Biaryl substituted purine derivatives as potent antiproliferative agents

The compounds of the present invention are 2,6,9-trisubstituted purine derivatives which are inhibitors of cyclin/cdk complexes. The compounds of the current invention also are potent inhibitors of human cellular proliferation. As such, the compounds of the present invention constitute pharmaceutical compositions with a pharmaceutically acceptable carrier. Such compounds are useful in treating a disorder mediated by elevated levels of cell proliferation in a mammal compared to a healthy mammal by administering to such mammal an effective amount of the compound. Examples of the compounds of the present invention are represented by the following chemical structures: wherein:

FIELD OF THE INVENTION

The present invention relates to compounds that are shown to be potent cyclin/cyclin dependent kinase (cdk) inhibitors. Compounds with these properties are shown to be potent inhibitors of cell growth and proliferation. Such compounds can be used to treat the following conditions in mammals: rheumatoid arthritis, lupus, type 1 diabetes, multiple sclerosis, cancer, restenosis, gout and other proliferative diseases involving elevated levels of proliferation compared to healthy mammals. Compounds of the present invention which are biaryl substituted purine derivatives are shown to be potent antiproliferative agents against a number of human transformed cell lines, and also inhibitors of human cyclin/cdk kinase complexes.

BACKGROUND OF THE INVENTION

Cellular Proliferation and Cancer

The disruption of external or internal regulation of cellular growth can lead to uncontrolled proliferation and in cancer, tumor formation. This loss of control can occur at many levels and, indeed, does occur at multiple levels in most tumors. Further, although tumor cells can no longer control their own proliferation, they still must use the same basic cellular machinery employed by normal cells to drive their growth and replication.

Cyclin Dependent Kinases and Cell Cycle Regulation

Progression of the normal cell cycle from the G1 to S phase, and from the G2 phase to M phase is dependent on cdks (Sherr, C. J.,Science274:1672-1677 (1996)). Like other kinases, cdks regulate molecular events in the cell by facilitating the transfer of the terminal phosphate of adenosine triphosphate (ATP) to a substrate protein. Isolated cdks require association with a second subunit, called cyclins (Desai et al.,Mol. Cell. Biol.,15:345-350 (1995)). Cyclins cause conformational changes at the cdk active site, allowing ATP access and interaction with the substrate protein. The balance between its rates of synthesis and degradation controls the level of each cyclin at any point in the cycle (Elledge, S. J., et al.,Biochim. Biophys. Acta,1377:M61-M70 (1998)). The influences of cyclin/cdk activity on the cell cycle and cellular transformation are summarized in Table 1.

Abnormal Cyclin/cdk Activity in Cancer

In a normal cell, interlocking pathways respond to the cell's external environment and internal checkpoints monitor conditions within the cell to control the activity of cyclin/cdk complexes. A reasonable hypothesis is that the disruption of normal control of cyclin/cdk activity may result in uncontrolled proliferation. This hypothesis appears to hold in a number of tumor types in which cyclins are expressed at elevated levels (Table 1). Mutations in the genes encoding negative regulators (proteins) of cyclin/cdk activity are also found in tumors (Larsen, C.-J.,Prog. Cell Cycle Res.,3:109-124 (1997)); (Kamb, A.,Trends in Genetics,11:136-140 (1995)). Members of the Cip family of cdk inhibitors form a ternary complex with the cyclin/cdk and require binding to cyclinA, cyclinE, or cyclinD (Hall, M., et al.,Oncogene,11:1581-1588 (1995)). In contrast, Ink family members form a binary complex with cdk4 or cdk6 and prevent binding to cyclinD (Parry, D.; et al.,EMBO J.,14:503-511 (1995)).

Tumors with elevated cyclin/cdk activity, whether from the over expression of cyclins or the loss of an endogenous cdk inhibitor, are prime targets for potential therapies based on small molecule cyclin/cdk inhibitors. In fact, several small molecule inhibitors of cyclin/cdks are reported (Meijer, L., et al., “Progress in Cell Cycle Research,”Plenum Press: New York,351-363 (1995)) and appear to bind at the ATP site of the kinase. Some information is known about small molecule inhibitors of other kinases, such as PKC (serine kinase) (Murray, K. J. et al., “Ann. Rep. Med. Chem.,” J. Bristol, Ed.,Academic Press, Inc.: New York,Chapter 26 (1994)) and tyrosine kinases (Fantl, W. J., et al.,Ann. Rev. Biochem.,62:453 (1993); Burke, T. R.,Drugs of the Future,17:119-1131 (1992); Dobrusin, E. M. et al., “Ann. Rep. Med. Chem,” J. Bristol, Ed.,Academic Press, Inc.: New York,Chapter 18 (1992); Spence, P.,Curr. Opin. Ther. Patents,3:3 (1993)). A number of known inhibitors were obtained from commercial sources or were synthesized by literature procedures.

Purine Compounds as Cyclin/cdk Inhibitors

The compounds of the present invention are shown to have far superior biological activities as cyclin/cdk complex inhibitors as well as inhibitors of cellular proliferation compared to those previously reported. In fact, the art (e.g., Fiorini, M. T. et al.,Tetrahedron Lett.,39:1827-1830 (1998)) teaches away from compounds of this invention, claiming lack of cellular proliferation inhibition.

SUMMARY OF THE INVENTION

The compounds of the present invention are 2,6,9-trisubstituted purine derivatives which are inhibitors of cyclin/cdk complexes. The compounds of the current invention also are potent inhibitors of human cellular proliferation. As such, the compounds of the present invention constitute pharmaceutical compositions with a pharmaceutically acceptable carrier. Such compounds are useful in treating conditions in a mammal mediated by elevated levels of cell proliferation compared to a healthy mammal by administering to such mammal an effective amount of the compound.

In one embodiment, the compounds of the present invention are represented by the chemical structure found in Formula I:
wherein:R1are the same or different and independently selected from the group consisting of:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3; andCH(CF3)2;X=N; orCH;V=NH;O;S; orCH2;R2=phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, and C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles selected from the group consisting of:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl; and4-isoquinolinyl; orsubstituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of Br, Cl, F, R1, and C(O)CH3;R3are the same or different and independently selected from the group consisting of:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; and(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R4=H;C1-C4-straight chain alkyl; orC3-C4-branched chain alkyl;R3and R4can be linked together by a carbon chain to form with intervening atoms a 5-8-membered saturated or unsaturated ring;n1=0-3;n=0-3;A=CH2;(CH2)2;(CH2)3;OCH2CH2; orCHCH3;Y=H;OR1;N(R1)2;N(R1)C(O)R3;N(R1)C(O)R5;N(R1)C(O)CH(R6)NH2;N(R1)SO2R3;N(R1)C(O)NHR3; orN(R1)C(O)OR6;R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; or(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;
or a pharmaceutically acceptable salt thereof;
with the proviso that the following conditions cannot all be true at the same time: R3=H, n=0, R4=H, and Y=OH.

Another aspect of the present invention is directed to a compound of the following formula:
wherein:R1are the same or different and independently selected from the group consisting of:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3; andCH(CF3)2;X=N; orCH;V=NH;O;S; orCH2;R2=phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, and C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles selected from the group consisting of:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl; and4-isoquinolinyl; orsubstituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of Br, Cl, F, R1, and C(O)CH3;n=0-3;A=CH2;(CH2)2;(CH2)3;OCH2CH2; orCHCH3;Y=H;OR1;N(R1)2;N(R1)C(O)R3;N(R1)C(O)R5;N(R1)C(O)CH(R6)NH2;N(R1)SO2R3;N(R1)C(O)NHR3; orN(R1)C(O)OR6;R3are the same or different and independently selected from the group consisting of:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; and(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alkyl; orC2-C4-alkenyl chain;
or a pharmaceutically acceptable salt thereof.

The present invention is also directed to a process for preparation of a purine derivative compound of the formula:
wherein:R1are the same or different and independently selected from the group consisting of:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3; andCH(CF3)2;X=N; orCH;V=NH;O;S; orCH2;R2=phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, and C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles selected from the group consisting of:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl; and4-isoquinolinyl; orsubstituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of Br, Cl, F, R1, and C(O)CH3;R3are the same or different and independently selected from the group consisting of:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; and(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R4=H;C1-C4-straight chain alkyl; orC3-C4-branched chain alkyl;R3and R4can be linked together by a carbon chain to form with intervening atoms a 5-8-membered saturated or unsaturated ring;n1=0-3;n=0-3;A=CH2;(CH2)2;(CH2)3;OCH2CH2; orCHCH3;Y=H;OR1;N(R1)2;N(R1)C(O)R3;N(R1)C(O)R5;N(R1)C(O)CH(R6)NH2;N(R1)SO2R3;N(R1)C(O)NHR3; orN(R1)C(O)OR6;R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; or(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;
or a pharmaceutically acceptable salt thereof;
with the proviso that the following conditions cannot all be true at the same time: R3=H, n=0, R4=H, and Y=OH, said process comprising:reacting a first intermediate compound of the formula:
where Z=Br or I;
with a compound of the formula: (R2—B(OH)2) or (R2—Sn(n-Bu)3or R2—SnMe3), or mixtures thereof, under conditions effective to form the purine derivative compound.

Another aspect of the present invention is directed to a process for preparation of a purine derivative compound of the formula:
wherein:R1are the same or different and independently selected from the group consisting of:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3; andCH(CF3)2;X=N; orCH;V=NH;O;S; orCH2;R2=phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, and C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles selected from the group consisting of:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl; and4-isoquinolinyl; orsubstituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of Br, Cl, F, R1, and C(O)CH3;R3are the same or different and independently selected from the group consisting of:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; and(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R4=H;C1-C4-straight chain alkyl; orC3-C4-branched chain alkyl;R3and R4can be linked together by a carbon chain to form with intervening atoms a 5-8-membered saturated or unsaturated ring;n1=0-3;n=0-3;A=CH2;(CH2)2;(CH2)3;OCH2CH2; orCHCH3;Y=H;OR1;N(R1)2;N(R1)C(O)R3;N(R1)C(O)R5;N(R1)C(O)CH(R6)NH2;N(R1)SO2R3;N(R1)C(O)NHR3; orN(R1)C(O)OR6;R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alky;C2-C4-alkenyl chain;(CH2)nPh; or(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;
or a pharmaceutically acceptable salt thereof;
with the proviso that the following conditions cannot all be true at the same time: R3=H, n=0, R4=H, and Y=OH, said process comprising:reacting a first intermediate compound of the formula:
under reductive or hydrogenation conditions effective to form the purine derivative compound.

Another aspect of the present invention is directed to a process for preparation of a purine derivative compound of the formula:
wherein:R1are the same or different and independently selected from the group consisting of:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3; andCH(CF3)2;X=N; orCH;V=NH;O;S; orCH2;R2=phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, and C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles selected from the group consisting of:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl; and4-isoquinolinyl; orsubstituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of Br, Cl, F, R1, and C(O)CH3;R3are the same or different and independently selected from the group consisting of:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; and(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R4=H;C1-C4-straight chain alkyl; orC3-C4-branched chain alkyl;R3and R4can be linked together by a carbon chain to form with intervening atoms a 5-8-membered saturated or unsaturated ring;n1=0-3;n=0-3;A=CH2;(CH2)2;(CH2)3;OCH2CH2; orCHCH3;Y=H;OR1;N(R1)2;N(R1)C(O)R3;N(R1)C(O)R5;N(R1)C(O)CH(R6)NH2;N(R1)SO2R3;N(R1)C(O)NHR3; orN(R1)C(O)OR6;R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; or(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;
or a pharmaceutically acceptable salt thereof;
with the proviso that the following conditions cannot all be true at the same time: R3=H, n=0, R4=H, and Y=OH, said process comprising:reacting a first intermediate compound of the formula:
with a compound of the formula:
where V1=NH2;OH; orSH;
under conditions effective to form the purine derivative compound.

Another aspect of the present invention is directed to a process for preparation of a purine derivative compound of the formula:
wherein:R1are the same or different and independently selected from the group consisting of:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3; andCH(CF3)2;X=N; orCH;V=NH;O;S; orCH2;R2=phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, and C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles selected from the group consisting of:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl; and4-isoquinolinyl; orsubstituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of Br, Cl, F, R1, and C(O)CH3;R3are the same or different and independently selected from the group consisting of:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; and(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R4=H;C1-C4-straight chain alkyl; orC3-C4-branched chain alkyl;R3and R4can be linked together by a carbon chain to form with intervening atoms a 5-8-membered saturated or unsaturated ring;n1=0-3;n=0-3;A=CH2;(CH2)2;(CH2)3;OCH2CH2; orCHCH3;Y=NR1C(O)R3NR1C(O)R5NR1SO2R3NR1C(O)NHR3NR1C(O)OR6R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; or(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;
or a pharmaceutically acceptable salt thereof, said process comprising:reacting a first intermediate compound having the same formula as the purine derivative compound except that Y═NHR1, with R3COCl or R5COCl or R3SO2Cl or R3NCO or R6OC(O)Cl under conditions effective to form the purine derivative compound.

