GLUCOSE CONJUGATES OF TRIPTOLIDE, ANALOGS AND USES THEREOF

Abstract

Provided are compounds generated by conjugation of triptolide with glucose to form glucose-triptolide conjugates, provides compounds with effective anti-proliferative activity and improved tolerability as compared to naturally occurring triptolide compounds.

Claims

1. A method of treating cancer in a subject comprising administering to the subject an anti-proliferative effective amount of a glucose-triptolide conjugate compound.

2. The method of claim 1, wherein glucose-triptolide conjugate compound has a structure of Formula I:
T&A-L.sub.1-Sugar wherein the T&A moiety is triptolide or one of its analogs, and can be selected from compounds 1 to 18: ##STR00011## ##STR00012## ##STR00013## wherein L.sub.1 can be selected from XYZ, wherein X and Z can individually and independently be a direct bond, CH.sub.2, C(O), SO, SO.sub.2, OPO, OPO.sub.2, and wherein Y is a substituted or unsubstituted (C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nO(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)O(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nNH(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)NH(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nS(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(CH.sub.2).sub.nS(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nO(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)O(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nNH(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)NH(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nO(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)O(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nNH(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)NH(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkynyl-, wherein each alkyl, alkenyl and alkynyl group may be optionally substituted with alkyl, alkoxy, amino, hydroxyl, oxo, aryl, heteroaiyl, carboxyl, cyano, nitro, or trifluoromethyl; wherein n is an integer independently selected from 0, 1, 2, 3, 4, 5, and 6; wherein the sugar can be selected from compounds 19 to 30, 33 to 48, and 51 to 53: ##STR00014## ##STR00015## ##STR00016## ##STR00017##

3. The method of claim 2, wherein the T&A moiety is compound 1.

4. The method of claim 3, wherein the sugar is compound 19, 20, 37, or 38.

5. The method of claim 4, wherein L.sub.1 is COCH.sub.2CH.sub.2CO or CH.sub.2.

6. The method of claim 5, wherein L.sub.1 is COCH.sub.2CH.sub.2CO.

7. The method of claim 6, wherein the sugar is compound 19.

8. The method of claim 1, wherein the cancer is selected from the group consisting of bladder cancer, breast cancer, ovarian cancer, pancreatic cancer, and gastric cancer, cervical cancer, colon cancer, endometrial cancer, head and neck cancer, lung cancer, melanoma, multiple myeloma, leukemia, non-hodgkin's lymphoma, prostate cancer, rectal cancer, malignant melanomas, alimentary/gastrointestinal tract cancer, liver cancer, skin cancer, lymphoma, kidney cancer, muscle cancer, bone cancer, brain cancer, eye or ocular cancer, rectal cancer, colon cancer, cervical cancer, bladder cancer, oral cancer, benign and malignant tumors, stomach cancer, corpus uteri, testicular cancer, renal cancer, throat cancer, acute lymphocytic leukemia, acute myelogenous leukemia, Ewing's Sarcoma, Kaposi's Sarcoma, basal cell carinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms Tumor, neuroblastoma, mouth/pharynx cancer, esophageal cancer, larynx cancer, neurofibromatosis, tuberous sclerosis, hemangiomas, and lymphangiogenesis.

9. The method of claim 8, wherein the cancer is prostate cancer.

10. The method of claim 8, wherein the cancer is metastatic cancer.

11. The method of claim 1, wherein the compound is administered intravenously.

12. The method of claim 11, wherein the compound is administered at a dosage of about 0.1 mg/kg to 2 mg/kg per dosage.

13. The method of claim 11, wherein the compound is administered once daily.

14. The method of claim 11, wherein the compound is administered for up to about 4 weeks.

15. The method of claim 1, further comprising administering a chemotherapeutic compound.

16. The method of claim 15, wherein the chemotherapeutic compound is administered prior to, simultaneously with, or following administration of the glucose-triptolide conjugate compound.

