Compounds and methods for treating cancer

Abstract

Disclosed herein are compounds, compositions, and methods for treating cancer, inhibiting PARP1 and / or ATR, and / or the repair of DNA.

Description

[0001] COMPOUNDS AND METHODS FOR TREATING CANCER RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U. S. Provisional Application No.

[0003] 63 / 713,919, filed October 30, 2024, the contents of which are incorporated herein by reference in their entirety.

[0004] STATEMENT OF GOVERNMENT SUPPORT

[0005] This invention was made with government support under DE030445 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0006] BACKGROUND PARP1 and PARP2 are two key enzymes that mediate DNA damage responses (DDR) by serving as DNA damage sensors and signal transducers. To respond to DNA damage, such as nicks and double-strand breaks (DSB), PARP1 is rapidly recruited to the sites of the damaged DNA, and its catalytic activity increases 10- to 500-fold through an allosteric activation mechanism. This results in the synthesis of protein-conjugated poly(ADP-ribose) (PAR) chains using NAD+as a critical substrate. The negatively charged PARP functions as a high density protein binding scaffold and recruits components of the DNA damage repair machinery. Due to the rapid cellular division that is inherent to their nature, cancer cells are especially vulnerable to the inhibition of DDR mechanisms.

[0007] PARP inhibitors (PARPi) have been used to successfully treat breast and ovarian cancers with BRCA gene mutations, using synthetic lethal screening. Furthermore, PARPis have also been used to treat cancer patients with homologous recombination defects. However, acquired resistance to PARPi treatments impedes optimal clinical outcome. PARPi resistance may be obtained through increased expression or activity of replication fork stabilizers. For example, in PARPi-resistant cancer cells, the ataxia-telangiectasia-mutated-and-Rad3-related kinase (ATR) / checkpoint kinase 1 (CHK1) pathway is often upregulated, thereby inducing the phosphorylation of multiple proteins that stabilize the replication fork and HR repair. Moreover, common chemotherapeutic drugs, such as cisplatin, induce drug resistance through activating DNA damage response. To overcome such resistance, patients can be treated with a combination of PARPi and ATR inhibitors.

[0008] Therefore, dual inhibitors of PARP and ATR are thus an attractive target for cancer therapy. SUMMARY OF THE INVENTION

[0009] In certain aspects, the present disclosure provides compounds having a structure represented by Formula I, or a pharmaceutically acceptable salt thereof:

[0010]

[0011] I

[0012] wherein:

[0013] A is heterocyclyl or heteroaryl;

[0014] ml is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0015] nl is 1, 2, 3, 4, or 5;

[0016] X1is O, S, or NR10;

[0017] X2is O, S, or NR11;

[0018] X3is O, S, or NR12;

[0019] each of R1, R2, R3, R4, R5, R6, R7and R8is independently selected from H, alkoxy (e.g., methoxy or ethoxy), aralkyloxy (e.g., benzyloxy), fluoro, chloro, bromo, and iodo; each of R10, R11, and R12is independently selected from H, alkyl, and aralkyl; and

[0020] each R9is independently selected from H, halo, hydroxyl, amino, amido, alkyl, alkenyl, alkynyl, carboxyl, aryl, acetyl, ester, thioester, alkoxy, cyano, nitro, azido, alkylthio, cycloalkyl, heteroaryl, heterocyclylalkyl, and heterocyclyl.

[0021] In some aspects, the present disclosure provides pharmaceutical compositions comprising a compound of the present disclosure and a pharmaceutically acceptable excipient.

[0022] In certain aspects, the present disclosure provides methods of treating a cancer in a subject in need thereof, comprising administering to the subject an amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof. In some aspects, the present disclosure provides methods of inhibiting repair of DNA in a subject in need thereof, comprising administering to the subject an amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0023] In some aspects, the present disclosure provides methods of inhibiting PARP and ATR in a subject in need thereof, comprising administering to the subject an amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0024] BRIEF DESCRIPTION OF THE DRAWINGS FIGs. 1A-1J show the effect of JCS162-188 on cell viability. FIG. 1A shows cell viability after treatment with 0.25 / 0.5 / 1 / 2 pM JCS162 / 163 for 72 h in PC-3 and SUM149PT-BRCAlmut cells; FIG. IB shows cell viability after treatment with 0.5 pM or 1 pM JCS164 / 165 for 72 h in PC-3 and SUM149PT-BRCAlmut cells; FIG. 1C shows cell viability after treatment with 0.5 pM or 1 pM JCS166-169 for 72 h in PC-3 and SUM149PT-BRCAlmutcells; FIG. ID shows cell viability after treatment with 0.5 pM or 1 pM JCS170-173 for 72 h in PC-3 and SUM149PT-BRCAlmut cells; FIG. IE shows cell viability after treatment with 0.25 pM or 0.5 pM JCS174-177 for 72 h in PC-3 and SUM149PT-BRCAlmutcells; FIG. IF shows cell viability after treatment with 0.25 pMor0.5 pM JCS178-180 for 72 h inPC-3 and SUM149PT-BRCAlmut cells; FIG. 1G shows cell viability after treatment with 0.3 pM or 0.6 pM JCS181-184 for 72 h in PC-3 and SUM149PT-BRCAlmut cells; FIG. 1H shows cell viability after treatment with 0.3 pM or 0.6 pM JCS185-188 for 72 h in PC-3 and SUM149PT-BRCAlmut cells; FIG. II shows cell viability after treatment with 0.5 or 1 pM JCS 136 / 164 / 165 / 169 for 72 h in PC-3 cells; FIG. 1J shows cell viability after treatment with 0.4 or 0.8 pM JCS 189-192 for 72 h in PC-3 and SUM149PT-BRCAlmutcells.

[0025] FIGs.2A-2B show the effect of JCS 164 / 165 / 169 / 178 on cell viability in ovarian cancer cells. FIG.2A shows cell viability after treatment with 0.4 pM or 0.8 pM JCS 164 / 165 / 169 / 178 for 72 h in cisplatin-resistant OVCAR / CR cells; FIG. 2B shows cell viability after treatment with 0.5 / 1 / 2 pM JCS136 / 164 for 72 h in cisplatin-sensitive SKOV3 and cisplatin-resistant SKOV3 / CR cells.

[0026] FIG. 3 shows the effect of JCS 164 / 165 on cell viability in myeloma cells. In particular, FIG. 3 shows cell viability after treatment with 0.125 / 0.25 / 0.5 pM JCS164 / 165 for 72 h in RPMI 8226 and MM. IS cells, respectively.

[0027] FIGs. 4A-4I show JCS161-188 inhibits ATR activity in PC3 cells. FIG. 4A shows Olaparib induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 1 pM and 2 pM JCS 161-163; FIG. 4B shows Olaparib induced phosphorylation of Rad 17 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.5 pM and 1 pM JCS164 / 165; FIG. 4C shows Olaparib or UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.5 pM and 1 pM JCS166-169; FIG. 4D shows Olaparib or UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.5 pM and 1 pM JCS170-173; FIG. 4E shows Olaparib or UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.5 pM and 1 pM JCS174-177; FIG. 4F shows Olaparib or UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.25 ~ 1 pM JCS 178- 180; FIG. 4G shows Olaparib or UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.3 ~ 1 pM JCS181-184; FIG. 4H shows Olaparib or UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.3 ~ 1 pM JCS185-188; FIG. 41 shows Olaparib or UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.3 ~ 1 pM JCS189-192.

[0028] FIGs. 5A-5D show that the compounds of this disclosure inhibit ATR activity in ovarian cancer cells. FIG. 5A shows Olaparib induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.5 or 1 pM JCS 136 / 164 / 165 / 169 in OVCAR / CR cells; FIGs.5B, 5C, and 5D showUV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 1 pM JCS136 or JCS164 in SKOV3, A2780, and CP70 cells, respectively.

[0029] FIGs. 6A-6B show that the compounds of this disclosure inhibit ATR activity in myeloma cells. FIGs. 6A and 6B show UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.25 or 0.5 pM JCS164 / 165 in RPMI 8226 or MM.1S cells, respectively.

[0030] FIGs. 7A-7D show JCS141-188 trap PARP1 on DNA lesions in PC3 cells. FIG. 7A shows that PARP1 was trapped on DNA lesions by 0.5 pM JCS141-163; FIG. 7B shows that PARP1 was trapped on DNA lesions by 0.25 pM JCS 164- 173; FIG. 7C shows that PARP1 was trapped on DNA lesions by 0.5 pM JCS174-183; FIG. 7D shows that PARP1 was trapped on DNA lesions by 0.5 pM JCS 184-188.

[0031] FIGs. 8A-8C show that the compounds of this disclosure inhibit the growth of Olaparib-resistant breast cancer PDX. FIG. 8A shows tumor weight after the treatment with 60 or 100 mg / kg i.g. of JC099, or 60 mg / kg i.g. of JCS 144 / 152 / 164 / 165; FIG.8B shows tumor volume after the treatment with 60 or 100 mg / kg i.g. of JC099, or 60 mg / kg i.g. of JCS 144 / 152 / 164 / 165; FIG. 8C shows the body weight of mice after the treatment with 60 or 100 mg / kg i.g. of JC099, or 60 mg / kg i.g. of JCS 144 / 152 / 164 / 165. (One-way ANOVA, * P < 0.05; ** < 0.01).

[0032] FIGs. 9A-9C show that the compounds of this disclosure inhibit the growth of castration-resistant prostate cancer PDX. FIG.9A shows tumor weight after the treatment with 60 or 100 mg / kg i.g. of JC099, or 60 mg / kg i.g. of JCS164 / 165 / 169; FIG. 9B shows tumor volume after the treatment with 60 or 100 mg / kg i.g. of JC099, or 60 mg / kg i.g. of JCS 164 / 165 / 169; FIG. 9C shows the body weight of mice after the treatment with 60 or 100 mg / kg i.g. of JC099, or 60 mg / kg i.g. of JCS164 / 165 / 169. (One-way ANOVA, * P < 0.05; ** < 0.01; *** < 0.001).

[0033] FIGs. 10A-10B show the effect of JCS193-200 on cell viability. FIG. 10A shows cell viability after treatment with 0.4 or 0.8 pM JCS193-200 for 72 h in SUM149PT-BRCAlmut cells. FIG. 10B shows cell viability after treatment with 0.4 or 0.8 pM JCS 193-200 for 72 h in PC-3 cells.

[0034] FIGs. 11A-11E show that JCS193-200 inhibits ATR activity in PC3 cells. FIG. 11A shows Olaparib induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.3 or 0.6 pM JCS193-196; FIG. 11B shows UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.5 or 1 pM JCS193-196; FIG.

[0035] 11C shows Olaparib induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.3 or 0.6 pM JCS197-199; FIG. 11D shows UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.5 or 1 pM JCS197-199; FIG. 11E shows Olaparib induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites is inhibited by 0.4 pM JCS 136 / 164 / 169 / 178 / 193-200

[0036] FIG. 12 shows that JCS189-200 trap PARP1 on DNA lesions in PC3 cells. PARP1 was trapped on DNA lesions by 0.8 pM JCS 164 or JCS 189-200.

[0037] FIGs. 13A-13C show that JCS 164 / 169 / 178 / 194 inhibitthe growth of cisplatin-resistant ovarian cancer CDX. FIG. 13A shows tumor weight after the treatment with 60 mg / kg i.g. of Olaparib or JCS 164 / 169 / 178 / 194. FIG. 13B shows tumor volume after the treatment with 60 mg / kg i.g. of Olaparib or JCS 164 / 169 / 178 / 194. FIG. 13C shows the body weight of mice after the treatment with 60 mg / kg i.g. of Olaparib or JC S 164 / 169 / 178 / 194. (One-way ANOVA, * P < 0.05; ** < 0.01)

[0038] FIGs. 14A-14C show that JCS 169 / 178 / 193 / 199 inhibitthe growth of cisplatin-resistant ovarian cancer CDX. FIG. 14A shows tumor weight after the treatment with 60 mg / kg i.g. of Olaparib or JCS 169 / 178 / 193 / 199. FIG. 14B shows tumor volume after the treatment with 60 mg / kg i.g. of Olaparib or JCS JCS 169 / 178 / 193 / 199. FIG. 14C shows the body weight of mice after the treatment with 60 mg / kg i.g. of Olaparib or JCS JCS169 / 178 / 193 / 199. (One-way ANOVA, * P < 0.05; ** P < 0.01)

[0039] DETAILED DESCRIPTION OF THE INVENTION

[0040] In certain aspects, the present disclosure provides compounds having a structure represented by Formula I, or a pharmaceutically acceptable salt thereof:

[0041]

[0042] I

[0043] wherein:

[0044] A is heterocyclyl or heteroaryl;

[0045] ml is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0046] nl is 1, 2, 3, 4, or 5;

[0047] X1is O, S, or NR10;

[0048] X2is O, S, or NR11;

[0049] X3is O, S, or NR12;

[0050] each of R1, R2, R3, R4, R5, R6, R7and R8is independently selected from H, alkoxy (e.g., methoxy or ethoxy), aralkyloxy (e.g., benzyloxy), fluoro, chloro, bromo, and iodo; each of R10, R11, and R12is independently selected from H, alkyl, and aralkyl; and

[0051] each R9is independently selected from H, halo, hydroxyl, amino, amido, alkyl, alkenyl, alkynyl, carboxyl, aryl, acetyl, ester, thioester, alkoxy, cyano, nitro, azido, alkylthio, cycloalkyl, heteroaryl, heterocyclylalkyl, and heterocyclyl.

[0052] In certain embodiments, the compound is not:

[0053]

[0054] In some embodiments, the compound has a structure represented by Formula la, or a pharmaceutically acceptable salt thereof:

[0055]

[0056] la.

[0057] In certain such embodiments, R1is alkoxy (e.g., methoxy or ethoxy). In other embodiments, R1is aralkyloxy (e.g., benzyloxy). In some preferred embodiments, R1is fluoro. In some preferred embodiments, R2is fluoro. In certain preferred embodiments, R3is alkoxy e.g., methoxy or ethoxy). In certain preferred embodiments, R3is fluoro. In some embodiments, R3is chloro. In some embodiments, R4is H. In some preferred embodiments, R4is fluoro.

[0058] In some embodiments, R5is H. In certain preferred embodiments, R5is fluoro. In certain preferred embodiments, R6is fluoro. In other embodiments, R6is chloro. In some preferred embodiments, R7is fluoro. In some preferred embodiments, R8is fluoro. In other preferred embodiments, R8is alkoxy e.g., methoxy or ethoxy). In yet other embodiments, R8is aralkyloxy e.g., benzyloxy). In certain embodiments, nl is 1. In other embodiments, nl is 2.

[0059] In certain preferred embodiments, the compound has a structure represented by Formula lb, or a pharmaceutically acceptable salt thereof:

[0060]

[0061] In certain especially preferred embodiments, A is heterocyclyl (e.g., pyrrolidine or N-alkylpyrrolidine). In some embodiments, A is heteroaryl (e.g., imidazolyl or pyrazolyl). In some embodiments, ml is 1 or 2.

[0062] In certain preferred embodiments, A is selected from

[0063]

[0064]

[0065] wherein '* indicates the point of attachment.

[0066] In some embodiments, R12is H. In other embodiments, R12is alkyl e.g., methyl). In certain preferred embodiments, the compound has a structure represented by Formula Ic, or a pharmaceutically acceptable salt thereof:

[0067]

[0068] In some embodiments, the compound has a structure represented by Formula Id, or a pharmaceutically acceptable salt thereof:

[0069]

[0070] In some embodiments, the compound has a structure represented by Formula le, or a pharmaceutically acceptable salt thereof:

[0071]

[0072] In certain embodiments, the compound has a structure represented by Formula If, or a pharmaceutically acceptable salt thereof:

[0073]

[0074] In some embodiments, the compound has a structure represented by Formula Ig, or a pharmaceutically acceptable salt thereof:

[0075]

[0076] In certain embodiments, the compound has a structure represented by Formula Ih, or a pharmaceutically acceptable salt thereof:

[0077]

[0078] In certain embodiments, the compound has a structure represented by Formula li, or a pharmaceutically acceptable salt thereof:

[0079]

[0080] In some embodiments, the compound has a structure represented by Formula Ij, or a pharmaceutically acceptable salt thereof:

[0081]

[0082] In certain embodiments, the compound has a structure represented by Formula Ik, or a pharmaceutically acceptable salt thereof:

[0083]

[0084] In certain embodiments, the compound has a structure represented by Formula II, or a pharmaceutically acceptable salt thereof:

[0085]

[0086] II.

