BRPI0622030A2 - 7-SUBSTITUTED PURINE DERIVATIVES FOR IMMUNOSUPPRESSION - Google Patents

Description

"7-SUBSTITUTED PURINE DERIVATIVES FOR IMMUNOSUPPRESSION"

FIELD OF INVENTION

The invention relates to purinone derivatives useful as immunosuppressants.

BACKGROUND OF THE INVENTION

Immunosuppression is an important clinical approach in the treatment of autoimmune diseases and in the prevention of organ and tissue rejection. Clinically available immunosuppressants, including azathioprine, cyclosporine, and tacrolimus, although effective, often cause undesirable side effects, including nephrotoxicity, hypertension, gastrointestinal disorders, and gum inflammation. Jak3 tyrosine kinase inhibitors are known to be useful as immunosuppressants (see U.S. Patent 6,313,129).

Members of the Janus kinase family (Jak - Janus kinase) of intracellular non-receptor tyrosine kinases are components of cytokine signal transduction. Four family members have been identified so far: Jakl, Jak2, Jak3 and Tyk2 (Tyk - Tyrosine kinase). Jaks play a key role in cytokine receptor-mediated intracellular signaling. In binding cytokines to their receptors, Jaks are activated and phosphorylate the receptors, creating anchor sites for other signaling molecules, in particular members of the transcriptional activator and signal transducer (TSAT) family. While Jakl, Jak2, and Tyk2 expression is relatively ubiquitous, Jak3 expression is temporally and spatially regulated. Jak3 is predominantly expressed in hematopoietic lineage cells; it is constitutively expressed in natural killer cells (AN) and thymocytes and is inducible in T cells, B cells and myeloid cells (reviewed in Ortmann, et al., 1999 and Yamaoka, et al., 2004). Jak3 is also expressed in stem cells and its enzymatic activity is increased by FcsRI IgE / cross-linking receptors (Malaviya and Uckun, 1999).

A specific orally active Jak3 inhibitor, CP-690.550, has been shown to act as an effective immunosuppressant and prolonged animal survival in a heart transplant murine model and a kidney transplant primate model (Changelian, et al. , 2003).

In addition, anomalous Jak3 activity has been associated with a leukemic form of cutaneous T-cell lymphoma (Sezary's syndrome) and acute lymphoblastic leukemia (ALL), the most common form of childhood cancer. Identification of Jak3 inhibitors has provided the basis for new clinical approaches in the treatment of leukemia and lymphomas (reviewed in Uckun, et al., 10 2005). Two dimethoxyquinazoline derivatives, WHI-P131 (JANEX-I) and WHI-P154 (JANEX-2), have been reported to be selective Jak3 inhibitors in leukemic cells (Sudbeck et al., 1999).

Jak3 has also been shown to play a role in allergic reactions and mast cell-mediated inflammatory diseases, serving as a target in indications such as asthma and anaphylaxis.

Thus Jak3 inhibiting compounds are useful for indications such as leukemia and lymphomas, organ and bone marrow transplant rejection, allergic reactions and inflammatory diseases, and mast cell-mediated disorders.

SUMMARY OF THE INVENTION

It has now been found that the compounds of general formula III are potent selective inhibitors of Jak3:

In these compounds,

Qi and Q2 are independently selected from the group consisting of CX1, CX2 and nitrogen, where Q1 and Q2 are not both nitrogen;

Q3 is N or CH;

X1 and X2 are independently selected from the group consisting of hydrogen, (C1-C6) alkyl, cyano, halo, (C1-C6) alkyl, hydroxyl, (C1-C6) alkoxy; halo (C1 -C6) alkoxy and nitro;

R1 is selected from the group consisting of hydrogen and (C1-C6) alkyl;

y is zero or an integer selected from 1, 2 and 3;

R2 and R3 are independently selected for each occurrence of (CR2R3) from the group consisting of hydrogen and (C1-C6) alkyl;

R4 is selected from a group consisting of alkyl, heterocyclyl, aryl, heteroaryl, substituted alkyl, substituted heterocyclyl, substituted aryl, substituted heteroaryl; and

R5 is selected from the group consisting of alkyl, heterocyclyl, substituted heterocyclyl and C1-C6 alkyl wherein

(a) one or two CH 2 are substituted by a group selected from NH and N (alkyl);

(b) one or two CH2 are replaced by O;

(c) one or two CH 2 are replaced by (C = O);

(d) two CH2 are replaced by CH = CH or C = C;

or

(e) any chemically stable combination of (a), (b) (c) and (d);

and wherein from zero to three hydrogens are replaced by a substituent chosen from:

(a) halogen, hydroxy, cyano, lower alkylsulfonyl, lower alkylsulfonyloxy, amino, lower alkylamino, lower dialkylamino, alkoxyamino, sulfonylamino, acylamino, arylamino, lower alkoxy;

(b) heterocyclyl and heterocyclyl substituted with

one to three substituents chosen from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl;

(c) phenyl and phenyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl, acylamino, cyano, carboxy, alkoxycarbonyl, haloalkyl and heterocyclyl; and (d) heteroaryl and heteroaryl substituted with one to three substituents chosen from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl.

Members of these genera are useful for inhibiting Jak3 activity and as such are useful in indications where clinical immunosuppression is desired and in the treatment of hematologic cancers. The compounds are more selective for Jak3 than for Aurora A kinases, which are the corresponding compounds in which R5 is hydrogen.

In another aspect, the invention relates to

pharmaceutical compositions comprising a therapeutically effective amount of at least one compound of general formula III, or a pharmaceutically acceptable salt; and a pharmaceutically acceptable carrier.

In another aspect, the invention relates to a

method for treating a disease by altering a Jak3 tyrosine kinase-mediated response. The method comprises contacting Jak3 with at least one compound of the general formula III.

In yet another aspect, the present invention relates to a method of suppressing the immune system in a subject in need, comprising administering to the subject a therapeutically effective amount of at least one compound of general formula III.

In a further aspect, the present invention relates to a method for treating a selected disease or disorder of an autoimmune disease, an inflammatory disease, a mast cell-mediated disease, a hematological malignancy and organ transplant rejection, in particular. a subject in need, comprising administering to a subject a therapeutically effective amount of at least one compound of general formula III.

Suppression of immune system activity is desirable to prevent or treat organ or tissue rejection following transplant surgery and to prevent and treat diseases and disorders arising from aberrant immune system activity, in particular autoimmune disorders and diseases. Exemplary autoimmune disorders include graft versus host disease (GVHD), insulin-dependent diabetes (Type I), Hashimoto's thyroiditis and Graves' disease, pernicious anemia, Addison's disease, chronic active hepatitis, Crohn's disease, ulcerative colitis , rheumatic arthritis, multiple sclerosis, systemic lupus erythematosus, psoriasis, scleroderma and myasthenia gravis.

The compounds of the present invention are useful in preventing and treating diseases and disorders related to mast cell-mediated allergic reactions and inflammation, such as keratoconjunctivitis sicca.

Other indications in which Jak3 inhibitors are useful include leukemias and lymphomas.

DETAILED DESCRIPTION OF THE INVENTION

Throughout this report they will be defined when first mentioned and will retain their definitions.

In a first aspect the invention relates to purinones and imidazopyridinones having the general formula III:

Members of genus III can be conveniently divided into subgenres, based on Q values. When Qi is nitrogen and Q2 is carbon, a subgenus of purinones and imidazo [4,5-b] pyridinones having an imidazo [4,5- c] pyridine attached. When Qi is carbon and Q2 is nitrogen, a subgenus of purinones and imidazo [4,5-b] pyridinones have an attached imidazo [5,4-c] pyridine. When Qi and Q2 are both carbon, a subgenus of purinones and imidazo [4,5-b] pyridinones have an attached benzimidazole. The gender could be similarly divided based on Q3. When Q3 is nitrogen, a purinone subgenus appears having an attached imidazo [4,5-c] pyridine, imidazo [5,4-c] pyridine, or benzimidazole. When Q3 is carbon, a subgenus of imidazo [4,5-b] pyridinones appears having an attached imidazo [4,5-c] pyridine, imidazo [5,4-c] pyridine, or benzimidazole. The structures of these subgenres are shown below:

10

In certain embodiments, X1 and X2 are selected from hydrogen, cyano, chlorine, fluorine, trifluoromethyl, trifluoromethoxy and methyl; in other embodiments R1 is H. In a subdivision, y is zero; in another y is 1 or 2 and R2 and R3 are hydrogen or methyl. Examples of R4 include: cyclopentyl, cyclohexyl, piperidine, oxepane, benzoxepane, di-

hydrocyclopentapyridine, phenyl, benzyl, tetraline, indane, tetrahydropyran, tetrahydrofuran, tetrahydroindole,

isoquinoline, tetrahydroisoquinoline, quinoline, tetrahydro-

hydroquinoline, chroman, pyridine, pyrimidine, pyrazine, dihydropyran, dihydrobenzofuran, tetrahydrobenzofuran, tetrahydrobenzothiophene, furan, dihydropyran [2,3-b] pyridine (see example below), tetrahydroquinoxaline, tetrahydrothiopyran (thian), thiochroman (dihydrobenzothiine), or any of the preceding rings bearing 1 to 3 additional substituents such as halogen, methyl, methoxy, trifluoromethyl, cyano, hydroxy, oxo, 5 oxide and acetyl. Additional examples in which R4 is alkyl or substituted alkyl include subgenera in which R4 is oxaalkyl (alkoxyalkyl).

In certain embodiments of genre III, y is 1 or 2; R 2 and R 3 are hydrogen or methyl and R 4 is phenyl, quinoline, pyridine, pyrazine or substituted phenyl, quinoline, pyridine or pyrazine. In other embodiments of genus III, y is zero and R 4 is cyclopentyl, cyclohexyl, phenyl, indane, piperidine, oxepane, benzoxepane, dihydrocyclopentapyridine, tetraline, tetrahydropyran, tetrahydrofuran, tetrahydroindola , isoquinoline, tetrahydroisoquinoline, quinoline, tetrahydroquinoline, chroman, pyridine, pyrimidine, dihydropyran, dihydrobenzofuran, tetrahydrobenzofuran, tetrahydrobenzothiophene, furan, dihydropyran [2,3-b] pyridine tetrahydroquinoxaline, tetrahydrothiopyran (thian), thiochroman (dihydrobenzothiine), or a substituted ring from the preceding list. In additional embodiments, (a) y is zero and R 4 is selected from cyclohexyl, oxepane, tetraline, indane, dihydrocyclopentapyridine, tetrahydropyran, tetrahydroquinoline, chroman, dihydrobenzofuran, tetrahydrobenzofuran, di -hydropyran [2,3-b] pyridine and tetrahydroquinoxaline, each optionally substituted with hydroxy, oxo, or halogen; or (b) y is 1 or 2, R 2 and R 3 are hydrogen or methyl and R 4 is selected from phenyl, pyridine and pyrazine, each optionally substituted with halogen. When y is zero, R 4 may be tetrahydropyran-4-yl, 4-hydroxycyclohexyl, 4-oxocyclohexyl, oxepan-4-yl, chroman-4-yl; 3,4-di-

hydronaphthalene-1 (2H) -one-4-yl; 2,3-dihydro-inden-1-one-4-yl and its substituted fluorine counterparts. It appears that although both enantiomers are active, the compounds in which the chroman carbon at position 4 is of the (R) configuration have higher potency. Certain preceding subgenres in which y is zero can also be described by a representation in which R4 is: According to this representation, W is CH2, C = O or O; p is 1, 2 or 3; and A is a six membered heteroaromatic ring containing 1 or 2 nitrogen, or a benzene ring optionally substituted with one or two fluorines. The wavy line denotes the point of attachment to the purinone. Compounds in which carbon marked with an asterisk

of configuration (R), appear to be more potent than their corresponding (S) enantiomers. Other embodiments include compounds wherein y is I and R 4 is selected from difluorophenyl, fluorophenyl, chlorophenyl, chlorofluorophenyl, pyridin-3-yl and pyrazin-3-yl.

In certain embodiments of genus III, Xi and X2 are selected from hydrogen, cyano, chlorine, fluorine, trifluoromethyl, trifluoromethoxy and methyl. In more limited embodiments, X 1 is selected from hydrogen, cyano and fluorine; Q1 and Q2 are CXi; Q3 is N and R1 is H. In some embodiments, y is zero and R4 is selected from cyclohexyl, tetraline, indane, oxepane. dihydrocyclopentapyridine, tetrahydropyran, tetrahydroquinoline, chroman, dihydrobenzofuran, tetrahydrobenzofuran, dihydropyran [2,3-b] pyridine and tetrahydroquinoxaline, each optionally substituted with hydroxy, oxo, or halogen. In other embodiments y is 1 or 2, R 2 and R 3 are hydrogen or methyl and R 4 is selected from phenyl, pyridine and pyrazine, each optionally substituted with halogen. In certain embodiments, R 5 is C 1 -C 6 alkyl wherein certain substitutions and substitutions have been made. In one embodiment, (a) one or two CH 2 may be substituted by NH or N (alkyl). These wastes are also

called azaalkyl. An example of such R5 is

CH 2 CH 2 CH 2 N (CH 3) CH 2 CN, which may be considered 5-cyanopentane in which a CH 2 has been replaced by N (CH 3). In one embodiment, (b) one or two CH 2 may be substituted for O. These residues are also called oxaalkyl. In one embodiment, (c), one or two CH 2 may be replaced by (C = O). An example of such R5 is:

morpholinyl) ethyl in which CH 2 was replaced by C = O. In one embodiment, (d), two CH2 are replaced by CH = CH or C = C. An example of such R 5 is 3-methylbut-2-eno-1-yl [-CH 2 CH = C (CH 3) 2]. A further embodiment includes (e) any chemically stable combination of (a), (b) (c) and (d). An example of such R 5 is -CH 2 CH 2 CH 2 NHC (= 0) CH 2 CN, which may be considered 6-cyanohexane in which CH 2 was replaced by NH and a CH 2 was replaced by C = 0. The term "chemically stable" is readily understood by those skilled in the chemical field. It encompasses compounds that can be isolated and purified and which do not measurably decompose to physiological pH, aqueous solution, or do so at a rate that> 99% will remain after one hour at room temperature. In all of these compounds, from zero to three hydrogens may be substituted by a substituent chosen from:

(a) halogen, hydroxy, cyano, lower alkylsulfonyl, lower alkylsulfonyloxy, amino, lower alkylamino, lower dialkylamino, alkoxyamino, sulfonylamino, acylamino, arylamino, lower alkoxy; (b) heterocyclyl and heterocyclyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl;

(c) phenyl and phenyl substituted with one to three substituents chosen from halogen, hydroxy, alkoxy, alkyl, acylamino, cyano, carboxy, alkoxycarbonyl, haloalkyl and heterocyclyl.

Some examples of the above mentioned substituents for C1 -C6 alkyl include, but are not limited to:

-Ç!

/ ·

The s.

'cr Il ch3

'N

CH3

CH3

THE

OMe In certain embodiments of genus III, Q1 is CX1, Q2 is CX2, X1 is hydrogen, X2 is a substituent at the position

6 of benzimidazole and X 2 is chosen from hydrogen, fluorine and cyano.

All compounds falling within the preceding matrix genre and its subgenera are useful as Jak3 inhibitors.

Definitions

For convenience and clarity, certain terms used in this report, examples and claims are described herein.

Under the term alkyl is meant

cyclic, linear, or branched hydrocarbon structures and combinations thereof. Lower alkyl refers to alkyl groups having

1 to 6 carbon atoms. Examples of lower alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, s- and t-butyl and the like. Preferred alkyl groups are those with C20 or below; most preferred are C1-CS alkyl. Cycloalkyl is a subset of alkyl and includes cyclic hydrocarbon groups of 3 to 8 carbon atoms. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl and the like.

C1 to C20 hydrocarbons include alkyl, cycloalkyl, alkenyl, alkynyl, aryl and combinations thereof. Examples include phenethyl, cyclohexylmethyl, camphoryl and naphthylethyl.

Alkoxy or alkoxy refers to groups of 1 to 8 25 carbon atoms of a cyclic, straight, or branched configuration and combinations thereof, attached to the matrix structure by means of an oxygen. Examples include methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclohexyloxy and the like. Lower alkoxy refers to groups containing one to four carbons. The meaning of the term oxaalkyl is that used by those skilled in the art [see "Naming and Indexing of Chemical Substances for Chemical Abstracts", published by the American Chemical Society. , § 196, but without the restriction of §127 (a)], that is, 5 refers to compounds in which oxygen is bound, via a single bond, to their adjacent atoms (forming ether bonds); It does not refer to doubly bound oxygen, as would be found in carbonyl groups.

Acyl refers to groups of 1 to 8 carbon atoms of a straight, branched, saturated, unsaturated and aromatic cyclic configuration and combinations thereof, attached to the matrix structure by a carbonyl functionality. One or more carbons in the acyl residue may be replaced by nitrogen, oxygen or sulfur as long as the point of attachment to the matrix structure remains in the carbonyl. Examples include acetyl, benzoyl, propionyl, isobutyryl, t-butoxycarbonyl,

benzyloxycarbonyl and the like. Lower acyl refers to groups containing one to four carbons.

Aryl and heteroaryl means a 5- or 6-membered aromatic or heteroaromatic ring containing 0-3 heteroatoms selected from O, N, or S; a 9 or 10 membered bicyclic aromatic or heteroaromatic ring system containing 0-3 heteroatoms selected from O, N, or S; or a 13- or 14-membered tricyclic aromatic or heteroaromatic ring system, containing 0-3 heteroatoms selected from O, N, or S. The 6- to 14-membered aromatic carbocyclic rings include, for example, benzene and naphthalene and for purposes of the present invention, fused halves such as tetrahydronaphthalene (tetraline), indane and fluorine, in which one or more rings are aromatic but not all need to be. The 5- to 10-membered aromatic heterocyclic rings include, for example, imidazole, pyridine, indole, thiophene, benzopyran, thiazole, furan, benzimidazole, quinoline, isoquinoline, quinoxaline, pyrimidine, pyrazine, tetrazole and pyrazole.

Arylalkyl refers to a substituent in which

an aryl residue is attached to the matrix structure through alkyl. Examples are benzyl, phenethyl and the like. Heteroarylalkyl refers to a substituent in which a heteroaryl residue is attached to the matrix structure through alkyl. Examples include, for example, pyridinylmethyl, pyrimidinylethyl and the like.

Heterocycle means a cycloalkyl in which

one to three carbons are substituted by a hetero atom selected from the group consisting of N, O and S. Nitrogen and sulfur heteroatoms may optionally be oxidized. In some contexts (other than the present) the term heterocycle may be interpreted as including heteroaryl; For purposes of this application, heterocycle is a saturated heterocycle and does not include heteroaryl. When heteroaryl is desired, it is expressly indicated. Examples of heterocycles include pyrrolidine, morpholine, dioxane, tetrahydrofuran and the like. Examples of heterocyclyl residues additionally include piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 4-piperidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, quinuclidinyl, tetrahydrofuryl, tetrahydropyrinyl, tetrahydrofuryl, thiomorpholinyl and thiomorpholinyl sulfone. A nitrogenous heterocycle is a heterocycle containing at least one ring nitrogen; It may contain additional nitrogen as well as other hetero atoms.

Substituted alkyl, aryl, cycloalkyl, heterocyclyl, heteroaryl, etc. refers to alkyl, aryl, cycloalkyl, heterocyclyl, or heteroaryl, wherein up to three H atoms in each residue are substituted with halogen, haloalkyl, hydroxy, lower alkoxy, hydroxy lower alkyl, carboxy , carboalkoxy (also called alkoxycarbonyl), carboxamido (also called alkylaminocarbonyl),

heterocyclylcarbonyl, cyano, carbonyl, nitro, amino, alkylamino, dialkylamino, lower alkoxyamino,

arylaminocarbonyl, mercapto, alkylthio, sulfoxide, amino sulfoxide, sulfone, acylamino, amidino, alkenyl, cycloalkyl, phenyl, substituted phenyl, benzyl, substituted benzyl, heteroaryl, phenoxy, benzenesulfonyl, benzyloxy, or heteroaryloxy. Where the matrix is a heterocycle that allows such substitution, the term also includes oxides, for example pyridine-N-oxide, thiopyran sulfoxide and thiopyrane-S, S-dioxide. As mentioned above, two hydrogens on a single carbon may be substituted by one carbonyl to form an oxo derivative. Noteworthy oxo-substituted aryl residues include tetralone (3,4-dihydronaphthalene-1 (2H) -one) and indanone (2,3-dihydroinden-1-one).

The terms "halogen" and "halo" refer to fluorine, chlorine, bromine or iodine.

Some of the compounds described herein may contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers and other stereoisomeric forms which may be defined, in terms of absolute stereo-chemistry, as (R) or (S). The present invention is intended to include such possible isomers as well as their racemic and optically pure forms. Optically active (R) and (S) isomers may be prepared using chiral syntones or chiral reagents, or resolved using conventional techniques. Where the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry and, unless otherwise specified, the compounds are intended to include both geometric isomers E and Z. It is also intended that all tautomeric forms be included. . The configuration of any carbon-carbon double bond appearing herein has been elected for convenience only and is not intended to designate a particular configuration; thus, a carbon-carbon double bond described arbitrarily herein as trans can be Z, E, or a mixture of the two in any proportion.

Graphical representations of racemic, ambiscalemic and scalemic or enantiomerically pure compounds used herein are taken from Maehr J. Chem., Ed. 62, 114-120 30 (1985); solid and dashed "wedges" (triangles) are used to denote the absolute configuration of a chiral element; the wavy lines indicate the negation of any stereo-chemical implications that the bond representing it may generate; solid and dashed thick lines are geometric descriptors that indicate the relative configuration shown but denoting racemic character; and "wedge" contours (triangles) and dotted or dashed lines denote enantiomerically pure compounds of undetermined absolute configuration.

It will be appreciated that the compounds of this invention may exist in radiolabelled form, that is, the compounds may contain one or more atoms containing an atomic mass or mass number other than the atomic mass or mass number normally found in nature. Hydrogen, carbon, phosphorus, fluorine, chlorine and iodine radioisotopes include 3H, 14C, 35S, 18F, 36Cl and 125I, respectively. Compounds containing such Radioisotopes and / or other Radioisotopes of other atoms are within the scope of this invention. Tritiated radioisotopes, i.e. 3H and carbon-14, i.e. 14C, are particularly preferred because of their ease of preparation and detectability. The radiolabelled compounds of this invention may generally be prepared by methods well known to those skilled in the art. Conveniently, such radiolabelled compounds may be prepared by following the procedures described in the Examples by substituting a readily available radiolabelled reagent for an unlabeled reagent. Because of the high affinity for the active site of the Jak3 enzyme, the radiolabelled compounds of the invention are useful for Jak3 assays.

In one embodiment, R 4 is a heterocycle. Heterocycles appearing in the examples are monocyclic and bicyclic heterocycles, or monocyclic and bicyclic heterocycles substituted with one or two substitutions. When y is not zero, heteroaryl is a preferred subset of heterocyclyl for R4. Exemplary nitrogenous heterocycles include piperidine, pyridine, pyrazine, pyrimidine, pyridine, quinoline, isoquinoline, tetrahydroquinoline, tetrahydroisoquinoline and their variously substituted derivatives, such as: NH

An oxygen heterocycle is a heterocycle that contains at least one ring oxygen; It may contain additional oxygen as well as other hetero atoms. Exemplary oxygenated heterocycles include tetrahydropyran, chroman and variously substituted derivatives thereof, such as:

Ti Chemical Synthesis

The terminology related to "protect", "unprotect", and "protected" features occurs throughout this request. Such terminology is well understood by those skilled in the art and is used in the context of processes involving sequential treatment with a series of reagents. In that context, a protecting group refers to a group that is used to mask functionality during a process step in which it would otherwise react but in which the reaction is undesirable. The protecting group prevents reaction at that stage, but may subsequently be removed to expose the original functionality. Removal or "deprotection" happens after the completion of the reaction or reactions in which functionality would interfere. Thus, when a sequence of reagents is specified, as occurs in the processes of the invention, one skilled in the art can readily predict which groups would be suitable as "protecting groups". Suitable groups for this purpose are discussed in standard textbooks in the field of chemistry, such as "Protective Groups in Organic Synthesis" by T.W. Greene [John Wiley & Sons, New York, 1991], which is incorporated herein by reference.

