HETEROCYCLIC COMPOUNDS AS KINASE INHIBITORS FOR THERAPEUTIC USES

Abstract

Heterocyclic compounds of formula I shown below and pharmaceutical compositions containing one of such compounds: Also disclosed is a method of treating a condition modulated by the colony-stimulating factor-1 receptor with one of the heterocyclic compounds.

##STR00001##

Claims

1. A compound of formula I: ##STR00135## wherein A is H, C.sub.1-C.sub.6 alkyl, or ORr, Rr being H or C.sub.1-6 alkyl; Y.sup.1 is phenyl substituted with (R.sup.1).sub.n, 5-membered heteroaryl substituted with (R.sup.2).sub.o, 5-membered heterocycloalkenyl substituted with (R.sup.2).sub.o, or alkenyl substituted with (R.sup.2).sub.o, in which R.sup.1 in (R.sup.1).sub.n, n being 0-4, is, independently, F, Cl, Br, NO.sub.2, CN, amino, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.5-C.sub.15 heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl, or spiroamino; and R.sup.2 in (R.sup.2).sub.o, o being 0-5, is, independently, F, Cl, Br, NO.sub.2, CN, amino, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.5-C.sub.15 heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl, spiroamino, or C.sub.1-C.sub.6 alkoxyl; X.sup.1 is N or CR.sup.3, R.sup.3 being H, F, Cl, Br, CN, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, or C.sub.1-C.sub.6 alkoxyl; X.sup.2 is O, S, NH, or CH.sub.2; Y.sup.2is ##STR00136## in which each of Q.sup.1, Q.sup.2, Q.sup.3, Q.sup.4, Q.sup.5, Q.sup.6, Q.sup.7, and Q.sup.8 is, independently, N or CR.sup.4, R.sup.4 being H, F, Cl, Br, CN, amino, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, or C.sub.1-C.sub.6 alkoxyl; Z.sup.1 is O, S, or NRr; Z.sup.2 is O, S, or NRr; and G and H are, respectively, C or N and N or C; X.sup.3 is deleted, CH.sub.2, (CH.sub.2).sub.2, or CH(C≡CH); Y.sup.3 is C.sub.1-C.sub.6 alkyl, aryl, heteroaryl, C.sub.3-C.sub.8 cycloalkyl, or C.sub.5-C.sub.6 heterocycloalkyl having one heteroatom, in which the one heteroatom is O or N; and X.sup.4 in (X.sup.4).sub.m, m being 0-5, is, independently, F, Cl, Br, CN, SO.sub.2NH.sub.2, amino, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, or C.sub.1-C.sub.6 alkoxyl.

2. The compound of claim 1, wherein Y.sup.1 is phenyl substituted with (R.sup.1).sub.n, 5-membered heteroaryl substituted with (R.sup.2).sub.o, or 5-membered heterocycloalkenyl substituted with (R.sup.2).sub.o, in which R.sup.1 in (R.sup.1).sub.n is, independently, F, Cl, Br, NO.sub.2, CN, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.5-C.sub.15 heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl, or spiroamino; and R.sup.2 in (R.sup.2).sub.o is, independently, F, Cl, Br, NO.sub.2, CN, amino, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.5-C.sub.15 heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl, spiroamino, or C.sub.1-C.sub.6 alkoxyl.

3. The compound of claim 2, wherein Y.sup.2 is ##STR00137## Y.sup.3 is pyridyl, and R.sup.1 is C.sub.5-C.sub.15 heterocycloalkyl.

4. The compound of claim 1, wherein Y.sup.2 is ##STR00138## in which Z.sup.2 is O or NRr.

5. The compound of claim 4, wherein Y.sup.2 is ##STR00139## Y.sup.3 is pyridyl, and R.sup.1 is amino.

6. The compound of claim 1, wherein the compound is of formula Ia: ##STR00140## in which R.sup.1 is amino or C.sub.5-C.sub.15 heterocycloalkyl.

7. The compound of claim 6, wherein Y.sup.2 is ##STR00141## Y.sup.3 is pyridyl; X.sup.3 is CH.sub.2; X.sup.4 is CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, or OCH.sub.3; and m is 1.

8. The compound of claim 6, wherein Y.sup.2 is ##STR00142## Y.sup.3 is phenyl; X.sup.3 is CH.sub.2; each of X.sup.4 is, independently, F, Cl, Br, CN, SO.sub.2NH.sub.2, CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, OCF.sub.3, C.sub.1-C.sub.6 alkoxyl, or amino; and m is 0-2.

9. The compound of claim 6, wherein Y.sup.2 is ##STR00143## Y.sup.3 is phenyl; X.sup.3 is deleted; each of X.sup.4 is, independently, F, Cl, Br, CN, SO.sub.2NH.sub.2, CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, OCF.sub.3, C.sub.1-C.sub.6 alkoxyl, or amino; and m is 0-2.

10. The compound of claim 7, wherein R.sup.1 is amino.

11. The compound of claim 6, wherein Y.sup.2 is ##STR00144## Y.sup.3 is phenyl or pyridyl; and X.sup.3 is CH.sub.2.

12. The compound of claim 11, wherein Y.sup.2 is ##STR00145##

13. The compound of claim 6, wherein Y.sup.2 is ##STR00146## Y.sup.3 is phenyl or pyridyl; and X.sup.3 is CH.sub.2.

14. The compound of claim 13, wherein Y.sup.2 is ##STR00147##

15. The compound of claim 1, wherein the compound is of formula Ib: ##STR00148##

16. The compound of claim 15, wherein Y.sup.1 is ##STR00149## Y.sup.3 is phenyl or pyridyl; and X.sup.3 is deleted or CH.sub.2.

17. The compound of claim 16, wherein each of X.sup.4 is, independently, CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, or OCH3 and m is 0-2.

18. The compound of claim 15, wherein Y.sup.1 is ##STR00150## Y.sup.3 is phenyl or pyridyl; and X.sup.3 is deleted or CH.sub.2, in which Q.sup.9 is N or CR.sup.5, R.sup.5 being H, F, Cl, Br, CN, amino, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, or C.sub.1-C.sub.6 alkoxyl.

19. The compound of claim 18, wherein each of X.sup.4 is, independently, CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, or OCH3 and m is 0-2.

20. The compound of claim 15, wherein Y.sup.1 is ##STR00151## Y.sup.3 is phenyl or pyridyl; and X.sup.3 is deleted or CH.sub.2.

21. The compound of claim 20, wherein each of X.sup.4 is, independently, CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, or OCH.sub.3 and m is 0-2.

22. The compound of claim 1, wherein the compound is one of the following compounds: ##STR00152## ##STR00153##

23. The compound of claim 1, wherein the compound is ##STR00154## ##STR00155##

24. The compound of claim 1, wherein the compound is ##STR00156##

25-31. (canceled)

Description

DETAILED DESCRIPTION

[0021] Disclosed first in detail are heterocyclic compounds of formula I:

##STR00004##

[0022] A, Y.sup.1, X.sup.1, X.sup.2, Y.sup.2, X.sup.3, Y.sup.3, X.sup.4, and m are defined in the SUMMARY section above.