[0087] In certain embodiments, R9is alkyl (e.g., methyl or trifluorom ethyl). In other embodiments, R9is amino e.g., methylamino). In some embodiments, R9is H.

[0088] In some embodiments, the compound is selected from:

[0089]

[0090]

[0091]

[0092]

[0093] acceptable salt thereof.

[0094] In some aspects, the present disclosure provides pharmaceutical compositions comprising a compound of the present disclosure and a pharmaceutically acceptable excipient.

[0095] In certain aspects, the present disclosure provides methods of treating a cancer in a subject in need thereof, comprising administering to the subject an amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0096] In some embodiments, the cancer is breast cancer, head and neck cancer, lung cancer, prostate cancer, or ovarian cancer. In some embodiments, the cancer is testicular cancer, cervical cancer, bladder cancer, esophageal cancer, mesothelioma, or brain cancer (e.g., neuroblastoma).

[0097] In certain embodiments, the cancer is castration-resistant. In some such embodiments, the cancer is castration-resistant prostate cancer (CRPC). In certain embodiments, the cancer is relapsed. In some embodiments, the cancer is refractory. In some embodiments, the cancer is resistant to treatment with olaparib. In some embodiments, the cancer is resistant to treatment with cisplatin. In certain embodiments, the cancer is resistant to treatment with enzalutamide.

[0098] In some aspects, the present disclosure provides methods of inhibiting repair of DNA in a subject in need thereof, comprising administering to the subject an amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0099] In some aspects, the present disclosure provides methods of inhibiting PARP and ATR in a subject in need thereof, comprising administering to the subject an amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0100] Pharmaceutical Compositions

[0101] The compositions and methods of the present invention may be utilized to treat an individual in need thereof. In certain embodiments, the individual is a mammal such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or the compound is preferably administered as a pharmaceutical composition comprising, for example, a compound of the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil, or injectable organic esters. In preferred embodiments, when such pharmaceutical compositions are for human administration, particularly for invasive routes of administration (i.e., routes, such as injection or implantation, that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free, or substantially pyrogen-free. The excipients can be chosen, for example, to effect delayed release of an agent or to selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form such as tablet, capsule (including sprinkle capsule and gelatin capsule), granule, lyophile for reconstitution, powder, solution, syrup, suppository, injection or the like. The composition can also be present in a transdermal delivery system, e.g., a skin patch. The composition can also be present in a solution suitable for topical administration, such as a lotion, cream, or ointment. A pharmaceutically acceptable carrier can contain physiologically acceptable agents that act, for example, to stabilize, increase solubility or to increase the absorption of a compound such as a compound of the invention. Such physiologically acceptable agents include, for example, carbohydrates, such as glucose, sucrose or dextrans, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier, including a physiologically acceptable agent, depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition can be a selfemulsifying drug delivery system or a selfmicroemulsifying drug delivery system. The pharmaceutical composition (preparation) also can be a liposome or other polymer matrix, which can have incorporated therein, for example, a compound of the invention. Liposomes, for example, which comprise phospholipids or other lipids, are nontoxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer.

[0102] The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0103] The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as com starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations. A pharmaceutical composition (preparation) can be administered to a subject by any of a number of routes of administration including, for example, orally (for example, drenches as in aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); absorption through the oral mucosa (e.g., sublingually); subcutaneously; transdermally (for example as a patch applied to the skin); and topically (for example, as a cream, ointment or spray applied to the skin). The compound may also be formulated for inhalation. In certain embodiments, a compound may be simply dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions suitable for same can be found in, for example, U. S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, as well as in patents cited therein.

[0104] The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 1 percent to about ninety-nine percent of active ingredient, preferably from about 5 percent to about 70 percent, most preferably from about 10 percent to about 30 percent.

[0105] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound of the invention, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0106] Formulations of the invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), lyophile, powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and / or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient. Compositions or compounds may also be administered as a bolus, electuary or paste.

[0107] To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents, such as, modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.

[0108] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surfaceactive or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0109] The tablets, and other solid dosage forms of the pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. They may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

[0110] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophiles for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, cyclodextrins and derivatives thereof, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, com, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

[0111] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0112] Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0113] Dosage forms for the topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that may be required.

[0114] The ointments, pastes, creams and gels may contain, in addition to an active compound, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof. Powders and sprays can contain, in addition to an active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0115] Transdermal patches have the added advantage of providing controlled delivery of a compound of the present invention to the body. Such dosage forms can be made by dissolving or dispersing the active compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

[0116] The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

[0117] Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0118] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

[0119] In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

[0120] Injectable depot forms are made by forming microencapsulated matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissue.

[0121] For use in the methods of this invention, active compounds can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.

[0122] Methods of introduction may also be provided by rechargeable or biodegradable devices. Various slow release polymeric devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including proteinaceous biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form an implant for the sustained release of a compound at a particular target site.

[0123] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0124] The selected dosage level will depend upon a variety of factors including the activity of the particular compound or combination of compounds employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound(s) being employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound(s) employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0125] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the pharmaceutical composition or compound at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. By “therapeutically effective amount” is meant the concentration of a compound that is sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the weight, sex, age, and medical history of the subject. Other factors which influence the effective amount may include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent being administered with the compound of the invention. A larger total dose can be delivered by multiple administrations of the agent. Methods to determine efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison’s Principles of Internal Medicine 13 ed., 1814-1882, herein incorporated by reference).

[0126] In general, a suitable daily dose of an active compound used in the compositions and methods of the invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

[0127] If desired, the effective daily dose of the active compound may be administered as one, two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. In certain embodiments of the present invention, the active compound may be administered two or three times daily. In preferred embodiments, the active compound will be administered once daily.

[0128] The patient receiving this treatment is any animal in need, including primates, in particular humans; and other mammals such as equines, cattle, swine, sheep, cats, and dogs; poultry; and pets in general.

[0129] In certain embodiments, compounds of the invention may be used alone or conjointly administered with another type of therapeutic agent. The present disclosure includes the use of pharmaceutically acceptable salts of compounds of the invention in the compositions and methods of the present invention. In certain embodiments, contemplated salts of the invention include, but are not limited to, alkyl, dialkyl, trialkyl or tetra-alkyl ammonium salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, L-arginine, benethamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, IH-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, l-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, l-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, 1-ascorbic acid, 1-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfuric acid, ethane-l,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptonic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, 1-malic acid, malonic acid, mandelic acid, methanesulfonic acid, naphthal ene-l,5-disulfonic acid, naphthal ene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, proprionic acid, 1 -pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, 1-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid acid salts.

[0130] The pharmaceutically acceptable acid addition salts can also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such solvate can be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or adventitious to such solvent.

[0131] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0132] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal-chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0133] Definitions

[0134] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings that are commonly understood by those of ordinary skill in the art. Generally, nomenclature used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics and protein and nucleic acid chemistry, described herein, are those well known and commonly used in the art.

[0135] The methods and techniques of the present disclosure are generally performed, unless otherwise indicated, according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout this specification. See, e.g. “Principles of Neural Science”, McGraw-Hill Medical, New York, N. Y. (2000); Motulsky, “Intuitive Biostatistics”, Oxford University Press, Inc. (1995); Lodish et al., “Molecular Cell Biology, 4th ed.”, W. H. Freeman & Co., New York (2000); Griffiths et al., “Introduction to Genetic Analysis, 7th ed ”, W. H. Freeman & Co., N. Y. (1999); and Gilbert et al., “Developmental Biology, 6th ed ”, Sinauer Associates, Inc., Sunderland, MA (2000).

[0136] Chemistry terms used herein, unless otherwise defined herein, are used according to conventional usage in the art, as exemplified by “The McGraw-Hill Dictionary of Chemical Terms”, Parker S., Ed., McGraw-Hill, San Francisco, C. A. (1985).

[0137] All of the above, and any other publications, patents and published patent applications referred to in this application are specifically incorporated by reference herein. In case of conflict, the present specification, including its specific definitions, will control.

[0138] The term “agent” is used herein to denote a chemical compound (such as an organic or inorganic compound, a mixture of chemical compounds), a biological macromolecule (such as a nucleic acid, an antibody, including parts thereof as well as humanized, chimeric and human antibodies and monoclonal antibodies, a protein or portion thereof, e.g., a peptide, a lipid, a carbohydrate), or an extract made from biological materials such as bacteria, plants, fungi, or animal (particularly mammalian) cells or tissues. Agents include, for example, agents whose structure is known, and those whose structure is not known.

[0139] A “patient,” “subject,” or “individual” are used interchangeably and refer to either a human or a non-human animal. These terms include mammals, such as humans, primates, livestock animals (including bovines, porcines, etc.), companion animals (e.g., canines, felines, etc.) and rodents (e.g., mice and rats).

[0140] “Treating” a condition or patient refers to taking steps to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment.

[0141] The term “preventing” is art-recognized, and when used in relation to a condition, such as a local recurrence (e.g., pain), a disease such as cancer, a syndrome complex such as heart failure or any other medical condition, is well understood in the art, and includes administration of a composition which reduces the frequency of, or delays the onset of, symptoms of a medical condition in a subject relative to a subject which does not receive the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a population of patients receiving a prophylactic treatment relative to an untreated control population, and / or delaying the appearance of detectable cancerous growths in a treated population versus an untreated control population, e.g., by a statistically and / or clinically significant amount.

[0142] “Administering” or “administration of’ a substance, a compound or an agent to a subject can be carried out using one of a variety of methods known to those skilled in the art. For example, a compound or an agent can be administered, intravenously, arterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and transdermally (by absorption, e.g., through a skin duct). A compound or agent can also appropriately be introduced by rechargeable or biodegradable polymeric devices or other devices, e.g., patches and pumps, or formulations, which provide for the extended, slow or controlled release of the compound or agent. Administering can also be performed, for example, once, a plurality of times, and / or over one or more extended periods.

[0143] Appropriate methods of administering a substance, a compound or an agent to a subject will also depend, for example, on the age and / or the physical condition of the subject and the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability and toxicity). In some embodiments, a compound or an agent is administered orally, e.g., to a subject by ingestion. In some embodiments, the orally administered compound or agent is in an extended release or slow release formulation, or administered using a device for such slow or extended release.

[0144] As used herein, the phrase “conjoint administration” refers to any form of administration of two or more different therapeutic agents such that the second agent is administered while the previously administered therapeutic agent is still effective in the body (e.g., the two agents are simultaneously effective in the patient, which may include synergistic effects of the two agents). For example, the different therapeutic compounds can be administered either in the same formulation or in separate formulations, either concomitantly or sequentially. Thus, an individual who receives such treatment can benefit from a combined effect of different therapeutic agents.

[0145] A “therapeutically effective amount” or a “therapeutically effective dose” of a drug or agent is an amount of a drug or an agent that, when administered to a subject will have the intended therapeutic effect. The full therapeutic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses. Thus, a therapeutically effective amount may be administered in one or more administrations. The precise effective amount needed for a subject will depend upon, for example, the subject’s size, health and age, and the nature and extent of the condition being treated, such as cancer or MDS. The skilled worker can readily determine the effective amount for a given situation by routine experimentation.

[0146] As used herein, the terms “optional” or “optionally” mean that the subsequently described event or circumstance may occur or may not occur, and that the description includes instances where the event or circumstance occurs as well as instances in which it does not. For example, “optionally substituted alkyl” refers to the alkyl may be substituted as well as where the alkyl is not substituted.

[0147] It is understood that substituents and substitution patterns on the compounds of the present invention can be selected by one of ordinary skilled person in the art to result chemically stable compounds which can be readily synthesized by techniques known in the art, as well as those methods set forth below, from readily available starting materials. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon or on different carbons, so long as a stable structure results.

[0148] As used herein, the term “optionally substituted” refers to the replacement of one to six hydrogen radicals in a given structure with the radical of a specified substituent including, but not limited to: hydroxyl, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH2-O-alkyl, -OP(O)(O-alkyl)2 or -CH2-OP(O)(O-alkyl)2. Preferably, “optionally substituted” refers to the replacement of one to four hydrogen radicals in a given structure with the substituents mentioned above. More preferably, one to three hydrogen radicals are replaced by the substituents as mentioned above. It is understood that the substituent can be further substituted.

[0149] As used herein, the term “alkyl” refers to saturated aliphatic groups, including but not limited to C1-C10 straight-chain alkyl groups or C1-C10 branched-chain alkyl groups. Preferably, the “alkyl” group refers to Ci-Ce straight-chain alkyl groups or Ci-Ce branched-chain alkyl groups. Most preferably, the “alkyl” group refers to C1-C4 straight-chain alkyl groups or C1-C4 branched-chain alkyl groups. Examples of “alkyl” include, but are not limited to, methyl, ethyl, 1 -propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, 1 -pentyl, 2-pentyl, 3 -pentyl, neo-pentyl, 1 -hexyl, 2-hexyl, 3 -hexyl, 1 -heptyl, 2-heptyl, 3 -heptyl, 4-heptyl, 1 -octyl, 2-octyl, 3-octyl or 4-octyl and the like. The “alkyl” group may be optionally substituted.

[0150] The term “acyl” is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0151] The term “acylamino” is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula hydrocarbylC(O)NH-.

[0152] The term “acyloxy” is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0153] The term “alkoxy” refers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy and the like.

[0154] The term “alkoxyalkyl” refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl.

[0155] The term “alkyl” refers to saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In preferred embodiments, a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., Ci-30 for straight chains, C3-30 for branched chains), and more preferably 20 or fewer.

[0156] Moreover, the term “alkyl” as used throughout the specification, examples, and claims is intended to include both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, etc.

[0157] The term “Cx-y” or “Cx-Cy”, when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups that contain from x to y carbons in the chain. Coalkyl indicates a hydrogen where the group is in a terminal position, a bond if internal. A Ci-ealkyl group, for example, contains from one to six carbon atoms in the chain.

[0158] The term “alkylamino”, as used herein, refers to an amino group substituted with at least one alkyl group.

[0159] The term “alkylthio”, as used herein, refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

[0160] The term “amido”, as used herein, refers to a group

[0161]

[0162] wherein R9and R10each independently represent a hydrogen or hydrocarbyl group, or R9and R10taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

[0163] The terms “amine” and “amino” are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, e.g., a moiety that can be represented by

[0164]

[0165] wherein R9, R10, and R10’ each independently represent a hydrogen or a hydrocarbyl group, or R9and R10taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

[0166] The term “aminoalkyl”, as used herein, refers to an alkyl group substituted with an ammo group. The term “aralkyl”, as used herein, refers to an alkyl group substituted with an aryl group.

[0167] The term “aralkyloxy”, as used herein, refers to an alkyloxy group substituted on the alkyl group with an aryl group. A representative aralkyloxy group is benzyloxy, -O-CH2-C6H5.

[0168] The term “aryl” as used herein include substituted or unsubstituted single-ring aromatic groups in which each atom of the ring is carbon. Preferably the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term “aryl” also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0169] The term “carbamate” is art-recognized and refers to a group

[0170]

[0171] wherein R9and R10independently represent hydrogen or a hydrocarbyl group.

[0172] The term “carbocyclylalkyl”, as used herein, refers to an alkyl group substituted with a carbocycle group.

[0173] The term “carbocycle” includes 5-7 membered monocyclic and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated and aromatic rings. Carbocycle includes bicyclic molecules in which one, two or three or more atoms are shared between the two rings. The term “fused carbocycle” refers to a bicyclic carbocycle in which each of the rings shares two adjacent atoms with the other ring. Each ring of a fused carbocycle may be selected from saturated, unsaturated and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits, is included in the definition of carbocyclic. Exemplary “carbocycles” include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-lH-indene and bicyclo[4.1.0]hept-3-ene. “Carbocycles” may be substituted at any one or more positions capable of bearing a hydrogen atom. The term “carbocyclylalkyl”, as used herein, refers to an alkyl group substituted with a carbocycle group.

[0174] The term “carbonate” is art-recognized and refers to a group -OCO2-.

[0175] The term “carboxy”, as used herein, refers to a group represented by the formula -CO2H.

