Diaryl ureas with kinase inhibiting activity

1 . A method of assessing the efficacy of a compound of formula I in treating a treating a disease in a mammalian subject, or a cell derived therefrom, comprising:

measuring the expression or activity of Raf, VEGFR-2, VEGFR-3, p38, PDGFR-beta, and/or Flt-3 in a sample obtained from said subject who has been treated with a compound of formula I, and

determining the effects of said compound on said expression or activity,

wherein said compound of formula I is:

B—NH—C(O)—NH-L-M-L 1 -(Q) 1-3   (I)

wherein B is

(i) phenyl, optionally substituted with 1-3 substituents independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 , C(O)R 1 , C(O)OR 1 , C(O)NR 1 R 2 , NR 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano, and nitro;

(ii) naphthyl, optionally substituted with 1-3 substituents independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 , C(O)R 1 , C(O)OR 1 , C(O)NR 1 R 2 , NR 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano, and nitro;

(iii) a 5 or 6 membered monocyclic heteroaryl group, having 1-3 heteroatoms independently selected from the group consisting of O, N and S, optionally substituted with 1-3 substituents independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 , C(O)R 1 , C(O)OR 1 , C(O)NR 1 R 2 , NR 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano, oxo, and nitro; or

(iv) an 8 to 10 membered bicyclic heteroaryl group in which the first ring is bonded to the NH of FIG. 1 and contains 1-3 heteroatoms independently selected from the group consisting of O, N, and S, and the second ring is fused to the first ring using 3 to 4 carbon atoms. The bicyclic heteroaryl group is optionally substituted with 1-3 substituents independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 , C(O)R 1 , C(O)OR 1 , C(O)NR 1 R 2 , NR 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano, oxo, and nitro.

L is

(i) phenyl, optionally substituted with 1-3 substituents independently selected from the group consisting of C 1 -C 5 linear or branched alkyl, C 1 -C 5 linear or branched haloalkyl, C 1 -C 3 alkoxy, hydroxy, amino, C 1 -C 3 alkylamino, C 1 -C 6 dialkylamino, halogen, cyano, and nitro;

(ii) naphthyl, optionally substituted with 1-3 substituents independently selected from the group consisting of C 1 -C 5 linear or branched alkyl, C 1 -C 5 linear or branched haloalkyl, C 1 -C 3 alkoxy, hydroxy, amino, C 1 -C 3 alkylamino, C 1 -C 6 dialkylamino, halogen, cyano, and nitro;

(iii) a 5 or 6 membered monocyclic heteroaryl group, having 1-3 heteroatoms independently selected from the group consisting of O, N and S, optionally substituted with 1-3 substituents independently selected from the group consisting of C 1 -C 5 linear or branched alkyl, C 1 -C 5 linear or branched haloalkyl, C 1 -C 3 alkoxy, hydroxy, amino, C 1 -C 3 alkylamino, C 1 -C 6 dialkylamino, halogen, cyano, and nitro; or

(iv) an 8 to 10 membered bicyclic heteroaryl group having 1-6 heteroatoms independently selected from the group consisting of O, N and S, optionally substituted with 1-3 substituents independently selected from the group consisting of C 1 -C 5 linear or branched alkyl, C 1 -C 5 linear or branched haloalkyl, C 1 -C 3 alkoxy, hydroxy, amino, C 1 -C 3 alkylamino, C 1 -C 6 dialkylamino, halogen, cyano, and nitro.

M is

(a) —(CH 2 ) m —O—(CH 2 ) l —,

(b) —(CH 2 ) m —(CH 2 ) l —,

(c) —(CH 2 ) m —C(O)—(CH 2 ) l —,

(d) —(CH 2 ) m —NR 3 —(CH 2 ) l —,

(e) —(CH 2 ) m —NR 3 C(O)—(CH 2 ) l —,

(f) —(CH 2 ) m —S—(CH 2 ) l —,

(g) —(CH 2 ) m —C(O)NR 3 —(CH 2 ) l —,

(h) —(CH 2 ) m —CF 2 —(CH 2 ) l —,

(i) —(CH 2 ) m —CCl 2 —(CH 2 ) l —,

(j) —(CH 2 ) m —CHF—(CH 2 ) l —,

(k) —(CH 2 ) m —CH(OH)—(CH 2 ) l —;

(l) —(CH 2 ) m —C≡C—(CH 2 ) l —;

(m) —(CH 2 ) m —C═C—(CH 2 ) l —;

(n) —(CH 2 ) m —CR 4 R 5 —(CH 2 ) l —;

or

(o) a single bond, where m and l are 0;

wherein the variables m and l are integers independently selected from 0-4,

L′ is

(i) phenyl, optionally substituted with 1-2 additional substituents other than Q, independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 , NR 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano and nitro;

(ii) naphthyl, optionally substituted with 1-2 additional substituents other than Q, independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 , NR 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano and nitro;

(iii) a 5 and 6 membered monocyclic heteroaryl group, having 1-3 heteroatoms independently selected from the group consisting of O, N and S, optionally substituted with 1-2 additional substituents other than Q, independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 , NR 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano and nitro and also oxides (e.g. ═O, —O − or —OH);

(iv) an 8 to 10 membered bicyclic heteroaryl group, having 1-6 heteroatoms independently selected from the group consisting of O, N and S, optionally substituted with 1-2 additional substituents other than Q, independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 , NR 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano and nitro and also oxides (e.g. ═O, —O − or —OH).