[0023] In one embodiment, the compounds of formula I have Y.sup.1 being phenyl substituted with (R.sup.1).sub.n, 5-membered heteroaryl substituted with (R.sup.2).sub.o, or 5-membered heterocycloalkenyl substituted with (R.sup.2).sub.o, in which R.sup.1 in (R.sup.1).sub.n is, independently, F, Cl, Br, NO.sub.2, CN, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.5-C.sub.15 heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl, or spiroamino; and R.sup.2 in (R.sup.2).sub.o is, independently, F, Cl, Br, NO.sub.2, CN, amino, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.5-C.sub.15 heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl, spiroamino, or C.sub.1-C.sub.6 alkoxyl.

[0024] In one subset of this embodiment, the compounds of formula I have Y.sup.2 being

##STR00005##

Y.sup.3 being pyridyl, and R.sup.1 being C.sub.5-C.sub.15 heterocycloalkyl.

[0025] In another embodiment, the compounds of formula I have Y.sup.2 being

##STR00006##

in which Z.sup.2 is O or NRr.

[0026] In one subset of this embodiment, the compounds of formula I have Y.sup.2 being

##STR00007##

Y.sup.3 being pyridyl, and R.sup.1 being amino.

[0027] In still another embodiment, the compounds of formula I are those covered by formula Ia:

##STR00008##

in which R.sup.1 is amino or C.sub.5-C.sub.15 heterocycloalkyl.

[0028] In one subset of this embodiment, the compounds of formula Ia have Y.sup.2 being

##STR00009##

Y.sup.3 being pyridyl; X.sup.3 being CH.sub.2; X.sup.4 being CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, or OCH.sub.3, m being 1; and R.sup.1 preferably being amino.

[0029] In another subset, the compounds have Y.sup.2 being

##STR00010##

Y.sup.3 being phenyl; X.sup.3 being CH.sub.2; each of X.sup.4 being, independently, F, Cl, Br, CN, SO.sub.2NH.sub.2, CH.sub.3, CH2F, CHF2, CF.sub.3, OCF.sub.3, C.sub.1-C.sub.6 alkoxyl, or amino; and m being 0-2.

[0030] In a third subset, the compounds have Y.sup.2 being

##STR00011##

Y.sup.3 being phenyl; X.sup.3 being deleted; each of X.sup.4 being, independently, F, Cl, Br, CN, SO.sub.2NH.sub.2, CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, OCF.sub.3, C.sub.1-C.sub.6 alkoxyl, or amino; and m being 0-2.

[0031] Formula Ia includes compounds in which Y.sup.2 is

##STR00012##

Y.sup.3 is phenyl or pyridyl, and X.sup.3 is CH.sub.2. As an example, Y.sup.2 is

##STR00013##

[0032] Formula Ia further includes compounds in which Y.sup.2 is

##STR00014##

Y.sup.3 is phenyl or pyridyl, and X.sup.3 is CH.sub.2. For instance, Y.sup.2 is

##STR00015##

[0033] In a fourth embodiment, the heterocyclic compounds of this invention are covered by formula Ib:

##STR00016##

[0034] A subset of compounds of formula Ib have Y.sup.1 being

##STR00017##

Y.sup.3 being phenyl or pyridyl; and X.sup.3 being deleted or CH.sub.2.

[0035] In a different subset, compounds of formula Ib have Y.sup.1 being

##STR00018##

Y.sup.3 being phenyl or pyridyl; and X.sup.3 being deleted or CH.sub.2, in which Q.sup.9 is N or CR.sup.5, R.sup.5 being H, F, Cl, Br, CN, amino, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, or C.sub.1-C.sub.6 alkoxyl.

[0036] In another subset, compounds of this formula have Y.sup.1 being

##STR00019##

Y.sup.3 being phenyl or pyridyl; and X.sup.3 being deleted or CH.sub.2.

[0037] Typically, the compounds of formula Ib have each of X.sup.4 being, independently, CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, or OCH.sub.3 and m being 0-2.

[0038] Exemplary compounds of formula I include, but are not limited to, the following compounds:

##STR00020## ##STR00021## ##STR00022## ##STR00023##

[0039] The compounds of formula I can be prepared according to methods well known in the field. See, for example, R. Larock, Comprehensive Organic Transformations (2nd Ed., VCH Publishers 1999); P. G. M. Wuts and T. W. Greene, Greene's Protective Groups in Organic Synthesis (4th Ed., John Wiley and Sons 2007); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis (John Wiley and Sons 1994); L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis (2nd ed., John Wiley and Sons 2009); and G. J. Yu et al., J. Med. Chem. 2008, 51, 6044-6054.

[0040] Also within the scope of this invention is a pharmaceutical composition containing one or more of the heterocyclic compounds of formula I. The pharmaceutical composition is used for treating a CSF1R modulated condition.

[0041] In certain embodiments, the pharmaceutical composition further contains one of the following therapeutic agents: an anti-proliferative agent, an anti-inflammatory agent, an immunomodulatory agent, and an immunosuppressive agent.

[0042] In other embodiments, the pharmaceutical composition further contains one of the following therapeutic agents: an alkylating agent, e.g., adozelesin, altretamine, bizelesin, busulfan, carboplatin, carboquone, carmustine, chlorambucil, cisplatin, cyclophosphamide, dacarbazine, estramustine, fotemustine, hepsulfam, ifosfamide, improsulfan, irofulven, lomustine, mechlorethamine, melphalan, oxaliplatin, piposulfan, semustine, streptozocin, temozolomide, thiotepa, and treosulfan; an antibody, e.g., alemtuzumab, bevacizumab, cetuximab, galiximab, gemtuzumab, nivolumab, panitumumab, pembrolizumab, pertuzumab, rituximab, tositumomab, trastuzumab, and 90 Y ibritumomab tiuxetan; a targeted signal transduction inhibitor, e.g., bortezomib, geldanamycin, and rapamycin; a kinase inhibitor, e.g., erlotinib, gefitinib, flavopiridol, imatinib mesylate, lapatinib, sorafenib, sunitinib malate, AEE-788, AG-013736, AMG 706, AMN107, BMS-354825, BMS-599626, 7-hydroxystaurosporine, vemurafenib, dabrafenib, trametinib, cobimetinib, selumetinib, and vatalanib; a taxane, e.g., DJ-927, docetaxel, TPI 287, paclitaxel, and DHA-paclitaxel; a retinoid, e.g., alitretinoin, bexarotene, fenretinide, isotretinoin, and tretinoin; an alkaloid, e.g., etoposide, homoharringtonine, teniposide, vinblastine, vincristine, vindesine, and vinorelbine; an antibiotic, e.g., bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, menogaril, mitomycin, mitoxantrone, neocarzinostatin, pentostatin, and plicamycin; an antiangiogenic agent, e.g., AE-941, ABT-510, 2-methoxyestradiol, lenalidomide, and thalidomide; a topoisomerase inhibitor, e.g., amsacrine, edotecarin, exatecan, irinotecan, 7-ethyl-10-hydroxy-camptothecin, rubitecan, topotecan, and 9-aminocamptothecin; an antimetabolite, e.g., azacitidine, capecitabine, cladribine, clofarabine, cytarabine, decitabine, floxuridine, fludarabine, 5-fluorouracil, ftorafur, gemcitabine, hydroxyurea, mercaptopurine, methotrexate, nelarabine, pemetrexed, raltitrexed, thioguanine, and trimetrexate; a hormone or hormone antagonist, e.g., anastrozole, androgens, buserelin, diethylstilbestrol, exemestane, flutamide, fulvestrant, goserelin, idoxifene, letrozole, leuprolide, magestrol, raloxifene, tamoxifen, and toremifene; a biological response modifier, e.g., imiquimod, interferon-α, and interleukin-2; an indoleamine 2,3-dioxygenase inhibitor; a chemotherapeutic agent, e.g., 3-amino-2-carboxy aldehyde thiosemicarbazone, altrasentan, aminoglutethimide, anagrelide, asparaginase, bryostatin-1, cilengitide, elesclomol, eribulin mesylate, ixabepilone, lonidamine, masoprocol, mitoguanazone, oblimersen, sulindac, testolactone, and tiazofurin; a mammalian target of rapamycin inhibitor; a phosphoinositide 3-kinase inhibitor; a cyclin-dependent kinase 4 inhibitor; a protein kinase B inhibitor; a heat shock protein 90 inhibitor; a farnesyltransferase inhibitor; an aromatase inhibitor (such as anastrozole, letrozole, and exemestane); a mitogen-activated protein kinase kinase inhibitor; a tyrosine kinase inhibitor; an epidermal growth factor receptor inhibitor; a programmed cell death protein 1 inhibitor; a programmed death-ligand 1 inhibitor; or an interleukin 8 receptor beta inhibitor.