[0176] The term “cycloalkyl” includes substituted or unsubstituted non-aromatic single ring structures, preferably 4- to 8-membered rings, more preferably 4- to 6-membered rings. The term “cycloalkyl” also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is cycloalkyl and the substituent (e.g., R100) is attached to the cycloalkyl ring, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, denzodioxane, tetrahydroquinoline, and the like.

[0177] The term “ester”, as used herein, refers to a group -C(O)OR9wherein R9represents a hydrocarbyl group.

[0178] The term “ether”, as used herein, refers to a hydrocarbyl group linked through an oxygen to another hydrocarbyl group. Accordingly, an ether substituent of a hydrocarbyl group may be hydrocarbyl-O-. Ethers may be either symmetrical or unsymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include “alkoxyalkyl” groups, which may be represented by the general formula alkyl-O-alkyl.

[0179] The terms “halo” and “halogen” as used herein means halogen and includes chloro, fluoro, bromo, and iodo.

[0180] The terms “hetaralkyl” and “heteroaralkyl”, as used herein, refers to an alkyl group substituted with a hetaryl group.

[0181] The terms “heteroaryl” and “hetaryl” include substituted or unsubstituted aromatic single ring structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms “heteroaryl” and “hetaryl” also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like. The term “heteroatom” as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0182] The term “heterocyclylalkyl”, as used herein, refers to an alkyl group substituted with a heterocycle group.

[0183] The terms “heterocyclyl”, “heterocycle”, and “heterocyclic” refer to substituted or unsubstituted non-aromatic ring structures, preferably 3- to 10-membered rings, more preferably 3- to 7-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms “heterocyclyl” and “heterocyclic” also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heterocyclic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyl s. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

[0184] The term “hydrocarbyl”, as used herein, refers to a group that is bonded through a carbon atom that does not have a =0 or =S substituent, and typically has at least one carbonhydrogen bond and a primarily carbon backbone, but may optionally include heteroatoms. Thus, groups like methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered to be hydrocarbyl for the purposes of this application, but substituents such as acetyl (which has a =0 substituent on the linking carbon) and ethoxy (which is linked through oxygen, not carbon) are not. Hydrocarbyl groups include, but are not limited to aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

[0185] The term “hydroxyalkyl”, as used herein, refers to an alkyl group substituted with a hydroxy group.

[0186] The term “lower” when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups where there are ten or fewer atoms in the substituent, preferably six or fewer. A “lower alkyl”, for example, refers to an alkyl group that contains ten or fewer carbon atoms, preferably six or fewer. In certain embodiments, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein are respectively lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, whether they appear alone or in combination with other substituents, such as in the recitations hydroxyalkyl and aralkyl (in which case, for example, the atoms within the aryl group are not counted when counting the carbon atoms in the alkyl substituent).

[0187] The terms “polycyclyl”, “poly cycle”, and “polycyclic” refer to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls) in which two or more atoms are common to two adjoining rings, e.g., the rings are “fused rings”. Each of the rings of the polycycle can be substituted or unsubstituted. In certain embodiments, each ring of the poly cycle contains from 3 to 10 atoms in the ring, preferably from 5 to 7.

[0188] The term “sulfate” is art-recognized and refers to the group -OSO3H, or a pharmaceutically acceptable salt thereof.

[0189] The term “sulfonamido” is art-recognized and refers to the group represented by the general formulae

[0190]

[0191] wherein R9and R10independently represents hydrogen or hydrocarbyl.

[0192] The term “sulfoxide” is art-recognized and refers to the group-S(O)-.

[0193] The term “sulfonate” is art-recognized and refers to the group SO3H, or a pharmaceutically acceptable salt thereof.

[0194] The term “sulfone” is art-recognized and refers to the group -S(O)2-.

[0195] The term “substituted” refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include any substituents described herein, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.

[0196] The term “thioalkyl”, as used herein, refers to an alkyl group substituted with a thiol group.

[0197] The term “thioester”, as used herein, refers to a group -C(O)SR9or -SC(O)R9wherein R9represents a hydrocarbyl.

[0198] The term “thioether”, as used herein, is equivalent to an ether, wherein the oxygen is replaced with a sulfur.

[0199] The term “urea” is art-recognized and may be represented by the general formula

[0200]

[0201] wherein R9and R10independently represent hydrogen or a hydrocarbyl.

[0202] The term “modulate” as used herein includes the inhibition or suppression of a function or activity (such as cell proliferation) as well as the enhancement of a function or activity.

[0203] The phrase “pharmaceutically acceptable” is art-recognized. In certain embodiments, the term includes compositions, excipients, adjuvants, polymers and other materials and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0204] “Pharmaceutically acceptable salt” or “salt” is used herein to refer to an acid addition salt or a basic addition salt which is suitable for or compatible with the treatment of patients.

[0205] The term “pharmaceutically acceptable acid addition salt” as used herein means any non-toxic organic or inorganic salt of any base compounds represented by Formula I. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric and phosphoric acids, as well as metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids that form suitable salts include mono-, di-, and tricarboxylic acids such as glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, benzoic, phenylacetic, cinnamic and salicylic acids, as well as sulfonic acids such as p-toluene sulfonic and methanesulfonic acids. Either the mono or di-acid salts can be formed, and such salts may exist in either a hydrated, solvated or substantially anhydrous form. In general, the acid addition salts of compounds of Formula I are more soluble in water and various hydrophilic organic solvents, and generally demonstrate higher melting points in comparison to their free base forms. The selection of the appropriate salt will be known to one skilled in the art. Other non-pharmaceutically acceptable salts, e.g., oxalates, may be used, for example, in the isolation of compounds of Formula I for laboratory use, or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

[0206] The term “pharmaceutically acceptable basic addition salt” as used herein means any non-toxic organic or inorganic base addition salt of any acid compounds represented by Formula I or any of their intermediates. Illustrative inorganic bases which form suitable salts include lithium, sodium, potassium, calcium, magnesium, or barium hydroxide. Illustrative organic bases which form suitable salts include aliphatic, alicyclic, or aromatic organic amines such as methylamine, trimethylamine and picoline or ammonia. The selection of the appropriate salt will be known to a person skilled in the art.

[0207] Many of the compounds useful in the methods and compositions of this disclosure have at least one stereogenic center in their structure. This stereogenic center may be present in a R or a S configuration, said R and S notation is used in correspondence with the rules described in Pure Appl. Chem. (1976), 45, 11-30. The disclosure contemplates all stereoisomeric forms such as enantiomeric and diastereoisomeric forms of the compounds, salts, prodrugs or mixtures thereof (including all possible mixtures of stereoisomers). See, e.g., WO 01 / 062726.

[0208] Furthermore, certain compounds which contain alkenyl groups may exist as Z (zusammen) or E (entgegen) isomers. In each instance, the disclosure includes both mixture and separate individual isomers.

[0209] “Prodrug” or “pharmaceutically acceptable prodrug” refers to a compound that is metabolized, for example hydrolyzed or oxidized, in the host after administration to form the compound of the present disclosure (e.g., compounds of formula I). Typical examples of prodrugs include compounds that have biologically labile or cleavable (protecting) groups on a functional moiety of the active compound. Prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, or dephosphorylated to produce the active compound. Examples of prodrugs using ester or phosphorami date as biologically labile or cleavable (protecting) groups are disclosed in U. S. Patents 6,875,751, 7,585,851, and 7,964,580, the disclosures of which are incorporated herein by reference. The prodrugs of this disclosure are metabolized to produce a compound of Formula I. The present disclosure includes within its scope, prodrugs of the compounds described herein. Conventional procedures for the selection and preparation of suitable prodrugs are described, for example, in “Design of Prodrugs” Ed. H. Bundgaard, Elsevier, 1985.

[0210] The phrase “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material useful for formulating a drug for medicinal or therapeutic use.

[0211] The term “Log of solubility”, “LogS” or “logS” as used herein is used in the art to quantify the aqueous solubility of a compound. The aqueous solubility of a compound significantly affects its absorption and distribution characteristics. A low solubility often goes along with a poor absorption. LogS value is a unit stripped logarithm (base 10) of the solubility measured in mol / liter.

[0212] EXAMPLES

[0213] The invention now being generally described, it will be more readily understood by reference to the following examples which are included merely for purposes of illustration of certain aspects and embodiments of the present invention and are not intended to limit the invention.

[0214] Example 1: Synthesis of Exemplary Compounds of the Disclosure

[0215] General Procedure A:

[0216] The compounds were generally prepared by reaction of the corresponding aldehydes, e.g., 3,4,5-trifluorobenzaldehyde, with tert-butyl (4-oxocyclohexyl)carbamate in the presence of 0.4 equiv. cetyltrimethylammonium bromide and 2.5 equiv. sodium hydroxide to give tertbutyl (4-oxo-3,5-bis((£)-3,4,5-trifluorobenzylidene)cyclohexyl)carbamate. The Boc group was then deprotected by treatment of the carbamate with TFA. Benzoylation of the resulting 4-amino-2,6-bis((E)-3,4,5-trifluorobenzylidene)cyclohexan-l-one with 2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxylic acid using the standard peptide coupling reagents, TBTU or EDC and HOAt, afforded the A-(4-oxo-3,5-bis((E)-3,4,5- trifluorobenzylidene)cyclohexyl)-2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxamide, e.g., JCS164.

[0217] Synthesis of JCS164

[0218]

[0219] N-(4-Oxo-3,5-bis((E)-3,4,5-trifluorobenzylidene)cyclohexyl)-2-((2-(pyrrolidin-l-yl)ethyl)amino)pyri-midine-5-carboxamide (JCS164)

[0220] To a mixture of the A-tert-butyl (4-oxocyclohexyl)carbamate (213.28 mg, 1 mmol, 1.0 equiv.), cetyltrimethylammonium bromide (145 mg, 0.4 mmol, 0.4 equiv.), and water (5.0 mL) in a round bottom flask was added dropwise sodium hydroxide (100 mg, 2.5 mmol, 2.5 equiv.) in water (2 mL), and the reaction was stirred for 10 min at 60 °C. Then to this mixture was added 3,4,5-difluorobenzaldehyde (400.2 mg, 2.5 mmol, 2.5 equiv.). The reaction mixture was then allowed to stir at 60 °C for 12 h. The yellow precipitate thus obtained was filtered, washed with water and dried to get the pure product tert-butyl (4-oxo-3,5-bis((E)-3,4,5-trifluorobenzylidene)cyclohexyl)carbamate (447 mg, 90% yield).

[0221] Trifluoroacetic acid (0.5 ml) was added to a solution of tert-butyl (4-oxo-3,5-bis((E)-3,4,5-trifluorobenzylidene)cyclohexyl)carbamate (248.6 mg, 0.5 mmol) in dichloromethane (5.0 ml) at 21 °C and stirred overnight at 21 °C. The solvent of the reaction solution was distilled off under reduced pressure and the resulting residue was poured into a IN aqueous sodium hydroxide solution and extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain the crude product which was purified with silica gel chromatography with a solvent system of 10% MeOH / ethyl acetate to 100% MeOH to obtain 4-amino-2,6-bis((E)-3,4,5-trifluorobenzylidene)cyclohexan-l-one (317 mg, 80% yield) as a yellow solid.

[0222] To a stirred solution of 4-amino-2,6-bis((£)-3,4,5-trifluorobenzylidene)cyclohexan-l-one (397.3 mg, 1 mmol, 1.0 equiv.) in THF (2 mL) was added diisopropylethylamine (521 uL, 3 mmol, 3 equiv.) at 0 °C. Then, 2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxylic acid (236.2 mg, 1 mmol, 1.0 equiv.) in THF (3 mL) and 2-(177-benzotriazole-l-yl)-l, 1,3,3- tetramethylaminium tetrafluorob orate (TBTU) (481 mg, 1.5 mmol, 1.5 equiv.) was added and stirred at 21 °C for 12 h. The reaction solvent was evaporated under reduced pressure and washed with brine and extracted with ethyl acetate. Then the solvent was evaporated and the crude material was purified with silica gel chromatography with solvent system of 50% MeOH / ethyl acetate to 100% MeOH to obtain N-(4-oxo-3,5-bis((E)-3,4,5-trifluorobenzylidene)cyclo-hexyl)-2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxamide JCS164 (430 mg, 70% yield) as a yellow solid. The product was further crystallized from MeOH to obtain the pure compound.

[0223] Synthesis of 2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxylic acid

[0224]

[0225] To a microwave vial equipped with a stir bar was added methyl 2-chloropyrimidine-5-carboxylate (0.517 g, 3.0 mmol, 1.0 equiv.), 2-(pyrrolidin-l-yl)ethan-l -amine (0.85 g, 7.5 mmol, 2.5 equiv.), diisopropylethylamine (1.56 mL, 3 mmol, 3 equiv.) and acetonitrile (5 mL). The mixture was heated in a microwave at 80 °C for 24 h, after which time it was diluted with ethyl acetate (15 mL), washed with saturated aqueous sodium bicarbonate (10 mL) arid brine (10 mL), dried over sodium sulfate and evaporated to give a crude residue. The pure product was obtained by flash chromatography on silica gel (gradient elution 10% methanol / ethyl acetate- 100% methanol) to give the desired product, methyl 2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxylate (0.637 g, 85% yield), as a brown oil.

[0226] Methyl 2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxylate (0.75 g, 3.0 mmol, 1.0 equiv.) was dissolved in 15 mL ethanol and sodium hydroxide (0.6 g, 15.0 mmol, 5.0 equiv.) in water (5 mL) was added dropwise. The mixture was stirred at 60 °C for 5 h. The ethanol was removed in vacuo and the aqueous solution was acidified with cone. HC1 at 5 °C. The solid was collected, treated with methanol, filtered and dried at 55-60 °C in vacuo to provide 2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxylic acid (0.66 g, 92% yield) as a brown solid which was pure enough to be used for next step. The following compounds were synthesized by General Method A: JCS165, JCS166, JCS167, JCS168, JCS169, JCS170, JCS171, JCS172, JCS173, JCS174, JCS175, JCS176, JCS177, JCS178, JCS179, JCS180, JCS181, JCS182, JCS183, JCS184, JCS185, JCS186, JCS187, JCS188, JCS189, JCS190, JCS191, JCS192, JCS193, JCS194, JCS195, JCS196, JCS197, JCS198, JCS199, and JCS 200.

[0227]

[0228] JV-(4-Oxo-3,5-bis((E)-3,4,5-trifluorobenzylidene)cyclohexyl)-2-((2-(pyrrolidin-l-yl)ethyl)amino)pyri-midine-5-carboxamide (JCS164)

[0229] 'H NMR (500 MHz, CDC13) 58.61 (s, 2H), 7.70 (s, 2H), 7.07 - 7.03 (m, 4H), 6.36 (m, 1H), 6.15 (m, 1H), 4.51 -4.44 (m, 1H), 3.55 - 3.51 (m, 2H), 3.24 (br d, J= 14.8 Hz, 2H), 3.05 (dd, J= 15.6, 8.4 Hz, 2H), 2.71 (t, J= 6.1 Hz, 2H), 2.56 (t, J= 6.4 Hz, 4H), 1.79 - 1.77 (m, 4H).

[0230] 13C NMR (126 MHz, CDCI3) 6 187.3, 164.4, 163.1, 157.7 (4C), 151.3 (dd.., = 250.3 Hz, Jc-CF = 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 137.2 (2C), 133.6 (2C), 131.0 (dd, JC-C-CF = 6.3 Hz, Jc-C-CF = 3.7 Hz, 2C), 116.2, 114.5 (d, JC-CF = 20.1 Hz, 4C), 54.3, 53.9, 44.5, 40.2, 33.7, 23.6.

[0231] 19F NMR (376 MHz, CDCI3) 5 -133.0 (d, JFF = 20.2 Hz, 2F), -157.4 (t, JFF = 18.8 Hz, IF HR-APCI m / z calcd for C31H28F6N5O2 [M+H] = 616.2141, found 616.2119.

[0232]

[0233] 2-((((5)-l-Ethylpyrrolidin-2-yl)methyl)amino)-7V-(4-oxo-3,5-bis((E')-3,4,5-trifluorobenzylidene)cyclo-hexyl)pyrimidine-5-carboxamide (JCS165)

[0234] 'H NMR (500 MHz, CDC13) 6 8.60 (s, 2H), 7.71 (s, 2H), 7.07 - 7.04 (m, 4H), 6.20 (s, 1H), 6.07 (s, 1H), 4.50 - 4.46 (m, 1H), 3.68 - 3.62 (m, 1H), 3.34 - 3.16 (m, 4H), 3.05 (dd, J= 15.9, 8.3 Hz, 2H), 2.87 - 2.79 (m, 1H), 2.69 - 2.61 (m, 1H), 2.26 - 2.15 (m, 2H), 1.93 - 1.54 (m, 4H), 1.09 (t, J = 7.2 Hz, 3H).

