2,4-Pyrimidinediamine Compounds And Uses As Anti-Proliferative Agents

1 . A method of inhibiting proliferation of a cancer cell, comprising contacting the cancer cell with an effective amount of a 2,4-pyrimidinediamine compound according to structural formula (I):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein:

L 1 and L 2 are each, independently of one another, selected from a lower alkyldiyl linker, a lower allylene linker and a covalent bond;

R 2 is selected from the group consisting of lower alkyl optionally substituted with an R b group,

 where Y is NH, O or CH 2 ;

R 2′ is hydrogen, methyl or lower alkyl;

R 4′ is hydrogen, methyl or lower alkyl;

R 4 is selected from the group consisting of lower alkyl optionally monosubstituted with an R a or R b group, lower cycloalkyl optionally monosubstituted with an R a or R b group, lower cycloheteroalkyl optionally substituted at one or more ring carbon and/or heteroatoms with an R a or R b group, —(CR a R a ) n —R b ,

 where D is —(CR 7 R 7 ) m —,

 where Z 1 is N or CH and Z 2 is O, S, NH, S(O) or S(O) 2 ;

R 5 is selected from the group consisting of halo, fluoro and —CF 3 ;

R 6 is hydrogen;

each R 7 is independently selected from the group consisting of hydrogen, methyl, lower alkyl and halo;

each R 8 is independently selected from the group consisting of hydrogen, lower alkyl, —(CH 2 ) n —OH, —OR a , —(CH 2 ) n —NR c R c , —O(CH 2 ) n —R a , —O(CH 2 ) n —R b , —C(O)OR b , —C(S)OR a halo, —CF 3 and —OCF 3 ;

each R 9 is independently selected from the group consisting of hydrogen, lower alkyl, —OR a , —(CH 2 ) n —NR c R c , —O(CH 2 ) n —R a , —O(CH 2 ) n —R b , —C(O)—NR c R c , —C(S)—NR c R c , —S(O) 2 NR c R c , —NHC(O)R a , —NHC(S)R a , —C(O)—NH—(CH 2 ) n —NR c R c , —C(S)—NH—(CH 2 ) n —NR c R c , halo, —CF 3 , —OCF 3 ,

each R 10 is independently selected from the group consisting of hydrogen, lower alkyl, —(CH 2 ) n —OH, —(CH 2 ) n —NR c R c , —OR a , —O(CH 2 ) n —R a , O(CH 2 ) n —R b , halo, —CF 3 , —OCF 3

each R 11 is independently selected from the group consisting of —OR a , —NR c R c and —NR a R d ;

each R 12 is independently selected from the group consisting of lower alkyl, arylalkyl, —OR a , —NR c R c , —C(O)R a , —C(O)OR a and —C(O)NR c R c ;

each R 13 is independently selected from the group consisting of lower alkyl, hydroxy, lower alkoxy, methoxy, —C(O)NR c R c and —C(O)NH 2 ;

each R 15 is independently selected from the group consisting of hydrogen, lower alkyl, lower cycloakyl and phenyl;

each R 16 is independently selected from the group consisting of hydrogen, methyl, lower alkyl, lower cycloalkyl, lower branched alkyl and lower cycloalkylmethyl;

each R 17 is independently selected from the group consisting of hydrogen, lower alkyl, methyl and R d or, alternatively, R 17 may be taken together with R 18 to form an oxo (═O) group;

each R 18 is independently selected from the group consisting of hydrogen, lower alkyl and methyl or, alternatively, R 18 may be taken together with R 17 to form all oxo (═O) group;

each R 19 is independently selected form the group consisting of hydrogen, lower alkyl, methyl and R d ;

each R 20 is independently selected from the group consisting of hydrogen, lower alkyl, methyl and R d ;

each m is independently an integer from 1 to 3;

each n is independently an integer from 1 to 3;

each R a is independently selected from the group consisting of hydrogen, lower alkyl, lower cycloalkyl, lower cycloalkylalkyl, phenyl and benzyl;

each R b is independently selected from the group consisting of —OR a , —CF 3 , —OCF 3 , NR c R c , C(O)R a , —C(S)R a , —C(O)OR a , —C(S)OR a , —C(O)NR c R c , —C(S)NR c R c , —S(O) 2 NR c R c , —C(O)NR a R d , —C(S)NR a R d and —S(O) 2 NR a R d ;

each R c is independently selected from the group consisting of hydrogen, lower alkyl and lower cycloalkyl, or, alternatively, two R c s may be taken together with the nitrogen atom to which they are bonded to form a 5-7 membered saturated ring which optionally includes 1-2 additional heteroatomic groups selected from O, NR a , NR a —C(O)R a , NR a —C(O)OR a and NR a —C(O)NR 8 ; and

each R d is independently selected from lower mono-hydroxyalkyl and lower di-hydroxyalkyl,

with the provisos that:

(i) when R 2 is

 then R 9 and R 10 are not both simultaneously lower alkoxy or methoxy or when R 2 is 3,4,5-trimethoxyphenyl or 3,4,5-tri(loweralkoxy)phenyl, then R 4 is

(ii) when R 2 is lower alkyl, then R 4 is

(iii) when R 4 is

 and L 2 is lower alkylene, then R 2 is other than 3-(1,3-oxazolyl)phenyl;

(iv) when R is

 where R 15 is t-butyl and R 2 is

 then at least two of R 8 , R 9 and R 10 or R 8 , R 9 and R 13 are other than hydrogen;

(v) when R 4 is

 where R 15 is t-butyl and R 2 is

 where R 8 and R 9 are each hydrogen, then R 10 is other than —(CH 2 ) n —OH or —O(CH 2 ) n —R b where R b is selected from —N c R c , —C(O)R a , —C(O)NR c R c and —C(O)NR a R d .

2 . The method of claim 1 in which L 1 and L 2 are each a covalent bond.

3 . The method of claim 2 in which R 5 is fluoro.

4 . The method of claim 3 in which R 2′ and R 41 are each hydrogen.

5 . The method of claim 3 in which R 2′ is hydrogen and R 4′ is methyl.

6 . The method of claim 3 in which R 2 is

7 . The method of claim 6 in which R 4 is selected from unsubstituted cyclopropyl, unsubstituted cyclobutyl, unsubstituted cyclopentyl, unsubstituted cyclohexyl

where R e and R f are selected from (C1-C3) alkanyl and methyl and R g is benzyl.

8 . The method of claim 7 in which R 8 is hydrogen; R 9 is selected from

and R 10 is other than

9 . The method of claim 8 in which R 10 is selected from hydrogen, methyl, methoxy, hydroxymethyl, trifluoromethyl and chloro.

10 . The method of claim 8 in which R 12 is selected from methyl, —C(O)CH 3 , —C(O)OCH 3 and —C(O)OCH 2 CH 3 .

11 . The method of claim 8 in which R 8 is hydrogen; R 9 is other than

and R 10 is selected from

12 . The method of claim 11 in which R 9 is selected from hydrogen, methyl, methoxy, hydroxymethyl, trifluoromethyl and chloro.

13 . The method of claim 11 in which R 12 is selected from methyl, —C(O)CH 3 , —C(O)OCH 3 and —C(O)CH 2 CH 3 .

14 . The method of claim 8 in which R 9 is other than

and R 10 is other than

15 . The method of claim 14 in which R 8 and R 9 are each hydrogen and R 10 is —OCH 2 NHR a .

16 . The method of claim 14 in which R 8 , R 9 and R 10 are each, independently of one another, selected from hydrogen, methyl, methoxy, hydroxymethyl, trifluoromethyl and chloro, with the proviso that at least two of R 8 , R 9 and R 10 are other than hydrogen.

17 . The method of claim 7 in which the 2,4-pyrimidinediamine compound is selected from any compound in any one of TABLEs 1A-1D having an IC 50 of ≦20 μM against at least one tumor cell line in an in vitro antiproliferation assay.

18 . The method of claim 3 in which R 4 is

19 . The method of claim 18 in which R a is hydrogen and R 15 is selected from lower branched alkyl and lower cycloalkyl.

20 . The method of claim 18 in which R 15 is t-butyl or cyclopropyl.

21 . The method of claim 18 in which R 2 is

22 . The method of claim 21 in which R 8 is hydrogen; R 9 is selected from

and R 10 is other than

23 . The method of claim 22 in which R 10 is selected from methyl, trifluoromethyl and chloro.

24 . The method of claim 22 in which R 12 of R 9 is selected from methyl, —C(O)CH 3 , —C(O)OCH 3 and —C(O)CH 2 CH 3 .

25 . The method of claim 21 in which R 8 is hydrogen; R 9 is other than

and R 10 is selected from

26 . The method of claim 25 in winch R 9 is selected from hydrogen, methyl, trifluoromethyl and chloro.

27 . The method of claim 25 in which R 12 of R 10 is selected from methyl, —C(O)CH 3 , —C(O)OCH 3 and —C(O)CH 2 CH 3 .

