Use of KIF18a inhibitor in treatment of breast cancer

Abstract

A use of a KIF18A inhibitor in the preparation of a drug for treating breast cancer. Pharmacological experiments demonstrate that compound 1 can inhibit the proliferation of breast cancer cells and inhibit the growth of heterogeneous tumors of triple-negative breast cancer. The results indicate that the compound 1 has a significant therapeutic effect on breast cancers (including triple-negative breast cancer).

Description

Use of KIF18A inhibitor in treatment of breast cancer

[0001] This application claims the priority of the prior application with the following details:

[0002] The priority of the prior application with the following details: the prior application with the patent application number 202410864749.4, entitled "Use of KIF18A inhibitor in treatment of breast cancer", filed with the China National Intellectual Property Office on June 28, 2024, is claimed in this application, and the prior application is incorporated herein by reference in its entirety. TECHNICAL FIELD

[0003] The present application belongs to the field of medicine, and specifically relates to the use of a KIF18A inhibitor in the preparation of a medicament for treating breast cancer. BACKGROUND

[0004] Cancer is one of the most serious diseases affecting human health, and the mortality and morbidity rates are often among the top of various diseases. Although the life quality of some patients has been greatly improved with the continuous development and progress of medical technology and drug research and development, there are still more unmet clinical needs in the search for effective drugs for treating or curing different cancers, and more new targets will provide new possibilities for future cancer drug research and development.

[0005] Due to the damage or deletion of one or more genes regulating the cell cycle, unregulated cell proliferation occurs in cancer cells. Various kinases and kinesins have been identified to play a key role in the regulation and progression of cell cycle and mitosis of normal dividing cells and cancer cells.

[0006] Kinesin molecules are motor proteins that use intracellular microtubules as tracks, also known as molecular motors, which can convert ATP energy into mechanical energy, and are closely related to mitosis and meiosis of eukaryotic cells, growth and development of tissues and organs, development and signal transduction of neurons, etc. Kinesin members share a relatively conserved motor domain. According to the position of the motor domain in the molecule, the kinesin family is roughly divided into three types: N-type kinesin, i.e., the amino (-NH2) terminal region of the protein polypeptide chain has a motor domain; M-type kinesin, i.e., the middle region has a motor domain; C-type kinesin, i.e., the carboxyl (-COOH) terminal region has a motor domain.

[0007] KIF18A is a member of the N-type Kinesin-8 kinesin family. KIF18A protein is highly expressed in various tumors. Therefore, the development of KIF18A protein inhibitors may be a new breakthrough in cancer treatment drugs.

[0008] Breast cancer is a common malignant tumor in women. There is a definite clinical need to develop drugs for effectively treating breast cancer. SUMMARY

[0009] In order to improve the deficiencies of the prior art, the purpose of the present application is to provide a drug having a definite therapeutic effect on breast cancer.

[0010] The purpose of the present application is achieved by the following technical solutions.

[0011] The present application provides the use of Compound 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating breast cancer, wherein the structure of Compound 1 is as follows:

[0012] According to an embodiment of the present application, the pharmaceutically acceptable salt is selected from one or more of a sodium salt, a potassium salt, a hydrochloride salt, a p-toluenesulfonate salt, a maleate salt, and a methanesulfonate salt of Compound 1, and is preferably a p-toluenesulfonate salt.

[0013] According to an embodiment of the present application, the medicament comprises a pharmaceutically acceptable carrier or excipient, and is suitable for preparation into a form of intravenous infusion, intravenous drip, subcutaneous administration, intradermal administration, intramuscular injection, oral spray, oral administration, in situ administration to a tumor, etc.

[0014] According to an embodiment of the present application, the medicament is prepared into an oral preparation, such as a tablet, a capsule, a pill, a granule, a solution, a suspension, a syrup, an injection (including an injection solution, a sterile powder for injection, or a concentrated solution for injection), a suppository, an inhalant, or a spray.

[0015] According to an embodiment of the present application, the medicament is administered orally.