A comprehensive list of abbreviations used in organic chemistry appears in the first issue of each volume of the Journal of Organic Chemistry. The list, which is commonly presented in a table titled "Standard Abbreviation List", is incorporated herein by reference.

In general, the compounds of the present invention may be prepared by the methods illustrated in the general reaction schemes, such as those described below, or by modifications thereof, using readily available starting materials and reagents and conventional synthesis procedures. In these reactions, it is also possible to make use of variants which are themselves known but not mentioned here. Starting materials, for example in the case of suitably substituted benzimidazole ring compounds, are commercially available, synthesized as described in the examples, or may be obtained by methods well known to those skilled in the art. The present invention further provides pharmaceutical compositions comprising as active agents the compounds described herein.

As used herein, a "composition

"Pharmaceutical" refers to a preparation of one or more compounds described herein, or physiologically acceptable salts or solvents thereof, with other chemical components such as physiologically suitable carriers and excipients.

Pharmaceutical compositions for use in accordance with the present invention may thus be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate the processing of active compounds into preparations which may be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.

Jak-3 inhibiting compounds may be formulated as pharmaceutical compositions and administered to a mammalian subject, such as a human patient, in a variety of ways adapted to the chosen route of administration, that is, orally or parenterally, intravenously, intramuscular, topical, transdermal or subcutaneous.

For oral administration, the compounds may be readily formulated by combining the active compounds with pharmaceutically acceptable carriers known in the art. Such carriers allow the compounds of the invention to be formulated as tablets, pills, pills, capsules, liquids, gels, syrups, dregs, suspensions and the like for oral ingestion by a patient. Pharmacological preparations for oral use may be made using a solid excipient, optionally milling the resulting mixture and processing the granule mixture after addition of suitable auxiliaries, if desired, to obtain tablets or dredge cores. Satisfactory excipients are in particular fillers, such as sugars, including lactose, sucrose, mannitol, or sorbitol; Cellulose preparations such as, for example, corn gum, wheat gum, rice gum, potato gum, gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carbomethylcellulose; and / or physiologically acceptable polymers such as polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added such as polyvinylpyrrolidone, agar or cross-linked alginic acid, or a related salt such as sodium alginate.

Furthermore, enteric coating may be useful as it may be desirable to prevent exposure of the compounds of the invention to the gastric environment.

Orally usable pharmaceutical compositions include push-fit capsules made of gelatin as well as soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. Push-fit capsules may contain the active ingredients mixed with a filler such as lactose, binders such as gums, lubricants such as talc or magnesium stearate and optionally stabilizers.

In soft capsules, the active compounds may be dissolved or suspended in suitable liquids such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for the chosen route of administration.

For injection, the compounds of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers, such as a Hank or Ringer's solution, or a physiological saline buffer. For transmucosal and transdermal administration, penetrants suitable for the barrier to be penetrated may be used in the composition. Such penetrants, including, for example, DMSO or polyethylene glycol, are known in the state of the art.

For administration by inhalation, the compounds for use according to the present invention are conveniently distributed in the form of an aerosol spray presentation, from a pressurized tube or nebulizer, using a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, or carbon dioxide. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to dispense a metered amount. Capsules and cartridges of, for example, gelatin, may be formulated for use in an inhaler or insufflator, containing a combination powder mixture and a suitable powder base such as lactose or gum.

Pharmaceutical compositions for parenteral administration include aqueous solutions of the active ingredients in water-soluble form. In addition, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Satisfactory lipophilic solvents or carriers include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate, triglycerides or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol or dextran. Optionally, the suspension may also contain satisfactory stabilizers or agents that increase the solubility of the compounds to allow the preparation of highly concentrated solutions.

The compounds of the present invention may also be formulated in rectal compositions, such as suppositories or retention enemas, using, for example, conventional suppository bases such as cocoa butter or other glycerides.

Depending on the severity and sensitivity of the condition to be treated, the dosage may also be in a single administration of a slow release composition, with a course of treatment lasting from several days to several weeks, or until healing is performed or decrease in disease status is achieved. The amount of a composition to be administered will, of course, depend on many factors, including the individual to be treated, the severity of the disease, the manner of administration and the judgment of the prescribing physician. The compounds of the invention may be administered orally or by injection at a dose of 0.001 to 2500 mg / kg per day. The dosage range for human adults is generally 0.005 mg to 10 g / day. Tablets or other presentations provided in separate units may conveniently contain an amount of the compound of the invention that is effective in such dosage, or as a multiple thereof, for example units containing 5 mg to 500 mg, usually around 10 mg to 200 mg. The precise amount of compound administered to a patient will be the responsibility of the attending physician. However, the dose employed will depend on several factors, including the age and gender of the patient, the disorder being treated and its severity. Also, the route of administration may vary depending on the disease and its severity.

As used herein and as would be understood by one of ordinary skill in the art, the term "a compound" is intended to include salts, solvates and inclusion complexes of that compound. The term "solvate" refers to a solid-state compound of formula I or II, wherein the molecules of a suitable solvent are incorporated into the crystalline lattice. A solvent suitable for therapeutic administration is physiologically tolerable at the dosage administered. Examples of solvents suitable for therapeutic administration are ethanol and water. When the solvent is water, the solvate is called hydrate. In general, solvates are formed by dissolving the compound in the appropriate solvent and isolating the solvate by cooling, or using an anti-solvent. The solvate is commonly dried or azeotroped under ambient conditions. Inclusion complexes are described in Remington: "The Science and Practice of Pharmacy", 19th Ed. (1995), volume 1, pages 176-177, which is incorporated herein by reference. The most commonly employed inclusion complexes are those with cyclodextrins and all natural and synthetic cyclodextrin complexes are specifically encompassed in the claims.

The term "pharmaceutically acceptable salt" refers to salts prepared from pharmaceutically acceptable non-toxic acids or bases, including inorganic acids and bases and organic acids and bases. Where the compounds of the present invention are basic, salts may be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Suitable pharmaceutically acceptable acid addition salts for the compounds of the present invention include acetic acid, benzene sulfonic (besylate), benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic acid , maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic and the like. Where the compounds contain an acidic side chain, suitable pharmaceutically acceptable base addition salts for the compounds of the present invention include metal salts made of aluminum, calcium, lithium, magnesium, potassium, sodium and zinc, or organic salts made of lysine, N, N '- dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine.

The term "prevent" as used herein refers to administering a medicament previously to prevent or minimize an attack. One of ordinary skill in the medical field (to which the present method claims are directed) should recognize that the term "prevent" is not an absolute term. In the medical field, the term refers to the prophylactic administration of a drug to substantially decrease the likelihood or severity of a pathological condition and this is the meaning intended herein.

It should be understood that, in addition to the ingredients particularly mentioned above, the formulations of this invention may include other conventional agents in the art, considering the type of formulation in question, for example those suitable for oral administration may include flavoring agents.

The compositions may be presented in a pack or dispenser, which may contain one or more unit dosage forms containing the active ingredient. Examples of a package include metal or plastic blades such as blisters and inhalation nebulizers. The pack or dispenser may be accompanied by administration instructions. Compositions comprising a compound of the present invention formulated in a compatible pharmaceutical carrier may also be placed in an appropriate container labeled for the treatment of an indicated pathological condition.

Indications

The compounds of the present invention are useful

inhibiting Jak3 activity or inhibiting Jak3 mediated activity and are useful as immunosuppressive agents for tissue and organ transplants including bone marrow transplantation and in the treatment of inflammatory and autoimmune diseases and complications arising therefrom.

Acute, chronic, and hyper-acute organ transplant rejection can be treated. Hyperacute rejection occurs minutes after transplantation. Acute rejection usually happens within six to twelve months after transplantation. Hyperacute and acute rejections are commonly reversible when treated with immunosuppressive agents. Chronic rejection, 10 characterized by gradual loss of organ function, is a continuing concern for transplant recipients because it can occur at any time after transplantation.

There are approximately 75 different known autoimmune disorders, which can be classified into two types, organ-specific (directed primarily at one organ) and non-organ-specific (affecting multiple organs).

Examples of organ-specific autoimmune disorders are insulin-dependent diabetes (Type I), which affects the pancreas, Hashimoto's thyroiditis, and Graves' disease, which affect the thyroid gland, pernicious anemia, which affects the stomach, Cushing's disease and Addison's disease affecting the adrenal glands, chronic active hepatitis affecting the liver, polycystic ovary syndrome (PCOS), celiac disease, psoriasis, inflammatory bowel disease (IBD) and 25 ankylosing spondylitis.

Examples of non-organ-specific autoimmune disorders are rheumatic arthritis, multiple sclerosis, systemic lupus, and myasthenia gravis.

Type I diabetes results from the selective aggression of self-reactive T cells against insulin-secreting β cells from the Langerhans islets. When Jak3 is the target in this disease, this is based on the observation that multiple cytokines signaling via the Jak pathway are known to participate in T-cell-mediated autoimmune destruction of 35 β cells. Indeed, it has been shown that JANEX-1, a Jak3 inhibitor, would prevent the development of spontaneous autoimmune diabetes in the NOD rat model of type I diabetes. Graft-versus-host disease (GVHD) is a pathological condition. donor T-cells that often accompany allogeneic bone marrow transplantation (BMT). Clinical and essentially experimental research 5 have shown that donor T cells are the main mediators and promoters of GVHD. Jak3 plays a key role in the induction of GVHD and treatment with a Jak3 inhibitor, JANEX-I, has been shown to attenuate the severity of GVHD (reviewed in Cetkovic-Cvrlje and Ucken, 2004).

Mast cells reveal the presence of Jak3 and Jak3 is a

fundamental regulator of IgE-mediated mast cell responses, including the release of inflammatory mediators. Jak3 has been shown to be a valid target in the treatment of mast cell mediated allergic reactions.

Allergic disorders associated with activation of

Mast cells include immediate Type I hypersensitivity reactions such as allergic rhinitis (hay fever), allergic urticaria (urticaria), angio-edema, allergic asthma and anaphylaxis, ie "anaphylactic shock". These disorders are treated or prevented by inhibiting Jak3 activity, for example by administering a Jak3 inhibitor according to the present invention.

In accordance with the present invention, Jak3 inhibitors may be administered prophylactically, that is, before the onset of an acute allergic reaction, or they may be administered after the onset of the reaction, or in both cases.

Tissue and organ inflammation occurs over a wide range of disorders and diseases, and in certain variations is the result of activation of the cytokine receptor family. Exemplary inflammatory disorders associated with Jak3 activation include, but are not limited to, skin inflammation due to radiation exposure, asthma, allergic inflammation, and chronic inflammation.

Jak3 inhibitors of the present invention also

They are useful in treating certain malignancies, including skin cancer and hematological malignancies such as lymphomas and leukemias. The following examples will further describe the invention and will be used for purposes of illustration only and should not be construed as limiting the invention described.

EXAMPLES

The following abbreviations and terms have the following

indicated meaning:

Ac = acetyl Bu = butyl

DCM = dichloromethane = methylene chloride = CH 2 Cl 2 DEAD = Diethyl Azodicarboxylate

DIC = Diisopropylcarbodiimide DIEA = N, N-Diisopropylethyl Amine DMF = N, N-Dimethylformamide DMSO = Dimethyl Sulfoxide EA (EtOAc) = Ethyl Acetate

GC = Gas Chromatography h = hours

HOAc = Acetic Acid HOBt = Hydroxybenzotriazole Me = Methyl

Pd (dppf) 2Cl 2 = dichloro [1,1'-

bis (diphenylphosphinoferrocene] palladium

Ph = Phenyl PhOH = Phenol TA = Ambient Temperature

sat = saturated s- = secondary t- = tertiary

TBDMS = t-butyldimethylsilyl TFA = Trifluoroacetic acid

THF = Tetrahydrofuran TMOF = Trimethyl orthoformate TMS = Trimethylsilyl tosyl = p-toluenesulfonyl Trt = Triphenylmethyl

Examples 1-15 describe the syntheses of certain precursors and intermediates of the invention. Example I: Synthesis of 3,4-Diaminobenzonitrile A solution of 4-amino-3-nitrobenzonitrile (1) (3.0 g) in ethanol (80 mL) was sprayed for 5 minutes with nitrogen. Palladium on carbon (10%, 300 mg) was added and the mixture was saturated with hydrogen. The mixture was stirred under a hydrogen balloon for seven hours. The mixture was sparged with nitrogen and filtered through celite. The filtrate was concentrated in vacuo to afford the title compound, 3,4-diaminobenzonitrile (2).

Example 2: Synthesis of 3H-Benzo [d] imidazole-5-

carbonitrile

A mixture of 3,4-diaminobenzonitrile (2) (1.0 g) and (ethoxymethylene) malononitrile (1.4 g) was refluxed in 50 mL of isopropyl alcohol for 16 h. The mixture was concentrated in vacuo to afford the title compound, 3H-benzo [d] imidazole-5-carbonitrile (3).

Example 3: Synthesis of 6- (Trifluoromethoxy) -H-

benzo [d] imidazola

4

6- (trifluoromethoxy) -ΙΗ-benzo [d] imidazole (4) was prepared in two steps from 2-nitro-4- (trifluoromethoxy) aniline (5) using procedures identical to those used to make 3H-benzo [d ] imidazole-5-carbonitrile (3) from 4-amino-3-nitrobenzonitrile ((1), examples 1,2).

Example 4: Synthesis of 5,6-Difluoro-IH-

benzo [d] imidazole Na2S2O4 Ργ ^ γΝΗ2 EtOCH ^ C (CN) 2 Ρ'γίΓχγ-1 \>

F-Î »siIVIVO2 NaHCO3 iPrOH

THF / H2O /

6 MeOH 7

A solution of 4,5-difluoro-2-nitroaniline (6) (1.0 g) in 30 mL of THF was treated with a solution comprising

6 g Na 2 S 2 O 4 and 3 g NaHCO 3 in 30 mL water. Methanol (10 mL) was added after addition of the aqueous solution, so that the mixture remained homogeneous. The mixture was stirred for two hours and then diluted with 100 mL ethyl acetate and 100 mL water. The organic layer was separated and the aqueous layer was extracted again with 100 mL of methylene chloride. The combined organic layers were dried over sodium sulfate, filtered and concentrated to afford crude intermediate 4,5-difluorobenzene-1,2-diamine (7). The intermediate was refluxed with (ethoxymethylene) malononitrile (1.1 g) in 25 mL of isopropyl alcohol for 16 h. The mixture was concentrated in vacuo and the resulting crude product was suspended in water and filtered. The precipitate was washed with water and air dried to provide 380 mg of 5,6-difluoro-1H-benzo [d] imidazole (8).

Example 5: Synthesis of 5,6-Dimethoxy-1H

benzo [d] imidazola

MeO4 / NH2 HCOOH

MeO "^^ NH2 MW @ 220 ° C

9 10

The title compound 5,6-dimethoxy-1H

benzo [d] imidazole (10) was made by heating 4,5-dimethoxy-1,2-phenylenediamine dihydrochloride (9) in microwave formic acid at 220 ° C, followed by concentration in vacuo.

Example 6: Synthesis of 6-Fluoro-1H-

benzo [d] imidazole (11) and 6- (trifluoromethyl) -ΙΗ-benzo [d] imidazole (12)

The title compounds were made as described in US Patent Publication No. 2004/0087601, by some of the inventors of the present invention. Example 7: Benzimidazole (13), 5-Azabenzimidazole (14), 6-Chloro-5-fluorobenzimidazole (15) and 5-Methylbenzimidazole (16)

The title compounds were commercially

available.

13 14 15 U

Example 8: Synthesis of Primary Amine, Pyrazine-2

ilmethanamine

1. Raney NUH2

N 2 CN NH 3 / Me OH 2. Boc 2 O I CH 2 Cl 2 N chromatography J7 3. TFA / CH 2 Cl 2 18

A Raney nickel catalyst was carefully washed with THF and methanol, making sure that the catalyst remained wet. The weight of the wet catalyst was 2.5 g after washing. This material was added to a solution of pyrazinecarbonitrile (17) (3.0 g) in 7 N methanolic ammonium (120 mL). The mixture was stirred under a hydrogen atmosphere at 50 p.s.i. for 1.5 hours. The mixture was filtered and the filtrate was concentrated in vacuo to afford the crude title compound. Purification was performed by converting crude amine to tertiary butyl carbamate with excess of tertiary butyl dicarbonate in methylene chloride. Column chromatography (70: 27: 3 hexanes: ethyl acetate: methanol) provided 0.50 g of pure tertiary butyl pyrazine-2-ylmethylcarbamate. Pyrazine-2-

Pure NH 1 ilmethanamine (18) was obtained as the TFA salt from carbamate deprotection with 1: 1 TFA / CH 2 Cl 2.

Example 9: Synthesis of 3-Aminomethyl-2-

fluoropyridine

CN

CC

19

H2, Pd / C

BOH

20

A round bottom flask was charged with 0.3

g (2.46 mM) 3-cyano-2-fluoropyridine (19), which was then diluted with 20 mL EtOH. The solution was heated with argon and then while under an argon blanket 60 mg of 10% Pd / C (20% by weight) was added. The system was then sealed for 10 septum and placed under vacuum. A hydrogen flask was then added and the reaction was stirred for three hours (followed by TLC). The reaction was put under vacuum again, then exposed to air and filtered (keeping the catalyst moist). The resulting solution was dried and evaporated to give 0.28 g (90%) of the title compound, 15-aminomethyl-2-fluoropyridine (20).

Example 10: Synthesis of 3-Aminomethyl-6-methoxypyridine (21), 3-Aminomethyl-6-methylpyridine (22) and 3-Aminomethylquinoline (23)

The title amines were obtained from the corresponding nitriles using the same procedure as that used to obtain 3-aminomethyl-2-fluoropyridine (20) from 3-cyano-2-fluoropyridine (see Example 9).

OMe

21 22 23

Example 11: Synthesis of 3-Aminomethyl-2-Methoxy

pyridine NaOH3CN

CH4 NH4QAC

^ OMe 25

MeOH

24

A round bottom vial was loaded with 0.44

g (3.23 mM) 2-methoxy-3-pyridine carboxaldehyde (24), 1.24 g (16.15 mM) ammonium acetate and 0.61 g (19.69 mM) cyanoborohydride sodium The flask was then heated with argon and then 5 mL of dry MeOH was added via syringe. The reaction was stirred for 2 days and at this point the MeOH was evaporated. 25 mL of water was added and the mixture was left at pH = 2 with concentrated HCl. This was extracted twice with EtOAc to remove the alcohol side product. The mixture was left at pH = 10 using sodium hydroxide pellets, saturated with NaCl and extracted twice with DCM and once with EtOAc. The combined organics were dried and evaporated to give

0.31 g (69%) of 3-aminomethyl-2-methoxypyridine (25).

using the same procedure as that used to obtain 3-aminomethyl-2-methoxypyridine from carboxaldehyde of

2-methoxy-3-pyridine ((24), Example 11).

Example 12: Synthesis of 3- (Î ± -aminoethyl) -2-

chloropyridine (26)

26

The title amine was obtained from ketone.

20

Example 13: Synthesis of 3-Aminomethyl-4-

methylpyridine

27

28 10

A round bottom flask was charged with 0.45 g (3.30 mM) of 4-methylnicotinamide (27). The vial was heated with argon and 50 mL of dry THF was added via syringe. The resulting solution was cooled to 0 ° C and 2.5 mL (4.96 mM) of a 2 M solution of a borane-dimethylsulfide complex (in THF) was added. A bubbler was attached and the solution allowed to warm to RT overnight. The solution was cooled with MeOH, dried and evaporated to give 0.38 g (95%) of

3-Aminomethyl-4-methylpyridine (28).

Example 14: Hydronaphthalene-1-amine synthesis

of 5-fluoro-1,2,3,4-tetra-

THE

Me

29

1. 9-BBN / THF

2. 1 ~ Fluaro-2-iodobenzene, NaOMe1 Pd (Cippf) 2Cl2

MeO

NaOH

MeOH

1. (CICO) 2 cat DMF CH2Cl2

2. AICI3

HONH2-HCl NaOAc / EtOH

H2, Pd / C EtOH

of methyl

(2- (4-

4- (2-fluorophenyl) butanoate methylbutanoate) fluorobenzene, (30))

A round-bottom flask was sealed with an argon-heated rubber septum, then charged with 5.32 mL of methyl 3-buteneoate (29) and 100 mL of a 0.5 M solution of 9-BBN. in THF. The solution was stirred at RT for three hours. A 2 neck round bottom flask was equipped with a condenser and heated with argon, then charged with 7.35 g of sodium methoxide and 1.11 g of Pd (dppf) 2C12. To this mixture was added 20 mL of dry THF and 5.22 mL of 1-fluoro-2-iodobenzene. The hydroboration solution was added by cannula and the resulting mixture was refluxed for 16 hours. The solution was cooled to RT, diluted with 150 mL of water and extracted three times with ether. The combined organic layers were washed with brine, dried and evaporated. Column chromatography (5% EtOAc / hexanes) gave 1.79 g of methyl 4- (2-fluorophenyl) butanoate (30).

4- (2-Fluorophenyl) butanoic acid (31)

A round bottom flask was charged with 1.795 g of 2- (4-methylbutanoate) fluorobenzene, which was dissolved in 17 mL MeOH. To this solution was added a solution of 1 g of sodium hydroxide. The resulting mixture was stirred for 20 hours at RT. The solvent was evaporated and the crude material diluted with 15 mL of 0.5 M HCl. Extraction with DCM three times gave 1.17 g 10 (92%) of 4- (2-fluorophenyl) butanoic acid (31).

5-Fluoro-3,4-dihydronaphthalene-1 (2H) -one (32)

A round bottom flask was charged with 0.15 g of 4- (2-fluorophenyl) butanoic acid, which was dissolved in 20 mL of DCM and cooled to 0 ° C. Oxalyl chloride (0.15 mL) was added, followed by 1 drop of DMF. A drying tube was coupled and the solution was stirred at 0 ° C for two hours. Aluminum chloride (0.121 g) was added and the solution was allowed to slowly warm overnight at RT. The mixture was poured into frozen water and extracted three times with 20 DCM. The combined organic layers were washed with 0.5 M NaOH and brine. The organic phase was dried, evaporated and purified by column chromatography (eluting with 20% EtOAc / Hexanes) to give 0.07 g (53%) of 5-fluoro-3,4-dihydronaphthalene-1 ( 2H) -one (32).

5-fluoro-1,2,3,4-tetrahydronaphthalene-1-amine

(34)

A round bottom flask was charged with 0.5 g of 5-fluoro-3,4-dihydronaphthalene-1 (2H) -one, 0.28 g of hydroxylamine hydrochloride and 0.34 g of sodium acetate. A condenser was coupled and the flask was purged with argon. 20 mL of dry EtOH was added and the mixture was stirred at reflux for 18 hours. The solution was cooled to RT, diluted with EtOAc and washed with water. The organic phase was dried with sodium sulfate and evaporated to give 0.5 g of 5-fluoro-3,4-dihydronaphthalene-1 (2H) -one oxime intermediate (33), which was reduced with Pd / C in EtOH with hydrogen (50 psi) to give 0.43 g (86%) of 5-fluorofluoromethane.

1,2,3,4-tetrahydronaphthalene-1-amine (34). Example 15: Synthesis of 8-fluorochroman-4-amine

HO

3-Bromo propionic acid

NaOH / H2O

HO

35

1. (CICO) 2 cat. DMF CH2CI2

2. AICI3

HONH2-HCi

NaOAc / EtOH

H2l Raney Ni

EtOH

38

3- (2-Fluorophenoxy) propanoic acid (36)

A mixture of 2-fluorophenol (35) (15 g), 3-bromopropanoic acid (20 g) and NaOH (11 g) was refluxed in 50 mL of water. The solution was cooled to RT and acidified to pH = 2 with 3 M HCl. The resulting precipitate was isolated by filtration to obtain 9.27 g of the title compound as a white solid. The filtrate was extracted three times with EtOAc to obtain 2.5 g of the less pure compound (36).