Yet another aspect of the present invention is directed to a process for preparation of a purine derivative compound of the formula:
wherein:R1are the same or different and independently selected from the group consisting of:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3; andCH(CF3)2;X=N; orCH;V=NH;O;S; orCH2;R2=phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, and C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles selected from the group consisting of:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl; and4-isoquinolinyl; orsubstituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of Br, Cl, F, R1, and C(O)CH3;R3are the same or different and independently selected from the group consisting of:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; and(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R4=H;C1-C4-straight chain alkyl; orC3-C4-branched chain alkyl;R3and R4can be linked together by a carbon chain to form with intervening atoms a 5-8-membered saturated or unsaturated ring;n1=0-3;n=0-3;A=CH2;(CH2)2;(CH2)3;OCH2CH2; orCHCH3;Y=NHC(O)CH(R6)NH2R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; or(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;
or a pharmaceutically acceptable salt thereof; said process comprising:reacting a first intermediate compound having the same formula as the purine derivative compound except that Y is NH2, with PNHCH(R6)CO2H under conditions effective to form the purine derivative compound after a suitable deprotection strategy,
whereinP=C(O)OtBu;C(O)OCH2Ph;Fmoc;Benzyl; orAlloc.

The compounds of the present invention, as described in Formula I, show significantly improved growth inhibition of human transformed cell lines and/or cyclin/cdk inhibition relative to compounds of the prior art. These compounds have been demonstrated to be potent growth inhibitors in dozens of human transformed cell lines. Olomoucine, a structurally related purine derivative, is a poor human transformed cell growth inhibition agent with GI50values in the 20,000-100,000 nM range over 60-transformed cell lines. By contrast, the compounds of the present invention demonstrate GI50values over 60-transformed cell lines in the <10-25,000 nM range, preferably in the <10-100 nM range over 60-transformed cell lines, and, most preferably, <10 nM across 60-human transformed cell lines. This finding is unexpected from the prior art, which specifically teaches that compounds of the present invention would not be potent human transformed cell line growth inhibitors.

The R2group in Formula I imparts unexpected and significant improvement in growth inhibition in human transformed cell lines, while substitution of various groups at R3and R4found in Formula I impart important features that contribute to cyclin/cdk inhibition and growth inhibition of human transformed cell lines. Specifically, the combination of the R2group and the substitutions within R3and R4result in compounds with superior biological activity. Compounds which are cyclin/cdk inhibitors and/or human transformed cell line growth inhibitors have utility in treating human proliferative cellular disorders.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of the present invention are represented by the chemical structure found in Formula I.
wherein:R1are the same or different and independently selected from the group consisting of:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3; andCH(CF3)2;X=N; orCH;V=NH;O;S; orCH2;R2=phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, and C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles selected from the group consisting of:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl; and4-isoquinolinyl; orsubstituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of Br, Cl, F, R1, and C(O)CH3;R3are the same or different and independently selected from the group consisting of:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; and(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R4=H;C1-C4-straight chain alkyl; orC3-C4-branched chain alkyl;R3and R4can be linked together by a carbon chain to form with intervening atoms a 5-8-membered saturated or unsaturated ring;n1=0-3;n=0-3;A=CH2;(CH2)2;(CH2)3;OCH2CH2; orCHCH3;Y=H;OR1;N(R1)2;N(R1)C(O)R3;N(R1)C(O)R5;N(R1)C(O)CH(R6)NH2;N(R1)SO2R3;N(R1)C(O)NHR3; orN(R1)C(O)OR6;R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; or(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;
or a pharmaceutically acceptable salt thereof;
with the proviso that the following conditions cannot all be true at the same time: R3=H, n=0, R4=H, and Y=OH.

More preferably, the compounds of the current invention are represented by the chemical structure found in Formula III.
wherein:R1are the same or different and independently selected from the group consisting of:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3; andCH(CF3)2;X=N; orCH;V=NH;O;S; orCH2;R2=phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, and C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles selected from the group consisting of:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl; and4-isoquinolinyl; orsubstituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from the group consisting of Br, Cl, F, R1, and C(O)CH3;n=0-3;A=CH2;(CH2)2;(CH2)3;OCH2CH2; andCHCH3;Y=H;OR1;N(R1)2;N(R1)C(O)R3;N(R1)C(O)R5;N(R1)C(O)CH(R6)NH2;N(R1)SO2R3;N(R1)C(O)NHR3; orN(R1)C(O)OR6;R3are the same or different and independently selected from the group consisting of:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh; and(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alkyl; orC2-C4-alkenyl chain;
or a pharmaceutically acceptable salt thereof.

In another embodiment, the present invention is directed to a method for inhibiting cellular proliferation in mammals comprising administering a therapeutically effective amount of the compound of the present invention to the mammal.

The compounds of the present invention can be administered orally, parenterally, for example, subcutaneously, intravenously, intramuscularly, intraperitoneally, by intranasal instillation, or by application to mucous membranes, such as, that of the nose, throat, and bronchial tubes. They may be administered alone or with suitable pharmaceutical carriers, and can be in solid or liquid form such as, tablets, capsules, powders, solutions, suspensions, or emulsions.

Based on the results obtained in the standard pharmacological test procedures described below, the compounds of the present invention are useful as antineoplastic agents. More particularly, the compounds of the present invention are useful for inhibiting the growth of neoplastic cells, causing cell death of neoplastic cells, and eradicating neoplastic cells. The compounds of the present invention are, therefore, useful for treating solid tumors, including sarcomas and carcinomas, such as astrocytomas, prostate cancer, breast cancer, small cell lung cancer, and ovarian cancer, leukemias, lymphomas, adult T-cell leukemia/lymphoma, and other neoplastic disease states.

In addition to the utilities described above, many of the compounds of the present invention are useful in the preparation of other compounds.

The active compounds of the present invention may be orally administered, for example, with an inert diluent, or with an assimilable edible carrier, or they may be enclosed in hard or soft shell capsules, or they may be compressed into tablets, or they may be incorporated directly with the food of the diet. For oral therapeutic administration, these active compounds may be incorporated with excipients and used in the form of tablets, capsules, elixirs, suspensions, syrups, and the like. Such compositions and preparations should contain at least 0.1% of active compound. The percentage of the compound in these compositions may, of course, be varied and may conveniently be between about 2% to about 60% of the weight of the unit. The amount of active compound in such therapeutically useful compositions is such that a suitable dosage will be obtained. Preferred compositions according to the present invention are prepared so that an oral dosage unit contains between about 1 and 250 mg of active compound.

Various other materials may be present as coatings or to modify the physical form of the dosage unit. For instance, tablets may be coated with shellac, sugar, or both. A syrup may contain, in addition to active ingredient, sucrose as a sweetening agent, methyl and propylparabens as preservatives, a dye, and flavoring such as cherry or orange flavor.

These active compounds may also be administered parenterally. Solutions or suspensions of these active compounds can be prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof in oils. Illustrative oils are those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, or mineral oil. In general, water, saline, aqueous dextrose and related sugar solution, and glycols such as, propylene glycol or polyethylene glycol, are preferred liquid carriers, particularly for injectable solutions. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

The compounds of the present invention may also be administered directly to the airways in the form of an aerosol. For use as aerosols, the compounds of the present invention in solution or suspension may be packaged in a pressurized aerosol container together with suitable propellants, for example, hydrocarbon propellants like propane, butane, or isobutane with conventional adjuvants. The materials of the present invention also may be administered in a non-pressurized form such as in a nebulizer or atomizer.

General Synthetic Schemes

The compounds of the present invention can be prepared by conventional methods of organic synthesis practiced by those skilled in the art. The general reaction sequences outlined below are general methods useful for preparing the compounds of the present invention and are not meant to be limiting in scope or utility.

Reaction of 2,6-dichloropurine (Formula IV) with various amines of Formula V, many of which are commercially available or prepared by literature methods or modifications of literature methods, in the presence of a polar solvent, such as ethanol, provides purines of Formula VI (General Flowsheet I, infra). Reaction of purines of Formula VI with alkyl halides (R1—Z) in the presence of a base such as potassium carbonate provides N1-alkylated purines of Formula VII. Chloride displacement of N1-alkylated purines of Formula VII with amines, thiols or alcohols of structure Formula VIII, either in neat solution or in an inert solvent such as ethanol or butanol, with or without a base such as sodium hydride as appropriate, at an appropriate temperature provides purines of Formula IX (V=NH, O, S). Transition metal-mediated cross-coupling reaction of purines of Formula IX with boronic acid (R2—B(OH)2) or tin reagents (R2—Sn(n-Bu)3or R2—SnMe3) provides purines of Formula X (V=NH, O, S). If in Formula X (Y=NH2), then subsequent reaction of Formula X (Y=NH2) with acid chloride (R3COCl), or sulfonyl chloride (R3SO2Cl), or isocyanate (R3NCO), or chloroformate (ClC(O)OR6) reagents provides purines of Formula XI wherein Y=NHC(O)R3, NHSO2R3, or NHC(O)NHR3, or NHC(O)OR6, respectively. On the other hand, if in Formula X, Y already is OR1or NHC(O)R3or NHSO2R3or NHC(O)NHR3or NHC(O)OR6, as a result of what Y started out as in Formula VIII, then this last step is unnecessary.

Reaction of purines of Formula VII, with alkenyl tin reagents of Formula XII, which are prepared by conventional methods described in the literature, in the presence of a transition metal catalyst, such as Pd(0), provides purines of Formula XIII (General Flowsheet II, infra). Subsequent reaction of purines of Formula XIII with boronic acid (R2—B(OH)2) or tin reagents (R2—Sn(n-Bu)3or R2—SnMe3) in the presence of a transition metal catalyst, such as Pd(0), provides purines of Formula XV. Alternatively, by switching the order of reactions dependent on the precise reactivity of the purine of Formula VII, reaction of purines of Formula VII with boronic acid (R2—B(OH)2) or tin reagents (R2—Sn(n-Bu)3or R2—SnMe3) in the presence of a transition metal catalyst, such as Pd(0), provides purines of Formula XIV. Subsequent reaction of purines of Formula XIV, with alkenyl tin reagents of Formula XII, which are prepared by conventional methods described in the literature, in the presence of a transition metal catalyst, such as Pd(0), provides purines of Formula XV. Finally reduction of the olefin within Formula XV provides purines of Formula X (V=CH2).

Definitions of the groups include:

Z=Br;I;V1=NH2;OH;SH;R1are the same or different and independently selected from:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3;CH(CF3)2;X=N;CH;V=NH;O;S;CH2;R2can be in any position on the ring and selected from:phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles including:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl;4-isoquinolinyl;substituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from Br, Cl, F, R1, C(O)CH3;R3are the same or different and independently selected from:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh;(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R4=H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;R3and R4can be linked together by a carbon chain to form a 5-8-membered saturated or unsaturated ring;n1=0-3;n=0-3;A=(CH2);(CH2)2;(CH2)3;(OCH2CH2);(CHCH3);Y=H;OR1;N(R1)2;N(R1)C(O)R3;N(R1)C(O)R5;N(R1)C(O)CH(R6)NH2;N(R1)SO2R3;N(R1)C(O)NHR3;N(R1)C(O)OR6;R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh;(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2.