28 . The method of claim 18 in which the 2,4-pyrimidinediamine compound is selected from any compound in TABLE 2 having an IC 50 of ≦20 μM against at least one tumor cell line in an in vitro antiproliferation assay.

29 . The method of claim 3 in which R 4 is lower alkyl.

30 . The method of claim 29 in which the lower alkyl is branched.

31 . The method of claim 30 in which R 4 is i-propyl or t-butyl.

32 . The method of claim 30 in which R 2 is

33 . The method of claim 32 in which R 8 is hydrogen; R 9 is selected from

and R 10 is other than

34 . The method of claim 33 in which R 10 is selected from hydrogen, methyl, trifluoromethyl and chloro.

35 . The method of claim 33 in which R 2 of R 9 is methyl.

36 . The method of claim 32 in which R 8 is hydrogen; R 9 is hydrogen and R 10 is selected from

37 . The method of claim 36 in which R 12 is methyl.

38 . The method of claim 18 in which the 2,4-pyrimidinediamine compound is selected from any compound in TABLE 3 having an IC 50 of ≦20 μM against at least one tumor cell line in an in vitro antiproliferation assay.

39 . The method of claim 3 in which R 4 is selected from

40 . The method of claim 39 in which R 9 and R 20 are different from one another such that the carbon atom to which they are bonded is chiral.

41 . The method of claim 40 in which the 2,4-pyrimidinediamine compound is a racemate of R and S enantiomers.

42 . The method of claim 40 which the 2,4-pyrimidinediamine compound is enriched in the R enantiomer.

43 . The method of claim 42 in which the 2,4-pyrimidinediamine compound is substantially free of the S enantiomer.

44 . The method of claim 40 in which the 2,4-pyrimidinediamine compound is enriched in the S enantiomer.

45 . The method of claim 44 in which the 2,4-pyrimidinediamine compound is substantially free of the R enantiomer.

46 . The method of claim 39 in which R 2 is

47 . The method of claim 46 in which R 8 , R 9 and R 10 are each, independently of one another, selected from hydrogen, lower alkyl, methyl, lower alkoxy, methoxy, halo, chloro and OCH 2 C(O)NHR a , with the proviso that at least one of R 8 , R 9 or R 10 is other than hydrogen.

48 . The method of claim 46 in which R 8 is hydrogen; R 9 is selected from

and R 10 is selected from hydrogen, methyl and chloro.

49 . The method of claim 48 in which R 12 is selected from methyl, —C(O)CH 3 , —C(O)OCH 3 and —C(O)OCH 2 CH 3 .

50 . The method of claim 46 in which R 8 is hydrogen, R 9 is selected from hydrogen, methyl and chloro; and R 10 is selected from

51 . The method of claim 50 in which R 12 is selected from methyl, —C(O)CH 3 , —C(O)OCH 3 and —C(O)OCH 2 CH 3 .

52 . The method of claim 46 in which the stubstituents on the R 4 and R 2 groups are independently selected from any combinations described in TABLE 4 or TABLE 6.

53 . The method of claim 46 in which the 2,4-pyrimidinediamine compound is selected from any compound in TABLE 4 or TABLE 6 having an IC 50 of ≦20 μM against at least one tumor cell line in an in vitro antiproliferation assay.

54 . The method of claim 3 in which R 4 is

55 . The method of claim 54 in which D is selected from —CH 2 —CH 2 —, —CF 2 —CF 2 , —CH 2 —. and —CF 2 —.

56 . The method of claim 54 in which R 2 is

57 . The method of claim 56 in which R 8 , R 9 and R 10 are each, independently of one another, selected from hydrogen, lower alkyl, methyl, hydroxy, lower alkoxy, methoxy, halo, chloro, fluoro, —OCH 2 C(O)NHR a , —OCH 2 C(O)NR c R c , —OCH 2 C(O)OR a , trifluoromethyl and —O(CH 2 ) n OH, with the proviso that at least one of R 8 , R 9 or R 10 is other than hydrogen.

58 . The method of claim 54 which is selected from any compound in TABLE 5 having an IC 50 of ≦20 μM against at least one tumor cell line in an in vitro antiproliferation assay.

59 . The method of claim 3 in which R 2 is

60 . The method of claim 59 in which R 11 is selected from any R 11 group listed in TABLE 7.

61 . The method of claim 59 in which the 2,4-pyrimidinediamine compound is selected from any compound in TABLE 7 having an IC 50 of ≦20 μM against at least one tumor cell line in an in vitro antiproliferation assay.

