2 is -L-heteroaryl, wherein the heteroaryl portion is optionally substituted with one or more R 7 .
• L is ethylene and the heteroaryl is benzimidazolyl, optionally substituted with one or more R 7 .
• R 7 is C1 – C4 alkyl.
• Y is O and R 2 is -L-heteroaryl.
• Y is O and R 2 is -L-heteroaryl, wherein L is methylene or ethylene.
• Y is O and R 2 is -L-heteroaryl, wherein L is methylene or ethylene and the heteroaryl is pyridyl, pyrazolyl, imidazolyl, triazolyl, 4,5,6,7-tetrahydro-1H-indazolyl, benzimidazolyl, imidazo[1,2-a]pyridinyl, or pyrimidinyl.
• Y is O and R 2 is -L-heteroaryl, wherein the heteroaryl is pyridyl substituted with one R 7 .
• Y is O and R 2 is -L-heteroaryl, wherein the heteroaryl is pyridyl substituted with one R 7 wherein R 7 is halogen, C1 – C4 haloalkyl, C1 – C4 hydroxyalkyl, C1 – C4 alkyl, -N(R 5 )2, or C1 – C4 alkoxy.
• R 7 is halogen, C1 – C4 haloalkyl, C1 – C4 hydroxyalkyl, C1 – C4 alkyl, -N(R 5 )2, or C1 – C4 alkoxy.
• Y is O and R 2 is -L-heteroaryl, wherein L is methylene or ethylene and the heteroaryl is pyrazolyl substituted with one R 7 .
• Y is O and R 2 is -L-heteroaryl, wherein L is methylene or ethylene and the heteroaryl is pyrazolyl substituted with one R 7 wherein R 7 is halogen, C1 – C4 haloalkyl, C1 – C4 hydroxyalkyl, C1 – C4 alkyl, alkoxy or -N(R 5 ) 2 .
• R 7 is halogen, C1 – C4 haloalkyl, C1 – C4 hydroxyalkyl, C1 – C4 alkyl, alkoxy or -N(R 5 ) 2 .
• Y is O and R 2 is -L-heteroaryl, wherein L is methylene or ethylene and the heteroaryl is imidazolyl substituted with one R 7 .
• Y is O and R 2 is -L-heteroaryl, wherein L is methylene or ethylene and the heteroaryl is imidazolyl substituted with one R 7 wherein R 7 is C1 – C4 alkyl, C1 – C4 haloalkyl, or C1 – C4 hydroxyalkyl.
• R 7 is C1 – C4 alkyl, C1 – C4 haloalkyl, or C1 – C4 hydroxyalkyl.
• Y is O and R 2 is -L-heteroaryl, wherein L is methylene or ethylene and the heteroaryl is triazolyl substituted with one R 7 .
• Y is O and R 2 is - L-heteroaryl, wherein L is methylene or ethylene and the heteroaryl is triazolyl substituted with one R 7 , wherein R 7 is C1 – C4 alkyl.
• Y is O and R 2 is -L-aryl, wherein the aryl portion is optionally substituted with one or more R 7 .
• L is ethylene and the aryl is phenyl.
• the phenyl is substituted with one R 7 .
• the phenyl is substituted with one R 7 , wherein R 7 is halogen.
• Y is O and R 2 is -L-cycloalkyl, wherein the cycloalkyl portion is optionally substituted with one or more R 6 .
• L is methylene.
• the cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
• the cyclopropyl and cyclopentyl are each substituted with one R 6 . In certain embodiments, the cyclopropyl and cyclopentyl are each substituted with one R 6 , wherein R 6 is haloalkyl. In certain embodiments, the cyclobutyl and cyclohexyl are each substituted with two R 6 groups. In certain embodiments, the cyclobutyl and cyclohexyl are each substituted with two R 6 groups, wherein each R 6 group is halogen. [0148] In one embodiment of the compounds of Formula (I), Y is O, and R 2 is -L-N(R 5 ) 2 . In certain embodiments, L is ethylene.
• R 5 is C1 – C3 alkyl.
• L is ethylene or propylene.
• Y is O, and R 2 is -L-C(O)N(R 5 ) 2 .
• L is ethylene and each R 5 is C1 – C3 alkyl.
• Y is O, and R 2 is -L-C1-C6 haloalkyl.
• L is methylene.
• the haloalkyl is 1,1,3,3- tetrafluoropropanyl or trifluoromethyl.
• L is ethylene or propylene and the haloalkyl is trifluoromethyl.
• Y is O
• R 2 is -L-COR 5 .
• L is propylene and R 5 is hydrogen or C1 – C3 alkyl.
• L is propylene that is substituted with hydroxy, hydroxyalkyl or heteroaryl and R 5 is hydrogen or C1 – C3 alkyl.
• the heteroaryl is pyridyl.
• Y is O, and R 2 is -L- (CH2OR 5 )(CH2)nOR 5 .
• L is methylene, each R 5 is independently hydrogen or C1 – C3 alkyl, and n is one or two.
• Y is O, and R 2 is -L-NR 5 C(O)-aryl.
• L is methylene, R 5 is hydrogen.
• the aryl is phenyl.
• the phenyl is substituted with one R 6 , wherein R 6 is -SO2F.
• R 3 is aryl optionally substituted with one or more R 8 .
• the aryl is selected from the group consisting of phenyl, naphthyl, 1,2,3,4-tetrahydronaphthalenyl and 2,3-dihydro-1H-indenyl, wherein each is optionally substituted with one or more R 8 .
• the aryl is phenyl substituted with one or more R 8 groups.
• the aryl is phenyl substituted with one or more R 8 groups independently selected from halogen, C1 - C3 haloalkyl and -O-C1 - C3 haloalkyl. In certain embodiments the phenyl is substituted with two R 8 groups. In certain embodiments the phenyl is substituted with two R 8 groups, wherein the two R 8 groups are two independently selected C1 - C3 haloalkyl groups, or - O-C1 - C3 haloalkyl and halogen. [0157] In one embodiment, the aryl is 2,3-dihydro-1H-indenyl optionally substituted with one or more R 8 .
• the aryl is 2,3-dihydro-1H-indenyl optionally substituted with one R 8 .
• R 8 is C1 – C alkyl.
• the aryl is naphthyl substituted with one or more R 8 groups.
• the aryl is naphthyl substituted with one or more R 8 groups independently selected from halogen, cyano, hydroxy, C1 - C3 alkyl, -S-C1 - C3 alkyl, C2 – C4 alkenyl, C2 – C4 alkynyl, C2 – C4 hydroxyalkynyl, C1-C3 cyanoalkyl, triazolyl, C1-C3 haloalkyl and -O-C1-C3 haloalkyl.
• the aryl is naphthyl substituted with hydroxy.
• the aryl is naphthyl substituted with halogen.
• the halogen is chlorine, fluorine or bromine. In other embodiments, the halogen is chlorine.
• the aryl is naphthyl substituted with C1 - C3 alkyl, wherein the C1 - C3 alkyl is methyl or ethyl.
• the aryl is naphthyl substituted with C2 – C4 alkenyl. In certain embodiments, the C2 – C4 alkenyl is prop-2-enyl.
• the aryl is naphthyl substituted with C2 – C4 alkynyl.
• the C2 – C4 alkynyl is ethyne or prop-2-ynyl.
• the aryl is naphthyl substituted with one or two R 8 , wherein each R 8 is halogen, cyano, hydroxy, C1 - C3 alkyl, -S-C1 - C3 alkyl, C2 – C4 alkenyl, C2 – C4 alkynyl, C2 – C4 hydroxyalkynyl, C1 – C3 cyanoalkyl, or triazolyl.
• the aryl is naphthyl substituted with two R 8 groups independently selected from halogen, hydroxy, C1 - C3 alkyl and C2 – C4 alkynyl.
• R 3 is heteroaryl optionally substituted with one or more R 8 .
• the heteroaryl is isoquinolinyl, indazolyl, or benzo[d][1,3]dioxolyl optionally substituted with one or more R 8 .
• the heteroaryl is indazolyl optionally substituted with one or more R 8 .
• the heteroaryl is indazolyl optionally substituted with C1-C3 alkyl.
• the heteroaryl is isoquinolinyl optionally substituted with one or more R 8 . In other embodiments, the heteroaryl is isoquinolinyl optionally substituted with halogen or C2-C4 alkynyl. In certain embodiments, the heteroaryl is benzo[d][1,3]dioxolyl optionally substituted with two R 8 groups. In certain embodiments, the heteroaryl is benzo[d][1,3]dioxolyl optionally substituted with two R 8 groups, wherein each R 8 group is an independently selected halogen. In one embodiment, the two halogens are gem-difluoro substitutions. [0165] In one embodiment of the compounds of Formula (I), R 4 is hydrogen.
• R 4 is halogen. In one embodiment, R 4 is fluorine. In one embodiment, R 4 is chlorine. [0167] In one embodiment of the compounds of Formula (I), R 4 is C1 – C3 alkyl. In one embodiment, R 4 is methyl.
• Nonlimiting examples of compounds of Formula (I) are selected from the group consisting of: , , , ,
• the compounds of Formula (I) include bis-hydrochloride, tris-hydrochloride, trifluoroacetic acid, bis-trifluoroacetic acid, and tris-trifluoracetic acid salts of the above compounds.
• the compounds of Formula (I) or pharmaceutically acceptable salt thereof may be formulated into pharmaceutical compositions.
• PHARMACEUTICAL COMPOSITIONS [0169]
• the invention provides pharmaceutical compositions comprising a KRas G12D inhibitor according to the invention and a pharmaceutically acceptable carrier, excipient, or diluent.
• Compounds of the invention may be formulated by any method well known in the art and may be prepared for administration by any route, including, without limitation, parenteral, intraperitoneal, intradermal, intracardiac, intraventricular, intracranial, intracerebrospinal, intrasynovial, intrathecal administration, intramuscular injection, intravitreous injection, intravenous injection, intra-arterial injection, oral, buccal, sublingual, transdermal, topical, intranasal, intratracheal, intrarectal, subcutaneous, and topical administration.
• compounds of the invention are administered intravenously in a hospital setting.
• administration may be by the oral route.
• the provided pharmaceutical compositions may be administered to a subject in need of treatment by injection systemically, such as by intravenous injection; or by injection or application to the relevant site, such as by direct injection via syringe, or direct application to the site when the site is exposed in surgery; or by topical administration.
• Parenteral administration can be by bolus injection or continuous infusion.
• Pharmaceutical compositions for injection may be presented in unit dosage form, e.g., in ampoules or in multi- dose containers, with an added preservative.
• the provided pharmaceutical compositions can also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection.