Additional, general non-limiting syntheses of compounds of the present invention of Formula X and Formula XI are shown below in General Flowsheet III.
Reaction of various amines of Formula V, many of which are commercially available or prepared by literature methods or modifications of literature methods, with boronic acid (R2—B(OH)2) or tin reagents (R2—Sn(n-Bu)3) or (R2—SnMe3) in the presence of a transition metal catalyst, such as Pd(0), provides biaryl amines of Formula XVII. Reaction of 2,6-dichloropurine (Formula IV) with various amines of Formula XVII, in the presence of a polar solvent, such as ethanol, provides purines of Formula XVIII. Reaction of purines of Formula XVIII with alkyl halides (R1—Z) in the presence of a base such as potassium carbonate provides N1-alkylated purines of Formula XIV. Chloride displacement of N1-alkylated purines of Formula XIV with amines, thiols or alcohols of Formula VIII, either in neat solution or in an inert solvent such as ethanol or butanol, with or without a base such as sodium hydride as appropriate, at an appropriate temperature provides purines of Formula X (V=NH, O, S). If in Formula X (Y=NH2), then subsequent reaction of Formula X (Y=NH2) with acid chloride (R3COCl), or sulfonyl chloride (R3SO2Cl), or isocyanate (R3NCO), or chloroformate (ClC(O)OR6) reagents provides purines of Formula XI wherein Y=NHC(O)R3, NHSO2R3, or NHC(O)NHR3, or NHC(O)OR6, respectively. On the other hand, if in Formula X, Y already is OR1or NHC(O)R3or NHSO2R3or NHC(O)NHR3or NHC(O)OR6, as a result of what Y started out as in Formula VIII, then this last step is unnecessary.
Definitions of the groups include:Z=Br;I;V1=NH2;OH;SH;R1are the same or different and independently selected from:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3;CH(CF3)2;X=N;CH;V=NH;O;S;CH2;R2can be in any position on the ring and selected from:phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles including:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl;4-isoquinolinyl;substituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from Br, Cl, F, R1, C(O)CH3;R3are the same or different and independently selected from:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh;(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R4=H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;R3and R4can be linked together by a carbon chain to form a 5-8-membered saturated or unsaturated ring;n1=0-3;n=0-3;A=(CH2);(CH2)2;(CH2)3;(OCH2CH2);(CHCH3);Y=H;OR1;N(R1)2;N(R1)C(O)R3;N(R1)C(O)R5;N(R1)C(O)CH(R6)NH2;N(R1)SO2R3;N(R1)C(O)NHR3;N(R1)C(O)OR6;R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh;(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2.
Additional, general non-limiting syntheses of compounds of the present invention of Formula XVI, Formula XVII and Formula XVIII are shown below in General Flowsheet IV.

If in Formula X (Y=NH2), then subsequent reaction of Formula X (Y=NH2) with alkyl halide (R8CH2Z), an appropriate base, and a solvent; or reaction of Formula X (Y=NH2) with aldehyde (R8CHO) in the presence of a solvent and a suitable reducing agent provides purines of Formula XVI wherein Y=NHR1, or N(R1)2. On the other hand, if in Formula X, Y already is NHR1, or N(R1)2, as a result of what Y started out as in Formula X, then this last step is unnecessary. If in Formula XVI (Y=NHR1), then subsequent reaction of Formula XVI (Y=NHR1) with acid chloride (R3COCl or R5COCl), or sulfonyl chloride (R3SO2Cl), or isocyanate (R3NCO), or chloroformate (ClC(O)OR6) reagents provides purines of Formula XX wherein Y=NR1C(O)R3, or NR1C(O)R5or NR1SO2R3, or NR1C(O)NHR3, or NR1C(O)OR6, respectively. On the other hand, if in Formula XVI, Y already is NR1C(O)R3, or NR1C(O)R5, or NR1SO2R3, or NR1C(O)NHR3, or NR1C(O)OR6, as a result of what Y started out as in Formula XVI, then this last step is unnecessary.

If in Formula X (Y=NH2), then subsequent reaction of Formula X (Y=NH2) with acid (PNHCH(R6)CO2H), in a suitable solvent in the presence of an appropriate coupling agent provides a purine derivative; which upon suitable deprotection provides purines of Formula XIX wherein Y=NHC(O)CH(R6)NH2. On the other hand, if in Formula X, Y already is NHC(O)CH(R1)NH2, as a result of what Y started out as in Formula X, then this last step is unnecessary.

Definitions of the groups include:

Z=Br;I;P=C(O)OtBu;C(O)OCH2Ph;Fmoc;Benzyl;Alloc;R1are the same or different and independently selected from:H;C1-C6-straight chain alkyl;C2-C6-straight alkenyl chain;C3-C6-branched alkyl chain;C3-C6-branched alkenyl chain;C3-C7-cycloalkyl;CH2—(C3-C7-cycloalkyl);CH2CF3;CH2CH2CF3;CH(CF3)2;X=N;CH;V=NH;O;S;CH2;R2can be in any position on the ring and selected from:phenyl;substituted phenyl, wherein the substituents (1-2 in number) are in any position and are independently selected from R1, OR1, SR1, S(O)R1, S(O2)R1, NHR1, NO2, OC(O)CH3, NHC(O)CH3, F, Cl, Br, CF3, C(O)R1, C(O)NHR1, phenyl, C(O)NHCHR1CH2OH;1-naphthyl;2-naphthyl;heterocycles including:2-pyridyl;3-pyridyl;4-pyridyl;2-pyrimidyl;4-pyrimidyl;5-pyrimidyl;thiophene-2-yl;thiophene-3-yl;2-furanyl;3-furanyl;oxazol-2-yl;oxazol-4-yl;oxazol-5-yl;thiazol-2-yl;thiazol-4-yl;thiazol-5-yl;imidazol-2-yl;imidazol-4-yl;pyrazol-3-yl;pyrazol-4-yl;isoxazol-3-yl;isoxazol-4-yl;isoxazol-5-yl;isothiazol-3-yl;isothiazol-4-yl;isothiazol-5-yl;1,3,4-thiadiazol-2-yl;benzo[b]furan-2-yl;benzo[b]thiophene-2-yl;2-pyrrolyl;3-pyrrolyl;1,3,5-triazin-2-yl;pyrazin-2-yl;pyridazin-3-yl;pyridazin-4-yl;2-quinolinyl;3-quinolinyl;4-quinolinyl;1-isoquinolinyl;3-isoquinolinyl;4-isoquinolinyl;substituted heterocycle, wherein the substituents (1-2 in number) are in any position and are independently selected from Br, Cl, F, R1, C(O)CH3;R3are the same or different and independently selected from:H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh;(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R4=H;C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;R3and R4can be linked together by a carbon chain to form a 5-8-membered saturated or unsaturated ring;n1=0-3;n=0-3;A=(CH2);(CH2)2;(CH2)3;(OCH2CH2);(CHCH3);Y=H;OR1;N(R1)2;N(R1)C(O)R3;N(R1)C(O)R5;N(R1)C(O)CH(R6)NH2;N(R1)SO2R3;N(R1)C(O)NHR3;N(R1)C(O)OR6;R5=C3-C7-cycloalkyl;R6=C1-C4-straight chain alkyl;C3-C4-branched chain alkyl;C2-C4-alkenyl chain;(CH2)nPh;(CH2)n-substituted phenyl, wherein the phenyl substituents are as defined above in R2;R8=C1-C5-straight chain alkyl;C2-C5-straight alkenyl chain;C3-C5-branched alkyl chain;C3-C5-branched alkenyl chain;C3-C7-cycloalkyl;CF3;CH2CF3.
The synthesis of compound 5 is shown below in Scheme I.
The synthesis of compound 11 is shown below in Scheme II.
The syntheses of compounds 12, 13 and 14 are shown below in Scheme III.
The synthesis of compound 17 is shown below in Scheme IV.
The synthesis of compound 17 is shown below in Scheme V.
The synthesis of compound 25 is shown below in Scheme VI.
An alternative synthesis of compound 25 is shown below in Scheme VII.
The synthesis of compound 32 is shown below in Scheme VIII.
The syntheses of compounds 33 and 34 are shown below in Scheme IX.
The syntheses of compounds 36, 38, and 40 are shown below in Scheme X.
The synthesis of compound 43 is shown below in Scheme XI.
The synthesis of compound 46 is shown below in Scheme XII.
The syntheses of compound 48 and 50 are shown below in Scheme XIII.
The synthesis of compound 53 is shown below in Scheme XIV.
The synthesis of compound 54 is shown below in Scheme XV.
The synthesis of compound 56 is shown below in Scheme XVI.
The synthesis of compound 58 is shown below in Scheme XVII.
The synthesis of compound 60 is shown below in Scheme XVIII.
The syntheses of compounds 61, and 62 are shown below in Scheme XIX.
The syntheses of compounds 64, and 65 are shown below in Scheme XX.
The syntheses of compounds 66, and 67 are shown below in Scheme XXI.
The synthesis of compound 73 is shown below in Scheme XXII.
The syntheses of compounds 74, 75, and 76 are shown below in Scheme XXIII.
The synthesis of compound 77 is shown below in Scheme XXIV.
The synthesis of compound 78 is shown below in Scheme XXV.
An alternative synthesis of compound 78, and the synthesis of compound 79 are shown below in Scheme XXVI.
The synthesis of compound 80 is shown below in Scheme XXVII.
The synthesis of compounds 86, and 87 are shown below in Scheme XXVIII.
The synthesis of compound 88 is shown below in Scheme XXIX.
The syntheses of compounds 93, and 94 are shown below in Scheme XXX.
The syntheses of compounds 95, and 96 are shown below in Scheme XXXI.
The synthesis of compound 97 is shown below in Scheme XXXII.
The syntheses of compounds 98, and 99 are shown below in Scheme XXXIII.
The synthesis of compound 100 is shown below in Scheme XXXIV.
The syntheses of compounds 101, and 102 are shown below in Scheme XXXV.
The syntheses of compounds 103, and 104 are shown below in Scheme XXXVI.
The syntheses of compounds 106, 107, and 108 are shown below in Scheme XXXVII.
The syntheses of compounds 109, and 110 are shown below in Scheme XXXVIII.
The syntheses of compounds 111, and 112 are shown below in Scheme XXXIX.
The synthesis of compound 113 is shown below in Scheme XL.
The syntheses of compounds 114, 115, 116, and 117 are shown below in Scheme XLI.
The synthesis of compound 118 is shown below in Scheme XLII.
The syntheses of compounds 123 and 124 are shown below in Scheme XLIII.
The syntheses of compounds 131 and 132 are shown below in Scheme XLIV.
The syntheses of compounds 134 and 135 are shown below in Scheme XLV.
The synthesis of compound 137 is shown below in Scheme XLVI.
The syntheses of compound 139 and 140 are shown below in Scheme XLVII.
The synthesis of compound 142 is shown below in Scheme XLVIII.
The synthesis of compound 144 is shown below in Scheme XLIX.
The synthesis of compound 146 is shown bellow in Scheme L.
The synthesis of compound 148 is shown below in Scheme LI.
The syntheses of compounds 149-152 are shown below in Scheme LII.
The syntheses of compounds 153-156 are shown below in Scheme LIII.
The syntheses of compounds 157-159 are shown below in Scheme LIV.
The syntheses of compounds 160-163 are shown below in Scheme LV.
The syntheses of compounds 164-166 are shown below in Scheme LVI.
The syntheses of compounds 167-168 are shown below in Scheme LVII.
The syntheses of compounds 169-171 are shown below in Scheme LVIII.
The syntheses of compounds 172-173 are shown below in Scheme LIX.
The syntheses of compounds 174-176 are shown below in Scheme LX.
The syntheses of compounds 177-178 are shown below in Scheme LXI.
The syntheses of compounds 179-180 are shown below in Scheme LXII.
The syntheses of compounds 181-182 are shown below in Scheme LXIII.
The syntheses of compounds 187-188 are shown below in Scheme LXIV.
The syntheses of compounds 193 and 194 are shown below in Scheme LXV.
The syntheses of compounds 199-200 are shown below in Scheme LXVI.
The syntheses of compounds 205-206 are shown below in Scheme LXVII.
The syntheses of compounds 207-210 are shown below in Scheme LXVIII.
The syntheses of compounds 211-212 are shown below in Scheme LXIX.
The syntheses of compounds 213-215 are shown below in Scheme LXX.
The syntheses of compounds 216-217 are shown below in Scheme LXXI.
The syntheses of compounds 218-219 are shown below in Scheme LXXII.
The synthesis of compounds 221 is shown below in Scheme LXXIII.
The synthesis of compound 222 is shown below in Scheme LXXIV.
The synthesis of compound 223 is shown below in Scheme LXXV.
The synthesis of compound 224 is shown below in Scheme LXXVI.
The synthesis of compound 229 is shown below in Scheme LXXVII.
The syntheses of compounds 230-233 are shown below in Scheme LXXVIII.
The syntheses of compounds 239-241 are shown below in Scheme LXXIX.
The syntheses of compounds 242-243 are shown below in Scheme LXXX.
The syntheses of compounds 248-250 are shown below in Scheme LXXXI.
The syntheses of compounds 123 and 124 are shown below in Scheme LXXXII.
The syntheses of compounds 258-260 are shown below in Scheme LXXXIII.
The syntheses of compounds 261-263 are shown below in Scheme LXXXIV.
The syntheses of compounds 264-265 are shown below in Scheme LXXXV.
The synthesis of compound 266 is shown below in Scheme LXXXVI.