[0043] Still within the scope of this invention is a method of treating a CSF1R modulated condition using one or more of the above-described heterocyclic compounds. For example, the condition can be a cancer, e.g., acute myeloid leukemia, bladder cancer, breast cancer, cervical cancer, colon cancer, gastric cancer, gastrointestinal stromal tumor, glioblastoma multiforme, hepatocellular carcinoma, Hodgkin's lymphoma, kidney cancer, liver cancer, lung cancer, melanoma, metastatic tumor, ovarian cancer, pancreatic cancer, pigmented villondular synovitis, prostate cancer, tenosynovial giant cell tumors, endometrial cancer, multiple myeloma, myelocytic leukemia, bone cancer, renal cancer, brain cancer, myeloproliferative disorder, esophageal cancer, squamous cell carcinoma, uveal melanoma, follicular lymphoma, colorectal cancer, head and neck cancer, astrocytoma, and pulmonary adenocarcinoma; an inflammatory disorder or an autoimmune disease, e.g., psoriatic arthritis, arthritis, asthma, thyroiditis, glomerular nephritis, atherosclerosis, psoriasis, Sjogren's syndrome, rheumatoid arthritis, systemic lupus erythematosis, cutaneous lupus erythematosus, Crohn's disease, ulcerative colitis, type I diabetes, multiple sclerosis, human immunodeficiency virus encephalitis, Alzheimer's disease, amyotrophic lateral sclerosis, and epilepsy; or a bone disorder, e.g., osteoporosis, osteoarthritis, periodontitis, periprosthetic osteolysis, and Paget's disease.

[0044] Without further elaboration, it is believed that one skilled in the art can, based on the above description, utilize the present invention to its fullest extent. The following specific examples are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever. All publications cited herein are incorporated by reference in their entirety.

EXAMPLE 1: SYNTHESIS OF HETEROCYCLIC COMPOUNDS

[0045] Exemplary compounds of this invention, shown in Table 1 below, were prepared by procedures shown in Scheme 1, Scheme 2, Scheme 3, or Scheme 4. Table 1 includes mass spectral data for the compounds.

[0046] All chemicals and solvents were purchased from commercial suppliers and used as received. All reactions were carried out under an atmosphere of dry nitrogen unless specified otherwise. Reactions were monitored by thin layer chromatography using Merck 60 F254 silica gel glass backed plates (5×10 cm) and zones were detected visually under ultraviolet irradiation (254 nm) or by spraying with phosphomolybdic acid reagent (Aldrich) followed by heating at 80° C. Microwave reactions were performed in the CEM Discover SP System.

[0047] Flash column chromatography was performed by using Merck Kieselgel 60, No. 9385, 230-400 mesh ASTM silica gel as the stationary phase. Proton nuclear magnetic resonance (.sup.1H NMR) spectra were measured on a Varian Mercury-300 or Varian Mercury-400 spectrometer. Chemical shifts were recorded in parts per million (ppm) on the delta (6) scale relative to the resonance of the solvent peak. The following abbreviations were used to describe coupling: s=singlet; d=doublet; t=triplet; q=quartet; quin=quintet; ABq=AB quartet; AA′XX′=second order AA′XX′ pattern; app.=apparent; br=broad; and m=multiplet.

[0048] Liquid chromatography mass spectrometry (LCMS) data was obtained with an Agilent MSD-1100 ESI-MS/MS, an Agilent 1200 series LC/MSD VL, or a Waters Acquity UPLC-ESI-MS/MS system.

##STR00024##

[0049] Among the listed reagents and solvents in Scheme 1, SOCl.sub.2 is thionylchloride, DMF is dimethylformamide, PhMe is toluene, tBuOK is potassium tert-butoxide, THF is tetrahydrofuran, DMSO is dimethylsulfoxide, and CH.sub.2Cl.sub.2 is methylene chloride.

[0050] 4-Chloro-7-fluoroquinazoline (B). To a suspension of 7-fluoroquinazolin-4-ol compound A (6.32 g, 38.5 mmol) in dry PhMe (30 mL), SOCl.sub.2 (22 mL, 7.7 eq.) and DMF (2.6 mL) were added. The resulting mixture was refluxed for 10 h. The mixture was then cooled to room temperature, quenched with water (200 mL), and extracted with ethyl acetate (EtOAc; 170 mL). The combined organic extracts were washed with water (300 mL) and brine (30 mL), dried over sodium sulfate (NA.sub.2SO.sub.4), and concentrated to afford compound B (6.08 g, 86%) as a yellow solid. LCMS (ESI) m/z calculated for C.sub.8H.sub.4ClFN.sub.2: 182, 184; found: 183, 185 [M+H].sup.+. .sup.1H-NMR (400 MHz, CDCl.sub.3) 6 9.04 (s, 1H), 8.33 (dd, J=9.2 Hz, .sup.4J.sub.F,H=6.0 Hz, 1H), 7.71 (dd, .sup.3J.sub.F,H=9.2 Hz, J=2.4 Hz, 1H), 7.52 (ddd, J=9.2, 2.4 Hz, .sup.3J.sub.F,H=8.4 Hz, 1H).