• the formulations may be modified with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
• suitable polymeric or hydrophobic materials for example as an emulsion in an acceptable oil
• ion exchange resins for example, as an emulsion in an acceptable oil
• sparingly soluble derivatives for example, as a sparingly soluble salt.
• the pharmaceutical compositions may, if desired, be presented in a vial, pack or a medical device, including but not limited to a dispenser device which may contain one or more unit dosage forms containing the active ingredient.
• the dispenser device can comprise a syringe having a single dose of the liquid formulation ready for injection.
• the syringe can be accompanied by instructions for administration.
• the characteristics of the carrier will depend on the route of administration.
• compositions according to the invention may contain, in addition to the inhibitor, diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art.
• diluents such as a cell, cell culture, tissue, or organism
• solubilizers such as a cell, cell culture, tissue, or organism
• the preparation of pharmaceutically acceptable formulations is described in, e.g., Remington's Pharmaceutical Sciences, 18th Edition, ed. A. Gennaro, Mack Publishing Co., Easton, Pa., 1990.
• the term pharmaceutically acceptable salt refers to salts that retain the desired biological activity of the above-identified compounds and exhibit minimal or no undesired toxicological effects.
• examples of such salts include, but are not limited to acid addition salts formed with inorganic acids (for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid.
• inorganic acids for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like
• organic acids such as acetic acid, oxalic acid, tartaric acid
• the compounds can also be administered as pharmaceutically acceptable quaternary salts known by those skilled in the art, which specifically include the quaternary ammonium salt of the formula -NR+Z-, wherein R is hydrogen, alkyl, or benzyl, and Z is a counterion, including chloride, bromide, iodide, -O-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (such as benzoate, succinate, acetate, glycolate, maleate, malate, citrate, tartrate, ascorbate, benzoate, cinnamoate, mandeloate, benzyloate, and diphenylacetate).
• R is hydrogen, alkyl, or benzyl
• Z is a counterion, including chloride, bromide, iodide, -O-alkyl, toluenesulfonate, methylsulf
• the active compound is included in the pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to a patient a therapeutically effective amount without causing serious toxic effects in the patient treated.
• a dose of the active compound for all of the above-mentioned conditions is in the range from about 0.01 to 300 mg/kg, for example 0.1 to 100 mg/kg per day, and as a further example 0.5 to about 25 mg per kilogram body weight of the recipient per day.
• a typical topical dosage will range from 0.01-3% wt/wt in a suitable carrier.
• the effective dosage range of the pharmaceutically acceptable derivatives can be calculated based on the weight of the parent compound to be delivered.
• the effective dosage can be estimated as above using the weight of the derivative, or by other means known to those skilled in the art.
• the pharmaceutical compositions comprising compounds of the present invention may be used in the methods of use described herein.
• METHODS OF USE [0177]
• the invention provides for methods for inhibiting KRas G12D activity in a cell, comprising contacting the cell in which inhibition of KRas G12D activity is desired with an effective amount of a compound of Formula (I), pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing the compound or pharmaceutically acceptable salt thereof.
• the contacting is in vitro.
• the contacting is in vivo.
• contacting refers to the bringing together of indicated moieties in an in vitro system or an in vivo system.
• "contacting" a KRas G12D with a compound provided herein includes the administration of a compound provided herein to an individual or patient, such as a human, having KRas G12D, as well as, for example, introducing a compound provided herein into a sample containing a cellular or purified preparation containing the KRas G12D.
• a cell in which inhibition of KRas G12D activity is desired is contacted with an effective amount of a compound of Formula (I) or pharmaceutically acceptable salt thereof to negatively modulate the activity of KRas G12D.
• a compound of Formula (I) or pharmaceutically acceptable salt thereof to negatively modulate the activity of KRas G12D.
• the methods described herein are designed to inhibit undesired cellular proliferation resulting from enhanced KRas G12D activity within the cell.
• the cells may be contacted in a single dose or multiple doses in accordance with a particular treatment regimen to effect the desired negative modulation of KRas G12D.
• the ability of compounds to bind KRas G12D may be monitored in vitro using well known methods, including those described in Examples A and B below.
• the inhibitory activity of exemplary compounds in cells may be monitored, for example, by measuring the inhibition of KRas G12D activity of the amount of phosphorylated ERK, for example using the method described in Example C below.
• methods of treating cancer in a patient in need thereof comprising administering to said patient a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound or pharmaceutically acceptable salt thereof are provided.
• compositions and methods provided herein may be used for the treatment of a KRas G12D-associated cancer in a patient in need thereof, comprising administering to said patient a therapeutically effective amount of a compound of Formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound or pharmaceutically acceptable salt thereof are provided.
• the KRas G12D-associated cancer is lung cancer.
• the compositions and methods provided herein may be used for the treatment of a wide variety of cancers including tumors such as lung, prostate, breast, brain, skin, cervical carcinomas, testicular carcinomas, etc.
• cancers that may be treated by the compositions and methods of the invention include, but are not limited to tumor types such as astrocytic, breast, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate and thyroid carcinomas and sarcomas.
• tumor types such as astrocytic, breast, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate and thyroid carcinomas and sarcomas.
• these compounds can be used to treat: Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinom
• the cancer is non-small cell lung cancer, small cell lung cancer, colorectal cancer, rectal cancer or pancreatic cancer. In certain embodiments, the cancer is non-small cell lung cancer.
• concentration and route of administration to the patient will vary depending on the cancer to be treated.
• the compounds, pharmaceutically acceptable salts thereof and pharmaceutical compositions comprising such compounds and salts also may be co-administered with other anti-neoplastic compounds, e.g., chemotherapy, or used in combination with other treatments, such as radiation or surgical intervention, either as an adjuvant prior to surgery or post-operatively.
• a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof as defined herein for use in therapy is also provided herein.
• a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof as defined herein for use in the treatment of cancer is also provided herein.
• a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for use in the inhibition of KRas G12D is also provided herein.
• a compound of Formula (I) or a pharmaceutically acceptable salt thereof as defined herein in the manufacture of a medicament for the treatment of cancer.
• a use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, as defined herein in the manufacture of a medicament for the inhibition of activity of KRas G12D is also provided herein.
• a compound of Formula (I) or a pharmaceutically acceptable salt thereof is also provided herein, in the manufacture of a medicament for the treatment of a KRas G12D-associated disease or disorder.
• a KRas G12D mutation e.g., a KRas G12D-associated cancer
• a regulatory agency-approved e.g., FDA- approved, assay or kit
• step B compound (2) is subjected to phosgene and then reacts with ammonia in a solvent such as dichloromethane and in the presence of a base such as N-ethyl-N-isopropylpropan-2-amine to form urea (3).
• step C the cyclization of compound (3) in the presence of a base such as cesium carbonate in a solvent such as toluene and at elevated temperature gives compound (4).
• step D dichloroazaquinazoline (5) is prepared from compound (4) with phosphoryl trichloride and N- ethyl-N-isopropylpropan-2-amine.
• step E compound (5) undergoes a S N Ar reaction with optionally substituted mono-Boc protected diazabicyclo[3.2.1]octane in a solvent such as dimethylformamide and in the presence of a base such as N-ethyl-N-isopropylpropan-2-amine to give compound (6).
• step F the substituent –Y-R 2 is introduced by substitution of the chlorine with a nucleophile having the formula H–Y-R 2 in a polar solvent such as dioxane in the presence of a base such as cesium carbonate to provide compound (7).
• step G the Boc group of compound (7) is removed using conditions known in the art, for example with cold 4 N HCl in a solvent such as dioxane, to provide compound (I).
• the species R 2 and/or R 3 will also contain protecting group(s), which can be removed before or after step G in the synthetic sequence.
• Compounds (1), (2), (3), (4), (5) (6) and (7) as shown and described above for Scheme I are useful as intermediates for preparing compounds of Formula (I) and are provided as further aspects of the invention.
• SCHEME II [0199] Compounds of Formula (I) wherein all of the substituents are as defined for Formula I, with the exception that –Y-R 2 is other than hydrogen, can be prepared according to Scheme II.
• step A the 4-chlorine of nicotinate derivative (8) is substituted with 2,4- dimethoxybenzylamine in a polar solvent such as dioxane and in the presence of a base such as N-ethyl-N-isopropylpropan-2-amine to give compound (9).
• step B compound (9) is coupled with an aryl boronic acid ester or aryl stannane under the Suzuki or Stille reaction conditions to give compound (10).
• step C the 2,4-dimethoxybenzyl group of compound (10) is removed with trifluoroacetic acid and in a solvent such as dichloromethane to give compound (11).
• step D compound (11) is treated with trichloroacetyl isocyanate in THF and then ammonia in methanol, and the cyclization is facilitated with heat to give pyridopyrimidinedione (12).
• step E dichloroazaquinazoline (13) is prepared from compound (12) with phosphoryl trichloride and N-ethyl-N-isopropylpropan-2-amine.
• step F compound (13) undergoes a S N Ar reaction with optionally substituted mono-Boc protected diazabicyclo[3.2.1]octane in a solvent such as N,N- dimethylacetamide and in the presence of a base such as N-ethyl-N-isopropylpropan-2-amine to give compound (14).
• step G the substituent –Y-R 2 is introduced by substitution of the chlorine with a nucleophile having the formula H–Y-R 2 in a polar solvent such as dioxane in the presence of a base such as cesium carbonate to provide compound (15).
• step H the Boc group of compound (15) is removed using conditions known in the art, for example with trifluoroacetic acid in a solvent such as dichloromethane, to provide compound (I).
• the species R 2 and/or R 3 will also contain protecting group(s), which can be removed before or after step H in the synthetic sequence.
• Compounds (8), (9), (10), (11), (12), (13), (14) and (15) as shown and described above for Scheme II are useful as intermediates for preparing compounds of Formula (I) and are provided as further aspects of the invention.
• step C trichloroazaquinazoline (18) is prepared from compound (17) with phosphoryl trichloride and N-ethyl-N-isopropylpropan-2- amine.
• step D compound (18) undergoes a SNAr reaction with optionally substituted mono- Boc protected diazabicyclo[3.2.1]octane to give compound (19) in a solvent such as N,N- dimethylacetamide and in the presence of a base such as N-ethyl-N-isopropylpropan-2-amine.
• step E the substituent –Y-R 2 is introduced by substitution of 2-chlorine of compound (19) with a nucleophile having the formula H–Y-R 2 in a polar solvent such as dioxane and in the presence of a base such as cesium carbonate to provide compound (20).
• step F compound (20) is coupled with an aryl boronic acid ester or aryl stannane under the Suzuki or Stille reaction conditions to give compound (15).