(v) a saturated and partially saturated C 3 -C 6 monocyclic carbocyclic moiety optionally substituted with 1-2 additional substituents other than Q, independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 , NR 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano and, nitro;

(vi) a saturated and partially saturated C 8 -C 10 bicyclic carbocyclic moiety, optionally substituted with 1-2 additional substituents other than Q, independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 , NR 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano and nitro;

(vii) a saturated and partially saturated 5 and 6 membered monocyclic heterocyclic moiety, having 1-3 heteroatoms independently selected from the group consisting of O, N and S, optionally substituted with 1-2 additional substituents other than Q, independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 N, R 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano and nitro, and also oxides (e.g. ═O, —O − or —OH); or

(viii) a saturated and partially saturated 8 to 10 membered bicyclic heterocyclic moiety, having 1-6 heteroatoms independently selected from the group consisting of O, N and S, optionally substituted with 1-2 additional substituents other than Q, independently selected from the group consisting of R 1 , OR 1 , NR 1 R 2 , S(O) q R 1 , SO 2 NR 1 R 2 , NR 1 SO 2 R 2 , NR 1 C(O)R 2 , NR 1 C(O)OR 2 , halogen, cyano and nitro, and also oxides (e.g. ═O, —O − or —OH); and

each Q is independently C(O)R 4 , C(O)OR 4 and C(O)NR 4 R 5 ;

wherein each R 1 -R 5 is independently selected from:

(a) hydrogen,

(b) C 1 -C 5 linear, branched, or cyclic alkyl,

(c) phenyl,

(d) C 1 -C 3 alkyl-phenyl, wherein the alkyl moiety is optionally substituted with halogen up to per-halo;

(e) up to per-halo substituted C 1 -C 5 linear or branched alkyl. Or

(f) —(CH 2 ) q —X, where X is a 5 or 6 membered monocyclic heterocyclic ring, containing 1-4 atoms selected from oxygen, nitrogen and sulfur, which is saturated, partially saturated, or aromatic, or a 8-10 membered bicyclic heteroaryl having 1-4 heteroatoms selected from the group consisting of O, N and S; and wherein said alkyl moiety is optionally substituted with halogen up to per-halo,

wherein each R 1 -R 5 , other than per-halo substituted C 1 -C 5 linear or branched alkyl, is optionally substituted with 1-3 substituents independently selected from the group consisting of C 1 -C 5 linear or branched alkyl, up to perhalo substituted C 1 -C 5 linear or branched alkyl, C 1 -C 3 alkoxy, hydroxy, carboxy, amino, C 1 -C 3 alkylamino, C 1 -C 6 dialkylamino, halogen, cyano, and nitro;

wherein the variable p is an integer selected from 0, 1, or 2 and the variable q is an integer selected from 0, 1, 2, 3, or 4.

2 . A method of claim 1 , wherein said measuring the expression is determining the amounts of mRNA corresponding to Raf, VEGFR-2, VEGFR-3, p38, PDGFR-beta, and/or Flt-3.

3 . A method of claim 1 , wherein said measuring the expression is determining the amounts of polypeptide corresponding to Raf, VEGFR-2, VEGFR-3, p38, PDGFR-beta, and/or Flt-3.

4 . A method of claim 3 , wherein said measuring the activity is determining the amounts of phospho-ERK.

5 . A method of claim 4 , wherein said subject has cancer, and measuring the activity is determining the amounts of phospho-ERK in peripheral blood lymphocytes or a tissue biopsy of a cancer.

6 . A method of claim 1 , further comprising comparing the expression or activity in said sample to a normal control.

7 . A method of claim 1 , further comprising comparing the expression or activity in at least one sample before treating with said compound and in at least one sample after treating with said compound.

8 . A method of claim 1 , further comprising measuring expression in at least two different samples collected at different timepoints in the treatment regimen with said compound.

9 . A method of claim 1 , wherein a reduction in expression or activity indicates that said compound is effective in treating said disease.

10 . A method of claim 1 , wherein said disease is renal cell carcinoma or melanoma.

11 . A method of claim 1 , wherein said sample comprises tumor cells.

12 . A method of claim 1 , wherein said sample comprises peripheral blood cells.