17. The method of claim 15, wherein the chemotherapeutic compound is selected from the group consisting of taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin and analogs or homologs thereof.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0025] FIGS. 1A-1B. (FIG. 1A) The structure of triptolide and glucose conjugated analogs. (FIG. 1B) The octanol-water partition coefficient logP of different analogs using Interactive logP calculator.

[0026] FIGS. 2A-2D. (FIG. 2A) Effects of G1-5 on HeLa cell proliferation. (FIG. 2B) Effects of G1-5 on ATPase activity of TFIIH in vitro. (FIG. 1C) Effects of G3 and G4 on the stability of RNAPII catalytic subunit. (FIG. 1D) Effects of G1, G2 and G5 on the stability of the RNAPII catalytic subunit.

[0027] FIG. 3. Effects of triptolide and glucose-triptolide conjugate G4 in a metastatic mouse prostate cancer model. The injected prostate cancer cells expressed luciferase, which was detected by imaging of live animals.

DETAILED DESCRIPTION OF THE INVENTION

[0028] The present invention is based on trying to solve the issues of solubility and toxicity associated with triptolide. It was hypothesized that if triptolide could be conjugated to glucose, the two aforementioned problems associated with triptolide could be addressed. The glucose-triptolide conjugates would be preferentially taken up by cancer cells and they should also exhibit much higher water solubility due to the water solubility of glucose moiety.

[0029] As used herein, the term cancer or cancerous growth means the uncontrolled, abnormal growth of cells and includes within its scope all the well-known diseases that are caused by the uncontrolled and abnormal growth of cells. Non-limiting examples of common cancers include bladder cancer, breast cancer, ovarian cancer, pancreatic cancer, and gastric cancer, cervical cancer, colon cancer, endometrial cancer, head and neck cancer, lung cancer, melanoma, multiple myeloma, leukemia (e.g., myeloid, lymphocytic, myelocytic and lymphoblastic leukemias), non-hodgkin's lymphoma, prostate cancer, rectal cancer, and malignant melanomas.

[0030] In addition to invention compounds, one of skill in the art would recognize that chemotherapeutic agents can be used prior to, simultaneously with or following treatment with invention compounds. Illustrative agents include but are not limited to, taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin and analogs or homologs thereof. Therapeutic antibodies or other proteins are also envisioned in combination therapies of the invention.

[0031] The following examples are intended to illustrate but not limit the invention.

EXAMPLE 1

Synthesis of Glucose-Triptolide Conjugates

[0032] This example illustrates the synthesis of glucose-conjugated triptolide compounds. Five glucose conjugates with different types of linkers connecting glucose to triptolide were synthesized, designated G1-G5 (schemes I-III). Based on calculation, the Octanol-water partition coefficient, LogP, values were significantly improved over triptolide itself (FIG. 1).

##STR00004##

##STR00005##

##STR00006##

[0033] The activity of the different glucose-triptolide conjugates was assessed in vitro (FIG. 2). It was found that they differ in their anti-proliferative activity as measured by tritiated thymidine incorporation with G4 and G3 showing moderate activity, while G1, G2 and G5 exhibiting significantly reduced activity. The ability of the glucose conjugates to inhibit the ATPase activity of XPB in the TFIIH complex was also addressed. Interestingly, none of the five glucose conjugates showed appreciable inhibitory activity, suggesting that the cellular activity was a result of breakdown of the conjugates either in cell culture or upon entry into cells. Lastly, the ability of the glucose conjugates to cause degradation of the catalytic subunit of RNAPII was determined, an effect that has been previously observed for triptolide. In parallel with the antiproliferative activity, G4 was most active in inducing RNAPII catalytic subunit degradation with G3 showing weaker activity and the rest of the glucose conjugates showing little activity.