[0016] According to an embodiment of the present application, the medicament is administered once a day.

[0017] The present application also provides a method of preventing and / or treating breast cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of Compound 1 or a pharmaceutically acceptable salt thereof, or a composition containing Compound 1 or a pharmaceutically acceptable salt thereof.

[0018] According to an embodiment of the present application, the pharmaceutically acceptable salt is selected from one or more of a sodium salt, a potassium salt, a hydrochloride salt, a p-toluenesulfonate salt, a maleate salt, and a methanesulfonate salt of Compound 1, and is preferably a p-toluenesulfonate salt.

[0019] According to an embodiment of the present application, the breast cancer is ductal carcinoma of the breast, invasive ductal carcinoma, lobular carcinoma, or triple-negative breast cancer, and is preferably triple-negative breast cancer.

[0020] According to an embodiment of the present application, the breast cancer is TP53 mutant breast cancer.

[0021] According to an embodiment of the present application, the breast cancer is TP53-mutated triple negative breast cancer.

[0022] According to an embodiment of the present application, the breast cancer is recurrent or metastatic advanced breast cancer.

[0023] According to an embodiment of the present application, the breast cancer is TP53-mutated recurrent or metastatic advanced breast cancer.

[0024] According to an embodiment of the present application, the compound 1 is administered alone or in combination with other anti-tumor drugs.

[0025] According to an embodiment of the present application, the compound 1 or its pharmaceutically acceptable salt or the composition is administered once a day.

[0026] According to an embodiment of the present application, the compound 1 or its pharmaceutically acceptable salt or the composition is administered orally.

[0027] According to an embodiment of the present application, the combination administration can be simultaneous or sequential administration.

[0028] According to an embodiment of the present application, the daily administration dose of the compound 1 is 0.1-1000 mg, which can be 0.1 mg, 0.3 mg, 0.5 mg, 0.7 mg, 0.9 mg, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 825 mg, 850 mg, 875 mg, 900 mg, 925 mg, 950 mg, 975 mg, 1000 mg.

[0029] According to an embodiment of the present application, the subject is a mammal, preferably a human.

[0030] The present application also provides a medicament for preventing and / or treating breast cancer disease in a subject, the medicament being Compound 1 or a pharmaceutically acceptable salt thereof, or the medicament being a composition containing a therapeutically effective amount of Compound 1 or a pharmaceutically acceptable salt thereof, the structure of Compound 1 being shown as follows:

[0031] According to an embodiment of the present application, the pharmaceutically acceptable salt is selected from one or more than two of sodium salt, potassium salt, hydrochloride salt, p-toluenesulfonic acid salt, maleic acid salt and methanesulfonic acid salt of Compound 1, preferably the p-toluenesulfonic acid salt.

[0032] Advantages of the present application:

[0033] The pharmacological experiments of the present application demonstrate that Compound 1 can inhibit the proliferation of breast cancer cells, inhibit the growth of triple-negative breast cancer heterograft, and show that Compound 1 has obvious therapeutic effect on breast cancer (including triple-negative breast cancer). BRIEF DESCRIPTION OF DRAWINGS

[0034] Figure 1: Line graph of the influence of tumor volume in MX-1 xenograft model mice.

[0035] Figure 2: Diagram of the tumor weight of MX-1 xenograft model mice.

[0036] Figure 3: Line graph of the influence of body weight of MX-1 xenograft model mice.

[0037] Definitions and explanations of terms

[0038] The term "subject" herein refers to an animal, such as a mammal. Mammals include, for example, mice, rats, dogs, cats, pigs, sheep, horses, cows, and humans.

[0039] The term "therapeutically effective amount" or "effective amount" herein refers to the amount of a compound disclosed and / or described herein that, when administered to a patient in need of such treatment, is sufficient to effect treatment as defined herein. The therapeutically effective amount of a compound can be an amount sufficient to treat a disease responsive to modulation (e.g., inhibition) of KIF18A or an amount sufficient to treat breast cancer. The therapeutically effective amount will vary depending on, for example, the subject and disease condition being treated, the subject's body weight and age, the severity of the disease condition, the particular compound, the dosing regimen to be followed, the timing of administration, the mode of administration, all of which can be readily determined by one of ordinary skill in the art.