• step G the Boc group of compound (15) is removed using conditions known in the art, for example with trifluoroacetic acid in a solvent such as dichloromethane, to provide compound (I).
• the species R 2 and/or R 3 will also contain protecting group(s), which can be removed before or after step G in the synthetic sequence.
• Compounds (16), (17), (18), (19), and (20) as shown and described above for Scheme III are useful as intermediates for preparing compounds of Formula (I) and are provided as further aspects of the invention.
• SCHEME IV [0203] Compounds of Formula (I) wherein all of the substituents are as defined for Formula I, with the exception that –Y-R 2 is other than hydrogen, can be prepared according to Scheme IV.
• step A 4-chlorine of trichloroazaquinazoline (18) is substituted with a benzyl alcohol in a polar solvent such as dioxane and in the presence of a base such as N-ethyl-N-isopropylpropan-2-amine to provide compound (21).
• step B the substituent –Y-R 2 is introduced by substitution of 2- chlorine of compound (21) with a nucleophile having the formula H–Y-R 2 in a polar solvent such as dioxane and in the presence of a base such as cesium carbonate to provide compound (20).
• step C compound (22) is coupled with an aryl boronic acid ester or aryl stannane under the Suzuki or Stille reaction conditions to give compound (23).
• step D the benzyl group of compound (23) is removed under the palladium-catalyzed hydrogenation condition in a solvent such as ethyl acetate to give compound (24).
• step E compound (24) is coupled with optionally substituted mono-Boc protected diazabicyclo[3.2.1]octane to provide compound (15).
• step F the Boc group of compound (15) is removed using conditions known in the art, for example with trifluoroacetic acid in a solvent such as dichloromethane, to provide compound (I).
• the species R 2 and/or R 3 will also contain protecting group(s), which can be removed before or after step G in the synthetic sequence.
• step B the chlorination of compound 26 with phosphoryl trichloride and N-ethyl-N-isopropylpropan- 2-amine provides dichloroazaquinazoline (27).
• step C compound (27) undergoes a SNAr reaction with optionally substituted mono-Boc protected diazabicyclo[3.2.1]octane in a solvent such as N,N-dimethylacetamide and in the presence of a base such as N-ethyl-N-isopropylpropan- 2-amine to give compound (28).
• step D compound (28) is coupled with an aryl boronic acid ester or aryl stannane under the Suzuki or Stille reaction conditions to give compound (29).
• step E the Boc group of compound (29) is removed using conditions known in the art, for example with cold 4N HCl and in a solvent such as dioxane, to provide compound (30).
• the species R 3 will also contain a protecting group, which can be removed before or after step G in the synthetic sequence.
• Compounds (25), (26), (27), (28), and (29) as shown and described above for Scheme V are useful as intermediates for preparing compounds of Formula (I) and are provided as further aspects of the invention.
• SCHEME VI [0207] Compounds of Formula (I) wherein all substituents are as defined for Formula I, with the exception that –Y-R 2 is other than hydrogen, can be prepared according to Scheme VI.
• step A compound (14) undergoes a Sonogashira coupling reaction in a polar solvent such as acetonitrile to provide compound (15).
• step B the Boc group of compound (15) is removed using conditions known in the art, for example with trifluoroacetic acid in a solvent such as dichloromethane, to provide compound (I).
• the species R 2 and/or R 3 will also contain protecting/masking group(s), which can be removed before or after step B in the synthetic sequence.
• the compounds of the present invention may have one or more chiral center and may be synthesized as stereoisomeric mixtures, isomers of identical constitution that differ in the arrangement of their atoms in space.
• the compounds may be used as mixtures or the individual components/isomers may be separated using commercially available reagents and conventional methods for isolation of stereoisomers and enantiomers well-known to those skilled in the art, e.g., using CHIRALPAK® (Sigma-Aldrich) or CHIRALCEL® (Diacel Corp) chiral chromatographic HPLC columns according to the manufacturer’s instructions.
• CHIRALPAK® Sigma-Aldrich
• CHIRALCEL® Diacel Corp
• compounds of the present invention may be synthesized using optically pure, chiral reagents and intermediates to prepare individual isomers or enantiomers. Unless otherwise indicated, all chiral (enantiomeric and diastereomeric) and racemic forms are within the scope of the invention.
• Step B 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl pivalate.
• PdCl 2 (dppf) (0.287 g, 0.393 mmol) was added and the reaction heated to 95°C for 6 h and then stirred at room temperature for 16 h. The mixture was partitioned between water (100 mL) and EtOAc (50 mL) and the aqueous layer was extracted with EtOAc (2 x 30 mL). The combined organic phases were washed with brine (30 mL), dried over Na2SO4, filtered and concentrated in vacuo.
• Step B 2-chloro-3-fluoro-pyridin-4-amine.
• HCl/dioxane 4 M, 796 mL, 3.95 eq. The mixture was stirred at 25 °C for 2 hours.
• Step C 2-chloro-3-fluoro-5-iodo-pyridin-4-amine.
• 2-chloro-3-fluoro- pyridin-4-amine 107 g, 730 mmol, 1.0 eq
• NIS 197 g, 876 mmol, 1.2 eq
• MeCN MeCN
• p-toluene sulfonic acid monohydrate 6.94 g, 36.5 mmol, 0.05 eq.
• the mixture was stirred at 70 °C for 16 hours. Upon completion, the mixture was diluted with water (300 mL) and ethyl acetate (2000 mL).
• ethyl-6-chloro-5-fluoro-4-[(2,2,2-trichloroacetyl)carbamoylamino]pyridine-3- carboxylate To a solution of ethyl 4-amino-6-chloro-5-fluoro-pyridine-3-carboxylate (20.3 g, 73.2 mmol, 1.0 eq) in THF (60 mL) was added 2,2,2-trichloroacetyl isocyanate (20.7 g, 110 mmol, 13.0 mL, 1.5 eq) at 25 °C. The mixture was stirred at 25 °C for 10 min. Upon completion, the mixture was concentrated under vacuum.
• Step B tert-butyl-3-(2,7-dichloro-8-fluoro-pyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate.
• Step B 2,7-dichloro-8-fluoro-4-(2,2,2-trifluoroethoxy)pyrido[4,3-d]pyrimidine.
• 2,2,2-trifluoroethanol 11.1 g, 111 mmol, 8.01 mL, 1.20 eq
• t-BuONa 26.7 g, 278 mmol, 3.00 eq
• the mixture was first stirred at 10 °C for 0.5 hour. Then the above mixture was added to 2,4,7-trichloro-8-fluoro-pyrido[4,3-d]pyrimidine (23.4 g, 92.7 mmol, 1.00 eq) in toluene (200 mL) at -10 °C. After addition, the mixture was stirred at -10 °C ⁇ 25 °C for 16 hours.
• the mixture was stirred at 0 - 25 °C for 2 hours. After completion, the mixture was filtered and washed with ethyl acetate (100 mL). The filtrate was quenched by saturated NH4Cl aqueous solution (300 mL), and the organic layer was separated and dried over anhydrous Na 2 SO 4 . The mixture was filtered, and the filtrate was concentrated under reduced pressure at 40 °C to dryness. The crude product was triturated with CH3CN (20 mL) at 25 °C for 15 minutes and filtered, the filter cake was dried in vacuum at 40 °C affording the title compound (18.2 g, 64.6% yield). Light yellow solid.
• Step A methyl 4-(tert-butoxycarbonylamino)-6-chloro-5-fluoro-pyridine-3-carboxylate.
• 4-((tert-butoxycarbonyl)amino)-6-chloro-5-fluoronicotinic acid (14.3 g, 49.2 mmol, 1 eq) in MeOH (70 mL) and toluene (210 mL) was added TMSCHN 2 (2 M in hexane, 44.3 mL, 1.8 eq) slowly.
• Step B methyl 4-amino-6-chloro-5-fluoro-pyridine-3-carboxylate.
• Step C methyl 4-amino-6-(8-chloro-1-naphthyl)-5-fluoro-pyridine-3-carboxylate.
• Step D methyl 6-(8-chloro-1-naphthyl)-5-fluoro-4-[(2,2,2- trichloroacetyl)carbamoylamino] pyridine-3-carboxylate.
• Step A utyl 3-[2-chloro-7-(8-chloro-1-naphthyl)-8-fluoro-pyrido[4,3-d]pyrimidin-4- yl]-3,8- diazabicyclo[3.2.1]octane-8-carboxylate.
• Step A tert-butyl N-(2-chloro-3-fluoro-4-pyridyl)carbamate.
• Step E 4-amino-6-chloro-5-fluoro-pyridine-3-carboxamide.
• H 2 SO 4 146 g, 1.46 mol, 79.3 mL, 98% purity, 5.0 eq
• 4-amino-6-chloro-5-fluoro-pyridine-3-carbonitrile 50 g, 291 mmol, 1.0 eq
• the reaction mixture was heated to 60 °C for 1h.
• the reaction mixture was poured into ice water (1 L) with stirring. A yellow solid was precipitated.
• Step B 4,7-dichloro-8-fluoropyrido[4,3-d]pyrimidine.
• 7-chloro-8- fluoropyrido[4,3-d]pyrimidin-4-ol 2.2 g, 11.0 mmol, 1.0 eq
• N-ethyl-N-isopropylpropan-2- amine 2.85 g, 22.0 mmol, 3.84 mL, 2 eq
• POCl3 82.5 g, 538 mmol, 50 mL, 48.8 eq.
• the mixture was stirred at 110 °C for 3 h.
• Step C ( utyl 3-(7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo [3.2.1]octane-8-carboxylate.
• Step C.1-bromo-8-chloronaphthalene To a solution of 8-chloronaphthalen-1-amine (57 g, 320 mmol, 1 eq) and TsOH•H 2 O (219 g, 1.16 mol, 3.6 eq) in MeCN (1000 mL) was added a solution of NaNO2 (39.8 g, 577 mmol, 1.8 eq) and CuBr (138 g, 963 mmol, 29.3 mL, 3 eq) in H2O (120 mL) at - 5 °C, then the reaction mixture was stirred at 25 °C for 12 hours.
• Step A Benzyl carbonazidate. Benzyl carbonochloridate (100 mg, 586 ⁇ mol, 83.3 ⁇ L, 1.0 equivalent) was added to a well-stirred suspension of NaN3 (45.7 mg, 703 ⁇ mol, 1.2 equivalent) in acetone (10 mL) at 10 °C. The mixture was stirred at 10 °C for 1 hour.
• Step B tert-butyl 4-(4-fluorophenyl)-3-oxobutanoate.
• Step C 4-(4-fluorophenyl)-3-oxobutanoic acid.