8-fluorochroman-4-one (37)

Oxalyl chloride (8.79 mL) and a drop of DMF were added to a frozen solution of 3- (2-fluorophenoxy) propanoic acid (9.27 g) in DCM (50 mL). The solution was stirred at 0 ° C for two hours, aluminum chloride (7.39 g, 15.54 mM) was added and the solution was stirred for 16 hours at RT. The mixture was poured into frozen water and extracted three times with DCM. The combined organics were washed with 0.5 M NaOH and brine, dried, evaporated and purified by column chromatography (eluting with 20% EtOAc / Hex) to give 20 8-fluorochroman-4-one (37) ( 8.20 g, 98%).

8-fluorochroman-4-amine (39)

A round bottom flask was charged with 8-fluorochroman-4-one (8.2 g), hydroxylamine hydrochloride (3.78 g) and sodium acetate (4.46 g). A reflux condenser was added, the flask was purged with argon, dry EtOH (20 mL) was added and the mixture was stirred at reflux for 18 hours. The solution was cooled to RT, diluted with EtOAc and washed with water. The organic phase was dried and evaporated to give intermediate 8-fluorochroman-4-one oxime (38), which was reduced.

with Raney Nickel in 50 psi EtOH to yield the title amine (39) (4.69 g, 57%).

(3.40 g), methyl 2-methoxy acetate (2.44 g) and Novozyme 435 (Aldrich, 0.68 g) in anhydrous methyl tertiary butyl ether (75 mL) was heated to reflux under argon for two hours (at this time the ratio of acylated to nonacylated product was 1: 1 by HPLC - high pressure liquid chromatography). The solid that formed upon cooling was collected by filtration and dissolved in EtOAc. The mixture was filtered to remove the biocatalyst and washed once with 0.5 M HCl to remove any remaining (S) -amine. The solvent was evaporated and the product was recrystallized from tert-butyl methyl ether to obtain (R) -N- (8-fluorochroman-4-yl) -2-methoxy acetamide (0.78 g). The reaction solvent and the recrystallization "mother liquor" were washed three times with 0.5 M HCl and concentrated to obtain (R) -N- (8-fluorochromanon).

Additional 4-yl) -2-methoxy acetamide (0.83 g). The combined acidic aqueous layers were made basic by NaOH and extracted with

F

8-fluorochroman-4-amine resolution

F

Novozyme 435

F

MeOCH2COOMe

t-BuOMe

F

NH2

39b

Briefly, a mixture of 8-fluorochroman-4-amine DCM to obtain (5) -8-fluorochroman-4-amine (39a) (1.6 g). A solution of (R) -N- (8-fluorochroman-4-yl) -2-methoxyacetamide (0.78 g) in 8 M HCl in EtOH (50 mL) was heated at reflux for four hours. The solvents were removed from the cooled reaction mixture, the resulting solid was taken up in 50 mL of 0.5 M NaOH, salted with NaCl (s) and extracted four times with DCM to obtain (R) -8-fluorochroman-4. -amine (0.48 g (87%)) (39b). The% ee was conferred by chiral HPLC method: Chiralcel OD-II (0.46 x 25 cm analytical column, Daicel Chemical Industries);

isocratic, 5% (0.05% TFA / EtOH) and 95% (0.05% TFA / Hex), R t = 7.2 min (S) enantiomer, R t = 9.2 (R) enantiomer .

Example 16: Synthesis of 2- (1H-Benzo [d] imidazola-1-yl) -9- ((R) -8-fluorochromano-4-yl) -7H-purine-8 (9H) -one

2- (1H-Benzo [d] imidazol-1-yl) -N - ((R) -8-fluorochroman-4-yl) -5-nitropyrimidin-4-amine

(R) -8-fluorochroman-4-amine (60 mg, Example 15) was added to a solution of 2,4-dichloro-5-nitropyrimidine (70 mg) and DIEA (0.14 mL) in THF (5 mL) at -78 ° C. The reaction mixture was stirred for an additional 15 min at -78 ° C, then removed from the cold bath and warmed to RT. A molar solution of benzimidazole sodium salt (0.7 ml, shelf-life solution prepared by the addition of sodium hydride to a benzimidazole solution in THF) was added to the reaction intermediate ((R) -2-chloroacetate). N- (8-fluorochroman-4-yl) -5-nitropyrimidin-4-amine)

and the resulting mixture was stirred overnight at RT. Purification by column chromatography (elution with MeOH / DCM) gave the title compound (120 mg). MH + = 407.

2- (1H-Benzo [d] imidazol-1-yl) -9 - ((R) -8-fluorochromano-4-yl) -7H-purine-8 (9H) -one

A solution of sodium hydrosulfite (Tec., 0.5 g)

and freshly prepared sodium bicarbonate (0.25 g) in H 2 O (5 mL) was added to a solution of the above nitro compound (120 mg) in THF (10 mL). The mixture was stirred vigorously for 30 min, then extracted with EtOAc (2 x) and DCM (2 x), the combined organics were washed with brine, dried, filtered and concentrated to obtain 2- (ΙΗ-benzo intermediate). [d] imidazol-1-yl) -N4 - ((R) -8-fluorochroman-4-yl) pyrimidin-4,5-diamine, which was used as such in the next step.

Carbonyldiimidazole (0.2 g) was added to a solution of the above amine in THF (10 mL). The resulting mixture was stirred overnight at RT, silica gel was added, the solvents were then removed under reduced pressure and purified by column chromatography, eluting with 5% MeOH / DCM to obtain the title product (28 mg), MH + = 403, 1H NMR (CDCl3) δ 10.6 (s, 1H), 8.9 (s, 1H), 8.3 (s, 1H), 7.8 (m , 2H), 7.3 (m, 2H), 7.0 (m, 1H), 6.7 (m, 1H), 5.9 (dd, 1H), 4.7 (m, 1H), 4 , 4 (t, 1H), 2.7 (m, 1H), 2.3 (m, 1H) ppm, 19 F NMR δ-135.7 (m). Chiral CLAP - no evidence of another enantiomer. Method: Chiralcel OD-II (0.46 x 25 cm analytical column, Daicel Chemical Industries), isocratic, 15% (0.05% TFA / EtOH) and 85% (0.05% TFA / Hex), R t = 19.5 min enantiomer- (R), R t = 22.4 min enantiomer- (S).

Example 17: Non-regiospecific synthesis of benzimidazole purinone derivatives: Synthesis of 5-Nitro-N- (pyridin-3-ylmethyl) -2- (6- (trifluoromethoxy) -β-benzo [d] imidazole

1-yl) pyrimidin-4-amine (42) and 5-nitro-N- (pyridin-3-ylmethyl) -2- (5- (trifluoromethoxy) -β-benzo [d] imidazol-1-yl) pyrimidine -4-amine

(44) 41

43

F3C

F3CO H

45

THE

NO2

44

2-Chloro-5-nitro-N- (pyridin-3-ylmethyl) pyrimidin-4-amine (41)

A solution of 2,4-dichloro-5-nitropyridine (40) (5 g) in methylene chloride (60 mL) was cooled to -78 ° C and treated with 3- (aminomethyl) pyridine (2.8 g). The mixture was stirred at -78 ° C for six hours and concentrated in vacuo at RT to provide 2-chloro-5-nitro-N- (pyridin-3-ylmethyl) pyrimidin-4-amine

(41) crude, which was used without further purification.

5-Nitro-N- (pyridin-3-ylmethyl) -2- (6- (trifluoromethoxy) -β-benzo [d] imidazola-1-yl) pyrimidin-4-amine

(42) and 5-nitro-N- (pyridin-3-ylmethyl) -2- (5- (trifluoromethoxy) -1H-benzo [d] imidazol-1-yl) pyrimidin-4-amine (44)

crude acetimiditrile-4-amine (52 mg) in acetonitrile (10 mL) was treated with 6- (trifluoromethoxy) -ΙΗ-benzo [d] imidazole (40 mg), potassium carbonate (0.5 g) and heated at 80 ° C for four hours. The mixture was diluted with water and extracted with methylene chloride. The organic layer was separated, dried over sodium sulfate, filtered and concentrated in vacuo. Column chromatography (70: 22: 8 methylene chloride: ethyl acetate: methanol) provided 12 mg of 5-nitro-N- (pyridin-3-ylmethyl) -2- (5- (trifluoromethoxy) -4- benzo [d] imidazol-1-yl) pyrimidin-4-amine as the first

A suspension of 2-chloro-5-nitro-N- (pyridine-3-isomer elution) and 15 mg of 5-nitro-N- (pyridin-3-ylmethyl) -2- (6- (trifluoromethoxy) -β- benzo [d] imidazol-1-yl) pyrimidin-4-amine as the second elution isomer.

High Rf Isomer: 1 H-NMR (CDCl 3) δ 9.2 (s, 1H), 8.9 (s, 1H), 8.8 (m, 1H), 8.6 (s, 1H), 8.5 (d, 1H), 8.2 (d, 1H, determined: benzimidazole ring H-7), 7.6 (d, 1H), 7.6 (s, 1H, determined: H-4 of benzimidazole ring), 7.2 (dd, 1H), 4.9 (d, 2H).

Low Rf isomer: 1H-NMR (CDCl3) δ 9.2 (s, 1H),

8.9 (s, 1H), 8.8 (m, 1H), 8.6 (s, 1H), 8.5 (d, 1H), 8.2 (s, 1H), ring H-7 benzimidazole), 7.7 (d, 1H, found: H-4

benzimidazole ring), 7.6 (d, 1H), 7.2 (dd, 1H), 7.1 (d, 1H),

4.9 (d, 2H).

Example 18: Non-regiospecific synthesis of benzimidazole purinone derivatives: Synthesis of 9- (Pyridin-3-ylmethyl) -2- (6- (trifluoromethoxy) -β-benzo [d] imidazol-1-yl) -7H-

purine-8 (9H) -one (43) and 9- (pyridin-3-ylmethyl) -2- (5-

(trifluoromethoxy) -β-benzo [d] imidazol-1-yl) -7H-purine-8 (9H) -one

(45)

The title compounds were synthesized from 5-nitro-N- (pyridin-3-ylmethyl) -2- (6- (trifluoromethoxy) -1H-

benzo [d] imidazol-1-yl) pyrimidin-4-amine (42) and 5-nitro-N- (pyridin-3-ylmethyl) -2- (5- (trifluoromethoxy) -β-benzo [d] imidazol-4-yl)

1-yl) pyrimidin-4-amine (44) using the same procedures as for converting (R) -tertiary butyl 2,4-

(R) -Tertiary-butyl (2-oxo-3- (1-dimethyl) -benzyl (5-nitro-6- (1- (pyridin-3-yl) ethylamino) pyridin-2-yl) carbamate (pyridin-3-yl) ethyl) -2,3-dihydro-1H-imidazo [4,5-b] pyridin-5-yl) carbamate ((67); Example 22 below).

6-Trifluoromethoxy (non-salt) isomer: 1 H-NMR (CD 3 OD) δ 9.3 (s, 1H), 8.8 (br s, 1H), 8.6 (s, 1H, determin: H-7 benzimidazole ring), 8.6 (m, 1H), 8.4 (s, 1H), 8.1 (d, 1H), 7.9 (d, 1H, benzimidazole), 7.5 (dd, 1H), 7.4 (dd, 1H), 5.4 (s, 2H).

5-Trifluoromethoxy (non-salt) isomer: 1 H-NMR (CD 3 OD) δ 9.3 (s, 1H), 8.8 (s, 1H), 8.7 (d, 1H, determin: H-7 of benzimidazole ring), 8.5 (d, 1H), 8.3 (s, 1H), 7.9 (d, 1H), 7.6 (s, 1Η, determ .: benzimidazole ring H-4 ), 7.4 (dd, 1H), 7.3 (dd, 1H), 5.3 (s, 2H).

Example 19: Non-regiospecific synthesis of an oxo-imidazopyridine and an imidazopyridine derivative:

5- (1H-Benzo [d] imidazol-1-yl) -3- (pyridin-3-ylmethyl) -1H-imidazo [4,5-b] pyridin-2 (3H) -one (50)

CL ·

N == y

I I ^

X - 9 DMSCWH2O

49

6-1H-Benzo [d] imidazol-1-yl) -S-nitro-N- (pyridine)

3-ylmethyl) pyridine-1-amine (48)

A solution of 2,6-dichloro-3-nitropyridine (46) (0.5 g) in acetonitrile (20 mL) was cooled to 0 ° C and treated with triethylamine (0.36 mL), followed by 3- (aminomethyl) pyridine (0.26 mL). The mixture was stirred for 30 minutes at 0 ° C and eight hours at RT. The resulting solution, which contained the 6-chloro-

3-nitro-N- (pyridin-3-ylmethyl) pyridine-2-amine (47) was

Transferred to a sealed tube containing benzimidazole (0.84 g) and potassium carbonate (3 g) and heated at 70 ° C for 16 h. The mixture was cooled and filtered. The precipitates were washed with water and air dried to provide 239 mg of the title compound (48).

5- (1H-Benzo [d] imidazol-1-yl) -3- (pyridin-3-ylmethyl) -1H-imidazo [4,5-b] pyridin-2 (3H) -one (50) and 5- (1H-benzo [d] imidazol-1-yl) -3- (pyridin-3-ylmethyl) -3H-imidazo [4,5-

b] pyridine (51)

A solution of 6- (β-benzo [d] imidazol-1-yl) -3-nitro-N- (pyridin-3-ylmethyl) pyridine-2-amine (48) (50 mg) in 1 mL 10 of DMSO was treated with a solution of Na 2 S 2 O 4 (300 mg) in 1 mL of water. The mixture was stirred for two hours and diluted with 50 mL of ethyl acetate. The mixture was washed three times with 50 mL aliquots of a saturated sodium chloride solution, dried over sodium sulfate, filtered and concentrated in vacuo to afford intermediate 6- (β-benzo [d] imidazol-1-yl). ) -N2- (pyridine-3-

ylmethyl) pyridine-2,3-diamine (49). Half of the intermediate was dissolved in methylene chloride (2 mL) and treated with 1,1'-carbonyldiimidazole (46 mg) at RT for 16 h. The resulting crude mixture was purified by preparative TLC (1,000 micron, 5% MeOH / CH 2 Cl 2) to provide 7.1 mg of 5- (ΙΗ-benzo [d] imidazol-1-yl) - 3- (pyridin-3-ylmethyl) -1H-

imidazo [4,5-b] pyridine-2 (3H) -one (50): 1 H-NMR (CDCl 3) δ 10.0 (br s, 1H), 8.9 (s, 1H), 8.6 ( d, 1H), 8.5 (s, 1H), 7.9 (m, 2H), 7.8 (m, 1H), 7.6 (d, 1H), 7.4 (m, 2H), 7.4 (m, 1H), 7.3 (d, 1H), 5.2 (s, 2H).

Example 20: Regiospecific Synthesis: Synthesis of

3- (9- (2,6-Difluorobenzyl) -8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-5-carbonitrile H2N

avVcl

MeO

OMe

ASS

DlEA / THF

H 2 J 2 H 2 N 2 XJI

NO2

DIEA / THF

1 ΝγγΚ.

OMe N ^ Anhz F 54

THF

Η

Niió = ° 55

TFA / Et3SiH

OMe

Bocp

TEA

MeOH

CH3CN

DCM

= * ■

JCC1

NC - ^^

NO2

F

NtH / DMF

56

NJO2 μ

. 'Ν-τΓΝ ^ τΛν_

1 Ν ^ Λ- ^ 0 THF 'H2O

Mouth

Mouth

57

NH2

(MeO) 3CH

Ηϊ ^ 0 = ° PTsOH

MeOH

CN if

CN 59

Mouth

N2- (2,4-Dimethoxybenzyl) -N4- (2,6-difluorobenzyl) -5-nitropyrimidine-2,4-diamine (53)

2,6-Difluorobenzylamine (0.24 mL) was added by dripping over a minute to a solution of 2,4-dichloro-5-nitropyrimidine (40) (0.388 g) and DIEA (0.77 mL) in THF in a cold bath set to -78 ° C. The reaction mixture was stirred for an additional 15 min at -78 ° C, then removed from the cold bath and allowed to warm to RT. Additional DIEA (0.77 5 mL) was added to the reaction intermediate (N- (2,6-difluorobenzyl) -2-chloro-5-nitropyrimidine-4-amine) (52) followed by the addition of 2,4-dimethoxybenzylamine (0.30 mL) and the resulting mixture was stirred overnight at RT. Purification by column chromatography (eluted with 1 and 2.5% MeOH / DCM) gave 10 N2- (2,4-dimethoxybenzyl) -N4- (2,6-difluorobenzyl) -5-nitropyrimidine-2,4 diamine (53) (0.80 g), MH + = 432.

2- (2,4-Dimethoxybenzylamino) -9- (2,6-difluorobenzyl) -7H-purine-8 (9H) -one (55)

Raney NI was added to a solution of N2- (2,4-dimethoxybenzyl) -N4- (2,6-difluorobenzyl) -5-nitropyrimidine-

2,4-diamine (0.80 g) in THF (50 mL) under argon heating. The suspension was evacuated, charged with hydrogen (flask) and stirred for 16 h. The resulting mixture was filtered through a celite plug, which was thoroughly rinsed with THF and MeOH, to obtain 20 N2- (2,4-dimethoxybenzyl) -N4- (2,6-difluorobenzyl) pyrimidine-2,4,5 - triamine (54), which was used as such in the next reaction.

Carbonyldiimidazole (0.93 g) was added to a solution of N2- (2,4-dimethoxybenzyl) -N4- (2,6-

difluorobenzyl) pyrimidine-2,4,5-triamine (54) in THF (20 mL) and the resulting mixture was stirred overnight at RT; The solvents were then removed under reduced pressure and taken up in EtOAc and washed three times with water. The organics were dried, filtered, evaporated and purified by column chromatography, eluting with 2.5 and 4% MeOH / DCM to give 2- (2,4- 30 Dimethoxybenzylamine) -9- (2,6-difluorobenzyl) -7H-purine-8 (9H) -one (55) (0.58 g), MH + = 428.

Tertiary Butyl 9- (2,6-difluorobenzyl) -2-amino-8-oxo-8,9-dihydropurine-7-carboxylate (57)

A 1: 1 solution of TFA / DCM (10 mL) was added to 2- (2,4-dimethoxybenzylamine) -9- (2,6-

difluorobenzyl) -7H-purine-8 (9H) -one (55) (0.58 g) and stirred for 30 min, after which triethyl silane (2 mL) was added and the mixture was stirred for an additional 4 h. The solvents were removed under vacuum, the residue taken up in minimum MeOH and triturated with Et 2 O to obtain 9- (2,6-difluorobenzyl) -2-amino-7H-purine-8 (9H) -one TFA salt (56) (0.55 g). MH + = 278, as a salmon colored solid.

9- (2,6-Difluorobenzyl) -2-amino-7H-purine-8 (9H) -one (0.55 g) was dissolved in a mixture of MeOH / ACN / DCM (40 mL), Et 3 N (2 mL). ) and di-tertiary butyl dicarbonate (0.61 g) were added and the mixture was stirred overnight at RT. Reaction solvents were removed and crude material was taken up in DCM and washed with H 2 O, evaporated and purified by column chromatography, eluting with 2 and 3% MeOH / DCM to give the title product (57) (0, 36 g). MH + = 378, MH + -BoC = 278 (major), (M + Na) + = 400 and (2M + Na) + = 777 were also observed.

Tertiary Butyl 9- (2,6-difluorobenzyl) -2- (5-cyano-

2-nitrophenylamino) -8-oxo-8,9-dihydropurine-7-carboxylate (58)

Sodium hydride (88 mg, 95%) was added, under argon heating, to a 9- (2,6-difluorobenzyl) -2-amino-8-oxo-8,9-dihydropurine solution of tertiary butyl -7-carboxylate

(57) (191 mg) and 3-fluoro-4-nitrobenzonitrile (415 mg) in DMF (5 mL) at -40 ° C. The reaction mixture was allowed to warm to -20 ° C for 3 h, then was cooled by the addition of heated saturated NH 4 Cl and once at RT the mixture was diluted with 25 EtOAc and separated. The organics were washed with brine (3x), dried, filtered and evaporated, purified by column chromatography (eluting with DCM and 1 and 2.5% MeOH / DCM) to obtain tertiary Butyl 9- (2.6 -difluorobenzyl) -2- (5-cyano-2-nitrophenylamino) -8-oxo-8,9-dihydropurine-7-carboxylate (58) (288 mg), MH + = 524.

Tertiary Butyl 9- (2,6-difluorobenzyl) -2- (6-cyano-

Β-benzo [d] imidazol-1-yl) -8-oxo-8,9-dihydropurine-7-carboxylate (60)

A freshly prepared solution of sodium hydrosulphite (tec. 1 g) and sodium bicarbonate (0.5 g) in H 2 O (10 mL) was added to a solution of the above nitro compound (58) (288 mg ) in THF (10 mL). The mixture was stirred vigorously for 5 min, extracted with DCM (3x), the combined organics were washed with brine, dried, filtered and concentrated to obtain tertiary butyl 9- (2,6-difluorobenzyl) -2- ( 2-amino-5-cyanophenylamino) -8-oxo-8,9-dihydropurine-7-carboxylate (59), which was used as such in the next step.

A catalytic amount of sodium monohydrate

para-toluene sulfonic acid was added to a solution of the above amine intermediate and trimethyl orthoformate (3 mL) in MeOH (10 mL). After 1 hr crude material was adsorbed onto silica gel and purified by column chromatography (eluting 10 with 1 and 2% MeOH / DCM) to obtain tertiary-Butyl 9- (2,6-difluorobenzyl) -2- ( 6-cyano-1H-benzo [d] imidazol-1-yl) -8-oxo-8,9-dihydropurine-7-carboxylate (60) (164 mg), MH + = 504 and MH + -BOC = 404.

3- (9- (2,6-Difluorobenzyl) -8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-5-carbonitrile (61)

A 1: 1 solution of TFA / DCM (10 mL) was added to Tertio Butyl 9- (2,6-difluorobenzyl) -2- (6-cyano-1-β-benzo [d] imidazol-1-yl) ) -8-oxo-8,9-dihydropurine-7-carboxylate (60) and stirred for 1 h. The solvents were removed in vacuo and the resulting solid was triturated with Et 2 O and suspended in 6 N HCl. Removal of solvents and trituration with Et 2 O from the resulting solid gave the title compound (61) (68 mg) as an HCl salt. MH + = 404.1 1H NMR (d6-DMS0) δ 11.8 (s, 1H), 9.2 (s, 1H), 8.8 (s, 1H), 8.0 (s, 1H), 7, 9 (d, 1H), 7.8 (broad s, 1H), 7.4 (d, 1H), 257.4 (quintet, 1H), 7.1 (m, 2H), 5.2 (s 2H) ppm, 19 F NMR δ -114.3 (m).

Example 21: Synthesis of 3- (8-Oxo-9- (tetrahydro-2H-pyran-4-yl) -8,9-dihydro-7H-purin-2-yl) -3H-benzo [d ] imidazole-5-carbonitrile (62).

The title compound can be synthesized using the same procedures as described for the synthesis of 3- (9- (2,6-difluorobenzyl) -8-oxo-8,9-dihydro-7H-purin-2-yl ) -3H-benzo [d] imidazole-5-carbonitrile ((61), Example 20).

1H NMR (d6-DMSO) 11.71 (s, 1H), 9.34 (s, 1H),

8.94 (d, J = 1.5 Hz, 1H), 8.38 (s, 1H), 8.00 (d, J = 8.1 Hz, 1H), 7.79 (dd, J = 8 , 1.5 Hz, 1H), 4.57 (m, 1H), 4.04 (m, 2H), 3.50 (m, 2H), 2.59 (m, 2H), 1.79 (m, 2H); Mass (MH +) 362.1.

Example 22: Regiospecific Synthesis of an Oxo-imidazopyridine Derivative: Synthesis of 3- (2-oxo-3 - ((R) -1- (pyridin-3-yl) ethyl) -2,3-dihydro-1H -imidazo [4,5-b] pyridin-5-yl) -3H-benzo [d] imidazole-5-carbonitrile CL X!