[0040] The term "treatment" herein includes one or more of: inhibiting the disease or condition; slowing or arresting the development of a clinical symptom of the disease or condition; and / or relieving the disease or condition (i.e., causing regression or remission of a clinical symptom), and complete or partial reduction of a clinical symptom of the disease or condition.

[0041] The term "therapeutic effect" herein means an effect caused by treatment, which is manifested at the cellular level as the inhibition rate of cell growth or the mortality rate of cells, at the animal level as the change, usually the alleviation or improvement of the symptoms of a disease or a disease condition, or the cure of a disease or a disease condition.

[0042] The term "pharmaceutically acceptable salt" herein means that the compound can exist in the form of various pharmaceutically acceptable salts. If the compound has a basic center, it can form an acid addition salt; if the compound has an acidic center, it can form a base addition salt; if the compound contains both an acidic center and a basic center, it can also form an internal salt. DETAILED DESCRIPTION

[0043] The present application will be further described in conjunction with specific examples. It should be understood that the following examples are merely illustrative and explanatory of the present application and should not be construed as limiting the scope of the present application. Any technology realized based on the above description of the present application is encompassed within the scope intended to be protected by the present application.

[0044] The specific preparation method of Compound 1 is described in the patent document of WO2024051755A1.

[0045] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0046] Example 1

[0047] 1.1 Purpose of the experiment

[0048] CTG method was used to evaluate the effect of Compound 1 on the cell proliferation of breast cancer cell lines.

[0049] 1.2 Abbreviation table

[0050] 1.3 Materials

[0051] 1.3.1 Cell lines

[0052] 1.3.2 Reagents and consumables

[0053] (1) Cell culture routine culture medium and consumables;

[0054] (2) Fetal Bovine Serum FBS (ExCell Bio., Cat# FND500);

[0055] (3) CellTiter-Glo Luminescent Cell Viability Assay (Promega, Cat# G7573);

[0056] (4) 96-well black-walled clear-bottom cell culture plates (Corning, Cat# 3904).

[0057] 1.3.3 Instruments

[0058] EnVision Multi-label Microplate Reader, PerkinElmer, 2104-0010A;

[0059] Ultra Micro Digital Pipettor, Tecan, D300e;

[0060] Cell Counter, Inno-Alliance Biotech, Countstar;

[0061] CO2 Incubator, Thermo Scientific, Model 3100 Series;

[0062] Biosafety Cabinet, Thermo Scientific, Model 1300 Series A2;

[0063] Inverted Microscope, Olympus, CKX41SF;

[0064] Refrigerator, SIEMENS, KK25E76TI.

[0065] 1.4 Test Samples

[0066] AMG650, Batch No.: EB2302070-048P1; Storage Condition: 2-8℃; Production and Sample Sending Unit: Changchun Jin Sai Pharmaceutical Co., Ltd.

[0067] Compound 1, Batch No.: EB2214099-062P1; Storage Condition: 2-8℃; Production and Sample Sending Unit: Changchun Jin Sai Pharmaceutical Co., Ltd.

[0068] Cisplatin; Produced by Qilu Pharmaceutical, Storage Condition: Room Temperature.

[0069] Preparation Method: A suitable amount of test sample was accurately weighed into a suitable container, DMSO was added, and ultrasonic vortex was performed to obtain a clear solution for administration. The prepared solution was stored at -20℃. Before administration, it was placed at room temperature.

[0070] 1.5 Experimental Methods

[0071] 1.5.1 Cell Culture

[0072] Cells were thawed and cultured in their respective media.