[0051] 4-((7-fluoroquinazolin-4-yl)oxy)aniline (C). A mixture of 4-aminophenol F (3.05 g, 28.0 mmol) and t-BuOK (3.14 g, 28.0 mmol) in dry THF (100 mL) was stirred for 20 min at 0° C. Subsequently, compound B (4.44 g, 24.3 mmol) was added slowly in small portions. The reaction mixture was then stirred for 3 h at 0° C., during which a suspension was formed. Afterwards, the suspension was filtered through a pad of celite, The pad was rinsed with THF. THF in the filtrate was removed by evaporation to give a crude residue. The residue was suspended in methanol (MeOH) and sonicated to form a solid. The solid was collected via filtration and dried to afford the titled product C (5.47 g, 88%) as an off-white solid. LCMS (ESI) m/z calculated for C.sub.14H.sub.10BrN.sub.3O: 255; found: 256 [M+H].sup.+. .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.76 (s, 1H), 8.40 (dd, J=9.2 Hz, .sup.4J.sub.F,H=6.0 Hz, 1H), 7.61 (dd, .sup.3J.sub.F,H=9.2 Hz, J=2.4 Hz, 1H), 7.40 (ddd, J=9.2, 2.4 Hz, .sup.3J.sub.F,H=8.8 Hz, 1H), 7.04 (AA′XX′, J.sub.AX=8.8 Hz, J.sub.AX′=0 Hz, 2H), 6.77 (AA′XX′, J.sub.AX=8.8 Hz, J.sub.AX′=0 Hz, 2H), 3.71 (br, 2H).

[0052] 4-(4-aminophenoxy)-N,N-dimethylquinazolin-7-amine (D). To a solution of compound C (600 mg, 2.35 mmol) in DMSO (7.1 mL), 2 M dimethlyamine in THF (3 eq., 3.53 mL) was added. The mixture was then irradiated in microwave for 10 min at 170° C. After compound C was consumed, the reaction mixture was diluted with EtOAc (30 mL) and 2% sodium carbonate (NA.sub.2CO.sub.3) solution (80 mL), and extracted with EtOAc (50 mL). The combined organic extracts were sequentially washed with 2% NA.sub.2CO.sub.3 solution (160 mL), water (200 mL), and brine (20 mL), dried over NA.sub.2SO.sub.4, and concentrated. The crude residue was suspended in MeOH. Particles in the supension were collected via filtration and dried to afford the titled product D (207 mg, 32%) as a pale yellow solid. LCMS (ESI) m/z calculated for C.sub.16H.sub.16N.sub.4O: 280; found: 281 [M+H].sup.+. .sup.1H-NMR (300 MHz, CDCl.sub.3): δ 8.58 (s, 1H), 8.15 (d, J=9.2 Hz, 1H), 7.12 (dd, J=9.2 Hz, J=2.7 Hz, 1H), 7.03 (AA′XX′, J.sub.AX=9.0 Hz, J.sub.AX′=0 Hz, J.sub.AA′=2.9 Hz, J.sub.XX′=2.9 Hz, 2H), 6.96 (d, J=2.7 Hz, 1H), 6.75 (AA′XX′, J.sub.AX=9.0 Hz, J.sub.AX′=0 Hz, J.sub.AA′=2.9 Hz, J.sub.XX′=2.9 Hz, 2H), 3.67 (br, 2H), 3.15 (s, 6H).

[0053] 1-(4-((7-(dimethylamino)quinazolin-4-yl)oxy)phenyl)-3-phenylurea (compound 4, E). To a solution of compound D (165 mg, 0.588 mmol) in CH.sub.2Cl.sub.2 (15 mL), compound H (108 μL, 0.883 mmol) was added. The mixture was refluxed for 10 h, during which a suspension was formed. Particles in the suspension were collected via filtration and rinsed with excess CH.sub.2Cl.sub.2 to afford the titled product E (214 mg, 91%) as a white solid. LCMS (ESI) m/z calculated for C.sub.23H.sub.21N.sub.5O.sub.2: 399; found: 400 [M+H].sup.+. .sup.1H-NMR (400 MHz, DMSO-d.sub.6): δ 8.77 (br s, 1H), 8.71 (br s, 1H), 8.45 (s, 1H), 8.11 (d, J=9.6 Hz, 1H), 7.52 (AA′XX′, J.sub.AX=8.8 Hz, J.sub.AX′=0 Hz, J.sub.AA′=2.6 Hz, J.sub.XX′=2.6 Hz, 2H), 7.47 (d, J=5.6 Hz, 2H), 7.32-7.27 (m, 3H), 7.19 (AA′XX′, J.sub.AX=8.8 Hz, J.sub.AX′=0 Hz, J.sub.AA′=2.6 Hz, J.sub.XX′=2.6 Hz, 2H), 6.97 (t, J=7.2 Hz, 1H), 6.88 (d, J=2.4 Hz, 1H), 3.12 (s, 6H).

[0054] Compounds 1, 2, 3, 5, 6, 7, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 52, and 106 were prepared in a manner similar to compound 4, with appropriate amines G and isocyanates H.

##STR00025##

[0055] Among the listed reagents and solvents in Scheme 2, Py is pyridine, and Et.sub.3N is triethylamine.

[0056] 4-nitrophenyl ((6-(trifluoromethyl)pyridin-3-yl)methyl)carbamate (J). To a −30° C. solution of (6-(trifluoromethyl)pyridin-3-yl)methanamine I (2.69 g, 15.3 mmol) and Py (1.23 mL, 15.3 mmol) in CH.sub.2Cl.sub.2 (50 mL), 4-nitrophenyl chloroformate (3.85 g, 19.1 mmol) was slowly added. The reaction mixture was stirred for 8 h and allowed to slowly warm to 0° C. Subsequently, water (50 mL) was added to the reaction mixture. The mixture was then stirred for 20 min, during which a suspension was formed. Particles in the suspension were removed by filtering the reaction mixture through a pad of celite. The filtrate was washed with 2% sodium bisulfate solution (20 mL), 2% sodium bicarbonate solution (40 mL), water (20 mL), and brine (3 mL), dried over NA.sub.2SO.sub.4, and concentrated. The crude residue was purified by column chromatography to afford the titled product J (3.33 g, quant.) as a white solid. LCMS (ESI) m/z calculated for C.sub.14H.sub.10F.sub.3N.sub.3O.sub.4: 341; found: 342 [M+H].sup.+. .sup.1H-NMR (400 MHz, CDCl.sub.3): δ 8.73 (d, J=1.4 Hz, 1H), 8.26 (AA′XX′, J.sub.AX=9.2 Hz, J.sub.AX′=0 Hz, J.sub.AA′=2.6 Hz, J.sub.xx′=2.6 Hz, 2H), 7.91 (dd, J=8.0, 1.4 Hz, 1H), 7.71 (d, J=8.0 Hz, 1H), 7.33 (AA′XX′, J.sub.AX=9.2 Hz, J.sub.AX′=0 Hz, J.sub.AA′=2.6 Hz, J.sub.XX′=2.6 Hz, 2H), 5.68 (br t, 1H), 4.58 (d, J=6.4 Hz, 2H).