EXAMPLES

Proton NMR spectra were obtained on a Bruker AC 300 spectrometer at 300 MHz or a Bruker 500 MHz spectrometer and were referenced to tetramethylsilane as an internal standard. The IR spectrometer used was a single beam Perkin-Elmer Spectrum 1000 FT-IR. All IR spectra obtained were prepared in a pressed disc of KBr. All IR spectra obtained were acquired with a total of 4 accumulations at a resolution of 4.00 cm−1. Melting points were obtained on a Mel-Temp II apparatus and are uncorrected. Mass spectra were obtained on either a Shimadzu QP-5000 or a PE Sciex API 150 Mass Spectrometer.

Preparation of Compound 2

Preparation of Compound 3

Preparation of Compound 4

Preparation of Compound 5

Preparation of Compound 7

To 4-iodobenzoic acid (52.2 g, 0.21 mol) was added CH2Cl2(500 mL) and DMF (2 drops) at room temperature. Oxalyl chloride (32 g, 0.25 mol) was added dropwise in 0.5 h and stirred for 2 d. The volatiles were removed in vacuo to a volume of 150 mL to give the acid chloride and CH2Cl2. To a mixture of ice (500 mL) and NH4OH (29%; 100 mL) was added the CH2Cl2solution during 15 min. The resulting solids were collected, washed with CH2Cl2, and dried in vacuo. The solids were slurried in H2O for 1 h. The solids were collected by filtration, washed in water and acetone, and dried in vacuo to give 7 (48 g; 92%): mp 213-216° C.

Preparation of Compound 8

Preparation of Compound 9

Preparation of Compound 10

Preparation of Compound 11

Preparation of Compound 12

Preparation of Compound 13

Preparation of Compound 14

Preparation of Compound 16

A solution of 15 (2.5 g, 15.8 mmol) and ether was cooled to −78° C. In a separate flask, n-BuLi (15.8 mmol) was also cooled to −78° C. The solution of 15 was added to the n-BuLi solution via cannula to give a dark red solution. The reaction mixture was stirred for 5 min prior to the rapid addition of (n-Bu)3SnCl (6.2 g, 19 mmol). The resulting bright yellow solution was stirred at −78° C. for 2 h, allowed to warm to room temperature, and stirred for another 10 min. The solution was then diluted with H2O (80 mL) and extracted with ethyl acetate (3×50 mL). The organic extracts were combined, washed with brine, dried over Na2SO4, filtered, and concentrated in vacuo to yield the crude product as a yellow oil. Purification by column chromatography gave the product 16 (4.89 g, 84%) as a pale yellow liquid:1H NMR (300 MHz, CDCl3) δ 8.72 (d, 1 H), 7.48-7.46 (m, 1 H), 7.40-7.38 (m, 1 H), 7.11-7.09 (m, 1 H), 1.61-1.50 (m, 6 H), 1.38-1.26 (m, 6 H), 1.14-1.09 (m, 6 H), 0.97-0.77 (t, 9 H).

Preparation of Compound 17

Preparation of Compound 19

To a solution of n-BuLi (2.5M hexane solution, 10.9 mL, 27.4 mmol) in ethyl ether 28 mL at −78° C. was added 2-bromopyridine (4.33 g, 27.4 mmol) in ethyl ether (15 mL). After stirring for 30 min, a solution of trimethylstannylchloride (6.0 g, 30 mmol) in THF (10 mL) was added. Stirring was continued at −78° C. for 2 h and the mixture was then warmed up to room temperature and filtered. The precipitate was washed with ether and the combined the ether filtrates were concentrated to give the crude product:1H NMR (500 Hz, CDCl3) δ 8.69-8.68 (d, 1 H), 7.47-7.07 (m, 3 H), 0.30 (s, 9 H).

Preparation of Compound 21

Preparation of Compound 22

To LiAlH4(8 mmol) in THF (25 mL) was added 21 (0.96 g, 5.3 mmol) in THF (15 mL) slowly while the flask was cooled with ice. The mixture was stirred at room temperature for 10-30 min then stirred at reflux for 4 h under nitrogen. The mixture was cooled in an ice bath and aqueous sodium hydroxide solution (0.5 mL, 10%) was added. The mixture was stirred until the residue became white and the solid was filtered and washed with methylene chloride (4×5 mL). The methylene chloride solution was dried with anhydrous sodium sulfate, concentrated, and the crude product was chromatographed on silica gel to give the product as a yellow liquid. A small amount of ethanol was added and the pure amine 22 was obtained as a white solid (74%) after filtration: mp 114-117° C.;1H NMR (500 Hz, CDCl3) δ 8.66 (d, J=4.4 Hz, 1 H), 7.94 (d, J=8.1 Hz, 2 H), 7.70 (m, 2 H), 7.39 (d, J=8.0 Hz), 7.19 (m, 1 H), 3.90 (s, 2 H), 1.98 (s, 2 H).

Preparation of Compound 23

Preparation of Compound 24

Preparation of Compound 17

Preparation of Compound 25

Preparation of Compound 27

A mixture of diethyl(3-pyridyl)borane (26, 540 mg, 3.67 mmol), 4-bromobenzonitrile (803 mg, 4.41 mmol) and Pd(PPh3)4(144 mg, 0.13 mmol) in toluene (9 mL), ethanol (1.3 mL) and 2M aqueous sodium carbonate solution (4.1 mL, 8.2 mmol) was heated at 90-100° C. under nitrogen for 27 h. The mixture was cooled to room temperature and water (10 mL) was added. The organic layer was separated and the aqueous layer was extracted with ethyl acetate (2×20 mL). The combined organic layers were washed with brine (2×15 mL) and dried over anhydrous sodium sulfate. Flash chromatography of the crude product on silica gave the product as a white solid (80%): mp 95-96° C.

An Alternative Preparation of 27 is Described Below

Preparation of Compound 28

To LiAlH4(8 mmol) in THF (25 mL) was added 27 (0.96 g, 5.3 mmol) in THF (25 mL) slowly while the flask was cooled with ice. The mixture was stirred at room temperature for 10-30 min then stirred at reflux for 4 h under nitrogen. The mixture was cooled in an ice bath and aqueous sodium hydroxide solution (0.5 mL, 10%) was added. The mixture was stirred until the residue became white and the solid was filtered and washed with methylene chloride (4×5 mL). The methylene chloride solution was dried with anhydrous sodium sulfate, concentrated, and the crude product was chromatographed on silica gel to give the product as a yellow liquid. A small amount of ethanol was added and the pure amine 28 was obtained as a white solid (46%) after filtration: mp 94-96° C.;1H NMR (500 Hz, CDCl3) δ 8.74 (d, J=2.4 Hz, 1 H), 8.48 (dd, J1=1.5 Hz, J2=4.7 Hz, 1 H), 7.77 (m, 1 H), 7.45 (d, J=8.10 Hz, 2 H), 7.33 (d, J=8.0 Hz, 2 H), 7.25 (m, 1 H), 3.83 (s, 2 H), 2.25 (s, 2 H).

Preparation of Compound 29

Preparation of Compound 30

Preparation of Compound 32

Preparation of Compound 33

Preparation of Compound 34

Preparation of Compound 36

Preparation of Compound 38

Preparation of Compound 40

Preparation of Compound 43

Preparation of Compound 45

To a solution of s-BuLi (5 mL, 6.24 mmol) and TMEDA (1 mL) in anhydrous THF (35 mL) at −75° C. under argon was added dropwise a solution of N,N-diethylbenzamide (0.98 g, 5.57 mmol) in THF (5 mL). The mixture was stirred for 50 min and then treated with trimethylborate (2 mL, 17 mmol). The solution was allowed to warm to room temperature overnight. The colorless solution was cooled to 0° C. and acidified to pH=6 with 2N HCl. The THF was removed in vacuo and the residue was diluted with water. This was extracted with CH2Cl2(3×50 mL) and the combined organic layers were washed with brine, dried over Na2SO4, concentrated in vacuo, followed by removal of trace solvent on the vacuum pump to give 45 as an off-white foamy solid:1H NMR (300 MHz, CD3OD) δ 7.67-7.39 (m, 4 H), 3.88-3.69 (q, 4 H), 1.41-1.30 (t, 6 H).

Preparation of Compound 46

Preparation of Compound 48

Preparation of Compound 50

Preparation of Compound 52

Preparation of Compound 53

Preparation of Compound 54

To compound 3 (0.26 g, 0.67 mmol) was added trans-4-aminocyclohexanol hydrochloride (0.62 g, 4.11 mmol), Et3N (0.58 mL, 4.16 mmol), and ethanol (5 mL). The mixture was heated for 5 h at 135° C. in an oil bath. The temperature increased to 150° C. and heating was continued for a further 48 h. The solution was cooled and evaporated to give a yellow oil: CI MS m/z=459 [C21H27BrN6O+H]+.

Preparation of Compound 55

Preparation of Compound 56

Preparation of Compound 57

Preparation of Compound 58

Preparation of Compound 59

Preparation of Compound 60

Preparation of Compound 61

Preparation of Compound 62

Preparation of Compound 64

Preparation of Compound 65

Preparation of Compound 66

Preparation of Compound 67

Preparation of Compound 69

To a solution of 4-biphenylcarboxaldehyde (1.0 g, 5.49 mmol) in MeOH (20 mL) was added NaBH3CN (0.69 g, 11.0 mmol), and NH4OH (15 mL) and the mixture was stirred at room temperature overnight. To this added HCl and extracted with CHCl3. The resulting aqueous layer was brought to pH>7 with sodium bicarbonate and then extracted with CHCl3. The solution was dried with MgSO4, filtered, and evaporated to give 69 (200 mg) as a white solid: EI MS m/z=183 [C13H13N]+.

Preparation of Compound 69

To compound 70 (2.75 g, 13.9 mmol) was added anhydrous THF (60 mL), heated to reflux, and kept under nitrogen. 1M Borane-THF (69.7 mL) was added dropwise to 70 through an addition funnel resulting in a homogeneous solution. The solution was refluxed for 18 h. The reaction mixture was cooled in an ice water bath and quenched with H2O, 2N HCl (20 mL), followed by 3N NaOH (60 mL). The reaction mixture was extracted with EtOAc (3×). The organic extracts were washed with brine, and dried over sodium sulfate. The crude product was concentrated, dissolved in MeOH, and HCl gas was bubbled through the solution. The solution was filtered in vacuo to give 69 as a white solid:1H NMR (300 MHz, CD3OD) δ 7.71 (d, 2 H), 7.63 (d, 2 H), 7.52 (d, 2 H), 7.47-7.30 (m, 3 H), 4.13 (s, 2 H).

Preparation of Compound 71

To compound 1 (6.8 g, 36.0 mmol) and 69 (8.0 g, 36.5 mmol) was added H2O (60 mL) and Hünigs base (9.0 g, 70.0 mmol). The mixture was stirred and heated to reflux for 5 h during which time H2O (50 mL) was added as the reaction continued to thicken. The crude product was collected by filtration, washed with H2O (500 mL) and EtOH (2×30 mL), air dried, and dried in vacuo to give 71 (11.1 g, 92%): mp 267-269° C.

Preparation of Compound 72

Preparation of Compound 73

Preparation of Compound 74

Preparation of Compound 75

Preparation of Compound 76

Preparation of Compound 77

Preparation of Compound 78

Preparation of Compound 78

Preparation of Compound 79

Preparation of Compound 80

Preparation of Compound 82

Preparation of Compound 83

Preparation of Compound 84

Preparation of Compound 85

Preparation of Compound 86

Preparation of Compound 87

Preparation of Compound 88

Preparation of Compound 89

Preparation of Compound 90

Preparation of Compound 91

Preparation of Compound 92

Preparation of Compound 93

Preparation of Compound 94

Preparation of Compound 95

Preparation of Compound 96

Preparation of Compound 97

Preparation of Compound 98

Preparation of Compound 99

Preparation of Compound 100

Preparation of Compound 101

Preparation of Compound 102

Preparation of Compound 103

Preparation of Compound 104

Preparation of Compound 106

Preparation of Compound 107

Preparation of Compound 108

Preparation of Compound 109

Preparation of Compound 110

Preparation of Compound 111

Preparation of Compound 112

Preparation of Compound 113

Preparation of Compound 114

Preparation of Compound 115

Preparation of Compound 117

Preparation of Compound 116

Preparation of Compound 118

Preparation of Compound 119

Preparation of Compound 120

Preparation of Compound 121

Preparation of Compound 122

Preparation of Compound 123

Preparation of Compound 124

Alternative Preparation of Compound 71

To a solution of 4-phenylbenzoic acid (5.46 g, 27.6 mmol) in methylene chloride (66 mL) was added 2 drops of DMF and oxalyl chloride (2.80 mL, 30.3 mmol). The reaction mixture was stirred overnight and added to a stirred solution of ice and ammonium hydroxide. The resulting precipitate was filtered, washed with methylene chloride, and triturated with water. The product was collected by filtration and dried in vacuo to yield 4-phenylbenzamide (3.88 g, 71%).

