RU2805949C2 - Compounds for treatment of cancer - Google Patents

Abstract

FIELD: oncology.

SUBSTANCE: invention relates to a compound represented by the formula 15 or 18, or its pharmaceutically acceptable salt. The invention relates to a pharmaceutical composition having anticancer activity and containing an effective amount of a compound of formulas 15 and 18 or its pharmaceutically acceptable salt and at least one pharmaceutically acceptable carrier. The invention also relates to the use of the compound according to the invention to obtain a drug for the treatment, attenuation, reduction of severity, reduction of risk of developing or suppressing cancer and to obtain a drug for the treatment of a tumor or tumors with drug resistance.

EFFECT: technical result is compounds interacting with the binding site of cochicin, having anticancer activity.

14 cl, 12 dwg, 4 tbl, 7 ex

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Description

Declaration of State Interest

The present invention was made in whole or in part with government support from the Van Vleet Professorship (D.D.M.) and NIH grants R01CA148706, 1S10RR026337-01, 1S10OD010678-01 (W.L.). The US Government retains certain rights to this invention.

Field of technology

The present invention relates to compounds that interact with the binding site of colchicine having anticancer activity, to compositions containing these compounds, and to their use for the treatment of various forms of cancer.

State of the art

Cancer is the second leading cause of death in the United States, second only to heart disease. In the United States, cancer causes 1 in 4 deaths. The 5-year relative survival rate for all cancer patients diagnosed between 1996 and 2003 is 66%, compared with 50% between 1975 and 1977 (Cancer Facts & Figures American Cancer Society: Atlanta, GA (2008)). This improvement in survival reflects advances in early diagnosis and developments in treatment options. A major challenge in cancer research is to identify highly effective anticancer agents with low toxicity.

Prostate cancer is one of the most commonly diagnosed non-skin cancers in men in the United States and represents the second leading cause of cancer death, with 180,000 new cases and nearly 29,000 deaths expected this year. Patients with advanced prostate cancer are treated with androgen deprivation therapy (ADT), usually with luteinizing hormone-releasing hormone agonists (RLH) or bilateral orchiectomy. Antiandrogen therapy reduces not only testosterone levels, but also estrogen levels, since estrogen is produced by aromatization of testosterone, which is reduced by ADT. Estrogen deficiency caused by androgen deprivation therapy leads to significant side effects, including hot flashes, gynecomastia and mastalgia, bone loss, decreased bone quality and strength, osteoporosis and life-threatening fractures, irreversible lipid changes, increased risk of cardiovascular disease and myocardial infarction , depression and other mood changes.

Malignant melanoma is the most dangerous form of skin cancer and is responsible for approximately 75% of all skin cancer deaths. The incidence of melanoma is gradually increasing among the population of the Western world. Over the past 20 years, this number has doubled. Worldwide, approximately 160,000 new cases of melanoma are diagnosed annually, most often in men and Caucasians. According to a WHO report, approximately 48,000 melanoma-related deaths occur annually worldwide.

Currently, there are no effective treatments for metastatic melanoma. Metastatic melanoma is highly resistant to existing methods of chemotherapy, radiotherapy and immunotherapy. Metastatic melanoma has a very poor prognosis, with a median survival time of 6 months and a 5-year survival rate of less than 5%. Over the past 30 years, dacarbazine (DTIC) has been the only drug approved by the US Food and Drug Administration (FDA) for the treatment of metastatic melanoma. However, dacarbazine provides complete remission in only less than 5% of patients. In recent years, significant efforts have been made to combat metastatic melanoma. Combinations of DTIC with other chemotherapy drugs (eg, cisplatin, vinblastine, and carmustine) or the addition of interferon-α2b to DTIC did not provide improved survival compared with treatment with DTIC alone. More recently, clinical trials of antibodies and vaccines for the treatment of metastatic melanoma have also failed to achieve satisfactory efficacy. Ipilimumab (Yervoy) is a drug that uses a subject's immune system to fight melanoma. Ipilimumab is used to treat advanced melanoma that has spread beyond its original location. Targeted therapy uses drugs designed to target specific weak spots in cancer cells. Vemurafenib (Zelboraf) is a targeted therapy approved for the treatment of advanced melanoma that cannot be treated with surgery or melanoma that has spread throughout the body. Vemurafenib can only treat melanoma that has a specific genetic mutation.

Drugs that interact with tubulin/microtubules have been successfully used to treat a wide range of human cancers. They are often classified into two main categories: microtubule-stabilizing drugs (eg, taxanes, epothilones) and microtubule-destabilizing drugs (eg, vinca alkaloids, colchicine). Three main binding sites have been identified in α,β-tubulin subunits: binding sites for taxanes, vinca alkaloids, and colchicine. Although antimitotic agents that interact with the taxane or vinca alkaloid binding sites of tubulin are very effective in clinical oncology, there are currently no FDA-approved drugs that interact with the colchicine binding site. and US drugs for the treatment of cancer. Most colchicine-binding agents have high potency, a chemical structure that is relatively easy to optimize, selective toxicity to the tumor vasculature, and a promising ability to overcome multidrug resistance mediated by the P-glycoprotein (P-gp) efflux pump.

Some excellent agents for this approach are shown in Figure 1. Combretastatin A-4 (CA-4) is the most active member of the combretastatin family, isolated from the African tree Combretum caffrum. CA-4 has potent antitubulin activity through binding to the colchicine site and is the subject of phase II and III clinical trials. Replacing the olefin bridge in CA-4 with a carbonyl group results in phenstatin, which has activity and mechanism of action similar to CA-4. BPR0L075 and Oxi-6196 are 2-aroylindole and dihydronaphthalene analogs of CA-4 that provide potent inhibition of tubulin polymerization. Methylated chalcone SD400, which has an IC ₅₀ value against K562 human leukemia cells of 0.21 nM, is a highly active tubulin inhibitor. Podophyllotoxin is a toxic lignin of non-alkaloid nature and also has anticancer properties that can inhibit tubulin polymerization by binding to the colchicine binding site. Accordingly, compounds that interact with the binding site of colchicine have been of great interest to specialists in the field of medicinal chemistry in recent years.

Brief description of the invention

In one embodiment, the present invention provides a compound represented by Formula II:

(II)

Where

Q is S, NH or O;

Z represents CH or N;

A is substituted or unsubstituted phenyl; substituted or unsubstituted indolyl; or substituted or unsubstituted indazolyl;

wherein optional substituents at A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , - (CH ₂ ) _i NH ₂ , -(CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₁ independently represents hydrogen, O-alkyl, O-haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H, NHCO-alkyl, -O(CH ₂ ) _j OCH ₃ , -O(CH ₂ ) _j OH, - O(CH ₂ ) _j NHCH ₃ , -O(CH ₂ ) _j NH ₂ , -O-(CH ₂ ) _j N(CH ₃ ) ₂ , - OC(O)CF ₃ , -OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _j NH ₂ , -O(CH ₂ ) _j N-phthalimide or a combination thereof;

R ₂ represents hydrogen, O-alkyl, O-haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H, NHCO-alkyl, -O(CH ₂ ) _k OCH ₃ , -O(CH ₂ ) _k OH, - O(CH ₂ ) _k NHCH ₃ , -O(CH ₂ ) _k NH ₂ , -O-(CH ₂ ) _k N(CH ₃ ) ₂ , -OC (O)CF ₃ , -OC(O)CH ₂ Cl, - OCH ₂ Ph, -O(CH ₂ ) _k NH ₂ or -O(CH ₂ ) _k N-phthalimide;

i, j and k are independently integers from 0 to 5;

n is an integer from 1 to 4;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the present invention provides a compound represented by Formula XII:

Where

Q is S, NH or O;

X represents a bond or NH;

A is substituted or unsubstituted phenyl; substituted or unsubstituted indolyl; or substituted or unsubstituted indazolyl;

wherein optional substituents at A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , -(CH ₂ ) _i NH ₂ , - (CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₃ is hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _o NHCH ₃ , -(CH ₂ ) _o NH ₂ , -(CH ₂ ) _o N(CH ₃ ) ₂ , - O(CH ₂ ) _o OMe, -O(CH ₂ ) _o OH, -OC(O)CF ₃ , - SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C( O)H or -C(O)NH ₂ ;

i and o are independently integers from 0 to 5;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the present invention relates to a pharmaceutical composition according to the present invention. In another embodiment, the present invention relates to a pharmaceutical composition according to the present invention and at least one pharmaceutically active compound suitable for treating cancer.

In one embodiment, the present invention provides a method of treating, ameliorating, reducing the severity, reducing the risk of, or suppressing cancer, comprising administering a compound of the present invention to a subject with cancer under conditions that are effective in curing the cancer. In another embodiment, the cancer is selected from the group consisting of prostate cancer, drug-resistant prostate cancer, breast cancer, drug-resistant breast cancer, ovarian cancer, drug-resistant ovarian cancer, skin cancer, melanoma, drug-resistant melanoma, lung cancer , colon cancer, glioma, leukemia, lymphoma, kidney cancer, central nervous system cancer, uterine cancer, drug-resistant uterine cancer and combinations thereof. In another embodiment, administration of a compound of the present invention is carried out in combination with another cancer therapy.

In one embodiment, the present invention provides a method of treating a drug-resistant tumor or tumors, comprising administering a compound of the present invention to a subject suffering from cancer under conditions that are effective to cure the drug-resistant tumor or tumors. In another embodiment, the cancer is selected from the group consisting of prostate cancer, drug-resistant prostate cancer, breast cancer, drug-resistant breast cancer, ovarian cancer, drug-resistant ovarian cancer, skin cancer, melanoma, drug-resistant melanoma, lung cancer , colon cancer, glioma, leukemia, lymphoma, kidney cancer, central nervous system cancer, uterine cancer, drug-resistant uterine cancer and combinations thereof. In another embodiment, administration of a compound of the present invention is carried out in combination with another cancer therapy.

Other features and advantages of the present invention will become apparent from the following detailed description, examples and figures. It should be understood, however, that the detailed description and specific examples, while indicating preferred embodiments of the present invention, are given for illustrative purposes only, as various changes and modifications without departing from the spirit and scope of the invention will be apparent. those skilled in the art upon examination of the present detailed description.

Brief description of drawings

Figure 1 depicts the structures of typical tubulin inhibitors that bind to the colchicine binding site.

Figure 2 depicts a synthesis scheme for the preparation of compounds according to the present invention. Reagents and conditions: a) TsOH, EtOH, reflux; b) (3,4,5-trimethoxyphenyl)boronic acid, K ₂ CO ₃ , Pd(PPh ₃ ) ₄ .

Figure 3 shows a synthesis scheme for the preparation of 4-substituted methoxybenzoyl-aryl-thiazole analogues (SMART), aimed at modification at the para position of the benzoyl ring. Reagents and conditions: (a) MeOH/phosphate buffer, pH = 6.4, RT; (b) EDCI, HOBt, NMM, CH ₃ OCH ₃ NH·HCl; (c) CBrCl ₃ , DBU; (d) 5-bromo-1,2,3-trimethoxybenzene/BuLi, THF, -78°C; (e) AlCl ₃ , CH ₂ Cl ₂ ; (f) ClCH ₂ COCl, CH ₂ Cl ₂ , NEt ₃ (12) or (CF ₃ CO) ₂ O, CH ₂ Cl ₂ , DMAP (13); (g) PhCH ₂ Br, K ₂ CO ₃ , DMF (14); MOMCl, Huenig's base, CH ₂ Cl ₂ (15); BrCH ₂ CH ₂ NHBoc, DMF, Cs ₂ CO ₃ (16) or 2-(2-bromoethyl)isoindoline-1,3-dione, K ₂ CO ₃ , DMF, 120°C (17); (h) 4M HCl in dioxane.

Figure 4 shows a synthesis scheme for the preparation of phenyl-amino-thiazole/imidazole analogues. Reagents and conditions: (a) BrCN, Et ₂ O/hexane; (b) CH ₃ SO ₃ H, EtOH, reflux, 24 hours; (c) (1) 6M HCl, (2) NaOH 25% conc.; (d) PhSO ₂ Cl, Et ₃ N; (e) -78°C, t-BuLi, 3,4,5-trimethoxybenzoyl chloride; (f) Boc ₂ O, NaOH; (g) (chloromethanetriyl)tribenzene, Et ₃ N, CH ₂ Cl ₂ ; (h) HCl; (i) H ₂ , Pd-C, 5% EtOH, 40 psi (280 kPa); (j) PhCOSCN, Me ₂ CO; (k) MeOH, 1N. NaOH; (l) EtOH, 65°C; (m) NaOH, MeOH; (n) HBTU, NMM, HNCH ₃ OCH ₃ ⋅HCl, CH ₂ Cl ₂ ; (o) 3,4,5-trimethoxyphenyl bromide, n-BuLi, THF.

Figure 5 depicts tubulin irreversible binding agents and possible Cys-241 interactions at the para position of the benzoyl ring in compounds of formula XI-XV.

Figure 6 shows possible binding sites for compound 4 (tube model, Glidescore docking score -8.58) and 7 (tube model, Glidescore docking score -8.10) in the α,β-tubulin dimer (PDB code: 1SA0) . The natural ligand, DAMA-colchicine (Glidescore docking index -9.26) is shown in a wire model.

Figure 7 depicts possible binding sites for compound 33 (tube model, Glidescore docking score -8.70) and the natural DAMA-colchicine ligand (wire model, Glidescore docking score -9.26) in the α,β-dimer of tubulin (code PDB: 1SA0).

Figure 8 shows the putative metabolites and metabolic pathway of compound 15.

Figure 9 shows the putative metabolites and metabolic pathway of compound 18.

Figure 10 shows the putative metabolites and metabolic pathway of compound 4.

Figure 11 shows the putative metabolites and metabolic pathway of compound 7.

Figure 12 shows the inhibition of tubulin polymerization by compounds 4 and 7 in vitro.

Detailed Description of the Invention

In one embodiment, the present invention relates to a compound of formula (I)

(I)

Where

Q is S, NH or O;

Z represents CH or N;

A represents substituted or unsubstituted systems of single, fused or multiple aryl or (hetero)cyclic rings, including saturated and unsaturated N-heterocycles, saturated and unsaturated S-heterocycles and saturated and unsaturated O-heterocycles, saturated or unsaturated cyclic hydrocarbons, saturated or unsaturated mixed heterocycles, aliphatic linear or branched C ₁ -C ₃₀ hydrocarbons;

wherein optional substituents at A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , - (CH ₂ ) _i NH ₂ , -(CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₁ independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H , NHCO-alkyl, - O(CH ₂ ) _j OCH ₃ , -O(CH ₂ ) _j OH, -O(CH ₂ ) _j NHCH ₃ , -O(CH ₂ ) _j NH ₂ , -O-(CH ₂ ) _j N(CH ₃ ) ₂ , - OC(O)CF ₃ , -OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _j NH ₂ , -O(CH ₂ ) _j N- phthalimide or a combination thereof;

R ₂ represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H, NHCO-alkyl, -O(CH ₂ ) _k OCH ₃ , -O(CH ₂ ) _k OH, -O(CH ₂ ) _k NHCH ₃ , -O(CH ₂ ) _k NH ₂ , -O-(CH ₂ ) _k N(CH ₃ ) ₂ , -OC(O)CF ₃ , - OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _k NH ₂ or -O(CH ₂ ) _k N-phthalimide ;

i, j and k are independently integers from 0 to 5;

n is an integer from 1 to 4;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, tautomer, polymorph, or combination thereof.

In one embodiment, the present invention relates to a compound of formula (II)

(II)

Where

Q is S, NH or O;

Z represents CH or N;

A is substituted or unsubstituted phenyl; substituted or unsubstituted indolyl; or substituted or unsubstituted indazolyl;

wherein optional substituents at A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , - (CH ₂ ) _i NH ₂ , -(CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₁ independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H , NHCO-alkyl, - O(CH ₂ ) _j OCH ₃ , -O(CH ₂ ) _j OH, -O(CH ₂ ) _j NHCH ₃ , -O(CH ₂ ) _j NH ₂ , -O-(CH ₂ ) _j N(CH ₃ ) ₂ , - OC(O)CF ₃ , -OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _j NH ₂ , -O(CH ₂ ) _j N- phthalimide or a combination thereof;

R ₂ represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H, NHCO-alkyl, -O(CH ₂ ) _k OCH ₃ , -O(CH ₂ ) _k OH, -O(CH ₂ ) _k NHCH ₃ , -O(CH ₂ ) _k NH ₂ , -O-(CH ₂ ) _k N(CH ₃ ) ₂ , -OC(O)CF ₃ , - OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _k NH ₂ or -O(CH ₂ ) _k N-phthalimide ;

i, j and k are independently an integer from 0 to 5;

n is an integer from 1 to 4;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the present invention relates to a compound of formula (III)

(III)

Where:

Z represents CH or N;

A is substituted or unsubstituted phenyl; substituted or unsubstituted indolyl; or substituted or unsubstituted indazolyl;

wherein optional substituents at A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , - (CH ₂ ) _i NH ₂ , -(CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₁ independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H , NHCO-alkyl, - O(CH ₂ ) _j OCH ₃ , -O(CH ₂ ) _j OH, -O(CH ₂ ) _j NHCH ₃ , -O(CH ₂ ) _j NH ₂ , -O(CH ₂ ) _j N(CH ₃ ) ₂ , - OC(O)CF ₃ , -OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _j NH ₂ , -O(CH ₂ ) _j N-phthalimide or a combination thereof;

R ₂ represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H, NHCO-alkyl, -O(CH ₂ ) _k OCH ₃ , -O(CH ₂ ) _k OH, -O(CH ₂ ) _k NHCH ₃ , -O(CH ₂ ) _k NH ₂ , -O(CH ₂ ) _k N(CH ₃ ) ₂ , -OC(O)CF ₃ , - OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _k NH ₂ or -O(CH ₂ ) _k N-phthalimide;

i, j and k are independently integers from 0 to 5;

n is an integer from 1 to 4;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the present invention relates to a compound of formula (IV)

(IV)