[0057] 1-(4-((7-(dimethylamino)quinazolin-4-yl)oxy)phenyl)-3-((6-(trifluoromethyl)pyridin-3-yl)methyl)urea (compound 27, K). To a solution of compound J (2.19 g, 6.43 mmol) and compound D (1.06 g, 3.78 mmol) in CH.sub.2Cl.sub.2 (95 mL), Et.sub.3N (1.05 mL) was added. The reaction mixture was refluxed for 4 d, during which a suspension was formed. Particles in the suspension were collected via filtration and rinsed with excess CH.sub.2Cl.sub.2 to afford the titled product K (1.33 g, 73%) as a white solid. LCMS (ESI) m/z calculated for C.sub.24H.sub.21F.sub.3N.sub.6O.sub.2: 482; found: 483 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.83 (s, 1H), 8.72 (d, J=1.2 Hz, 1H), 8.43 (s, 1H), 8.10 (d, J=9.2 Hz, 1H), 8.00 (dd, J=8.4, 1.2 Hz, 1H), 7.89 (d, J=8.4 Hz, 1H), 7.47 (AA′XX′, J.sub.AX=8.8 Hz, J.sub.AX′=0 Hz, J.sub.AA′=2.6 Hz, J.sub.XX′=2.6 Hz, 2H), 7.29 (d, J=9.2, 2.4 Hz, 1H), 7.13 (AA′XX′, J.sub.AX=8.8 Hz, J.sub.AX′=0 Hz, J.sub.AA′=2.6 Hz, J.sub.XX′=2.6 Hz, 2H), 6.87 (d, J=2.4 Hz, 1H), 6.84 (t, J=6.0 Hz, 1H), 4.44 (d, J=6.0 Hz, 2H), 3.12 (s, 6H).

[0058] 1-(4-((7-(dimethylamino)quinazolin-4-yl)oxy)phenyl)-3-((6-methylpyridin-3-yl)methyl)urea (compound 65, K) LCMS (ESI) m/z calculated for C.sub.24H.sub.21F.sub.3N.sub.6O.sub.2: 428; found: 429 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.68 (s, 1H), 8.43 (s, 1H), 8.39 (d, J=2.4 Hz, 1H), 8.10 (d, J=9.2 Hz, 1H), 7.61 (dd, J=8.0, 2.4 Hz, 1H), 7.46 (AA′XX′, J.sub.AX=8.8 Hz, 2H), 7.29 (dd, J=9.2, 2.4 Hz, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.13 (AA′XX′, J.sub.AX=8.8 Hz, 2H), 6.87 (d, J=2.4 Hz, 1H), 6.67 (t, J=6.0 Hz, 1H), 4.28 (d, J=6.0 Hz, 1H), 3.12 (s, 6H), 2.44 (s, 3H).

[0059] 1-(4-((7-(dimethylamino)quinazolin-4-yl)oxy)phenyl)-3-((6-methylpyridin-3-yl)methyl)urea (compound 66, K) LCMS (ESI) m/z calculated for C.sub.24H.sub.21F.sub.3N.sub.6O.sub.2: 444; found: 445 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.68 (s, 1H), 8.43 (s, 1H), 8.39 (d, J=2.4 Hz, 1H), 8.10 (d, J=9.2 Hz, 1H), 7.61 (dd, J=8.0, 2.4 Hz, 1H), 7.46 (AA′XX′, J.sub.AX=8.8 Hz, 2H), 7.29 (dd, J=9.2, 2.4 Hz, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.13 (AA′XX′, J.sub.AX=8.8 Hz, 2H), 6.87 (d, J=2.4 Hz, 1H), 6.67 (t, J=6.0 Hz, 1H), 4.28 (d, J=6.0 Hz, 1H), 3.12 (s, 6H), 2.44 (s, 3H).

[0060] Compounds 23, 24, 25, 28, 30, 33, 36, 39, 40, 42, 43, 44, 49, 50, 51, 54, 55, 56, 57, 58, 59, 60, 63, 65, 66, 87, 90, 91, 92, 96, and 101 were prepared in a manner similar to compound 27 with appropriate amines H and anilines D

##STR00026##

[0061] Among the listed reagents, solvents and catalysts in Scheme 3, SOCl.sub.2 is thionylchloride, RuPhos Pd G3 is (2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)[2-2′-amino-1,1′-biphenyl]] palladium(II) methanesulfonate, RuPhos is 2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl, and Cs.sub.2CO.sub.3 is cesium carbonate.

[0062] 4-Chloro-7-bromoquinazoline (M). To a suspension of 7-bromoquinazolin-4-ol L (1.17 g, 5.20 mmol) in dry PhMe (6 mL), SOCl.sub.2 (6 mL) and DMF (0.6 mL) were added. The resulting mixture was stirred at 95° C. for 9 h, cooled to room temperature, quenched with water (100 mL), and extracted with EtOAc (70 mL). The combined organic extracts were washed with water (200 mL) and brine (5 mL), dried over NA.sub.2SO.sub.4, and concentrated to afford the titled product M (1.26 g, 99%) as a yellow solid. LCMS (ESI) m/z calculated for C.sub.8H.sub.4BrClN.sub.2: 242, 244, 246; found: 243, 245, 247 [M+H].sup.+. .sup.1H-NMR (300 MHz, CDCl.sub.3): δ 9.05 (s, 1H), 8.28 (d, J=1.8 Hz, 1H), 8.15 (d, J=9.0 Hz, 1H), 7.84 (dd, J=9.0, 1.8 Hz, 1H).

[0063] 4-((7-Bromoquinazolin-4-yl)oxy)aniline (N). A mixture of 4-aminophenol (0.678 g, 6.21 mmol) and t-BuOK (0.668 g, 5.95 mmol) in dry THF (15 mL) at 0° C. was stirred for 20 min. Subsequently, 4-chloro-7-bromoquinazoline M (1.26 g, 5.17 mmol) was added slowly in small portions. The reaction mixture was then stirred for 3 h at 0° C., during which a suspension was formed. The suspension was filtered through a pad of celite. The pad was rinsed with THF (20 mL), and the resulting filtrate was evaporated to give a crude solid. The crude solid was suspended in MeOH (6 mL) and sonicated. Particles in the suspension were collected via filtration and dried to afford the titled product N (1.461 g, 89%) as an off-white solid. LCMS (ESI) m/z calculated for C.sub.14H.sub.10BrN.sub.3O: 315, 317; found: 316, 318 [M+H].sup.+.

[0064] 1-(4-((7-Bromoquinazolin-4-yl)oxy)phenyl)-3-((6-(trifluoromethyl)pyridin-3-yl)methyl)urea (O; compound 31). To a solution of 4-((7-Bromoquinazolin-4-yl)oxy)aniline N (305 mg, 0.964 mmol) and compound Q (461 mg, 1.35 mmol) in CH.sub.2Cl.sub.2 (6 mL), Et.sub.3N (0.27 mL) was added. The resulting mixture was refluxed for 1.5 d, during which a suspension was formed. Particles in the suspension was collected via filtration and rinsed with excess CH.sub.2Cl.sub.2 to afford the titled product O (462 mg, 93%) as a white solid. LCMS (ESI) m/z calculated for C.sub.22H.sub.15BrF.sub.3N.sub.5O.sub.2: 517, 519; found: 518, 520 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d6): δ 8.87 (s, 1H), 8.74 (s, 1H), 8.72 (br s, 1H), 8.30 (d, J=8.8 Hz, 1H), 8.24 (d, J=2.0 Hz, 1H), 8.00 (dd, J=8.0, 1.6 Hz, 1H), 7.93 (dd, J=8.8, 2.0 Hz, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.50 (AA′XX′, J.sub.AX=9.0 Hz, J.sub.AX′=0 Hz, 2H), 7.21 (AA′XX′, J.sub.AX=9.0 Hz, J.sub.AX′=0 Hz, 2H), 6.86 (t, J=6.0 Hz, 1H), 4.44 (d, J=6.0 Hz, 2H).