Under a nitrogen atmosphere, 4-phenylbenzamide (2.01 g, 10.2 mmol) was dissolved in THF (50 mL) and heated to reflux. To the mixture was added dropwise 1 M borane in THF (80.0 mL, 80.0 mmol). After refluxing for 18 h, the reaction mixture was cooled to room temperature and treated with 1 N HCl (40 mL). The solution was made basic via addition of 3 N NaOH (60 mL) and extracted with ethyl acetate (3×370 mL). The extract was washed with brine and dried over sodium sulfate. Concentration yielded 4-phenylbenzyl amine as a white solid (1.73 g, 93%).

Preparation of Compound 126

Preparation of Compound 127

Preparation of Compound 128

Preparation of Compound 129

Preparation of Compound 130

Preparation of Compound 131

Preparation of Compound 132

Preparation of Compound 133

Preparation of Compound 134

Preparation of Compound 135

Preparation of Compound 136

To a mixture of 71 (2.00 g, 5.96 mmol) in dimethylsulfoxide (44 mL) was added potassium carbonate (6.56 g, 47.7 mmol) and iodoethane (2.00 mL, 24.4 mmol). After stirring overnight, the reaction mixture was poured into a stirred solution of water (300 mL). After 2 d, it was filtered. The filtrate was extracted with ethyl acetate (2×180 mL). The organic extracts were combined and washed with brine (150 mL). The organic layer was dried over magnesium sulfate. Concentration afforded 136 (1.90 g, 88%).

Preparation of Compound 137

Preparation of Compound 138

Compound 71 (2.02 g, 6.02 mmol), iodomethane (1.50 mL, 24.4 mmol), and potassium carbonate were dissolved in dimethylsulfoxide (44 mL) and stirred overnight. The reaction mixture was poured into 150 mL of stirring water. The organic and aqueous layers were separated. The organic layer was washed with brine (3×100 mL) and dried over magnesium sulfate. The solids were removed by filtration and the solution was concentrated in vacuo to afford 138 (1.93 g, 93%).

Preparation of Compound 139

Preparation of Compound 140

Preparation of Compound 144

Compound 71 (2.03 g, 5.96 mmol), 1-iodopropane (2.25 mL, 24.4 mmol), and potassium carbonate (6.61 g, 47.7 mmol) were dissolved in dimethylsulfoxide (44 mL) and allowed to stir overnight. The reaction mixture was added to 300 mL of stirring water and stirred for 2 d. The resulting precipitate was collected by filtration and dried in vacuo to afford 141 (2.07 g, 92%).

Preparation of Compound 142

Preparation of Compound 143

Compound 71 (2.01 g, 5.98 mmol), iodocyclopentane (2.80 mL, 24.2 mmol), and potassium carbonate (6.75 g, 48.9 mmol) were dissolved in dimethylsulfoxide (44 mL) and allowed to stir under nitrogen overnight. The reaction mixture was added to 150 mL of stirring water and diluted with 150 mL ethyl acetate. The organic and aqueous phases were separated. The organic phase was washed with brine (3×100 mL) and dried over magnesium sulfate. After filtering, the organic liquid was concentrated and the resulting solid was dried in vacuo to afford 143 (1.29 g, 55%).

Preparation of Compound 144

Preparation of Compound 145

Compound 71 (2.01 g, 5.98 mmol), allylbromide (2.10 mL, 24.4 mmol), and potassium carbonate (6.61 g, 47.8 mmol) were dissolved in dimethylsulfoxide (44 mL) and stirred overnight. The reaction mixture was added to 150 mL of stirring water and diluted with 150 mL ethyl acetate. The organic and aqueous phases were separated. The organic phase was washed with brine (3×100 mL), dried over magnesium sulfate, filtered, and concentrated. The resulting solid was dried in vacuo to afford 145 (1.98 g, 88%).

Preparation of Compound 146

Preparation of Compound 147

Compound 71 (2.07 g, 6.17 mmol), 2-iodobutane (3.10 mL, 26.9 mmol), and potassium carbonate (6.78 g, 49.1 mmol) were dissolved in dimethylsulfoxide (44 mL) and allowed to stir under nitrogen overnight. The reaction mixture was diluted with ethyl acetate (300 mL). The organic material was washed with water (200 mL) and brine (300 mL) and dried over magnesium sulfate. After filtration, the material was concentrated and the resulting solid was dried in vacuo to afford 147 (1.29 g, 55%).

Preparation of Compound 148

Preparation of Compound 149

Preparation of Compound 150

Preparation of Compound 152

To a solution of 149 (500 mg, 1.03 mmol) in 1,2-dichloroethane (4 mL) was added propionaldehyde (90 μL, 1.24 mmol). After stirring for 10 min, sodium triacetoxyborohydride (306 mg, 1.44 mmol) was added. The reaction mixture stirred under nitrogen for 1.5 h before being quenched with saturated sodium bicarbonate solution (5 mL). The resulting solution was extracted with ethyl acetate (3×7 mL). The organic extracts were combined and dried over sodium sulfate. The organic liquid was filtered and concentrated. Purification by silica gel chromatography (90:10:1 CH2Cl2/methanol/NH4OH) yielded 152.

Preparation of Compound 152 .HCl

Preparation of Compound 151

To a solution of 149 (302 mg, 0.624 mmol) in 1,2-dichloroethane (2.5 mL) was added propionaldehyde (36.0 μL, 0.500 mmol). After stirring for 15 min under nitrogen, sodium triacetoxyborohydride (93.0 mg, 0.874 mmol) was added. After 10 min, another 93.0 mg (0.874 mmol) sodium triacetoxyborohydride were added. The reaction mixture stirred under nitrogen for 1.5 h before being quenched with saturated sodium bicarbonate solution (5 mL). The resulting solution was extracted with ethyl acetate (3×7 mL). The organic extracts were combined and dried over sodium sulfate. The organic liquid was filtered and concentrated. Purification by silica gel chromatography (90:10:1 CH2Cl2/methanol/NH4OH) yielded 151.

Preparation of Compound 151 .HCl

Preparation of Compound 153

To a solution of 61 (2.03 g, 4.44 mmol) in ethylene glycol dimethyl ether (100 mL), was added 2,5-dimethoxyphenylboronic acid (2.42 g, 13.3 mmol), tris(dibenzylideneacetone)dipalladium(0) (0.135 g, 0.148 mmol), triphenylphosphine (0.581 g, 2.22 mmol), and 2 M sodium carbonate solution (10 mL). The dispersion was refluxed overnight under nitrogen. After cooling to room temperature, the reaction mixture was diluted with 100 mL water. The aqueous solution was extracted with methylene chloride (3×100 mL). The extracts were combined and washed with water (300 mL) and brine (300 mL). The organic solution was dried over sodium sulfate, filtered, and concentrated. The product was purified by silica gel chromatography (90:10:1 CH2Cl2/methanol/NH4OH) to yield 153 (900 mg, 39%).

Preparation of 153 .HCl

Preparation of Compound 154

Preparation of Compound 155

Preparation of Compound 155 .HCl

Preparation of Compound 156

Preparation of Compound 156 .HCl

Preparation of Compound 157

To a solution of 61 (1.50 g, 3.27 mmol) in ethylene glycol dimethyl ether (75 mL), was added 5-methyl-2-thiopheneboronic acid (1.40 g, 9.82 mmol), tris(dibenzylideneacetone)dipalladium(0) (100 mg, 0.109 mmol), triphenylphosphine (430 mg, 1.64 mmol), and 2 M sodium carbonate solution (10 mL). The solution was refluxed under nitrogen for 2 d. After cooling to room temperature, the reaction mixture was diluted with 100 mL water. The aqueous solution was extracted with methylene chloride (3×100 mL). The extracts were combined and washed with water (300 mL) and brine (300 mL). The organic solution was dried over sodium sulfate, filtered, and concentrated. The product was purified by silica gel chromatography (90:10:1 CHCl3/methanol/NH4OH) to yield 157 (1.04 g, 67%).

Preparation of Compound 157 .HCl

Preparation of Compound 159

Preparation of Compound 158

Preparation of Compound 160

To a suspension of 61 (1.00 g, 2.18 mmol) in ethylene glycol dimethyl ether (50 mL), was added 4-methylthiophene-2-boronic acid (0.93 g, 6.54 mmol), tris(dibenzylideneacetone)dipalladium(0) (67.0 mg, 0.073 mmol), triphenylphosphine (287 mg, 1.09 mmol), and 2 M sodium carbonate solution (10 mL). The solution was refluxed under nitrogen for 2 d. After cooling to room temperature, the reaction mixture was diluted with 100 mL water. The aqueous solution was extracted with methylene chloride (3×100 mL). The extracts were combined and washed with water (300 mL) and brine (300 mL). The organic solution was dried over sodium sulfate, filtered, and concentrated. The product was purified by silica gel chromatography (90:10:1 CHCl3/methanol/NH4OH) to yield 160 (450 mg, 44%).

Preparation of Compound 160 .HCl

Preparation of Compounds 162 and 163

Preparation of Compound 162 .HCl

Preparation of Compound 163 .HCl

Preparation of Compound 161

Preparation of Compound 164

To a solution of 61 (1.20 g, 2.62 mmol) in ethylene glycol dimethyl ether (75 mL), was added furan-3-boronic acid (0.88 g, 7.85 mmol), tris(dibenzylideneacetone)dipalladium(0) (80.0 mg, 0.087 mmol), triphenylphosphine (343 mg, 1.31 mmol), and 2 M sodium carbonate solution (10 mL). The solution was refluxed under nitrogen overnight. After cooling to room temperature, the reaction mixture was diluted with 100 mL water. The aqueous solution was extracted with methylene chloride (3×150 mL). The extracts were combined and washed with water (450 mL) and brine (450 mL). The organic solution was dried over sodium sulfate, filtered, and concentrated. The product was purified by silica gel chromatography (90:10:1 CHCl3/methanol/NH4OH) to yield 164 (700 mg, 60%).

Preparation of Compound 164 .HCl

Preparation of Compound 166

To a stirred solution of 164 (200 mg, 0.449 mmol) in 1,2-dichloromethane (2 mL) was added propionaldehyde (26.0 μL). After stirring under nitrogen for 20 min, sodium triacetoxyborohydride (114 mg, 0.539 mmol) was added. The mixture was stirred overnight. The reaction was concentrated and purified by silica gel chromatography (90:10:1 CHCl3/methanol/NH4OH) to yield 166.

Preparation of Compound 166 .HCl

Preparation of Compound 165

Preparation of Compounds 167 and 168

Compound 75 (0.500 g, 1.10 mmol) was dissolved in 1,2-dichloroethane (10 mL). To this stirred solution was added acetaldehyde (0.054 g, 1.22 mmol) and sodium triacetoxyborohydride (0.360 g, 1.71 mmol). After 1.5 h, the reaction was quenched with saturated sodium bicarbonate solution (10 mL). The mixture was extracted with ethyl acetate (10 mL). The organic layers were combined, dried over sodium sulfate, and concentrated. The resulting material was purified via silica gel chromatography (60:1:1 CH2Cl2/methanol/triethylamine) to yield 167 (213 mg, 40%), and 168 (109 mg, 19%).

Preparation of Compound 167 .HCl

Preparation of Compound 168 .HCl

Preparation of Compounds 169 and 170

Compound 75 (0.500 g, 1.10 mmol) was dissolved in 1,2-dichloroethane (10 mL). To this stirred solution was added butyraldehyde (0.072 g, 1.00 mmol) and sodium triacetoxyborohydride (0.297 g, 1.40 mmol). After 2.5 h, the reaction was quenched with saturated sodium bicarbonate solution (10 mL). The mixture was extracted with ethyl acetate (10 mL). The organic layers were combined, dried over sodium sulfate, and concentrated. The resulting material was purified via silica gel chromatography (200:10:1 CH2Cl2/methanol/NH4OH) to yield 169 (180 mg, 32%), and 170 (160 mg, 26%).