• Step B ethyl 2,5-dioxotetrahydro-1H-pyrrolizine-7a(5H)-carboxylate.
• Step C ethyl 2-hydroxy-5-oxotetrahydro-1H-pyrrolizine-7a(5H)-carboxylate.
• EtOH ethyl 2,5-dioxotetrahydro-1H-pyrrolizine-7a(5H)-carboxylate (257 g, 1.22 mol, 1.00 eq) in EtOH (1300 mL) was slowly added NaBH4 (13.8 g, 365 mmol, 0.30 eq) at 0 °C under N2. The mixture was stirred at 0 °C for 10 min.
• [3-(methoxymethoxy)-8-(2-triisopropylsilylethynyl)-1- naphthyl]trifluoromethanesulfonate To a mixture of 3-(methoxymethoxy)-8-(2- triisopropylsilylethynyl)naphthalen-1-ol (200 g, 520.04 mmol, 1 eq) and DIEA (202 g, 1.56 mol, 272 mL, 3 eq) in dichloromethane (2000 mL) was added Tf2O (220 g, 780 mmol, 129 mL, 1.5 eq) at -40 °C.
• Step C 8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl pivalate.
• MeOH 20 mL
• Pd/C 200 mg, 10% purity
• Step D.8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-ol To the solution of 8-ethyl- 7-fluoro-3-(methoxymethoxy)naphthalen-1-yl pivalate (1.00 g, 2.99 mmol, 1.0 eq) in MeOH (15 mL) was added KOH (504 mg, 8.98 mmol, 3.0 eq), and the mixture was stirred at 20 °C for 0.5 hour.
• reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (3 ⁇ 10 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by reversed phase flash chromatography (C18, 0.1% FA in water, 0-90% ACN) affording the title compound (420 mg, two steps yield: 32%). Yellow solid.
• the mixture was stirred at 25 °C for 16 hours.
• the reaction mixture was quenched with saturated NaHCO 3 aqueous solution (100 mL) below 5 °C and diluted with H 2 O (300 mL).
• the organic layer was separated and H2O (100 mL) was added.
• the pH of the mixture was adjusted to 3 ⁇ 4 with 2N HCl below 10 °C.
• the organic layer was separated, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue.
• Step A Ethyl 4-amino-6-(3-(pivaloyloxy)naphthalen-1-yl)nicotinate.
• ethyl 4-amino-6-chloronicotinate (0.850 g, 4.24 mmol) in dioxane was added potassium carbonate (2.00 M solution, 10.6 ml, 21.2 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)naphthalen-2-yl pivalate (2.25 g, 6.36 mmol) and the reaction was sparged with nitrogen for 15 min.
• Step B Ethyl 6-(3-(pivaloyloxy)naphthalen-1-yl)-4-ureidonicotinate.
• Step A Ethyl 4,6-dichloro-5-fluoronicotinate.
• a solution of 4,6-dichloro-5-fluoro-3- pyridinecarboxylic acid (10.0 g, 47.6 mmol) in ethanol (238 ml, 47.6 mmol) was heated at 80°C and thionyl chloride (6.95 ml, 95.2 mmol) was added dropwise through the condenser. The mixture was stirred at 65°C overnight. The reaction was concentrated in vacuo and the residue was partitioned between EtOAc/water.
• Step C Ethyl 4-( imethoxybenzyl)amino)-5-fluoro-6-(3-(pivaloyloxy)naphthalen-1- yl)nicotinate.
• Ethyl 4-amino-5-fluoro-6-(3-(pivaloyloxy)naphthalen-1-yl)nicotinate A mixture of ethyl 4-((2,4-dimethoxybenzyl)amino)-5-fluoro-6-(3-(pivaloyloxy)naphthalen-1-yl)nicotinate (810 mg, 1.44 mmol) in CH2Cl2 (14.4 ml) was treated with TFA (1.11 ml, 14.4 mmol) at 0°C. After stirring at RT for 1 h, the reaction mixture was diluted with a solution of saturated NaHCO 3 , extracted with EtOAc.
• Step I 4-(4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-(((S)-1- methylpyrrolidin-2-yl)methoxy)pyrido[4,3-d]pyrimidin-7-yl)naphthalen-2-ol bis(2,2,2- trifluoroacetate).
• Step A Ethyl 4,6-dichloro-5-fluoronicotinate: A solution of 4,6-dichloro-5-fluoro-3- pyridinecarboxylic acid, (50 g, 238 mmol) in ethanol (1191 mL) was heated at 80 °C and thionyl chloride (34.8 mL, 476 mmol) was added dropwise. The mixture was stirred at 65 °C then cooled and concentrated in vacuo. The residue was partitioned between EtOAc and water and the organic phase was washed with sat.
• Ethyl 6-chloro-4-((2,4-dimethoxybenzyl)amino)-5-fluoronicotinate To a mixture of ethyl 4,6-dichloro-5-fluoronicotinate (50.4 g, 212 mmol) and N-ethyl-N-isopropylpropan-2- amine (92.4 mL, 529 mmol) in dioxane (605 mL) was added 2,4-dimethoxybenzylamine (35.0 mL, 233 mmol) and the mixture stirred at 50 °C for 18 h. The mixture was partitioned between EtOAc and water and the aqueous layer extracted with EtOAc (2x).
• Ethyl 4-amino-6-chloro-5-fluoronicotinate A solution of ethyl 6-chloro-4-((2,4- dimethoxybenzyl)amino)-5-fluoronicotinate (1.06 g, 2.87 mmol) in CH2Cl2 (19.2 mL) at 0 °C was treated dropwise with TFA (4.43 mL, 57.5 mmol). The mixture was stirred for 45 min, then diluted with CH2Cl2 (30.0 mL), and treated with 1M K3PO4 (30.0 mL). The mixture was filtered through GF paper and the filtrate layers were separated.
• Step D.7-Chloro-8-fluoropyrido[4,3-d]pyrimidine-2,4(1H,3H)-dione To a suspension of ethyl 4-amino-6-chloro-5-fluoronicotinate (628 mg, 2.87 mmol) in THF (6 mL) cooled to 0 °C was added trichloroacetyl isocyanate (0.410 mL, 3.45 mmol). The mixture was stirred at rt for 30 min then concentrated in vacuo. The residue was suspended in MeOH (14.4 mL), cooled to 0 °C and treated with ammonia (7M in MeOH, 14.4 mL, 101 mmol).
• Step E.2,4,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidine A mixture of 7-chloro-8- fluoropyrido[4,3-d]pyrimidine-2,4-diol (663 mg, 3.08 mmol) in phosphorous oxychloride (15.4 mL, 3.08 mmol) was treated with N-ethyl-N-isopropylpropan-2-amine (1.54 mL, 3.08 mmol). The mixture was heated to 110 °C where it stirred for 18 h. The cooled mixture was concentrated in vacuo. The residue was dissolved in CH 2 Cl 2 and washed with water (3x) and sat. NaHCO 3 .
• Step G tert-butyl (1R,5S)-3-(7-chloro-8-fluoro-2-(2-(1-methyl-1H-imidazol-2- yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate: A mixture of tert-butyl (1R,5S)-3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (50.0 mg, 0.117 mmol) in dioxane (11.7 mL) was treated with 2-(1-methyl-1H-imidazol-2-yl)ethan-1-ol (29.5 mg, 0.233 mmol) and Cs2CO3 (114 mg, 0.350 mmol) and stirred at 70 °
• Step H tert-Butyl (1R,5S)-3-(7-chloro-8-fluoro-2-(2-(1-methyl-1H-imidazol-2- yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate: A mixture of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-ol (26.8 mg, 0.099 mmol), tert- butyl (1R,5S)-3-(7-chloro-8-fluoro-2-(2-(1-methyl-1H-imidazol-2-yl)ethoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8-di
• Step I 4-(4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-(2-(1-methyl-1H- imidazol-2-yl)ethoxy)pyrido[4,3-d]pyrimidin-7-yl)naphthalen-2-ol: To a solution of tert-butyl (1R,5S)-3-(8-fluoro-7-(3-hydroxynaphthalen-1-yl)-2-(2-(1-methyl-1H-imidazol-2- yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (23.7 mg, 0.038 mmol) in CH 2 Cl 2 (0.760 mL
• Step A 2-(2,3-dihydro-1H-inden-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane.
• 4-bromo-2,3-dihydro-1H-indene 500 mg, 2.54 mmol
• dioxane 12.7 ml
• potassium acetate 747 mg, 7.61 mmol
• 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2- dioxaborolane) (1.93 g, 7.61 mmol).
• the reaction was sparged with N2 for 15 minutes, followed by addition of PdCl2(dppf) (186 mg, 0.254 mmol) and the reaction was heated to 95°C for 18 hrs.
• Step A 1-(1-methylpyrrolidin-2-yl)ethan-1-ol.
• Step B The title compound was synthesized according to Example 3, Steps G-I substituting 1-(1-methylpyrrolidin-2-yl)ethan-1-ol in place of 2-(1-methyl-1H-imidazol-2-yl)ethan-1-ol (4.00 mg, 83%).
• LCMS MM-ES+APCI, Pos: m/z 529.3 (M+H).
• Step A Ethyl 4,6-dichloro-5-fluoronicotinate.
• a solution of 4,6-dichloro-5-fluoro-3- pyridinecarboxylic acid (10.0 g, 47.6 mmol) in ethanol (238 ml, 47.6 mmol) was heated at 80°C and thionyl chloride (6.95 ml, 95.2 mmol) was added dropwise through the condenser. The resulting mixture was stirred at 65°C overnight. The reaction mixture was concentrated, and the residue was partitioned between EtOAc/water.
• Step B Ethyl 6-chloro-4-((2,4-dimethoxybenzyl)amino)-5-fluoronicotinate.
• Step D Ethyl 4-amino-6-(8-chloronaphthalen-1-yl)-5-fluoronicotinate.
• ethyl 6-(8-chloronaphthalen-1-yl)-4-((2,4-dimethoxybenzyl)amino)-5-fluoronicotinate (2.51 g, 5.07 mmol) in DCM (35 mL) was added TFA (7.81 mL, 101 mmol). The mixture was stirred for 1.5 hours and then carefully basified with 1M K3PO4.
• Step H tert-butyl (1R,5S)-3-(7-(8-chloronaphthalen-1-yl)-8-fluoro-2-(((S)-1- isopropylpyrrolidin-2-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane- 8-carboxylate.
• Step I 4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-chloronaphthalen-1-yl)-8- fluoro-2-(((S)-1-isopropylpyrrolidin-2-yl)methoxy)pyrido[4,3-d]pyrimidine.