[0065] 1-(4-((7-(3,5-Dimethylpiperazin-1-yl)quinazolin-4-yl)oxy)phenyl)-3-((6-(trifluoromethyl)pyridin-3-yl)methyl)urea (P; compound 34). Freshly-distilled THF (4 mL) was purged with argon for 20 min, after which compound O (50 mg, 0.097 mmol), Cs.sub.2CO.sub.3 (47 mg, 0.15 mmol), cis-2,6-dimethylpiperazine (17 mg, 0.15 mmol), Ruphos (4.1 mg, 0.009 mmol), and Ruphos Pd G3 (4.1 mg, 0.005 mmol) were added to form a suspension. The resulting mixture was stirred under argon for 5 min. Subsequently, the reaction mixture was refluxed overnight. The resulting pale-yellow solution was filtered through a pad of celite and the filtrate was concentrated to give a residue. The residue was purified by column chromatography to afford the titled product P (49 mg, 92%) as a white solid. LCMS (ESI) m/z calculated for C.sub.28H.sub.28F.sub.3N.sub.7O.sub.2: 551; found: 552 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.85 (s, 1H), 8.72 (d, J=1.6 Hz, 1H), 8.46 (s, 1H), 8.09 (d, J=9.2 Hz, 1H), 8.00 (dd, J=8.0, 1.6 Hz, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.51 (dd, J=9.2, 2.4 Hz, 1H), 7.47 (AA′XX′, J.sub.AX=9.2 Hz, J.sub.AX′=0 Hz, 2H), 7.17-7.13 (m, 3H), 6.85 (t, J=5.6 Hz, 1H), 4.44 (d, J=5.6 Hz, 2H), 3.91-3.88 (m, 2H), 2.87-2.82 (m, 2H), 2.39-2.33 (m, 2H), 1.06 (d, J=6.4 Hz, 6H).

[0066] 1-(4-((7-(4-hydroxypiperidin-1-yl)quinazolin-4-yl)oxy)phenyl)-3-((6-(trifluoromethyl)pyridin-3-yl)methyl)urea (P; compound 30). Similar to compound 34, compound 30 was prepared from compound 31 and was obtained as a white solid. LCMS (ESI) m/z calculated for C.sub.28H.sub.28F.sub.3N.sub.7O.sub.2: 538; found: 539 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.82 (br s, 1H), 8.72 (d, J=1.2 Hz, 1H), 8.45 (s, 1H), 8.08 (d, J=9.2 Hz, 1H), 8.00 (dd, J=8.0, 1.2 Hz, 1H), 7.89 (d, J=8.0 Hz, 1H), 7.51-7.45 (m, 3H), 7.15-7.11 (m, 3H), 6.84 (t, J=6.0 Hz, 1H), 4.76 (d, J=4.0 Hz, 2H), 4.44 (d, J=6.0 Hz, 2H), 3.87-3.81 (m, 2H), 3.77-3.72 (m, 1H), 3.21-3.14 (m, 1H), 1.87-1.83 (m, 2H), 1.51-1.43 (m, 2H).

[0067] Compounds 24, 96, 97, 98, 99, 100 were prepared analogously to compound 34.

##STR00027##

[0068] Among the listed reagents, solvents, and catalysts listed in Scheme 3, POCl.sub.3 is phosphoryl chloride, CuI is copper iodide, 1,10-phen is 1,10-phenanthroline, K.sub.2CO.sub.3 is potassium carbonate, and tBuOH is tert-butanol.

[0069] 7-(Dimethylamino)quinazolin-4(3H)-one (R). A solution of 7-fluoroquinazolin-4(3H)-one A (1.65 g, 10.1 mmol) and dimethylamine (2M in THF, 20 mL, 40.2 mmol) in 2-methoxyethanol (60 mL) in a 250 mL sealed tube was stirred at 130° C. for 1 d, during which a solid was formed. Subsequently, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The resulting residue was suspended in MeOH (8 mL) and filtered to afford the titled product R (1.5 g, 79%) as a brown solid. LCMS (ESI) m/z calculated for C.sub.10H.sub.11N.sub.3O: 189; found: 190 [M+H].sup.+. .sup.1H NMR (300 MHz, DMSO-d.sub.6): δ 11.69 (br, 1H), 7.91 (s, 1H), 7.88 (d, J=9.0 Hz, 1H), 6.93 (dd, J=9.0, 2.4 Hz, 1H), 6.98 (d, J=2.4 Hz, 1H), 3.04 (s, 6H).

[0070] 4-Chloro-N,N-dimethylquinazolin-7-amine (S). To a suspension of 7-(dimethylamino)quinazolin-4(3H)-one R (910 mg, 4.81 mmol) in dry PhMe (10 mL), POCl.sub.3 (4 mL) was added, after which the resulting mixture was stirred at 95° C. for 12 h. The reaction mixture was the cooled to 0° C., quenched with ice water (100 mL), and neutralized with saturated sodium carbonate solution until the color of the solution turned from bright orange to pale yellow. Subsequently, the reaction mixture was extracted with EtOAc (170 mL), and the organic extract was washed with water (200 mL) and brine (20 mL), dried over NA.sub.2SO.sub.4, and concentrated to afford the titled product S (948 mg, 95%) as a yellow solid. LCMS (ESI) m/z calculated for C.sub.10H.sub.10ClN.sub.3: 207, 209; found: 208, 210 [M+H].sup.+. .sup.1H-NMR (300 MHz, CDCl.sub.3): δ 8.77 (s, 1H), 8.03 (d, J=9.3 Hz, 1H), 7.20 (dd, J=9.3, 2.7 Hz, 1H), 6.96 (d, J=2.7 Hz, 1H), 3.18 (s, 1H).

[0071] 4-((1H-indol-5-yl)oxy)-N,N-dimethylquinazolin-7-amine (T). DMF (4 mL) was purged with argon. Subsequently, 4-Chloro-N,N-dimethylquinazolin-7-amine S (322 mg, 1.55 mmol), 5-hydroxyindole (413 mg, 3.10 mmol), K.sub.2CO.sub.3 (429 mg, 3.10 mmol), CuI (29.5 mg, 0.155 mmol), and 1,10-phen (27.9 mg, 0.155 mmol) were added to form a reaction mixture, which was stirred at 90° C. for 3 h under argon. The reaction mixture was then diluted with EtOAc (15 mL) and water (15 mL), and filtered through a pad of celite. The filtrate was extracted with EtOAc (60 mL), and the organic extract was washed with water (120 mL) and brine (5 mL), dried over NA.sub.2SO.sub.4, and concentrated. The residue was suspended in MeOH and particles were collected by filtration to afford the titled product T (381 mg, 81%) as a white solid. LCMS (ESI) m/z calculated for C.sub.18H.sub.16N.sub.4O: 304; found: 305 [M+H].sup.+.