[0235] 13C NMR (126 MHz, CDCE) 6 187.3, 164.4, 163.6, 157.7 (4C), 151.3 (dd. Ja = 250.3 Hz, Jc-CF = 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 137.2 (2C), 133.6 (2C), 131.0 (dd, Jc-C-CF = 6.3 Hz, Jc-C-CF = 3.7 Hz, 2C), 116.2, 114.5 (dd, JC-CF = 13.8 Hz, JC-CF = 3.7 Hz, 4C), 62.4, 53.6, 48.1, 44.5, 42.8, 33.7, 28.5, 22.8, 14.0.

[0236] 19F NMR (376 MHz, CDCb) 5 -133.0 (d, JFF = 22.5 Hz, 2F), -157.4 (t, JFF = 18.8 Hz, IF). HR-APCI m / z calcd for C32H30F6N5O2 [M+H] = 630.2298, found 630.2119.

[0237]

[0238] 2-((((5)-l-Methylpyrrolidin-2-yl)methyl)amino)-A-(4-oxo-3,5-bis((£')-3,4,5-trifluorobenzylidene)cy-clohexyl)pyrimidine-5-carboxamide (JCS166)

[0239] 'H NMR (500 MHz, CDCI3) 6 8.69 (s, 2H), 7.68 (s, 2H), 7.07 - 7.03 (m, 4H), 6.88 (s, 1H), 6.72 (s, 1H), 4.50 -4.36 (m, 1H), 3.76 - 3.68 (m, 1H), 3.50 - 3.43 (m, 1H), 3.25 (br d, J= 15.9 Hz, 2H), 3.03 (m, 2H), 2.87 - 2.79 (m, 1H), 2.47 (s, 3H), 2.45 - 2.37 (m, 1H), 2.04 - 1.65 (m, 4H).

[0240] 13C NMR (126 MHz, CDCI3) 6 187.4, 164.3, 163.4, 158.3 (2C), 157.7 (2C), 151.3 (dd, JCF = 250.3 Hz, JC-CF = H.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 136.8 (2C), 133.9 (2C), 131.0 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7Hz, 2C), 116.2, 114.5 (dd, JC-CF = 13.8 Hz, Jc. CF = 3.7 Hz, 4C), 64.9, 57.4, 44.7, 42.4, 40.7, 33.7, 28.4, 22.5.

[0241] 19F NMR (376 MHz, CDCb) 5 -133.2 (d, JFF = 22.3 Hz, 2F), -157.6 (t, JFF = 18.8 Hz, IF). HR-APCI m / z calcd for C31H28F6N5O2 [M+H] = 616.2141, found 616.2138.

[0242]

[0243] N-(3,5-bis((Zi)-3,4-Difluoro-5-methoxybenzylidene)-4-oxocyclohexyl)-2-((2-(pyrrolidin-l-yl)ethyl)-amino)pyrimidine-5-carboxamide (JCS167)

[0244] 'H NMR (500 MHz, CDC13) 58.62 (s, 2H), 7.67 (s, 2H), 6.84 - 6.73 (m, 4H), 6.69 (m, 1H), 6.55 (m, 1H), 4.50 - 4.42 (m, 1H), 3.88 (s, 6H), 3.53 - 3.48 (m, 2H), 3.23 (br d, J= 14.8 Hz, 2H), 3.05 (dd, J= 15.6, 8.4 Hz, 2H), 2.69 (t, J= 6.1 Hz, 2H), 2.55 (t, J= 6.4 Hz, 4H), 1.78 - 1.74 (m, 4H).

[0245] 13C NMR (126 MHz, CDCI3) 5 187.8, 164.4, 163.0, 157.8 (d, JC-F = 31.5 Hz, 2C), 150.9 (dd, JC-F = 248.2, 11.3 Hz, 2C), 149.2 (dd, JC-F = 8.8, 5.0 Hz, 2C), 141.4 (dd, JC-F = 246.9, 13.8 Hz, 2C), 138.4 (2C), 132.2 (2C), 130.6 (dd, JC-F = 8.8, 5.0 Hz, 2C), 116.3, 111.1 (2C), 110.5 (d, JC-F= 18.9 Hz, 2C), 56.8, 54.4, 53.9, 44.6, 40.1, 33.6, 23.5.

[0246] 19F NMR (376 MHz, CDCb) 5 -136.2 (d, JFF= 18.8 Hz, IF), -156.7 (d, JFF= 18.8 Hz, IF).

[0247] HR-APCI m / z calcd for C33H34F4N5O4 [M+H] = 640.2541, found 640.2530.

[0248]

[0249] 7V-(3,5-bis((Zi)-3,4-Difluoro-5-methoxybenzylidene)-4-oxocyclohexyl)-2-((((»y)-l-ethylpyrrolidin-2-yl)methyl)amino)pyrimidine-5-carboxamide (JCS168)

[0250] 'H NMR (500 MHz, CDCI3) 58.61 (s, 2H), 7.65 (s, 2H), 6.83 - 6.72 (m, 4H), 6.60 (m, 1H), 6.26 (m, 1H), 4.50 - 4.45 (m, 1H), 3.87 (s, 6H), 3.62 - 3.56 (m, 1H), 3.29 - 3.05 (m, 6H), 2.83 - 2.74 (m, 1H), 2.64- 2.58 (m, 1H), 2.24 -2.11 (m, 2H), 1.88 - 1.49 (m, 4H), 1.05 (t, J= 7.2 Hz, 3H).13C NMR (126 MHz, CDC13) 5 187.8, 164.4, 163.4, 157.8 (d, JC. F= 31.5 Hz, 2C), 150.9 (dd, JC-F = 248.2, 11.3 Hz, 2C), 149.2 (dd, JC-F = 8.8, 5.0 Hz, 2C), 141.4 (dd, JC-F = 246.9, 13.8 Hz, 2C), 138.4 (2C), 132.8 (2C), 130.6 (dd, JC-F = 8.8, 5.0 Hz, 2C), 116.2, 111.2 (2C), 110.5 (d, JC-F = 18.9 Hz, 2C), 62.3, 56.8, 53.5, 48.1, 44.5, 42.8, 33.5, 28.4, 22.7, 13.9.

[0251] 19F NMR (376 MHz, CDCb) 5 -136.2 (d, JFF = 18.8 Hz, IF), -156.5 (d, JFF = 18.8 Hz, IF).

[0252] HR-APCI m / z calcd for C34H36F4N5O4 [M+H] = 654.2697, found 654.2682.

[0253]

[0254] 7V-(3,5-bis((Zi)-3,4-Difluoro-5-methoxybenzylidene)-4-oxocyclohexyl)-2-((((»y)-l-methylpyrrolidin-2-yl)methyl)amino)pyrimidine-5-carboxamide (JCS169)

[0255] 'H NMR (500 MHz, CDCI3) 58.61 (s, 2H), 7.67 (s, 2H), 6.83 - 6.53 (m, 4H), 6.54 (m, 1H), 6.27 (m, 1H), 4.50 - 4.45 (m, 1H), 3.87 (s, 6H), 3.62 - 3.56 (m, 1H), 3.32 - 3.05 (m, 6H), 2.48 - 2.40 (m, 1H), 2.30 (s, 3H), 2.25 - 2.18 (m, 1H), 1.90- 1.51 (m, 4H).

[0256] 13C NMR (126 MHz, CDCI3) 5 187.8, 164.4, 163.4, 157.8 (d, JC-F = 31.5 Hz, 2C), 150.9 (dd, JC-F = 248.2, 11.3 Hz, 2C), 149.2 (dd, JC-F = 8.8, 5.0 Hz, 2C), 141.4 (dd, JC-F = 246.9, 13.8 Hz, 2C), 138.4 (2C), 132.7 (2C), 130.6 (dd, JC-F = 8.8, 5.0 Hz, 2C), 116.2, 111.2 (2C), 110.5 (d, JC-F = 18.9 Hz, 2C), 63.9, 57.3, 56.8, 44.5, 42.4, 40.4, 33.6, 28.4, 22.6.

[0257] 19F NMR (376 MHz, CDCb) 5 -136.2 (d, JFF = 18.8 Hz, IF), -156.5 (d, JFF = 18.8 Hz, IF).

[0258] HR-APCI m / z calcd for C34H36F4N5O4 [M+H] = 640.2697, found 640.2682.

[0259]

[0260] 7V-(3,5-bis((Zi)-3-Ethoxy-4,5-difluorobenzylidene)-4-oxocyclohexyl)-2-((((»y)-l-ethylpyrrolidin-2-yl)methyl)amino)pyrimidine-5-carboxamide (JCS170)

[0261] 'H NMR (500 MHz, CDCb) 58.61 (s, 2H), 7.65 (s, 2H), 6.83 - 6.72 (m, 4H), 6.57 (m, 1H), 6.30 (m, 1H), 4.54 - 4.47 (m, 1H), 4.08 (q, J= 7.0 Hz, 4H), 3.64 - 3.59 (m, 1H), 3.31 - 3.05 (m, 6H), 2.83 - 2.74 (m, 1H), 2.67 - 2.62 (m, 1H), 2.24 - 2.11 (m, 2H), 1.88 - 1.49 (m, 4H), 1.43 (t, J = 7.0 Hz, 6H), 1.05 (t, J= 7.2 Hz, 3H).

[0262] 13C NMR (126 MHz, CDCb) 5 187.8, 164.3, 163.4, 157.8 (d, JC-F = 31.5 Hz, 2C), 150.9 (dd, JC-F = 248.2, 11.3 Hz, 2C), 149.2 (dd, JC. F= 8.8, 5.0 Hz, 2C), 141.4 (dd, JC. F= 246.9, 13.8 Hz, 2C), 138.4 (2C), 132.8 (2C), 130.6 (dd, JC-F = 8.8, 5.0 Hz, 2C), 116.2, 112.3 (2C), 110.3 (d, JC. F= 18.9 Hz, 2C), 65.5, 62.3, 53.4, 48.0, 44.3, 42.7, 33.5, 28.3, 22.6, 14.6, 13.7

[0263] 19F NMR (376 MHz, CDCb) 5 -136.1 (d, JFF= 18.8 Hz, IF), -155.9 (d, JFF= 18.8 Hz, IF).

[0264] HR-APCI m / z calcd for C36H40F4N5O4 [M+H] = 682.3010, found 682.3014.

[0265]

[0266] 7V-(3,5-bis((Zi)-3-Ethoxy-4,5-difluorobenzylidene)-4-oxocyclohexyl)-2-((2-(pyrrolidin-l-yl)ethyl)ami-no)pyrimidine-5-carboxamide (JCS171)

[0267] 'H NMR (500 MHz, CDCb) 58.70 (s, 2H), 7.70 (s, 2H), 6.89 - 6.76 (m, 6H), 4.50 - 4.42 (m, 1H), 4.08 (q, J= 7.0 Hz, 4H), 3.68 - 3.63 (m, 2H), 3.23 (brd, J= 14.8 Hz, 2H), 3.10 (dd, J= 15.6, 8.4 Hz, 2H), 2.69 (t, J= 6.1 Hz, 2H), 2.55 (t, J = 6.4Hz, 4H), 1.78 - 1.74 (m, 4H), 1.45 (t, = 7.0 Hz, 6H).

[0268] 13C NMR (126 MHz, CDCb) 5 187.8, 164.1, 162.9, 157.8 (d, JC-F = 31.5 Hz, 2C), 150.9 (dd, JC-F = 248.2, 11.3 Hz, 2C), 149.2 (dd, JC-F = 8.8, 5.0 Hz, 2C), 141.4 (dd, JC-F = 246.9, 13.8 Hz, 2C), 138.1 (2C), 132.9 (2C), 130.6 (dd, JC-F = 8.8, 5.0 Hz, 2C), 116.4, 112.3 (2C), 110.5 (d, JC-F= 18.9 Hz, 2C), 65.5, 54.3, 54.0, 44.6, 39.7, 33.6, 23.4, 14.6.

[0269] 19F NMR (376 MHz, CDCb) 5 -136.3 (d, JFF= 26.3 Hz, IF), -156.1 (d, JFF= 26.3 Hz, IF).

[0270] HR-APCI m / z calcd for C35H38F4N5O4 [M+H] = 668.2854, found 668.2854.

[0271]

[0272] 7V-(3,5-bis((Zi)-3-(Benzyloxy)-4,5-difluorobenzylidene)-4-oxocyclohexyl)-2-((2-(pyrrolidin-l-yl)eth-yl)amino)pyrimidine-5-carboxamide (JCS172)

[0273] 'H NMR (500 MHz, DMSO-d6) 58.60 (s, 2H), 8.30 (s, 1H), 7.58 (s, 1H), 7.49 (s, 2H), 7.36 -7.08 (m, 14H), 5.16 (s, 4H), 4.01 - 3.93 (m, 1H), 3.35 - 3.19 (m, 4H), 3.02 (br d, J= 14.8 Hz, 2H), 2.80 (dd, J= 15.6, 8.4 Hz, 2H), 2.38 (t, J= 6.4 Hz, 4H), 1.57 - 1.52 (m, 4H).

[0274] 13C NMR (126 MHz, DMSO-d6) 5 187.5, 163.7, 162.7, 157.9 (d, JC-F = 31.5 Hz, 2C), 150.9 (dd, JC-F = 248.2, 11.3 Hz, 2C), 147.4 (dd, JC-F = 8.8, 5.0 Hz, 2C), 140.3 (dd, JC-F = 246.9, 13.8 Hz, 2C), 135.9 (2C), 135.7 (2C), 134.7 (2C), 131.2 (dd, JC-F = 8.8, 5.0 Hz, 2C), 128.5 (4C), 128.2 (2C), 127.9 (4C), 116.4, 113.2 (2C), 110.6 (d, JC-F = 18.9 Hz, 2C), 70.8, 54.4, 53.5 (2C), 44.7, 32.9, 23.0. One high-field carbon not observed.

[0275] 19F NMR (376 MHz, DMSO-d6) 5 -137.3 (d. JH= 18.8 Hz, IF), -157.7 (d. JH= 18.8 Hz, IF).

[0276] HR-APCI m / z calcd for C45H42F4N5O4 [M+H] = 792.3167, found 792.3176.

[0277]

[0278] 7V-(3,5-bis((E')-3-(Benzyloxy)-4,5-difluorobenzylidene)-4-oxocyclohexyl)-2-((((»y)-l-ethylpyrrolidin-2-yl)methyl)amino)pyrimidine-5-carboxamide (JCS173)

[0279] 'H NMR (500 MHz, DMSO-d6) 58.70 (br s, 2H), 8.46 (s, 1H), 7.79 (s, 1H), 7.61 (s, 2H), 7.49 - 7.19 (m, 14H), 5.28 (s, 4H), 4.12 - 4.06 (m, 1H), 3.60 - 3.56 (m, 1H), 3.46 - 3.22 (m, 6H), 3.14 (br d, J= 14.8 Hz, 2H), 2.98 (dd, J= 15.6, 8.4 Hz, 2H), 1.91 - 1.66 (m, 4H), 1.09 (t, J= 7.2 Hz, 3H).13C NMR (126 MHz, DMSO-d6) 5 188.0, 164.0, 163.3, 158.3 (d, JC-F = 31.5 Hz, 2C), 150.9 (dd, JC-F = 248.2, 11.3 Hz, 2C), 147.4 (dd, JC-F = 8.8, 5.0 Hz, 2C), 140.3 (dd, JC-F = 246.9, 13.8 Hz, 2C), 136.4 (2C), 131.1 (2C), 135.1 (2C), 131.2 (dd, JC-F = 8.8, 5.0 Hz, 2C), 129.0 (4C), 128.6 (2C), 128.4 (4C), 116.7, 113.7 (2C), 110.5 (d, JC-F = 18.9 Hz, 2C), 71.3, 53.6 (2C overlapped), 48.8, 45.2, 45.0, 33.4 (2C), 28.5, 22.6. One high-field carbon not observed.

[0280] 19F NMR (376 MHz, DMSO-d6) 5 -137.3 (d, JFF = 18.8 Hz, 2F), -157.7 (d, JFF = 18.8 Hz, 2F).