Preparation of Compound 169 .HCl

Preparation of Compound 171

To a stirred, 0° C. solution of 169 (125 mg, 0.228 mmol) in methylene chloride (10 mL), was added pyridine (46 μL), DMAP (6.0 mg, 0.046 mmol), and acetic anhydride (24.0 μL, 0.251 mmol). After 1 h under a nitrogen atmosphere, the reaction mixture was warmed to room temperature. After stirring overnight, another 2.2 equivalents of acetic anhydride and 0.2 equivalents of DMAP were added and the mixture was heated to reflux. Following concentration, the material was diluted with ethyl acetate (20 mL) and saturated sodium bicarbonate solution (20 mL). The organic layer was concentrated and dried in vacuo. The resulting material was purified via silica gel chromatography (90:10:1 CH2Cl2/methanol/NH4OH) and trituration with hexanes to yield 171 (26 mg): API MS m/z=554 [C33H43N7O+H]+.

Preparation of Compound 170 .HCl

Preparation of Compounds 172 and 173

Compound 75 (0.500 g, 1.10 mmol) was dissolved in 1,2-dichloroethane (10 mL). To this stirred solution was added cyclopropanecarboxaldehyde (0.070 g, 1.00 mmol) and sodium triacetoxyborohydride (0.297 g, 1.40 mmol). After 3 h, the reaction was quenched with saturated sodium bicarbonate solution (10 mL). The mixture was extracted with ethyl acetate (10 mL). The organic layers were combined, dried over sodium sulfate, and concentrated. The resulting material was purified via silica gel chromatography (200:10:1 CH2Cl2/methanol/NH4OH) to yield 172 (103 mg, 18%), and 173 (160 mg, 26%).

Preparation of Compound 172 .HCl

Preparation of Compound 173 .HCl

Preparation of Compounds 174 and 175

Preparation of Compound 267

The HCl salt of 167 (10 mg, 0.037 mmol), was dispersed in ethyl acetate and neutralized with sodium bicarbonate. The organic material was dried over magnesium sulfate and concentrated. The solid was dissolved in dry CH2Cl2(10 mL) and cooled to 0° C. To the solution was added DMAP (9 mg), pyridine (0.074 mL) and acetic anhydride (0.037 mL). The ice bath was removed after 1 h. After being stirred overnight, additional DMAP and Ac2O was added in portions to consume starting material by TLC analysis. The mixture was heated to reflux for 2 d. Upon cooling, the mixture was concentrated in vacuo, then neutralized with aqueous sodium bicarbonate, extracted with ethyl acetate, dried and concentrated. The residue was purified by chromatography to provide 267: API MS m/z=526 [C31H39N7O+H]+.

Preparation of Compound 177

To a solution of 61 (1.00 g, 2.21 mmol) and 3-tolylboronic acid (0.33 g, 2.43 mmol) in tetrahydrofuran (5 mL) was added tris(dibenzylideneacetone)dipalladium(0) (0.010 g, 0.011 mmol), tri-tert-butylphosphine (5.5 mg, 0.027 mmol), and potassium fluoride (0.42 g, 7.29 mmol). After mixing overnight at room temperature, the reaction mixture was refluxed for 24 h and cooled to room temperature. The reaction mixture was diluted with ether (50 mL) and filtered through Celite. The organic liquid was concentrated and the resulting material was purified via silica gel chromatography (90:10:1 CH2Cl2/methanol/NH4OH) to yield 177 (0.70 g, 71%).

Preparation of Compound 177 .HCl

Preparation of Compound 178

To a stirred solution of 177 (219 mg, 0.466 mmol) in methylene chloride (25 mL) was added acetic anhydride (48 mL, 0.513 mmol), DMAP (5.7 mg, 0.047 mmol), and pyridine (57.0 μL, 0.699 mmol). The mixture was placed under a nitrogen atmosphere and immersed in an ice water bath. After 30 min, the reaction mixture was warmed to room temperature and stirred for another 1.5 h. The solution was concentrated and the resulting material was purified via silica gel chromatography (95:5:1 CH2Cl2/methanol/NH4OH) to afford 178.

Preparation of Compound 178 .HCl

Preparation of Compound 179

Preparation of Compound 179 .HCl

Preparation of Compound 180

To a stirred solution of 179 (200 mg, 0.412 mmol) in methylene chloride (25 mL) was added acetic anhydride (43 μL, 0.450 mmol), DMAP (5.0 mg, 0.041 mmol), and pyridine (50.0 μL, 0.618 mmol). The mixture was placed under a nitrogen atmosphere and immersed in an ice water bath. After 30 min, the reaction mixture was warmed to room temperature and stirred for another 1.5 h. The solution was concentrated and the resulting material was purified via silica gel chromatography (95:5:1 CH2Cl2/methanol/NH4OH) to afford 180.

Preparation of Compound 180 .HCl

Preparation of Compound 181

To a solution of 61 (2.00 g, 4.36 mmol) and furan-2-boronic acid (1.50 g, 13.1 mmol) in ethylene glycol dimethyl ether (150 mL) was added tris(dibenzylideneacetone)dipalladium(0) (120 mg, 0.130 mmol), tri-tert-butylphosphine (570 mg, 2.18 mmol), and 2 M sodium carbonate solution (12.5 mL, 25.3 mmol). After refluxing overnight, 2 more equivalents of furan-2-boronic acid were added. The reaction was refluxed for 24 h, cooled to room temperature, and diluted with water (50 mL). The aqueous mixture was extracted with methylene chloride (3×80 mL). The extracts were combined and washed with water (250 mL) and brine (250 mL). The organic phase was dried over sodium sulfate and filtered. The organic liquid was concentrated and the resulting material was purified via silica gel chromatography (95:5:1 CH2Cl2/methanol/NH4OH) to yield 181.

Preparation of Compound 181 .HCl

Preparation of Compound 182

To a stirred solution of 181 (750 mg, 1.68 mmol) in methylene chloride (30 mL) was added acetic anhydride (0.18 mL, 1.85 mmol), DMAP (20.8 mg, 0.17 mmol), and pyridine (0.20 mL, 2.52 mmol). The mixture was placed under a nitrogen atmosphere and immersed in an ice water bath. Afiter 30 min, the reaction mixture was warmed to room temperature and stirred for another 1.5 h. The solution was concentrated and the resulting material was purified via silica gel chromatography (95:5:1 CH2Cl2/methanol/NH4OH) to afford 182 (530 mg, 65%).

Preparation of Compound 182 .HCl

Preparation of Compound 183

To a stirred mixture of 6-chloronicotinamide (5.00 g, 31.9 mmol) in ethanol (13 mL) and toluene (80 mL) was added 3-fluorobenzeneboronic acid (4.92 g, 35.1 mmol) and 2 M sodium carbonate solution (32 mL). The suspension was heated to 80° C. and degassed with argon for 1 h. After cooling to room temperature, tetrakis(triphenylphophine)palladium(0) (1.11 g, 0.958 mmol) was added. The reaction mixture was refluxed under argon for 3 h. After cooling to room temperature, the mixture was diluted with water (100 mL) and filtered. The filter cake was washed with water and dried in vacuo to afford 183 (6.38 g, 92%).

Preparation of Compound 184

To a stirred suspension of 183 (3.00 g, 13.9 mmol) in tetrahydrofuran (25 mL) was added dropwise 1 M borane in THF (97.0 mL, 97.0 mmol). After refluxing for 2 h, the reaction mixture was cooled in an ice bath. The mixture was acidified to pH 1 with 2 N HCl and stirred for 1 h. The pH was raised to a value of 10 by adding 6 N NaOH and the resulting solution was extracted with ethyl acetate (3×50 mL). The extractions were combined, washed with brine (150 mL), and dried over sodium sulfate. The suspension was filtered and concentrated. The resulting material was purified by precipitation as the HCl salt from an ethanol solution. The product was recovered by filtration and dried in vacuo to yield 184 (1.69 g, 51%):1H NMR (300 MHz, CD3OD) δ 8.92 (s, 1 H), 8.41 (d, 1 H), 8.27 (d, 1 H), 7.77-7.90 (m, 2 H), 7.35-7.70 (m, 2 H), 4.36 (s, 2 H).

Preparation of Compound 185

The amine 184 (1.84 g, 7.69 mmol), 2,6-dichloropurine (1.31 g, 6.99 mmol), and N,N-diisopropylethylamine (2.68 mL, 15.4 mmol) were dissolved in ethanol (65 mL). After refluxing overnight, the solution was immersed in an ice water bath for 20 min. The mixture was filtered and cake was washed with water. The cake was triturated with ethanol and diethyl ether and dried in vacuo to afford 185 (1.12 g, 47%).

Preparation of Compound 186

To a stirred solution of 185 (1.00 g, 2.94 mmol) in dimethylsulfoxide (100 mL) was added potassium carbonate (2.19 g, 15.9 mmol) and 2-iodopropane (0.88 mL, 8.81 mmol). The mixture was placed under an argon atmosphere and stirred overnight. The reaction mixture was poured into stirred water (300 mL) and the resulting solution was extracted with ethyl acetate (3×300 mL). The extractions were combined, washed with water (900 mL) and brine (900 mL), and dried over magnesium sulfate. Following filtration, the organic liquid was concentrated. The material was purified by recrystallization from ethyl acetate in hexanes to yield 186 (0.92 g, 82%).

Preparation of Compound 187

Preparation of Compound 188

Preparation of Compound 189

To a stirred mixture of 6-chloronicotinamide (3.00 g, 19.2 mmol) in ethanol (7.6 mL) and toluene (48 mL) were added 3-methoxyphenylboronic acid (3.20 g, 21.1 mmol) and 2 M sodium carbonate solution (19 mL). The suspension was heated to 80° C. and degassed with argon for 1 h. After cooling to room temperature, tetrakis(triphenylphophine)palladium(0) (664 mg, 0.575 mmol) was added. The reaction mixture was refluxed under argon for 3 h. After cooling to room temperature, the mixture was diluted with water (100 mL) and filtered. The filter cake was washed with water and dried in vacuo to afford 189 (3.62 g, 83%).

Preparation of Compound 190

To a stirred solution of 189 (3.00 g, 13.1 mmol) in tetrahydrofuran (25 mL) was added dropwise 1 M borane in THF (92.0 mL, 92.0 mmol). After refluxing for 4 h, the reaction mixture was cooled in an ice bath. The mixture was acidified to pH 1 with 2 N HCl and stirred for 1 h. The pH was raised to a value of 10 by adding 6 N NaOH and the resulting solution was extracted with ethyl acetate (3×50 mL). The extractions were combined, washed with brine (150 mL), and dried over sodium sulfate. The suspension was filtered and concentrated. The resulting material was purified by precipitation as the HCl salt from an ethanol solution. The product was recovered by filtration and dried in vacuo to yield 190 (1.81 g, 55%).

Preparation of Compound 191

The amine 190 (1.80 g, 7.18 mmol), 2,6-dichloropurine (1.22 g, 6.53 mmol), and N,N-diisopropylethylamine (1.86 g, 6.53 mmol) were dissolved in ethanol (82 mL). After refluxing overnight, the dispersion was immersed in an ice water bath for 60 min. The mixture was filtered and cake was washed with water. The cake was triturated with ethanol and diethyl ether and dried in vacuo to afford 191 (1.04 g, 44%).

Preparation of Compound 192

To a stirred solution of 191 (1.04 g, 2.84 mmol) in dimethylsulfoxide (60 mL) was added potassium carbonate (2.12 g, 15.3 mmol) and 2-iodopropane (0.85 mL, 8.52 mmol). The mixture was placed under an argon atmosphere and stirred overnight. The reaction mixture was poured into stirred water (60 mL) and the resulting solution was extracted with ethyl acetate (3×60 mL). The extractions were combined, washed with water (180 mL) and brine (180 mL), and dried over magnesium sulfate. Following filtration, the organic liquid was concentrated. The material was purified by recrystallization from ethyl acetate in hexanes (1:40) to yield 192.

Preparation of Compound 193

In a sealed tube were combined 192 (400 mg, 1.09 mmol), trans-1,4-diaminocyclohexane (1.25 g, 10.9 mmol), and ethanol (4.0 mL). The reaction mixture was heated to 150° C. for 24 h and cooled to room temperature. The solution was filtered and the filtrate was concentrated. Purification by column chromatography (97:3 CH2Cl2/methanol) yielded the free base, 193 (240 mg, 45%).