EXAMPLE 2

In Vivo Activity of Glucose-Triptolide Conjugates

[0034] The in vivo antitumor activity of the glucose-triptolide conjugate G4 was determined using an established metastatic prostate cancer model (Reference 1). In this model, the luciferase-expressing prostate cancer line PC3/ML was injected into NOD/SCID/IL2r.sup.null (NSG) mice through the tail vein. The metastasis of the injected prostate cancer cells into liver, kidney, lung and bone can be monitored in live animals by bioluminescent imaging (BLI). It was shown that this is a reliable model with reproducible liver metastasis and all animals succumb by Week 7 after injection of cells (or Week 4 after initiation of treatments). In preliminary experiments, it was found that the tolerable treatment dose of triptolide to be 0.2 mg/kg and that of G4 to be 1 mg/kg. Three weeks after cell injection, each compound was given once daily by IP at those doses for a total of 4 weeks. The surviving animals were continuously monitored upon termination of compound administration. As shown in FIG. 3, mice treated with 11 had lower tumor burden during Weeks 1 and 2 compared with those treated with triptolide. At the end of Week 4, both treatment groups showed undetectable tumor cells while all animals in the untreated groups died. Upon termination of compound administration, tumors immediately returned in animals treated with triptolide. But no tumor cells were detectable in those treated with G4 till the end of the experiment, revealing sustained anticancer activity of G4 in vivo.

EXAMPLE 3

[0035] Other glucose triptolide conjugates with structures similar to Formulas G1-G5 are included in the present invention. Such structures are represented by Formulas G6-G9, wherein R is a direct bond, substituted or unsubstituted (C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nO(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O), substituted or unsubstituted (CH.sub.2).sub.nC(O)(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)O(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nNH(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)NH(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nS(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nO(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)O(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nNH(C.sub.1-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)NH(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH2).sub.nC(O)(CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nO(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)O(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nNH(C.sub.1-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)NH(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkynyl-, wherein each alkyl, alkenyl and alkynyl group may be optionally substituted with alkyl, alkoxy, amino, carboxyl, cyano, nitro, or trifluoromethyl, wherein each n is independently an integer selected from 0, 1, 2, 3, 4, 5, and 6.

##STR00007##

EXAMPLE 4

[0036] Other glucose triptolide conjugates that are included in the present invention are compounds of Formula I:

T&A-L.sub.1-Sugar(I)

[0037] wherein the T&A moiety is triptolide or one of its analogs, and can be selected from compounds 1 to 18:

##STR00008##

[0038] wherein L.sub.1 can be selected from XYZ, wherein X and Z can individually and independently be a direct bond, CH.sub.2, C(O), SO, SO.sub.2, OPO, OPO.sub.2; Y is a direct bond, a substituted or unsubstituted (C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nO(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O), substituted or unsubstituted (CH.sub.2).sub.nC(O)(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)O(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nNH(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)NH(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nS(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(CH.sub.2).sub.nS(C.sub.1-C.sub.6)alkyl-, substituted or unsubstituted (C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nO(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)O(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nNH(C.sub.1-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)NH(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkenyl-, substituted or unsubstituted (C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nO(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)O(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nNH(C.sub.1-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)NH(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkynyl-, substituted or unsubstituted (CH.sub.2).sub.nC(O)(CH.sub.2).sub.nS(C.sub.2-C.sub.6)alkynyl-, wherein each alkyl, alkenyl and alkynyl group may be optionally substituted with alkyl, alkoxy, amino, hydroxyl, oxo, aryl, heteroaryl, carboxyl, cyano, nitro, or trifluoromethyl;

[0039] wherein the sugar can be selected from compounds 19 to 53:

##STR00009## ##STR00010##

[0040] Although the invention has been described with reference to the above example, it will be understood that modifications and variations are encompassed within the spirit and scope of the invention. Accordingly, the invention is limited only by the following claims.

REFERENCES

[0041] The following reference is relied upon and incorporated herein in its entirety.

[0042] 1. Bhatnagar A, Wang Y, Mease R C, Gabrielson M, Sysa P, Minn I, Green G, Simmons B, Gabrielson K, Sarkar S, Fisher P B, Pomper M G. AEG-1 promoter-mediated imaging of prostate cancer. Cancer Res. 2014; 74(20):5772-81.