[0281] HR-APCI m / z calcd for C46H44F4N5O4 [M+H] = 806.3323, found 806.3331.

[0282]

[0283] 4-Methyl-V-(4-oxo-3,5-bis( E,)-3,4,5-trifluorobenzylidene)cyclohexyl)-2-((2-(pyrrolidin- l-yl)ethyl)-amino)pyrimidine-5-carboxamide (JCS174)

[0284] 'H NMR (500 MHz, CDC13) 58.21 (s, 1H), 7.70 (s, 2H), 7.07 - 7.03 (m, 4H), 6.07 (m, 1H), 5.93 (m, 1H), 4.49 - 4.41 (m, 1H), 3.55 - 3.46 (m, 2H), 3.22 - 3.08 (m, 4H), 2.68 (t, J= 6.1 Hz, 2H), 2.56 (t, J= 6.4 Hz, 4H), 2.38 (s, 3H), 1.79- 1.77 (m, 4H).

[0285] 13C NMR (126 MHz, CDCI3) 6 187.3, 166.8, 162.0, 156.8 (4C), 151.3 (dd. Ja = 250.3 Hz, Jc-CF= 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 137.3 (2C), 133.2 (2C), 130.9 (dd, Jc-C-CF = 6.3 Hz, Jc-C-CF = 3.7 Hz, 2C), 117.5, 114.4 (dd, JC-CF = 20.1 Hz, JC-CF = 7.5 Hz, 4C), 54.6, 53.9, 44.5, 40.1, 33.4, 23.6 (2C’s overlapped).

[0286] 19F NMR (376 MHz, CDCb) 5 -133.0 (d, JFF = 20.2 Hz, 2F), -157.4 (t, JFF = 18.8 Hz, IF). HR-APCI m / z calcd for C32H30F6N5O2 [M+H] = 630.2298, found 630.2302.

[0287]

[0288] 2-((((5)-l-Ethylpyrrolidin-2-yl)methyl)amino)-4-methyl-7V-(4-oxo-3,5-bis((E)-3,4,5-trifluorobenzyli-dene)cyclohexyl)pyrimidine-5-carboxamide (JCS175)

[0289] 'H NMR (500 MHz, CDC13) 58.18 (s, 1H), 7.66 (s, 2H), 7.05 - 7.02 (m, 4H), 6.16 (m, 1H), 5.91 (m, 1H), 4.49 - 4.43 (m, 1H), 3.63 - 3.56 (m, 1H), 3.26 - 3.09 (m, 6H), 2.84 - 2.74 (m, 1H), 2.62 -2.56 (m, 1H), 2.37 (s, 3H), 2.24 - 2.10 (m, 2H), 1.88 - 1.51 (m, 4H), 1.06 (t, J = 7.2 Hz, 3H).

[0290] 13C NMR (126 MHz, CDCI3) 6 187.4, 166.8, 162.4, 156.8 (4C), 151.3 (dd. Ja = 250.3 Hz, Jc-CF= 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 137.3 (2C), 133.5 (2C), 130.9 (dd, Jc-C-CF = 6.3 Hz, Jc-C-CF = 3.7 Hz, 2C), 117.4, 114.4 (dd, JC-CF = 20.1 Hz, JC-CF = 7.5 Hz, 4C), 62.5, 53.6, 48.1, 44.4, 42.7, 33.2, 28.4, 23.3, 22.8, 13.9.

[0291] 19F NMR (376 MHz, CDCb) 5 -133.0 (d, JFF = 20.2 Hz, 2 F), -157.4 (t, JFF = 18.8 Hz, IF).

[0292] HR-APCI m / z calcd for C33H32F6N5O2 [M+H] = 644.2454, found 644.2458.

[0293]

[0294] JV-(4-Oxo-3,5-bis((E)-3,4,5-trifluorobenzylidene)cyclohexyl)-2-((2-(pyrrolidin-l-yl)ethyl)amino)-4-(trifluoromethyl)pyrimidine-5-carboxamide (JCS176)

[0295] 'H NMR (500 MHz, CDCI3) 58.32 (s, 1H), 7.63 (s, 2H), 7.04 - 7.00 (m, 4H), 6.58 (s, 1H), 6.42 (s, 1H), 4.35 - 4.23 (m, 1H), 3.52 (t, J= 6.1 Hz, 2H), 3.18 (br d, J= 14.8 Hz, 2H), 2.95 (dd, J = 15.6, 8.4 Hz, 2H), 2.76 (t, J= 6.1 Hz, 2H), 2.60 (t, J= 6.4 Hz, 4H), 1.83 - 1.72 (m, 4H).

[0296] 13C NMR (126 MHz, CDCI3) 6 186.6, 164.8, 161.7, 159.8 (4C), 151.3 (dd. Ja = 250.3 Hz, Jc-CF= 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 137.0 (2C), 133.2 (2C), 130.8 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 115.9, 115.6, 114.4 (dd, JC-CF = 20.1 Hz, JC-CF = 7.5 Hz, 4C), 54.3, 53.6, 44.9, 39.8, 32.9, 23.3.

[0297] 19F NMR (376 MHz, CDCI3) 5 -66.1 (s, 3F), -133.0 (d, JFF = 20.2 Hz, 2F), -157.4 (t, JFF = 18.8 Hz, IF).

[0298] HR-APCI m / z calcd for C32H27F9N5O2 [M+H] = 684.2015, found 684.2016.

[0299]

[0300] 2-((((5)-l-Ethylpyrrolidin-2-yl)methyl)amino)-7V-(4-oxo-3,5-bis((E)-3,4,5-trifluorobenzylidene)-cyclohexyl)-4-(trifluoromethyl)pyrimidine-5-carboxamide (JCS177)

[0301] 'H NMR (500 MHz, CDCI3) 58.47 (s, 1H), 7.71 (s, 2H), 7.05 - 7.02 (m, 4H), 6.39 (m, 1H), 6.09 (m, 1H), 4.49 - 4.43 (m, 1H), 3.69 - 3.64 (m, 1H), 3.38 - 3.06 (m, 6H), 2.87 - 2.67 (m, 2H), 2.28 -2.19 (m, 2H), 1.94- 1.57 (m, 4H), 1.11 (t, J= 7.2Hz, 3H).

[0302] 13C NMR (126 MHz, CDCI3) 6 186.6, 164.3, 162.1, 160.4 (4C), 151.3 (dd.., = 250.3 Hz, Jc-CF = 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 137.5 (2C), 132.9 (2C), 130.8 (dd, JC-C-CF = 6.3 Hz, Jc-C-CF = 3.7 Hz, 2C), 116.3, 116.1, 114.3 (dd, JC-CF = 20.1 Hz, JC-CF = 7.5 Hz, 4C), 62.7, 53.4, 48.4, 44.8, 42.6, 32.9, 28.3, 22.7, 13.6.

[0303] 19F NMR (376 MHz, CDCI3) 5 -66.0 (s, 3F), -133.1 (d, JFF = 20.2 Hz, 2F), -157.4 (t, JFF = 18.8 Hz, IF).

[0304] HR-APCI m / z calcd for C33H29F9N5O2 [M+H] = 698.2172, found 698.2174.

[0305]

[0306] A-Methyl-A-(4-oxo-3,5-bis((£')-3,4,5-trifluorobenzylidene)cyclohexyl)-2-((2-(pyrrolidin- l-yl)ethyl)-amino)pyrimidine-5-carboxamide (JCS178)

[0307] 'H NMR (500 MHz, CDC13) 58.39 (s, 2H), 7.67 (s, 2H), 7.07 - 6.99 (m, 4H), 6.14 (ms, 1H), 4.58 - 4.44 (m, 1H), 3.57 - 3.48 (m, 2H), 3.17 - 3.02 (m, 7H), 2.76 - 2.68 (m, 2H), 2.56 (t, J= 6.4 Hz, 4H), 1.79 - 1.77 (m, 4H).

[0308] 13C NMR (126 MHz, CDCI3) 6 186.9, 168.3, 162.2, 158.1 (4C), 151.3 (dd.., = 250.3 Hz, Jc-CF= 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 136.3 (2C), 134.3 (2C), 130.9 (dd, JC-C-CF = 6.3 Hz, Jc-C-CF = 3.7 Hz, 2C), 118.0, 114.2 (d, JC-CF = 20.1 Hz, 4C), 54.3, 53.8 (2C), 39.9, 31.0, 23.5 (2C). Two high-field carbons not observed.

[0309] 19F NMR (376 MHz, CDCb) 5 -133.0 (d, JFF = 20.2 Hz, 2F), -157.6 (t, JFF = 18.8 Hz, IF). HR-APCI m / z calcd for C32H30F6N5O2 [M+H] = 630.2298, found 630.2298.

[0310]

[0311] 4-(Methylamino)-7V-(4-oxo-3,5-bis((E)-3,4,5-trifluorobenzylidene)cyclohexyl)-2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxamide (JCS179)

[0312] 'H NMR (500 MHz, CDCI3) 58.40 (br s, 1H), 8.07 (br s, 1H), 7.61 (s, 2H), 7.03 - 7.00 (m, 4H), 6.60 (br s, 1H), 5.90 (br s, 1H), 4.49 - 4.41 (m, 1H), 3.48 - 3.44 (m, 2H), 3.20 (br d, J= 14.8 Hz, 2H), 3.06 (dd, J= 15.6, 8.4 Hz, 2H), 2.90 (s, 3H), 2.63 (t, J= 6.1 Hz, 2H), 2.51 (t, J=6.4Hz, 4H), 1.79 - 1.77 (m, 4H).13C NMR (126 MHz, CDCb) 6 187.7, 167.6, 162.2, 155.7 (4C), 151.3 (dd. Ja = 250.3 Hz, Jc-CF = 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 136.7 (2C), 133.9 (2C), 130.9 (dd, JC-C-CF = 6.3 Hz, Jc-C-CF = 3.7 Hz, 2C), 114.4 (dd, JC-CF = 20.1 Hz, JC-CF = 7.5 Hz, 4C), 98.4, 54.7, 53.9, 44.0, 40.0, 33.4, 26.8, 23.4.

[0313] 19F NMR (376 MHz, CDCb) 5 -133.2 (d, JFF= 20.2 Hz, 3F), -157.7 (t, JFF= 18.8 Hz, IF). HR-APCI m / z calcd for C32H31F6N6O2 [M+H] = 645.2407, found 645.2412.

[0314]

[0315] 2-((((5)-l-Ethylpyrrolidin-2-yl)methyl)amino)-4-(methylamino)-A-(4-oxo-3,5-bis((£')- 3,4,5-trifluoro-benzylidene)cyclohexyl)pyrimidine-5-carboxamide (JCS180)

[0316] 'H NMR (500 MHz, CDCb) 58.43 (m, 1H), 8.06 (br s, 1H), 7.64 (s, 2H), 7.05 - 7.02 (m, 4H), 6.39 (m, 1H), 5.70 (m, 1H), 4.49 - 4.43 (m, 1H), 3.69 - 3.64 (m, 1H), 3.38 - 3.06 (m, 6H), 2.90 (s, 3H), 2.87- 2.78 (m, 1H), 2.25 - 2.11 (m, 2H), 1.87 - 1.56 (m, 4H), 1.08 (t, J= 7.2 Hz, 3H).

[0317] 13C NMR (126 MHz, CDCb) 6 187.7, 167.5, 162.3, 155.6 (4C), 151.3 (dd. Ja = 250.3 Hz, Jc-CF= 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, Jc-CF= 11.3 Hz, 2C), 136.8 (2C), 133.8 (2C), 130.9 (dd, JC-C-CF = 6.3 Hz, Jc-C-CF = 3.7 Hz, 2C), 114.3 (dd, JC. CF= 20.1 Hz, JC. CF= 7.5 Hz, 4C), 98.4, 62.8, 53.5, 48.2, 44.0, 43.2, 33.5, 28.5, 26.9, 22.6, 13.7. One low-field carbon not observed.

[0318] 19F NMR (376 MHz, CDCb) 5 -133.2 (d, JFF= 20.2 Hz, 2F), -157.7 (t, JFF= 18.8 Hz, IF). HR-APCI m / z calcd for C33H33F6N6O2 [M+H] = 659.2563, found 659.2563.

[0319]

[0320] 7V-(3,5-bis( Zi)-3-Chloro-4,5-difluorobenzylidene)-4-oxocyclohexyl)-2-((2-(pyrrolidin-l-yl)ethyl)ami-no)pyrimidine-5-carboxamide (JCS181)

[0321] 'H NMR (500 MHz, CDCb) 58.69 (s, 2H), 7.66 (s, 2H), 7.26 - 7.14 (m, 5H), 6.63 (m, 1H), 4.51 > 4.44 (m, 1H), 3.67 - 3.58 (m, 2H), 3.24 (br d, J= 14.8 Hz, 2H), 3.05 (dd, J= 15.6, 8.4 Hz, 2H), 2.86 (t, J= 6.1 Hz, 2H), 2.72 (t, J= 6.4 Hz, 4H), 1.90- 1.80 (m, 4H).

[0322] 13C NMR (126 MHz, CDCb) 5 187.3, 164.2, 163.0, 157.6, 150.7 (dd, JCF = 253.2 Hz, JC-CF = 12.6 Hz, 2C), 150.3 (dd, JCF= 249.4 Hz, JC-CF = 12.6 Hz, 2C), 136.7 (2C), 133.9 (2C), 131.9 (dd, Jc.c. CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 127.4 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 123.2 (d, JC-CF = 17.6 Hz, 2C), 117.4 (d, JC-CF = 17.6 Hz, 2C), 116.5, 54.4, 54.1, 44.6, 39.8, 33.8, 23.6.

[0323] 19F NMR (376 MHz, CDCb) 5 -133.3 (d, JFF= 20.2 Hz, IF), -135.7 (d, JFF= 20.2 Hz, IF).

[0324] HR-APCI m, 'z calcd for C31H28CI2F4N5O2 [M+H] = 648.1150, found 648.1150.

[0325]

[0326] A-(3,5-bis((£')-3-Chloro-4,5-difluorobenzylidene)-4-oxocyclohexyl)-2-((((5)-l-ethylpyrrolidin-2-yl)methyl)amino)pyrimidine-5-carboxamide (JCS182)

[0327] 'H NMR (500 MHz, CDCb) 58.69 (s, 2H), 7.66 (s, 2H), 7.26 - 7.14 (m, 4H), 6.55 (, 1H), 6.42 (m, 1H), 4.51 - 4.44 (m, 1H), 3.72 - 3.62 (m, 2H), 3.42 - 3.32 (m, 1H), 3.22 (bd, J = 14.3 Hz, 2H), 3.07 (dd, J= 15.6, 8.1 Hz, 2H), 2.93 - 2.69 (m, 2H), 2.36 - 2.15 (m, 2H), 1.95 - 1.56 (m, 4H), 1.10 (t, J= 7.2Hz, 3H).

[0328] 13C NMR (126 MHz, CDCb) 5 187.3, 164.3, 163.4, 157.8 (d), 150.7 (dd, JCF= 253.2 Hz, Jc. CF = 12.6 Hz, 2C), 150.3 (dd, JCF= 249.4 Hz, JC-CF = 12.6 Hz, 2C), 136.7 (2C), 133.9 (2C), 131.9 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 127.4 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 123.2 (d, JC. CF= 17.6 Hz, 2C), 117.4 (d, JC-CF = 17.6 Hz, 2C), 116.3, 62.8, 53.6, 48.4, 44.5, 42.8, 33.7, 28.4, 22.7, 13.6.

[0329] 19F NMR (376 MHz, CDCb) 5 -133.2 (d, JFF= 20.2 Hz, IF), -135.6 (d, JFF= 20.2 Hz, IF).

[0330] HR-APCI m / z calcd for C32H30CI2F4N5O2 [M+H] = 662.1707, found 662.1713.

[0331]

[0332] AL(3,5-bis((£')-3-Chloro-4,5-difluorobenzylidene)-4-oxocyclohexyl)-4-methyl-2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxamide (JCS183)

[0333] 'H NMR (500 MHz, CDC13) 58.47 (s, 1H), 7.68 (s, 2H), 7.30 - 7.14 (m, 6H), 4.51 - 4.34 (m, 1H), 3.81 - 3.51 (m, 2H), 3.25 - 3.08 (m, 4H), 3.03 - 2.61 (m, 6H), 2.46 (s, 3H), 1.97 - 1.78 (m, 4H).