Where

A is substituted or unsubstituted phenyl; substituted or unsubstituted indolyl; or substituted or unsubstituted indazolyl;

wherein optional substituents at A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , - (CH ₂ ) _i NH ₂ , -(CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₁ independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H , NHCO-alkyl, - O(CH ₂ ) _j OCH ₃ , -O(CH ₂ ) _j OH, -O(CH ₂ ) _j NHCH ₃ , -O(CH ₂ ) _j NH ₂ , -O(CH ₂ ) _j N(CH ₃ ) ₂ , - OC(O)CF ₃ , -OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _j NH ₂ , -O(CH ₂ ) _j N-phthalimide or a combination thereof;

R ₂ represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H, NHCO-alkyl, -O(CH ₂ ) _k OCH ₃ , -O(CH ₂ ) _k OH, -O(CH ₂ ) _k NHCH ₃ , -O(CH ₂ ) _k NH ₂ , -O(CH ₂ ) _k N(CH ₃ ) ₂ , -OC(O)CF ₃ , - OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _k NH ₂ or -O(CH ₂ ) _k N-phthalimide;

i, j and k are independently integers from 0 to 5;

n is an integer from 1 to 4;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the present invention relates to a compound of formula (V)

(V)

Where

A is substituted or unsubstituted phenyl; substituted or unsubstituted indolyl; or substituted or unsubstituted indazolyl;

wherein optional substituents at A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , - (CH ₂ ) _i NH ₂ , -(CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₁ independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H , NHCO-alkyl, - O(CH ₂ ) _j OCH ₃ , -O(CH ₂ ) _j OH, -O(CH ₂ ) _j NHCH ₃ , -O(CH ₂ ) _j NH ₂ , -O(CH ₂ ) _j N(CH ₃ ) ₂ , - OC(O)CF ₃ , -OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _j NH ₂ , -O(CH ₂ ) _j N-phthalimide or a combination thereof;

R ₂ represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H, NHCO-alkyl, -O(CH ₂ ) _k OCH ₃ , -O(CH ₂ ) _k OH, -O(CH ₂ ) _k NHCH ₃ , -O(CH ₂ ) _k NH ₂ , -O(CH ₂ ) _k N(CH ₃ ) ₂ , -OC(O)CF ₃ , - OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _k NH ₂ or -O(CH ₂ ) _k N-phthalimide;

i, j and k are independently integers from 0-5;

n is an integer from 1 to 4;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the present invention relates to a compound of formula (VI)

(VI)

Where

Q is NH, S or O;

Z represents CH or N;

A is substituted or unsubstituted phenyl; substituted or unsubstituted indolyl; or substituted or unsubstituted indazolyl;

wherein optional substituents at A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , - (CH ₂ ) _i NH ₂ , -(CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₂ represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H, NHCO-alkyl, -O(CH ₂ ) _k OCH ₃ , -O(CH ₂ ) _k OH, -O(CH ₂ ) _k NHCH ₃ , -O(CH ₂ ) _k NH ₂ , -O(CH ₂ ) _k N(CH ₃ ) ₂ , -OC(O)CF ₃ , - OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _k NH ₂ or -O(CH ₂ ) _k N-phthalimide;

i and k are independently integers from 0 to 5;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the present invention relates to a compound of formula (VII)

(VII)

Where

Z represents CH or N;

A is substituted or unsubstituted phenyl; substituted or unsubstituted indolyl; or substituted or unsubstituted indazolyl;

wherein optional substituents at A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , - (CH ₂ ) _i NH ₂ , -(CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

i is an integer from 0 to 5;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the present invention relates to a compound of formula (VIII)

(VIII)

Where

Z represents CH or N;

R ₄ independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _q NHCH ₃ , -(CH ₂ ) _q NH ₂ , - (CH ₂ ) _q N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₅ is hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _t NHCH ₃ , -(CH ₂ ) _t NH ₂ , -(CH ₂ ) _t N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH ₂ ;

m is an integer from 1 to 4;

t and q are independently integers from 0 to 5;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the compound of Formula VIII is:

3, where its tautomer is:

In one embodiment, the present invention relates to a compound of formula (IX)

(IX)

Where

Z represents CH or N;

R ₄ independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _q NHCH ₃ , -(CH ₂ ) _q NH ₂ , - (CH ₂ ) _q N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₅ is hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _t NHCH ₃ , -(CH ₂ ) _t NH ₂ , -(CH ₂ ) _t N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH ₂ ;

m is an integer from 1 to 4;

t and q are independently integers from 0 to 5;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the compound of formula IX is:

, where its tautomer is: ; or

, where its tautomer is: .

In one embodiment, the present invention provides a compound of formula (X)

(X)

Where

Z represents CH or N;

P represents CH or N;

R ₄ independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _q NHCH ₃ , -(CH ₂ ) _q NH ₂ , - (CH ₂ ) _q N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₅ is hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _t NHCH ₃ , -(CH ₂ ) _t NH ₂ , -(CH ₂ ) _t N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH ₂ ;

m is an integer from 1 to 4;

t and q are independently integers from 0 to 5;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the compound of formula X is:

5, where its tautomer is: , or

6, where its tautomer includes: .

In one embodiment, the present invention relates to a compound of formula (XI)

(XI)

Where

Q is S, NH or O;

X represents a bond or NH;

A represents substituted or unsubstituted systems of single, fused or multiple aryl or (hetero)cyclic rings, including saturated and unsaturated N-heterocycles, saturated and unsaturated S-heterocycles and saturated and unsaturated O-heterocycles, saturated or unsaturated cyclic hydrocarbons, saturated or unsaturated mixed heterocycles, aliphatic linear or branched C ₁ -C ₃₀ hydrocarbons;

wherein optional substituents at A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , -(CH ₂ ) _i NH ₂ , - (CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R represents hydrogen, C ₁ -C ₅ linear or branched alkyl; C ₁ -C ₅ linear or branched alkenyl, (CH ₂ ) _l Ph, aryl, (CH ₂ ) _l NHCH ₃ , -(CH ₂ ) _l NH ₂ , -(CH ₂ ) _l N(CH ₃ ) _2, C (O)alkyl, C(O)CF ₃ , -(CH ₂ ) _l OMe, -(CH ₂ ) _l OH or haloalkyl;

R₃ represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO₂, haloalkyl, CF₃,CN,-CH₂CN, NH₂, hydroxyl, -O(CH₂)_oNHCH₃, -O(CH₂)_oN.H.₂, -O(CH₂)_oN(CH₃)₂, - O(CH₂)_oOMe, -O(CH₂)_oOH, -OC(O)CF₃, -SO₂-aryl, C₁-C₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH₂Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH₂;

i, l and o are independently integers from 0 to 5;

or a metabolite, isomer, N-oxide, prodrug, hydrate, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the present invention relates to a compound of formula (XII)

(XII)

Where

Q is S, NH or O;

X represents a bond or NH;

A is substituted or unsubstituted phenyl; substituted or unsubstituted indolyl; or substituted or unsubstituted indazolyl;

wherein optional substituents at A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , -(CH ₂ ) _i NH ₂ , - (CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R₃ represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO₂, haloalkyl, CF₃,CN,-CH₂CN, NH₂, hydroxyl, -O(CH₂)_oNHCH₃, -O(CH₂)_oN.H.₂, -O(CH₂)_oN(CH₃)₂, - O(CH₂)_oOMe, -O(CH₂)_oOH, -OC(O)CF₃, -SO₂-aryl, C₁-C₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH₂Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH₂;

i and o are independently integers from 0 to 5;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment of Formula XII, R ₃ is different from OMe.

In one embodiment, the present invention relates to a compound of formula (XIII)

(XIII)

Where

Q is S, NH or O;

R represents hydrogen, C₁-C₅ linear or branched alkyl; C₁-C₅ linear or branched alkenyl, (CH₂)_lPh, aryl, (CH₂)_lNHCH₃, -(CH₂)_lN.H.₂, -(CH₂)_lN(CH₃)₂, C(O)alkyl, C(O)CF₃, -(CH₂)_lOme, -(CH₂)_lOH or haloalkyl;

R₃ represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO₂, haloalkyl, CF₃,CN,-CH₂CN, NH₂, hydroxyl, -O(CH₂)_oNHCH₃, -O(CH₂)_oN.H.₂, -O(CH₂)_oN(CH₃)₂, - O(CH₂)_oOMe, -O(CH₂)_oOH, -OC(O)CF₃, -SO₂-aryl, C₁-C₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH₂Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH₂;

R ₄ independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _q NHCH ₃ , -(CH ₂ ) _q NH ₂ , - (CH ₂ ) _q N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₅ is hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _t NHCH ₃ , -(CH ₂ ) _t NH ₂ , -(CH ₂ ) _t N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH ₂ ;

m is an integer from 1 to 4;

l, o, t and q are independently integers from 0 to 5;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment, the compound of formula XIII is:

; 14; 15; or 33.

In one embodiment, the present invention provides a compound of formula (XIV):

(XIV)

Where

Q is S, NH or O;

R represents hydrogen, C₁-C₅ linear or branched alkyl, C₁-C₅ linear or branched alkenyl, (CH₂)_lPh, aryl, (CH₂)_lNHCH₃, -(CH₂)_lN.H.₂, -(CH₂)_lN(CH₃)₂, C(O)alkyl, C(O)CF₃, -(CH₂)_lOMe, -(CH₂)_lOH or haloalkyl;

R₃ represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO₂, haloalkyl, CF₃,CN,-CH₂CN, NH₂, hydroxyl, -O(CH₂)_oNHCH₃, -O(CH₂)_oN.H.₂, -O(CH₂)_oN(CH₃)₂, - O(CH₂)_oOMe, -O(CH₂)_oOH, -OC(O)CF₃, -SO₂-aryl, C₁-C₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH₂Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH₂;

R ₄ independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _q NHCH ₃ , -(CH ₂ ) _q NH ₂ , - (CH ₂ ) _q N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₅ is hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _t NHCH ₃ , -(CH ₂ ) _t NH ₂ , -(CH ₂ ) _t N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH ₂ ;

m is an integer from 1 to 4;

l, o, t and q are independently integers from 0 to 5;

or a metabolite, hydrate, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer or combination thereof.

In one embodiment, the present invention relates to a compound of formula (XV)

(XV)

Where

Q is S, NH or O;

X represents a bond or NH;

R represents hydrogen, C₁-C₅ linear or branched alkyl, C₁-C₅ linear or branched alkenyl, (CH₂)_lPh, aryl, (CH₂)_lNHCH₃, -(CH₂)_lN.H.₂, -(CH₂)_lN(CH₃)₂, C(O)alkyl, C(O)CF₃, -(CH₂)_lOme, -(CH₂)_lOH or haloalkyl;

R₃ represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO₂, haloalkyl, CF₃,CN,-CH₂CN, NH₂, hydroxyl, -O(CH₂)_oNHCH₃, -O(CH₂)_oN.H.₂, -O(CH₂)_oN(CH₃)₂, - O(CH₂)_oOMe, -O(CH₂)_oOH, -OC(O)CF₃, -SO₂-aryl, C₁-C₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH₂Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH₂;

R ₄ independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _q NHCH ₃ , -(CH ₂ ) _q NH ₂ , - (CH ₂ ) _q N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof;

R ₅ is hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _t NHCH ₃ , -(CH ₂ ) _t NH ₂ , -(CH ₂ ) _t N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino , aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH ₂ ;

m is an integer from 1 to 4;

l, o, t and q are independently integers from 0 to 5;

or a metabolite, hydrate, isomer, N-oxide, prodrug, pharmaceutical product, pharmaceutically acceptable salt, polymorph, tautomer, or combination thereof.

In one embodiment of formula XI, XIII, XIV and XV, OR is different from R ₃ .

In one embodiment, compounds of formula XI-XV contain an R ₃ group at the para position of the benzoyl ring. It was found that the introduction of different functional groups at the para position of the benzoyl ring likely provides stronger interactions with Cys-241 in the β-tubulin subunit (Figure 5), thereby increasing the tubulin polymerization inhibition activity. In addition, modification of the benzoyl ring helped to understand the possible mechanism of metabolic demethylation. Hydrophobic (OBn 14, OMOM 15) and hydrophilic (OCH ₂ CH ₂ NH ₂ , 18) groups were introduced, as shown in Figure 3.

Another strategy was proposed based on the hypothesis that when an alkylating group is introduced at the para position of the benzoyl ring, it can form an irreversible covalent bond with the mercapto group of Cys-241 in the colchicine binding domain and provide irreversible mitotic blocks. The mechanism of inhibition of microtubule assembly upon binding of small molecule drugs to tubulin through covalent interaction with the amino residue of tubulin has been well described. Bai et al. [Bai, R.; Covell, D. G.; Pei, X. F.; Ewell, J. B.; Nguyen, N. Y.; Brossi, A.; Hamel, E., Mapping the binding site of colchicinoids on beta-tubulin. 2-Chloroacetyl-2-demethylthiocolchicine covalently reacts predominantly with cysteine 239 and secondarily with cysteine 354. J Biol Chem 2000, 275(51), 40443-52] reported that 2- and 3-chloroacetyl analogs of dimethylthiocolchicine bind irreversibly to the colchicine binding site , mainly at Cys-241 and prevent colchicine-binding agents from binding to this site. Covalent interaction of 2,4-dichlorobenzylthiocyanate (Figure 5) with tubulin occurs at several cysteine residues, in particular Cys-241 of β-tubulin [Bai, R. L.; Lin, C. M.; Nguyen, N. Y.; Liu, T. Y.; Hamel, E., Identification of the cysteine residue of beta-tubulin alkylated by the antimitotic agent 2,4-dichlorobenzyl thiocyanate, facilitated by separation of the protein subunits of tubulin by hydrophobic column chromatography. Biochemistry 1989, 28(13), 5606-12]. It is believed that the formation of a covalent bond between tubulin and the 2,4-dichlorobenzylmercaptan fragment is reversible. 2-fluoro-1-methoxy-4-(pentafluorophenylsulfonamido)benzene (T138067, Figure 5) irreversibly binds β-tubulin by replacing the F atom in the para position with a Cys-241 thiol group. This compound ensured the recruitment of unmodified tubulin dimers into large amorphous aggregates and, thus, quickly consumed the supply of tubulin available for microtubule formation [Shan, B.; Medina, J.C.; Santha, E.; Frankmoelle, W. P.; Chou, T. C.; Learned, R. M.; Narbut, M. R.; Stott, D.; Wu, P.; Jaen, J. C.; Rosen, T.; Timmermans, P. B.; Beckmann, H., Selective, covalent modification of beta-tubulin residue Cys-239 by T138067, an antitumor agent with in vivo efficacy against multidrug-resistant tumors. Proceedings of the National Academy of Sciences of the United States of America 1999, 96(10), 5686-91].

In one embodiment, the compounds of the present invention incorporate an alkylating functional group to form a covalent bond or enhance the interaction between Cys-241 and the benzoyl ring of the compounds of the present invention. Thus, the chloroacetic analogue (12) and trifluoroacetate (13), shown in Figure 3, were also synthesized and studied.

In one embodiment, A in Formula I and XI is a substituted or unsubstituted system of single, fused or multiple aryl or (hetero)cyclic rings, including saturated and unsaturated N-heterocycles, saturated and unsaturated S-heterocycles, and saturated and unsaturated O- heterocycles, saturated or unsaturated cyclic hydrocarbons, saturated or unsaturated mixed heterocycles, aliphatic linear or branched C ₁ -C ₃₀ hydrocarbons.

In one embodiment, group A is substituted or unsubstituted furanyl, benzofuranyl, benzothiophenyl, indolyl, indazolyl, pyridinyl, phenyl, biphenyl, triphenyl, diphenylmethane, adamantanyl, flurenyl, or other heterocyclic analogs such as, for example, pyrrolyl, pyrazolyl, imidazolyl . ranil, tetrahydropyranyl, dioxanyl, furanyl, pyrilium .

As used herein, "saturated or unsaturated cyclic hydrocarbons" may be any specified cyclic hydrocarbon, including, but not limited to, phenyl, biphenyl, triphenyl, naphthyl, cycloalkyl, cycloalkenyl, cyclodienyl, fluorene, adamantane, etc.; "saturated or unsaturated N-heterocycles" may be any specified N-containing heterocycle, including, but not limited to, aza- and diazacycloalkyls such as aziridinyl, azetidinyl, diazatidinyl, pyrrolidinyl, piperidinyl, piperazinyl and azocanyl, pyrrolyl, pyrazolyl, imidazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl, pyrrolizinyl, indolyl, indazolyl, quinolinyl, isoquinolinyl, benzimidazolyl, indazolyl, quinolizinyl, cinnolinyl, quinalolinyl, phthalazinyl, naphthyridinyl, quinoxalinyl, etc.; "saturated or unsaturated O-heterocycles" may be any specified O-containing heterocycle, including, but not limited to, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, dioxanyl, furanyl, pyrilium, benzofuranyl, benzodioxolyl, etc.; "saturated or unsaturated S-heterocycles" can be any specified S-containing heterocycle, including, but not limited to, tyranyl, thietanyl, tetrahydrothiophenyl, dithiolanyl, tetrahydrothiopyranyl, thiophenyl, thiepinyl, tianaphenyl, etc.; "saturated or unsaturated mixed heterocycles" can be any heterocycle containing two or more S, N or O heteroatoms, including, but not limited to, oxathiolanyl, morpholinyl, thioxanyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, etc. .d.

In one embodiment, A in formula II-VII and XII is substituted or unsubstituted phenyl; substituted or unsubstituted indolyl; or substituted or unsubstituted indazolyl. In another embodiment, A in formula I, II-VII, XI and XII is substituted or unsubstituted phenyl. In another embodiment, A in formula I, II-VII, XI and XII is substituted or unsubstituted indolyl. In another embodiment, the indolyl is a substituted or unsubstituted 3-indolyl, 4-indolyl, or 5-indolyl. In another embodiment, group A is a substituted or unsubstituted 3-indolyl. In certain embodiments, group A is unsubstituted 3-indolyl. In certain embodiments, group A is unsubstituted 5-indolyl. In certain embodiments, group A is a substituted 5-indolyl. In another embodiment, indolyl is substituted or unsubstituted and is selected from the following structures:

, , or .

In another embodiment, A in formula I, II-VII, XI and XII is substituted or unsubstituted indazolyl. In another embodiment, indazolyl is a substituted or unsubstituted 3-indazolyl, 4-indazolyl, or 5-indazolyl. In another embodiment, group A is a substituted or unsubstituted 3-indazolyl. In certain embodiments, group A is unsubstituted 3-indazolyl. In certain embodiments, group A is unsubstituted 5-indazolyl. In certain embodiments, group A is a substituted 5-indazolyl. In another embodiment, indazolyl is substituted or unsubstituted and is selected from the following structures:

, or .