• Step A ethyl 6-(3-(benzyloxy)naphthalen-1-yl)-4-((2,4-dimethoxybenzyl)amino)-5- fluoronicotinate.
• 2-(3-(benzyloxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.75 g, 3.25 mmol)
• Pd(PPh 3 ) 4 313 mg, 0.271 mmol
• Step B ethyl 4-amino-6-(3-(benzyloxy)naphthalen-1-yl)-5-fluoronicotinate.
• ethyl 6-(3-(benzyloxy)naphthalen-1-yl)-4-((2,4-dimethoxybenzyl)amino)-5-fluoronicotinate (1.21 g, 2.14 mmol) in dry DCM (14.2 mL) was added TFA (3.29 mL, 42.7 mmol). The mixture was stirred for 1 h, then carefully basified with 1M K3PO4, and filtered through GF paper.
• Step F tert-butyl (1R,5S)-3-(7-(3-(benzyloxy)naphthalen-1-yl)-8-fluoro-2-(2-(1-methyl- 1H-benzo[d]imidazol-2-yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane- 8-carboxylate.
• Step A Tert-butyl (1S,4S,5S)-5-hydroxy-7-azabicyclo[2.2.1]hept-2-ene-7-carboxylate.
• tert-butyl -5-oxo-7-azabicyclo[2.2.1]hept-2-ene-7-carboxylate 500 mg, 2.39 mmol
• solid sodium borohydride 45 mg, 1.19 mmol
• the reaction was partitioned between water (20 mL) and EtOAc (30 mL), and the layers were separated.
• Step D Tert-butyl 3-(2,4-dimethoxybenzyl)-6-((triethylsilyl)oxy)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate and tert-butyl 3-(2,4-dimethoxybenzyl)-6-hydroxy-3,8- diazabicyclo[3.2.1]octane-8-carboxylate.
• Step E Tert-butyl 3-(2,4-dimethoxybenzyl)-6-((triethylsilyl)oxy)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate: To a stirred solution of tert-butyl 3-(2,4- dimethoxybenzyl)-6-((triethylsilyl)oxy)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (251 mg, 0.509 mmol) in MeOH (4 mL) was added 20% palladium hydroxide on carbon (200 mg) and the reaction mixture was degassed.
• Step F Tert-butyl 3-(8-fluoro-7-(3-hydroxynaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin- 2-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-6-((triethylsilyl)oxy)-3,8-diazabicyclo[3.2.1]octane- 8-carboxylate.
• Step G 3-(8-fluoro-7-(3-hydroxynaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2- yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-6-ol.
• tert-butyl (1S,4S,5S)-5-((tert-butyldimethylsilyl)oxy)-7-azabicyclo[2.2.1]hept-2-ene-7- carboxylate A mixture of tert-butyl (1S,4S,5S)-5-hydroxy-7-azabicyclo[2.2.1]hept-2-ene-7- carboxylate (4.1 g, 19 mmol), imidazole (2.0 g, 29 mmol), and tert-butylchlorodimethylsilane (3.5 g, 23 mmol) in N,N-dimethylformamide (8 mL) was stirred at 30 °C for 1 hour and then at r.t. overnight.
• Step E (1R,5R,6R)-3-(7-(8-chloronaphthalen-1-yl)-8-fluoro-2-(2-(5-fluoropyridin-2- yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-6-ol.
• Step B tert-butyl (1R,5S)-3-(7-(5-chloroisoquinolin-4-yl)-8-fluoro-2-((tetrahydro-1H- pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8- carboxylate: A mixture of tert-butyl (1R,5S)-3-(7-chloro-8-fluoro-2-((tetrahydro-1H-pyrrolizin- 7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (50 mg, 0.0
• PdCl 2 (dppf) (6.9 mg, 0.009 mmol) was added and the mixture was stirred in a sealed tube at 90°C for 16 hours.
• the cooled mixture was partitioned between EtOAc (15 mL) and water (20 mL), and then filtered through GF paper.
• the aqueous layer was extracted with EtOAc (2x10 mL) and the combined organic phases were washed with brine (20 mL), dried over Na 2 SO 4 , filtered and concentrated in vacuo.
• Step C 4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(5-chloroisoquinolin-4-yl)-8- fluoro-2-((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidine bis(2,2,2- trifluoroacetate): To solution of tert-butyl (1R,5S)-3-(7-(5-chloroisoquinolin-4-yl)-8-fluoro-2- ((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-car
• 1-(tert-butyl) 2-methyl (2S,4R)-4-methoxypyrrolidine-1,2-dicarboxylate A mixture of 1-(tert-butyl) 2-methyl (2S,4R)-4-hydroxypyrrolidine-1,2-dicarboxylate (500 mg, 2.04 mol), iodomethane (0.330 ml, 5.30 mmol) and silver oxide (520 mg, 2.24 mmol) in acetonitrile (2.1 ml) was stirred at room temperature overnight. Additional silver oxide (520 mg, 2.24 mmol) and iodomethane (0.330 ml, 5.30 mmol) were added and the reaction was stirred at room temperature overnight.
• Step A 2-(((4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-chloronaphthalen-1-yl)- 8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)propane-1,3-diol:
• Step A 2,4-dibromo-5-chloronaphthalen-1-amine: To a solution of 5-Chloronaphthalen- 1-amine (1000 mg, 5.63 mmol) in chloroform (30 ml) was added bromine (0.58 ml, 11.3 mmol) in chloroform (30 ml) dropwise. The mixture was heated at 50° C overnight. Additional bromine (0.58 ml, 11.3 mmol) in 30 ml of chloroform was added dropwise at room temperature and the mixture was warmed to 50° C for 4 more hours. The reaction was cooled to rt and concentrated in vacuo. Water was added to the residue and the aqueous layer was extracted three times with ethyl acetate.
• 1-bromo-8-chloro-3-(methoxymethoxy)naphthalene To a solution of 4-bromo- 5-chloronaphthalen-2-ol (203 mg, 0.788 mmol) in THF (3900 ⁇ L) at 0° C was added sodium hydride (47.3 mg, 1.18 mmol). The mixture was stirred at 0° C for 30 minutes followed by addition of chloromethyl methyl ether (77.8 ⁇ L, 1.02 mmol) and the mixture was warmed to room temperature over 2 hours. The reaction was concentrated in vacuo. The residue was partitioned between EtOAc and water and the layers were separated. The aqueous layer was extracted with additional ethyl acetate.
• tert-butyl (1R,5S)-3-(7-chloro-8-fluoro-2-(3-hydroxyazetidin-1-yl)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate To a mixture of tert-butyl (1R,5S)-3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8- carboxylate (100 mg, 0.233 mmol) in dioxane (4 ml) were added azetidin-3-ol (HCl salt, 95%; 27 mg, 0.234 mmol), and Cs2CO3 (228 mg, 0.700 mmol) at rt.
• Step B The title compound was synthesized according to Example 3, Step H substituting tert-butyl (1R,5S)-3-(7-chloro-8-fluoro-2-(3-hydroxyazetidin-1-yl)pyrido[4,3-d]pyrimidin-4-yl)- 3,8-diazabicyclo[3.2.1]octane-8-carboxylate in place of tert-butyl (1R,5S)-3-(7-chloro-8-fluoro- 2-(2-(1-methyl-1H-imidazol-2-yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate followed by Example 2, step I (1.16 mg, 0.0024 mmol, 70%).
• Step B The title compound was synthesized according to Example 3, Step G and H substituting (1R,2S,4R)-1-azabicyclo[2.2.1]heptan-2-yl)methanol in place of 2-(1-methyl-1H- imidazol-2-yl)ethan-1-ol followed by deprotection using Example 2, Step I (15.59 mg, 0.030 mmol, 22% yield).
• LCMS MM-ES+APCI, Pos
• Step A 8-bromo-1-chloronaphthalen-2-amine: To a mixture of 8-bromonaphthalen-2- amine (2.3 g, 10 mmol) in CCl4 (104 ml, 10 mmol) was added NCS (1.4 g, 10 mmol) and the resulting mixture was heated at 50 °C for 2h. The mixture was cooled to rt and stirred overnight. The slurry was filtered to isolate solid product. The solid was purified by column chromatography eluting with 0-20% hex/EtOAc to give product (1.9g, 72%).
• Step B 8-bromo-1-chloro-2-fluoronaphthalene: A slurry of nitrosonium tetrafluoroborate (378 mg, 3.23 mmol) in DCM (2.5 ml) was cooled in an ice bath. 8-bromo-1-chloronaphthalen- 2-amine (680 mg, 2.65 mmol) was added. The mixture was stirred for 1 hour and the solvent was removed. 1,2-dichlorobenzene (10 ml) was added and the reaction mixture was heated at 160 to 180° C for one hour.
• Step E 4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-chloro-7-fluoronaphthalen-1- yl)-8-fluoro-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[4,3-d]pyrimidine
• the title compound was synthesized according to the method of Example 2, step I (6.93 mg, 0.093 mmol, 86% yield).
• EXAMPLE 45 4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-chloro-7-fluoronaphthalen-1- yl)-8-fluoro-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[4,3-
• EXAMPLE 48 4-(4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-(2,2,2-trifluoroethoxy)pyrido[4,3- d]pyrimidin-7-yl)naphthalen-2-ol [0399] Synthesized according to Example 3, Step G and H substituting 2,2,2-Trifluoroethanol in place of 2-(1-methyl-1H-imidazol-2-yl)ethan-1-ol followed by deprotection according to the method of Example 2, Step I (42.15 mg, 0.0606 mmol, 33% yield). LCMS (MM-ES+APCI, Pos): m/z 500.2 [M+H]. EXAMPLE 49
• Step A 3-(7-(8-chloronaphthalen-1-yl)-8-fluoro-2-((tetrahydro-1H-pyrrolizin-7a(5H)- yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-6-ol : A mixture tert- butyl 6-((tert-butyldimethylsilyl)oxy)-3-(7-(8-chloronaphthalen-1-yl)-8-fluoro-2-((tetrahydro- 1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane- 8-carboxylate (27 mg, 0.034200 mmol) was placed in 7:3 DCM:TFA (1 m
• Step B 7a-(((4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-chloronaphthalen-1-yl)- 8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)hexahydropyrrolizine 4(1H)-oxide.
• the reaction was sparged with argon for 5 minutes and then palladium (II) acetate (2.02 mg, 0.009 mmol) was added.
• the reaction was heated at 110° C for 3 hours.
• the reaction was cooled to rt, water was added, and the aqueous layer was extracted 2X with ethyl acetate. The pooled organic layers were dried over magnesium sulfate, filtered, and concentrated.