[0072] 5-((7-(dimethylamino)quinazolin-4-yl)oxy)-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)-1H-indole-1-carboxamide (U; compound 67). To a solution of 4-(4-amino-3-methoxyphenoxy)-N,N-dimethylquinazolin-7-amine T (170 mg, 0.95 mmol) in dry THF (5 mL), NaH (60% in oil, 112 mg, 2.79 mmol) was added. The resulting mixture was stirred at room temperature for 20 min. Subsequently, the mixture was cooled to −60° C., and 4-nitrophenyl ((6-(trifluoromethyl)pyridin-3-yl)methyl)carbamate V (574 mg, 1.68 mmol) was added in 3 portions. The mixture was then slowly warmed to 0° C., stirred for 1 h, and quenched with saturated ammonium chloride solution. The solvent was removed under reduced pressure and the resulting residue was diluted with EtOAc (35 mL), washed with water (80 mL) and brine (3 mL), dried over NA.sub.2SO.sub.4, and concentrated. The crude residue was purified by column chromatography to afford the titled product U (171 mg, 65%) as a white solid. LCMS (ESI) m/z calculated for C.sub.26H.sub.21F.sub.3N.sub.6O.sub.2: 506; found: 507 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.94 (t, J=5.6 Hz, 1H), 8.82 (d, J=1.6 Hz, 1H), 8.41 (s, 1H), 8.27 (d, J=8.8 Hz, 1H), 8.15 (d, J=9.2 Hz, 1H), 8.10 (dd, J=8.0, 1.6 Hz, 1H), 7.96 (d, J=3.6 Hz, 1H), 7.92 (d, J=8.0 Hz, 1H), 7.50 (d, J=2.4 Hz, 1H), 7.31 (dd, J=9.2, 2.4 Hz, 1H) 7.16 (dd, J=8.8, 2.4 Hz, 1H), 6.88 (d, J=2.4 Hz, 1H), 6.74 (d, J=3.6 Hz, 1H), 4.64 (d, J=5.6 Hz, 2H), 3.13 (s, 6H).

[0073] Compounds 29, 35, 38, 64, 68, 69, 83, and 89 were prepared in a manner similar to compound 67.

TABLE-US-00001 TABLE 1 Exemplary heterocyclic compounds Com- Calculated Mass pound Structure Mass [M + H].sup.+  1 [00028] 455 456  2 [00029] 469 470  3 [00030] 454 455  4 [00031] 399 400  5 [00032] 469 470  6 [00033] 469 470  7 [00034] 434 435  8 [00035] 374 375  9 [00036] 425 426  10 [00037] 469 470  11 [00038] 482 483  12 [00039] 443 444  13 [00040] 467 468  14 [00041] 417 418  15 [00042] 433 434  16 [00043] 433 434  17 [00044] 503 504  18 [00045] 417 418  19 [00046] 433 434  20 [00047] 503 504  21 [00048] 429 430  22 [00049] 415 416  23 [00050] 565 566  24 [00051] 552 553  25 [00052] 516 517  26 [00053] 498 499  27 [00054] 482 483  28 [00055] 537 538  29 [00056] 510 511  30 [00057] 538 539  31 [00058] 517 518  32 [00059] 457 458  33 [00060] 516 517  34 [00061] 551 552  35 [00062] 510 511  36 [00063] 500 501  37 [00064] 498 499  38 [00065] 496 497  39 [00066] 526 527  40 [00067] 512 513  41 [00068] 501 502  42 [00069] 512 513  43 [00070] 552 553  44 [00071] 539 540  45 [00072] 595 596  46 [00073] 551 552  47 [00074] 496 497  48 [00075] 495 496  49 [00076] 492 493  50 [00077] 419 420  51 [00078] 413 414  52 [00079] 449 450  53 [00080] 467 468  54 [00081] 414 415  55 [00082] 473 474  56 [00083] 481 482  57 [00084] 497 498  58 [00085] 427 428  59 [00086] 443 444  60 [00087] 438 439  61 [00088] 499 500  62 [00089] 503 504  63 [00090] 403 404  64 [00091] 457 458  65 [00092] 428 429  66 [00093] 444 445  67 [00094] 506 507  68 [00095] 507 508  69 [00096] 507 508  71 [00097] 398 399  72 [00098] 420 421  73 [00099] 481 482  74 [00100] 495 496  75 [00101] 506 507  76 [00102] 471 472  77 [00103] 558 559  78 [00104] 503 504  79 [00105] 481 482  80 [00106] 506 507  81 [00107] 464 465  82 [00108] 483 484  83 [00109] 507 508  84 [00110] 417 418  85 [00111] 519 520  86 [00112] 519 520  87 [00113] 409 410  88 [00114] 507 508  89 [00115] 507 508  90 [00116] 393 394  91 [00117] 434 435  92 [00118] 421 422  93 [00119] 428 429  94 [00120] 429 430  95 [00121] 443 444  96 [00122] 551 552  97 [00123] 565 566  98 [00124] 579 580  99 [00125] 524 525 100 [00126] 551 552 101 [00127] 420 421 102 [00128] 480 481 103 [00129] 496 497 104 [00130] 463 465 464 466 105 [00131] 463 465 464 466 106 [00132] 417 418 107 [00133] 385 386 108 [00134] 581 582

EXAMPLE 2: CSF1R KINASE ASSAY

[0074] A study was conducted to test certain compounds described in EXAMPLE 1 in inhibiting CSF1R kinase activity. Results from this study are shown in Table 2 below (see column 2).

[0075] Activity of CSF1R kinase was determined using a CSF1R Kinase-Glo assay. Recombinant N-terminal GST-CSF1R (CSF1R residues L534-C972) containing the CSFIR kinase domain was expressed in Sf9 insect cells and purified. The kinase assay was carried out in 96-well plates at 30° C. for 180 min with the tested compounds in a final volume of 50 μl including the following components: 25 mM Tris-HCl pH 7.4, 4 mM MnCl.sub.2, 10 mM MgCl.sub.2, 0.01% BSA, 0.5 mM NA.sub.3VO.sub.4, 0.02% Triton X-100, 40 μM ATP, 2 mM DTT and 20 μM poly(Glu,Tyr) 4:1 peptide, and 600 ng recombinant GST-CSF1R. Following incubation, 50 μl Kinase-Glo Plus Reagent (Promega, Madison, Wis., USA) was added and the mixture was incubated at 25° C. for 20 min. A 70-μL aliquot of each reaction mixture was transferred to a black microtiter plate and luminescence was measured on Wallac Vector 1420 multilabel counter (PerkinElmer, Shelton, Conn., USA). Percentage inhibition values were obtained by comparison of reaction rate with the rates in control wells (i.e., in the absence of test compound). IC.sub.50 values were calculated from a series of percentage inhibition values determined at a range of test compound concentrations using GraphPad Prism version 6 software (GraphPad, San Diego, Calif., USA).

EXAMPLE 3: CELLULAR PROLIFERATION ASSAYS

[0076] Studies were conducted to evaluate in vivo anticancer activity of certain compounds described in EXAMPLE 1 using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) cell viability assays. Results from these studies are shown in Table 2 below (see columns 3 and 4).