13 . A method of claim 1 , further comprising administering a compound of formula I at a plurality of timepoints.

14 . A method of selecting subjects having a disease for treatment with a compound of formula I, comprising:

measuring the expression or activity of Raf, VEGFR-2, VEGFR-3, p38, PDGFR-beta, and/or Flt-3, in a sample obtained from a subject having a disease, and administering said compound of formula I to subjects who are identified as having high levels of expression or activity, where said compound is a compound of formula I of claim 1 .

15 . A method of selecting subjects having a disease for treatment with a compound of formula I, comprising:

determining the presence of a Raf, VEGFR-2, VEGFR-3, p38, PDGFR-beta, and/or Flt-3 gene mutation in a sample obtained from a subject, wherein said mutation is associated with a disease, and

administering said compound of formula I of claim 1 to subjects who are identified as having said mutation.

16 . A method of claim 15 , wherein said mutation is in the BRAF gene.

17 . A method of claim 16 , wherein said BRAF mutation is at amino acid position 599 of the coding sequence of said gene.

18 . A method of claim 17 , wherein said BRAF mutation is V599E.

19 . A method of claim 15 , wherein said disease is melanoma.

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24 . A method of inhibiting lymphangiogenesis, comprising:

administering administering a compound of formula I of claim 1 to a subject in need thereof.

25 . A method of inhibiting angiogenesis, comprising:

administering administering a compound of formula I of claim 1 to a subject in need thereof.

26 . (canceled)

27 . A method treating a tumor in a subject in need thereof, comprising:

administering an effective amount of a compound of formula I of claim 1 to a subject in need thereof, wherein said amount is effective to inhibit tumor cell proliferation and neovascularization.

28 . (canceled)

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31 . A method of assessing the efficacy of a compound in treating a disease in a mammalian subject, or a cell derived therefrom, comprising:

measuring the expression or activity of Raf, VEGFR-2, VEGFR-3, p38, PDGFR-beta, and/or Flt-3 in a sample obtained from said subject who has been treated with said compound, and

determining the effects of said compound on said expression or activity,

wherein said compound is:

N-(4-chloro-3-(trifluoromethyl)phenyl)-N′-(4-(2-(N-methylcarbamoyl)-4-pyridyloxy)phenyl)urea,

N-(4-bromo-3-(trifluoromethyl)phenyl)-N′-(4-(2-(N-methylcarbamoyl)-4-pyridyloxy)phenyl)urea,

N-(4-bromo-3-(trifluoromethyl)phenyl)-N′-(4-(2-(N-methylcarbamoyl)-4-pyridyloxy)-2-chlorophenyl)urea,

N-(4-chloro-3-(trifluoromethyl)phenyl)-N′-(4-(2-carbamoyl-4-pyridyloxy)phenyl)urea,

N-(4-chloro-3-(trifluoromethyl)phenyl)N′-(4-(1-hydroxy-2-carbamoyl-4-pyridyloxy)phenyl)urea,

N-(4-chloro-3-(trifluoromethyl)phenyl)N′-(4-(1-hydroxy-2-(N-methylcarbamoyl)-4-pyridyl oxy)phenyl)urea,

N-(4-chloro-3-(trifluoromethyl)phenyl)N′-(4-(2-(N-methylcarbamoyl)-4-pyridyl oxy)-2-fluorophenyl)urea,

N-(4-bromo-3-(trifluoromethyl)phenyl)-N′-(4-(2-(N-methylcarbamoyl)-4-pyridyl oxy)-2-fluorophenyl)urea,

N-(4-fluoro-3-(trifluoromethyl)phenyl)-N′-(4-(2-(N-methylcarbamoyl)-4-pyridyl oxy)-2-fluorophenyl)urea,

N-(4-chloro-3-(trifluoromethyl)phenyl)-N′-(4-(2-(N-methylcarbamoyl)-4-pyridyl oxy)-2-chlorophenyl)urea,

N-(6-(2,2,4,4-tetrafluoro-4H-benzo[1,3]dioxinyl))N′-(4-(2-cyano-4-pyridyloxy)phenyl)urea, or

N-(6-(2,2,4,4-tetrafluoro-4H-benzo[1,3]dioxinyl))N′-(4-(2-cyano-4-pyridyloxy)-2-fluorophenyl)urea.

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48 . A method of treating acute myeloid leukemia, comprising

administering an effective amount of a compound of formula I of claim 1 .

49 . A method of treating a hematopoietic cell cancer, comprising

administering an effective amount of a compound of formula I of claim 1 .

50 . A method of claim 49 , wherein said cancer is associated with a mutation in Flt-3.

51 . A method of claim 50 , wherein said mutation is FLT3 ITD.

52 . A method of claim 15 , wherein said disease is a cancer and the presence of a Flt-3 mutation is determined.