Preparation of Compound 194

Preparation of Compound 195

To a stirred mixture of 6-chloronicotinamide (2.00 g, 12.8 mmol) in ethanol (5.0 mL) and toluene (32 mL) was added thiophene-2-boronic acid (1.80 g, 14.1 mmol) and 2 M sodium carbonate solution (13 mL). The suspension was heated to 80° C. and degassed with argon for 1 h. After cooling to room temperature, tetrakis(triphenylphophine)palladium(0) (443 mg, 0.383 mmol) was added. The reaction mixture was refluxed under argon for 3 h. After cooling to room temperature, another 0.950 g thiophene-2-boronic acid and 280 mg tetrakis(triphenylphophine)palladium(0) were added to the reaction mixture. It was refluxed for 4 h and cooled to room temperature. The mixture was diluted with water (50 mL) and filtered. The filter cake was washed with water and dried in vacuo to afford 195 (1.65 g, 63%).

Preparation of Compound 196

To a stirred solution of 195 (1.40 g, 6.86 mmol) in tetrahydrofuran (23 mL) was added dropwise 1 M borane in THF (48.0 mL, 48.0 mmol). After refluxing for 1 h, the reaction mixture was cooled in an ice bath. The mixture was acidified to pH 1 with 2 N HCl and stirred for 1 h. The pH was raised to a value of 10 by adding 6 N NaOH and the resulting solution was extracted with ethyl acetate (3×50 mL). The extractions were combined, washed with brine (150 mL), and dried over sodium sulfate. The suspension was filtered and concentrated. The resulting material was purified by precipitation as the HCl salt from an ethanol solution. The product was recovered by filtration and dried in vacuo to yield 196 (0.87 g, 56%).

Preparation of Compound 197

Preparation of Compound 198

To a stirred solution of 197 (200 mg, 0.583 mmol) in dimethylsulfoxide (12 mL) was added potassium carbonate (435 mg, 3.15 mmol) and 2-iodopropane (0.18 mL, 1.75 mmol). The mixture was placed under an argon atmosphere and stirred overnight. The reaction mixture was poured into stirred water (15 mL) and the resulting solution was extracted with ethyl acetate (3×30 mL). The extractions were combined, washed with water (90 mL) and brine (90 mL), and dried over magnesium sulfate. Following filtration, the organic liquid was concentrated to yield 198 (200 mg, 89%).

Preparation of Compound 199

Preparation of Compound 200

Preparation of Compound 201

To a stirred solution of 6-chloronicotinamide (2.80 g, 17.9 mmol) in ethanol (7.5 mL) and toluene (48 mL) was added furan-2-boronic acid (3.00 g, 26.8 mmol) and 2 M sodium carbonate solution (18 mL). The suspension was heated to 80° C. and degassed with argon for 1 h. After cooling to room temperature, tetrakis(triphenylphophine)palladium(0) (619 mg, 0.536 mmol) was added. The reaction mixture was refluxed under argon for 2 d then cooled to room temperature. The mixture was diluted with water (75 mL) and filtered. The filter cake was washed with water and dried in vacuo to afford 201 (1.95 g, 58%).

Preparation of Compound 202

To a stirred solution of 201 (1.69 g, 8.98 mmol) in tetrahydrofuran (34 mL) was added dropwise 1 M borane in THF (50.0 mL, 50.0 mmol). After refluxing for 2 h, the reaction mixture was cooled in an ice bath. The mixture was acidified to pH 1 with 2 N HCl and stirred for 1 h. The pH was raised to a value of 10 by adding 6 N NaOH and the resulting solution was extracted with ethyl acetate (3×50 mL). The extractions were combined, washed with brine (150 mL), and dried over sodium sulfate. The suspension was filtered and concentrated. The resulting material was purified by precipitation as the HCl salt from an ethanol solution. The product was recovered by filtration and dried in vacuo to yield 202 (1.12 g, 50%).

Preparation of Compound 203

Preparation of Compound 204

To a stirred solution of 203 (166 mg, 0.508 mmol) in dimethylsulfoxide (11 mL) was added potassium carbonate (379 mg, 2.74 mmol) and 2-iodopropane (0.150 mL, 1.52 mmol). The mixture was placed under an argon atmosphere and stirred overnight. The reaction mixture was poured into stirred water (15 mL) and the resulting solution was extracted with ethyl acetate (3×30 mL). The extractions were combined, washed with water (90 mL) and brine (90 mL), and dried over magnesium sulfate. Following filtration, the organic liquid was concentrated to yield 204 (178 mg, 95%).

Preparation of Compound 205

In a sealed tube were combined 204 (170 mg, 0.461 mmol), trans-1,4-diaminocyclohexane (526 mg, 4.61 mmol), and ethanol (2.5 mL). The reaction mixture was heated to 150° C. for 4 d and cooled to room temperature. The solution was filtered and concentrated to afford 205.

Preparation of Compound 206

Preparation of Compound 207

Preparation of Compound 208

Preparation of Compound 211

Preparation of Compound 212

Preparation of Compound 213

Preparation of Compound 214

Preparation of Compounds 209 and 210

Preparation of Compound 215

Preparation of Compound 216

Preparation of Compound 217

Prepared by the general methods described above: TLC silica gel Rf=0.52 (20:1:0.01 —CH2Cl2/MeOH/NH4OH).

Preparation of Compound 218

Preparation of Compound 219

Preparation of Compound 221

Preparation of Compound 222

Preparation of Compound 223

Preparation of Compound 224

Preparation of 5-Bromo-2-cyanopyridine

2,5-Dibromopyridine (20.0 g, 84.4 mmol) was dissolved in dimethylformamide (422 mL). To the stirred solution was added copper(I) cyanide. After refluxing for 5 h, the mixture was cooled to room temperature and stored overnight. The reaction mixture was diluted with ethyl acetate (1200 mL) and filtered through a Buchner funnel containing sand, Celite, and silica gel layers. The filtrate was concentrated to a volume of 400 mL. This organic liquid was diluted with water (300 mL) and the resulting liquid was extracted with ethyl acetate (2×200 mL). The organic extracts were combined, washed with water (2×300 mL) and brine (1×250 mL), and dried over magnesium sulfate. After concentration, the product was purified via silica gel chromatography (50:50 ethyl acetate/CH2Cl2) to afford the title compound (9.79 g).

Preparation of Compound 225

Prepared by reation of 5-bromo-2-cyanopyridine with benzeneboronic acid under standard Suzuki conditions (68%).

Preparation of Compound 226

In a Parr shaker vessel were combined 225 (300 mg, 1.67 mmol), glacial acetic acid (25 mL), and 10% palladium on carbon catalyst (177 mg, 0.167 mmol). The solution was agitated under 45 psig hydrogen gas for 2 h. The resulting dispersion was filtered through a Buchner funnel. The filtrate was concentrated. Purification by acid/base extraction yielded 226 (240 mg, 78%).

Preparation of Compound 229

Preparation of Compound 230

Preparation of Compound 231

Preparation of Compound 232

Preparation of Compound 233

Preparation of Compound 239

Preparation of Compound 240

Preparation of Compound 241

Reductive amination of 239 with propionaldehyde followed by salt formation provided 241: ESI MS m/z=512 [C31H41N7+H]+.

Preparation of Compound 242

Preparation of Compound 243

Preparation of Compound 245

To a stirred solution of sodium hydride (423 mg, 17.6 mmol) in tetrahydrofuran (12 mL), was added 4-phenylphenol (2.00 g, 11.8 mmol). After 1 h, BOC-2-aminoethylbromide (3.90 g, 17.6 mmol) was added to the solution. After stirring overnight, the reaction mixture was quenched with 2 N potassium hydroxide solution (10 mL). The resulting mixture was extracted with methylene chloride (12 mL). The organic layer was concentrated and the crude material was purified via silica gel chromatography to yield 245.

Preparation of Compound 246

The protected amine 245 was added to 10 mL of an 1:1 mixture of methylene chloride and trifluoroacetic acid. After concentration, the material was diluted with 2 N potassium hydroxide solution (10 mL). The aqueous layer was extracted with methylene chloride (2×10 mL). The organic extracts were combined, dried over magnesium sulfate, and concentrated in vacuo to afford the product (400 mg). Reaction with 1 under standard conditions provided 246 (91%).

Preparation of Compound 248

Preparation of Compound 250

Reductive amination of 248 with propionaldehyde and salt formation under standard conditions described above provided 250: ESI MS m/z=528 [C31H41N7O+H]+.

Preparation of Compound 249

Preparation of Compound 255

Utilizing reaction conditions described in general above, 251 was converted to 252 (100%). Compound 252 was converted to 253 then 254 and then Boc-protected to make 255 (21%).

Preparation of Compound 256

Compound 255 was treated with phenylboronic acid under standard Suzuki condions. The product was dissolved in methanol and immersed in an ice water bath. Hydrogen chloride gas was bubbled through the solution. The solution was concentrated in vacuo and the resulting material was purified via preparatory HPLC (acetonitrile/water/trifluoroacetic acid) to yield 256 (8 mg).

Preparation of Compound 257

Preparation of Compound 258

Reaction of 75 with propionoyl chloride under standard conditions provides 258 (89%): mp 182-183° C.

Preparation of Compound 259

Reaction of 75 with methyl chloroformate under standard conditions provides 259 (68%): mp 148-150° C.

Preparation of Compound 260

Reaction of 75 with methanesulfonyl chloride under standard conditions provides 260 (56%): mp 143-145° C.

Preparation of Compound 261

Reaction of 75 with cyclopropanecarbonyl chloride under standard conditions provides 261 (87%): mp 196-204° C.

Preparation of Compound 262

Compound 75 (250 mg, 0.549 mmol) and succinic anhydride (60.0 mg, 0.600 mmol) were dissolved in xylene (30 mL). A few drops of dimethylformamide were added to the solution. After refluxing for 48 h, the mixture was concentrated in vacuo. The resulting material was purified via silica gel chromatography (99.5:0.5 CH2Cl2/MeOH) and recrystallized from CH2Cl2in hexanes (1:10) to yield 262 (30.0 mg, 10%): mp 141-147° C.

Preparation of Compound 263

The amine 75 (200 mg, 0.439 mmol) was dissolved in methylene chloride (15 mL). The stirred solution was cooled to −78° C. and N,N-diisopropylethylamine (113 mg, 0.878 mmol) and trifluromethylsulfonylchloride (81.4 mg, 0.483 mmol) were added. After 30 min, the solution was warmed to room temperature. The mixture was cooled to −78° C. and another 1.10 equivalents of trifluoromethylsulfonylchloride and 1.50 equivalents of N,N-diisopropylethylamine were added. After warming to room temperature, the solution was concentrated. The resulting material was purified via silica gel chromatography (99:1 CH2Cl2/MeOH) and recrystallization from ether in hexanes to afford 263 (60 mg, 23%): mp 131-136° C.

Preparation of Compound 264

Prepared by standard Suzuki coupling of 61 to provide 264 (65%): mp 186-190° C.

Preparation of Compound 265

Preparation of Compound 266

Description of Biological Assays

CyclinA/cdk2 and cyclinE/cdk2 assays were carried out with cyclin/cdk complexes isolated from HeLa S-3 suspension cultures. HeLa cells were grown in spinner flasks at 37° C. in Joklik's modified minimum essential media (MEM) supplemented with 7% horse serum. After growing in medium supplemented with 2 mM thymidine for 16-18 h, cultures were arrested at the G1/S border and cyclinA/cdk2 and cyclinE/cdk2 were isolated from cell lysates by immunoprecipitation with antibodies specifically directed against each cyclin subunit. Rabbit anti-cyclinA (H-432) and the mouse monoclonal antibody against cyclinE (HE111) were purchased from Santa Cruz Biotechnology. Cells blocked at the appropriate stage of the cell cycle were disrupted in lysis buffer (50 mM Tris, pH 8.0, 250 mM NaCl, 0.5% NP-40 plus protease and phosphatase inhibitors) and centrifuged at 10,000×g to remove insoluble material. To isolate cyclin/cdk complexes, 1 μg of anti-cyclin antibody was incubated with lysate from 1×107cells for 1 h at 4° C. Protein A-coated agarose beads were then added for 1 h to collect antibody-bound immune complexes. The immobilized cyclin/cdk complexes were then washed 4× with lysis buffer to reduce nonspecific protein binding. The complexes were then washed 1× in kinase assay buffer (50 mM Tris-HCl, pH 7.4, 10 mM MgCl2, 1 mM DTT) and aliquoted into individual assay tubes.