Tl

MeO

MeO

MeO

I JL TEA / THF

^ s * ^ xno2

46

Il H H

11 N ^ N_N

OMe

MeO

H1 Boc2OZDMAP CH2CI2

Il ^ 0c H 1 n ^ n ^ n

OMe

64.1

Meon ^ oc h o

Na2S2O4

1VS ^ I

65

CDI

THF / H2O / MeOH OMe

NH2

THF / 50 0C

66

• Q

Q

TFAZEt3SH

CH2Q2

N._

h2n ^ n ^ -r ^

Il I> = 0

67

H

68

H

Boc2O

K2CO3

CH3CN

.OMe

H

OyN ^ 1NyIvN _ „

'VS' l <b tea / thf

jÇF

MH / DMF

f »H V

çrO>

CN 70 koc

Na2S2O4

THF / H2O / MeOH

(MeO) 3CH

pTsOH

THF

TFAZCH2Cb

o>

N Boc

72

Nsv-N N

x ^ r

(R) -N6- (2,4-Dimethoxybenzyl) -3-nitro-N2- (1- (pyridin-3-yl) ethyl) pyridine-2,6-diamine (64)

A solution of 2,6-dichloro-5-nitropyridine (46) (0.5 g) in THF (20 mL) was cooled to 0 ° C and treated with 1.6 mL of triethylamine, followed by (R) -1- pyridin-3-yl-ethylamine (300 µL). The mixture was stirred for 1.5h, then warmed to RT and stirred for another 20h. 2,4-Dimethoxybenzylamine (0.8 mL) was added and the mixture was heated at 50 ° C for four hours. The mixture was diluted with ethyl acetate and washed twice with saturated sodium chloride solution. The organic layer was separated, dried over sodium sulfate, filtered and concentrated. Column chromatography (50-100% ethyl acetate in 5 hexanes) provided 761 mg of (R) -N6- (2,4-Dimethoxybenzyl) -3-nitro-N2- (1- (pyridine-3- yl) ethyl) pyridine-2,6-diamine (64).

(R) Tertiary Butyl 2,4-dimethoxybenzyl (5-nitro-6- (1- (pyridin-3-yl) ethylamino) pyridin-2-yl) carbamate (65)

A solution of (R) -N6- (2,4-dimethoxybenzyl) -3-10 nitro-N2- (1- (pyridin-3-yl) ethyl) pyridine-2,6-diamine (64) (367 mg) in methylene chloride (20 mL) was treated with di-tertiary butyl dicarbonate (1.0 g) and 4-dimethylaminopyridine (22 mg). The mixture was stirred for 16 h and concentrated in vacuo. Column chromatography (50 -> 100% ethyl acetate in 15 hexanes) provided 500 mg of (R) -tertium Butyl 2,4-dimethoxybenzyl (5-nitro-6- (1- (pyridin-3-yl)). ) ethylamino) pyridin-2-yl) carbamate

(65).

(R) Tertiary Butyl 2,4-dimethoxybenzyl (2-oxo-3- (1- (pyridin-3-yl) ethyl) -2,3-dihydro-1H-imidazo [4,5-b] pyridin-5-yl) carbamate (67)

A solution of (R) -Tertiary butyl 2,4-dimethoxybenzyl (5-nitro-6- (1- (pyridin-3-yl) ethylamino) pyridin-2-yl) carbamate (500 mg) in THF (25 mL) ) was treated with an aqueous solution comprising 2 g of Na 2 S 2 O 4 and 1 g of NaHCO 3 in 20 mL of water, followed by 1 mL of methanol. The mixture was stirred for 30 minutes, then diluted with ethyl acetate and washed with saturated sodium chloride solution. The organic layer was separated, dried over sodium sulfate, filtered and concentrated to afford intermediate (R) -Tertiary butyl 2,4-dimethoxybenzyl (5-30 amino-6- (1- (pyridin-3-yl) ethylamino) ) pyridin-2-yl) carbamate

(66). The intermediate was dissolved in THF (50 mL) and treated with

1,1'-carbonyldiimidazole (0.5 g) at 50 ° C for 20 h. The mixture was concentrated and purified by column chromatography (2-5% MeOH in methylene chloride) to afford 413 mg of (R) -.

Tertiary Butyl 2,4-dimethoxybenzyl (2-oxo-3- (1- (pyridine-3-

yl) ethyl) -2,3-dihydro-1H-imidazo [4,5-b] pyridin-5-yl) carbamate

(67). (R) -Tertiary Butyl 5-amino-2-oxo-3- (1- (pyridin-3-yl) ethyl) -2,3-dihydro-imidazo [4,5-b] pyridine-1-one carboxylate (69)

A solution of (R) -Tertiary butyl 2,4-dimethoxybenzyl (2-oxo-3- (1- (pyridin-3-yl) ethyl) -2,3-dihydro-1H-5-imidazo [4, 5-b] pyridin-5-yl) carbamate in methylene chloride (15 mL) was treated with TFA (15 mL) and triethyl silane (1.0 mL) for one hour. The mixture was concentrated to afford intermediate (R) -5-amino-3- (1- (pyridin-3-yl) ethyl) -1H-imidazo [455-b] pyridin-2 (3H) -one (68) , which was dissolved in acetonitrile (50 mL) and stirred vigorously with di-tertiary butyl dicarbonate (1.0 g) and potassium carbonate (3.0 g) for 2 h. Methylene chloride (200 mL) and water (100 mL) were added and the organic layer was separated. The aqueous layer was extracted with another 100 mL of methylene chloride. The combined organic layers were separated, dried over sodium sulfate, filtered and concentrated. Column chromatography (2 -> 3 -> 4% MeOH in methylene chloride) provided 235 mg of (R) tert-Butyl 5-amino-2-oxo-3- (1- (pyridin-3-yl) ) ethyl) -2,3-dihydroimidazo [4,5-b] pyridine-1-carboxylate (69).

(R) -Tertiary Butyl 5- (5-cyano-2-nitrophenylamino) -

2-OXO-3- (1- (pyridin-3-yl) ethyl) -2,3-dihydroimidazo [4,5-

b] pyridine-1-carboxylate (70)

A solution of (R) -Tertiary butyl 5-amino-2-oxo-3- (1- (pyridin-3-yl) ethyl) -2,3-dihydroimidazo [4,5-b] pyridine-1 - carboxylate (94 mg) and 3-fluoro-4-nitrobenzonitrile (225 mg) in DMF (6 mL) were cooled to -25 ° C and treated with NaH (60 wt% in mineral oil 75 mg) and allowed to warm slowly to -15 ° C. The mixture was stirred for four hours at -20 ° C to -15 ° C, then diluted with EtOAc and cooled with a saturated ammonium chloride solution. The organic phase was washed three times with brine, separated, dried over sodium sulfate, filtered and concentrated. Column chromatography (2% MeOH in methylene chloride) provided 100 mg of (R) -tertiary-Butyl 5- (5-cyano-2-nitrophenylamino) -2-oxo-3- (1-pyridine-3-one). yl) ethyl) -2,3-dihydroimidazo [4,5-b] pyridine-1-carboxylate (70). Tertiary Butyl 5- (6-cyano-1H-benzo [d] imidazola-1-yl) -2-OXO-3- ((R) -1- (pyridin-3-yl) ethyl) -2,3- dihydroimidazo [4,5-b] pyridine-1-carboxylate (72)

A solution of (R) -Tertiary butyl 5- (5-cyano-2-nitrophenylamino) -2-oxo-3- (1- (pyridin-3-yl) ethyl) -2,3-dihydroimidazo [4,5-b] pyridine-1-carboxylate (70) (100 mg) in THF (5 mL) was treated with an aqueous solution comprising 0.5 g Na2S2O4 and 0.25 g NaHCO3 in 5 mL water . The mixture rapidly changed from a red to a slightly yellowish color, indicating a reduction in the nitro group. The mixture was diluted with ethyl acetate and washed with a saturated sodium chloride solution. The organic layer was separated, dried over sodium sulfate, filtered and concentrated to afford intermediate (R) - tertiary butyl 5- (2-amino-5-cyanophenylamino) -2-oxo-3- (1-pyridine-3 -yla) ethyl) -2,3-dihydroimidazo [4,5-b] pyridine-1-carboxylate (71). The intermediate was dissolved in THF (5 mL), DMF (1 mL) and trimethylortoformate (2 mL). The mixture was treated with 10 mg p-toluenesulfonic acid and stirred for 20 h. The mixture was diluted with ethyl acetate and washed once with saturated sodium bicarbonate and twice with saturated NaCl solution. The organic layer was separated, dried over sodium sulfate, filtered and concentrated. Column chromatography (2% MeOH in methylene chloride) provided 57 mg of tertiary-Butyl 5- (6-cyano-1H-benzo [d] imidazola-1-yl) -2-oxo-3 - ((R ) -1- (pyridin-3-yl) ethyl) -2,3-dihydroimidazo [4,5-b] pyridine-1-carboxylate (72).

3- (2-oxo-3 - ((R) -1- (pyridin-3-yl) ethyl) -2,3-dihydro-1H-imidazo [4,5-b] pyridin-5-yl) -3H-benzo [d] imidazole-5-carbonitrile (73)

A solution of tertiary butyl 5- (6-cyano-1H-benzo [d] imidazola-1-yl) -2-oxo-3 - ((R) -1- (pyridin-3-yl) ethyl)

2,3-Dihydroimidazo [4,5-b] pyridine-1-carboxylate (72) (57 mg) in methylene chloride (1 mL) was treated with TFA (1 mL) for one hour. The mixture was concentrated and the resulting TFA salt was converted to the HCl salt by dissolving in 5 mL of EtOH and adding 0.5 mL of concentrated HCl and then concentrating the solution in vacuo. The process was repeated and the resulting residue was dissolved in a minimum amount of methanol and triturated with 10

the addition of ethyl ether. After 3 grindings, 3- HCl salt

(2-oxo-3 ((R) -1- (pyridin-3-yl) ethyl) -2,3-dihydro-1 H-

imidazo [4,5-b] pyridin-5-yl) -3H-benzo [d] imidazola-5-carbonitrile

(73) (39 mg) was isolated as a light brown solid:

iH-

NMR (CD 3 OD) δ 9.9 (br s, 1H), 9.2 (s, 1H), 9.0 (m, 2H), 8.5 (s, 1H), 8.3 (m, 1H) , 8.2 (m, 1H), 8.1 (d, 1H), 7.9 (d, 1H), 7.8 (d, 1H), 6.3 (q, 1H), 2.3 ( d, 3H).

Example 24: Synthesis of 2- (1H-Benzo [d] imidazola-1-yl) -9- (cis-3-methyl-tetrahydro-2H-pyran-4-yl) -7H-purine-8 (9H ) - one

cW DIEA

THF

CI

Benzyrt.

K2CO3

THF

NO2

1. Ns2S2OiJ NaHCO3

THF / H2O / MeOH 2. CDi / THF

2-Chloro-N- (cis-3-methyl-tetrahydro-2H-pyran-4-yl) -5-nitropyrimidine-4-amine

To a suspension of 0.24 g of cis-3-methyl-tetrahydro-2H-pyran-4-amine hydrochloride salt (WO 2004/041161) and DIEA (1.5 mL) in THF (10 mL) at -78 ° C, 2,4-dichloromethane was added.

5-nitropyrimidine (0.72 g). The mixture was allowed to slowly reach room temperature and stirred for 16 hours. The mixture was diluted with EtOAc and washed 3 times with brine. The organic layer was separated, dried over sodium sulfate and concentrated in vacuo. Column chromatography (20-40% EtOAc / hexanes) provided 289 mg of the title compound.

2- (1H-Benzo [d] imidazola-1-yl) -N- (cis-3-methyl-tetrahydro-2H-pyran-4-yl) -5-nitropyrimidin-4-amine

To a solution of 2-chloro-N- (cis-3-methyl-tetrahydro-2H-pyran-4-yl) -5-nitropyrimidine-4-amine (115 mg) in acetonitrile (5 mL) was added carbonate. potassium (300 mg) and benzimidazole (150 mg). The mixture was stirred at 70 ° C for 2.5 hours. After diluting with 70 mL EtOAc, the mixture was washed with brine, dried over sodium sulfate and concentrated in vacuo. Column chromatography (50-100% EtOAc / hexanes) provided 99 mg of the title compound.

2- (1H-Benzo [d] imidazola-1-yl) -9- (cis-3-methyl-tetrahydro-2H-pyran-4-yl) -7H-purine-8 (9H) -one

To a solution of 2- (β-benzo [d] imidazola-1-yl) -N- (cis-3-methyl-tetrahydro-2H-pyran-4-yl) -5-nitropyrimidin-4-one

amine (51 mg) in THF (10 mL) was added a solution of sodium hydrosulphite (300 mg) and sodium bicarbonate (150 mg) in water (10 mL). The mixture turned blue briefly and then became colorless. Methanol (1 mL) was added to maintain homogeneity of the solution. The mixture was diluted with 70 mL EtOAc and washed twice with brine. The aqueous washes were extracted with other

50 mL of EtOAc and then the combined organic layers were dried over sodium sulfate and concentrated in vacuo to provide

2- (β-benzo [d] imidazola-1-yl) -N4- (cis-3-methyl-tetrahydro-2H-pyran-4-yl) pyrimidin-4,5-diamine. The diamine intermediate was dissolved in THF (5 mL) and treated with 1,1'-carbonyldiimidazole (80 mg) at 50 ° C for 16 hours. The mixture was diluted with 50 mL EtOAc and washed 3 times with brine. The organic layer was separated, dried over sodium sulfate and concentrated in vacuo. Column chromatography (2-4% MeOH / DCM) provided 19.3 mg of the title compound. 1H-NMR (300 MHz, 5%

of O at Γ0 OO 9 (s, IH), 8.5 (d, IH), 8.2 (s, IH), OO (d, D I --- I to r- COOQDO IH), 7, 4 (t, 1H), 7.3 (t, 1H), 4.7 (m, 1H), 4.2 (d (br), 1H), 3.9 (d, 1H), 3.7 ( d, 1H), 3.5 (m, 2H), 2.3 (t (br), 1H), 1.8 (d (br), 1H), 1.2 (d, 3H). Example 25 2- (5,6-Dichloro-1H-benzo [d] imidazola-1-yl) -9- ((R) -8-fluorochroman-4-yl) -7H-purine-8 (9H) -one

A solution of (R) -2-chloro-N- (8-fluorochroman-4-yl) -5-nitropyrimidine-4-amine in acetonitrile was treated with 5,6-dichlorobenzimidazole and potassium carbonate. The mixture was stirred at reflux for 6 hours, cooled to room temperature, diluted with 150 mL EtOAc and washed twice with 30 mL portions of water. The organic layer was separated, dried with magnesium sulfate, filtered and concentrated in vacuo. Purification by column chromatography (2% MeOH / DCM) gave intermediate nitropyrimidinamine. The title compound was synthesized from intermediate nitropyrimidinamine by the procedures described in Example 24. 1H-NMR (300 MHz, CDCl3) δ 8.7 (s, 1H), 8.3 (s, 1H), 8.2 (s, 1H), 7.8 (t, 1H), 7.0 (t, 1H), 6.6 (m, 2H), 5.9 (t, 1H), 4.6 (m, 1H) ), 4.4 (m, 1H), 3.0 (m, 1H),

2.3 (m, 1H).

2- (5,6-Dimethyl-1H-benzo [d] imidazola-1-yl) -9- ((R) -8-fluorochromano-4-yl) -7H-purine-8 (9H) -one

The title compound was synthesized from 5,6-dichlorobenzimidazole by the procedures described in Example 25. 1H-NMR (300 MHz, CDCl3) δ 9.6 (s, 1H), 8.8 (s, 1H), 8.3 (s, 1H), 8.0 (s, 1H), 7.6 (s, 1H), 7.0 (t, 1H), 6.7 (m, 1H), 5.9 (t , 1H), 4.7 (m, 1H), 4.5 (m, 1H), 3.2 (m, 2H), 2.4 (d, 6H). . 53 N

Çr

F3C

FaC '

ίο

15

53

9 - ((R) -8-Fluorochromano-4-yl) -2- (6- (trifluoromethyl) -β-benzo [d] imidazol-1-yl) -7H-purine-8 (9H) -one and 9 - ((R) -8-fluorochroman-4-yl) -2- (5- (trifluoromethyl) -1H-benzo [d] imidazol-1-yl) -7H-purine-8 (9H) -one

The title compound was synthesized from 5-trifluoromethylbenzimidazole (US 2 004/0087 601) by the procedures described in Example 25. Purification by column chromatography (2% MeOH / DCM) eluted the 6-trifluoromethyl isomer. first (1H-NMR (300 MHz, CDCl3) δ 8.8 (d, 2H), 8.4 (s, 1H), 7.9 (d, 1H), 7.6 (d, 2H), 7, O (t, 1H), 6.7 (m, 2H), 5.9 (t, 1H), 4.7 (m, 1H), 4.4 (m, 1H), 3.0 (m, 1H) ), 2.4 (m, 1H)), followed by 5-trifluoromethyl isomer (1H-NMR (300 MHz, CDCl3) δ 9.0 (s, 1H), 8.4 (s, 1H), 8 0.1 (s, 1H), 8.0 (d, 1H), 7.6 (d, 2H), 7.0 (m, 1H), 6.8 (m, 2H), 5.9 (t, H), 4.7 (m, 1H), 4.4 (m, 1H), 2.9 (m, 1H), 2.4 (m, 1H)).

N

&

N

20

N - ((R) -8-Fluorochromano-4-yl) -2- (3H-imidazo [4,5-c] pyridin-3-yl) -5-nitropyrimidin-4-amine and N - ((R) -8-

fluorochroman-4-yl) -2- (1H-imidazo [4,5-c] pyridin-1-yl) -5-nitropyrimidin-4-amine 10

15

20

The title compound was synthesized from 5-azabenzimidazole by the procedure described in Example 25. Purification by column chromatography (1% MeOH / DCM) provided N- ((R) -8-fluorochroman-4-yl ) -2- (3H-imidazo [4,5-c] pyridine-2

3-yl) -5-nitropyrimidin-4-amine as the first eluting isomer: (1H-NMR (300 MHz, CDCl3) δ 9.8 (s, 1H), 9.4 (s, 1H), 9, 2 (s, 1H), 8.9 (d, 1H), 8.6 (d, 1H), 7.8 (d, 1H), 7.1 (m, 2H), 6.9 (m, 1H) ), 5.8 (q, 1H), 4.6 (m, 1H), 4.4 (m, 1H), 2.6 (m, 1H), 2.4 (m, 1H)). N - ((R) -8-Fluorochroman-4-yl) -2- (1H-imidazo [4,5-

c] pyridin-1-yl) -5-nitropyrimidin-4-amine eluted second: (1H-NMR (300 MHz, CDCl 3) δ 9.4 (s, 1H), 9.2 (s, 1H), 9.1 (s, 1H), 8.9 (d, 1H), 8.6 (d, 1H), 8.4 (d, 1H), 7.1 (m, 2H), 6.9 (m , 1H), 5.7 (q, 1H), 4.5 (m, 1H), 4.4 (m, 1H), 2.6 (m, 1H), 2.4 (m, 1H)).

ΧΛ- °

H

9- ((R) -8-fluorochroman-4-yl) -2- (3H-imidazo [4,5-

c] pyridin-3-yl) -7H-purine-8 (9H) -one

The title compound was synthesized from N- ((R) -8-fluorochroman-4-yl) -2- (3H-imidazo [4,5-c] pyridin-3-yl) -5-nitropyrimidin-4 -amine by the procedures described in Example 24. 1H-NMR (300 MHz, CDCl3) δ 9.8 (s, 1H), 9.4 (s, 1H), 8.6 (d, 1H), 8.3 (m, 2H), 7.0 (t, 1H), 6.7 (m, 2H), 5.9 (t, 1H), 4.6 (m, 1H), 4.4 (m, 1H) 2.8 (m, 1H), 2.4 (m, 1H).

Nv ^ N μ

H

9- ((R) -8-fluorochroman-4-yl) -2- (1H-imidazo [4,5-

c] pyridin-1-yl) -purine-8 (9H) -one The title compound was synthesized from N- ((R) -8-fluorochroman-4-yl) -2- (1H-imidazo [4 , 5-c] pyridin-1-yl) -5-nitropyrimidin-4-amine by the procedures described in Example

g) in THF (100 mL) was treated with DIEA (6.3 mL) and 2,4-dimethoxybenzylamine (5.0 mL) and then stirred for 24 h. The solvent was evaporated and the crude mixture was dissolved in EtOAc 15 (100 mL). The solution was washed once with 1 M HCl and twice with saturated aqueous NaCl (100 mL each). The organic layer was separated, dried over Na 2 SO 4, filtered and concentrated in vacuo. Column chromatography (20% EtOAc / DCM) provided 9.25 g of the title compound.

4- (2,4-dimethoxybenzylamine) -3-aminobenzonitrile

nitrobenzonitrile (4.54 g) in THF (400 mL) was treated with a solution of sodium hydrosulphite (20 g) and sodium bicarbonate (10 g) in distilled water (350 mL). Enough methanol was immediately added (50 mL). mL) to keep the solution homogeneous. After 15 minutes, EtOAc (500 mL) and saturated aqueous NaCl (500 mL) were added and the organic layer was separated. The aqueous layer was extracted again with 400 mL of EtOAc. The organic layers

24. 1H-NMR (300 MHz, CDCl3) δ 9.3 (d, 2H), 8.4 (d, 1H), 8.3 (d,

2H), 7.0 (t, 1H), 6.7 (m, 2H), 5.9 (t, 1H), 4.6 (m, 1H), 4.4 (m, 1H), 2, 8 (m, 1H), 2.4 (m, 1H).

Example 26: Synthesis of 3- (9 - ((R) -6,8-

difluorochromano-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-

benzo [d] imidazole-5-carbonitrile

OMe

.OMe

OMe

DIEA

THF

10

4- (2,4-dimethoxybenzylamine) -3-nitrobenzonitrile

A solution of 4-fluoro-3-nitrobenzonitrile (5.0

A solution of combined 4- (2,4-dimethoxybenzylamine) -3- was washed with saturated aqueous NaCl (500 mL) and separated. The organic phase was dried over Na 2 SO 4, filtered and concentrated in vacuo to afford 4.33 g of the title compound.

Aminobenzonitrile (3.9 g) in acetonitrile (100 mL) was cooled to 0 ° C and treated with potassium carbonate (6.3 g), followed by a solution containing 3 g of 2-chloro-5-nitro-4-thiocyanatopyrimidine (WO 2003/032994) in acetonitrile (50 mL). The mixture was stirred for 30 minutes at 0 ° C and 30 minutes at room temperature, resulting in the formation of a precipitate. The mixture was cooled to 0 ° C by the addition of 4% acetic acid (150 mL) and filtered. The precipitate was stirred by rotation in 100 mL of acetonitrile and filtered again. The precipitate was washed with acetonitrile, which resulted in slow dissolution of the product in the filtrate. After air drying 1.5 g of the title compound remained as the precipitated cake. The filtrate was extracted with EtOAc, dried over Na 2 SO 4, filtered and concentrated in vacuo. Column chromatography (0 → 20% EtOAc / DCM) and recrystallization from acetonitrile provided an additional 0.415 g of the title compound.

OMe

NO2

4- (2,4-dimethoxybenzylamine) -3- (5-nitro-4-thiocyanatopyrimidin-2-ylamine) benzonitrile

A solution of 4- (2,4-dimethoxybenzylamine) -3- (R) -4- (2,4-dimethoxybenzylamine) -3- (4- (6,8-difluorochroman-4-ylamine) -5-nitropyrimidine-1 2-ylamino) benzonitrile

A partial suspension of 4- (2,4-

dimethoxybenzylamine) -3- (5-nitro-4-thiocyanatopyrimidine-2-one

(415 mg) in 40 mL of acetonitrile was treated with a solution of HCl (R) -6,8-difluorochroman-4-amine salt (320 mg) in DMSO (10 mL), followed by potassium carbonate. (1.0 g). The mixture was stirred for 24 hours and then diluted with 10 EtOAc (200 mL). The mixture was washed once with saturated aqueous ammonium chloride (200 mL) and 3 times with saturated aqueous NaCl (200 mL each). The organic layer was separated, dried over Na 2 SO 4, filtered and concentrated in vacuo. Column chromatography (20-40% EtOAc / hexanes) provided 358 mg of the title compound.