In one embodiment, the optional substituents on A in formula I, II-VII, XI and XII are absent or include one, two, three, four or five substituents. In another embodiment, the substituents are the same. In another embodiment, the substituents are different. Individual substituents may be present in the ortho, meta or para positions. In some embodiments, when there are two or more substituents, one of them is in the para position.

In other embodiments, optional substituents on A include O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, -(CH ₂ ) _i NHCH ₃ , -(CH ₂ ) _i NH ₂ , -(CH ₂ ) _i N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ , NO ₂ or their combination. In another embodiment, A is unsubstituted. In another embodiment, A is substituted with O-alkyl. In another embodiment, A is substituted with O-haloalkyl. In another embodiment, A is substituted with halogen. In another embodiment, A is replaced with NO ₂ . In another embodiment, A is substituted with haloalkyl. In another embodiment, A is replaced by CF ₃ . In another embodiment, A is replaced by CN. In another embodiment, A is substituted with -CH ₂ CN. In another embodiment, A is substituted with NH ₂ . In another embodiment, A is replaced with hydroxyl. In another embodiment, A is substituted - (CH ₂ ) _i NHCH ₃ . In another embodiment, A is substituted with -(CH ₂ ) _i NH ₂ . In another embodiment, A is substituted with -(CH ₂ ) _i N(CH ₃ ) ₂ . In another embodiment, A is substituted with OC(O)CF ₃ . In another embodiment, A is substituted with -SO ₂ -aryl. In another embodiment, A is substituted with C ₁ -C ₅ linear or branched alkyl. In another embodiment, A is substituted with haloalkyl. In another embodiment, A is substituted with alkylamino. In another embodiment, A is substituted with aminoalkyl. In another embodiment, A is substituted with -OCH ₂ Ph. In another embodiment, A is substituted with -NHCO-alkyl. In another embodiment, A is replaced with COOH. In another embodiment, A is replaced with -C(O)Ph. In another embodiment, A is substituted with C(O)O-alkyl. In another embodiment, A is substituted with C(O)H. In another embodiment, A is substituted with - C(O)NH ₂ .

In one embodiment, i is an integer between 0 and 5. In another embodiment, i is 0. In another embodiment, i is 1. In another embodiment, i is 2. In another embodiment, i is 3. In another embodiment implementation i is 4. In another implementation i is 5.

In one embodiment, Z in Formula I-III and VI-X is CH or N. In another embodiment, Z is CH. In another embodiment, Z is N.

In one embodiment, P in formula X is CH or N. In another embodiment, P is CH. In another embodiment, P is N.

In one embodiment, Q in formula I, II, VI, XI, XII and XIII is S, O or NH. In another embodiment, Q is S. In another embodiment, Q is O. In another embodiment, Q is NH.

In one embodiment, X in formula XI and XV is a bond or NH. In another embodiment, X represents a relationship. In another embodiment, X is NH.

In one embodiment, R ₁ in Formula IV is hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ , hydroxyl, COOH, C(O)H, NHCO-alkyl, -O(CH ₂ ) _j OCH ₃ , -O(CH ₂ ) _j OH, -O(CH ₂ ) _j NHCH ₃ , -O(CH ₂ ) _j NH ₂ , - O-(CH ₂ ) _j N(CH ₃ ) ₂ , -OC(O)CF ₃ , -OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _j NH ₂ or -O (CH ₂ ) _j N-phthalimide. In another embodiment, R ₁ is hydrogen. In another embodiment, R ₁ represents O-alkyl. In another embodiment, R ₁ is O-haloalkyl. In another embodiment, R ₁ is halogen. In another embodiment, R ₁ is an alkyl halo. In another embodiment, R ₁ is CF ₃ . In another embodiment, R ₁ is CN. In another embodiment, R ₁ is NO ₂ . In another embodiment, R ₁ is -CH ₂ CN. In another embodiment, R ₁ is NH ₂ . In another embodiment, R ₁ represents hydroxyl. In another embodiment, R ₁ represents COOH. In another embodiment, R ₁ is C(O)H. In another embodiment, R ₁ is NHCO-alkyl. In another embodiment, R ₁ is -O(CH ₂ ) _j OCH ₃ . In another embodiment, R ₁ is -O(CH ₂ ) _j OH. In another embodiment, R ₁ is -O(CH ₂ ) _j NHCH ₃ . In another embodiment, R ₁ is -O(CH ₂ ) _j NH ₂ . In another embodiment, R ₁ is -O-(CH ₂ ) _j N(CH ₃ ) ₂ . In another embodiment, R ₁ is -OC(O)CF ₃ . In another embodiment, R ₁ is -OC(O)CH ₂ Cl. In another embodiment, R ₁ is -OCH ₂ Ph. In another embodiment, R ₁ is - O(CH ₂ ) _j NH ₂ . In another embodiment, R ₁ is -O(CH ₂ ) _j N-phthalimide.

In one embodiment, j is an integer between 0 and 5. In another embodiment, j is 0. In another embodiment, j is 1. In another embodiment, j is 2. In another embodiment, j is 3. In another embodiment implementation j is equal to 4. In another implementation, j is equal to 5.

In one embodiment, n is an integer from 1 to 4. In another embodiment, n is 1. In another embodiment, n is 2; in another embodiment, n is 3. In another embodiment, n is 4.

In one embodiment, R ₂ in formula I-VI is hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, haloalkyl, CF ₃ , CN, NO ₂ , -CH ₂ CN, NH ₂ . hydroxyl, COOH, C(O)H, NHCO-alkyl, -O(CH ₂ ) _j OCH ₃ , -O(CH ₂ ) _j OH, -O(CH ₂ ) _j NHCH ₃ , - O(CH ₂ ) _j NH ₂ , -O-(CH ₂ ) _j N(CH ₃ ) ₂ , -OC(O)CF ₃ , -OC(O)CH ₂ Cl, -OCH ₂ Ph, -O(CH ₂ ) _j NH ₂ or - O(CH ₂ ) _j N-phthalimide. In another embodiment, R ₂ is hydrogen. In another embodiment, R ₂ represents O-alkyl. In another embodiment, R ₂ is O-haloalkyl. In another embodiment, R ₂ is halogen. In another embodiment, R ₂ is an alkyl halo. In another embodiment, R ₂ is CF ₃ . In another embodiment, R ₂ is CN. In another embodiment, R ₂ is NO ₂ . In another embodiment, R ₂ is -CH ₂ CN. In another embodiment, R ₂ is NH ₂ . In another embodiment, R ₂ represents hydroxyl. In another embodiment, R ₁ represents COOH. In another embodiment, R ₂ is C(O)H. In another embodiment, R ₂ is NHCO-alkyl. In another embodiment, R ₂ is -O(CH ₂ ) _k OCH ₃ . In another embodiment, R ₂ is -O(CH ₂ ) _k OH. In another embodiment, R ₂ is -O(CH ₂ ) _k NHCH ₃ . In another embodiment, R ₂ is -O(CH ₂ ) _k NH ₂ . In another embodiment, R ₂ is - O-(CH ₂ ) _k N(CH ₃ ) ₂ . In another embodiment, R ₂ is -OC(O)CF ₃ . In another embodiment, R ₂ is -OC(O)CH ₂ Cl. In another embodiment, R ₂ is -OCH ₂ Ph. In another embodiment, R ₂ is - O(CH ₂ ) _k NH ₂ . In another embodiment, R ₂ is -O(CH ₂ ) _k N-phthalimide.

In one embodiment, at least one of R ₁ and R ₂ in Formula IV is not hydrogen.

In one embodiment, k is an integer from 0 to 5. In another embodiment, k is 0. In another embodiment, k is 1. In another embodiment, k is 2. In another embodiment, k is 3. In another embodiment implementation k is 4. In another implementation k is 5.

In one implementation of R₃ in formula XI-XV represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO₂, haloalkyl, CF₃,CN,-CH₂CN, NH₂, hydroxyl, -O(CH₂)_oNHCH₃, -O(CH₂)_oN.H.₂, -O(CH₂)_oN(CH₃)₂, -O(CH₂)_oOMe, -O(CH₂)_oOH, - OC(O)CF₃, -SO₂-aryl, C₁-C₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH₂Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O)NH₂. In another implementation of R₃ represents hydrogen. In another implementation of R₃ represents O-alkyl. In another implementation of R₃ is O-haloalkyl. In another implementation of R₃ is a halogen. In another implementation of R₃ represents NO₂. In another implementation of R₃ is a haloalkyl. In another implementation of R₃ represents CF₃. In another implementation of R₃ represents CN. In another implementation of R₃ represents -CH₂CN. In another implementation of R₃ represents NH₂. In another implementation of R₃ represents hydroxyl. In another implementation of R₃ represents -O(CH₂)_oNHCH₃. In another implementation of R₃ represents -O(CH₂)_oN.H.₂. In another implementation of R₃ represents -O(CH₂)_oN(CH₃)₂. In another implementation of R₃ represents -O(CH₂)_oOme. In another implementation of R₃ represents - O(CH₂)_oOH. In another implementation of R₃ represents -OC(O)CF₃. In another implementation of R₃ represents -SO₂-aryl. In another implementation of R₃ represents C₁-C₅ linear or branched alkyl. In another implementation of R₃ is a haloalkyl. In another implementation of R₃ is an alkylamino. In another implementation of R₃ is an aminoalkyl. In another implementation of R₃ represents -OCH₂Ph. In another implementation of R₃ represents -NHCO-alkyl. In another implementation of R₃ represents COOH. In another implementation of R₃ represents - C(O)Ph. In another implementation of R₃ is C(O)O-alkyl. In another implementation of R₃ represents C(O)H. In another implementation of R₃ represents -C(O)NH₂.

In one embodiment, o is an integer between 0 and 5. In another embodiment, o is 0. In another embodiment, o is 1. In another embodiment, o is 2. In another embodiment, o is 3. In another embodiment implementation o is 4. In another embodiment, o is 5. In one embodiment, R ₄ in formula VIII-X and XIII-XV independently represents hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN, NH ₂ , hydroxyl, - (CH ₂ ) _q NHCH ₃ , -(CH ₂ ) _q NH ₂ , -(CH ₂ ) _q N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl, C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H, -C(O)NH ₂ or a combination thereof. In another embodiment, R ₄ is hydrogen. In another embodiment, R ₄ is O-alkyl. In another embodiment, R ₄ is O-haloalkyl. In another embodiment, R ₄ is halogen. In another embodiment, R ₄ is NO ₂ . In another embodiment, R ₄ is an alkyl halo. In another embodiment, R ₄ is CF ₃ . In another embodiment, R ₄ is CN. In another embodiment, R ₄ is -CH ₂ CN. In another embodiment, R ₄ is NH ₂ . In another embodiment, R ₄ is hydroxyl. In another embodiment, R ₄ is -(CH ₂ ) _q NHCH ₃ . In another embodiment, R ₄ is - (CH ₂ ) _q NH ₂ . In another embodiment, R ₄ is -(CH ₂ ) _q N(CH ₃ ) ₂ . In another embodiment, R ₄ is -OC(O)CF ₃ . In another embodiment, R ₄ is -SO ₂ -aryl. In another embodiment, R ₄ is a C ₁ -C ₅ linear or branched alkyl. In another embodiment, R ₄ is an alkyl halo. In another embodiment, R ₄ is alkylamino. In another embodiment, R ₄ is aminoalkyl. In another embodiment, R ₄ is -OCH ₂ Ph. In another embodiment, R ₄ is -NHCO-alkyl. In another embodiment, R ₄ is COOH. In another embodiment, R ₄ is - C(O)Ph. In another embodiment, R ₄ is C(O)O-alkyl. In another embodiment, R ₄ is C(O)H. In another embodiment, R ₄ is -C(O)NH ₂ .

In one embodiment, q is an integer between 0 and 5. In another embodiment, q is 0. In another embodiment, q is 1. In another embodiment, q is 2. In another embodiment, q is 3. in another embodiment implementation, q is 4. In another implementation, q is 5. In one implementation, m is an integer between 1 and 4. In another implementation, m is 1. In another implementation, m is 2. In another implementation, m is 3. In another implementation, m is equal to 4.

In one embodiment, R ₅ in formula VIII-X and XIII-XV is hydrogen, O-alkyl, O-haloalkyl, F, Cl, Br, I, NO ₂ , haloalkyl, CF ₃ , CN, -CH ₂ CN , NH ₂ , hydroxyl, -(CH ₂ ) _q NHCH ₃ , -(CH ₂ ) _q NH ₂ , -(CH ₂ ) _q N(CH ₃ ) ₂ , -OC(O)CF ₃ , -SO ₂ -aryl , C ₁ -C ₅ linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH ₂ Ph, -NHCO-alkyl, COOH, -C(O)Ph, C(O)O-alkyl, C(O)H or -C(O) _NH2 . In another embodiment, R ₅ is hydrogen. In another embodiment, R ₅ represents O-alkyl. In another embodiment, R ₅ is O-haloalkyl. In another embodiment, R ₅ is halogen. In another embodiment, R ₅ is NO ₂ . In another embodiment, R ₅ is an alkyl halo. In another embodiment, R ₅ is CF ₃ . In another embodiment, R ₅ is CN. In another embodiment, R ₅ is -CH ₂ CN. In another embodiment, R ₅ is NH ₂ . In another embodiment, R ₅ is hydroxyl. In another embodiment, R ₅ is -(CH ₂ ) _t NHCH ₃ . In another embodiment, R ₅ is - (CH ₂ ) _t NH ₂ . In another embodiment, R ₅ is -(CH ₂ ) _t N(CH ₃ ) ₂ . In another embodiment, R ₅ is -OC(O)CF ₃ . In another embodiment, R ₅ is -SO ₂ -aryl. In another embodiment, R ₅ is a C ₁ -C ₅ linear or branched alkyl. In another embodiment, R ₅ is an alkyl halo. In another embodiment, R ₅ is alkylamino. In another embodiment, R ₅ is aminoalkyl. In another embodiment, R ₅ is -OCH ₂ Ph. In another embodiment, R ₅ is NHCO-alkyl. In another embodiment, R ₅ is COOH. In another embodiment, R ₅ is -C(O)Ph. In another embodiment, R ₅ is C(O)O-alkyl. In another embodiment, R ₅ is C(O)H. In another embodiment, R ₅ is -C(O)NH ₂ .

In one embodiment, t is an integer from 0 to 5. In another embodiment, t is 0. In another embodiment, t is 1. In another embodiment, t is 2. In another embodiment, t is 3. In another embodiment implementation t is 4. In another implementation t is 5.

In one embodiment, R in formula XI, XIII-XV is hydrogen, C₁-C₅ linear or branched alkyl; C₁-C₅ linear or branched alkenyl, (CH₂)_lPh, aryl, (CH₂)_lNHCH₃, -(CH₂)_lN.H.₂, -(CH₂)_lN(CH₃)₂, C(O)alkyl, C(O)CF₃, - (CH₂)_lOme, -(CH₂)_lOH or haloalkyl. In another embodiment, R is hydrogen. In another embodiment, R is C₁-C₅ linear or branched alkyl. In another embodiment, R is C₁-C₅ linear or branched alkenyl. In another embodiment, R is (CH₂)_lPh. In another embodiment, R is aryl. In another embodiment, R is (CH₂)_lNHCH₃. In another embodiment, R is -(CH₂)_lN.H.₂. In another embodiment, R is - (CH₂)_lN(CH₃)₂. In another embodiment, R is C(O)alkyl. In another embodiment, R is C(O)CF₃. In another embodiment, R is -(CH₂)_lOme. In another embodiment, R is - (CH₂)_lOH. In another embodiment, R is an alkyl halo.

In one embodiment, l is an integer from 0 to 5. In another embodiment, l is 0. In another embodiment, l is 1. In another embodiment, l is 2. In another embodiment, l is 3. In another embodiment implementation l is 4. In another implementation, l is 5.

As used herein, the term “alkyl” can be any straight or branched alkyl group containing up to 30 carbon atoms unless specifically stated otherwise. In another embodiment, alkyl is C ₁ -C ₅ alkyl. In another embodiment, alkyl is C ₂ -C ₅ alkyl. In another embodiment, alkyl is C ₁ -C ₇ alkyl. The alkyl group may be a separate substituent or may be a component of a larger substituent such as alkoxy, haloalkyl, arylalkyl, alkylamino, dialkylamino, alkylamido, alkylurea, etc. Preferred alkyl groups are methyl, ethyl and propyl and thus halomethyl, dihalomethyl, trihalomethyl, haloethyl, dihaloethyl, trihaloethyl, halopropyl, dihalopropyl, trihalopropyl, methoxy, ethoxy, propoxy, arylmethyl, arylethyl, arylpropyl, methylamino, ethylamino, propylamino, dimethylamino, diethylamino, methylamido, acetamido, propylamido, halomethylamido, haloethylamido, halopropylamido, methylurea, ethylurea, propylurea, etc.

As used herein, the term “aryl” refers to any aromatic ring substituent that is directly bonded to the ring member(s) of R ¹ or R ² . The aryl group may be a separate substituent, or the aryl group may be a component of a larger substituent such as arylalkyl, arylamino, arylamido, etc. Exemplary aryl groups include, but are not limited to, phenyl, toluyl, xylyl, furanyl, naphthyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, thiazolyl, oxazolyl, isoxazolyl, pyrazolyl, imidazolyl, thiophenyl, pyrrolyl, phenylmethyl, phenylethyl, phenylamino, phenylamido, etc. d.

As used herein, the term "aminoalkyl" refers to an amino group substituted with an alkyl group, such as defined above. Aminoalkyl refers to a monoalkylamine, dialkylamine or trialkylamine. Non-limiting examples of aminoalkyl groups include -N(Me) ₂ , -NHMe, -NH ₃ .

A "haloalkyl" group refers, in another embodiment, to an alkyl group, as defined above, substituted with one or more halogen atoms, for example, F, Cl, Br or I. Non-limiting examples of haloalkyl groups include CF ₃ , CF ₂ CF ₃ , CH ₂ CF ₃ .