• reaction solution was partitioned between sat. NaHCO3 and ethyl acetate and the layers were separated. The aqueous layer was extracted with additional ethyl acetate. The pooled organic layers were dried over magnesium sulfate, filtered, and concentrated.
• Step B tert-butyl (1R,5S)-3-(2-((3-(chloromethyl)tetrahydro-1H-pyrrolizin-7a(5H)- yl)methoxy)-7-(8-chloronaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate: A solution of tert-butyl (1R,5S)-3-(7-(8- chloronaphthalen-1-yl)-8-fluoro-2-((3-(hydroxymethyl)tetrahydro-1H-pyrrolizin-7a(5H)- yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo
• Step C 4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((3R,7aS)-3- (chloromethyl)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7-(8-chloronaphthalen-1-yl)-8- fluoropyrido[4,3-d]pyrimidine and 4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((3S,7aS)- 3-(chloromethyl)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7-(8-chloronaphthalen-1-yl)-8- fluoropyrido[4,3-d]pyrimidine : Synthesized according to Example 2, Step I to yield two isomeric products.
• EXAMPLE 70 4-(4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-(2-methoxyethoxy)pyrido[4,3- d]pyrimidin-7-yl)naphthalen-2-ol [0428] Synthesized similarly to Example 3 Steps G-H substituting 2-methoxyethanol in place of 2-(1-methyl-1H-imidazol-2-yl)ethan-1-ol and deprotected according to the method of Example 2, Step I (28.0 mg, 0.0588 mmol, 68%). LCMS (MM-ES+APCI, Pos): m/z 476.2 (M+H). EXAMPLE 71
• Step A tert-butyl ( ⁇ R.5S)-3-(8-f1uoro-7-(3-hvdroxynaphthalen- l -yl)-2-(Yl - oxidotetrahvdro-2H-thiopyran-4-yl)oxy)pyrido[4.3-d]pyrimidin-4-yl )-3.8- diazabicvclo[3.2.11octane-8-carboxylate: A solution of tert-butyl (lR,5S)-3-(8-fluoro-7-(3- hydroxynaphthalen-l-yl)-2-((tetrahydro-2H-thiopyran-4-yl)oxy)pyrido[4,3-d]pyrimidin-4-yl)- 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (93 mg, 0.15 mmol) in THF (3.0 ml)
• Step B 4-(Y4-((TR.5S)-3.8-diazabicvclor3.2.noctan-3-vD-8-fluoro-7-(3- hydroxynaphthalen-1 -yl )pyridor4.3-dlpyrimidin-2-yl )oxy)tetrahvdro-2H-thiopyran 1-oxide: tert- butyl (lR,5S)-3-(8-fluoro-7-(3-hydroxynaphthalen-l-yl)-2-((l-oxidotetrahydro-2H-thiopyran-4- yl)oxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate was deprotected according to the method of Example 2, Step I (37.9 mg, 0.0498 mmol, 85%).
• LCMS MM-ES+APCI, Pos
• Step A tert-butyl ( ⁇ R.5S)-3-(7-chloro-8-f1uoro-2-methoxypyridor4.3-d1pyrimidin-4-yl )- 3.8-diazabicvclo[3.2.noctane-8-carboxylate: To a solution of tert-butyl (lR,5S)-3-(2,7-dichloro- 8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (200 mg, 0.467 mmol) in anhydrous THF (20 mL) at room temperature was added NaOMe (0.117 mL, 0.514 mmol).
• Step B tert-butyl _ (T R.5 S)-3 -(8-fluoro-7-(3 -hydroxynaphthalen- 1 -vO-2- methoxypyrido[4.3-d1pyrimidin-4-vD-3.8-diazabicvclo[3.2.
• noctane-8-carboxylate A mixture of 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)naphthalen-2-ol (93 mg, 0.344 mmol), tert- butyl (lR,5S)-3-(7-chloro-8-fluoro-2-methoxypyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (97 mg, 0.229 mmol), K2CO3 (0.344 mL, 0.688 mmol), Pd(PPh3)4 (26.5 mg, 0.023 mmol) in dioxane (2.3 mL, 0.229 mmol) was sparged with argon and heated at 85°C for 16 hours.
• Step C 4-(4-(YlR.5S)-3.8-diazabicvclor3.2.noctan-3-vD-8-fluoro-2-methoxypyridor4.3- d1pyrimidin-7-vDnaphthalen-2-ol: To a solution of tert-butyl (lR,5S)-3-(8-fluoro-7-(3- hydroxynaphthalen-l-yl)-2-methoxypyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (106 mg, 0.2002 mmol) in CH2CI2 (4 ml, 0.2002 mmol) was added TFA (0.308 mL, 4.003 mmol) at 0°C.
• Step A Synthesized according to Example 3, Steps G-H substituting (tetrahydro-lH- pyrrolizin-7a(5H)-yl)methanol in place of 2-(l-methyl-lH-imidazol-2-yl)ethan-l-ol in Step G and 2-(3-(benzyloxy)naphthalen-l-yl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane in place of 4- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)naphthalen-2-ol in Step H to afford tert-butyl ( 1 R, 5 S)-3 -(7-(3 -(benzyloxy)naphthalen- 1 -yl)-8-fluoro-2-((tetrahy dro- 1 H-pyrrolizin-7a(5H)- yl)methoxy)pyrido[4,3
• Step B utyl ( ⁇ R.5S)-3-(8-fluoro-7-(3-hvdroxynaphthalen- l-yl )-2-(Ttetrahydrol H- pynOlizin-7a(5H)-vOmethoxy)pyrido[4.3-d]pyrimidin-4-vO-3.8-diazabicvclo[3.2.noctane-8- carboxyl ate: A stirred mixture of tert-butyl (lR,5S)-3-(7-(3-(benzyloxy)naphthalen-l-yl)-8- fluoro-2-((tetrahydro-lH-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.
• Step C 4-(4-(( ⁇ R.5S)-3.8-diazabicvclo[3.2. 1 loctan-3-yl )-8-fluoro-2-(Ttetrahydro- l H- pyrrolizin-7a(5H)-yl )methoxy)pyrido[4.3-d]pyrimidin-7-yl )naphthalen-2-ol _ bis(2.2.2- trifluoroacetate): To a solution of tert-butyl (lR,5S)-3-(8-fluoro-7-(3-hydroxynaphthalen-l-yl)- 2-((tetrahydro-lH-pynOlizin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (20.3 mg, 0.032 mmol) in CH2CI2
• Step A 4.4.5.5-tetramethyl-2-(8-methylnaphthalen-l-vO-1.3.2-dioxaborolane.
• l-bromo-8-methylnaphthalene 1 g, 4.5 mmol
• dioxane 23 mL
• potassium acetate 1.3 g, 14 mmol
• 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(l,3,2-dioxaborolane) 3.4 g, 14 mmol
• PdCl2(dppf) 330 mg, 0.45 mmol.
• Step B 4-(YlR.5SV3.8-diazabicvclo[3.2.noctan-3-vO-7-(8-methylnaphthalen-l-vO-2- (Y(SVl-methylpynOlidin-2-vDmethoxy)pyrido[4.3-d1pyrimidine.
• Step A (EV 1 -bromo-8-(2-ethoxyvinv0naphthalene.
• Step B 2-(8-bromonaphthalen-l-v0ethan-l-ol.
• To a stirred solution of (£)-l-bromo-8-(2- ethoxyvinyl)naphthalene (200 mg, 0.72 mmol) in tetrahydrofuran (2.5 ml) was added cone. aq. hydrogen chloride (0.5 ml, 3.00 mmol) at once and the reaction mixture was stirred for lh.
• the mixture was partitioned between EtO Ac (20 mL) and water (10 mL) and the layers were separated. The organic layer was washed with 0.5M NaHC03 and poured into a flask.
• Step C 2-(8-bromonaphthalen-l-v0ethyl methanesulfonate.
• reaction mixture was warmed to rt during 2h, and then partitioned between hexane-EtOAc (1:1, 15 mL) and 0.5M NaHCCh (5 mL). The layers were separated. The organic phase was washed with brine, dried over Na2SC>4, and evaporated in vacuo. The residue was dissolved in MTBE (2 mL), filtered and evaporated under N2 to yield crude 2-(8-bromonaphthalen-l-yl)ethyl methanesulfonate as a colorless oil.
• Step D 3-(8-bromonaphthalen-l-vQpropanenitrile.
• a mixture of crude 2-(8- bromonaphthalen-l-yl)ethyl methanesulfonate (393 mg, 1.19 mmol), sodium cyanide (88 mg, 1.8 mmol) and N,N-dimethylacetamide (2.4 mL) was stirred at r.t. for 3 days, then heated to 50°C for 4h. The mixture was cooled and partitioned between EtO Ac (15 mL) and water (10 mL). The layers were separated. The organic phase was washed with brine, dried over Na2S04 and evaporated in vacuo. The residue was chromatographed on silica gel eluting with 10% EtO Ac/hexane to yield 3-(8-bromonaphthalen-l-yl)propanenitrile as colorless heavy oil (290 mg, 93%).
• Step E 3-(8-(4A5.5-tetramethyl- l .3.2-dioxaborolan-2-yl jnaphthalen- l -vDpropanenitrile.
• 3-(8-bromonaphthalen-l-yl)propanenitrile 74 mg, 0.28 mmol
• dioxane 1.42 mL
• potassium acetate 1.3 g, 0.85 mmol
• 4,4,4',4',5,5,5',5'-octamethyl-2,2'- bi(l,3,2-dioxaborolane) (217 mg, 0.85 mmol) and the reaction was sparged with N2 for 15 minutes followed by addition of PdCh(dppf) (21 mg, 0.028 mmol).
• the reaction was heated to 95°C for 18 hrs.
• the reaction was concentrated in vacuo and taken up in DCM.
• the slurry was filtered through GF/F paper and the organics was concentrated in vacuo.
• the material was chromatographed using 0— >30% ethyl acetate/hexane as eluent to give 3-(8-(4,4,5,5-tetramethyl- l,3,2-dioxaborolan-2-yl)naphthalen-l-yl)propanenitrile (48 mg, 0.08 mmol, 27 % yield).
• Step F 3-(8-(4-((lR.5S)-3.8-diazabicvclo[3.2.1]octan-3-vn-8-fluoro-2-(((S)-l- methylpyrrolidin-2-yl )methoxy)pyrido[4.3-d1pyrimidin-7-yl )naphthalen- l -yl ipropanenitrile bis(2.2.2-trifluoroacetate).
• Step A 8-bromo- 1 -methyl- 1.2.3.4-tetrahydronaphthalen- 1 -ol .
• the mixture was cooled in an ice bath and added dropwise to a stirred mixture of ice (50 g) and acetic acid (10 mL).