(R) -4- (2,4-dimethoxybenzylamine) -3- (9- (6,8-

difluorochroman-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-ylamine) benzonitrile

A solution of (R) -4- (2,4-dimethoxybenzylamine) -3- (4- (6,8-difluorochroman-4-ylamine) -5-nitropyrimidin-2-ylamine) benzonitrile (358 mg) in THF ( 25 mL) was treated with a solution of sodium hydrosulphite (1.5 g) and sodium bicarbonate (1.5 g) in 20 mL of distilled water. Methanol (5 mL) was added to keep the solution homogeneous. After 15 minutes, the mixture was diluted with EtOAc (100 mL) and washed with saturated aqueous NaCl (2 x 100 mL). The organic layer was separated, dried over Na 2 SO 4, filtered and concentrated in vacuo to afford intermediate (R) -4- (2,4-dimethoxybenzylamine) -3- (5-amino-4- (6,8-difluorochroman-4 - ylamine) pyrimidin-2-ylamine) benzonitrile. The intermediate was dissolved in THF (5 mL) and treated with carbonyldiimidazole (0.55 g) for 16 hours. The mixture was diluted with EtOAc (100 mL) and washed twice with saturated aqueous NaCl (2 x 100 mL). The organic layer was separated, dried over Na 2 SO 4, filtered and concentrated in vacuo. Column chromatography (2-3% MeOH / DCM) provided 230 mg of the title compound.

(9- (6,8-Difluorochroman-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-ylamine) benzonitrile (230 mg) in DCM (5 mL) was treated with TFA (5 mL) and triethyl silane (1 mL) for 16 h. The mixture was concentrated to

difluorochroman-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-ylamine) benzonitrile. The intermediate was dissolved in 5 mL of THF and treated with 3 mL of trimethylortoformate, followed by 15 p-toluenesulfonic acid (3 mg). After 1 hour, the mixture was diluted with EtOAc (100 mL) and washed once with saturated aqueous sodium bicarbonate (100 mL). The organic layer was separated, dried over Na 2 SO 4, filtered and concentrated in vacuo. Column chromatography (50 → 100% EtOAc / hexanes) provided 78 mg of the title compound. 1H-NMR (300 MHz, 5% CD3OD in CDCl3) δ 8.8 (s, 1H), 8.7 (s,

F

5th

3- (9 - ((R) -6,8-difluorochromano-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-3 5-carbonitrile

A solution of (R) -4- (2,4-dimethoxybenzylamine) -3-

vacuum to provide intermediate (R) -4-amino-3- (9- (6,8-

1H), 8.2 (s, 1H), 7.8 (d, 1H), 7.6 (dd, 1H), 6.8 (td, 1H), 6.4 (dd, 1H), 5, Δ (dd, 1H), 4.6 (m, 1H), 4.4 (td, 1H), 2.9 (m, 1H),

2.3 (m, 1H). 3- (9- ((R) -chromano-4-yl) -8-οχο-8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-5-carbonitrile

The title compound was synthesized from (R) -chroman-4-amine by the procedures described in Example 26.

3- [9- (8-fluoro-chroman-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-yl] -3H-benzoimidazole-5-carbonitrile

The title compound was synthesized from (R) -8-fluorochroman-4-amine by the procedures described in

3- (9- ((R) -6-fluorochroman-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-5-one carbonitrile

The title compound was synthesized from (R) -6-fluorochroman-4-amine by the procedures described in Example 26. 1H NMR (300 MHz, CDCl3 + 5% CD3OD): δ 8.86 (s, 1H ),

Conditions for introduction of chromanil amine were improved as described below:

1H-NMR (300 MHz, 5% CD3OD in CDCl3) δ 8.8 (s, 1H), 8.5 (s, 1H), 8.2

(s, 1H), 7.8 (4 1H), 7.5 (dd, 1H), 7.1 (m, 2H), 6.8 (d, 1H), 6.7

(td, 1H), 5.8 (dd, 1H), 4.5 (m, 1H), 4.3 (td, 1H), 2.8 (m, 1H),

2.3 (m, 1H).

F

NC

H

Example 26. 1H-NMR (300 MHz, CDCl3) δ 8.8 (s, 1H), 8.6 (s, 1H), 8.2 (s, 1H), 7.8 (d, 1H), 7 6.6 (d, 1H), 7.0 (t, 1H), 6.6 (m, 2H), 5.8 (t, 1H), 4.6 (m, 1H), 4.4 (m, 1H), 2.8 (m, 1H), 2.4 (m, 1H).

8.41 (s, 1H), 8.18 (s, 1H), 7.72 (d, 1H), 7.51 (d, 1H), 7.0-7.1 (m, 1H), 6 , 8.6-6.9 (m, 1H), 6.49 (dd, 1H), 5.76 (br t, 1H), 4.4-4.5 (m, 1H), 4.24 (br t , 1H), 2.7-2.9 (m, 1H), 2.2-2.3 (m, 1H). Thee laugh5) Thee f ° v

IfV ^ NH η ρ | fV ~ NH ο V *

Meoj ^ A-nYVcn nh-2 ^ - X »ynJih

L '^ νΝ02 DlEA DMSO n ^ vNO9 CN CN

(R) -4- (2,4-dimethoxybenzylamine) -3- (4- (6-fluorochroman-4-ylamine) -5-nitropyrimidin-2-ylamine) benzonitrile

A solution of 4- (2,4-dimethoxybenzylamine) -3- (5-nitro-4-thiocyanatopyrimidin-2-ylamine) benzonitrile (139 mg) in anhydrous DMSO (3 mL) was added to a solution of (R ) -6-fluorochroman-4-amine (79 mg) in anhydrous DMSO (3 mL) and DIEA (0.21 mL), the resulting dark red solution being stirred at RT under an air atmosphere and during that time the solution brightened to yellow. At the conclusion of the reaction, the mixture was cooled to 0 ° C with an ice bath and water (25 mL) was added (exotherm). The resulting yellow solid was collected by filtration, washed with additional water, air dried, then dissolved in CH 2 Cl 2, the organic solution being dried (MgSO 4), filtered and evaporated to obtain the title compound (quant.). NMR CDCl 3 1H δ 9.0 (s, 1H), 8.6 (d, 1H), 7.7 (br s, 1H), 7.4 (dd, 1H),

7.1 (d, 1H), 7.0-6.8 (m, 4H), 6.5-6.4 (m, 2H), 5.2 (br s, 1H), 4.3 (s , 2H), 4.2 (br s, 2H), 3.8 (s, 6H), 2.2 (br s, 1H), 1.8 (br s, 1H); 19F δ -123 ppm; MH + = 572.

This material was taken using the same procedures outlined in Example 26 to give 3- (9 - ((R) -6-fluorochroman-4-yl) -8-oxo-8,9-dihydro-7H-purine 2-yl) -3H-benzo [d] imidazole-5-carbonitrile.

3- (9 - ((R) -7-fluorochroman-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-5-one carbonitrile

The title compound was synthesized from

(R) -7-fluorochroman-4-amine by the procedures described in 10

15

20

Example 26. 1 H NMR (300 MHz, CDCl 3 + 5% CD 3 OD): δ 8.86 (s, 1H), 8.58 (s, 1H), 8.19 (s, 1H), 7.79 (d , IH), 7.56 (d, 1H), 6.7-6.9 (m, 2H), 6.4-6.5 (m, 1H), 5.78 (br t, 1H), 4 , 5-4.6 (m, 1H), 4.32 (br t, 1H), 2.7-2.9 (m, 1H), 2.2-2.4 (m, 1H).

NO H

3- [9- (5,8-Difluoro-chroman-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-yl] -3H-benzoimidazole-5-carbonitrile

The title compound was synthesized from (R) -5,8-difluorochroman-4-amine by the procedures described in Example 26. H-NMR (300 MHz, CDCl 3) δ 8.8 (s, 1H), 8 , 7 (s, 1H), 8.2 (s, 1H), 7.8 (d, 1H), 7.6 (d, 1H), 7.0 (m, 1H), 6.4 (m, 1H), 5.9 (t, 1H), 4.6 (m, 1H), 4.4 (m, 1H), 2.5 (m, 2H).

Example 27: Synthesis of 2- (6-Fluoro-1H-benzo [d] imnidazola-1-yl) -9 - ((R) -8-fluoro-chroman-4-yl) -7H-purine-8 (9H ) -ona

Jp = Y NH 2 (BOC) 2 O

So2 dmap, dcm f

TFA -i

DCM

BOC

NH

Na2S2O4ZNaHCO3

NO2 THF / H2O / MeO H F

.Cc

BOC

NH

NH2

4-Fluoro-2-nitro-phenyl-di-tertiary-butyl imidodicarbonate

A catalytic amount of DMAP was added to a mixture of 4-fluoro-2-nitrobenzenamine (0.78 g) and di-tertiary butyl dicarbonate (2.18 g) in DCM (20 mL) and stirred at room temperature for 3 pm The mixture was diluted with H 2 O and twice extracted with DCM, the combined organics were dried, filtered and evaporated to obtain bis-BOC material (quant.). 1H-NMR (300 MHz, CDCl3) δ 7.8 (dd, 1H), 7.3 (m, 2H), 1.4 (s, 18H).

Tertiary Butyl 4-Fluoro-nitrophenylcarbamate

(Procedure: Connell, R.D .; Rein, T .; Akermark,

B.; Helquist, P.J., J. Org. Chem., 1988, 55, 3845)

To a stirred solution of Bis-BOC material in DCM (20 mL) was added TFA (0.58 mL). After 3 h the reaction was cooled with aqueous NaHCO 3 (5 mL), brine was added and the mixture was separated and extracted with additional DCM. The combined organics were evaporated, purified by column chromatography (elution with 7.5% EtOAc / Hexane) to give the title product (1.12 g). 1H-NMR (300 MHz, CDCl3) δ 9.5 (br 1H), 8.5 (dd, 1H), 7.9 (dd, 1H), 7.3 (m, 1H), 1.5 (s 9H).

Tertiary Butyl 2-amino-4-fluorophenylcarbamate

To a solution of tert-butyl 4-fluoro-2-nitrophenylcarbamate (0.34 g) in THF (30 mL) was added a premixed solution of sodium hydrosulfite (2 g) and sodium bicarbonate (1 g). in water (50 mL). MeOH (10 mL) was also added to aid the solution of the mixture, which was stirred at room temperature for 30 min, when sodium chloride was added to saturate the solution. The resulting mixture was extracted with EtOAc (2x). The combined organics were dried, filtered and evaporated to obtain the title compound (quant.), Which was used as such for the next step. 1H-NMR (300 MHz, CDCl3) δ 7.5 (dd, 1H), 6.6 (dd, 1H), 6.5 (m, 1H), 6.4 (br, 1H), 4.7 ( br, 2H), 1.5 (s, 9H); MH + = 227 (minor) 127 (-BoC), 171 (-tBu).

BOC

Λη

Ji

"NH2

! N Cl γγγ; ΡΙ ___,

N ^ sNO, ΕΐσΗ Ν '^ -' ΝΟο K2CO3i ACN

BOC

2-Chloro-5-nitro-4-thiocyanatopyrimidine

(Compound already known, for example, in patent

WO 2003/032994) Potassium thiocyanate (0.97 g, 10 mM) was added to a solution of cooled to 2,4-dichloro-5-nitropyrimidine (1.94 g, 10 mM) in EtOH (40 mL). 0 ° C by an ice bath. The solution was stirred at 0 ° C for 30 min, then the bath was removed and the resulting suspension allowed to reach RT for 60 min when water (100 mL) was added. The precipitate was collected by filtration, washed with ice cold water, dissolved with DCM, dried (MgSO 4), filtered and evaporated to obtain the title compound (1.7 g). 1H-NMR (300 MHz, CDCl3) δ δ 9.4 (s, 1H).

Tertiary Butyl 4-fluoro-2- (5-nitro-4-

thiocyanatopyrimidin-2-ylamine) phenyl carbamate

Potassium carbonate (207 mg) was added to a stirred solution of 2-chloro-5-nitro-4-thiocyanatopyrimidine (108 15 mg) and tertiary butyl 4-fluoro-2-nitrophenyl carbamate (113 mg) in ACN (5 mL). ) and stirred for 15 h. The solution was diluted with brine and extracted with EtOAc (2x). The combined organics were evaporated and purified by column chromatography; elution with 30% EtOAc / Hex gave the title compound (144 mg, 71% yield). 1H-20 NMR (300 MHz, DMS0-d6) δ 10.5 (br s, 1H), 9.3 (br s, 1H), 8.9 (br s, 1H), 7.7-7.4 (m, 2H), 7.1 (br s, 1H), 1.5 (s, 9H), 1.5 (s, 9H); MH + = 407, 307 (-BoC), 351 (-tBu).

F

30CW

1 F / NaHCOg BQC_ ^ Φ

THF / H2O / MbOH

V iAo1 Vr

/ NaHCOg BOC_ ^ CDt

J N0 Xn0j KaCO3. ACN

NH

H2

THF

rO> _f j q F

VO {"Ή N VO ^ tiioh Ç

DO ”orjcv ** ■ *>

M CH FH

V

F

(R) -Tertiary Butyl 4-fluoro-2- (4- (8-fluorochrome

4-ylamine) -5-nitropyrimidin-2-ylamine) phenylcarbamate

A solution of (R) -8-fluorochroman-4-amine

hydrochloride (104 mg) in DMSO (2 mL) and potassium carbonate (141 mg) were added to a stirred solution of tertiary butyl 4- fluoro-2- (5-nitro-4-thiocyanatopyrimidin-2-ylamine) phenyl 1 carbamate (140 mg) in ACN (10 mL). The mixture was stirred for 15 h at room temperature, then partitioned between brine and EtOAc and separated. The heated layer was washed with additional 5 EtOAc, the combined organics were evaporated and purified by column chromatography; elution with 20-30% EtOAc / H gave the title product in 83% yield. 1H-NMR (300 MHz, CDCl3) δ

9.1 (s, 1H), 8.7 (m, 1H), 8.2 (br s, 1H), 7.7 (m, 1H), 7.3 (m, 1H),

7.3-6.8 (m, 4H), 6.5 (s, 1H), 5.5 (br s, 1H), 4.4 (m, 2H), 2.4 (m, 1H), 2.2 (m, 1H), 1.5 (s, 9H); MH + = 515.459 (-tBu).

(R) -Tertiary Butyl 4-fluoro-2- (9- (8-fluoroeromano

4-yl) -8-OXO-8,9-dihydro-7H-purine-2-ylamine) phenylcarbamate

To a solution of (R) -tertobutyl 4-fluoro-2- (4- (8-fluorochroman-4-ylamine) -5-nitropyrimidin-2-15-ylamine) phenylcarbamate (141 mg) in THF (20 mL) A premix solution of sodium hydrosulfite (0.6 g) and sodium bicarbonate (0.3 g) in water (50 mL) was added. MeOH (5 mL) was also added to aid the solution of the mixture, which was stirred at room temperature for 30 min, when sodium chloride was added to saturate the solution. The resulting mixture was extracted with EtOAc (2x) and the combined organics were dried, filtered and evaporated to give 2- (5-amino-4- (8-fluorochroman-4-ylamine) pyrimidine-2-R (tert-butyl) pyrimidine-2 -ylamine) -A-

fluorophenyl carbamate, which was used as such for the next step, MH + = 485.

To a stirred solution of the above material in THF (5 mL) was added CDI (131 mg). After 15h, brine and EtOAc were added and the mixture was separated. The heated layer was washed with additional EtOAc and the combined organics were evaporated and purified by column chromatography (elution with 3% MeOH / DCM) to obtain the title product (86 mg, 62% yield for two steps). . 1H-NMR (300 MHz, 5% CD3OD in CDCl3) δ 7.9 (s, 1H), 7.4 (dd, 1H), 7.3 (m, 1H), 6.9 (dd, 1H) ,

6.7-6.5 (m, 3H), 5.7 (dd, 1H), 4.6 (m, 1H), 4.3 (td, 1H), 2.9 (m, 1H), 2 1.2 (m, 1H); 1.5 (s, 9H); MH + = 511, 411 (-BoC), 455 (-tBu).

2- (6-fluoro-1H-benzo [d] imidazola-1-yl) -9 - ((R) -8-fluoroeromano-4-yl) -7H-purine-8 (9H) -one Preparation of 30% TFA / DCM (5 mL) was added to (R) -Tertiary butyl 4-fluoro-2- (9- (8-fluorochroman-4-yl) -8-oxo-8,9-di -hydro-7H-purin-2-ylamine) phenylcarbamate and the solution was stirred at room temperature for 560 min, then solvents were removed in vacuo to obtain (R) -2- (2-amino-5-fluorophenylamino) ) -9- (8-fluorochroman-4-yl) -7H-purine-8 (9H) -one, which was used as such; MH + = 411.

To the above diamine were added MeOH (2 mL), trimethylortoformate (2 mL) and p-TsOH (cat.). The mixture was stirred at RT at 60 ° C, then the solvents were reduced and the resulting material partitioned between DCM and brine and separated. The crude product was purified by column chromatography (elution with 4% MeOH / DCM) to obtain the title compound (46 mg). 1H-NMR (300 MHz, 5% CD3OD in CDCl3) δ 8.7 (s, 1H), 8.1 (s, 1H), 7.5 (m, 15 2H), 6.9 (m, 2H ), 6.6 (m, 2H), 5.8 (dd, 1H), 4.6 (m, 1H), 4.3 (td, 1H), 2.8 (m, 1H), 2.3 (m, 1H); MH + = 421.

2- (6-Fluoro-1H-benzo [d] imidazola-1-yl) -9 - ((R) -6-fluorochromano-4-yl) -7H-purine-8 (9H) -one

The title compound was synthesized from (R) -6-fluorochroman-4-amine by the procedures described in Example 27. 1 H NMR (300 MHz, CDCl 3 + 5% CD 3 OD): δ 8.72 (s, 1H ), 8.16 (s, 1H), 7.5-7.7 (m, 2H), 6.9-7.0 (m, 2H), 6.8-6.9 (m, 1H), 6.54 (dd, 1H), 5.78 (br t, 1H), 4-4.5 (m, 1H), 4.26 (m, 1H),

2.7-2.8 (m, 1H), 2.2-2.3 (m, 1H). 2- (6-fluoro-1H-benzo [d] imidazola-1-yl) -9- (tetrahydro-2H-pyran-4-yl) -7H-purine-8 (9H) -one

The title compound was synthesized from 4-amino-tetrahydropyran by the procedures described in Example 5. 27. 1H NMR (d6-DMS0) δ 11.65 (s, 1H), 9.13 (s, 1H) , 8.34 (s, 1H), 8.28 (m, 1H), 7.82 (m, 1H), 7.25 (td, J = 9.0, 2.4 Hz, 1H), 4, 56 (m, 1H), 4.03 (dd, J = 11.1, 3.9 Hz, 2H), 3.50 (t, J = 11.1 Hz, 2H), 2.59 (m, 2H ), 1.78 (m, 2H).

2- (6-Chloro-1H-benzo [d] imidazola-1-yl) -9- ((R) -8-fluorochromano-4-yl) -7H-purine-8 (9H) -one

The title compound was synthesized from (R) -8-fluorochroman-4-amine and 4-chloro-2-nitrobenzenamine by the procedures described in Example 27. 1H NMR (300 MHz, CDCl3 + 5% CD3OD): δ 8.7 (s, 1H), 8.2 (s, 1H), 8.1 (s, 1H), 7.6 (d, 1H), 15 7.2 (dd, 1H), 6.9 (td, 1H), 6.7-6.5 (m, 2H), 5.8 (dd, 1H), 4.6 (m, 1H), 4.4 (td, 1H), 2.9 ( m, 1H), 2.3 (m, 1H).

Example 28: Synthesis of 2- (5-Fluoro-1H-beinzo [d] imidazola-1-yl) -9- ((R) -8-fluorochroman-4-yl) -7H-purine-8 (9H) - one

20 N- (2,4-Dimethoxybenzyl) -5-fluoro-2-

nitrobenzenamine

A solution of 2,4-difluoro-1-nitrobenzene (1.1 mL), 2,4-dimethoxy benzylamine (1.5 mL) and DIEA (5.2 mL) in THF (40 mL) was heated to 60 ° C for 60 min; allowed to cool at RT, partitioned between EtOAc and H 2 O, separated, dried (MgSO 4), filtered and evaporated to obtain the title product as a yellow solid (3.14 g). 1H-NMR (300 MHz, CDCl3) δ 8.5 (br s, 1H), 7.2 (dd, 1H), 7.2 (d, 1H), 6-6-6.4 (m, 3H) , 6.3 (m, 1H), 4.3 (d, 2H),

3.9 (s, 3H), 3.8 (s, 3H).

N1- (2,4-Dimethoxybenzyl) -5-fluorobenzene-1,2-

diamine

Under air heating, a catalytic amount

of a solution of Raney NI in water was added to a solution of N- (2,4-dimethoxybenzyl) -5-fluoro-2-nitrobenzenamine (0.5 g) in THF (20 mL). The flask was closed with a septum, evacuated under vacuum and hydrogen was added by means of a flask. The resulting suspension was stirred at RT for 16 h, when the H2 flask was removed and the mixture was evacuated and filtered through a celite plug, which was rinsed thoroughly with THF and MeOH to obtain the titled diamine, which was used as such. .

(R) -2-Chloro-N- (8-fluorochroman-4-yl) -5-nitropyrimidine-4-amine

A solution of (R) -8-fluorochroman-4-amine hydrochloride (1.02 g) and DIEA (2.6 mL) in DCM (10 mL) was slowly added to a solution of 2,4-dichloro-5- nitropyrimidine (0.97 g) and THF (25 mL) at -78 ° C. The reaction mixture was stirred for 30 min at -78 ° C, then allowed to warm overnight at RT. The reaction was cooled with the addition of saturated NH 4 Cl (1 mL), the solvent volume was reduced in vacuo and the resulting mixture partition between EtOAc and water was then separated. The crude material was purified by column chromatography and elution with 30% EtOAc / Hex gave the title product (1.43 g). 1H-NMR (300 MHz, CDCl3) δ 9.1 (s, 1H), 8.6 (br d, 1H), 7.1-6.8 (m, 3H), 5.6 (dd, 1H) , 4.4 (m, 1H), 4.3 (m, 1H), 2.4 (m, 1H), 2.2 (m, 1H).

(R) -N2- (2- (2,4-dimethoxybenzylamine) -4-fluorophenyl) -N4- (8-fluorochromano-4-yl) -5-nitropyrimidine-2,4-diamine

A mixture of (R) -2-chloro-N- (8-fluorochroman-4-yl) -5-nitropyrimidine-4-amine (32 mg), N1- (2,4-dimethoxybenzyl) -

5-Fluorobenzene-1,2-diamine (28 mg) and KCO 3 (41 mg) in ACN was heated at 65 ° C for 3 h, cooled to RT, diluted with brine and extracted with EtOAc (2x). The combined organics were evaporated and purified by column chromatography (elution with 30% EtOAc / Hex) to obtain the title product (21 mg). 1H-NMR (300 MHz, CDCl3) δ 9.0 (s, 1H), 8.6 (br d, 1H), 7.2-6.8 (m, 6H), 6.5-6.3 ( m, 4H), 4.4-4.2 (m, 4H), 3.8 (s, 6H), 2.3-2.2 (m, 2H).

(R) -2- (2- (2,4-Dimethoxybenzylamine) -4-fluorophenylamine) -9- (8-fluorochromano-4-yl) -7H-purine-8 (9H) -one

Under an air atmosphere, a catalytic amount of a Raney NI solution in water was added to a solution of (R) -N2- (2- (2,4-dimethoxybenzylamine) -4-fluorophenyl) -N4- (8-fluorochroman-4-yl) -5-nitropyrimidin-2,4-diamine (21 mg) in THF. The flask was closed with a septum, evacuated under vacuum and hydrogen was added by means of a flask. The resulting suspension was stirred at RT for 2 h and when the H2 flask was removed, the mixture was evacuated and filtered through a celite plug, which was thoroughly rinsed with THF and MeOH to obtain (R) -N2- (2- (2,4-dimethoxybenzylamine) -4-fluorophenyl) -N4- (8-fluorochroman-4-yl) pyrimidin-2,4,5-triamine, which was used directly.