In one embodiment, the invention provides a compound of the present invention or a metabolite thereof, a pharmaceutically acceptable salt, pharmaceutical product, hydrate, isomer, N-oxide, prodrug, tautomer, polymorph, or combinations thereof. In one embodiment, the present invention provides a metabolite of a compound of the present invention. In another embodiment, the present invention provides a pharmaceutically acceptable salt of a compound of the present invention. In another embodiment, the present invention provides a pharmaceutical product of a compound of the present invention. In another embodiment, the present invention provides a hydrate of the compound of the present invention. In another embodiment, the present invention provides an isomer of the compound of the present invention. In another embodiment, the present invention provides an N-oxide of a compound of the present invention. In another embodiment, the present invention provides a prodrug of a compound of the present invention. In another embodiment, the present invention provides a tautomer of a compound of the present invention. In another embodiment, the present invention provides a polymorph of a compound of the present invention. In another embodiment, the present invention provides a composition containing a compound of the present invention, such as described herein, or in another embodiment, a combination of a metabolite, pharmaceutically acceptable salt, pharmaceutical product, hydrate, isomer, N-oxide, prodrug, or polymorph of the compound according to the present invention.

The compounds of the present invention may also exist in the form of a hydrate, which means that the compound further includes a stoichiometric or non-stoichiometric amount of water attached through non-covalent intermolecular bonds.

The compounds of the present invention may exist as one or more possible tautomers, and depending on the particular conditions, some or all of the tautomers may be separated into separate and distinct molecules. It should be understood that all possible tautomers, including all additional enol and keto tautomers, and/or isomers are covered by the present invention. For example, the invention includes, but is not limited to, the following tautomers.

Tautomerization of the imidazole ring

Indazole ring tautomers

Benzimidazole ring tautomers

Pyridinoimidazole ring tautomers

The tautomers of the present invention are tautomers that are freely convertible into each other but are not inseparable mixtures. The imidazoles, thiazoles and other ring systems of the present invention may exist as tautomers. All tautomers are considered part of the invention.

The invention includes "pharmaceutically acceptable salts" of the compounds of the present invention, which can be prepared by reacting a compound of the present invention with an acid or base. Certain compounds, in particular those containing acidic or basic groups, may also be in the form of a salt, preferably a pharmaceutically acceptable salt. The term "pharmaceutically acceptable salt" refers to salts that retain the biological effectiveness and properties of free bases or free acids and are not undesirable for biological or other reasons. Salts are prepared using inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., organic acids such as acetic acid, propanoic acid, glycolic acid, grape acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine, etc. Other salts are known to those skilled in the art that can be readily adapted for use in the present invention.

Suitable pharmaceutically acceptable amine salts of the compounds of the present invention can be prepared from an inorganic acid or an organic acid. In one embodiment, examples of inorganic amine salts include bisulfates, borates, bromides, chlorides, hemisulfates, hydrobromates, hydrochlorates, 2-hydroxyethylsulfonates (hydroxyethanesulfonates), iodates, iodides, isothionates, nitrates, persulfates, phosphates, sulfates, sulfamates, sulfonylates, salts sulfonic acids (alkylsulfonates, arylsulfonates, halogenated alkylsulfonates, halogenated arylsulfonates), sulfonates and thiocyanates.

In one embodiment, examples of organic amine salts may be selected from the classes of aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and organosulfonic acids, examples of which are acetates, arginines, aspartates, ascorbates, adipates, anthranilates, algenates, alkane carboxylates, substituted alkanecarboxylates, alginates, benzenesulfonates, benzoates, bisulfates, butyrates, bicarbonates, bitartrates, citrates, camphorates, camphorsulfonates, cyclohexylsulfamates, cyclopentanepropionates, calcium edetates, camsylates, carbonates, clavulanates, cinnamates, dicarboxylates, digluconates, dodecylsulfonates, dihydro chlorides, decanoates, enantuates, ethanesulfonates, edetates, edisilates, estolates, esilates, fumarates, formates, fluorides, galacturonates, gluconates, glutamates, glycolates, glucorates, glucoheptanoates, glycerophosphates, gluceptates, glycolylarsanilates, glutarates, glutamates, heptanoates, hexanoates, hydroxymaleates, hydroxy salts carboxylic acids, hexylresorcinates, hydroxybenzoates, hydroxynaphthoates, hydrofluorates, lactates, lactobionates, laurates, malates, maleates, methylenebis(beta-hydroxynaphthoates), malonates, mandelates, mesylates, methanesulfonates, methyl bromides, methyl nitrates, methylsulfonates, monopotassium maleates, mucates, monocarboxylates, naphthalene sulfonates, 2-naphthalene sulfonates, nicotinates, nitrates, napsylates, N-methylglucamines, oxalates, octanoates, oleates, pamoates, phenylacetates, picrates, phenylbenzoates, pivalates, propionates, phthalates, phenylacetate, pectinates, phenylpropionates, palmitates, pantothenates, polygalacturates, pyruvates, quinates, salicylates , succinates, stearates, sulfanilate, subacetates, tartrates, theophylline acetates, p-toluenesulfonates (tosylates), trifluoroacetates, terephthalates, tannates, theoclates, trihaloacetates, triethiodide, tricarboxylates, undecanoates and valerates.

In one embodiment, examples of inorganic carboxylic acid or hydroxyl salts may be selected from ammonium salts, alkali metals including lithium, sodium, potassium, cesium; alkaline earth metals, including calcium, magnesium, aluminum; zinc, barium, cholines, quaternary ammonium.

In another embodiment, examples of organic carboxylic acid or hydroxyl salts may be selected from arginine salts, organic amines, including aliphatic organic amines, alicyclic organic amines, aromatic organic amines, benzathines, tert-butylamines, benethamines (N-benzylphenethylamine), dicyclohexylamines, dimethylamines , diethanolamines, ethanolamines, ethylenediamines, hydrabamines, imidazoles, lysines, methylamines, meglamines, N-methyl-D-glucamines, N,N'-dibenzylethylenediamines, nicotinamides, organic amines, ornithines, pyridines, picolines, piperazines, procaine, tris(hydroxymethyl )methylamines, triethylamines, triethanolamines, trimethylamines, tromethamines and ureas.

In one embodiment, the term “isomer” includes, but is not limited to, optical isomers and analogs, structural isomers and analogs, conformational isomers and analogs, etc.

In one embodiment, the compounds of the present invention are pure (E) isomers. In another embodiment, the compounds of the present invention are pure (Z)-isomers. In another embodiment, the compounds of the present invention are a mixture of (E) and (Z) isomers. In one embodiment, the compounds of the present invention are pure (R)-isomers. In another embodiment, the compounds of the present invention are pure (S)-isomers. In another embodiment, the compounds of the present invention are a mixture of (R) and (S) isomers.

The compounds of the present invention may also be present as a racemic mixture containing substantially equivalent amounts of stereoisomers. In another embodiment, the compounds of the present invention can be prepared or otherwise isolated using known methods that provide one stereoisomer substantially free of the corresponding other stereoisomer (ie, substantially pure). By essentially pure is meant a stereoisomer that is at least 95% pure, more preferably at least about 98%, most preferably at least about 99%.

In one embodiment, salts can be prepared using conventional methods, such as reacting the free base or free acid product with one or more equivalents of the corresponding acid or base in a solvent or medium in which the salt is insoluble, or in a solvent such as water , which is removed in vacuo or by lyophilization, or by exchanging ions of the existing salt with another ion, or using a suitable ion exchange resin.

In one embodiment, the invention provides a compound of the present invention or an N-oxide thereof.

The compounds of the present invention can also be administered as metabolites. In one embodiment, the term “metabolite” refers to any substance obtained from another substance by metabolism or a metabolic process. In another embodiment, the metabolites of the present invention include M1-M14, such as those described in Example 6 and Figures 8-11.

Thus, certain derivatives, which as such have little or no pharmacological activity, when administered into or onto the surface of the body, can be converted into compounds of the present invention having the desired activity, for example, by hydrolytic cleavage. Additional information on the use of prodrugs can be found in Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (Higuchi and Stella); and Bioreversible Carriers in Drug Design, Pergamon Press (Ed. E B Roche, American Pharmaceutical Association) (1987), the contents of each of which are incorporated herein by reference in their entirety.

Prodrugs can be obtained, for example, by replacing the corresponding functional groups contained in the compounds of the present invention with certain fragments known to those skilled in the art as pro-fragments. Examples of these prodrugs include, but are not limited to, prodrugs wherein the hydrogen atom in the alcohol function (-OH) is replaced by a C ₁ -C ₆ alkyl to form an ether; and (ii) the hydrogen atom in the secondary amino group is replaced by a C ₁ -C ₁₀ alkanoyl to form an amide.

Pharmaceutical composition

Another aspect of the present invention relates to a pharmaceutical composition containing a pharmaceutically acceptable carrier and a compound according to some aspects of the present invention. The pharmaceutical composition may contain one or more of the above compounds according to the present invention. Typically, the pharmaceutical composition of the present invention contains a compound of the present invention or a pharmaceutically acceptable salt thereof, as well as a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" refers to any suitable adjuvants, carriers, excipients or stabilizers and may be in solid or liquid form, such as tablets, capsules, powders, solutions, suspensions or emulsions.

Typically the composition contains from about 0.01 to 99 percent, preferably from about 20 to 75 percent, active compound(s) together with adjuvants, carriers and/or excipients. Although the needs of individuals may vary, determination of optimal ranges of effective amounts of each component can be accomplished within the bounds of knowledge in the art. Typical dosages range from about 0.01 to about 100 mg/kg body weight. Preferred dosages range from about 0.1 to about 100 mg/kg body weight. Most preferred dosages range from about 1 to about 100 mg/kg body weight. Those skilled in the art can also readily determine a treatment regimen using the compounds of the present invention. That is, the frequency of administration and dose size can be set using conventional determination of optimal characteristics, preferably while minimizing any side effects.

Solid unit dosage forms may be of the traditional type. The solid form may be a capsule, etc., such as a conventional gelatin-type capsule containing the compounds of the present invention and a carrier, for example, lubricants and excipients such as lactose, sucrose and corn starch. In another embodiment, the compounds are prepared in tablet form using a traditional tablet base such as lactose, sucrose or corn starch, in combination with binders such as gum arabic, corn starch or gelatin, disintegrants such as corn starch, potato starch or alginic acid, or a lubricant such as stearic acid or magnesium stearate.

Tablets, capsules, etc. may also contain a binder such as gum tragacanth, gum acacia, corn starch or gelatin; excipients such as dicalcium phosphate; leavening agent such as corn starch, potato starch, alginic acid; a lubricant such as magnesium stearate; and a sweetener such as sucrose, lactose or saccharin. If the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as a fatty oil.

Various other materials may be added as coatings or to modify the physical form of the dosage form. For example, tablets can be coated with shellac, sugar, or both. The syrup may contain, in addition to the active ingredient, sucrose as a sweetener, methyl and propyl parabens as preservatives, a coloring agent and a flavoring agent such as cherry or orange flavoring.

In the case of oral therapeutic administration, these active compounds can be administered together with excipients and used in the form of tablets, capsules, elixirs, suspensions, syrups, etc. Said compositions and preparations may contain at least 0.1% of the active compound. The percentage of compound in these compositions can of course vary and typically ranges from about 2% to about 60% of the form by weight. The amount of active compound in said therapeutically suitable compositions is such as to provide a suitable dosage. Preferred compositions of the present invention are prepared such that the oral dosage form contains from about 1 mg to 800 mg of active compound.

The active compounds of the present invention may be administered orally, for example, together with an inert diluent or digestible edible carrier, or they may be enclosed in hard or soft shell capsules, or compressed into tablets, or included directly in food.

Pharmaceutical forms suitable for injection include sterile aqueous solutions or dispersions and sterile powders for the immediate preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and have sufficient fluidity for ease of use in a syringe. It must be stable under the conditions of receipt and storage and must be protected from the contaminating action of microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersing medium containing, for example, water, ethanol, polyol (eg glycerin, propylene glycol and liquid polyethylene glycol), suitable mixtures thereof and vegetable oils.

The compounds or pharmaceutical compositions of the present invention can also be administered as injectable dosages of a solution or suspension of these materials in a physiologically acceptable diluent together with a pharmaceutical adjuvant, carrier or excipient. Said adjuvants, carriers and/or excipients include, but are not limited to, sterile liquids such as water and oils, with or without added surfactants and other pharmaceutically and physiologically acceptable components. Exemplary oils are oils of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil or mineral oil. In general, preferred liquid carriers for injectable solutions are water, saline, aqueous dextrose and similar sugar solutions and glycols such as propylene glycol or polyethylene glycol.

These active compounds can also be administered parenterally. Solutions or suspensions of these active compounds can be prepared in water mixed with a suitable surfactant such as hydroxypropylcellulose. It is also possible to obtain dispersions in glycerin, liquid polyethylene glycols and their mixtures in oils. Exemplary oils are oils of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil or mineral oil. In general, preferred liquid carriers, especially for injectable solutions, are water, saline, aqueous dextrose and similar sugar solutions and glycols such as propylene glycol or polyethylene glycol. Under standard conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

For aerosol use, the compounds of the present invention, in solution or suspension form, can be packaged in containers containing the pressurized aerosol together with suitable propellants, for example, hydrocarbon propellants such as propane, butane or isobutane, and conventional adjuvants. The materials of the present invention can also be administered in atmospheric pressure forms, such as a nebulizer or nebulizer.

In one embodiment, the compounds of the present invention are administered in combination with an anticancer agent. In one embodiment, the anticancer agent is a monoclonal antibody. In some embodiments, monoclonal antibodies are used to diagnose, monitor, or treat cancer. In one embodiment, the monoclonal antibodies target specific antigens on cancer cells. In one embodiment, the monoclonal antibody acts as a cancer cell receptor antagonist. In one embodiment, the monoclonal antibodies enhance the patient's immune response. In one embodiment, the monoclonal antibodies target cell growth factors and thereby block the growth of cancer cells. In one embodiment, the anticancer monoclonal antibodies are conjugated or linked to anticancer drugs, radioisotopes, other biological response modifiers, other toxins, or a combination thereof. In one embodiment, the anti-cancer monoclonal antibodies are conjugated or linked to a compound of the present invention, such as those described above herein.

Another aspect of the present invention relates to a method of treating cancer, comprising selecting a subject in need of cancer treatment and administering to the subject a pharmaceutical composition comprising a compound of the first aspect of the present invention and a pharmaceutically acceptable carrier under conditions that are effective for treating the cancer.

Administration of the compounds of the present invention may be administered systemically or alternatively directly to a specific site where cancer cells or precancerous cells are present. Thus, administration can be accomplished by any method that is effective in delivering compounds or pharmaceutical compositions to cancer cells or precancerous cells. Exemplary routes of administration include, but are not limited to, administration of the compounds or compositions by oral, topical, transdermal, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, intranasal, intracavitary or intravesical, intraocular, intraarterial, intralesional, or mucosal membrane application routes. such as those found in the nose, throat and bronchial tubes.

Biological activity

The compounds of the present invention are suitable for the treatment or prevention of various forms of cancer, in particular prostate cancer, breast cancer, drug-resistant breast cancer, ovarian cancer, skin cancer (for example, melanoma), drug-resistant melanoma, lung cancer, colon cancer, leukemia, kidney cancer, central nervous system cancer (for example, glioma, glioblastoma). Methods for treating these various types of cancer are substantiated in the examples given herein. In addition, given the mode of action of tubulin inhibitors, other forms of cancer are likely to be treated or prevented when the compounds or compositions of the present invention are administered to a patient. Preferred compounds of the present invention selectively destroy cancer cells, cause depletion of cancer cells, but preferably not normal cells. It is important to note that harm to normal cells is minimized, since cancer cells are susceptible to destruction when using significantly lower concentrations of the compounds of the present invention.

Thus, a further aspect of the present invention relates to a method of destroying a cancer cell, comprising: providing a compound of the present invention and then contacting the cancer cell with the compound under conditions that are effective to destroy the contacted cancer cell. In various embodiments of cancer cell destruction, the cells being destroyed may be in vivo or ex vivo (ie, in culture).

A further aspect of the present invention relates to a method of treating or preventing a cancerous condition, comprising: providing a compound of the present invention and then administering an effective amount of the compound to a patient by a method that is effective to treat or prevent the cancerous condition.

In one embodiment, the patient being treated has a precancerous condition, and administration of the compound is effective in preventing the precancerous condition from progressing to a cancerous condition. This prevention may occur by destroying the precancerous cell before or simultaneously with its further development into a cancerous state.

In another embodiment, the patient being treated has a cancerous condition, and administration of the compound is effective in reversing the cancerous condition or suppressing the progression of the cancerous condition, i.e. stopping its development or reducing the rate of development, which preferably occurs when cancer cells are destroyed regardless of their location in the patient's body, that is, cancer cells that are located in the primary site of the tumor, or cancer cells that have metastasized and created secondary tumors in the patient's body.

If the compounds or pharmaceutical compositions of the present invention are administered for the treatment or prevention of a cancerous condition, the pharmaceutical composition may also contain other therapeutic agents, or it may be used in conjunction with other treatment regimens, currently known or those hereafter developed, for the treatment various types of cancer. Examples of other therapeutic agents or treatment regimens include, but are not limited to, radiation therapy, immunotherapy, chemotherapy, surgery, and combinations thereof.

In one embodiment, the invention provides compounds and compositions, including any embodiments described herein, for use in any of the methods of the present invention. In one embodiment, a compound of the present invention or a composition containing it can be used to suppress, attenuate, enhance or stimulate a target response in a subject, as will be apparent to those skilled in the art. In another embodiment, the compositions may contain additional active ingredients that have suitable activity for the particular application for which the compound of the present invention is administered.

In one embodiment, the present invention provides a method of treating, mitigating, reducing the severity of, reducing the risk of developing or suppressing cancer, comprising administering a compound of the present invention to a subject suffering from cancer under conditions that are effective in treating cancer.

Drug resistance is the leading cause of chemotherapy failure against cancer. One of the main factors of multidrug resistance is increased expression of P-glycoprotein (P-gp). The protein is a clinically important transport protein belonging to the family of ATP-binding cassette transporters of the cell membrane. It can remove substrates, including anticancer drugs, from tumor cells through an ATP-dependent mechanism.