• the suspension was extracted with MTBE (100 mL).
• the organic layers were separated and washed with water, 2M Na2CCb and brine (20 mL each) and the combined water phases were reextracted with MTBE.
• the combined organic extracts were evaporated in vacuo , re-dissolved in methanol (10 mL) and 35% hydrazine (2 mL) was added.
• the reaction mixture was heated to reflux for lh.
• the reaction was cooled, partitioned between hexane and water (50 mL each) and the layers were separated.
• Step B 8-Bromo- 1 -methyl- 1.2.3.4-tetrahydronaphthalene.
• Step C 4.4.5.5-tetramethyl-2-(8-methyl-5.6.7.8-tetrahydronaphthalen-l -yl )-l .3.2- dioxab or plane.
• a stirred solution of 8-bromo-l -methyl- 1,2,3, 4-tetrahydronaphthalene (300 mg, 1.33 mmol) in tetrahydrofuran (7 mL) under N2 was cooled in a CCh-acetone bath and butyllithium (0.59 mL, 1.47 mmol) was added dropwise over 5 min.
• Step D Tert- butyl P R.5S)-3-(8-fluoro-7-(8-methyl-5.6.7.8-tetrahvdronaphthalen- l -yl )-2- (Y(SVl-methylpyrrc>lidin-2-vOmethoxy)pyrido[4.3-d1pyrimidin-4-vO-3.8- diazabicvclo[3.2.11octane-8-carboxylate.
• Step E 4-(YlR.5S)-3.8-diazabicvclor3.2.noctan-3-vO-8-fluoro-7-(8-methyl-5.6.7.8- tetrahvdronaphthalen-l-vn-2-(((S)-l-methylpyrrolidin-2-vnmethoxy)pyridor4.3-d1pyrimidine.
• the residual material was partitioned between ethyl acetate (100 mL) and water (100 mL) and the layers were separated. The organics were washed 1 x 100 mL with brine, dried over MgSCri, filtered and concentrated. The crude product was purified by flash chromatography eluting with an ethyl acetate/hexanes gradient (20% to 80% ethyl acetate).
• the crude product (5.55 g total) contained a mixture (approximately 2.7: 1) of ethyl 2-(2- (chloromethyl)allyl)-5-oxopyrrolidine-2-carboxylate and ethyl 2-methylene-5-oxotetrahydro- lH-pyrrolizine-7a(5H)-carboxylate (product of the step B) and was carried on crude without further purification.
• Step B Ethyl 2-methylene-5-oxotetrahvdro-lH-pyrrolizine-7a(5H)-carboxylate.
• NaH 139 mg, 3.47 mmol
• THF tetrahydro-lH-pyrrolizine-7a(5H)-carboxylate
• Step C Ethyl 2.5-dioxotetrahydro- l H-pyrrolizine-7a(5H)-carboxylate.
• Step D Ethyl 2-hvdroxy-5-oxotetrahvdro-lH-pyrrolizine-7a(5H)-carboxylate.
• Ethyl 2,5- dioxotetrahydro-lH-pyrrolizine-7a(5H)-carboxylate (1.08 g, 5.113 mmol) was charged to a 50 mL round bottom flask equipped with a stir bar and nitrogen inlet with methanol (17 ml, 5.1 mmol).
• sodium borohydride neat (0.14 g, 3.8 mmol).
• Step E Ethyl 2-fluoro-5-oxotetrahvdro-lH-pyrrolizine-7a(5H)-carboxylate.
• crude ethyl 2-hydroxy-5-oxotetrahydro-lH-pyrrolizine-7a(5H)-carboxylate (4.8: 1 cis:trans isomers) (1 g, 4.69 mmol) in dichloromethane (14.2 ml, 4.69 mmol) at -78 °C was added Deoxo-Fluor (0.86 ml, 4.7 mmol) neat by syringe. The reaction was stirred overnight and warmed to rt.
• the mixture was then partitioned between 25% IPA/DCM and water and the layers were separated.
• the aqueous layer was washed 3x with 25% IPA/DCM and the organics were combined and dried over Na2SC>4.
• the crude product was concentrated purified by flash chromatography eluting with an ethyl acetate/hexanes gradient (0% to 60% ethyl acetate) to yield ethyl 2-fluoro-5-oxotetrahydro-lH-pyrrolizine-7a(5H)-carboxylate as a clear oil containing a single racemic trans diastereomer (210 mg, 0.98 mmol, 21%).
• Step F (2-fluorotetrahvdro-lH-pynOlizin-7a(5H)-vDmethanol.
• Ethyl 2-fluoro-5- oxotetrahydro-lH-pyrrolizine-7a(5H)-carboxylate (.21 g, 0.990 mmol) and dry THF (2 ml) were charged to a 25 mL pear shaped flask equipped with a stir bar. The mixture was cooled to 0 °C and LAH (1M in THF) (2.97 ml, 2.97 mmol) was added dropwise. The vessel was equipped with a cold-water condenser and heated to 70 °C for 4 hours.
• Step G 4-(YlR.5S)-3.8-diazabicvclo[3.2.noctan-3-vD-7-(8-chloronaphthalen-l-vD-8- fluoro-2-(Y(2R.7aS)-2-fluorotetrahvdro-l H-pyrrolizin-7a(5H)-yl )methoxy)pyrido[4.3- dlpyrimidine was synthesized according to Example 29, Steps H-I substituting ((2R,7aS)-2- fluorotetrahydro-lH-pyrrolizin-7a(5H)-yl)methanol in place of (S)-( 1 -i sopropy 1 pyrrol i di n-2- yl)methanol (34 mg, 0.058 mmol, 62%). Submitted as a TFA salt.
• LCMS MM-ES+APCI, Pos
• Step A tert-butyl ( ⁇ R.5S)-3-(2-((3-(((tert-butyldimethylsilyl )oxy)methyl )tetrahydro- l H- pyrrolizin-7a(5H)-yl )methoxy)-7-(8-chloronaphthalen-l -yl )-8-fluoropyrido[4.3-d]pyrimidin-4- vO-3.8-diazabicvclo[3.2.noctane-8-carboxylate: To a mixture of (3-(((tert- butyldimethylsilyl)oxy)methyl)tetrahydro-lH-pyrrolizin-7a(5H)-yl)methanol (0.046 g, 0.16 mmol), tert-butyl (lR,5S)-3-(2-chloro-7-(8-chloronaphthalen-l-yl)-8-fluoropyrido[
• Step B (7a-(((4-(nR.5S)-3.8-diazabicvclo[3.2.1]octan-3-vn-7-(8-chloronaphthalen-l-vn- 8-fluoropyrido[4.3-d1pyrimidin-2-yl )oxy)methyl jhexahydro- 1 H-pyrrolizin-3-yl imethanol: Tert- butyl (lR,5S)-3-(2-((3-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-lH-pyrrolizin-7a(5H)- yl)methoxy)-7-(8-chloronaphthalen-l-yl)-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (0.015 g, 0.0187 mmol) was constituted
• Step A tert-butyl ( ⁇ R.5S)-3-trityl-3.8-diazabicvclol3.2. 1 loctane-8-carboxylate.
• tert-butyl (lR,5S)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate 1.0 g, 4.7 mmol
• triethyl amine 0.79 ml, 5.6 mmol
• Trityl-Cl 1.g, 5.0 mmol
• Step B tert-butyl 1 -methyl-3-trityl-3.8-diazabicvclor3.2. 1 loctane-8-carboxylate.
• tert-butyl (lR,5S)-3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (0.30 g, 0.66 mmol) and N,N,N',N'-tetramethylethylenediamine (0.23 ml, 1.5 mmol) in diethyl ether (4.4 ml) at - 30 °C under N2 was added sBuli (1.4 M in cyclohexane, 1.1 ml, 1.5 mmol).
• Step C tert-butyl 1 -methyl-3, 8-diazabicvclo[3.2. 1 loctane-8-carboxylate.
• HC1 1.0 N, 1.8 ml, 1.8 mmol
• the solution was stirred at rt for 1 h and was then treated with solid NaHCCh (0.15 g, 1.8 mmol). The mixture was stirred at rt for 10 min.
• Step D. 2.4; 7-tri chi pro- 8 -fluoropyri do [4.3 -d] pyrimi dine To a flask containing 7-chloro-8- fluoropyrido[4,3-d]pyrimidine-2,4(lH,3H)-dione (0.93 g, 4.3 mmol) was added POCh (8.0 ml, 86 mmol). The mixture was cooled with an ice bath and DIPEA (2.2 ml, 13 mmol) was added. The ice bath was removed, and mixture was heated between 100-110 °C until a clear solution was obtained ( ⁇ 20 h). The resulting solution was cooled and concentrated to give a brown oil.
• DIPEA 2.2 ml, 13 mmol
• Step E 4-(benzyloxy)-2.7-dichloro-8-f1uoropyridor4.3-d1pyrimidine.
• Step F (S)-4-(benzyloxy)-7-chloro-8-fluoro-2-(n-methylpyrrolidin-2- yl )methoxy)pyrido[4.3-d1pyrimidine.
• 4-(benzyloxy)-2,7-dichloro-8- fluoropyrido[4,3-d]pyrimidine (1.30 g, 4.01 mmol) in 1,4-dioxane (40 ml) was added (S)-(l- methylpyrrolidin-2-yl)methanol (0.667 ml, 5.61 mmol) followed by CS2CO3 (3.27 g, 10.0 mmol).
• Step G (S)-4-(4-(benzyloxy)-8-fluoro-2-((l -methyl pyrrol idin-2-yl )methoxy)pyrido[4.3- d1pyrimidin-7-vDnaphthalen-2-ol.
• Step H (S)-8-fluoro-7-(3-hvdroxynaphthalen-l-vD-2-(Yl-methylpyrrolidin-2- yl )methoxy)pyridor4.3-dlpyrimidin-4-ol.
• Pd/C 160 mg, 0.151 mmol.
• Step I (S)-7-(3-((tert-butyldimethylsilvnoxy)naphthalen-l-vn-8-fluoro-2-((l- m ethyl pyrrol idin-2-yl )methoxy)pyrido[4.3-d1pyrimidin-4-ol.
• Step J tert-butyl 3-(7-(3-((tert-butyldimethylsilvnoxy)naphthalen-l-vn-8-fluoro-2-(((S)- 1 -methylpynOlidin-2-vDmethoxy)pyrido[4.3-d1pyrimidin-4-vD-l -methyl-3.8- diazabicvclo[3.2.11octane-8-carboxylate.