HeLa cells are blocked at the G1/S border by culturing in the presence of 2 mM thymidine for 20 h. The cells are then rinsed 3× in phosphate buffered saline and resuspended in regular medium. After 4 h of culture, the mitotic blocker, nocodazole is added to a final concentration of 75 ng/ml. Sixteen hours later, the cells are harvested by centrifugation, washed in PBS, and lysed in cold Lysis Buffer (50 mM Tris pH 8.0, 250 mM NaCl, 0.5% NP-40, 1 mM DTT, 25 μg/ml leupeptin, 25 μg/ml aprotinin, 15 μg/ml benzamidine, 1 mM PMSF, 50 mM sodium fluoride, 1 mM sodium orthovanadate) for 15 min at 1×107cells/ml. The lysate is then clarified by centrifugation at 10,000×g for 10 min. The supernatant is collected and diluted 1:5 with Lysis Buffer. Monoclonal antibody against cyclinB (GNS1) is added to the supernatant to a final concentration of 5 μg/ml and shaken at 4° C. for 2 h. The immune complexes are then collected by the addition of 200 μl of protein agarose beads for 1 h. The beads are washed 4× in lysis buffer and 1× in kinase assay buffer.

C. Protein Kinase Assays and Determination of IC50Values

CyclinA/cdk2 assays were carried out with complexes isolated from 0.5×106cells. CyclinE/cdk2 assays were carried out with complexes isolated from 4×106cells. CyclinB/cdk1 assays were carried out with complexes isolated from 4×104cells. After centrifugation, the wash buffer was removed and the complexes resuspended in 15 μl of kinase assay buffer (kinase wash buffer+167 μg/ml histone H1). Compounds being tested for inhibition were added prior to the addition of [γ32P] ATP to a final concentration of 15 μM. The tubes were incubated at 30° C. for 5 min and the reactions were stopped by the addition of an equal volume of 2×SDS-PAGE sample buffer. The samples were then subjected to electrophoresis on 10% SDS-PAGE to resolve the histone H1 from other reaction components. The amount of radioactive phosphate transferred to histone H1 was quantified on a Storm Phosphorimager (Molecular Dynamics).

Prior to the protein kinase assay, test compounds were dissolved in DMSO at a concentration of 25 mM and were diluted to produce final concentrations of 0.1, 1.0, and 10.0 μM in the kinase assays. To eliminate possible effects of differences in DMSO concentration, the DMSO was kept constant at 0.04%, including the control reaction. Duplicate assays were performed at each concentration. The activity was plotted as the percent of activity in the absence of added test compound versus test compound concentration. IC50values were calculated using GraphPad Prism data analysis software.

D. Measuring the Inhibition of Cell Growth

Growth inhibition (GI50) values were measured with HeLa S-3 cells selected for growth on plastic. The procedure was based on the protocol of Skehan et al. (Skehan, P., et al.,J. Natl. Cancer Inst.,82:1107-1112 (1990), which is hereby incorporated by reference) HeLa cells were plated at 2×104cells/well in 96 well plates. One day later, a control plate was fixed by addition of TCA to 5%. After five rinses with tap water the plate was air dried and stored at 4° C. Test compounds were added to the remaining plates at 10-fold dilutions between 0.01 and 100 μM. Two days later all plates were fixed as described above. Cells were then stained by the addition of 100 μl per well of 0.4% sulforhodamine B (SRB) in 1% acetic acid for 30 min at 4° C. Wells were then quickly rinsed 5× with acetic acid (1%) and allowed to air dry. The SRB was then solubilized by the addition of 100 μl per well of unbuffered 10 mM Tris base. Dye was quantified by measuring absorbance at 490 nm on a Molecular Devices kinetic microplate reader. Growth at each inhibitor concentration relative to the untreated control was calculated according to the following equation: percent growth=100×(T−To)/(C−To), where T was the average optical density (OD) of the test wells after 2 days of treatment, Towas the average OD of the wells in the control plate on day 0 and C was the average OD of untreated wells. Plots of percent growth versus inhibitor concentration were used to determine the GI50.

The data below shown in Table 2 summarizes the in vitro cyclin/cdk inhibition constants (IC50) and growth inhibition constants (GI50) of HeLa Cells for the compounds of the current invention. Replicate experimental results are summarized below.

TABLE 2In Vitro Cyclin/cdk Inhibition (IC50) and Growth Inhibition (GI50) of HeLaCells For Compounds of the Current Invention.IC50CyclinA/cdk2IC50CyclinE/cdk2IC50CyclinB/cdk1GI50HeLa CellsCompound(μM)(μM)(μM)(μM)5>1012750.40.6>10122130.060.730.0030.90.50.0010.20.10.020.000113424310.320.80.91430.470.4320.030.031711100.420.930.610.2110.25>1090.41020.30.42514>10261>100.4>109>13223—550.90.733>104>101136280.934125>1071326736>10>10>1020>10>1020>10>1038>10>10>100.6>10>1010.640>10>10>109>10>1025>1043>10>10>104>10>104846>106>102583>1048221>100.3650.60.550>10>10>10379>1053>1015>100.2>1040.30.558112122440.50.760>1012>1070.4>10673>504>100.314120.5>10>100.3>10>100.5745260.2230.01120.050.030.05753360.090.020.0057612360.071150.01320.060.20.0477>104>100.15>10140.50.3780.90.60.80.050.90.30.80.0250.70.20.080.0027910230.070.50.10.00710.080.0040.480>10>10>10>100>104>102860.90.420.20.70.20.030.40.40.010.60.030.010.2874150.0720.30.010.50.10.0040.0060.030.0060.0010.00018834>100.1>10>100.05250.040.005930.20.090.90.30.30.10.080.3940.60.30.40.10.20.30.070.4951140.0820.70.0030.0005968460.040.0197>1031039862>10>10221199>109>105100>104>100.610131410.90.71102>104—41030.60.210.030.70.20.0080.020.011047120.4810.2106113—0.3410.110712—0.440.310810>10—3>10>1051090.60.1—0.04<0.00011100.62—0.020.030.020.011110.20.07—0.020.000611222—<0.0010.0020.020.0060.00061130.40.3—<0.0010.000010.030.0010.0211430.7—>1011530.4—3116>10>10—>10>10117>103—311861—>10>101230.20.04—<0.001<0.0010.000112420.8—0.003<0.001<0.0001130———>10>10131———32132———43133———>10>10134———23135———43137———0.050.060.05139———0.20.07140———12142———0.40.5144———0.40.4146———0.70.3148———11149———0.30.2150———0.30.2151———0.80.6152———0.70.3153———32154———0.60.9155———0.50.8156———32157———0.40.5158———0.60.4159———43160———0.20.3161———0.20.4162———0.20.3163———23164———0.20.1165———0.20.1166———42167———20.9168———43169———0.50.3170———42171———33172———0.30.3173———33174———0.040.030.10.060.40.4175———0.60.3177———0.20.060.06178———0.40.2179———0.10.050.05180———0.40.3181———0.04182———0.30.3187———0.050.03188———0.20.07194———0.060.04199———0.20.09200———0.30.2206———0.20.2207———0.40.2208———43209———22210———34211———0.60.3212———53213———32214———55215———23216———0.50.5217———44218———35219———0.40.6221———22222———12223———0.040.1224———22229———0.4230———0.3231———0.04232———0.3233———0.5239———46240———88241———74242———7>10243———33248———34249———>10>10250———36256———43257———33258———0.20.30.4259———0.30.40.7260———0.20.10.2261———0.30.30.3262———0.30.20.5263———234264———0.30.30.5265———0.30.30.4266———0.30.30.5267———0.80.6

The data below shown in Table 3 summarizes the in vitro cyclin/cdk inhibition (IC50) and growth inhibition (GI50) of HeLa Cells for several reference compounds in comparison to several compounds of the current invention. The chemical structures are provided.

TABLE 7In Vitro Growth Inhibition (GI50) of NCI Human Transformed Cell Linesof Several Compounds of the Current Invention and Olomoucine.Olomoucine GI50Cancer TypeCell Line74 GI50(μM)78 GI50(μM)77 GI50(μM)(μM)BreastBT-5490.160.04<0.0179BreastHS 578T<0.01—<0.0163BreastMCF7<0.01<0.010.0350BreastMDA-MB-231/ATCC<0.01<0.010.04100BreastMDA-MB-435———63BreastMDA-N<0.01<0.010.0179BreastNCI/ADR-RES0.2414.450.03100BreastT-47D<0.010.030.0163CNSSF-268<0.01—<0.0150CNSSF-295<0.010.210.0479CNSSF-5390.07—0.2232CNSSNB-19<0.01<0.010.0363CNSSNB-75<0.01<0.01<0.0125CNSU251<0.010.020.0950ColonCOLO 205<0.01<0.010.0232ColonHCC-2998—<0.01—63ColonHCT-116<0.010.030.0540ColonHCT-15<0.011.48<0.0140ColonHT29<0.01<0.01<0.0163ColonKM12<0.01<0.01<0.0140ColonSW-620<0.01<0.01<0.0140LeukemiaCCRF-CEM<0.01—<0.0140LeukemiaHL-60(TB)—<0.01—40LeukemiaK-562<0.010.020.02100LeukemiaMOLT-4<0.01<0.010.0163LeukemiaRPMI-8226<0.01<0.01<0.0150LeukemiaSR<0.01—0.0225MelanomaLOX IMVI<0.01—0.0432MelanomaM14<0.01<0.01<0.01100MelanomaMALME-3M0.010.010.05100MelanomaSK-MEL-20.060.020.51100MelanomaSK-MEL-28<0.010.01<0.0150MelanomaSK-MEL-50.060.100.0840MelanomaUACC-257<0.010.020.0279MelanomaUACC-620.040.030.1232Non-Small CellA549/ATCC<0.01<0.01<0.0150LungNon-Small CellEKVX—0.05—100LungNon-Small CellHOP-62<0.010.02<0.0132LungNon-Small CellHOP-920.03—0.1350LungNon-Small CellNCI-H226—0.02—50LungNon-Small CellNCI-H23<0.010.010.0179LungNon-Small CellNCI-H322M<0.01<0.01<0.0163LungNon-Small CellNCI-H460<0.010.050.2263LungNon-Small CellNCI-H522<0.01<0.01<0.0140LungOvarianIGROV1<0.01<0.010.0940OvarianOVCAR-3<0.010.030.0279OvarianOVCAR-4<0.010.02<0.01100OvarianOVCAR-50.03<0.010.0440OvarianOVCAR-8<0.010.020.0263OvarianSK-OV-30.220.060.19100ProstateDU-1450.020.060.1340ProstatePC-3<0.01<0.010.02100Renal786-0<0.010.040.0363RenalA4980.030.030.0332RenalACHN0.030.320.1125RenalCAKI-1—0.79—32RenalRXF 393<0.01<0.01<0.0120RenalSN12C<0.01<0.01<0.01100RenalTK-10<0.010.070.0563RenalUO-310.010.17<0.0132
The following data in Table 8 summarize the in vivo properties of several compounds of the current invention. These data were cooperatively obtained at the National Cancer Institute in their Hollow Fiber Assay according to published procedures (Hollingshead, M. G., et al “In Vivo Cultivation of Tumor Cells in Hollow Fibers,”Life Sciences,1995, 57(2), 131-141 which is hereby incorporated by reference).

The following data in Table 9 summarize the in vivo properties of several compounds of the current invention. The protocol for the experiment is as follows. The dose-range finding study consists of four groups of three athymic mice each (four dose levels). The compound is administered on the basis of individual animal body weight. The route is intraperitoneal (IP) and the treatment schedule is daily for 14 days (qd×14) or once every 4 days for 12 days (q4d×3). The mice were observed for survival, and body weights recorded weekly.

The efficacy study consists of three compound-treated groups (six mice/group), a positive control-treated group (six mice), and a vehicle-treated control group of 12 mice. Test compounds were administered IP under the treatment schedules listed above (qd×14 or q4d×3), whereas the positive control agent (Taxol) was administered intravenously (IV) at a dosage level of 15 mg/kg/dose for five consecutive days (qd×5). All agents were administered on the basis of individual animal body weight. Treatment began when the implanted tumors were approximately 100 mg in size (range of 65 to 200 mg). The mice were observed daily for survival. Each tumor was measured by caliper in two dimensions and converted to tumor mass using the formula for a prolate ellipsoid (a×b2/2) and assuming unit density. Tumor measurements and animal body weights were recorded twice weekly. Antitumor activity was assessed by the delay in tumor growth of the treated groups in comparison to the vehicle-treated control group, partial and complete regressions, and tumor-free survivors.