In one embodiment, the present invention provides methods for: a) treating, reducing, reducing the severity of, reducing the risk of, or suppressing cancer; b) treating, reducing, reducing the severity, reducing the risk of or suppressing prostate cancer; c) treating, reducing, reducing the severity of, reducing the risk of, or suppressing drug-resistant prostate cancer; d) treating, mitigating, reducing the severity, reducing the risk or suppressing breast cancer; f) treating, reducing, reducing the severity, reducing the risk of, or suppressing drug-resistant breast cancer; f) treating, reducing, reducing the severity of, reducing the risk of, or suppressing ovarian cancer; g) treating, mitigating, reducing the severity, reducing the risk of, or suppressing drug-resistant ovarian cancer; h) treating, reducing, reducing the severity of, reducing the risk of, or suppressing skin cancer; i) treating, mitigating, reducing the severity, reducing the risk or suppressing melanoma; j) treating, reducing, reducing the severity, reducing the risk of, or suppressing metastatic melanoma; k) treating, mitigating, reducing the severity, reducing the risk of, or suppressing drug-resistant melanoma; l) treating, mitigating, reducing the severity, reducing the risk of or suppressing lung cancer; m) treating, reducing, reducing the severity of, reducing the risk of, or suppressing colon cancer; n) treating, attenuating, reducing the severity, reducing the risk or suppressing glioma; o) treating, weakening, reducing the severity, reducing the risk or suppressing leukemia; p) treating, reducing, reducing the severity of, reducing the risk of, or suppressing lymphoma; q) treating, reducing, reducing the severity of, reducing the risk of, or suppressing kidney cancer; r) treating, mitigating, reducing the severity, reducing the risk of, or suppressing CNS cancer; s) treating, mitigating, reducing the severity, reducing the risk of or suppressing uterine cancer; t) treating, mitigating, reducing the severity, reducing the risk of or suppressing drug-resistant uterine cancer; u) treating a drug-resistant tumor or tumors, wherein said tumor is selected from the group consisting of prostate cancer, drug-resistant prostate cancer, breast cancer, drug-resistant breast cancer, glial tumor, ovarian cancer, cancer tumor drug-resistant ovarian cancer, skin cancer, melanoma tumor, drug-resistant melanoma tumor, lung cancer, colon cancer, leukemia tumor, lymphoma tumor, kidney cancer, central nervous system cancer, uterine cancer, drug-resistant uterine cancer stability and their combinations; comprising administering a compound of the present invention and/or a metabolite, a pharmaceutically acceptable salt, a pharmaceutical product, a tautomer. a hydrate, isomer, N-oxide, prodrug, polymorph of the specified compound or any combination thereof. In another embodiment, the subject has previously been treated with chemotherapy, radiotherapy, or biological therapy.

The compounds of the present invention are suitable for the treatment, reduction of severity, risk reduction or suppression of cancer, metastatic cancer, drug-resistant tumors, drug-resistant cancer and various forms of cancer. In a preferred embodiment, the cancer is prostate cancer, drug-resistant prostate cancer, breast cancer, drug-resistant breast cancer, ovarian cancer, drug-resistant ovarian cancer, uterine cancer, drug-resistant uterine cancer, skin cancer (e.g., melanoma) , drug-resistant melanoma, lung cancer, colon cancer, leukemia, lymphoma, head and neck cancer, pancreatic cancer, esophageal cancer, kidney cancer, or central nervous system cancer (eg, glioma, glioblastoma). Methods for treating these various types of cancer are substantiated in the examples given herein. In addition, given the mode of action of tubulin inhibitors, other forms of cancer are likely to be treated or prevented when the compounds or compositions of the present invention are administered to a patient. Preferred compounds of the present invention selectively destroy cancer cells, cause depletion of cancer cells, but preferably not normal cells. It is important to note that harm to normal cells is minimized, since cancer cells are susceptible to destruction when using significantly lower concentrations of the compounds of the present invention.

In some embodiments, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, polymorph, hydrate, isomer, N-oxide, prodrug, or any combination thereof, to treat, alleviate, reduce severity, risk reduction, or suppression of cancer in a subject. In another embodiment, the cancer is an adrenal cortical carcinoma, anal cancer, bladder cancer, a brain tumor, a brainstem tumor, breast cancer, drug-resistant breast cancer, glioma, cerebellar astrocytoma, cerebral astrocytoma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumors of the pineal gland, hypothalamic glioma, carcinoid tumor, carcinoma, cervical cancer, colon cancer, central nervous system (CNS) cancer, endometrial cancer, esophageal cancer, extrahepatic bile duct cancer, Ewing family of tumors (Pnet), extracranial embryonic cell tumor, eye cancer, intraocular melanoma, gallbladder cancer, gastric cancer, embryonal cell tumor, extragonadal gestational trophoblastic tumor, head and neck cancer, hypopharyngeal cancer, islet cell carcinoma, laryngeal cancer, leukemia, acute lymphoblastic leukemia, oral cancer , liver cancer, lung cancer, non-small cell lung cancer, small cell lymphoma, AIDS-related lymphoma, (primary) central nervous system lymphoma, cutaneous T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, malignant mesothelioma, melanoma, drug-resistant melanoma, Merkel cell carcinoma, metastatic squamous cell carcinoma, multiple myeloma, plasma cell neoplasms, mycosis fungoides, myelodysplastic syndrome, myeloproliferative disorders, nasopharyngeal cancer, neuroblastoma, oropharyngeal cancer, osteosarcoma, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, borderline tumor ovarian, drug-resistant ovarian cancer, pancreatic cancer, exocrine pancreatic cancer, islet cell carcinoma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pheochromocytoma, pituitary cancer, plasma cell neoplasms, prostate cancer, cancer drug-resistant prostate, rhabdomyosarcoma, colorectal cancer, kidney cancer, renal cell cancer, salivary gland cancer, Sezary syndrome, skin cancer, cutaneous T-cell lymphoma, skin cancer, Kaposi's sarcoma, skin cancer, melanoma, drug-resistant melanoma , small intestine cancer, soft tissue sarcoma, testicular cancer, malignant thymoma, thyroid cancer, urethral cancer, uterine cancer, drug-resistant uterine cancer, sarcoma, rare childhood cancers, vaginal cancer, vulvar cancer, Wilms tumor or any combination thereof. In another embodiment, the subject has previously been treated with chemotherapy, radiotherapy, or biological therapy.

In some embodiments, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, polymorph, hydrate, isomer, N-oxide, prodrug, or any combination thereof, to treat, alleviate, reduce severity, reduction of risk, or suppression of metastatic cancer in a subject. In another embodiment, the cancer is an adrenal cortical carcinoma, anal cancer, bladder cancer, a brain tumor, a brainstem tumor, breast cancer, drug-resistant breast cancer, glioma, cerebellar astrocytoma, cerebral astrocytoma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumors of the pineal gland, hypothalamic glioma, carcinoid tumor, carcinoma, cervical cancer, colon cancer, central nervous system (CNS) cancer, endometrial cancer, esophageal cancer, extrahepatic bile duct cancer, Ewing family of tumors (Pnet), extracranial embryonic cell tumor, eye cancer, intraocular melanoma, gallbladder cancer, gastric cancer, embryonal cell tumor, extragonadal gestational trophoblastic tumor, head and neck cancer, hypopharyngeal cancer, islet cell carcinoma, laryngeal cancer, leukemia, acute lymphoblastic leukemia, oral cancer , liver cancer, lung cancer, non-small cell lung cancer, small cell lymphoma, AIDS-related lymphoma, (primary) central nervous system lymphoma, cutaneous T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, malignant mesothelioma, melanoma, drug-resistant melanoma, Merkel cell carcinoma, metastatic squamous cell carcinoma, multiple myeloma, plasma cell neoplasms, mycosis fungoides, myelodysplastic syndrome, myeloproliferative disorders, nasopharyngeal cancer, neuroblastoma, oropharyngeal cancer, osteosarcoma, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, borderline tumor ovarian, drug-resistant ovarian cancer, pancreatic cancer, exocrine pancreatic cancer, islet cell carcinoma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pheochromocytoma, pituitary cancer, plasma cell neoplasms, prostate cancer, cancer drug-resistant prostate, rhabdomyosarcoma, colorectal cancer, kidney cancer, renal cell cancer, salivary gland cancer, Sezary syndrome, skin cancer, cutaneous T-cell lymphoma, skin cancer, Kaposi's sarcoma, skin cancer, melanoma, drug-resistant melanoma , small intestine cancer, soft tissue sarcoma, testicular cancer, malignant thymoma, thyroid cancer, urethral cancer, uterine cancer, drug-resistant uterine cancer, sarcoma, rare childhood cancers, vaginal cancer, vulvar cancer, Wilms tumor or any combination thereof.

In some embodiments, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, polymorph, hydrate, isomer, N-oxide, prodrug, or any combination thereof, to treat, alleviate, reduce the severity of, reducing the risk of, or suppressing drug-resistant or refractory cancer in a subject. In another embodiment, the cancer is an adrenal cortical carcinoma, anal cancer, bladder cancer, a brain tumor, a brainstem tumor, breast cancer, drug-resistant breast cancer, glioma, cerebellar astrocytoma, cerebral astrocytoma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumors of the pineal gland, hypothalamic glioma, carcinoid tumor, carcinoma, cervical cancer, colon cancer, central nervous system (CNS) cancer, endometrial cancer, esophageal cancer, extrahepatic bile duct cancer, Ewing family of tumors (Pnet), extracranial embryonic cell tumor, eye cancer, intraocular melanoma, gallbladder cancer, gastric cancer, embryonal cell tumor, extragonadal gestational trophoblastic tumor, head and neck cancer, hypopharyngeal cancer, islet cell carcinoma, laryngeal cancer, leukemia, acute lymphoblastic leukemia, oral cancer , liver cancer, lung cancer, non-small cell lung cancer, small cell lymphoma, AIDS-related lymphoma, (primary) central nervous system lymphoma, cutaneous T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, malignant mesothelioma, melanoma, drug-resistant melanoma, Merkel cell carcinoma, metastatic squamous cell carcinoma, multiple myeloma, plasma cell neoplasms, mycosis fungoides, myelodysplastic syndrome, myeloproliferative disorders, nasopharyngeal cancer, neuroblastoma, oropharyngeal cancer, osteosarcoma, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, borderline tumor ovarian, pancreatic cancer, exocrine pancreatic cancer, islet cell carcinoma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pheochromocytoma, pituitary cancer, plasma cell neoplasms, prostate cancer, rhabdomyosarcoma, rectal cancer, cancer kidney, renal cell carcinoma, salivary gland cancer, Sezary syndrome, skin cancer, cutaneous T-cell lymphoma, skin cancer, Kaposi's sarcoma, skin cancer, melanoma, drug-resistant melanoma, small intestinal cancer, soft tissue sarcoma, testicular cancer, malignant thymoma, thyroid cancer, urethral cancer, uterine cancer, sarcoma, rare childhood cancers, vaginal cancer, cancer of the external genitalia, Wilms tumor, or any combination thereof.

In one embodiment, the term “metastatic cancer” refers to cancer that has spread (metastasized) from its original site to another area of the body. In theory, all cancers have the potential to spread. The possibility of developing metastases depends on the complex interaction of many factors affecting cancer cells, including the type of cancer, the degree of maturity (differentiation) of tumor cells, the location and duration of cancer, as well as other factors that are not fully understood. Metastases spread in three ways—by local expansion of the tumor into surrounding tissue, through the bloodstream to distant sites, or through the lymphatic system to nearby or distant lymph nodes. Each type of cancer has a typical way of spreading. The name of the tumor is determined by the primary site (for example, breast cancer that has spread to the brain is called breast cancer that has metastasized to the brain).

In one embodiment, the term “drug-resistant cancer” refers to cancer cells that become resistant to chemotherapy. Cancer cells can become resistant to chemotherapy through a variety of mechanisms, including mutation or overexpression of the drug target, inactivation of the drug, or removal of the drug from the cell. Tumors that recur after an initial response to chemotherapy may be resistant to multiple drugs (multidrug resistant). In the traditional case of drug resistance, one or more cells from a tumor population acquire genetic changes that confer drug resistance. Accordingly, the causes of drug resistance are, among others: a) mutation (change) of some cells that are not destroyed as a result of chemotherapy, leading to drug resistance. As a result, the number of resistant cells increases and may begin to exceed the number of cells susceptible to chemotherapy; b) gene amplification. A cancer cell can make hundreds of copies of a particular gene. This gene triggers increased production of a protein that makes the use of an anticancer drug ineffective; c) removing the drug from cancer cells as soon as it enters the cell under the action of a molecule called P-glycoprotein; d) stopping the flow of the drug into cancer cells due to the cessation of the functioning of the protein that ensures the transport of the drug through the cell wall; f) recognition by cancer cells of the method of repairing DNA breaks caused by certain anticancer drugs; f) development of a drug inactivation mechanism in cancer cells. One of the main factors of multidrug resistance is the increased expression of P-glycoprotein (P-gp). The protein is a clinically important transport protein belonging to the family of ATP-binding cassette transporters of the cell membrane. It can remove substrates, including anticancer drugs, from tumor cells through an ATP-dependent mechanism. Thus, resistance to anticancer agents used in chemotherapy is the main reason for the ineffective treatment of malignant disorders, causing an increase in tumor resistance. Drug resistance is the leading cause of failure of cancer chemotherapy.

In one embodiment, the term “resistant cancer” refers to drug-resistant cancer, such as described above in this application. In another embodiment, “resistant cancer” refers to cancer cells that become resistant to any treatment, such as chemotherapy, radiotherapy, or biological therapy.

In one embodiment, the present invention provides a method of treating, reducing, reducing the severity, reducing the risk of, or suppressing cancer in a subject where the subject has previously been treated with chemotherapy, radiotherapy, or biological therapy.

In one embodiment, the term “chemotherapy” refers to a chemical treatment for cancer, such as the use of drugs that directly destroy cancer cells. These drugs are called “anti-cancer” drugs or “anti-cancer drugs”. Currently, more than 100 drugs are used in therapy to treat cancer. When treating a specific type of cancer, chemotherapy is used to control tumor growth if cure is not possible; to shrink a tumor before surgery or radiation therapy; to relieve symptoms (such as pain); and to kill microscopic cancer cells that may persist after a known tumor is removed by surgery (called adjuvant therapy). Adjuvant therapy is given to prevent the cancer from returning.

In one embodiment, the term “radiotherapy” refers to the use of high-energy X-rays and similar rays (such as electrons) to treat a disease. Many people suffering from cancer receive radiotherapy as part of their treatment. Radiotherapy can be performed as external radiotherapy, using X-rays from a source outside the body, or as internal radiotherapy, using X-rays from a source inside the body. The effect of radiotherapy is based on the destruction of cancer cells in the affected area. Although radiation therapy can also destroy normal cells, they can usually repair themselves. Radiation therapy treatment can cure some types of cancer and also reduce the chance of cancer coming back after surgery. It can be used to reduce cancer symptoms.

In one embodiment, the term “biological therapy” refers to the use of substances that are normally present in the body to destroy cancer cells. There are several types of treatment, including monoclonal antibodies, cancer suppressive agents, vaccines, and gene therapy. Biological therapy is also called immunotherapy.