To a stirred solution of the above material in THF (5 mL) was added CDI (12 mg). After 18 h, brine and EtOAc were added and the mixture was separated. The organic layer was evaporated and purified by column chromatography (elution with 4% MeOH / DCM) to obtain the title product (14 mg). 1H NMR (300 MHz, CDCl3 + 5% CD3OD): δ 7.8 (s, 1H), 7.3 (s, 1H), 7.1 (d, 1Η), 6.9 (m, 2Η) , 6.7-6.2 (m, 6Η), 5.7 (dd, 1Η), 4.5 (m, 1Η), 4.2 (m, 1Η), 4.1 (s, 2Η), 3.8 (s, 3Η), 3.7 (s, 3Η), 2.8 (m, 1Η), 2.2 (m, 1Η).

2- (5-Fluoro-1H-benzo [d] imidazola-1-yl) -9 - ((R) -8-

fluorochroman-4-yl) -7H-purine-8 (9H) -one

An

mixture

in

(R) -2- (2- (2.4-

dimethoxybenzylamine) -4-fluorophenylamino) -9- (8-fluorochroman-4-

yl) -7H-purine-8 (9H) -one (14 mg) and TFA (1 mL) was stirred for 60 min when triethyl silane (0.5 mL) was added. The resulting solution was stirred at RT for 16h, then the solvents were reduced under vacuum to obtain (R) -2- (2-amino-4-fluorophenylamino) -9- (8-fluorochroman-4-yl) - 7H-purine-8 (9H) -one, which was used as

added to a solution of the above amine in trimethylortoformate (2 mL). The mixture was stirred at RT for 15h, then the solvents were reduced and the resulting material partitioned between DCM and brine and separated. The crude product was purified by column chromatography (elution with 5% MeOH / DCM) to obtain the title compound (9 mg). 1H-NMR (300 MHz, CDCl3) δ 10.0 (s, 1H),

such.

A catalytic amount of p-TsOH was

8.9 (s, 1H), 8.3 (s, 1H), 7.8 (dd, 1H), 7.4 (d, 1H), 7.1 (m, 2H),

6.8 (m, 2H), 5.9 (dd, 1H), 4.7 (m, 1H), 4.4 (td, 1H), 2.9 (m, 1H),

2.4 (m, 1H).

Example 29: Synthesis and Resolution of 8-

Fluorochroman-4-amine

F

NaOHZH2O

HO

THE

THE

HO "

3- (2-Fluorophenoxy) propanoic acid A mixture of 2-fluorophenol (15 g), 3-bromopropanoic acid (20 g) and NaOH (11 g) was refluxed in 50 mL of water. The solution was cooled to room temperature and acidified to pH = 2 with 3 M HCl. The resulting precipitate was isolated by filtration to obtain 9.27 g of the title compound as a white solid. The filtrate was extracted 3 times with EtOAc to obtain 2.5 g of less pure compound.

8-Fluorochroman-4-one

Oxalyl chloride (8.79 mL) and 1 drop of DMF 10 were added to a frozen solution of 3- (2-fluorophenoxy) propanoic acid (9.27 g) in DCM (50 mL). The solution was stirred at 0 ° C for 2 hours, then aluminum chloride (7.39 g, 55.42 mM) was added and the solution was stirred for 16 hours at room temperature. The mixture was poured into ice water and extracted three times with DCM. The combined organics were washed with 0.5 M NaOH and brine, then dried, evaporated and purified by column chromatography (elution with 20% EtOAc / Hex) to give the title compound (8.20 g , 98%).

8-fluorochroman-4-amine

A round bottom flask was charged with 8-fluorochroman-4-one (8.2 g), hydroxylamine hydrochloride (3.78 g) and sodium acetate (4.46 g). A reflux condenser was added, the flask was purged with argon, dry EtOH (20 mL) was added and the mixture was stirred at reflux for 18 hours. The solution was cooled to room temperature, diluted with EtOAc and washed with water. The organic phase was dried and evaporated to give intermediate 8-fluorochroman-4-amine oxime, which was reduced with Raney Nickel in 50 psi EtOH to obtain the title amine 30 (4.69 g, 57%). F

F

F

NH2

Novozyme 435

MeOCH2COOMe

t-BuOMe

THE

8 M HCt EtOH

F

NB *

8-fluorochroman-4-amine resolution

(Procedure based on US patent 0157739)

A mixture of 8-fluorochroman-4-amine (3.40 g)

Methyl 2-methoxy acetate (2.44 g) and Novozyme 435 (Aldrich, 0.68 g) in anhydrous methyl tertiary butyl ether (75 mL) was heated at reflux under argon for 2 hours (at this time from acylated to non-acylated product was 1: 1 by HPLC). The solid that formed on cooling was collected by filtration and dissolved in EtOAc. The mixture was filtered to remove the catalyst and washed once with 0.5 M HCl to remove any remaining (S) -amine. The solvent was evaporated and the product was recrystallized from tertiary butyl methyl ether to obtain (R) -N- (8-fluorochroman-4-yl) -2-methoxy-acetamide (0.78 g). The reaction solvent and the recrystallization mother liquor were washed 3 times with 0.5 M HCl and concentrated to give additional (R) -N- (8-fluorochroman-4-yl) -2-methoxy acetamide ( 0.83 g). The combined acidic aqueous layers were made basic by NaOH and extracted with DCM to obtain (S) -8-fluorochroman-4-amine (1.6 g). A solution of (R) -N- (8-fluorochroman-4-yl) -2-methoxyacetamide (0.78 g) in 8 M HCl in EtOH (50 mL) was heated at reflux for 4 hours. The solvents were removed from the cooled reaction mixture, the resulting solid was taken up in 50 mL of 0.5 M NaOH, salted with NaCl and extracted 4 times with DCM to obtain (R) -8-fluorochroman-4-amine ( 0.48 g (87%)). The% ee was determined by chiral HPLC: Chiralcel OD-H method (0.46 x 25 cm analytical column, Daicel Chemical Industries): isocratic, 5% (0.05% TFA / ETOH), 95% (0 0.05% TFA / HEX), R t = 7.2 min, (S) enantiomer, R t = 9.2 min, (R) enantiomer.

methylchroman-4-amine, 6-methoxychroman-4-amine, 7-fluorochroman-4-amine, 5,8-difluorochroman-4-amine and 6,8-difluorochroman-4-amine

described in Example 29 for the synthesis of 8-fluorochroman-4-amine. Corresponding chromano-4-ones were commercially available as advanced intermediates for the synthesis of chromano-4-amine, 6-fluorochromano-amine, 6-chlorochromano-4-amine, 6-methylchromano-4-amine and 6-methoxyromano-4. -the mine. For the synthesis of 5-fluorochroman-4-amine, intermediate 5-fluorochroman-4-one was obtained using GB 2355264 procedures, which also provided 7-fluorochroman-4-one. 7-Fluorochroman-4-one could be used in the synthesis of 7-fluorochroman-4-amine. The chroman-4-amine, 5-fluorochroman-4-amine, 6-fluorochroman-4-amine, 7-fluorochroman-4-amine, 5,8-difluorochroman-4-amine and 6,8-difluorochroman-4-amine were resolved by the procedure described in Example 29 for the resolution of 8-fluorochroman-4-amine.

the mine

Example 30: Chroman-4-amine, 5-fluorochroman-4

6-fluorochroman-4-amine, 6-chlorochroman-4-amine, 6-

F

F

These amines were prepared by procedures

Example 31: 1-Methyl-4,5,6,7-tetrahydro-1H

indole-4-amine

NH 2 The title compound was obtained from 1-methyl-6,7-dihydro-1H-indola-4 (5H) -one ("Heterocycles"

(Heterocycles) (1984), 22, 2313), by the procedure described in Example 29, which was used to obtain 8-fluorochroman-4-amine from 8-fluorochroman-4-one.

Example 32: Synthesis of 5,6-Difluorochroman-4

the mine

Br

JL OH BrCH2CH ^ COOH

: -V

NaOH

0— ^ COOH

IJ (Coa) 2 2) Aia3

NH2OH-HO1 NaOAc EIOH1 reflux

1H2 / Raney-Ni 2. H21 Pd / C

3- (2-Bromo-4,5-difluorophenoxy) propanoic acid

A solution of 1.68 g of NaOH (42 mmol) in 5 mL of water was slowly added to a suspension of 2.29 mL (20 mmol) of 2-bromo-4,5-difluorophenol and 3.07 g. (20 mmol) 3-bromopropionic acid. The mixture was heated at 100 ° C in an oil bath for 5 hours and then allowed to cool to room temperature. Water was added to completely dissolve any solid material and the reaction mixture was made acidic with concentrated HCl. The product was extracted into ether (3 times) and the combined organic layers were dried over Na 2 SO 4 and evaporated to give 3.7 g (66%) of the title compound as a light brown solid. 1H NMR (300 MHz, CDCl3): δ 7.4 (t, 1H), 6.8 (q, 1H), 4.3 (t, 2H), 2.9 (t, 2H).

8-Bromo-5,6-difluoro-2,3-dihydrochroman-4-one

Oxalyl chloride (1.7 mL, 20 mmol) was added to a solution of 2.8 g (10 mmol) of 3- (2-bromo-4,5-difluorophenoxy) propanoic acid in 40 mL of anhydrous DCM, followed by 25 a drop of DMF. After 1.5 hours, a drying tube was coupled and the solution was cooled in a frozen water bath. AlCl 3 (1.5 g, 11 mmol) was added and the dark red solution was allowed to reach room temperature slowly and stirred for 16 hours. The mixture was poured into ice and the organic layer was separated. The aqueous layer was extracted with DCM twice. The combined organic layers were washed with 0.5 N NaOH and brine, then dried over Na 2 SO 4 and concentrated. Column chromatography of this residue with hexane and EtOAc provided 1.9 g of the title compound as an off-white solid (73%). 1H NMR (300 MHz, CDCl3): δ 7.6 (t, 1H), 4.65 (t, 2H), 2.85 (t, 2H).

8-Bromo-5,6-difluoro-2,3-dihydrochroman-4-one

oxime

To a solution of 8-bromo-5,6-difluoro-2,3-dihydrochroman-4-one (7.2 mmol) in 40 mL of ethanol was added hydroxylamine hydrochloride (0.55 g, 7.9 mmol ) and sodium acetate (0.65 g, 7.9 mmol). This mixture was heated at reflux for 20 h.

The mixture was cooled, diluted with EtOAc, washed with water and brine and then dried with Na 2 SO 4. Concentration of the solvent provided the title compound as a white solid (1.9 g). 1H NMR (300 MHz, 10% CD3OD in CDCl3): δ 7.3 (t, 1H), 4.2 (t, 2H),

2.9 (t, 2H).

5,6-Difluoro-3,4-dihydro-2H-chroman-4-amine

Raney-NI (5 mL of sludge in water) was added to a solution of 8-Bromo-5,6-difluoro-2,3-dihydrochroman-4-one oxime (1.9 g) in 200 mL of MeOH . The mixture was hydrogenated at 50 psi for 24 h to provide 8-bromo-5,6-difluoro-3,4-dihydro-2H-

chroman-4-amine. Pd / C (0.3 g) was added to the mixture and hydrogenation was resumed at 50 psi for 4 hours. The title compound was obtained after filtration and concentration in vacuo. 1H NMR (300 MHz, 10% CD3OD in CDCl3): δ 7.15 (q, 1H), 6.6 (m, 1H), 4.6 (bm, 1H), 4.25 (bm, 2H), 2.2-2.4 (m, 2H).

Example 33: Synthesis of 4-Amino-3,4-dihydro-2H-

chroman-8-carbonitrile

CN ÇN NH4OAc / NaCNBH3

WIeOH / sieves 3A

NH2

4-Amino-3,4-dihydro-2H-chroman-8-carbonitrile A mixture of 260 mg of 4-oxo-3,4-dihydro-2H-chroman-8-carbonitrile (made from 2 -hydroxybenzonitrile by the procedure described in Example 29), ammonium acetate (1.2 g) and 3A molecular sieves (1.5 g) in 10 mL methanol was stirred for 5 days. The mixture was filtered through celite and the filtrate concentrated in vacuo. The crude residue was treated with 100 mL 1 M HCl and extracted with ethyl ether (3 x 100 mL). The aqueous layer was made basic to pH = 10 with saturated NaOH and extracted with DCM (3 x 100 mL). The combined DCM layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo to afford 150 mg of the title compound.

NC NH2

4-Amino-3,4-dihydro-2H-chroman-6-carbonitrile

The title compound was made from 6-cyano-

4-chromanone (Syntech) by the same procedure as described in Example 33.

NH2

4-Amino-1,2,3,5,4-tetraacetate

hydronaphthalene-1-yl

The title compound was made from 4-oxo-1,2,3,4-tetrahydronaphthalen-1-yl acetate [Tetrahedron:

Assimetry, 2001, 12, 2283) by the same procedure as described in Example 33.

, N,

H2N

6,7-Dihydro-5H-cyclopenta [b] pyridine-5-amine

The title compound was made as described in WO 03/045924. Example

hydroquinoxaline-5-amine Boc2O

34

DMAP '[N

ch ^ cn Boc "NY" O

Synthesis

H2NNH2

in

(R) -5,6,7,8-Tetra-

MeOH

Mouth

TFA

CH2CI2

90

NH2

acetyl (5,6,7,8-tetramethyl

(R) -Tertiary Butyl Hydroquinoxaline-5-yl) Carbamate

A solution containing 483 mg of (R) -N- (5,6,7,8-

tetrahydroquinoxaline-5-yl) acetamide (J. Org. Chem. (2003), 68, 3546) in acetonitrile (20 mL) was treated with Boc20 (3 g) and DMAP (5 mg). The mixture was heated at 60 ° C for 1.5 h and then concentrated in vacuo. Column chromatography (50% EtOAc / hexanes) provided 293 mg of the title compound.

(R) -Tertiary Butyl 5,6,7,8-tetrahydroquinoxaline-

5-ylcarbamate

A solution of (R) -Tertiary butyl acetyl (5,6,7,8-tetrahydroquinoxaline-5-yl) carbamate (293 mg) in methanol (10 mL) 15 was treated with hydrazine hydrate (0.5 mL) for 1.5 h. The mixture was diluted with EtOAc and washed twice with saturated aqueous sodium chloride. The organic layer was separated, dried over sodium sulfate and concentrated in vacuo to afford 238 mg of the title compound.

(R) -5,6,7,8-Tetrahydroquinoxaline-5-amine

A solution of (R) -Tertiary butyl 5,6,7,8-tetrahydroquinoxaline-5-ylcarbamate (238 mg) in 10 ml of 1: 1 TFA / DCM was stirred for 30 minutes. The mixture was concentrated in vacuo to afford the title compound as a TFA salt.

Example 35: Synthesis of 2- (1H-Benzo [d] imidazole-1-amine)

yl) -9- (4,5,6,7-tetrahydro-1H-indola-4-yl) -7H-purine-8 (9H) -one>

Pl

PhSO2Cl, 1,2-di chloroethane NaOH

I>

N

SO2Ph

NH2

W

SO2Ph

N

, SO2Ph

huh?

4 N NaOH MeOH

Ç&

"y-N

XX> = °

1- (Phenylsulfonyl) -4-oxo-4,5,6,7-tetrahydroindole

To a suspension of NaOH (4.44 g) in 1,2-dichloroethane (250 mL) was added 4-oxo-4,5,6,7-tetrahydroindole (5.0 g). The mixture was then cooled to 0 ° C and stirred for min and then a solution of phenylsulfonyl chloride (5.7 mL) in 1,2-dichloroethane (50 mL) was added by dripping over a period of time. 30 min After 30 min stirring, the reaction mixture was allowed to reach room temperature and was stirred overnight. The reaction was cooled by pouring into distilled water (100 mL). The organic layer was separated and the aqueous layer was extracted with dichloromethane (3 x 50 mL). The combined organic extract was washed with distilled water to neutrality, dried over MgSO 4 and concentrated in vacuo to afford 7.0 g of the title compound.

1- (Phenylsulfonyl) -4,5,6,7-tetrahydro-1H-indola-4-amine

The title compound was obtained from 1- (phenylsulfonyl) -4-oxo-4,5,6,7-tetrahydroindole by

procedure described in Example 29, which was used to obtain 8-fluorochroman-4-amine from 8-fluorochroman-4-one.

2- (1H-Benzo [d] imidazola-1-yl) -9- (1-phenylsulfonyl) -4,5,6,7-tetrahydro-1H-indola-4-yl) -7H-purine -8 (9H) - one

The title compound was obtained from 1-

(phenylsulfonyl) -4,5,6,7-tetrahydro-1H-indola-4-amine by

procedures described in Example 24. 10

15

2- (β-benzo [d] imidazola-1-yl) -9- (4,5,6,7-tetrahydro-1H-indola-4-yl) -7H-purine-8 (9H) -one

To a solution of 2- (β-benzo [d] imidazola-1-yl) -9- (1- (phenylsulfonyl) -4,5,6,7-tetrahydro-1H-indol-4-yl ) -7H-purine-8 (9H) -one (50 mg) in MeOH (1 mL) was added 4 N NaOH (1 mL), the mixture was refluxed overnight and cooled. The volatiles were removed under reduced pressure and the resultant neutralized with 4 N HCl. The white precipitate was filtered, washed with a small amount of water and dried in vacuo to afford 36 mg of the title compound. 1H NMR (d6-DMS0) δ 11.6 (s, 1H), 10.7 (s, 1H), 8.86 (s, 1H), 8.27 (s, 1H), 7.69 (d, J = 7.8 Hz, 1H), 7.58 (d, J = 8.1 Hz, 1H), 7.33 (m, 2H), 6.53 (t, J = 2.4 Hz, 1H) , 5.64 (t, J = 2.4 Hz, 1H), 5.54 (m, 1H), 2.72 (m, 2H), 2.30 (m, 1H), 2.07 (m, 2H), 1.84 (m, 1H).

Examples 36 and 37 (Dihydrobenzofuran): 2- (1H-benzo [d] imidazola-1-yl) -9- (2,3-dihydrobenzofuran-3-yl) -7H-purine-8 (9H) -one and 2- (β-benzo [d] imidazola-1-yl) -9- (4-fluoro-2,3-dihydrobenzofuran-3-yl) -7H-purine-8 (9H) -one

(COCO2

NH2OHmci

Not AC

Γ

ÒÒ

.OH

Hg (AI)

THF

Ql 1) 2.5 eq TMSCHN2 2) HOAc

NH2

2-Fluoro-6-methoxybenzoyl chloride

Oxalyl chloride (0.56 mL, 6.4 mmol) was

added to a solution of 1.0 g (5.9 mmol) of 2-fluorochloric acid

6-methoxybenzoic acid in 5 mL of anhydrous CH 2 Cl 2, then a drop of DMF was added. After one hour, when the slow bubbling ceased, the volatiles were removed under reduced pressure to afford 1.1 g (95%) of acid chloride as a pale yellow liquid. 1H NMR (300 MHz, CDCl3): δ 7.45 (q, 1H), 6.7-6.8 (m, 2H), 3.9 (s, 3H).

4-fluorobenzofuran-3 (2H) -one A solution of (trimethyl

yellow silyl) diazomethane (2.0 M, 3.7 mL) was added to 0.57 g (3.0 mmol) of the above acid chloride while stirring. After 3 hours, the solvent was evaporated. The yellow residue was dissolved in 3 mL of acetic acid (strong gas and heat evolution, a water bath was used to cool the flask for one minute) and stirred for 15 min at room temperature. The solvents were removed under vacuum and the red residue was taken up in 2 mL of CH 2 Cl 2, washed twice with water, then brine and dried over Na 2 SO 4. This crude product was purified by column chromatography (elution with 10% EtOAc in hexanes) to give 0.24 g (53%) of 4-fluorobenzofuran-3 (2H) -one as a white solid. 1H NMR (300 MHz, CDCl3): δ 7.58 (m, 1H), 6.92 (br d, 1H), 6.71 (t, 1H), 4.65 (s, 2H).

(Z) -4-fluorobenzofuran-3 (2H) -one oxime

The above ketone (0.70 g, 4.6 mmol) was dissolved in 5 mL of ethyl alcohol and then 0.64 g (9.2 mmol) of hydroxylamine hydrochloride and 0.75 g (9.2 mmol) were added. ) of sodium acetate. This suspension was refluxed for 120 h. The mixture was cooled to room temperature and 4 mL of water was added to dissolve excess reagents. Suction filtration and then washing the solid cake with a small amount of cold water provided 0.48 g (63%) of the desired oxime as white needle-shaped crystals. 1 H NMR (300 25 MHz, CDCl 3): δ 8.38 (s, 1H), 7.38 (q, 1H), 6.6-6.8 (m, 2H), 5.21 (s, 2H) .

4-fluoro-2,3-dihydrobenzofuran-3-amine

The above oxime (0.48 g) was dissolved in 40 mL of anhydrous THF under argon. Freshly prepared aluminum amalgam 30 (by sequentially immersing 1 g of a polished aluminum plate in a 2% aqueous HgCl2 solution, in water and finally in THF) was added rapidly and the mixture was refluxed for 24 hours under Argon. . Shining pellets of mercury appeared at the bottom of the bottle. The mixture was allowed to cool to room temperature and filtered through a pad of celite. The flask and solid cake were washed three times with THF, then three times with methanol. The combined filtrate was evaporated for 10

15

20

rotation to give 0.41 g of a yellow solid as a mixture (determined by NMR) - of approximately 20% of the desired amine with 80% of the initial oxime. This mixture was used for the next step without purification. 1H NMR (300 MHz, CDCl3): δ 7.18 (q, 1H), 6.5-6.7 (m, 2H), 4.8-4.9 (m, 1H), 4.69 (t , 1H), 4.2-4.3 (m, 1H).

2- (β-benzo [d] imidazola-1-yl) -9- (2,3-di-

above racemic hydrobenzofuran-3-yl) -7H-purine-8 (9H) -one may be separated on a chiral CHIRALCEL OD-H column (tris (3,5-dimethylphenylcarbamate cellulose) on a 5 μΜ silica gel substrate eluting with 85:15 hexanes: ethanol (both with 0.1% diethylamine) One enantiomer has a retention time (Tr) of 25 min, the other 33.5 min.

NMR procedures and data for 7-alkylated purinone analogs.

Ioctomethane

PS-BEMP

25

Example 38: 2- (6-Fluoro-β-benzo [d] imidazola-1-yl) -9- ((R) -8-fluorochroman-4-yl) -7-methyl-7H-purine-8 (9H ) -ona

A suspension of 2- (6-fluoro-1H-

benzo [d] imidazola-1-yl) -9 - ((R) -8-fluorochroman-4-yl) -7H-purine-8 (9H) -one (5 mg), 2-tertiary-butylimino-2-one diethylamine-1,3-dimethylperhydro-1,3,3-diazaphosphorine in polystyrene (30 mg, 2.2 mM / g) and iodomethane (5 μΐ in ACN (1 mL) was stirred for 1 h at RT, then filtered through a short plug of celite which was completely rinsed with MeOH Evaporation of solvents and silica gel chromatography (4% MeOH / DCM elution) gave the title compound (4 mg). NMR (300 MHz, CDCl 3) δ 8.7 (s, 1H), 8.2 (s, 1Η), 7.7 (m, 2Η), 7.0 (m, 2Η), 6.7 (m, 2Η), 5.9 (dd, 1Η)

4.7 (m, 1Η), 4.4 (td, 1Η), 3.6 (s, 3Η), 2.9 (m, 1Η), 2.4 (m, 1Η).

NC

H

Br

PS-BEMP

H

/

,THE

Example 39: 3- [9-Chroman-4-yl-7- (2-methoxy)

ethyl) -8-οχο-β, 9-dihydro-7H-purine-2-yl] -3H-benzoimidazole-5-carbonitrile

8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazola-5-carbonitrile (10 mg) in acetonitrile (10 mL) was treated with 2-bromoethylmethyl ether (23 μL) and 2-tertiary-butylimino-2-diethylamine-1,3-dimethyl-perhydro-1,3,2-diazaphosphorin in polystyrene (500 mg,

2.2 mmol base / g). The mixture was stirred at room temperature for 16 hours and then filtered through celite. The celite was washed with acetonitrile and methanol and then the collected filtrate was dried with magnesium sulfate and concentrated in vacuo. Purification by column chromatography (1% MeOH / DCM) gave 4 mg of the title compound. 1H-NMR (300 MHz, CDCl3) δ 8.8 (s, 1H),

yl) -9- ((R) -6-fluorochroman-4-yl) -7- (2-hydroxyethyl) -7H-purine-8 (9H) -one

A solution of 3- (9 - ((R) -chroman-4-yl) -8-oxo

8.6 (s, 1H), 8.4 (s, 1H), 7.9 (d, 1H), 7.6 (d, 1H), 7.2 (m, 2H)

6.9 (d, 1H), 6.8 (d, 1H), 5.9 (t, 1H), 4.6 (m, 1H), 4.4 (m, 1H)

4.2 (m, 2H), 3.8 (m, 2H), 3.4 (s, 3H), 2.8 (m, 1H), 2.4 (m, 1H).