Cell line and culture:

[0077] The cell lines M-NFS-60 (ATCC®CRL-1838™) and BaF3-CSF1R-1600 were obtained from American Type Culture Collection (ATCC, Manassas, Va., USA). The stable BaF3-CSF1R-1600 cell line expresses ETV6-CSF1R fusion protein consisting of N-terminal ETS-variant gene 6 protein (ETV6 residues M1-G337) and CSF1R tyrosine kinase (CSF1R residues L533-C972). The M-NSF-60 and BaF3-CSF1R-1600 cells were cultured in RPMI1640 medium supplemented with 10% fetal bovine serum, 0.05 mM 2-ME, 10 U/ml penicillin, and 10 g/ml streptomycin at 37° C. and 5% CO.sub.2.

MTS cell viability assays:

[0078] M-NFS-60 and BaF3-CSF1R-1600 cells were seeded in 96-well plates at a density of 10000 cells/100 μl and 8000 cells/100 μl per well, respectively, for 16 h and treated with vehicle or various concentrations of test compounds in medium for 72 h. Viable cells were quantified using the MTS method (Promega, Madison, Wis., USA) according to manufacturer's recommended protocol. The results were determined by measuring absorbance at 490 nm using a plate reader (Victor 2). The GI50 value was defined as the amount of compound that caused 50% reduction in cell viability in comparison with DMSO-treated (vehicle) control and was calculated using Prism GraphPad Prism version 6 software (GraphPad).

TABLE-US-00002 TABLE 2 Inhibitory activity of heterocyclic compounds in a CSF1R kinase assay and in M-NFS-60 and BaF3-CSF1R cell assays Enzymatic Assay Cellular CSF1R Proliferation Assay enzyme M-NFS-60 (nM) BaF3-CSF1R Compound IC.sub.50, (nM) GI.sub.50 (nM) GI.sub.50 (nM)  1 +++ ++ ++  2 +++ +++ +++  3 +++ +++ +++  4 +++ +++ +++  5 +++ ++  6 +++  7 +  8 ++  9 +++ +++ +++  10 +++ ++ ++  11 +++ +++ +++  12 +++  13 ++  14 ++++ ++ ++++  15 ++++ ++ +++  16 ++++ +++ +++  17 ++++ +++ +++  18 ++++ ++ +++  19 ++++ +++ ++++  20 ++++ +++ +++  21 ++++ +++ +++  22 ++  23 +++ +++ +++  24 ++++ +++ +++  25 +++ + ++  26 +++ ++ +++  27 ++++ +++ +++  28 +++ +++ ++++  29 ++  30 ++++ +++ +++  31 ++ + ++  32 +++ + ++  33 +++ ++ ++  34 ++++ ++ ++++  35 +  36 +++ ++  37 ++  38 +  39 ++++ ++  40 +++ ++ +++  41 +++ ++  42 ++++ ++ +++  43 +++ ++ ++  44 ++++ ++ +++  45 ++++ ++ +++  47 +  49 ++++ + ++  50 +++  51 ++++ +++ +++  52 ++  53 ++  54 ++  55 +++  56 +++ ++ +++  57 ++++ ++ ++  58 ++++ ++ +++  59 ++++ +++ +++  60 +++  61 ++  62 ++++ ++  63 +++ ++ ++  64 +  65 ++++ +++ +++  66 ++++ +++ +++  67 +++ ++ +++  68 +++ ++ +++  69 +  71 ++  72 +  73 +++  75 +  77 ++  79 ++++  83 ++  84 ++  85 +++  87 +  89 ++  90 ++  93 +++  97 +++ +++ +++  98 ++++ +++ ++++  99 ++++ +++ +++ 100 ++++ +++ +++ 101 +++ ++ ++ 102 +++ + ++ 104 +++ ++ +++ 105 +++ +++ +++ 106 ++++ ++ + 107 +++ 108 +++ +++ +++ .sup.a ++++: IC.sub.50 < 20 nM; +++: 20 nM < IC.sub.50 < 100 nM; ++: 100 nM < IC.sub.50 < 1000 nM; +: .sup.b ++++: GI.sub.50 < 20 nM; +++: 20 nM < GI.sub.50 < 100 nM; ++: 100 nM < GI.sub.50 < 1000 nM; +:

EXAMPLE 4. KINASE SELECTIVITY PROFILE

[0079] A study was conducted to determine kinase selectivity of compounds 27 and 67. More specifically, each compound was tested for inhibitory activity of CSF1R kinase, as compared to that of seven other kinases, i.e., Aurora A, Aurora B, tyrosine-protein kinase Kit (c-Kit), fms-like tyrosine kinase 3 (FLT3), platelet-derived growth factor receptor (PDGFR) A, PDGFR B, and discoidin domain receptor tyrosine kinase 1 (DDR1). Results from this study are shown in Table 3 below.

[0080] In vitro profiling of the kinase panel was performed at Reaction Biology Corporation (www.reactionbiology.com, Malvern, Pa.) using the “HotSpot” assay platform. Briefly, specific kinase/substrate pairs along with required cofactors were prepared in a reaction buffer containing 20 mM Hepes pH 7.5, 10 mM MgCl.sub.2, 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM NA.sub.3VO.sub.4, 2 mM DTT, and 1% DMSO. Compound 27 or 67 was added. 20 minutes later, a mixture of ATP (Sigma, St. Louis Mo.) and .sup.33P ATP (Perkin Elmer, Waltham Mass.) was added to the resulting reaction solution to give a final concentration of 10 μM. The reaction was carried out at room temperature for 120 min., followed by spotting the reaction solution onto P81 ion exchange filter paper (Whatman Inc., Piscataway, N.J.). Unbound phosphate was removed by extensive washing filters in 0.75% phosphoric acid. After subtracting the background derived from a control reaction containing an inactive enzyme, kinase activity was determined as the percentage of the remaining kinase activity in a test sample compared to a vehicle (DMSO) reaction. An IC.sub.50 value and a dose-response curve for each compound against each kinase were obtained using Prism (Graph Pad Software). Selectivity, expressed as an IC.sub.50 ratio, was determined by dividing the IC.sub.50 value of a kinase, e.g., Aurora A, by that of CSF1R.

TABLE-US-00003 TABLE 3 Kinase selectivity profile Kinase IC.sub.50 (nM) Compound CSF1R Aurora A Aurora B c-Kit FLT3 PDGFR A PDGFR B DDR1 27 0.534 ++++ +++ ++ +++ ++ ++ +++ 67 5.69 +++ +++ +++ +++ ++ +++ + ++++: IC.sub.50 ratio > 1000; +++: 1000 > IC.sub.50 ratio > 100; ++: 100 > IC.sub.50 ratio > 10; +: 10 > IC.sub.50 ratio > 1

OTHER EMBODIMENTS

[0081] All of the features disclosed in this specification may be combined in any combination. Each feature disclosed in this specification may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is only an example of a generic series of equivalent or similar features.

[0082] Further, from the above description, one skilled in the art can easily ascertain the essential characteristics of the present invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions. Thus, other embodiments are also within the claims.