In one embodiment, the present invention provides a method of treating a subject suffering from prostate cancer, metastatic prostate cancer, resistant prostate cancer or drug-resistant prostate cancer, comprising the step of administering to said subject a compound of the present invention or a metabolite thereof, a pharmaceutically acceptable salt, a pharmaceutical a product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, or a composition containing the same, in an amount that is effective to cure prostate cancer in a subject. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In one embodiment, the present invention provides a method of attenuating, reducing the severity, reducing the risk of, delaying the progression of, or suppressing prostate cancer, metastatic prostate cancer, resistant prostate cancer, or drug-resistant prostate cancer in a subject, comprising administering to the subject a compound of the present invention and/ or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph or any combination thereof or a composition containing the same. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In one embodiment, the present invention provides a method of treating a subject suffering from breast cancer, metastatic breast cancer, resistant breast cancer or drug-resistant breast cancer, comprising the step of administering to said subject a compound of the present invention or a metabolite thereof, a pharmaceutically acceptable salt, a pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof or compositions containing the same. In another embodiment, the subject is a man or a woman. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In one embodiment, the present invention provides a method of attenuating, reducing the severity, reducing the risk of, delaying the progression of, or suppressing breast cancer, metastatic breast cancer, resistant breast cancer, or drug-resistant breast cancer in a subject, comprising the step of administering to said subject a compound of the present invention or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph or any combination thereof or a composition containing the same. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of ovarian cancer, metastatic ovarian cancer, resistant ovarian cancer or drug-resistant ovarian cancer in a subject. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In one embodiment, the present invention provides a method of treating, ameliorating, reducing the severity, reducing the risk of, or suppressing melanoma, metastatic melanoma, resistant melanoma, or drug-resistant melanoma in a subject, comprising administering to the subject a compound of the present invention and/or a metabolite thereof, pharmaceutically an acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of lung cancer, metastatic lung cancer, resistant lung cancer or drug-resistant lung cancer. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of non-small cell lung cancer, metastatic small cell lung cancer, resistant small cell lung cancer or drug-resistant small cell lung cancer. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of colon cancer, metastatic colon cancer, resistant colon cancer or drug-resistant colon cancer. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of leukemia, metastatic leukemia, resistant leukemia or drug-resistant leukemia. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of glioma, metastatic glioma, resistant glioma or drug-resistant glioma. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of lymphoma, metastatic lymphoma, resistant lymphoma or drug-resistant lymphoma. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of head and neck cancer, metastatic head and neck cancer, resistant head and neck cancer or drug-resistant head and neck cancer. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of pancreatic cancer, metastatic pancreatic cancer, resistant pancreatic cancer or drug-resistant pancreatic cancer. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of esophageal cancer, metastatic esophageal cancer, resistant esophageal cancer or drug-resistant esophageal cancer. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of kidney cancer, metastatic kidney cancer, resistant kidney cancer or drug-resistant kidney cancer. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, isomer, N-oxide, prodrug, polymorph, or any combination thereof, for treating, ameliorating, reducing severity, risk reduction, delay of progression or suppression of CNS cancer, metastatic CNS cancer, resistant CNS cancer or drug-resistant CNS cancer. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In some embodiments, the present invention provides the use of a compound described herein, or a metabolite, pharmaceutically acceptable salt, pharmaceutical product, tautomer, polymorph, hydrate, isomer, N-oxide, prodrug, or any combination thereof, to treat, alleviate, reduce severity, risk reduction, or suppression of the subject's cancer(s) or drug-resistant tumors. In another embodiment, the cancer is an adrenal cortical carcinoma, anal cancer, bladder cancer, a brain tumor, a brainstem tumor, breast cancer, drug-resistant breast cancer, glioma, cerebellar astrocytoma, cerebral astrocytoma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumors of the pineal gland, hypothalamic glioma, carcinoid tumor, carcinoma, cervical cancer, colon cancer, central nervous system (CNS) cancer, endometrial cancer, esophageal cancer, extrahepatic bile duct cancer, Ewing family of tumors (Pnet), extracranial embryonic cell tumor, eye cancer, intraocular melanoma, gallbladder cancer, gastric cancer, embryonal cell tumor, extragonadal gestational trophoblastic tumor, head and neck cancer, hypopharyngeal cancer, islet cell carcinoma, laryngeal cancer, leukemia, acute lymphoblastic leukemia, oral cancer , liver cancer, lung cancer, non-small cell lung cancer, small cell lymphoma, AIDS-related lymphoma, (primary) central nervous system lymphoma, cutaneous T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, malignant mesothelioma, melanoma, drug-resistant melanoma, Merkel cell carcinoma, metastatic squamous cell carcinoma, multiple myeloma, plasma cell neoplasms, mycosis fungoides, myelodysplastic syndrome, myeloproliferative disorders, nasopharyngeal cancer, neuroblastoma, oropharyngeal cancer, osteosarcoma, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, borderline tumor ovarian, pancreatic cancer, exocrine pancreatic cancer, islet cell carcinoma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pheochromocytoma, pituitary cancer, plasma cell neoplasms, prostate cancer, rhabdomyosarcoma, rectal cancer, cancer kidney, renal cell carcinoma, salivary gland cancer, Sezary syndrome, skin cancer, cutaneous T-cell lymphoma, skin cancer, Kaposi's sarcoma, skin cancer, melanoma, drug-resistant melanoma, small intestinal cancer, soft tissue sarcoma, testicular cancer, malignant thymoma, thyroid cancer, urethral cancer, uterine cancer, sarcoma, rare childhood cancers, vaginal cancer, cancer of the external genitalia, Wilms tumor, or any combination thereof. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the tumor is a prostate cancer tumor. In another embodiment, the tumor is a multidrug-resistant (MDR) prostate cancer. In another embodiment, the tumor is an ovarian cancer. In another embodiment, the tumor is a multidrug-resistant (MDR) ovarian cancer. In another embodiment, the tumor is a uterine cancer. In another embodiment, the tumor is a multidrug-resistant (MDR) uterine cancer. In another embodiment, the tumor is a melanoma tumor. In another embodiment, the tumor is a drug-resistant melanoma tumor. In another embodiment, the tumor is a multidrug-resistant (MDR) melanoma tumor. In another embodiment, the tumor is a lung cancer tumor. In another embodiment, the tumor is a colon cancer tumor. In another embodiment, the tumor is a breast cancer. In another embodiment, the tumor is a drug-resistant breast cancer. In another embodiment, the tumor is a glial tumor. In another embodiment, the tumor is a leukemic tumor.

In one embodiment, the present invention provides a method of destroying a cancer cell, comprising: providing a compound of the present invention and contacting the cancer cell with the compound under conditions that are effective to destroy the contacted cancer cell. In various embodiments of the destruction of cancer cells, the cells to be destroyed may be in vivo or ex vivo (ie, in culture). In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the compound is compound 15. In another embodiment, the compound is compound 18. In another embodiment, the compound is compound 33. In another embodiment, the cancer is selected from the group consisting of prostate cancer, drug-resistant prostate cancer , breast cancer, drug-resistant breast cancer, ovarian cancer, drug-resistant ovarian cancer, skin cancer, melanoma, drug-resistant melanoma, lung cancer, colon cancer, leukemia, glioma, kidney cancer, central nervous system cancer, uterine cancer, cancer drug-resistant uteri and their combinations.

In one embodiment, the present invention provides a method of inhibiting, preventing or slowing the progression of tumor vascularization, comprising administering a compound of the present invention to a subject with cancer under conditions that are effective to inhibit, prevent or slow the progression of vascularization of the tumor. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In one embodiment, the present invention provides a method of inhibiting, preventing, or slowing the progression of vascularization of a metastatic tumor, comprising administering a compound of the present invention to a subject with cancer under conditions that are effective to inhibit, prevent, or slow the progression of vascularization of said tumor. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In another embodiment, the tumor is selected from the group consisting of prostate cancer, drug-resistant prostate cancer, breast cancer, drug-resistant breast cancer, glial tumor, ovarian cancer, drug-resistant ovarian cancer, skin cancer , melanoma tumor, drug-resistant melanoma tumor, lung cancer, colon cancer, lymphoma tumor, kidney cancer, central nervous system cancer, uterine cancer, drug-resistant uterine cancer, and combinations thereof.

A further aspect of the present invention relates to a method of treating or preventing a cancerous condition, comprising: providing a compound of the present invention and then administering an effective amount of the compound to a patient by a method that is effective for treating or preventing the cancerous condition.

In one embodiment, the patient being treated has a precancerous condition, and administration of the compound is effective in preventing the precancerous condition from progressing to a cancerous condition. This prevention may occur by destroying the precancerous cell before or simultaneously with its further development into a cancerous state.

In another embodiment, the patient being treated has a cancerous condition, and administration of the compound is effective in reversing the cancerous condition or suppressing the progression of the cancerous condition, i.e. stopping its development or reducing the rate of development, which preferably occurs when cancer cells are destroyed regardless of their location in the patient's body, that is, cancer cells that are located in the primary site of the tumor, or cancer cells that have metastasized and created secondary tumors in the patient's body.

As used herein, a subject or patient refers to any mammalian patient, including, without limitation, humans and other primates, dogs, cats, horses, cows, sheep, pigs, rats, mice and other rodents. In one embodiment, the subject is a human. In one embodiment, the subject is a man. In another embodiment, the subject is a woman. In some embodiments, the methods described herein may be suitable for treating men or women.

Administration of the compounds of the present invention may be administered systemically or alternatively directly to a specific site where cancer cells or precancerous cells are present. Thus, administration can be accomplished by any method that is effective for delivering compounds or pharmaceutical compositions to cancer cells or precancerous cells. Exemplary routes of administration include, but are not limited to, administration of the compounds or compositions by oral, topical, transdermal, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, intranasal instillation, intracavitary or intravesical instillation, intraocular, intraarterial, intralesional, or mucosal membrane application. such as those found in the nose, throat and bronchial tubes.

The compounds of the present invention are suitable for the treatment or prevention of various forms of cancer, in particular prostate cancer, drug-resistant prostate cancer, breast cancer, drug-resistant breast cancer, ovarian cancer, drug-resistant ovarian cancer, skin cancer (e.g. melanoma), drug-resistant melanoma, lung cancer, colon cancer, glioma, leukemia, lymphoma, kidney cancer, uterine cancer, drug-resistant uterine cancer and CNS cancer (eg, glioma, glioblastoma). In addition, given the mode of action of tubulin inhibitors, other forms of cancer are likely to be treated or prevented when the compounds or compositions of the present invention are administered to a patient. Preferred compounds of the present invention selectively destroy cancer cells, cause depletion of cancer cells, but preferably not normal cells. It is important to note that harm to normal cells is minimized, since cancer cells are susceptible to destruction when using significantly lower concentrations of the compounds of the present invention.

The compounds of the present invention are suitable for treating, reducing the severity of, reducing the risk of or suppressing cancer, metastatic cancer, resistant cancer or drug-resistant cancer. In another embodiment, the cancer is prostate cancer, breast cancer, drug-resistant breast cancer, ovarian cancer, skin cancer (eg, melanoma), drug-resistant melanoma, lung cancer, colon cancer, glioma, leukemia, lymphomas, head cancer and neck, pancreatic, esophageal, kidney cancer, uterine cancer or central nervous system cancer or combinations thereof. Methods for treating these various types of cancer are substantiated in the examples given herein. In addition, given the mode of action of tubulin inhibitors, other forms of cancer are likely to be treated or prevented when the compounds or compositions of the present invention are administered to a patient. Preferred compounds of the present invention selectively destroy cancer cells, cause depletion of cancer cells, but preferably not normal cells. It is important to note that harm to normal cells is minimized, since cancer cells are susceptible to destruction when using significantly lower concentrations of the compounds of the present invention. In one embodiment, the compound is a compound of formula I-XV. In another embodiment, the connection is connection 4. In one embodiment, the connection is connection 5. In another embodiment, the connection is connection 6. In another embodiment, the connection is connection 7. In another embodiment, the connection is connection 14 In another embodiment, the connection is connection 15. In another embodiment, the connection is connection 18. In another embodiment, the connection is connection 33.

In one embodiment, the compound is administered in combination with an anticancer agent by administering the compounds described herein alone or in combination with other agents. In one embodiment, the administration of the compound is carried out in combination with other cancer therapy (such as radiotherapy, chemotherapy, biological therapy).

If the compounds or pharmaceutical compositions of the present invention are administered to treat, alleviate, reduce the severity, reduce the risk of or suppress a cancerous condition, the pharmaceutical composition may also contain other therapeutic agents, or it may be used in conjunction with other treatment regimens currently known or those , which will be developed subsequently, for the treatment of various types of cancer. Examples of other therapeutic agents or treatment regimens include, but are not limited to, radiation therapy, immunotherapy, chemotherapy, surgery, and combinations thereof.

The following examples are provided to more fully illustrate embodiments of the invention. However, they should not be construed as limiting in any way the broad scope of the invention.

Examples

Example 1

Synthesis of compounds of formula I (Figure 2)

Materials. All reagents were purchased from Sigma-Aldrich Chemical Co., Fisher Scientific (Pittsburgh, PA, USA), AK Scientific (Mountain View, CA, USA), Oakwood Products (West Columbia, SC, USA), etc. and used without further purification. Moisture-sensitive interactions were carried out in an argon atmosphere. Typical thin-layer chromatography (TLC) was performed on aluminum-backed Uniplates (Analtech, Newark, DE, USA). Melting point was measured using a Fisher-Johns melting point meter (uncorrected). NMR spectra were obtained on a Bruker AX 300 spectrometer (Billerica, MA, USA) or a Varian Inova-500 spectrometer. Chemical shifts are expressed in parts per million (ppm) relative to TMS in CDCl ₃ . Mass spectrometry data were acquired on a Bruker ESQUIRE electron sputter/ion trap device in positive and negative ion detection mode. Elemental analysis was carried out at Atlantic Microlab Inc., (Norcross, GA, USA). Unless specifically stated otherwise, all compounds tested were >95% pure as determined by combustion analysis.

General method for preparing compounds 3-7 (Figure 2): Various aldehydes, 3-bromobenzene-1,2-diamine (3 mmol), p-toluenesulfonic acid (0.3 mmol) and 15 ml EtOH were refluxed for 24 hours in an argon atmosphere. The solvent was removed, 25 ml of water was added and the mixture was extracted with EtOAc (3x50 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography to give the desired 4-bromo-2-substituent-1H-benzo[d]imidazole.

Corresponding bromides obtained in the previous step (1 eq.), 3,4,5-trimethoxyphenylboronic acid (1 eq.), sodium carbonate solution (2 eq.) in THF (3 ml)/water (0.3 ml ) and tetrakis(triphenylphosphine)palladium (0.1 equiv.) were refluxed overnight. After adding water, the reaction mixture was extracted with ethyl acetate. The organic layer was dried over MgSO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography to obtain the target benzimidazole compounds with a fused “D” ring or imidazo[4,5-c]pyridine compound 7 (Figure 2).

2-phenyl-4-(3,4,5-trimethoxyphenyl)-1H-benzo[d]imidazole (3). ^1H NMR ( _CDCl3 ): 3.78, 3.93 (s, s, 6H), 3.91, 3.98 (s, s, 3H), 6.10, 6.82 (s, s, 2H), 7.29-8.08 (m, 8H), 9.70, 9.84 (s, br, 1H). MS (ESI) m/z 359.1 [MH] ^- , 361.4 [M+H] ⁺ . Analysis (C ₂₂ H ₂₀ N ₂ O ₃ ) C, H, N.

2-(1H-indol-3-yl)-4-(3,4,5-trimethoxyphenyl)-1H-benzo[d]imidazole (4). ^1H NMR (DMSO- _d6 ): 3.76, 3.97 (s, s, 3H), 3.90, 3.97 (s, s, 6H), 6.96-7.62 (m, H), 7.62, 7.72 (s, s, 2H), 8.16, 8.42 (s, lat, 1H), 8.58, 8.66 (d, d, 1H), 11 .57, 11.64 (s, s, 1H), 12.16, 12.60 (s, s, 1H). MS (ESI) m/z 398.1 [MH] ^- , 400.3 [M+H] ⁺ . Analysis (C ₂₄ H ₂₁ N ₃ O ₃ ) C, H, N.

2-(1H-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-benzo[d]imidazole (5). ^1H NMR (DMSO- _d6 ): 3.73, 3.87 (s, s, 6H), 3.75, 3.92 (s, s, 3H), 5.70, 5.75 (s, s, 2H), 6.32, 6.49 (s, s, 1H), 6.54, 6.85 (d, d, 1H), 7.22-7.65 (m, 4H), 8, 01, 8.42 (lat, s, 2H), 11.09, 11.37 (s, s, 1H), 12.36, 12.84 (s, s, 1H). MS (ESI) m/z 398.1 [MH] ^- , 400.1 [M+H] ⁺ . Analysis (C ₂₄ H ₂₁ N ₃ O ₃ ) C, H, N.

3-(4-(3,4,5-trimethoxyphenyl)-1H-benzo[d]imidazol-2-yl)-1H-indazole (6). ^1H NMR (DMSO- _d6 ): 3.76 (s, 3H), 3.95 (s, 6H), 6.65 (s, 1H), 6.93 (t, 1H), 7.34- 7.34 (m, 2H), 7.45-7.57 (m, 4H), 7.63 (s, 2H), 7.70 (d, 1H), 8.75 (d, 1H), 12 .96 (s, 1H), 13.77 (s, 1H). MS (ESI) m/z 399.1 [MH] ^- , 401.3 [M+H] ⁺ . Analysis (C ₂₃ H ₂₀ N ₄ O ₃ ) C, H, N.

2-(1H-indol-5-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine (7). ^1H NMR (DMSO- _d6 ): 3.77 (s, 3H), 3.96 (s, 6H), 6.61 (s, 1H), 7.46-7.48 (m, 2H), 7.59 (d, 1H), 8.08 (dd, 1H), 8.36 (d, 1H), 8.41 (s, 2H), 8.49 (s, 1H), 11.44 (s , 1H), 13.26 (s, 1H). MS (ESI) m/z 399.0 [MH] ^- , 401.3 [M+H] ⁺ . Analysis (C ₂₃ H ₂₀ N ₄ O ₃ ) C, H, N.

Example 2

Synthesis of compounds of formula XI (Figure 3)

(4-hydroxy-3,5-dimethoxyphenyl)(2-phenylthiazol-4-yl)methanone (11). Intermediates 8-10 were prepared from benzonitrile and cysteine according to the method described in Lu, Y.; Li, C. M.; Wang, Z.; Ross, C. R., 2nd; Chen, J.; Dalton, J.T.; Li, W.; Miller, D. D. Discovery of 4-substituted methoxybenzoyl-aryl-thiazole as novel anticancer agents: synthesis, biological evaluation, and structure-activity relationships. J Med Chem 2009, 52(6), 1701-11. Compound 10 [template compound "SMART" (4-substituted methoxybenzoyl-aryl-thiazole)] (500 mg, 1.4 mmol) was dissolved in CH ₂ Cl ₂ (50 ml) at RT under a protective atmosphere of argon. Anhydrous AlCl ₃ (374 mg, 2.8 mmol) was added and the reaction mixture was stirred for 12 hours. The reaction was quenched with H ₂ O (30 ml), the organic phase was separated and the aqueous phase was extracted with CH ₂ Cl ₂ (2x20 ml). The combined organic phases were washed with brine, dried over MgSO ₄ , filtered and concentrated to dryness under reduced pressure. Purification by flash column chromatography using hexanes-EtOAc provided 11 (410 mg, 85.9% yield). ^1H NMR ( _CDCl3 ): 4.00 (s, 6H), 6.02 (s, 1H), 7.47-7.48 (m, 3H), 7.91 (s, 2H), 8. 01-8.03 (m, 2H), 8.27 (s, 1H). MS (ESI) m/z 339.9 [MH] ^- , 364.1 [M+Na] ⁺ . Analysis (C ₁₈ H ₁₅ NO ₄ S) C, H, N.

2,6-dimethoxy-4-(2-phenylthiazol-4-carbonyl)phenyl-2-chloroacetate (12). At 0°C, 2-chloroacetyl chloride (100 mg, 0.9 mmol) was added to a solution of 11 (100 mg, 0.29 mmol) in CH ₂ Cl ₂ (30 ml). Triethylamine (44 mg, 0.44 mmol) was then added to the mixture and stirred until the starting material disappeared in TLC analysis. The reaction was quenched with H ₂ O (10 ml), the organic phase was separated and the aqueous phase was extracted with CH ₂ Cl ₂ (2x10 ml). The combined organic phases were washed with brine, dried over MgSO ₄ , filtered and concentrated to dryness under reduced pressure. Purification by flash column chromatography using hexanes-EtOAc provided 12 (99 mg, 81.7% yield). Melting _point 147-148°C. ^1H NMR ( _CDCl3 ): 3.92 (s, 6H), 4.42 (s, 2H), 7.47-7.49 (m, 3H), 7.82 (s, 2H), 8. 00-8.02 (m, 2H), 8.32 (s, 1H). MS (IER) m/z 418.1 [MH] ^- . Analysis (C ₂₀ H ₁₆ ClNO ₅ S) C, H, N.