OH

20

Example 40: 2- (6-Fluoro-1H-benzo [d] imidazole-1-10

15

20

25

To a 5 mg solution of 2- (6-fluoro-1H-benzo [d] imidazola-1-yl) -9 - ((R) -6-fluorochroman-4-yl) -7H-purine-8 (9H ) -one in 1 mL of anhydrous acetonitrile was added (excess) 50 mg of 2-tert-butylimino-2-diethylamine-1,3-dimethylperhydro-1,3,2-diazaphosphorine in polystyrene (2.2 mmol base / g, Fluka), then 10 μL (in excess) of 2-iodoethanol. The mixture was stirred at room temperature for 24 hours. Column chromatography with 3% MeOH in CH 2 Cl 2 and then preparative HPLC provided 3 mg of the desired product as a TFA salt. 1H NMR (300 MHz, CDCl3): δ 9.14 (s, 1H), 8.40 (s, 1H), 7.7-7.9 (m, 2H), 7.1-7.2 (m 1H), 7.0-7.1 (m, 1H), 6.9-7.0 (m, 1H), 6.59 (dd, 1H), 5.88 (br t, 1H), 4 , 5-4.6 (m, 1H), 4.32 (t, 1H), 4.0-4.2 (m, 4H), 2.7-2.9 (bs + m, 2H), 2 , 2-2.3 (m, 1H).

HCt

H

PS-BEMP

N-

/

Example 41: 3- [9-Chromano-4-yl-7- (2-dimethylaminoethyl) -8-OXO-8,9-dihydro-7H-purin-2-yl] -3H-benzoimidazole Carbonitrile

A solution of 3- (9 - ((R) -chroman-4-yl) -8-oxo

8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazola-5-carbonitrile (65 mg) in acetonitrile (10 mL) was treated with 2-chloro-N, N-dimethylethylamine hydrochloride (25 mg) and 2-tert-butylimino-2-diethylamine-1,3-dimethyl-perhydro-1,2,2-diazaphosphorine in

polystyrene (325 mg, 2.2 mmol base / g). The mixture was stirred at room temperature for 16 hours and then filtered through celite. The celite was washed with acetonitrile and methanol and then the collected filtrate was dried with magnesium sulfate and concentrated in vacuo. Purification by column chromatography (2% MeOH / DCM) gave 21 mg of the title compound. 1H-NMR (300 MHz, CDCl3) δ 8.9 (s, 1H), 8.5 (s, 1H), 8.4 (s, 1H), 7.8 (d, 1H), 10

15

20

7.60 (d, 1H), 7.2 (m, 2H), 6.9 (d, 1H), 6.8 (d, 1H), 5.9 (t, 1H),

4.4 (m, 4H), 3.6 (m, 3H), 3.0 (s, 6H), 2.2 (m, 1H).

N

NC

TJ>

Br '

"CN

PS-BEMP

, N,

xxy °

NC

N

J

Example 42: 3- (9-Chroman-4-yl-7-cyanomethyl-8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzoimidazole-5-carbonitrile

A solution of 3- (9 - ((R) -chroman-4-yl) -8-oxo

8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-5-carbonitrile

(

(10 mg) in acetonitrile (10 mL) was treated with bromoacetonitrile (17 μυ and 2-tert-butylimino-2-diethylamine-1,3-dimethyl-perhydro-1,3,2-diazaphosphorine in polystyrene (500 mg (2.2 mmol base / g) The mixture was stirred at room temperature for 16 hours and then filtered through celite The celite was washed with acetonitrile and methanol and then the collected filtrate was dried with sulfate Purification by column chromatography (1% MeOH / DCM) gave 11 mg of the title compound 1H-NMR (300 MHz, CDCl3) δ 8.8 (s, 1H), 8.5 (s, 1H), 8.4 (s, 1H), 7.8 (d, 1H), 7.6 (d, 1H), 7.2 (m, 2H), 6.8 (m , 2H), 5.9 (t, 1H), 5.0 (s, 2H), 4.6 (m, 1H), 4.4 (m, 1H), 2.8 (m, 1H), 2 .4 (m, 1H).

Br COOMe

PS-BEMP

MeOOC

Example 43: 2- (2- (6-cyano-1H-benzo [d] imidazola-1-yl) -9- ((R) -6,8-difluorochroman-4-yl) -8-oxo-8, 9-dihydropurin-7-yl) methyl acetate

A solution of 3- (9 - ((R) -6,8-difluorochromano-4-yl) -8-β-8,9-dihydro-7H-purin-2-yl) -3H-benzo [d ] imidazole-5-carbonitrile (10 mg) in acetonitrile (2 mL) was treated with methyl bromoacetate (90 mg) and 2-tertiary-butylimino-2-diethylamine-1,3-dimethyl-perhydro-1,3 Polystyrene 2-diazaphosphorine (100 mg,

2.2 mmol base / g). The mixture was stirred at room temperature.

for 48 hours and then filtered. The collected filtrate was concentrated in vacuo. Purification by preparative TLC (5% MeOH / DCM) followed by column chromatography gave 6 mg of the title compound. 1H-NMR (300 MHz, CDCl3) δ 8.94 (s, 1H), 8.78 (m, 1H), 8.21 (s, 1H), 7.91 (d, J = 8.1 Hz, 1H), 7.63 (dd, J = 8.1, 10 1.2 Hz, 1H), 6.83 (m, 1H), 6.42 (m, 1H), 5.89 (m, 1H) , 4.77 (s, 2H), 4.68 (m, 1H), 4.41 (m, 1H), 3.87 (s, 3H), 2.98 (m, 1H), 2.40 ( m, 1H).

Example 44: 2- (2- (6-Cyano-1H-Acid

benzo [d] imidazola-1-yl) -9- ((R) -6,8-difluorochromano-4-yl) -8-oxo-8,9-dihydropurin-7-yl) acetic

To a solution of 2- (2- (6-cyano-1H-

benzo [d] imidazola-1-yl) -9 - ((- R) -6,8-difluorochromano-4-yl) -8-oxo

Methyl 8,9-dihydropurin-7-yl) acetate (4 mg) in MeOH (0.75 mL) and THF (0.25 mL) at 0 ° C was added a solution of lithium hydroxide (2 mg) in water (0.25 mL). The reaction was allowed to warm to room temperature and stirred for 2 h. After concentration under reduced pressure, the residue was diluted with water and acidified with 2 N HCl to pH = 4. The precipitated solid was filtered, washed with a small amount of cold water, treated with 1 N HCl in MeOH (0.5 mL) and concentrated in vacuo to afford 2 mg of the title compound. 1H-NMR (300 MHz, CDCl3) δ 9.04 (s, 1H), 8.75 (m, 1H), 8.46 (s, 1H), 7.87 (d, J = 8.4 Hz, 1H), 7.68 (dd, J = 8.4, 1.5 Hz, 1H), 6.92 (m, 1H), 6.70 (m, 1H), 5.97 (m, 1H), 4.77 (s, 2H), 4.63 (m, 1H), 4.44 (m, 1H), 2.91 (m, 1H), 2.42 (m, 1H). 10

15

20

Example 45: Synthesis of 2- (6-Fluoro-1H-benzo [d] imidazola-1-yl) -7- (piperidin-4-yl) -9- (tetrahydro-2H-pyran-4-yl) -7H-purine-8 (9H) -one -O.

^ 1)

HO-

N-Boc -> -

DIAD1 DCE Q-PPh3 2} H +

Resin-bound triphenylphosphine (264 mg, 2.15 mM / g, Argonaut) and tertiary butyl 4-hydroxypiperidine-1-carboxylate

6 (24 mg) was added to 2- (6-fluoro-1H-benzo [d] imidazole).

1-yl) -9- (tetrahydro-2H-pyran-4-yl) -7H-purine-8 (9H) -one in

dichloroethane (6 mL); The mixture was stirred for 10 min, when diisopropyl azodicarboxylate (64 µL) was added. After 18 h, the material was filtered, the resins were completely rinsed with ACN and MeOH and the combined organics were evaporated and purified by reverse phase HPLC (Boc with evaporative removed TFA) to obtain the title compound as a TFA salt. (16 mg). 1H-NMR (300 MHz, CDCl3 + 5% CD3OD) δ 9.0 (s, 1H), 8.6 (s, 1H), 8.3 (dd, 1H), 7.7 (dd, 1H) , 7.1 (td, 1H), 4.7-

4.4 (m, 2H), 4.2 (dd, 2H), 3.6 (t, 2H), 3.3 (br d, 2H), 2.9-2.7 (m, 4H), 2.4-2.2 (m, 4H), 2.9 (m, 1H), 1.9 (d, 2H), 1.8 (d, 2H); 19F δ -76, -117 ppm; MH + = 438; IR (CDCl3) 1726 (extension C = O).

\

2-1H-benzo [d] imidazol-1-yl) -9 - ((R) -furochromano-4-yl) -7-methyl-7H-purine-8 (9H) -one

This compound was synthesized by

method

described in example 38. 1H-NMR (300 MHz, CDCl3) δ 8.8 (s, 1H), 8.2 (s, 1H), 8.1 (m, 1H), 7.8 (m, 2H) , 7.3 (m, 2H), 7.0 (m, 1H), 6.7 (m, 2H), 5.9 (t, 1H), 4.7 (m, 1H), 4.4 ( (d, 1H), 3.6 (s, 3H), 2.9 (m, 1H), 2.4 (m, 1H). 10

2-1H-benzo [d] imidazol-1-yl) -9- (6-fuorochroman-4-yl) -7-methyl-7H-purine-8 (9H) -one

This compound was synthesized by the method described in example 38. 1 H NMR (300 MHz, CDCl 3): δ 8.80 (s, 1H), 8.19 (s, 1H), 7.96 (d, 1H), 7, 78 (d, 1H), 7.2-7.4 (m, 2H), 6.8-7.1 (m, 2H), 6.61 (dd, 1H), 5.88 (br t, 1H ), 4.54 (m, 1H), 4.32 (m, 1H), 3.56 (s, 3H), 2.84 (m, 1H), 2.30 (m, 1H).

XXj- °

15

2- (β-benzo [d] imidazola-1-yl) -9- ((R) -5,8-difluorochromano-4-yl) -7-methyl-7H-purine-8 (9H) -one

This compound was synthesized by the method described in example 38. 1 H NMR (300 MHz, CDCl 3): δ 8.7 (s, 1H), 8.2 (s, 1H), 8.0 (d, 1H), 7, 9 (d, 1H), 7.4 (m, 2H), 7.0 (td, 1H), 6.5 (td, 1H), 6.0 (t, 1H), 4.6 (m, 1H) ), 4.4 (m, 1H), 3.5 (s, 3H), 2.6 (m, 1H), 2.5 (m, 1H). 2- (β-benzo [d] imidazola-1-yl) -9 - ((R) -5,8-difluorochromano-4-yl) -7-ethyl-7H-purine-8 (9H) -one

This compound was synthesized by the method described in example 38 using iodine ethane. 1H NMR (300 MHz, CDCl3): δ 8.8 (s, 1H), 8.2 (s, 1H), 8.0 (d, 1H), 7.9 (d, 1H), 7.4 ( m, 2H),

7.0 (td, 1H), 6.5 (td, 1H), 6.0 (t, 1H), 4.6 (m, 1H), 4.4 (m, 1H), 4.0 (m 2H), 2.6 (m, 1H), 2.5 (m, 1H), 1.5 (t, 3H).

F

2- (β-benzo [d] imidazol-1-yl) -9- ((R) -5,8-difluorochromano-4-yl) -7-propyl-7H-purine-8 (9H) -one

This compound was synthesized by the method

described in example 38 using iodopropane. 1H NMR (300 MHz, CDCl3): δ 9.8 (sr IH), 8.2 (s, 1H), 8.0 (d, 1H), 7.9 (d, 1H), 7.4 (m , 2H),

7.0 (td, 1H), 6.5 (td, 1H), 6.0 (t, 1H), 4.6 (m, 1H), 4.4 (m, 1H), 4.0 (m 2H), 2.6 (m, 1H), 2.5 (m, 1H), 1.9 (m, 2H), 1.0 (t, 3H).

F

9- ((R) -5,8-difluorochromano-4-yl) -2- (6-fluoro-1H-benzo [d] imidazola-1-yl) -7-methyl-7H-purine-8 (9H) -ona

This compound was synthesized by the method described in example 38. 1H-NMR (300 MHz, CDCl3) δ 8.7 (s, 1H), 8.0 (m, 1H), 7.7 (dd, 1H), 7, 1 (m, 2H), 6.5 (td, 1H), 5.9 (dd, 1H),

4.6 (m, 1H), 4.5 (m, 1H), 3.5 (s, 3H), 2.6 (m, 1H), 2.5 (m, 1H). F

9- ((R) -5,8-difluorochromano-4-yl) -2- (6-fluoro-1H-benzo [d] imidazola-1-yl) -7- (2-methoxy-ethyl) -7H- purine-8 (9H) -one

This compound was synthesized by the method described in example 39. 1H-NMR (300 MHz, CDCl3) δ 8.7 (s, 1H), 8.4

9- ((R) -5,8-difluorochroman-4-yl) -7- (2- (dimethylamine) ethyl) -2- (6-fluoro-1H-benzo [d] imidazola-1-yl) Purine-8 (9H) -one

This compound was synthesized by the method described in example 41. 1H-NMR (300 MHz, CDCl3) δ 9.2 (s, 1H), 8.5 (s, 1H), 7.9 (m, 1H), 7, 8 (dd, 1H), 7.2 (td, 1H), 7.1 (td, 1H),

6.5 (td, 1H), 5.9 (t, 1H), 4.6 (m, 1H), 4.5 (m, 3H), 3.6 (m, 2H), 3.0 (s , 6H), 2.7 (m, 1H), 2.5 (m, 1H).

(s, 1H)., 8.0 (m, 1H), 7.8 (dd, 1H), 7.2-7.0 (m, 2H), 6.5 (td, 1H)

6.0 (t, 1H), 4.6 (m, 1H), 4.5 (m, 1H), 4.1 (t, 2H), 3.7 (t, 2H)

3.4 (s, 3H), 2.7 (m, 1H), 2.4 (m, 1H).

F 2- (6-Chloro-1H-benzo [d] imidazole-1-yl) -9- ((R) -8-fluorochroman-4-yl) -7-methyl-7H-purine-8 (9H) - one

This compound was synthesized by the method described in example 38. 1H-NMR (300 MHz, CDCl3) δ 8.7 (s, 1H), 8.3 (m, 1H), 8.2 (s, 1H), 7, 7 (d, 1H), 7.3 (dd, 1H), 7.0 (td, 1H),

6.8-6.6 (m, 2H), 5.9 (dd, 1H), 4.7 (m, 1H), 4.5 (td, 1H), 3.6 (s, 3H), 3 .0 (m, 1H), 2.4 (m, 1H).

NC \

3- (7-methyl-8-oxo-9- (tetrahydro-2H-pyran-4-yl) -8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-5-carbonitrile

This compound was synthesized by the method described in example 38. 1H-NMR (300 MHz, CDCl3) δ 9.37 (s, 1H), 8.90. (m, 1H), 8.58 (s, 1H), 8.00 (d, J = 8.4 Hz, 1H), 7.79 (dd, J = 8.4, 1.5 Hz, 1H) , 4.63 (m, 1H), 4.03 (m, 2H), 3.51 (m, 2H), 3.44

NC I

3- (9-chromano-4-yl-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzoimidazole-5-carbonitrile

This compound was synthesized by the method described in example 38. 1H-NMR (300 MHz, CDCl3) δ 8.8 (s, 1H), 8.5 (s, 1H), 8.2 (s, 1H), 7, 8 (d, 1H), 7.6 (d, 1H), 7.2 (m, 2H), 6.9 (d, 1H), 6.8 (d, 1H), 5.9 (t, 1H) ), 4.6 (m, 1H), 4.4 (m, 1H), 3.6 (s, 3H), 2.8 (m, 1H), 2.4 (m, 1H). NC I

3- [9- (8-Fluoro) -chroman-4-yl) -7-methyl-8-oxo

8,9-dihydro-7H-purin-2-yl] -3H-benzoimidazole-5-carbonitrile

This compound was synthesized by the method described in example 38. 1H-NMR (300 MHz, CDCl3) δ 8.8 (d, 2H), 8.2 (s, 1H), 7.8 (d, 1H), 7.6 (d, 1H), 7.0 ( t, 1H), 6.6 (m, 2H), 5.9 (t, 1H), 4.6 (m, 1H), 4.4 (m, 1H), 3.6 (s, 3H), 2.8 (m, 1H), 2.4 (m, 1H).

3- (9 - ((R) -6-fluoro) chroman-4-yl) -7-methyl-8-oxo

8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-5-carbonitrile

This compound was synthesized by the method described in Example 38. 1 H NMR (300 MHz, CDCl 3): δ 8.90 (s, 1H), 8.52 (s, 1H), 8.21 (s, 1H), 7, 84 (d, 1H), 7.59 (d, 1H), 7.1-7.2 (m, 1H), 6.9-7.0 (m, 1H), 6.59 (dd, 1H) , 5.89 (br t, 1H), 4.5-4.6 (m, 1H), 4.34 (br t, 1H), 3.59 (s, 3H), 2.8-2.9 (m, 1H), 2.3-2.4 (m, 1H). 3- (9- ((R) -7-fluorochroman-4-yl) -7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-5-carbonitrile

This compound was synthesized by the method described in example 38. 1H NMR (300 MHz, CDCl3): δ 8.90 (s, 1H), δ

15

8.62 (s, 1Η), 8.19 (s, 1H), 7.84 (d, 1Η), 7.59 (d, 1Η), 6.8-7.0 (m, 2H), 6.5-6.6 (m, 1H), 5.86 (br t, 1H), 4.5-4.6 (m, 1H), 4.36 (br t, 1H), 3.59 ( s, 3H), 2.8-2.9 (m, 1H), 2.3-2.4 (m, 1H).

AT THE \

3- (9- ((R) -6,8-difluorochromano-4-yl) -7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzo [ d] imidazole-5-carbonitrile

This compound was synthesized by the method described in example 38. 1H-NMR (300 MHz, CDCl3) δ 8.9 (s, 1H), 8.8 (s, 1H), 8.2 (s, 1H), 7, 9 (d, 1H), 7.6 (dd, 1H), 6.8 (td, 1H), 6.4 (dd, 1H), 5.9 (dd, 1H), 4.7 (m, 1H) ), 4.4 (td, 1H), 3.6 (s, 3H),

3.0 (m, 1H), 2.4 (m, 1H).

ϊΛ- °

3- (9- (5,8-difluorochroman-4-yl) -7-methyl-8-oxo

8,9-dihydro-7H-purin-2-yl) -3H-benzoimidazole-5-carbonitrile

This compound was synthesized by the method described in example 38. 1H-NMR (300 MHz, CDCl3) δ 8.8 (s, 1H), 8.7 (s, 1H), 8.2 (s, 1H), 7, 8 (d, 1H), 7.6 (d, 1H), 7.0 (m, 1H), 6.5 (m, 1H), 5.9 (t, 1H), 4.5 (m, 2H) ), 3.5 (s, 3H), 2.5 (m, 2H). 3- [7-Ethyl-9- (8-fluorochroman-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-yl] -3H-benzoimidazole-5-carbonitrile

This compound was synthesized by the method

described in example 38 using iodoethane. 1H-NMR (300 MHz, CDCl3) δ 8.8 (s, 2H), 8.2 (s, 1H), 7.9 (d, 1H), 7.6 (d, 1H), 7.0 (t, 1H), 6.7 (m, 2H), 5.9 (t, 1H), 4.6 (m, 1H), 4.4 (m, 1H), 4.0 (q, 2H) 2.9 (m, 1H), 2.4 (m, 1H), 1.4 (t, 3H). 3- (7-ethyl-9 - ((R) -7-fluorochroman-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-5-carbonitrile

This compound was synthesized by the method

described in example 38 using iodoethane. 1H NMR (300 MHz, CDCl3) δ 8.88 (s, 1H), 8.62 (s, 1H), 8.21 (s, 1H), 7.84 (d, 1H), 7.59 (d , 1H), 6.8-7.0 (m, 2H), 6-4-6.6 (m, 1H), 5.86 (br t, 1H), 4.5-4.6 (m, 4.36 (br t, 1H), 4.0-4.1 (m, 2H), 2.8-2.9 (m, 1H), 2.3-2.4 (m, 1H) ), 1.48 (t, 3H).

NO 3- (9- ((R) -6,8-difluorochromano-4-yl) -7-ethyl-8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzo [d] imidazole-5-carbonitrile

This compound was synthesized by the method described in example 38 using iodine ethane. 1H-NMR (300 MHz, CDCl3) δ

8.9 (s, 1H), 8.8 (s, 1H), 8.2 (s, 1H), 7.9 (d, 1H), 7.6 (dd, 1H),

6.8 (td, 1H), 6.4 (dd, 1H), 5.9 (dd, 1H), 4.7 (m, 1H), 4.4 (td, 1H), 4.1 (q , 2H), 3.0 (m, 1H), 2.4 (m, 1H), 1.5 (t, 3H).

10

15

20

3- (9- (5,8-difluorochroman-4-yl) -7-ethyl-8-oxo

8,9-dihydro-7H-purin-2-yl) -3H-benzoimidazole-5-carbonitrile

This compound was synthesized by the method described in example 38 using iodine ethane. 1H-NMR (300 MHz, CDCl3) δ

8.8 (s, 1H), 8.7 (s, 1H), 8.2 (s, 1H), 7.9 (d, 1H), 7.6 (d, 1H),

7.0 (m, 1H), 6.4 (m, 1H), 5.9 (t, 1H), 4.6 (m, 1H), 4.4 (m, 1H),

4.0 (q, 2H), 2.5 (m, 2H), 1.4 (t, 3H).

NC

3- [9- (5,8-Difluorochroman-4-yl) -7-isobutyl-8-oxo-8,9-dihydro-7H-purin-2-yl] -3H-benzoimidazole-5-one carbonitrile

This compound was synthesized by the method described in example 38 using iodide isobutyl. 1H-NMR (300 MHz, CDCl3) δ 8.9 (s, 1H), 8.8 (s, 1H), 8.2 (s, 1H), 7.9 (d, 1H), 7.6 d , 1H), 7.1 (m, 1H), 6.5 (m, 1H), 5.9 (t, i --- I (m, 1H), 4.4 m, 1H), 3.8 (q, 2H), 2.7 (m, 1H), 2.5 (m, 1H), 2.3 (q, 1H), 1.0 s, 6H). 3- (7-benzyl-8-oxo-9- (tetrahydro-2H-pyran-4-5-yl) -8,9-dihydro-7H-purin-2-yl) -3H-benzo [d ] imidazole-5-carbonitrile

This compound was synthesized by the method described in example 38 using benzyl bromide. 1H NMR (300 MHz, CDCl3): δ 9.30 (s, 1H), 9.01 (m, 1H), 8.23 (s, 1H), 7.89 (m, 1H), 7.66 ( dd, J = 8.1, 1.5 Hz, 1H), 7.40-7.32 (m, 5H), 5.16 (s, 2H), 4.75 (m, 1H), 4.18 (m, 2H), 3.63 (m, 2H), 2.81 (m, 2H), 1.90 (m, 2H).

