TREATMENT FOR BREAST CANCER, INCLUDING TRIPLE-NEGATIVE BREAST CANCER

Abstract

The invention relates to the field of medicine and to the chemical and pharmacological industry, and concerns medicaments for the treatment of breast cancer. In particular the invention relates to a use of 3-O-sulfamoyloxy-7β-methyl-D-homo-6-oxa-8α-estra-1,3,5(10)-trien-17a-one as an anti-cancer agent in monotherapy and adjuvant therapy of breast cancer, including the triple negative breast cancer, and a method for the production of the said agent.

Claims

1. A compound, which is 3-O-sulfamoyloxy-7β-methyl-D-homo-6-oxa-8α-estra-1,3,5(10)-trien-17a-one of formula (1): ##STR00005##

2.-3. (canceled)

4. A method of treating cancer, the method comprising administering to a patient in need thereof an effective amount of 3-O-sulfamoyloxy-7β-methyl-D-homo-6-oxa-8α-estra-1,3,5(10)-trien-17a-one of formula (1): ##STR00006##

5. The method of claim 4, wherein the cancer is a breast cancer.

6. The method of claim 5, wherein the patient is a female patient with an early stage of the breast cancer.

7. The method of claim 5, wherein the patient is a female patient with an advanced form of the breast cancer.

8. The method of claim 7, wherein the patient has been treated with tamoxifen.

9. The method of claim 5, wherein the breast cancer is a triple negative form of the breast cancer.

10. The method of claim 6, wherein the female patient is a woman in postmenopause.

11. The method of claim 7, wherein the female patient is a woman in postmenopause.

12. The method of claim 5, wherein 3-O-sulfamoyloxy-7β-methyl-D-homo-6-oxa-8α-estra-1,3,5(10)-trien-17a-one of formula (1) is administered as monotherapy or adjuvant therapy.

13. The method of claim 5, wherein 3-O-sulfamoyloxy-7β-methyl-D-homo-6-oxa-8α-estra-1,3,5(10)-trien-17a-one of formula (1) is administered orally.

14. A method of preparing 3-O-sulfamoyloxy-7β-methyl-D-homo-6-oxa-8α-estra-1,3,5(10)-trien-17a-one of formula (1), the method comprising: i) mixing methyl ether of 7β-methyl-6-oxa-D-homo-8α-estrone of formula (2) ##STR00007##  with hydrobromic acid and acetic acid to obtain 7β-methyl-D-homo-6-oxa-8α-estrone of formula (3): ##STR00008## ii) dissolving the compound of formula (3) in dimethyl formamide followed by the addition of sulfamoyl chloride to obtain a resulting product; iii) extracting the resulting product.

15. The method according to claim 14, wherein acetic acid is a glacial acetic acid, and hydrobromic acid is a 40% HBr.

16. The method according to claim 14, wherein step (i) is carried out under argon atmosphere.

17. The method according to claim 14, wherein the resulting product is extracted with ethyl acetate.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0083] FIG. 1 depicts the staining of breast tumor with ERα antibodies.

[0084] Visualization: horseradish peroxidase+diaminobenzidine, magn. ×100 (A). A specific staining of cell nuclei of mammary duct and nuclei of individual tumor cells (B), magn. ×400.

[0085] FIG. 2 depicts the average volume of tumors at the beginning of the experiment in mice receiving the CAF therapy.

[0086] FIG. 3 depicts the average total volume of tumors at the beginning of the experiment in mice receiving the CAF therapy.

[0087] FIG. 4 depicts the relative change in the average total volume of tumors at the beginning of the experiment in mice receiving the CAF therapy.

[0088] FIG. 5 depicts the relative change in the average total volume of tumors in mice under estimation of antitumor activity of the medicament at the doses of 20 mg/kg and 100 mg/kg.

[0089] FIG. 6 depicts the relative change in the average total volume of tumors in mice when assessing the antitumor activity of the medicament.

[0090] FIG. 7 depicts the relative change of the average total volume of tumors in mice when assessing the anti-tumor activity

EXAMPLES

[0091] The invention is further illustrated by examples of carrying out thereof disclosed herein below.

##STR00004##

Example 1

[0092] The synthesis of 3-O-sulphamoyloxy-7β-methyl-D-homo-6-oxa-8α-estra-1,3,5(10)-trien-17a-one of formula (1).

[0093] To a solution of 300 mg of methyl ether of racemic 7β-methyl-6-oxa-D-homo-8α-estrone of formula (2) in 15 ml of glacial acetic acid 2 ml of 40% HBr was added at the room temperature. The resulting reaction mixture was boiled for 7 h, then poured into water, and extracted with ethyl acetate. The organic layers were washed with water, a saturated NaCl solution, and dried over Na.sub.2SO.sub.4. The solvent was distilled off on a rotatory evaporator. The yield was 174 mg (62%) of racemic 7β-methyl-D-homo-6-oxa-8α-estrone of formula (3) with the melting point of 231-232° C. (the melting point is 230-231° C. according to WO2009059806).