2,6-dimethoxy-4-(2-phenylthiazol-4-carbonyl)phenyl-2,2,2-trifluoroacetate (13). At 0°C, trifluoroacetic anhydride (189 mg, 0.9 mmol) was added to a solution of 11 (100 mg, 0.29 mmol) in CH ₂ Cl ₂ (10 ml). DMAP (54 mg, 0.44 mmol) was then added to the mixture and stirred at RT until the starting material disappeared in TLC analysis. The reaction was quenched with H ₂ O (10 ml), the organic phase was separated and the aqueous phase was extracted with CH ₂ Cl ₂ (2x10 ml). The combined organic phases were washed with brine, dried over MgSO ₄ , filtered and concentrated to dryness under reduced pressure. Purification by flash column chromatography using hexanes-EtOAc provided 13 (89 mg, 70.2% yield). Melting _point 151-153°C. ^1H NMR ( _CDCl3 ): 3.94 (s, 6H), 7.48-7.49 (m, 3H), 7.84 (s, 2H), 8.00-8.02 (m, 2H ), 8.34 (s, 1H). MS (ESI) m/z 438.1 [M+H] ⁺ . Analysis (C ₂₀ H ₁₆₄ F ₃ NO ₅ S) C, H, N.

(4-(benzyloxy)-3,5-dimethoxyphenyl)(2-phenylthiazol-4-yl)methanone (14). Under argon, potassium carbonate (49 mg, 0.352 mmol) and benzyl bromide (33 mg, 0.194 mmol) were added to a solution of 11 (60 mg, 0.176 mmol) in 10 ml of dry DMF. The mixture was stirred for 1 hour at 100°C, then transferred to water (10 ml). Compound 14 was extracted with EtOAc, washed with distilled water, dried over magnesium sulfate and concentrated in vacuo on a rotary evaporator. The crude oily product was purified by flash column chromatography to give white solid 14 (51 mg). Yield = 62.7%. Melting _point 119-120°C. ^1H NMR ( _CDCl3 ): 3.92 (s, 6H), 5.15 (s, 2H), 7.29-7.37 (m, 3H), 7.48-7.51 (m, 5H ), 7.79 (s, 2H), 8.01-8.02 (m, 2H), 8.28 (s, 1H). MS (ESI) m/z 432.1 [M+H] ⁺ . Analysis (C ₂₅ H ₂₁ NO ₄ S) C, H, N.

(3,5-dimethoxy-4-(methoxymethoxy)phenyl)(2-phenylthiazol-4-yl)methanone (15). At 0°C, MOMCl (27 mg, 0.33 mmol) was added to a solution of 11 (75 mg, 0.22 mmol) in CH ₂ Cl ₂ (10 ml). Hunig's base (57 mg, 0.44 mmol) was then added to the mixture and stirred at RT until the starting material disappeared from TLC analysis. The reaction was quenched with H ₂ O (10 ml), the organic phase was separated and the aqueous phase was extracted with CH ₂ Cl ₂ (2x10 ml). The combined organic phases were washed with brine, dried over MgSO ₄ , filtered and concentrated to dryness under reduced pressure. Purification by flash column chromatography using hexanes-EtOAc provided 15 (83 mg, 98.0% yield) as yellow crystals. Melting _point 103-104°C. ^1H NMR ( _CDCl3 ): 3.62 (s, 3H), 3.95 (s, 6H), 5.26 (s, 2H), 7.47-7.49 (m, 3H), 7. 80 (s, 2H), 8.01-8.03 (m, 2H), 8.28 (s, 1H). MS (ESI) m/z 408.1 [M+Na] ⁺ . Analysis (C ₂₀ H ₁₉ NO ₅ S) C, H, N.

2-(2-(2,6-dimethoxy-4-(2-phenylthiazol-4-carbonyl)phenoxy)ethyl)-isoindoline-1,3-dione (17). K ₂ CO ₃ ( 97 mg, 0.7 mmol) and stirred the reaction mixture at 120°C overnight. The reaction was then quenched with water and the mixture was extracted with ethyl acetate. Concentrate the organic layer and further purify by column chromatography to obtain 132 mg of pure target product 17. Yield = 43.5%. Melting _point 148-150°C. ¹ H NMR (CDCl ₃ ) δ 3.71 (s, 6H), 4.14 (t, 2H, J=5.5 Hz), 4.41 (t, 2H, J=5.5 Hz), 7 .49-7.51 (m, 3H), 7.70 (s, 2H), 7.75 (q, 2H, J=3.0 Hz), 7.91 (q, 2H, J=3.0 Hz), 8.01-8.03 (m, 2H), 8.27 (s, 1H). MS (ESI) m/z 537.1 [M+Na] ⁺ . Analysis (C ₂₈ H ₂₂ N ₂ O ₆ S) C, H, N.

(4-(2-aminoethyl)-3,5-dimethoxyphenyl)(2-phenylthiazol-4-yl)methanone (18). Cs ₂ CO ₃ (46 mg, 0 .2 mmol) and stirred the reaction mixture for 3 days at RT until TLC analysis indicated completion of the reaction. The reaction was then quenched with ice water and the mixture was extracted with ethyl acetate. Concentrate the organic layer and further purify by column chromatography to obtain 22 mg of pure title product tert-butyl-(2-(2,6-dimethoxy-4-(2-phenylthiazol-4-carbonyl)phenoxy)ethyl)carbamate 16. Yield = 65.1%. MS (ESI) m/z 483.9 [MH] ^- , 485.1 [M+H] ⁺ . Boc-protected compound 16 was added to a solution of HCl in dioxane (4M) and stirred overnight. The precipitate was collected and washed with diethyl ether to give the HCl salt of compound 18. ^1H NMR (acetone- _d6 ): 3.09-3.13 (q, 2H, J=5.5 Hz), 3.79 (br., 2H), 3.90 (s, 6H), 4.17 (t, 2H, J=5.5 Hz), 7.55-7.58 (m, 3H), 7.66 (s, 2H), 8.02-8.04 (m, 2H), 8.68 (s, 1H). MS (ESI) m/z 385.1 [M+H] ⁺ . Analysis (C ₂₀ H ₂₀ N ₂ O ₃ S) C, H, N.

Example 3

Synthesis of phenyl-amino-thiazole/imidazole compounds (Figure 4)

N-phenyl-1H-imidazol-2-amine (21). At 0°C, BrCN (2.11 g, 20 mmol) was added in small portions to a solution of aminoacetaldehyde diethyl acetal (2.66 g, 20 mmol) in diethyl ether/hexane (20 ml, 1:1). The reaction mixture was stirred at RT overnight. The solid was removed by filtration and washed with diethyl ether. The combined filtrate was concentrated. Purification by flash column chromatography (silica gel, eluted with a gradient of dichloromethane followed by 5% methanol in dichloromethane) gave N-(2,2-diethoxyethyl)carbodiimide 19. ¹ H NMR, 500 MHz (CDCl ₃ ): 1.23 (t, 6H, J=7.0 Hz), 3.16 (t, 2H, J=6.0 Hz), 3.56 (dt, 2H), 3.64 (lat, s, 1H), 3.73 (dt, 2H), 4.58 (t, J=5.0 Hz, 1H). MS (ESI) m/z 156.8 [MH] ^- , 180.9 [M+Na] ⁺ . Aniline (1.66 g, 17.8 mmol) was dissolved in ethanol (50 ml) and a solution of 19 (2.82 g, 17.8 mmol) in 5 ml diethyl ether was added dropwise. Methanesulfonic acid (1.71 g, 17.8 mmol) was then added and the mixture was refluxed for 24 hours. The reaction mixture was poured into NaOH (0.5M) and extracted with CH ₂ Cl ₂ . Drying the MgSO ₄ and concentrating in vacuo afforded the product, which was purified by flash chromatography to give guanidine intermediate 20 (3.3 g, 73.8%). Guanidine (3 g, 12 mmol) was dissolved in HCl (5 ml, 6 M) at 0°C and then stirred for 2 hours. After consumption of the starting material, NaOH (25%) was added until a precipitate formed (pH 14). The resulting mixture was stirred for 30 minutes. The reaction mixture was then poured into NaOH (0.5M), extracted with CH ₂ Cl ₂ , dried and concentrated. Purification by flash chromatography gave 21 (1.16 g, 61%). ^1H NMR (DMSO- _d6 ): 6.68 (s, 2H), 6.75 (m, 1H), 7.17 (m, 2H), 7.34 (m, 2H), 8.58 ( s, 1H). MS (ESI) m/z 157.6 [MH] ^- , 160.0 [M + H] ⁺ .

3,4,5-trimethoxy-N-phenyl-N-(1-(phenylsulfonyl)-1H-imidazol-2-yl)benzamide (22). Benzenesulfonyl chloride (441 mg, 2.5 mmol) and triethylamine (252 _{mg ,} 2 .5 mmol). The reaction mixture was stirred overnight at room temperature. The reaction was extinguished by us. NH ₄ Cl and extracted with CH ₂ Cl ₂ mixture. Drying the MgSO ₄ and concentrating in vacuo afforded the product, which was purified by flash chromatography to give the benzenesulfonyl-protected intermediate (79 mg, 72%). The resulting intermediate was dissolved in THF and cooled to -78°C before t-BuLi (1.7M) was added under Ar. After stirring for another hour, 3,4,5-trimethoxybenzoyl chloride (47 mg, 0.26 mmol) was added and stirred overnight. The reaction mixture was poured into NH ₄ Cl (sat) and extracted with ethyl acetate. Drying the MgSO ₄ and concentrating in vacuo gave the crude product, which was purified by flash chromatography to give 22 (35%). ^1H NMR ( _CDCl3 ): 3.78 (s, 6H), 3.87 (s, 3H), 6.91 (s, 2H), 6.97 (s, 1H), 7.18 (m, 2H), 7.20 (d, 1H), 7.25 (m, 2H), 7.38 (m, 2H), 7.40 (d, 1H), 7.54 (br, 1H), 7 .59 (t, 2H). MS (ESI) m/z 491.9 [MH] ^- , 516.1 [M + Na] ⁺ .

N-(1H-imidazol-2-yl)-3,4,5-trimethoxy-N-phenylbenzamide (23). Boc ₂ O (2.68 g, 12.3 mmol) was added to a solution of N-phenyl-1H-imidazol-2-amine 21 (900 mg, 5.66 mmol) in dioxane and water (30 ml, 3:1) and NaOH (0.6 g, 15 mmol) and stirred for 4 hours. Concentrate the mixture in vacuo and purify the residue by flash chromatography to give the Boc-protected intermediate. The resulting intermediate (130 mg, 0.502 mmol) was dissolved in THF and cooled to -78°C before t-BuLi (0.65 mL, 1.7 M, 1.1 mmol) was added under Ar. After stirring for an hour, 3,4,5-trimethoxybenzoyl chloride (116 mg, 0.502 mmol) was added and stirred overnight. The reaction mixture was poured into NH ₄ Cl (sat) and extracted with ethyl acetate. Drying the MgSO ₄ and concentrating in vacuo gave the crude product, which was purified by flash chromatography to give 23 (35%). ^1H NMR ( _CDCl3 ): 3.65 (s, 6H), 3.79 (s, 3H), 6.56 (s, 2H), 6.90 (m, 2H), 7.27-7, 39 (m, 5H), 11.17 (broad, 1H). MS (ESI) m/z 351.8 [MH] ^- , 376.3 [M + Na] ⁺ .

N-phenyl-4-trityl-1H-imidazol-2-amine (24) and N-phenyl-1-trityl-1H-imidazol-2-amine (25). (Chloromethantriyl)tribenzene (5 eq.) was added to a solution of N-phenyl-1H-imidazol-2-amine 21 (159 mg, 10 mmol) in triethylamine and CH ₂ Cl ₂ , which was stirred in an inert atmosphere at 0°C. The solution was allowed to warm to RT and stirred until reaction was complete according to TLC. The reaction mixture was then concentrated in vacuo, quenched with saturated aqueous sodium bicarbonate and extracted with ethyl acetate, then dried over magnesium sulfate and concentrated in vacuo. The resulting residue was purified by flash chromatography to give two different protected products. 24: ^1H NMR (DMSO- _d6 ): 6.0 (s, 1H), 6.75 (m, 1H), 7.29-7.62 (m, 19H), 8.65 (s, 1H ), 10.62 (s, 1H). MS (ESI) m/z 399.9 [MH] ^- , 403.1 [M+H] ⁺ . 25: ^1H NMR (DMSO- _d6 ): 6.08 (s, 1H), 6.41 (s, 1H), 6.85 (s, 1H), 7.13-7.52 (m, 20H ), 8.65 (s, 1H), 10.62 (s, 1H). MS (ESI) m/z 399.8 [MH] ^- , 402.8 [M+H] ⁺ .

(2-(phenylamino)-1-trityl-1H-imidazol-4-yl)(3,4,5-trimethoxyphenyl)-methanone (26). t-BuLi (0. 34 ml, 1.7 M, 0.58 mmol) and trimethoxybenzoyl chloride (66.5 mg, 0.289 mmol). The reaction mixture was left to react overnight, then the reaction was quenched with NH ₄ Cl (sat) and extracted with ethyl acetate. Drying the MgSO ₄ and concentrating in vacuo provided the crude product, which was purified by flash chromatography to give 26 (75 mg, 43.7%). ^1H NMR (DMSO- _d6 ): 3.71 (s, 3H), 3.78 (s, 6H), 5.87 (s, 1H), 6.94 (s, 2H), 7.18- 7.58 (m, 21H). MS (ESI) m/z 594.2 [MH] ^- , 596.3 [M+H] ⁺ .

(2-(phenylamino)-1H-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (27). (2-(phenylamino)-1-trityl-1H-imidazol-4-yl)(3,4,5-trimethoxyphenyl)methanone was dissolved in a solution of HCl in diethyl ether (2M) and stirred overnight. Then a saturated NaHCO ₃ solution was added and the reaction mixture was extracted three times with diethyl ether. The combined organic layers were dried (sodium sulfate), filtered and concentrated in vacuo. The residue was purified by flash chromatography to give pure ^27.1H NMR (DMSO- _d6 ): 3.73 (s, 3H), 3.82 (s, 6H), 6.62 (s, 2H), 7. 02 (s, 2H), 7.33 (d, 2H), 7.43-7.51 (m, 3H), 7.54 (br, 1H). MS (ESI) m/z 352.1 [MH] ^- , 354.3 [M+H] ⁺ . Analysis (C ₁₉ H ₁₉ N ₃ O ₄ ) C, H, N.

N-((1H-indol-5-yl)carbamothioyl)benzamide (28). A mixture of 5-nitro-1H-indole (11 g, 67.9 mmol) and Pd/C (5%; 1 g) dissolved in ethanol (50 ml) was hydrogenated for 3 hours at 40 psi (280 kPa). The reaction mixture was filtered and excess ethanol was evaporated under reduced pressure. The solid was recrystallized from hexane to give the pure 5-aminoindole compound. Yield: 92.5%. ^1H NMR (500 MHz, CDCl ₃ ): δ 3.50 (s, 2H), 6.37 (s, 1H), 6.67 (dd, 1H), 6.95 (s, 1H), 7. 13 (s, 1H), 7.20 (d, 1H), 7.96 (br, 1H). MS (ESI) m/z 133.0 (M+H) ⁺ . React a solution of 5-aminoindole (8 g, 60.6 mmol) in acetone (150 mL) with benzoyl isothiocyanate (9.88 g, 60 mmol) at RT for approximately 4 hours until TLC analysis indicated completion reaction, compound 28 was obtained. ^1H NMR (300 MHz, CDCl ₃ ): δ 6.61 (br, 1H), 7.26-7.28 (d, 1H), 7.38-7.45 (m , 2H), 7.54-7.59 (m, 2H), 7.65-7.70 (m, 1H), 7.91-7.94 (m, 2H), 7.98 (s, 1H ), 8.27 (s, br, 1H), 9.12 (s, 1H), 12.51 (s, 1H). MS (ESI) m/z 318.1 [M + Na] ⁺ .

2-(1H-indol-5-ylamino)-N-methoxy-N-methylthiazole-4-carboxamide (32). The resulting compound 28 was filtered off as a solid and treated with 2N. NaOH in THF (120 ml). The mixture was refluxed for approximately 6 hours and allowed to warm to RT. The solvent was evaporated in vacuum. The residue was diluted with water (20 ml) and neutralized to pH 7 with 1N. HCl. The resulting solid was filtered and dried in vacuo to give 5-indolylthiourea (29). Compound 29 (0.01 mol) and ethyl bromopyruvate (0.011 mol) were dissolved in 3 ml ethanol and maintained at reflux temperature for 2 hours. Cool the reaction mixture, collect crystalline ethyl 2-(1H-indol-5-ylamino)thiazol-4-carboxylate (30) by filtration and wash with ethanol. Boiling a mixture of ethyl esters with NaOH-ethanol solution at the reverse condensation temperature gave 2-(1H-indol-5-ylamino)thiazol-4-carboxylic acid (31), which was used directly in the next steps. HCl salt HNCH 3 OCH 3 (2.6 mmol) was added to a mixture of crude acid (2.5 mmol), HBTU (2.6 _mmol ) and NMM (5.3 mmol) in CH _{2 Cl 2} (30 ml) and continued stir at RT overnight. The reaction mixture was diluted with CH ₂ Cl ₂ (20 ml) and washed successively with water, sat. NaHCO ₃ , saline and dried over MgSO ₄ . The solvent was removed under reduced pressure to give the crude product, which was purified by column chromatography to give pure Weinreb amide 2-(1H-indol-5-ylamino)-N-methoxy-N-methylthiazole-4-carboxamide (32) (total yield 45 .6% for 5 stages). ^1H NMR ( _CDCl3 ): 3.42 (s, 3H), 3.77 (s, 3H), 6.54 (m, 1H), 7.26 (m, 1H), 7.29 (m, 2H), 7.40 (d, 2H), 7.61 (m, 1H), 8.30 (br, 1H). MS (ESI) m/z 303.0 [M+H] ⁺ .