NC \ OH

3- (9- (6,8-difluorochroman-4-yl) -7- (3-hydroxypropyl) -8-oxo-8,9-dihydro-7H-purin-2-yl) -3H-benzo [ d] imidazole-5-carbonitrile

This compound was synthesized by the method described in example 40 using 3-iodopropanol. 1H NMR (300 MHz, CDCl3): δ 8.92 (s, 1H), 8.77 (m, 1H), 8.38 (s, 1H), 7.90 (d, J = 8.4 Hz, 7.62 (dd, J = 8.4, 1.5 Hz, 1H), 6.83 (m, 1H), 6.38 (m, 1H), 5.88 (m, 1H), 4.67 (m, 1H), 4.41 (m, 1H), 4.19 (t, J = 6.5 Hz, 2Η), 3.74 (t, J = 5.6 Hz, 2Η), 2.93 (m, 1Η), 2.40 (m, 1Η),

2.08 (m, 2Η).

'νΛ

3- (7- (2- (diethylamine) ethyl) -9 - ((R) -T-fluorochromano-4-yl) -8-oxo-8,9-dihydro-7H-purin-2-yl ) -3H-benzo [d] imidazole-5-carbonitrile

This compound was synthesized by the method described in example 41 using 2-chloro-N, N-diethylethylamine hydrochloride. 1H NMR (300 MHz, CDCl3): δ 8.91 (s, 1H), 8.64 (s, 1H), 8.28 (s, 1H), 7.84 (d, 1H), 7.59 ( d, 1H), 6.8-7.0 (m, 2H), 106.6-6.6 (m, 1H), 5.86 (br t, 1H), 4.5-4.7 ( m, 1H), 4.3-4.4 (m, 1H), 4.0-4.1 (m, 2H), 2.8-3.0 (m, 3H), 2.58 (q, 4H), 2-3-4 (m, 1H), 0.96 (t, 6H).

Jak3 Kinase Assay

Human Jak3 DNA was amplified by PCR 15 (polymerase chain reaction). A fragment encoding the Jak3 catalytic domain (508aa through 1124aa) was ligated with GST (glutathione S-transferase) at the 5 'end. This fused GST-Jak3 DNA fragment was cloned into the EcoRI site of donor plasmid pFastBac 1 (Life Technologies # 10359-016). Insect cell transformation, transposition and transfection (Sf9) were performed according to the manufacturing instructions. Recombinant GST-Jak3 containing cell lysate was used in the kinase assay. Anti-GST antibodies (10 μg / mL, Sigma # G1417) were placed in a 384-well plate at 4 ° C overnight. GST-Jak3-containing cell lysate (1: 100 dilution) was added to the anti-GST-coated plates and GST-Jak3 was captured by immobilized anti-GST antibodies. Test compounds and substrate mixture (50 mM HEPES, pH = 7, 0.5 mM Na3VO4, 25 mM MgCl2, 1 mM DTT, 0.005% BSA, 1 μπι ATP and 4.5 μg / ml of poly-Glu biotinyl, Ala, Tir) was added to the plate to initiate the reaction. After a 60 min incubation, the reaction was stopped by 4 mM EDTA and poly-Glu biotinyl Ala Tir phosphorylation was detected using 17 μg / ml 5 Cy5-streptavidin (Amersham, # PA92005) and 2, 7 μg / ml Europium-conjugated anti-phosphotyrosine antibodies (PerkinElmer # AD0069) using homogeneous time-resolved fluorescence (FTRH) technology.

Jak3 Cell Test

The pre-B F7 lymphocyte cell line of

rat was used for the Jak3 cell assay. Human IL-2 cRNA cDNA is stably expressed in F7 cells (Kawahara et al., 1995). F7 cells were maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum plus IL-3. Cells 15 (30,000 cells / well) in serum free medium were plated in 96 well plates for the cell proliferation assay. Test compounds were added to the cells, followed by the addition of IL-2 (20 ng / ml final). Following a 24 hr incubation the number of viable cells was determined by the CellTiter-Glo Luminescent Cell Viability Assay (Promega, # G7573) according to the manufacturer's instructions.

Results of representative species tests are shown below. The compounds in Table 1 exhibited IC50 less than 100 nM. The compounds in Table 1 exhibited IC50 between 101 nM and 1 μπι. Synthesis Example

Reference N0

504

505

CN

506 Synthesis Example

Reference N0

Example

40

522

523

Example

39

524

OCH3 HO Table I

IL-2-induced IFN-γ production in rat

IL-2 administration leads to increased serum IFN-γ in the rat due to cytokine AN (natural killer) secretion [Thornton S., Kuhn KA, Finkelman FD and Hirsch R., "AN cells secrete high levels of IFN-γ in response to In vivo administration of IL-2 "(NK cells secreting high levels of IFN-γ in response to in vivo administration of IL-2), Eur. J. Immunol2001 31: 3355-3360]. The experiment was performed essentially according to the protocol in Thornton et al. and test compounds were administered to determine the level of inhibition achieved. In summary, female BALB / c mice were fasted for 12-18 hours prior to a study but had free access to water at all times. Test compounds were administered by gavage one hour before intraperitoneal injection of IL-2 and capture antibodies. At the end of the studies, rats were sacrificed by carbon dioxide inhalation, terminal blood samples were collected by cardiac puncture and serum was generated. Serum was stored frozen until assayed for IFN-γ as described by the equipment manufacturer (BD Farmingen®, San Diego, CA).

Using the above method, compounds of Experimental Examples 16, 28 and Examples 501, 504 and 505 of Table 1 at mg / kg have been shown to inhibit IL-2-induced IFN-γ production by more than 40% in vivo. , in the rat. A reference compound, CP690550, exhibited 96% inhibition at 30 mg / kg in this screen.

The 7-substituted purinones exhibited increased selectivity for Jak3 compared to their unsubstituted counterparts.

The Aurora A kinase assay was performed using

a fluorescence polarization format. A 100 mM solution of fluorescence-labeled FAM PKAtide (Molecular Devices), the substrate for Aurora A (Upstate Biotechnologies), was incubated with Aurora A (80 ng / ml) and 30 mM ATP at room temperature 35 for 1 hour. in the presence of an appropriate concentration of a test inhibitor. The reaction was terminated by the addition of a mixture of IMAP Progressive Binding Reagent according to the manufacturer's instructions (Molecular Devices). The bias signal was detected using Aquest (Molecular Devices).

below, down, beneath, underneath, downwards, downhill. On the left are the unsubstituted compounds and their Jak3 rate for Aurora A inhibition, given as "x-fold" selectivity. On the right are 7-substituted counterparts along with their selectivity. All IC50 for Jak3 are below 100 nM.

Some comparative examples are shown.

NOT REPLACED 7

7-REPLACED

112 times

H

1

14 times

58 times

CH3

NC

^ N times H

47 times

58 times ΝΑΟ-7-REPLACED

7-REPLACED

13 times

H

/

NC

already

1S2 times CHs p r-O Í

'■ The r ~ \ J

rVNYNy N F

\ = / I sl> = o

ISlC

Et

I40 times .o

Γ v = <

NC

N

178 times

CPyto him

f. >

/ NC

V

11 times

OH

\ _ / ~~ OH

200 times ΝΑΟ-7-REPLACED

7-StJBST ITUIDOS

238 times GH3

(Hv F

V = '> = 0

NC!

Et

123 times

NC

f

V ^ N (CH3) 2 724 times

While the above invention has been described in some detail for illustration purposes, it will be readily apparent to those skilled in the art that changes and modifications may be made without departing from the scope of the invention described herein.

Claims (44)

1. "7-SUBSTITUTED PURINE DERIVATIVES FOR IMMUNOSUPPRESSION", characterized in that they comprise a compound of formula III: where Q1 and Q2 are independently selected from the group consisting of in CXi, CX2 and nitrogen, where Q1 and Q2 are not both nitrogen; Q3 is N or CH; X 1 and X 2 are independently selected from the group consisting of hydrogen, (C 1 -C 6) alkyl, cyano, halo, (C 1 -C 6) alkyl halo, hydroxyl, (C 1 -C 6) alkoxy; (C 1 -C 6) alkoxy and nitro; is selected from the group consisting of hydrogen and (C1-C6) alkyl; y is zero or an integer selected from 1, 2 and 3; R2 and R3 are independently selected for each occurrence of (CR2R3) from the group consisting of hydrogen and (C1-C6) alkyl R4 is selected from a group consisting of alkyl, heterocyclyl, aryl, heteroaryl, substituted alkyl, substituted heterocyclyl, substituted aryl, substituted heteroaryl and R5 is selected from the group consisting of alkyl, heterocyclyl, substituted heterocyclyl and C1-C6 alkyl, wherein (a) one or two CH2 are substituted by a group selected from NH and N (alkyl); (b) one or two CH2 are replaced by O; (c) one or two CH 2 are replaced by (C = O); (d) two CH2 are replaced by CH = CH or C = C; or (e) any chemically stable combination of (a), (b) (c) and (d); and wherein from zero to three hydrogens are substituted by a substituent chosen from: (a) halogen, hydroxy, cyano, lower alkylsulfonyl, lower alkylsulfonyloxy, amino, lower alkylamino, lower dialkylamino, alkoxyamino, sulfonylamino, acylamino, arylamino, alkoxy bottom; (b) heterocyclyl and heterocyclyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl; (c) phenyl and phenyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl, acylamino, cyano, carboxy, alkoxycarbonyl, haloalkyl and heterocyclyl; and (d) heteroaryl and heteroaryl substituted with one to three substituents chosen from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 1, characterized in that they comprise a compound of the formula: "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 2, characterized in that they comprise a compound of the formula: <formula> formula see original document page 126 </formula> "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 2, characterized in that they comprise a compound of the formula: <formula> formula see original document page 126 </formula> "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 1, characterized in that they comprise a compound of the formula: <formula> formula see original document page 126 </formula> "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 5, characterized in that they comprise a compound of the formula: <formula> formula see original document page 127 </formula> 7. "SUBSTITUTED PURINE DERIVATIVES" according to claim 5, characterized in that they comprise a compound of the formula: <formula> formula see original document page 127 </formula> "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 1, characterized in that they comprise a compound of the formula: <formula> formula see original document page 127 </formula> 9. "SUBSTITUTED PURINE DERIVATIVES" according to claim 8, characterized in that they comprise a compound of the formula: <formula> formula see original document page 128 </formula> "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 8, characterized in that they comprise a compound of the formula: <formula> formula see original document page 128 </formula> "7-SUBSTITUTED PURINE DERIVATIVES" according to claims 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, characterized in that X1 and X2 are independently selected from hydrogen, cyano, chlorine, fluorine, methyl, trifluoromethyl and trifluoromethoxy. "7-SUBSTITUTED PURINE DERIVATIVES" according to claims 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, characterized in that R1 is H. "7-SUBSTITUTED PURINE DERIVATIVES" according to claims 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, characterized in that y is 1 or 2 and R2 and R3 are hydrogen or methyl. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 13, characterized in that R4 is selected from phenyl, quinoline, pyridine, pyrazine and their substituted counterparts. "7-SUBSTITUTED PURINE DERIVATIVES" according to claims 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, characterized in that y is zero. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 15, characterized in that R4 is selected from cyclopentyl, cyclohexyl, phenyl, indane, tetraline, piperidine, oxepane, benzoxepane dihydrocyclopentapyridine, tetramide. hydropyran, tetrahydrofuran, tetrahydro-indole, isoquinoline, tetrahydro-isoquinoline, quinoline, tetrahydroquinoline, chroman, pyridine, pyrimidine, dihydropyran, dihydrobenzofuran, tetrahydrobenzofuran, tetrahydrobenzothiophene, furane dihydropyran [2,3-b] pyridine, tetrahydroquinoxaline, tetrahydrothiopyran (thian), thiochroman (dihydrobenzothiine) and their substituted counterparts. "7-SUBSTITUTED PtTRINE DERIVATIVES" according to claims 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, characterized in that (a) y is zero and R4 is selected from cyclohexyl, tetraline, indane, oxepane, benzoxepane, dihydrocyclopentapyridine, tetrahydropyran, tetrahydroquinoline, chroman, dihydrobenzofuran, tetrahydrobenzofuran, dihydropyran [2,3-b] pyridine and tetrahydro hydroquinoxaline, each optionally substituted with hydroxy, oxo, or halogen; or (b) y is 1 or 2, R 2 and R 3 are hydrogen or methyl and R 4 is selected from phenyl, pyridine and pyrazine, each optionally substituted with halogen. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 17, characterized in that y is 0 and R4 is chosen from chroman-4-yl; 3,4-dihydronaphthalene-1 (2H) -one-4-yl; 2,3-dihydro-inden-1-one-4-yl and its fluorine substituted counterparts. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 18, characterized in that R4 is chroman-4-yl and the carbon at position 4 of chroman is of the (R) configuration. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 17, characterized in that y is 0 and R4 is: wherein W is CH2, C = O or O; p is 1, 2 or 3; A is a six-membered heteroaromatic ring containing 1 or 2 nitrogen or a benzene ring optionally substituted with one or two fluorines; and the wavy line is the purinone attachment point. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 20, characterized in that the carbon labeled with an asterisk <formula> formula </b> is of the (R) configuration. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 17, characterized in that y is I and R4 is selected from difluorophenyl, fluorophenyl, chlorophenyl, chlorofluorophenyl, pyridin-3-yl and pyrazine-3-yl . "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 17, characterized in that y is zero and R 4 is selected from tetrahydropyran-4-yl, 4-hydroxycyclohexyl, 4-oxocycle -hexyl and oxepan-4-yl. 24. "SUBSTITUTED PURINE DERIVATIVES" according to claim 10, characterized in that X1 and X2 are independently selected from hydrogen, cyano, chlorine and fluorine and R1 is H. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 24, characterized in that (a) y is zero and R 4 is selected from cyclohexyl, tetraline, indane, oxepane, benzoxepane, dihydrocyclopentapyridine, tetrahydropyran, tetrahydroquinoline, chroman, dihydrobenzofuran, tetrahydrobenzofuran, dihydropyran [2,3-b] pyridine and tetrahydroquinoxaline, each optionally substituted with hydroxy, oxo, or halogen; or (b) y is 1 or 2, R 2 and R 3 are hydrogen or methyl and R 4 is selected from phenyl, pyridine and pyrazine, each optionally substituted with halogen. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 25, characterized in that R5 is C1 -C6 alkyl, wherein (a) one or two CH2 are substituted by a group selected from NH and N ( alkyl); (b) one or two CH2 are replaced by O; (c) one or two CH 2 are replaced by (C = O); (d) two CH2 are replaced by CH = CH or C = C; or (e) any chemically stable combination of (a), (b) (c) and (d); and wherein from zero to three hydrogens are substituted by a substituent chosen from: (a) halogen, hydroxy, cyano, lower alkylsulfonyl, lower alkylsulfonyloxy, amino, lower alkylamino, lower alkylamino, alkoxyamino, sulfonylamino, acylamino, arylamino lower alkoxy; (b) heterocyclyl and heterocyclyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl; (c) phenyl and phenyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl, acylamino, cyano, carboxy, alkoxycarbonyl, haloalkyl and heterocyclyl; and (d) heteroaryl and heteroaryl substituted with one to three substituents chosen from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 26, characterized in that y is 0 and R4 is wherein the carbon labeled with an asterisk is of configuration (R); W is CH 2, C = O or 0; p is 1, 2 or 3; A is a six membered heteroaromatic ring containing 1 or 2 nitrogen or a benzene ring optionally substituted with one or two fluorines; and the wavy line is the purinone attachment point. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 27, characterized in that X 1 is hydrogen, X 2 is a 6-position substituent on benzimidazole and X 2 is chosen from hydrogen, fluorine and cyano. "7-SUBSTITUTED PURINE DERIVATIVES" according to claims 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, characterized in that R5 is C1 -C6 alkyl or C1-6. C6 fluoroalkyl. 30, "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 29, characterized in that y is 0 and R4 is wherein the carbon labeled with an asterisk is of configuration (R); W is CH 2, C = O or 0; p is I, 2 or 3; A is a six membered heteroaromatic ring containing 1 or 2 nitrogen or a benzene ring optionally substituted with one or two fluorines; and the wavy line is the purinone attachment point. "7-SUBSTITUTED PURINE DERIVATIVES" according to claims 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, characterized in that R5 is C1 -C6 alkyl and wherein from zero to three hydrogens are substituted by a substituent chosen from: hydroxy, carboxy, cyano, lower alkylsulfonyl, lower alkylsulfonyloxy, amino, lower alkylamino, lower alkylamino, alkoxyamino, sulfonylamino, acylamino, arylamino and lower alkoxy. 32. "SUBSTITUTED PURINE DERIVATIVES" according to claim 31, characterized in that y is 0 and R4 is wherein the carbon labeled with an asterisk is from configuration (R); W is CH 2, C = O or 0; p is 1, 2 or 3; A is a six membered heteroaromatic ring containing 1 or 2 nitrogen or a benzene ring optionally substituted with one or two fluorines; and the wavy line is the purinone attachment point. 33. "SUBSTITUTED PURINE DERIVATIVES" according to claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 or 32, characterized in that they comprise a pharmaceutical composition including a pharmaceutically acceptable carrier and a therapeutically effective amount of at least at least one compound according to any one of claims 1 to 32. 34. "7-SUBSTITUTED PURINE DERIVATIVES FOR IMMUNOSUPRESSION", characterized in that they comprise a method for treating a disorder that is dependent on inhibition of Janus kinase 3, which comprises administering to an individual in need of such treatment an amount therapeutically effective composition of a compound according to formula III: wherein Q1 and Q2 are independently selected from the group consisting of CX1, CX2 and nitrogen, where Q1 and Q2 are not. they are both nitrogen; Q3 is N or CH; X1 and X2 are independently selected from the group consisting of hydrogen, (C1-C6) alkyl, cyano, halo, (C1-C6) alkyl, hydroxyl, (C1-C6) alkoxy; (C1-C6) alkoxy and nitro; R1 is selected from the group consisting of hydrogen and (C1-C6) alkyl; y is zero or an integer selected from 1, 2 and 3; R2 and R3 are independently selected for each occurrence of (CR2R3) from the group consisting of hydrogen and (C1-C6) alkyl R4 is selected from a group consisting of alkyl, heterocyclyl, aryl, heteroaryl, substituted alkyl, substituted heterocyclyl, substituted aryl, substituted heteroaryl and R5 is selected from the group consisting of C 1 -C 6 alkyl, heterocyclyl, substituted heterocyclyl, wherein (a) one or two CH 2 are substituted by a group selected from NH and N (alkyl); (b) one or two CH 2 are substituted by 0; (c) one or two CH 2 are replaced by (C = O); (d) two CH2 are replaced by CH = CH or C = C; or (e) any chemically stable combination of (a), (b) (c) and (d); and wherein from zero to three hydrogens are substituted by a substituent chosen from: (a) halogen, hydroxy, cyano, lower alkylsulfonyl, lower alkylsulfonyloxy, amino, lower alkylamino, lower dialkylamino, alkoxyamino, sulfonylamino, acylamino, arylamino, alkoxy bottom; (b) heterocyclyl and heterocyclyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl; (c) phenyl and phenyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl, acylamino, cyano, carboxy, alkoxycarbonyl, haloalkyl and heterocyclyl; and (d) heteroaryl and heteroaryl substituted with one to three substituents chosen from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl. 35. "7-SUBSTITUTED PURINE DERIVATIVES FOR IMMUNOSUPRESSION", characterized in that they comprise a method for treating a disorder that is dependent on inhibition of Janus kinase 3, which comprises administering to an individual in need of such treatment an amount A therapeutically effective compound of a compound according to formula III: wherein Q1 and Q2 are independently selected from the group consisting of CXi, CX2 and nitrogen, where Qi and Q2 are not. they are both nitrogen; Q3 is N or CH; X1 and X2 are independently selected from the group consisting of hydrogen, cyano, chlorine and fluorine; R1 is hydrogen; y is zero, or an integer selected from 1, 2 and 3; R2 and R3 are independently selected for each occurrence of (CR2R3) from the group consisting of hydrogen and (C1-C6) alkyl; R4 is selected from a group consisting of alkyl, heterocyclyl, aryl, heteroaryl, substituted alkyl, substituted heterocyclyl, substituted aryl, substituted heteroaryl; and R5 is selected from the group consisting of alkyl, heterocyclyl, substituted heterocyclyl and C1-C6 alkyl, wherein (a) one or two CH2 are substituted by a group selected from NH and N (alkyl); (b) one or two CH2 are replaced by O; (c) one or two CH 2 are replaced by (C = O); (d) two CH2 are replaced by CH = CH or C = C; or (e) any chemically stable combination of (a), (b) (c) and (d); and wherein from zero to three hydrogens are substituted by a substituent chosen from: (a) halogen, hydroxy, cyano, lower alkylsulfonyl, lower alkylsulfonyloxy, amino, lower alkylamino, lower dialkylamino, alkoxyamino, sulfonylamino, acylamino, arylamino, alkoxy bottom; (b) heterocyclyl and heterocyclyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl; (c) phenyl and phenyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl, acylamino, cyano, carboxy, alkoxycarbonyl, haloalkyl and heterocyclyl; and (d) heteroaryl and heteroaryl substituted with one to three substituents chosen from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl. A "7-SUBSTITUTED PURINE DERIVATIVE" according to claim 34 or 35, characterized in that said disorder is selected from an autoimmune disease, an inflammatory disease, a mast cell-mediated disease, a hematological malignancy and organ transplant rejection. 37. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 36, characterized in that said disorder is bone marrow transplant rejection. 38. "SUBSTITUTED PURINE DERIVATIVES" according to claim 36, characterized in that said hematological malignancy is selected from leukemia and lymphoma. 39. "SUBSTITUTED PURINE DERIVATIVES" according to claim 36, characterized in that said disorder is asthma. 40. "SUBSTITUTED PURINE DERIVATIVES" according to claim 36, characterized in that said autoimmune disease is selected from a specific autoimmune disease, to a specific organ and to a non-organ. 41. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 36, characterized in that said disorder is keratoconjunctivitis sicca. 42. "7-SUBSTITUTED PURINE DERIVATIVES" according to claim 36, characterized in that said hematologic malignancy is chronic myelogenous leukemia. A "7-SUBSTITUTED PURINE DERIVATIVE" according to claim 34 or 35, characterized in that said disorder is selected from a leukemic form of cutaneous T-cell lymphoma and acute lymphoblastic leukemia. 44. "7-SUBSTITUTED PURINE DERIVATIVES FOR SUPREME SUNS" are characterized by the fact that they comprise a method for treating a selected disorder of an autoimmune disease, an inflammatory disease, a mast cell-mediated disease, a hematological malignancy and organ transplant rejection, comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound according to formula III wherein Qi and Q2 are independently selected. from the group consisting of CXi, CX2 and nitrogen; wherein Qi and Q2 are not both nitrogen; Q3 is N or CH; X1 and X2 are independently selected from the group consisting of hydrogen, (C1-C6) alkyl, cyano, halo, (C1-C6) alkyl, hydroxyl, (C1-C6) alkoxy; halo (C1-C6) alkoxy and nitro; R1 is selected from the group consisting of hydrogen and (C1-C6) alkyl; y is zero or an integer selected from 1, 2 and 3; R2 and R3 are independently selected for each occurrence of (CR2R3) from the group consisting of hydrogen and (C1-C6) alkyl; R4 is selected from a group consisting of alkyl, heterocyclyl, aryl, heteroaryl, substituted alkyl, substituted heterocyclyl, substituted aryl, substituted heteroaryl; and R5 is selected from the group consisting of alkyl, heterocyclyl, substituted heterocyclyl and C1-C6 alkyl, wherein (a) one or two CH2 are substituted by a group selected from NH and N (alkyl); (b) one or two CH2 are replaced by O; (c) one or two CH 2 are replaced by (C = O); (d) two CH2 are replaced by CH = CH or C = C; or (e) any chemically stable combination of (a), (b) (c) and (d); and wherein from zero to three hydrogens are substituted by a substituent chosen from: (a) halogen, hydroxy, cyano, lower alkylsulfonyl, lower alkylsulfonyloxy, amino, lower alkylamino, lower dialkylamino, alkoxyamino, sulfonylamino, acylamino, arylamino, alkoxy bottom; (b) heterocyclyl and heterocyclyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl; (c) phenyl and phenyl substituted with one to three substituents selected from halogen, hydroxy, alkoxy, alkyl, acylamino, cyano, carboxy, alkoxycarbonyl, haloalkyl and heterocyclyl; and (d) heteroaryl and heteroaryl substituted with one to three substituents chosen from halogen, hydroxy, alkoxy, alkyl and alkoxycarbonyl.