[0334] 13C NMR (126 MHZ, CDC13) 5187.3, 166.3, 161.7, 157.3 (2C), 150.7 (dd, JCF = 253.2 Hz, Jc. CF = 12.6 Hz, 2C), 150.3 (dd, JCF = 249.4 Hz, JC-CF = 12.6 Hz, 2C), 137.1, 136.1, 134.2 (2C), 131.8 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 127.3 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 123.0 (d, JC-CF = 17.6 Hz, 2C), 118.0, 117.3 (d, JC-CF = 17.6 Hz, 2C), 54.4, 54.1, 44.7, 39.3, 33.5, 23.4 (2C overlapped).

[0335] 19F NMR (376 MHz, CDCb) 5 -133.3 (d, JFF= 20.2 Hz, IF), -135.7 (d, JFF= 20.2 Hz, IF).

[0336] HR-APCI m / z calcd for C32H30CI2F4N5O2 [M+H] = 662.1707, found 662.1709.

[0337]

[0338] A-(3,5-bis((£')-3-Chloro-4,5-difluorobenzylidene)-4-oxocyclohexyl)-2-((((5)-l-ethylpyrrolidin-2-yl)methyl)amino)-4-methylpyrimidine-5-carboxamide (JCS184)

[0339] 'H NMR (500 MHz, CDCI3) 58.21 (s, 1H), 7.66 (s, 2H), 7.26 - 7.14 (m, 4H), 6.39 (m, 1H), 6.05 (m, 1H), 4.51 - 4.44 (m, 1H), 3.65 - 3.59 (m, 1H), 3.33 - 3.21 (m, 2H), 3.16 (bd, J= 14.3 Hz, 2H), 2.87 (dd, J= 15.6, 8.1 Hz, 2H), 2.76 - 2.60 (m, 2H), 2.38 (s, 3H), 2.36 - 2.15 (m, 2H), 1.95 - 1.56 (m, 4H), 1.10 (t, J= 7.2 Hz, 3H).13C NMR (126 MHz, CDC13) 5 187.2, 166.7, 162.2, 156.7, 150.7 (dd, JCF = 253.2 Hz, JC. CF = 12.6 Hz, 2C), 150.3 (dd, JCF = 249.4 Hz, JC. CF = 12.6 Hz, 2C), 136.7, 133.9, 131.9 (dd, JC-c-cF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 127.4 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 123.2 (d, JC-CF = 17.6 Hz, 2C), 117.5, 117.4 (d, JC-cF= 17.6 Hz, 2C), 62.9, 53.5, 48.3, 44.3, 42.6, 33.1, 28.3, 23.2, 22.7, 13.4. One low-field carbon not observed.

[0340] 19F NMR (376 MHz, CDCI3) 5 -133.2 (d, JFF= 20.2 Hz, IF), -135.6 (d, JFF= 20.2 Hz, IF).

[0341] HR-APCI m / z calcd for C33H32CI2F4N5O2 [M+H] = 676.1863, found 676.1868.

[0342]

[0343] 7V-(4-Oxo-3,5-bis((E)-2,3,4,5-tetrafluorobenzylidene)cyclohexyl)-2-((2-(pyrrolidin-l-yl)ethyl)amino)-pyrimidine-5-carboxamide (JCS185)

[0344] 'H NMR (500 MHz, CDCI3) 58.29 (s, 1H), 7.72 (s, 2H), 7.00 - 6.95 (m, 3H), 6.12 (m, 1H), 4.51 > 4.44 (m, 1H), 3.62 - 3.57 (m, 2H), 3.10 (br d, J= 14.8 Hz, 2H), 2.94 (dd, J= 15.6, 8.4 Hz, 2H), 2.85 (t, J= 6.1 Hz, 2H), 2.74 (t, J= 6.4 Hz, 4H), 1.87- 1.77 (m, 4H).

[0345] 13C NMR (126 MHz, CDCI3) 6 186.4, 164.1, 162.9, 158.6 (2C), 157.4 (2C), 146.8 (dd, JCF = 250.3 Hz, JG-CF = 11.3 Hz, 2C), 145.9 (dd, JCF = 250.3 Hz, JG-CF = 11.3 Hz, 2C), 136.0 (2C), 129.8 (2C), 119.2 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 116.3, 111.4 (d, JC-CF = 20.1 Hz, 4C), 54.4, 54.0, 44.4, 39.6, 33.8, 23.4. One low-field carbon not observed.

[0346] 19F NMR (376 MHz, CDCb) 5 -136.4 - -136.5 (m, IF), -138.3 (dd, JFF= 22.5, 11.3 Hz, IF), -152.7 (t, JFF= 18.8 Hz, IF). -154.4 (t, JFF= 18.8 Hz, IF).

[0347] HR-APCI m / z calcd for C31H26F8N5O2 [M+H] = 652.1953, found 652.1961.

[0348]

[0349] 4-Methyl-7V-(4-oxo-3,5-bis( E')-2,3,4,5-tetrafluorobenzylidene)cyclohexyl)-2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxamide (JCS186)

[0350] 'H NMR (500 MHz, CDC13) 58.61 (s, 1H), 7.72 (s, 2H), 7.00 - 6.95 (m, 3H), 6.73 (m, 1H), 4.44 > 4.38 (m, 1H), 3.62 - 3.57 (m, 2H), 3.04 (br d, J= 14.8 Hz, 2H), 2.74 (dd, J= 15.6, 8.4 Hz, 2H), 2.71 (t, J= 6.1 Hz, 2H), 2.63 (t, J= 6.4 Hz, 4H), 2.38 (s, 3H), 1.87 - 1.77 (m, 4H).

[0351] 13C NMR (126 MHz, CDCI3) 6 186.4, 166.6, 161.8, 156.8 (4C), 146.8 (dd, JCF= 250.3 Hz, Jc-CF= 11.3 Hz, 2C), 145.9 (dd, JCF = 250.3 Hz, Jc-CF= 11.3 Hz, 2C), 135.8 (2C), 130.4 (2C), 119.2 (dd, JC-C-CF = 6.3 Hz, Jc-C-CF = 3.7 Hz, 2C), 117.5, 111.4 (d, JC-CF = 20.1 Hz, 4C), 54.5, 53.9, 44.4, 39.4, 33.5, 23.5, 23.4. Several low-field carbons not observed.

[0352] 19F NMR (376 MHz, CDCb) 5 -136.5 - -136.6 (m, IF), -138.3 (dd, JFF = 22.5, 11.3 Hz, IF), -152.7 (t, JFF= 18.8 Hz, IF). -154.5 (t, JFF= 18.8 Hz, IF).

[0353] HR-APCI m / z calcd for C32H28F8N5O2 [M+H] = 666.2109, found 666.2111.

[0354]

[0355] 2-((((. S)-l-Etliylpyrrolidiii-2-yl)inetliyl)aniino)-\-(4-oxo-3.5-bis(( / :)-2.3.4.5-tetrafluorobenzylidene)-cyclohexyl)pyrimidine-5-carboxamide (JCS187)

[0356] 'H NMR (500 MHz, CDCI3) 6 8.67 (s, 2H), 7.71 (s, 2H), 7.07 - 7.04 (m, 3H), 6.60 (m, 1H), 4.50 - 4.46 (m, 1H), 3.75 - 3.68 (m, 1H), 3.50 - 3.25 (m, 2H), 3.05 (br d, J= 15.9 Hz, 2H), 3.01 - 2.86 (m, 4H), 2.45 - 2.29 (m, 2H), 2.01 - 1.61 (m, 4H), 1.14 (t, J= 7.2 Hz, 3H).13C NMR (126 MHz, CDC13) 6 186.4, 164.1, 163.3, 158.1 (2C), 157.4 (2C), 146.8 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 145.9 (dd, JC = 250.3 Hz, JC. CF = 11.3 Hz, 2C), 136.0 (2C), 129.8 (2C), 119.2 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 116.3, 111.4 (d, JC-CF = 20.1 Hz, 4C), 63.3, 53.5, 48.7, 44.4, 42.6, 33.9, 28.3, 22.5, 12.8. One low-field carbon not observed.

[0357] 19F NMR (376 MHz, CDC13) 5 -136.4 - -136.5 (m, IF), -138.3 (dd, JFF = 22.5, 11.3 Hz, IF), -152.7 (t, JFF = 18.8 Hz, IF). -154.4 (t, JFF = 18.8 Hz, IF).

[0358] HR-APCI m / z calcd for C32H28F8N5O2 [M+H] = 666.2109, found 666.2111.

[0359]

[0360] 2-((((5)-l-Ethylpyrrolidin-2-yl)methyl)amino)-4-methyl-A-(4-oxo-3,5-bis((£)-2,3,4,5-tetra-fluoro-benzylidene)cyclohexyl)pyrimidine-5-carboxamide (JCS188)

[0361] 'H NMR (500 MHz, CDC13) 6 8.26 (s, 1H), 7.75 (s, 2H), 7.07 - 6.88 (m, 3H), 6.13 (m, 1H), 4.50 - 4.46 (m, 1H), 3.75 - 3.68 (m, 1H), 3.42 - 3.26 (m, 2H), 3.05 (br d, J= 15.9 Hz, 2H), 3.01 -2.86 (m, 4H), 2.40 (s, 3H), 2.35 - 2.24 (m, 2H), 1.97 - 1.64 (m, 4H), 1.12 (t, J = 7.2 Hz, 3H).

[0362] 13C NMR (126 MHz, CDC13) 6 186.4, 166.6, 162.1, 156.9 (4C), 146.8 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 145.9 (dd, JCF = 250.3 Hz, Jc-CF= 11.3 Hz, 2C), 136.0 (2C), 129.8 (2C), 119.2 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 117.5, 111.4 (d, JC. CF = 20.1 Hz, 4C), 62.9, 53.5, 48.5, 44.50, 42.5, 33.6, 28.3, 22.7, 22.6, 22.6. Several low-field carbons not observed.

[0363] 19F NMR (376 MHz, CDC13) 5 -136.4 - -136.5 (m, IF), -138.3 (dd, JFF = 22.5, 18.8 Hz, IF), -152.7 (t, JFF = 18.8 Hz, IF). -154.5 (t, JFF= 18.8 Hz, IF).

[0364] HR-APCI m / z calcd for C33H3oF8N502 [M+H] = 680.2266, found 680.2273.

[0365]

[0366] 2-((2-(lH-imidazol-l-yl)ethyl)amino)-A-(4-oxo-3,5-bis((£)-3,4,5-trifluorobenzylidene)cyclohexyl)pyrimidine-5-carboxamide (JCS189)

[0367] 'H NMR (500 MHz, Methanol-< / 4) 6 8.65 (s, 2H), 7.92 (s, 1H), 7.63 (s, 2H), 7.30 -7.23 (m, 5H), 7.27 (s, 1H), 7.05 (s, 1H), 4.89 - 4.84 (m, 1H), 4.29 (t, J= 5.9 Hz, 2H), 3.79 (t, J= 5.9 Hz, 2H), 3.28 (br d, J= 14.8 Hz, 2H), 3.01 (dd, J= 15.6, 8.4 Hz, 2H).

[0368] 13C NMR (126 MHz, Methanol-< / 4) 6 187.3, 165.3, 162.9, 157.8 (4C), 151.3 (dd,. / ci = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 136.8, 135.2 (2C), 134.9 (2C), 131.8 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 125.4, 120.3, 116.8, 114.2 (d, JC-CF = 20.1 Hz, 4C), 54.42, 46.51, 45.32, 32.84.

[0369] 19F NMR (376 MHz, Methanol-< / 4) 6 -136.5 (d, JFF = 20.2 Hz, 2F), -162.0 (t, JFF = 18.8 Hz, IF)

[0370] HR-APCI m / z calcd for C30H23F6N6O2 [M+H] = 613.1781, found 613.1781.

[0371]

[0372] 2-((2-(lH-pyrazol-l-yl)ethyl)amino)-7V-(4-oxo-3,5-bis((E)-3,4,5-trifluorobenzylidene)cyclohexyl)pyrimidine-5-carboxamide (JCS190) 'H NMR (500 MHz, Methanol-< / 4) 6 8.64 (s, 2H), 7.94 (s, 1H), 7.61 (s, 2H), 7.27 -7.23 (m, 5H), 7.05 (s, 1H), 7.05 (s, 1H), 4.90 - 4.80 (m, 1H), 4.28 (t, J= 5.9 Hz, 2H), 3.78 (t, J= 5.9 Hz, 2H), 3.23 (br d, J= 14.8 Hz, 2H), 3.00 (dd, J= 15.6, 8.4 Hz, 2H).

[0373] 13C NMR (126 MHz, Methanol-< / 4) 6 187.3, 165.2, 162.9, 157.8 (4C), 151.3 (dd,. / ci = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 136.7, 135.2 (2C), 134.9 (2C), 131.8 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 125.6, 120.3, 116.8, 114.2 (d, JC-CF = 20.1 Hz, 4C), 54.37, 46.53, 45.28, 32.80.

[0374] 19F NMR (376 MHz, Methanol-< / 4) 6 -136.5 (d, JFF = 20.2 Hz, 2F), -162.0 (t, JFF = 18.8 Hz, IF)

[0375] HR-APCI m / z calcd for C30H23F6N6O2 [M+H] = 613.1781, found 613.1781.

[0376]

[0377] 2-((2-(lH-imidazol-l-yl)ethyl)amino)-N-(3,5-bis((Zi)-3,4-difluoro-5-methoxybenzylidene)-4-oxocyclohexyl)pyrimidine-5-carboxamide (JCS191)

[0378] 'H NMR (500 MHz, CDCh) 6 8.73 (s, 2H), 7.73 (s, 2H), 7.49 (s, 1H), 7.35 (s, 1H), 6.90 - 6.77 (m, 5H), 6.67 (s, 1H), 6.22 (s, 1H), 4.50 - 4.33 (m, 1H), 4.34 (t, J= 6.1 Hz, 2H), 3.95 - 3.89 (m, 8H), 3.26 (br d, J= 14.8 Hz, 2H), 3.10 (dd, J= 15.6, 8.4 Hz, 2H).

[0379] 13C NMR (126 MHz, CDCh) 6 187.6, 163.4, 161.5 (3C), 150.9 (dd, JC-F = 248.2, 11.3 Hz, 2C), 149.1 (d, JC-F = 8.8, 5.0 Hz, 2C), 141.4 (dd,. / c-i = 246.9, 13.8 Hz, 2C), 139.8, 138.3 (2C), 132.7 (2C), 130.6 (dd, JC-F = 8.8, 5.0 Hz, 2C), 130.0, 117.0, 111.2 (2C), 110.5 (d, JC-F = 18.9 Hz, 2C), 105.7, 56.8, 50.6, 44.7, 41.7, 33.6.

[0380] 19F NMR (376 MHz, CDCh) 6 -136.2 (d, JFF = 18.8 Hz, IF), -156.5 (d, JFF = 18.8 Hz, IF).

[0381] HR-APCI m / z calcd for C32H39F4N6O4 [M+H] = 637.2180, found 637.2180.

[0382]

[0383] 2-((2-(lH-pyrazol-l-yl)ethyl)amino)-N-(3,5-bis((E)-3,4-difluoro-5-methoxybenzylidene)-4-oxocyclohexyl)pyrimidine-5-carboxamide (JCS192)

[0384] 'H NMR (500 MHz, CDCh) 6 8.64 (s, 2H), 7.73 (s, 2H), 7.48 (s, 1H), 7.32 (s, 1H), 6.86 - 6.79 (m, 5H), 6.45 (s, 1H), 6.40 (s, 1H), 4.51 - 4.48 (m, 1H), 4.32 (t, J= 6.1 Hz, 2H), 3.95 - 3.89 (m, 8H), 3.24 (br d, J= 14.8 Hz, 2H), 3.10 (dd, J= 15.6, 8.4 Hz, 2H).

[0385] 13C NMR (126 MHz, CDCh) 6 187.6, 163.8, 162.3 (3C), 150.9 (dd, JC-F = 248.2, 11.3 Hz, 2C), 149.2 (d,. / c-i = 8.8, 5.0 Hz, 2C), 141.4 (dd,. / c-i = 246.9, 13.8 Hz, 2C), 139.8, 138.4 (2C), 132.6 (2C), 130.6 (dd, JC-F = 8.8, 5.0 Hz, 2C), 129.9, 117.0, 111.2 (2C), 110.5 (d, JC-F = 18.9 Hz, 2C), 105.7, 56.8, 50.6, 44.7, 41.7, 33.5.