• Step K 4-(8-fluoro-4-( 1 -methyl -3.8-diazabicvclor 3 2 11octan-3 -vO-2-(Y(S V 1 - methylpyrrolidin-2-yl )methoxy)pyridor4.3-dlpyrimidin-7-yl )naphthalen-2-ol _ bis(2.2.2- trifluoroacetate.
• Step A tert-butyl 1 -formyl -3 -trityl -3.8 -di azabi cvcl o [3.2. 1] octane- 8 -carb oxyl ate .
• Step B tert-butyl 1 -formyl-3.8-diazabicvclo[3.2. 1 loctane-8-carboxylate acetate.
• tert-butyl l-formyl-3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate 600 mg, 1.24 mmol
• acetic acid 2.5 ml
• the mixture was stirred at 50 °C for 1 h and was then concentrated to dryness. The residue was co-evaporated with heptane (5 ml c 3) to give the crude desired product as a solid.
• Step C tert-butyl ( ⁇ R.5S)-3-(2.7-dichloro-8-f1uoropyrido[4.3-d1pyrimidin-4-yl )- l -formyl - 3.8-diazabicvclo[3.2.11octane-8-carboxylate.
• Step D tert-butyl _ 3-(7-chloro-8-fluoro-2-(((S)-l-methylpyrrolidin-2- vOmethoxy)pyrido[4.3-d]pyrimidin-4-vO-l-formyl-3.8-diazabicvclo[3.2. noctane-8- carboxyl ate.
• Step E tert-butyl 3-(8-fluoro-7-(3-hvdroxynaphthalen-l-vn-2-(((S)-l-methylpyrrolidin-2- vDmethoxy)pyrido[4.3-d1pyrimidin-4-vD-l-formyl-3.8-diazabicvclo[3.2. noctane-8- carboxyl ate.
• Step F tert-butyl 1 -(2-cvanovinvD-3 -(8-fluoro-7-(3 -hydroxynaphthalen- 1 -vD-2-((YS)- 1 - methylpyrrolidin-2-yl )methoxy)pyrido[4.3-d1pyrimidin-4-yl )-3.8-diazabicvclo[3.2. 1 loctane-8- carboxylate.
• Step G tert-butyl l-(2-cvanoethvn-3-(8-fluoro-7-(3-hvdroxynaphthalen-l-vn-2-(((S)-l- methylpyrrolidin-2-yl )methoxy)pyridor4.3-dlpyrimidin-4-yl )-3.8-diazabicvclor3.2. 1 loctane-8- carboxylate.
• Step H 3-(3-(8-fluoro-7-(3-hvdroxynaphthalen-l-vn-2-(((S)-l-methylpyrrolidin-2- vDmethoxy)pyrido[4.3-d1pyrimidin-4-yr)-3.8-diazabicvclo[3.2.noctan-l-vDpropanenitrile bis(2.2.2-trifluoroacetate).
• Step A 8-(tert-butvD 1-methyl 3-trityl-3.8-diazabicvclol3.2. 1 loctane- 1.8-dicarboxylate.
• tert-butyl (lR,5S)-3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (0.45 g, 0.99 mmol) in diethyl ether (9.9 ml) at -40 °C was added N1,N1, N2,N2-tetramethylethane- 1,2- diamine (0.22 ml, 1.48 mmol), followed by slow addition of sec-butyllithium (1.06 ml, 1.48 mmol).
• Step B tert-butyl 1 -formyl-3.8-diazabicvclor3.2. 1 loctane-8-carboxylate acetate.
• 8-(tert-butyl) 1-methyl 3-trityl-3,8-diazabicyclo[3.2.1]octane-l,8-dicarboxylate 157 mg, 0.31 mmol
• HC1 1.0 N, 0.77 ml, 0.77 mmol
• the solution was stirred at rt for 1 h and was then treated with NaElCCh (77.2 mg, 0.92 mmol).
• the mixture was stirred at rt for 30 min and concentrated to dryness to give a white solid.
• the solid was extracted with DCM (10 ml) and passed through a filter plug. The filtrate was concentrated to give the crude desired product as a white solid.
• Step C 8-(tert-butvD 1-methyl 3-(7-(3-(Ytert-butyldimethylsilvDoxy)naphthalen-l-vD-8- fluoro-2-(((S)-l-methylpyrrolidin-2-vnmethoxy)pyrido[4.3-d]pyrimidin-4-vn-3.8- diazabicvclo[3.2.11octane-E8-dicarboxylate.
• Step D 8-(tert-butvD 1-methyl 3-(8-fluoro-7-(3-hvdroxynaphthalen-l-vD-2-(Y(S)-l- methylpyrrolidin-2-yl )methoxy)pyrido[4.3-d1pyrimidin-4-yl )-3.8-diazabicvclo[3.2. 1 loctane- E8-dicarboxylate.
• Step E methyl 3-(8-fluoro-7-(3-hvdroxynaphthalen-l-vn-2-(((S)-l-methylpyrrolidin-2- vDmethoxy)pyridor4.3-d1pyrimidin-4-yr)-3.8-diazabicvclor3.2.noctane-l-carboxylatebis(2.2.2- trifluoroacetate).
• Step A 8-(tert-butoxycarbonyl)-3-trityl-3.8-diazabicvclor3.2. 1 loctane- 1 -carboxylic acid.
• 8-(tert-butyl) 1-methyl 3-trityl-3,8-diazabicyclo[3.2.1]octane-l,8-dicarboxylate 131 mg, 0.26 mmol
• dioxane 5.0 ml
• NaOH 2 M, 2.2 ml, 4.4 mmol
• the mixture was heated at 80 °C for 20 h.
• the mixture was cooled to rt, acidified with citric acid (10%, 10 ml) and extracted with EtOAc.
• Step B tert-butyl 1 -carbamoyl-3-trityl-3.8-diazabicvclo[3.2. 1 loctane-8-carboxylate.
• EDC 74 mg, 0.39 mmol.
• the mixture was stirred at rt for 1 h, and NEEOEl (0.2 ml, 0.19 mmol) was added.
• Step C tert-butyl 1 -carbamoyl-3.8-diazabicvclo[3.2. 1 loctane-8-carboxylate.
• HC1 HC1 (0.20 ml, 0.20 mmol).
• the solution was stirred at rt for 1 h.
• the mixture was treated with solid NaHCCb (17 mg, 0.20 mmol) and stirred at rt for 30 min.
• the resulting mixture was concentrated to dryness to give a white solid.
• the solid was triturated with DCM (10 ml) and passed through a filter plug. The filtrate was concentrated to give the crude desired product as a white solid.
• Step D tert-butyl 3-(7-(3-((tert-butyldimethylsilvnoxy)naphthalen-l-vn-8-fluoro-2-(((S)- 1 -methylpyrrolidin-2-yl )methoxy)pyridor4.3-dlpyrimidin-4-yl )- 1 -carbamoyl-3.8- diazabicvclor 3 2 11octane-8-carboxylate.
• Step E tert-butyl l-carbamoyl-3-(8-fluoro-7-(3-hvdroxynaphthalen-l-vn-2-(((S)-l- methylpyrrolidin-2-yl )methoxy)pyrido[4.3-d1pyrimidin-4-yl )-3.8-diazabicvclo[3.2. 1 loctane-8- carboxylate.
• Step F 3-(8-fluoro-7-(3-hvdroxynaphthalen-l-vn-2-(((S)-l-methylpyrrolidin-2- vDmethoxy)pyrido[4.3-d1pyrimidin-4-yr)-3.8-diazabicvclo[3.2.noctane-l-carboxamide bis(2.2.2-trif1uoroacetate).
• Step A 4-(4-(l-chloro-3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-(((S)-l-methylpyrrolidin-2- [0565]
• Step A tert-butyl 1 -chloro-3-trityl-3.8-diazabicvclo[3.2. 1 loctane-8-carboxylate.
• Step B tert-butyl l-chloro-3.8-diazabicvclo[3.2. Hoctane-8-carboxylate.
• HC1 HC1 (0.63 ml, 0.63 mmol).
• the solution was stirred at rt for 1 h.
• the mixture was treated with solid NaElCCh (63 mg, 0.76 mmol) and stirred at rt for 30 min.
• the resulting mixture was concentrated to dryness to give a white solid.
• the solid was triturated with DCM (10 ml) and passed through a filter plug. The filtrate was concentrated to give the crude desired product as a white solid.
• Step C tert-butyl 3-(7-(3-((tert-butyldimethylsilvnoxy)naphthalen-l-vn-8-fluoro-2-(((S)- 1 -methylpyrrolidin-2-yl )methoxy)pyrido[4.3-d1pyrimidin-4-yl )- 1 -chloro-3.8- diazabicvclo[3.2.11octane-8-carboxylate.
• Step D tert-butyl 1 -chloro-3 -(8-fluoro-7-(3 -hydroxynaphthalen- 1 -vD-2-(((S)- 1 - methylpyrrolidin-2-yl )methoxy)pyridor4.3-dlpyrimidin-4-yl )-3.8-diazabicvclor3.2. 1 loctane-8- carboxylate.
• Step E 4-(4-(l-chloro-3.8-diazabicvclo[3.2.Hoctan-3-yr)-8-fluoro-2-(Y(S)-l- methylpyrrolidin-2-yl )methoxy)pyrido[4.3-d1pyrimidin-7-yl )naphthalen-2-ol _ bis(2.2.2- trifluoroacetate).
• Step A 7 -bromo- 1.1 -dimethyl-2.3 -dihydro- 1 H-indene .
• DCM 5.0 ml
• -40 °C dimethylzinc (2 M, 2.2 ml, 4.40 mmol).
• the resulting mixture was stirred at -40 and -30 °C for 20 min.
• 7-bromo-2,3-dihydro-lH-inden-l-one (422 mg, 2.00 mmol) in DCM (2.0 ml) was added dropwise.
• Step B 2-(3.3-dimethyl-2.3-dihvdro- l H-inden-4-yl )-4A5.5-tetramethyl- l .3.2- dioxaborolane.
• Step C tert-butyl (7 R.5 S)-3 -(7 -(3.3 -dimethyl-2.3 -dihydro- 1 H-inden-4-vD-8-fluoro-2- (((S)-l-methylpynOlidin-2-vDmethoxy)pyrido[4.3-d1pyrimidin-4-vD-3.8- diazabicvclor3.2.11octane-8-carboxylate 2.2.2-trifluoroacetate.
• Step D 4-(YlR.5S)-3.8-diazabicvclo[3.2. noctan-3-vO-7-(3.3-dimethyl-2.3-dihvdro-lH- inden-4-vn-8-fluoro-2-(((S)-l-methylpyrrolidin-2-vnmethoxy)pyrido[4.3-d1pyrimidine bis(2.2.2-trifluoroacetate).

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