62 . The method of claim 3 in which R 4 is

63 . The method of claim 62 in which R 2 is

64 . The method of claim 63 in which each R 8 , R 9 and R 10 is selected such that the compound is not an N2,N4-bis(substituted phenyl)-2,4-pyrimidiniediamine.

65 . The method of claim 63 in which each R 8 , R 9 and R 10 is, independently of the others, selected from hydrogen, lower alkyl, methyl, hydroxy, trifluoromethoxy, lower linear alkoxy, halo, —O(CH 2 ) n NR c R c , —OCH 2 C(O)NHR a , —OCH 2 C(O)NR c R c and —OCH 2 C(O)OR a .

66 . The method of claim 65 in which R 8 , R 9 and R 10 are each, independently of one another; selected from hydrogen, methyl, trifluoromethyl, methoxy, trifluoromethoxy, and chloro, with the proviso that at least one of R 8 , R 9 and R 10 on both of R 2 and R 4 is other than hydrogen.

67 . The method of claim 66 in which at least two of R 8 , R 9 and R 10 on both of R 2 and R 4 are other than hydrogen.

68 . The method of claim 63 in which R 8 and R 9 of R 2 are each hydrogen; R 10 of R 2 is —OCH 2 C(O)NR c R c or —OCH 2 C(O)NHR a ; R 8 and R 10 of R 4 are, independently of one another, selected from hydrogen, methyl and chloro; and R 9 of R 4 is selected from hydroxy,

69 . The method of claim 63 in which the 2,4-pyrimidinediamine compound is selected from any compound in TABLE 8 having an IC 50 of ≦20 μM against at least one tumor cell line in an in vitro antiproliferation assay.

70 . The method of claim 3 in which R 2 is

71 . The method of claim 70 in which R 4 is

where D is —CH 2 CH 2 —.

72 . The method of claim 71 in which R 8 , R 9 and R 10 are each, independently of one another, selected from hydrogen, methyl, methoxy, fluoro and chloro.

73 . The method of claim 71 which is selected from any compound in TABLE 10 having an IC 50 of ≦20 μM against at least one tumor cell line in an in vitro antiproliferation assay.

74 . The method of claim 3 in which R 2 is

75 . The method of claim 74 in which R 4 is

76 . The method of claim 75 in which R 16 is selected from hydrogen, lower linear alkanyl, lower branched alkanyl, lower cyclic alkanyl and lower cyclicalkanylmethyl.

77 . The method of claim 75 in which the 2,4-pyrimidinediamine compound is selected from any compound in TABLE 13 having an IC 50 of ≦20 μM against at least one tumor cell line in an in vitro antiproliferation assay.

78 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is selected from any compound illustrated in TABLES 1-14 having an IC 50 of ≦20 μM against at least one tumor cell line in an in vitro antiproliferation assay.

79 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is a compound according to structural formula (Ia):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein:

m is as integer ranging from 1 to 3;

R 21 is selected from hydrogen, lower alkyl, methyl, lower alkoxy, methoxy, halogen and chloro;

R 22 is selected from hydrogen, lower alkoxy, methoxy, halogen, chloro,

R 23 is selected from hydrogen, methyl, lower alkyl, lower alkoxy, methoxy, halogen, chloro, trifluomethyl, —OCH 2 C(O)NHR a ,

 and

R 24 is selected from hydrogen and methyl,

with the provisos that (i) when R 22 is

 then R 23 is other than

 and (ii) when R 23 is

 then R 22 is other than

80 . The method of claim 79 in which R 21 and R 22 are each hydrogen and R 23 is selected from —OCH 2 C(O)NHR a ,

81 . The method of claim 79 in which R 22 is selected from hydrogen methoxy, chloro,

82 . The method of claim 81 in which R 12 is methyl.

83 . The method of claim 79 in which R 21 is hydrogen and R 22 is

where R 12 is methyl.

84 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is a compound according to structural formula (Ib):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein:

R 15 is selected from t-butyl and cyclopropyl;

R 21 , R 22 and R 23 are as defined in claim 79 .

85 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is a compound according to structural formula (Ic):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein:

R 14 is selected from i-propyl and t-butyl; and

R 21 , R 22 and R 23 are as defined in claim 79 .