[0386] 19F NMR (376 MHz, CDCh) 6 -136.2 (d, JFF = 18.8 Hz, IF), -156.4 (d, JFF = 18.8 Hz, IF).

[0387] HR-APCI m / z calcd for C32H39F4N6O4 [M+H] = 637.2180, found 637.2180.

[0388]

[0389] 2-((((5)-l-ethylpyrrolidin-2-yl)methyl)amino)-A-methyl-A-(4-oxo-3,5-bis((£')-3,4,5-trifluorobenzylidene)cyclohexyl)pyrimidine-5-carboxamide (JCS193)

[0390] 'H NMR (500 MHz, CDCh) 68.37 (s, 2H), 7.64 (s, 2H), 7.10 - 6.96 (m, 4H), 6.09 (s, 1H), 4.57 - 4.45 (m, 1H), 3.63 - 3.59 (m, 1H), 3.32 - 3.27 (m, 1H), 3.19 - 3.14 (m, 1H), 3.12 (s, 3H), 3.10 - 3.04 (m, 4H), 2.87 - 2.79 (m, 1H), 2.69 - 2.61 (m, 1H), 2.26 - 2.15 (m, 2H), 1.93 - 1.54 (m, 4H), 1.09 (t, J= 7.2 Hz, 3H).

[0391] 13C NMR (126 MHz, CDCh) 6 186.8, 168.3, 162.6, 158.1 (4C), 151.3 (dd, JCF = 250.3 Hz, Jc-CF = 11.3 Hz, 2C), 151.2 (dd,. / ci = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 136.2 (2C), 134.4 (2C), 131.0 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 117.8, 114.2 (dd, JC-CF = 13.8 Hz, Jc-CF = 3.7 Hz, 4C), 62.3, 53.4, 51.3, 48.0, 42.7, 31.0 (2C overlapped), 28.3, 22.7, 13.8.

[0392] 19F NMR (376 MHz, CDCh) 6 -133.1 (d, JFF = 22.5 Hz, 2F), -157.7 (t,. / IT = 18.8 Hz, IF).

[0393] HR-APCI m / z calcd for C33H32F6N5O2 [M+H] = 644.2454, found 644.2462.

[0394]

[0395] A-methyl-2-((((5)-l-methylpyrrolidin-2-yl)methyl)amino)-A-(4-oxo-3,5-bis((£)-3,4,5-trifluorobenzylidene)cyclohexyl)pyrimidine-5-carboxamide (JCS194)

[0396] 'H NMR (500 MHz, CDCh) 68.38 (s, 2H), 7.65 (s, 2H), 7.07 - 7.00 (m, 4H), 6.06 (s, 1H), 4.56 - 4.44 (m, 1H), 3.70 - 3.61 (m, 1H), 3.36 - 3.29 (m, 1H), 3.12 (s, 3H), 3.10 - 3.04 (m, 4H), 2.51 - 2.44 (m, 1H), 2.32 (s, 3H), 2.27 - 2.21 (m, 1H), 1.93 - 1.83 (m, 1H), 1.77 -1.61 (m, 4H).

[0397] 13C NMR (126 MHz, CDCh) 6 186.8, 168.3, 162.7, 158.1 (4C), 151.3 (dd, JCF = 250.3 Hz, Jc-CF = 11.3 Hz, 2C), 151.2 (dd,. / ci = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 136.3 (2C), 134.4 (2C), 131.0 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 117.9, 114.3 (dd, JC-CF = 13.8 Hz, Jc-CF = 3.7 Hz, 4C), 63.9, 57.2, 51.3, 42.2, 40.3, 31.0 (2C overlapped), 28.3, 22.6.

[0398] 19F NMR (376 MHz, CDCh) 6 -133.0 (d, JFF = 22.3 Hz, 2F), -157.6 (t, JFF = 18.8 Hz, IF).

[0399] HR-APCI m / z calcd for C32H30F6N5O2 [M+H] = 630.2298, found 630.2303.

[0400]

[0401] 2-((2-(lH-imidazol-l-yl)ethyl)amino)-V-methyl-V-(4-oxo-3,5-bis((E')-3,4,5-trifluorobenzylidene)cyclohexyl)pyrimidine-5-carboxamide (JCS195)

[0402] 'H NMR (500 MHz, CDCh) 68.34 (s, 2H), 7.65 (s, 2H), 7.49 (s, 1H), 7.06 - 7.02 (m, 4H), 6.69 (s, 1H), 6.89 (s, 1H), 6.41 (s, 1H), 4.59 - 4.40 (m, 1H), 4.16 (t, J= 5.9 Hz, 2H), 3.77 (t, J= 5.9 Hz, 2H), 3.12 (s, 3H), 3.10 - 3.05 (m, 4H).

[0403] 13C NMR (126 MHz, CDCh) 6 186.8, 167.9, 162.0, 158.0 (4C), 151.3 (dd, JCF = 250.3 Hz, Jc-CF = 11.3 Hz, 2C), 151.2 (dd,. / ci = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 137.3, 137.1 (2C), 136.3 (2C), 131.0 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 129.0, 119.2, 119.0, 114.2 (d, Jc-CF = 20.1 Hz, 4C), 51.1, 46.3, 42.3, 32.6, 31.0.

[0404] 19F NMR (376 MHz, CDCh) 6 -133.0 (d, JFF = 20.2 Hz, 2F), -157.6 (t,. / IT = 18.8 Hz, IF)

[0405] HR-APCI m / z calcd for C31H25F6N6O2 [M+H] = 627.1937, found 627.1939.

[0406]

[0407] 2-((2-(lH-pyrazol-l-yl)ethyl)amino)-A-methyl-A-(4-oxo-3,5-bis((£')-3,4,5-trifluorobenzylidene)cyclohexyl)pyrimidine-5-carboxamide (JCS196) 'H NMR (500 MHz, CDCh) 6 8.36 (s, 2H), 7.65 (s, 2H), 7.46 (s, 1H), 7.31 (s, 1H), 7.06 - 7.02 (m, 4H), 6.23 (s, 1H), 6.18 (s, 1H), 4.59 - 4.40 (m, 1H), 4.31 (t, J= 5.9 Hz, 2H), 3.87 (t, J= 5.9 Hz, 2H), 3.11 (s, 3H), 3.10 - 3.05 (m, 4H).

[0408] 13C NMR (126 MHz, CDCh) 6 186.8, 168.0, 162.1, 158.0 (4C), 151.3 (dd, JCF = 250.3 Hz, Jc-CF = 11.3 Hz, 2C), 151.2 (dd,. / ci = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 139.9, 136.3 (2C), 134.3 (2C), 131.0 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 129.8, 118.7, 114.2 (d, JC-CF = 20.1 Hz, 4C), 105.5, 50.8, 41.6 (2C overlapped), 31.0 (2C overlapped).

[0409] 19F NMR (376 MHz, CDCh) 6 -133.0 (d, JFF = 20.2 Hz, 2F), -157.6 (t,. / IT = 18.8 Hz, IF)

[0410] HR-APCI m / z calcd for C31H25F6N6O2 [M+H] = 627.1940, found 627.1937

[0411]

[0412] N-(3,5-bis((£')-3,4-difluoro-5-methoxybenzylidene)-4-oxocyclohexyl)-4,6-dimethyl-2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxamide (JCS197)

[0413] 'H NMR (500 MHz, CDCh) 67.72 (s, 2H), 6.87 - 6.78 (m, 4H), 6.28 (s, 1H), 5.94 (s, 1H), 4.50 - 4.42 (m, 1H), 3.91 (s, 6H), 3.53 - 3.48 (m, 2H), 3.22 - 3.15 (m, 2H), 2.72 (t, J = 6.4 Hz, 4H), 2.62 (t, J= 6.1 Hz, 4H), 2.11 (s, 6H), 1.78 - 1.74 (m, 4H).

[0414] 13C NMR (126 MHz, CDCh) 6 187.4, 168.5, 164.0, 161.0 (2C), 150.9 (dd,. / c-i = 248.2, 11.3 Hz, 2C), 149.2 (dd, JC-F = 8.8, 5.0 Hz, 2C), 141.4 (dd, JC-F = 246.9, 13.8 Hz, 2C), 138.6 (2C), 132.4 (2C), 130.5 (dd, JC-F = 8.8, 5.0 Hz, 2C), 119.3, 111.0 (2C), 110.6 (d,. / c-i = 18.9 Hz, 2C), 56.7, 54.9, 53.9, 44.5, 39.5, 32.9, 23.4, 22.0.

[0415] 19F NMR (376 MHz, CDCh) 6 -136.2 (d, JFF = 18.8 Hz, IF), -156.4 (d, JFF = 18.8 Hz, IF).

[0416] HR-APCI m / z calcd for C35H38F4N5O4 [M+H] = 668.2854, found 668.2864.

[0417]

[0418] 7V-(3,5-bis((Zi)-3,4-difluoro-5-methoxybenzylidene)-4-oxocyclohexyl)-2-((((»y)-l-ethylpyrrolidin-2-yl)methyl)amino)-4,6-dimethylpyrimidine-5-carboxamide (JCS198) 'H NMR (500 MHz, CDCh) 67.74 (s, 2H), 7.88 - 7.78 (m, 4H), 6.16 (s, 1H), 5.81 (s, 1H), 4.51 - 4.47 (m, 1H), 3.91 (s, 6H), 3.63 - 3.56 (m, 1H), 3.33 - 3.16 (m, 6H), 2.93 - 2.85 (m, 1H), 2.77 - 2.70 (m, 1H), 2.36 - 2.22 (m, 2H), 2.13 (s, 6H), 1.93 - 1.60 (m, 4H), 1.11 (t, J = 7.2 Hz, 3H).

[0419] 13C NMR (126 MHz, CDCh) 6 187.4, 168.4, 164.1, 161.3 (2C), 150.9 (dd,. / c-i = 248.2, 11.3 Hz, 2C), 149.2 (dd, JC-F = 8.8, 5.0 Hz, 2C), 141.4 (dd, JC-F = 246.9, 13.8 Hz, 2C), 138.7 (2C), 132.3 (2C), 130.5 (dd,. / c-i = 8.8, 5.0 Hz, 2C), 119.5, 110.9 (2C), 110.6 (d,. / c-i = 18.9 Hz, 2C), 63.3, 56.7, 53.5, 48.5, 44.5, 42.5, 32.9, 28.2, 22.0, 13.3.

[0420] 19F NMR (376 MHz, CDCh) 6 -136.1 (d, JFF = 18.8 Hz, IF), -156.4 (d, JFF = 18.8 Hz, IF).

[0421] HR-APCI m / z calcd for C36H40F4N5O4 [M+H] = 682.3024, found 682.3010.

[0422]

[0423] 2-((((5)-l-ethylpyrrolidin-2-yl)methyl)amino)-4,6-dimethyl-A-(4-oxo-3,5-bis((£')- 3,4,5-trifluorobenzylidene)cyclohexyl)pyrimidine-5-carboxamide (JCS199) 'H NMR (500 MHz, CDCh) 67.70 (s, 2H), 7.07 - 7.04 (m, 4H), 6.12 (s, 1H), 5.75 (s, 1H), 4.53 - 4.47 (m, 1H), 3.63 - 3.56 (m, 1H), 3.30 - 3.17 (m, 6H), 2.91 -2.82 (m, 1H), 2.73 -2.66 (m, 1H), 2.32 -2.18 (m, 2H), 2.14 (s, 6H), 1.92- 1.58 (m, 4H), 1.11 (t, J= 7.2Hz, 3H).

[0424] 13C NMR (126 MHz, CDCh) 6 187.0, 168.6, 164.1, 161.3 (4C), 151.3 (dd, JCF = 250.3 Hz, Jc-CF = 11.3 Hz, 2C), 151.2 (dd,. / ci = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 137.4 (2C), 133.1 (2C), 130.9 (dd, JC-C-CF = 6.3 Hz, JC-C-CF = 3.7 Hz, 2C), 119.2, 114.3 (dd, JC-CF = 20.1 Hz, Jc-CF = 7.5 Hz, 4C), 63.0, 53.5, 48.3, 44.3, 42.5, 32.8, 28.3, 22.6, 22.0, 13.4.

[0425] 19F NMR (376 MHz, CDCh) 6 -133.1 (d, JFF = 20.2 Hz, 2 F), -156.3 (t, JFF = 18.8 Hz, IF).

[0426] HR-APCI m / z calcd for C34H34F6N5O2 [M+H] = 658.2627, found 658.2689.

[0427]

[0428] 4,6-dimethyl-7V-(4-oxo-3,5-bis((E)-3,4,5-trifluorobenzylidene)cyclohexyl)-2-((2-(pyrrolidin-l-yl)ethyl)amino)pyrimidine-5-carboxamide (JCS200)

[0429] 'H NMR (500 MHz, CDCI3) 57.66 (s, 2H), 7.06 - 7.03 (m, 4H), 6.85 (s, 1H), 6.05 (s, 1H), 4.46 - 4.42 (m, 1H), 3.55 - 3.51 (m, 2H), 3.17 - 3.16 (m, 4H), 2.84 (t, J= 6.1 Hz, 2H), 2.78 (t, J= 6.4 Hz, 4H), 2.09 (s, 6H), 1.84 - 1.83 (m, 4H).

[0430] 13C NMR (126 MHz, CDCI3) 6 187.2, 168.7, 164.1, 160.9 (4C), 151.3 (dd. Ja = 250.3 Hz, Jc-CF= 11.3 Hz, 2C), 151.2 (dd, JCF = 250.3 Hz, JC-CF = 11.3 Hz, 2C), 137.1 (2C), 133.5 (2C), 131.0 (dd, Jc-C-CF = 6.3 Hz, Jc-C-CF = 3.7 Hz, 2C), 119.6, 114.4 (dd, JC-CF = 20.1 Hz, JC-CF = 7.5 Hz, 4C), 55.0, 54.0, 44.5, 39.21, 33.0, 23.4, 22.0.

[0431] 19F NMR (376 MHz, CDCb) 5 -133.1 (d, JFF = 20.2 Hz, 2 F), -157.5 (t, JFF = 18.8 Hz, IF).

[0432] HR-APCI m / z calcd for C33H32F6N5O2 [M+H] = 644.2449, found 644.2454. Compounds of the present disclosure

[0433]

[0434]

[0435]

[0436] Example 2: Biological Activity of Compounds of this Disclosure

[0437] Cell killing potency

[0438] 27 analogs of JC099 were developed, including JCS162-188, and were screened as potential candidates by combining cell killing, PARP1 trapping, and ATR inhibition potency. Cell killing potency was detected by a CCK8 assay in prostate and breast cancer cells. As shown in FIG. 1A-D, JC SI 64 / 165 / 167 / 168 / 169 / 173 had higher cell killing potency than JC099 in PC3 and SUM149PT-BRCAlmutcells, while JCS 166 / 170 / 171 / 172 exhibited similar cell killing potency relative to JC099. The activities of JCS164 and JCS165 were higher than JCS141 and JCS152. Some other analogs, including JCS162 / 163, also had moderate potency of cell killing, whereas less than JC099. JCS174-188 are analogs for JCS164, so JCS164 was used for comparison. As shown in FIG. 1E-H and FIG. 1J, JCS175 / 178 had higher cell killing potency than JCS164 in PC3 and SUM149PT-BRCAlmut cells, while JCS174 / 177 exhibited similar cell killing potency relative to JCS164. Some other analogs, including JCS176 / 179-192, also had moderate potency of cell killing, whereas less than JCS 164. As shown in FIG. II, the activities of JCS164 / 165 / 169 were higher than JCS136, which is a high potential candidate on our previous patent. In addition, new compounds had high cell killing potency in ovarian cancer and myeloma cells. As shown in FIG. 2A, cell viabilities were significantly reduced after the treatment with JCS 164 / 165 / 169 / 178 in cisplatin-resistant OVCAR / CR cells, and their potencies were higher than JCS136; JCS136 and JCS164 also decreased cell viabilities in a dose-dependent manner in both cisplatin-sensitive and -resistant SKOV3 and SKOV3 / CR cells, and the potency of JCS164 was a little better than JCS136. As shown in FIG. 3, JCS 164 and JCS 165 significantly reduced cell viabilities in a dose-dependent manner in myeloma RPMI 8226 and MM. IS cells. As shown in Fig. 10A& B, JCS 193 / 194 / 199 showed higher cell killing potency than JCS164 in PC3 and SUM149PT-BRCAlmut cells, while JCS195 / 197 / 198 / 200 exhibited similar cell killing potency relative to JCS164. ATR inhibition

[0439] ATR inhibition potency of JC099 analogs was evaluated by decreasing OLA or UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites. As shown in FIG. 4, the phosphorylation of Rad 17 and CHK1 decreased more dramatically after the treatment with JCS161 / 164 / 165 / 167 / 168 / 169 than JC099, indicating greater ATR inhibition potential of JCS161 / 164 / 165 / 167 / 168 / 169 than JC099. The activities of JCS164 and JCS165 were higher than JCS 152 (FIG.4B), which is a high potential candidate on our previous patent. Some other analogs, including JCS166 / 170 / 171 also decreased OLA induced phosphorylation of Radl7 and CHK1, while their potencies were close to JC099. Compared to JC099, JCS 162 / 163 / 172 / 173 had less ATR inhibition potencies, albeit they also reduced OLA or UV induced phosphorylation ofRadl7 and CHK1. Additionally, the potency of JCS 178 in reducing the phosphorylation of Radl7 and CHK1 was higher than JCS164, indicating greater ATR inhibition potential than JCS164. JCS 174 / 175 / 181 / 182 / 184 has similar potency in ATR inhibition relative to JCS164, while JCS 176 / 177 / 179 / 180 / 183 / 185- 192 has less potency in ATR inhibition than JCS 164. In addition to prostate cancer cells, new compounds also exhibited activities in ATR inhibition in ovarian cancer and myeloma cells. As shown in FIG.