[0094] Elemental analysis, %: C, 76.14; H, 8.17. C.sub.19H.sub.24O.sub.3. Calculated, %: C, 75.97; H, 8.05

[0095] To a solution of 100 mg of the compound of formula (3) in 1 ml of dimethyl formamide 12 mg of sulphamoyl chloride was added. After 2 hours the reaction mixture was poured into 6 ml of a saturated NaCl solution, and the resulting reaction products were extracted with ethyl acetate. The organic phase was stirred with a saturated NaCl solution followed by drying over sodium sulphate. The yield was 93 mg (74%) of the target product of formula (1), m.p. 195-197° C. NMR spectrum .sup.13C (DMSO-d.sup.6), δ, ppm: 214.9, 153.5, 149.8, 125.6, 130.6, 115.0, 111.7, 71.6, 47.4, 44.8, 40.7, 37.9, 34.8, 32.9, 27.4, 26.9, 24.7, 20.0, 19.6. Mass spectrum, m/z (I.sub.rel, %): 379 (100), 331 (18), 316 (6), 305 (11), 278 (62), 270 (72), 251 (48), 225 (32), 211 (43), 199 (55), 185 (22), 171 (30). Elemental analysis, %: C, 60.12; H, 6.48; N, 3.58. C.sub.19H.sub.25NO.sub.5S. Calculated, %: C, 60.14; H, 6.64; N, 3.69.

Example 2

[0096] The cells were seeded on Carrel vials at 50×10.sup.4 cells per flask. To study the proliferative activity of the cells under conditions of sulfatase inhibition, 24 hours after seeding the culture medium was replaced with the medium containing sulfatase inhibitors to a final concentration of 50 μg/ml, and then these tumor cell lines were incubated for various periods of time (24 to 72 hrs). The inhibitor was dissolved in DMSO. The final concentration of DMSO in the culture medium did not exceed 0.5%. To exclude the cytotoxic effect of DMSO, a control sample comprising DMSO without sulfatase inhibitor was prepared. To exclude nonspecific detrimental effects of the compounds, normal human skin fibroblasts were used. Then the cells were detached with the versene-trypsin solution (Biolot), plated onto Carrel vials containing a fresh complete culture medium. The cells were counted when untreated control cells reached the maximum cell density per unit of surface area of a culture flask (monolayer), and their number was set as 100%. The proliferative activity of all of the studied cell cultures exposed to sulfatase inhibitors was determined in triplicate.

[0097] The study of the obtained sulfamate effects on the proliferation of the passaged human MCF-7 cell culture (breast adenocarcinoma) has demonstrated that the compound of formula (1) at a concentration of 20 μg/ml completely blocks the proliferation of tumor cells, but it does not influence the growth of human skin fibroblasts having no estrogen receptors. The proliferation of tumor cells is inhibited to the same extent as under the action of Tamoxifen, which has been used in clinical practice for more than 30 years. This is very important because sulfamates and Tamoxifen have different mechanisms of action The experimental data were analyzed with statistical software package “STATISTICA 5.0”.

TABLE-US-00010 TABLE 10 Effect of the agent on the biochemical parameters of ovariectomized rats Rat group Change in body Triglyceride (number of weight during Ash femur Cholesterol level level in blood animals in the the experiment, Uterus weight, weight/“Wet” in blood serum, serum, mg/ group) g mg/100 g bw femur weight mg/100 ml 100 ml Sham operated 29 ± 3* 200 ± 12** 0.435 ± 0.005* 50.4 ± 2.9* 85.4 ± 5.9* (10) Ovariectomised 54 ± 4  32 ± 3   0.400 ± 0.003  69.3 ± 3.2  69.2 ± 5.4  (15) Ovariectomised,  3 ± 4* 164 ± 11** 0.436 ± 0.004* 45.5 ± 2.1* 129.8 ± 14.5* treated with EE (0.1 mg/kg bw) (15) Ovariectomised, 48 ± 4* 33 ± 3   0.390 ± 0.004  52.2 ± 2.6* 59 59.6 ± 4.8     treated with the medicament (5 mg/kg bw) (15) Note. EE is 17α-ethinyl estradiol. *P .sub.ovariectomized < 0.05; **P .sub.ovariectomized < 0.01.

[0098] The findings demonstrate that the claimed agent has no uterotropic activity, displays a hypocholesteremic action without raising triglyceride levels in the blood serum. The latter is important because high triglyceride levels in serum are associated with a risk of cardiovascular diseases.

[0099] In experiments on FVB mice, transgenic for HER-2/neu (ER−/PR−/HER2+), with breast tumors, the agent inhibits tumor growth to the same extent as clinically approved Tamoxifen, which suggests their use in a combination for the treatment of breast cancer (this might result in lowering of the side effects).

[0100] On the MDA-MB-231 line of cells of triple-negative breast cancer the compound inhibits the cell growth, IC.sub.50=4.8 μM, which is similar to Etoposide, a clinically approved chemotherapeutic medicament.

[0101] It was demonstrated that the chemotherapy of patients having metastases spread into axillary lymph nodes with the present medicament reliably prolongs recurrence-free survival rate by 11% as compared to the untreated group and increase the overall survival rate as compared to the Tamoxifen treated group.

[0102] On animal models with an passaged human tumor of triple-negative breast cancer, the medicament showed a better action as compared to clinically approved Tamoxifen and an aromatase inhibitor such as Letrozole.

INDUSTRIAL APPLICABILITY

[0103] As a result of the conducted experiments it was found that the present compound demonstrates activity against triple-negative breast cancer, is nontoxic, has a hypocholesteremic action, has no effect on triglyceride levels in blood serum, does not adversely affect the endometrium, has no uterotropic action.

[0104] In view of its properties mentioned above, the present compound can be used for: [0105] the monotherapy, and adjuvant therapy for early hormone-positive breast cancer in postmenopausal women, [0106] the treatment of triple-negative breast cancer, [0107] the monotherapy and adjuvant therapy of breast cancer in postmenopausal women having been treated with Tamoxifen for 2- or 3-years. [0108] the treatment for advanced breast cancer at late stages.