86 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is a compound according to structural formula (Id):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein:

Z 1 is selected from N and CH;

Z 2 is selected from NH, O, S and S(O) 2 ;

R 16 is selected from hydrogen and methyl;

R 19 is as defined in claim 1;

R 20 is as defined in claim 1;

R 24 is as defined in claim 79;

R 41 is selected from hydrogen, lower alkyl, methyl; hydroxy, lower alkoxy, methoxy, halo, chloro, trifluoromethyl and —CH 2 OH;

R 42 is selected from hydrogen, lower alkyl, methyl, hydroxy, lower alkoxy, methoxy, halo, chloro, trifluoromethyl, trifluoromethoxy, —OCH 2 C(O)NHR a ,

 where R 12 is as defined in claim 1; and

R 43 is selected from hydrogen, lower alkyl, methyl, hydroxy, lower alkoxy, methoxy, halo, chloro, trifluoromethyl, trifluoromethoxy, —OCH 2 C(O)NHR a , —OCH 2 C(O)OR a ,

 where R 12 is as defined in claim 1 ,

with the provisos that:

(i) when R 42 is

 then R 43 is other than

 and

(ii) when R 43 is

 then R 42 is other than

87 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is a compound according to structural formula (Ie):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein Z 1 , Z 2 , R 16 , R 19 , R 20 , R 24 , R 41 , R 42 and R 43 are as defined in claim 86 .

88 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is a compound according to structural formula (If):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein:

Z 3 is selected from N and CH;

p is 0 or 1;

R 7 is selected from hydrogen and fluoro;

R 24 is as defined in claim 79;

R 51 is selected from hydrogen, lower alkyl, methyl, lower alkoxy, methoxy, halo, trifluoromethyl, CH 2 OH and —C(O)OCH 3 ;

R 52 is selected from hydrogen, lower alkyl, methyl, hydroxy, lower alkoxy, methoxy, halo, trifluoromethyl, CH 2 OH and —OCH 2 C(O)NR a R a where R a is as defined in claim 1; and

R 53 is selected from hydrogen, lower alkyl, methyl, lower alkoxy, methoxy, halo, trifluoromethyl, CH 2 OH, —OCH 2 C(O)OR a and —OCH 2 C(O)NHR a where R a is as defined in claim 1 .

89 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is a compound according to structural formula (Ig):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein:

R 28 is hydrogen or methyl;

R 16 is as defined in claim 86; and

R 21 , R 22 , R 23 and R 24 are as defined in claim 79 .

90 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is a compound according to structural formula (Ih):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein:

R 24 is as defined in claim 79;

R 61 is selected from H and Cl;

R 62 is selected from —CH 2 NHBoc, —CH 2 NH 2 , —C(O)NH 2 , —C(O)NH—CH 2 —C(O)OH, —C(O)NH—CH 2 —C(O)OMe and —C(O)NH—CH 2 CH 2 —N(Et) 2 ;

R 63 is selected from H and Cl;

R 71 is H;

R 72 is selected from H, OMe, Cl and —C(O)NH 2 ; and

R 73 is selected from H, OMe and Cl.

91 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is a compound according to structural formula (Ii):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein:

each R 24 is independently as defined in claim 79; and

R 30 is selected from alkanyl and alkenyl.

92 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is a compound according to structural formula (Ij):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein:

R 24 is as defined in claim 79;

R 81 is selected from hydrogen, methyl and methoxy;

R 82 is selected from hydrogen, methoxy and chloro; and

R 83 is selected from methyl, methoxy, halo and fluoro.

93 . The method of claim 1 in which the 2,4-pyrimidinediamine compound is a compound according to structural formula (Ik), (Il), (Im) or (In):

including prodrugs, salts, hydrates, solvates and N-oxides thereof, wherein:

R 91 , if present, is hydrogen;

R 92 , if present, is selected from lower alkoxy, methoxy, halo, chloro and fluoro;

R 93 , if present, is selected from lower alkoxy, methoxy, halo and chloro; and

R 96 is selected from hydrogen, lower linear alkanyl, lower branched alkanyl, lower cycloalkanyl and lower cycloalkanylmethyl.

94 . The method of claim 1 in which the cancer cell is a tumor cell.

95 . The method of claim 94 in which the tumor cell is a bladder, lung, colon, breast, prostate, pancreatic, ovarian or hepatic tumor cell.

96 . The method of claim 1 which is practiced in vivo as a therapeutic approach towards the treatment of cancer.

97 . The method of claim 96 in which the cancer is a metastatic tumor.

98 . The method of claim 97 in which the cancer is selected from the group consisting of breast, colon, pancreatic, lung, neural, esophageal, gastric, and melanoma.

99 . The method of claim 96 in which the compound is administered in the form of a pharmaceutical composition.

100 . The method of claim 99 in which the compound is administered orally or intravenously.

101 . The method of claim 100 in which the subject is a human.