(2-((1H-indol-5-yl)amino)thiazol-4-yl)(3,4,5-trimethoxyphenyl)methanone (33). At -78°C, n-BuLi in hexane (2.5N, 2.4 ml, 6 mmol) was added to a solution of 5-bromo-1,2,3-trimethoxybenzene (1.235 g, 5.0 mmol) in 30 ml THF ) under a protective atmosphere of Ar and stirred for 10 minutes. Weinreb amide 32 (1 mmol) in 10 ml of THF was added to the lithium-containing reagent and left to stir at RT for 2 hours. The reaction was extinguished by us. NH ₄ Cl, the mixture was extracted with diethyl ether, and dried with MgSO ₄ . The solvent was removed under reduced pressure to give the crude product, which was purified by column chromatography to give pure 33 (51.7% yield). ^1H NMR (300 MHz, CDCl ₃ ) δ 3.89 (s, 6H), 3.93 (s, 3H), 6.55 (m, 1H), 7.15-7.12 (m, 1H) , 7.28-7.26 (m, 1H), 7.36 (s, 1H), 7.39 (s, 1H), 7.46 (s, 2H), 7.68 (d, 1H), 8.29 (broad, 1H). MS (ESI) m/z 432.1 (M + Na) ⁺ , 408.0 (M - H) ^- . Analysis (C ₂₃ H ₁₉ N ₃ O ₄ S) C, H, N.

Example 4

Molecular modeling of compounds according to the present invention

Molecular modeling. Molecular modeling studies were performed using published crystal structures of the α,β-tubulin dimer complex with DAMA-colchicine (Protein Data Bank code 1SA0). Modeling studies used the Schrodinger Molecular Modeling Kit (Schrodinger Inc., Portland, OR) using procedures similar to those previously described by Chen, J. et al. Bioorg Med Chem 2011, 19(16), 4782-95; Slominski, A. T. et al. The Journal of Steroid Biochemistry and Molecular Biology 2013, 137, 107-23; Xiao, M et al. J Med Chem 2013, 56(8), 3318-29; Chen, J. et al. J Med Chem 2012, 55(16), 7285-9. Briefly, structures of protein-ligand complexes were obtained using the Protein Preparation module and active ligand binding sites were determined from data for the natural ligand. The structures of the natural DAMA-colchicine ligand and the proposed tubulin inhibitors described in this study were generated and prepared for docking using the Ligprep module (Ligand Preparation), followed by docking in 1SA0. The Glidescore docking index obtained from this modeling method is the estimated binding energy (kcal/mol) of ligand binding to the tubulin dimer. A lower (more negative) value indicates more favorable binding between the ligand and the receptor. Data analysis was performed using the Maestro interface included in the software.

Compounds 4 and 7 were docked at the colchicine binding site of tubulin (Figure 6, PDB code: 1SA0) and were shown to have a very similar binding arrangement, which overlapped quite well with the natural ligand. As expected, the trimethoxy moiety in 4 or 7 occupied the trimethoxy moiety pocket of the natural ligand (DAMA-colchicine), but some shift in its location was observed. As a result of this slight displacement, the oxygen atoms of the two methoxy groups in 4 and 7 were located in the vicinity of Cys-241 in the β-subunit and provided the formation of two hydrogen bonds (dashed lines). The NH in the imidazole moiety in 4 or 7 formed another hydrogen bond with Thr-179 in the α subunit, as shown in Figure 6. It is interesting to note that by forming a planar structure in the middle part of 4 or 7, the 5-indolyl moiety changed orientation so manner to reach GTP in the α subunit. The Glidescore docking indices for 4 (-8.58) and 7 (-8.10) were comparable to the natural ligand, DAMA-colchicine (-9.26), according to these model calculations, suggesting that they have comparable activity in binding of tubulin.

Example 5

Activity of the compounds of the present invention against prostate cancer and melanoma

Cell culture and cytotoxicity studies of prostate cancer and melanoma. All cell lines were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA), while cell culture nutrition supplies were purchased from Cellgro Mediatech (Herndon, VA, USA). The present inventors investigated the antiproliferative activity of the proposed anti-tubulin compounds against four human prostate cancer cell lines (LNCaP, DU 145, PC-3 and PPC-1) and three melanoma cell lines (A375, B16-F1 and WM-164). All prostate cancer cell lines were grown in RPMI 1640 supplemented with 10% fetal bovine serum (FBS). Melanoma cells were grown in DMEM supplemented with 5% FBS, 1% antibiotic/antifungal mixture (Sigma-Aldrich, Inc., St. Louis, MO, USA), and bovine insulin (5 μg/ml; Sigma-Aldrich). The cytotoxic potential of anti-tubulin compounds was assessed using a sulforhodamine B (SRB) assay after 96-hour treatment.

Results:

By comparing the proliferation activity data with compound 10, which is an unsubstituted SMART matrix compound, most of the benzo-imidazoles 3-6 had only moderate activity; with the exception of 4, which contains a 5-indolyl as ring substituent A, which had similar activity against the melanoma and prostate cancer cell lines tested (see Table 1). For further modification, 5-indolyl was retained as ring A and imidazole fused to pyridine was used as a replacement for benzoimidazole, giving compound 7. This compound had increased activity compared to 10 and 4. IC ₅₀ values were improved by at least 5 times against A375 melanoma cells and LNCaP androgen-responsive prostate cancer cells. In addition, these new fused ring templates blocked potential phase I metabolic reactions mediated by ketone reduction at 10 (Example 6).

Table 1. Antiproliferative activity of analogues with a fused D-ring matrix

Z IC ₅₀ ± SEM (µM) Melanoma class Prostate cancer cells A375 DU 145 PC-3 LNCaP PPC-1 3 CH Ph BUT. 7.8±0.4 2.4±0.6 2.1±0.3 2.1±0.4 4 CH 5-indolyl 0.025±0.004 0.057±0.005 0.022±0.009 0.028±0.003 0.02±0.01 5 CH 3-indolyl 0.6±0.1 4.2±0.3 0.9±0.2 0.8±0.1 1.3±0.3 6 CH 3-indazolyl 1.1±0.2 4.0±0.1 0.8±0.1 1.6±0.1 1.0±0.1 7 N 5-indolyl 0.005±0.001 BUT. 0.006±0.002 0.005±0.002 0.042±0.003 10 (SMART) - - 0.028±0.005 0.071±0.004 0.021±0.001 0.028±0.004 0.043±0.005

Compound 15 had improved activity compared to trimethoxybenzoyl compound 10 (i.e., 20 nM (15) versus 28 nM (10) against A375 cells; Table 2). This finding supported the assumption that the para position of the benzoyl ring is a suitable site for additional modification. Increasing the chain by two atoms (-OSH ₂ -) by 15 resulted in a compound that retained high activity. However, the idea of alkylation at the para position did not work as expected in inhibiting cancer cell growth. According to the results shown in Table 2, it can be seen that the activity of alkylating agent 12 against melanoma and prostate cancer cells was significantly reduced. Compounds 13 and 14 had similar activity trends as 12. 17, containing a phthalimide protecting group, was active in the micromolar range. The introduction of ethylamine (18) at the para position maintained moderate activity, with an IC ₅₀ value in the hundreds of nanomolar range, but was less active compared to trimethoxybenzoyl compound 10.

Table 2. Antiproliferative activity of analogues containing a benzoyl ring modified at the para position

R IC ₅₀ ± SEM (µM) Melanoma cells Prostate cancer cells B16-F1 A375 DU 145 PC-3 LNCaP PPC-1 12 OCOCH _2Cl 3.2±1.2 10.8±4.4 >10 >10 >10 >10 13 OCOCF ₃ 8.9±2.8 22.2±8.5 >10 >10 >10 >10 14 OCH ₂ Ph 10.6±3.2 >10 >10 >10 >10 >10 15 OCH ₂ OCH ₃ 0.019±0.005 0.020±0.005 0.112±0.017 0.017±0.005 0.031±0.002 0.022±0.005 17 OCH ₂ CH ₂ Phth 1.3±0.3 3.1±0.5 0.6±0.2 >10 1.4±0.8 0.6±0.2 18 OCH ₂ CH ₂ NH ₂ 0.142±0.015 0.527±0.022 0.464±0.03 0.158±0.03 0.117±0.06 0.184±0.02 10 OCH ₃ 0.055±0.005 0.028±0.005 0.071±0.004 0.021±0.001 0.028±0.004 0.043±0.005

* "Phth" refers to phthalimide.

Table 3 shows the results for the compound analogues, where 27 did not demonstrate any activity against all cell lines tested. 33 provided excellent inhibition of prostate cancer and melanoma cell growth in vitro. IC ₅₀ values against prostate cancer cells were improved 2-3 times compared to PAT.

Table 3. Antiproliferative activity of compounds with modified ring A

X n= Ar IC ₅₀ ± SEM (µM) Melanoma cells Prostate cancer cells B16-F1 A375 DU 145 PC-3 LNCaP PPC-1 27 N.H. 1 Ph >30 >30 >30 >30 >30 >30 33 S 1 5-indolyl 0.084±0.016 0.025±
0.006 0.024±
0.005 0.012±
0.002 0.013±
0.004 0.015±
0.001 PAT S 1 Ph 0.065±0.012
0.045±0.008
0.070±0.004
0.057±0.003
0.051±0.001
0.054±0.001
55±0.005 0.028±
0.005 0.071±
0.004 0.021±
0.001 0.028±
0.004 0.043±
0.005

Molecular modeling studies of compound 33 (Figure 7) showed an interaction between this ligand and the α,β-dimer of tubulin via three hydrogen bonds, similar to the interaction of 4 or 7 with tubulin. However, it is clear that the 5-indolyl moiety in compound 33 did not reach the GTP moiety as in the case of 4 or 7, which is possibly caused by the fact that the ketone moiety is not close to the ring system as shown for 4 or 7. Thus , 33 mainly remains in the β-subunit of the tubulin dimer and has a significantly better Glidescore docking index (−8.70).

Example 6

Metabolic stability studies

Microsomal stability study. Metabolic stability studies were carried out by incubating test compounds (0.5 μM) in a reaction mixture with a total volume of 1.2 ml containing 1 mg/ml microsomal protein in reaction buffer [0.2 M phosphate buffer solution (pH 7.4), 1 .3 mM NADP ⁺ , 3.3 mM glucose-6-phosphate and 0.4 U/ml glucose-6-phosphate dehydrogenase] at 37°C in a shaking incubator. Pooled human liver microsomes were used to assess metabolic stability. The NADPH regeneration system (solution A and B) was obtained from Xenotech, LLC (Lenexa, KS, USA). Aliquots (100 μl) of reaction mixtures were removed after 5, 10, 20, 30, 60, and 90 minutes to determine metabolic stability. Acetonitrile (200 μL) containing 200 nM internal standard was added to quench the reaction and precipitate proteins. The samples were then centrifuged at 10,000 rpm for 15 min at RT and the supernatant was directly analyzed by LC-MS/MS (AB Sciex API4500). To identify metabolites, the reaction mixture was incubated for 2 hours together with the test compound at a concentration of 50 μM under the conditions described above. The supernatant was analyzed on a Water Xevo G2-S high-resolution mass spectrometer.

In vitro metabolic stability studies. To determine the possibility of reducing the metabolism of a labile carbonyl linker by incorporating it into a cyclic structure, metabolic stability in liver microsomes was measured using two highly active compounds (4 and 7). The carbonyl linker in 10, which was susceptible to ketone reduction, was replaced with a ring. This modification maintained efficacy while improving metabolic stability by approximately 2-3 times (17 minutes compared to 45 and 51 minutes for human liver microsomes, Table 4) compared to 10.

Table 4. Half-life of the studied compounds from liver microsomes of various types

Connections T _1/2 (min) Human Mouse Rat 4 50.7± 1.2 53.5 ± 2.4 72.3 ± 4.6 7 45.3 ± 2.0 19.7 ± 0.7 30.4 ± 1.9 15 7.8 ± 0.3 4.0 ± 0.3 9.7 ± 0.4 18 110.0± 5.5 123.0± 7.7 225.0 ± 12.6 10 17 <<5 31

In addition, compounds 4 and 7 had increased activity. Metabolic stability of another active analogue 15, containing an increased MOM end group at the para position of the benzoyl ring, was not improved for any of the liver microsomes tested. Another substituent, aminoethyl in 18, also located in the para position relative to O, blocked the tendency to metabolism at the benzoyl ring (T _1/2 ranged from 110 to 225 minutes for the studied types of liver microsomes). The result suggested that the para position of the benzoyl ring may be a site whose modification could lead to improved stability of the compound.

Metabolic pathways of compounds 4, 7, 15 and 18 in vitro. To determine the reason why these new analogues had different metabolic profiles in liver microsomes, additional experiments were performed using a higher concentration (50 μM) of the studied compounds. The present inventors used a high resolution mass spectrometer to identify metabolites where the measured mass error was generally less than 2 ppm. Detailed information regarding the mass spectra and chromatography of each of the metabolites is presented in the Supporting Materials. In case 15 (Figure 8), removal of the MOM group to form M1 (identified by the mass spectrum of 342.08 [M+1]) is the main metabolic pathway, followed by O-demethylation of the 3'- or 5'-methoxy group to form M2 (which was identified by the mass spectrum 328.06 [M+1]). This result was consistent with the short half-life (<10 minutes) of the compound, the MOM group is believed to be unstable following exposure in liver microsomes. M3 (which was identified by a mass spectrum of 372.09 [M+1]) was also an O-demethylation product, however, the exact site of this demethylation was not determined due to the limited information available at this stage. M4 (which was identified by a mass spectrum of 388.12 [M+1]) was a ketone reduction product, then it was hydroxylated to M5 (which was identified by a mass spectrum of 404.12 [M+1]) at a position that is impossible could be determined due to limited information. In case 18 (Figure 9), the main metabolites are M6 (dealkylation) (which was identified by a mass spectrum of 342.08 [M+1]) and M8 (ketone reduction) (which was identified by a mass spectrum of 387.14 [M+1] ]). M7 (deamination) (which was identified by a mass spectrum of 370.11 [M+1]) is a minor product. In case 4 (Figure 10), the main products were O-demethylation (M9) and monohydroxylation (M10). M9 (which was identified by a mass spectrum of 386.15 [M+1]) and M10 (which was identified by a mass spectrum of 416.16 [M+1]) had more than one possible structure, as indicated in the chromatograms. In case 7 (Figure 11), various metabolites were detected, including demethylation products (M12) (which was identified by a mass spectrum of 387.15 [M+1]), mono-hydroxylation (M14) (which was identified by a mass spectrum of 417.16 [M+1]), O-demethylation and subsequent mono-hydroxylation (M11) (which was identified by a mass spectrum of 403.14 [M+1]) and dihydroxylation (M13) (which was identified by a mass spectrum of 433.15 [ M+1]). The indicated products were formed uniformly in similar relative quantities, and all of the indicated metabolites had multiple isomeric forms, as indicated in the chromatograms.

Example 7

Inhibition of tubulin polymerization

Study of tubulin polymerization in vitro. Bovine brain tubulin (0.4 mg, purity >97%) (Cytoskeleton, Denver, CO, USA) was mixed with 10 μM test compound and incubated in 100 μl universal tubulin buffer (80 mM PIPES, 2.0 mM _MgCl2.0 .5 mM EGTA and 1 mM GTP) with pH 6.9. Absorbance at 340 nm was monitored every minute for 20 minutes on a SYNERGY 4 Microplate Reader (Bio-Tek Instruments, Winooski, VT, USA). The spectrophotometer was set at 37°C to polymerize tubulin.

The compounds inhibit tubulin polymerization in vitro. The inhibition of tubulin polymerization was studied using selected highly active compounds 4 and 7, which have improved metabolic properties, and compared with the positive control colchicine and the negative control taxol. DMSO was used as a blank control. Bovine brain tubulin (purity >97%) was incubated with individual compounds (5 or 10 μM) to examine their effect on tubulin polymerization (Figure 12). After 20 min incubation, compound 4 at 5 and 10 µM inhibited tubulin polymerization by 30% and 60%, respectively (Figure 12A) compared to vehicle. Moreover, for compound 7 at a concentration of 5 and 10 μM, 33% and 81% inhibition were observed, respectively (Figure 12B). Both compounds 4 and 7 provided stronger inhibition than colchicine at the two concentrations tested. The data obtained suggested that these compounds have high activity in inhibiting tubulin polymerization, which is in good agreement with their cytotoxicity.

All features described in this application (including the accompanying claims, abstract and any drawings), and/or all steps of any method or process so disclosed can be combined with any of the above aspects in any combination except combinations where at least some of these features and/or steps are mutually exclusive. Although preferred embodiments are shown and described in detail in this application, those skilled in the related fields of technology will understand that various modifications, additions, substitutions, etc., are possible without departing from the spirit of the invention, which, accordingly, are considered to be within the scope of the invention as defined by the following claims.

Claims (15)

1. Compound 15 or 18 or a pharmaceutically acceptable salt thereof, wherein the compound has the formula: 15; or 2. A pharmaceutical composition having anticancer activity and containing an effective amount of a compound according to claim 1 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier. 3. The pharmaceutical composition according to claim 2, further containing at least one pharmaceutically active compound suitable for the treatment of cancer. 4. Use of a compound according to claim 1 for the production of a medicament for the treatment, attenuation, reduction of severity, reduction in the risk of developing or suppressing cancer. 5. The use of claim 4, wherein said cancer is selected from the group consisting of prostate cancer, drug-resistant prostate cancer, breast cancer, drug-resistant breast cancer, ovarian cancer, drug-resistant ovarian cancer, skin cancer, melanoma, drug-resistant melanoma, lung cancer, colon cancer, glioma, leukemia, lymphoma, kidney cancer, central nervous system cancer, uterine cancer, drug-resistant uterine cancer and combinations thereof. 6. Use according to claim 4, characterized in that said cancer is melanoma. 7. Use according to claim 4, characterized in that said cancer is a metastatic melanoma. 8. Use according to claim 4, characterized in that said cancer is prostate cancer. 9. Use according to claim 8, characterized in that said prostate cancer is drug-resistant prostate cancer. 10. Use according to claim 4, characterized in that said medicinal product is intended to be administered in combination with other anticancer therapy. 11. Use of a compound according to claim 1 for the preparation of a medicinal product for the treatment of a drug-resistant tumor or tumors. 12. Use according to claim 11, characterized in that said tumor is selected from the group consisting of prostate cancer, breast cancer, glial tumor, ovarian cancer, skin cancer, melanoma tumor, lung cancer, colon cancer , leukemic tumor, lymphoma tumor, kidney cancer, central nervous system cancer, uterine cancer and combinations thereof. 13. Use according to claim 11, characterized in that said tumor is a melanoma tumor. 14. Use according to claim 11, characterized in that said medicinal product is intended to be administered in combination with other anticancer therapy.