[0440] 5A, OLA induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites was dramatically reduced by JCS164 / 165 / 169 in OVCAR / CR cells, and their activities were higher than JCS136. Likely, UV induced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 sites was also significantly reduced by JCS136 and JCS164 in both cisplatin-sensitive SKOV3 (FIG. 5B) and A2780 (FIG. 5C) and cisplatin-resistant CP70 cells (FIG. 5D), while JCS164 exhibited higher potency than JCS136 in all the three cell lines. JCS164 and JCS165 also exhibited high potency in ATR inhibition in myeloma, evidenced by reduced phosphorylation of Radl7 at Ser 645 and CHK1 at Ser 345 after UV treatment in both RPMI 8226 and MM.1 S cells (FIG. 6). As shown in Fig. 11 A& B, OLA or UV induced phosphorylation of Radl7 and CHK1 decreased more dramatically after the treatment with JCS193 / 194 than JCS164, indicating greater ATR inhibition potential of JCS 193 / 194 than JCS 164, while JCS 195 showed close potency to JCS164. As shown in Fig. 11C& D, JCS199 reduced OLA or UV induced phosphorylation of Radl7 and CHK1 decreased more dramatically than JCS164, indicating greater ATR inhibition potential of JCS199 than JCS164, while JCS197 / 198 showed similar potency to JCS 164. As shown in Fig. 1 IE, the activity of JCS200 in decreasing phosphorylation of Radl7 and CHK1 is close to JCS164, indicating similar activity. PARP1 trapping potency

[0441] The chromatin-bound proteins were isolated from cells with treatment of JC099 or its analogs to analyze the PARP1 trapped on DNA lesions. As shown in FIG. 7, an increase of PARP1 in the chromatin-bound fraction were observed in PC-3 cells with treatment of JC SI 64 / 165 / 166 / 167 / 172 / 173 relative to JC099, indicating higher potency in PARP1 -trapping of JCS164 / 165 / 166 / 167 / 172 / 173 than JC099. JCS161 / 162 / 163 / 168 / 169 / 170 / 171 has similar potency in PARP1 trapping compared to JC099. Compared with JCS164, JCS 177 / 180 / 181 / 182 / 183 / 185 / 186 / 187 / 188 has higher potency in PARP1 trapping, while JCS174 / 175 / 176 / 178 / 179 / 184 has similar potency in PARP1 trapping, evidenced by the increased accumulation of PARP1 in the chromatin-bound fraction. As shown in Fig. 12, an increase of PARP1 in the chromatin-bound fraction were observed in PC-3 cells with treatment of JCS190 / 191 / 198 / 199 relative to JCS164, indicating higher PARP1 -trapping potency of JCS190 / 191 / 198 / 199 than JCS164. JCS192-197 and JCS200 showed similar potency in PARP1 trapping compared to JCS 164, evidenced by the accumulation of PARP1 in the chromatin-bound fraction.

[0442] In vivo activity

[0443] To evaluate the anti -turn or activity of new compounds in vivo, three xenografts were used, including a Olaparib-resistant breast cancer (ORBC) PDX (Patient Derived Xenografts), a castration-resistant prostate cancer (CRPC) PDX, and cisplatin-resistant ovarian cancer xenografts (CDX). To generate ORBC and CRPC PDX, tumor tissue or cells were subcutaneously implanted in CB-17 / SCID mice. When the tumors reached -100 mm3, mice were administered orally (i.g.) new compounds at dose of 60 mg / kg five times weekly. Equivalent solvent was used as control. The longest (L) and shortest (W) tumor axes were measured, and tumor volume (mm3) was calculated as L * W2 / 2. In addition, the body weights of tumor-bearing mice were recorded twice a week to determine toxicity.

[0444] To generate CDX, cisplatin-resistant OVCAR5 / CR cells were subcutaneously implanted in Nude mice. When the tumors reached -80 mm3, mice were grouped randomly and administered orally (i.g.) with new compounds at dose of 60 mg / kg daily. Equivalent solvent was used as control. The longest (L) and shortest (W) tumor axes were measured, and tumor volume (mm3) was calculated as L x W2 / 2. In addition, the body weights of tumorbearing mice were recorded twice a week to evaluate toxicity. FIG. 8 shows the results obtained in ORBC PDX, which is resistant to PARP inhibitors. Compared to the vehicle controls, treatments with JC099 or JCS 144 / 152 / 164 / 165 suppressed tumor growth (FIG. 8B) and decreased tumor weight by -46-70% (FIG. 8A). At a dose of 60 mg / kg, JCS 164 had the best activity compared to all other compounds at the same dose level. And the effect of JCS164 at 60 mg / kg was close to that of JC099 at the higher dose of 100 mg / kg. JCS 165 decreased 55% tumor weight, also indicating high activity in tumor inhibition. Both JCS164 and JCS165 exhibited higher activities compared to JCS144 and JCS152, two high potential candidates on our previous patent. None of the compounds influenced body weights of tumor-bearing mice (FIG. 8C), indicating no obvious toxicity was observed.

[0445] FIG. 9 shows the results obtained in the CRPC PDX, which has acquired resistance to Enzalutamide. Compared to the vehicle controls, tumor growth was significantly suppressed by the treatment of JC099 or JCS164 / 165 / 169 (FIG.9B). JCS165 and JCS169 decreased tumor weights by 56.9% and 63.3%, respectively, which is higher than JC099 (FIG. 9A), indicating higher activities than JC099. The activity of JCS164 is close to JC099 in this model. None of the analogs reduced body weights of tumor-bearing mice (FIG. 9C), indicating no obvious toxicity was observed.

[0446] FIG. 13 shows the results that compare JCS169 / 178 / 194 with JCS164 and Olaparib. Compared to the vehicle controls and Olaparib, treatments with JCS164 or JCS169 / 178 / 194 dramatically suppressed tumor growth (FIG. 13B) and decreased tumor weight by -40-75% (FIG. 13A). JCS169, 178 and 194 all exhibited higher activities in tumor growth inhibition relative to JCS164. Among them, JCS178 showed the best potency by decreasing 73.9% tumor weight. None of the compounds significantly reduced body weights of tumor-bearing mice (FIG. 13C), indicating no obvious toxicity was observed.

[0447] In FIG. 14, cisplatin-resistant ovarian cancer CDX-bearing mice were treated with JCS 169 / 178 / 193 / 199 at dose of 60 mg / kg. Olaparib was treated for comparison. As shown in FIG. 14B, JCS169 / 178 / 193 / 199 all dramatically delayed tumor growth, and their activities were higher than Olaparib. Among them, JCS169, 178, and 199 showed close potency in tumor inhibition, evidenced by reducing -75% tumor weights, respectively (FIG. 14A). None of the analogs reduced body weights of tumor-bearing mice (FIG. 14C), indicating no obvious toxicity was observed. Plasma half-life

[0448] The plasma half-life of the compounds of this disclosure can be measured by any means known to one of skill in the art, for example, quantitative tandem LC-MS. The plasma half-life is the time taken for the concentration of a compound to reach half of its maximum concentration (Cmax) in the plasma following administration.

[0449] Bioavailability

[0450] The bioavailability of the compounds of this disclosure can be measured by any means known to one of skill in the art, for example, by isotopic labeling experiments and / or by quantitative tandem LC-MS. The absolute bioavailability may be determined by measuring the ratio of the product of the area under the curve (AUC) of the route of administration (ROA) of interest and the intravenous dose, and the product of the AUC value of intravenous AUC xD-administration and the dose for the ROA of interest; i.e., FAh<;= 100 X - — — -.

[0451]

[0452] Blood-Brain Barrier permeability

[0453] The Blood-Brain Barrier (BBB) permeability of the compounds of this disclosure can be measured by any means known to one of skill in the art, for example, by quantifying the degree of permeation across two confluent bilayers of the appropriate cells (e.g., brain microvascular endothelial cells, pericytes, and astrocytes) grown on both the upper and lower sides of a well of a transwell plate. The quantification may be performed, for example, by quantitative tandem LC-MS.

[0454] INCORPORATION BY REFERENCE

[0455] All publications and patents mentioned herein are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

[0456] EQUIVALENTS

[0457] While specific embodiments of the subject invention have been discussed, the above specification is illustrative and not restrictive. Many variations of the invention will become apparent to those skilled in the art upon review of this specification and the claims below. The full scope of the invention should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations.

Claims

We claim:

1. A compound having a structure represented by Formula I, or a pharmaceutically acceptable salt thereof:Iwherein:A is heterocyclyl or heteroaryl;ml is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;nl is 1, 2, 3, 4, or 5;X1is O, S, or NR10;X2is O, S, or NR11;X3is O, S, or NR12;each of R1, R2, R3, R4, R5, R6, R7and R8is independently selected from H, alkoxy (e.g., methoxy or ethoxy), aralkyloxy (e.g., benzyloxy), fluoro, chloro, bromo, and iodo; each of R10, R11, and R12is independently selected from H, alkyl, and aralkyl; and each R9is independently selected from H, halo, hydroxyl, amino, amido, alkyl, alkenyl, alkynyl, carboxyl, aryl, acetyl, ester, thioester, alkoxy, cyano, nitro, azido, alkylthio, cycloalkyl, heteroaryl, heterocyclylalkyl, and heterocyclyl.

2. The compound of claim 1, wherein the compound is not:

3. The compound of claim 1 or 2, wherein the compound has a structure represented by Formula la, or a pharmaceutically acceptable salt thereof:

4. The compound of any one of claims 1-3, wherein R1is alkoxy (e.g., methoxy or ethoxy).

5. The compound of any one of claims 1-3, wherein R1is aralkyloxy (e.g., benzyloxy).

6. The compound of any one of claims 1-3, wherein R1is fluoro.

7. The compound of any one of claims 1-6, wherein R2is fluoro.

8. The compound of any one of claims 1-7, wherein R3is alkoxy (e.g., methoxy or ethoxy).

9. The compound of any one of claims 1-7, wherein R3is fluoro.

10. The compound of any one of claims 1-7, wherein R3is chloro.

11. The compound of any one of claims 1-10, wherein R4is H.

12. The compound of any one of claims 1-10, wherein R4is fluoro.

13. The compound of any one of claims 1-12, wherein R5is H.

14. The compound of any one of claims 1-12, wherein R5is fluoro.

15. The compound of any one of claims 1-14, wherein R6is fluoro.

16. The compound of any one of claims 1-14, wherein R6is chloro.

17. The compound of any one of claims 1-16, wherein R7is fluoro.

18. The compound of any one of claims 1-17, wherein R8is fluoro.

19. The compound of any one of claims 1-17, wherein R8is alkoxy (e.g., methoxy or ethoxy).

20. The compound of any one of claims 1-17, wherein R8is aralkyloxy (e.g., benzyloxy).

21. The compound of any one of claims 1-20, wherein nl is 1.

22. The compound of any one of claims 1-20, wherein nl is 2.

23. The compound of any one of claims 1-22, wherein the compound has a structure represented by Formula lb, or a pharmaceutically acceptable salt thereof:

24. The compound of claim 23, wherein A is heterocyclyl e.g., pyrrolidine or N-alkylpyrrolidine).

25. The compound of claim 23, wherein A is heteroaryl (e.g., imidazolyl or pyrazolyl).

26. The compound of any one of claims 23-25, wherein ml is 1 or 2.

27. The compound of any one of claims 23-26, wherein A is selected fromindicates the point of attachment.

28. The compound of any one of claims 23-27, wherein R12is H.

29. The compound of any one of claims 23-27, wherein R12is alkyl (e.g., methyl).

30. The compound of any one of claims 23-28, wherein the compound has a structure represented by Formula Ic, or a pharmaceutically acceptable salt thereof:

31. The compound of any one of claims 23-28, wherein the compound has a structure represented by Formula Id, or a pharmaceutically acceptable salt thereof:Id.

32. The compound of any one of claims 23-28, wherein the compound has a structure represented by Formula le, or a pharmaceutically acceptable salt thereof:

33. The compound of any one of claims 23-28, wherein the compound has a structure represented by Formula If, or a pharmaceutically acceptable salt thereof:

34. The compound of any one of claims 23-28, wherein the compound has a structure represented by Formula Ig, or a pharmaceutically acceptable salt thereof:-n -35. The compound of any one of claims 23-28, wherein the compound has a structure represented by Formula Ih, or a pharmaceutically acceptable salt thereof:

36. The compound of any one of claims 23-28, wherein the compound has a structure represented by Formula li, or a pharmaceutically acceptable salt thereof:

37. The compound of claim 29, wherein the compound has a structure represented by Formula Ij, or a pharmaceutically acceptable salt thereof:

38. The compound of any one of claims 1-20 and 22, wherein the compound has a structure represented by Formula Ik, or a pharmaceutically acceptable salt thereof:

39. The compound of any one of claims 1-20 and 22, wherein the compound has a structure represented by Formula II, or a pharmaceutically acceptable salt thereof:

40. The compound of any one of claims 23-39, wherein R9is alkyl e.g., methyl or tri fluoromethyl).

41. The compound of any one of claims 23-39, wherein R9is amino (e.g., methylamino).

42. The compound of any one of claims 23-39, wherein R9is H.

43. The compound of claim 1, wherein the compound is selected from:pharmaceutically acceptable salt thereof.

44. The compound of claim 1, wherein the compound is selected from:acceptable salt thereof.

45. A pharmaceutical composition comprising the compound of any one of claims 1-44 and a pharmaceutically acceptable excipient.

46. A method of treating a cancer in a subject in need thereof, comprising administering to the subject an amount of a compound of any one of claims 1-44 or a pharmaceutically acceptable salt thereof.

47. The method of claim 46, wherein the cancer is breast cancer, head and neck cancer, lung cancer, prostate cancer, or ovarian cancer.

48. The method of claim 46, wherein the cancer is testicular cancer, cervical cancer, bladder cancer, esophageal cancer, mesothelioma, orbrain cancer (e.g., neuroblastoma).

49. The method of claim 47, wherein the cancer is castration-resistant.

50. The method of claim 48, wherein the cancer is castration-resistant prostate cancer (CRPC).

51. The method of any one of claims 46-50, wherein the cancer is relapsed.

52. The method of any one of claims 46-51, wherein the cancer is refractory.

53. The method of any one of claims 46-52, wherein the cancer is resistant to treatment with olaparib.

54. The method of any one of claims 46-53, wherein the cancer is resistant to treatment with cisplatin.

55. The method of any one of claims 46-54, wherein the cancer is resistant to treatment with enzalutamide.

56. A method of inhibiting repair of DNA in a subject in need thereof, comprising administering to the subject an amount of a compound of any one of claims 1-44 or a pharmaceutically acceptable salt thereof.

57. A method of inhibiting PARP and ATR in a subject in need thereof, comprising administering to the subject an amount of a compound of any one of claims 1-44 or a pharmaceutically acceptable salt thereof.

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