[0073] Cell plating (Day 1): Cells in log phase of growth were harvested and counted using a cell counter. Cell viability was determined by trypan blue exclusion to ensure that the viability of each cell line was > 85%. The cell concentration was adjusted by dilution with media and 135 μL of the cell suspension was added to a 96-well cell plate to achieve the specified cell density (including the cell control TO on the day of drug treatment). The cells in the 96-well plate were incubated overnight at 37°C, 5% CO2, and 95% humidity.

[0074] TO cell viability readout (Day 2): The CellTiter-Glo reagent and cell culture plate were equilibrated at room temperature for 30 minutes.

[0075] Fifteen μL of media was added to each well, followed by 75 μL of CellTiter-Glo reagent. The cells were allowed to lyse thoroughly by shaking on an orbital shaker for 2 minutes. The cell culture was equilibrated at room temperature for 10 minutes. The chemiluminescent value was read on an EnVision.

[0076] Drug treatment (Day 2): Test compounds were dissolved in DMSO to make stock solutions and were serially diluted to make 10-fold working concentration solutions; similarly, 10-fold solutions of positive drugs were prepared. Fifteen μL of drug solution was added to each well of the 96-well plate that had been seeded with cells, and three replicates were set up for each cell concentration. The highest concentration of test compound was 10 μM, with nine concentrations, 3-fold dilutions. The cells in the 96-well plate with added drug were incubated for an additional 168 hours at 37°C, 5% CO2, and 95% humidity.

[0077] Cell viability readout (Day 9): The CellTiter-Glo reagent and drug-treated cell culture plate were equilibrated at room temperature for 30 minutes. Seventy-five μL of CellTiter-Glo reagent was added to each well. The cells were allowed to lyse thoroughly by shaking on an orbital shaker for 2 minutes. The cell culture was equilibrated at room temperature for 10 minutes. The chemiluminescent value was read on an EnVision.

[0078] 1.5.2 Data Analysis

[0079] Data were analyzed using GraphPad Prism 5.0 or Dataportal software, and a non-linear S-curve regression was used to fit the data to obtain a dose-effect curve, from which the IC 50 values were calculated.

[0080] Cell survival rate (%) = (Lum test drug - Lum culture solution control) / (Lum cell control - Lum culture solution control) x 100%.

[0081] Lum cell control - Lum culture solution control is set to 100%, and the Lum Medium control value is set to 0%.

[0082] 1.6 Experimental results

[0083] As shown in Table 1, compared with AMG650 and cisplatin, compound 1 has stronger inhibitory effect on the proliferation of TP53 mutant breast cancer cells, and compound 1 has significant inhibitory effect on various breast cancer cells.

[0084] Table 1 Inhibitory effect of compound 1 on the proliferation of breast cancer cells

[0085] Example 2

[0086] Experimental name Pharmacodynamic effect test of compound 1 on human triple-negative breast cancer MX-1 cell xenograft tumor mice

[0087] 2.1 Purpose of the experiment

[0088] To observe the therapeutic effect of compound 1 on human triple-negative breast cancer MX-1 cell xenograft tumor mice and evaluate its effect.

[0089] 2.2 Experimental materials

[0090] 2.2.1 Experimental reagents

[0091] 2.2.2 Experimental instruments

[0092] 2.3 Test drug

[0093] Compound 1, batch number: CE140A.2TsOH-KL2401001; storage condition: 15-25°C; production and sample sending unit: Changchun Jin Sai Pharmaceutical Co., Ltd.

[0094] Solvent components: 20% PEG400 + 10% VE-TPGS + 70% aqueous solution containing 10% hydroxypropyl-β-cyclodextrin.

[0095] The preparation method is as follows: accurately weigh an appropriate amount of test product into a suitable container, and add the above solvent components step by step, and ultrasonically vortex to obtain a clear solution after each step, then add the next step, and the final solution after preparation is a clear and transparent solution for administration.

[0096] Storage condition of the administration solution: The solution was stored at -20℃ after aliquotting. The solution was placed at room temperature before administration.

[0097] 2.4 Cell lines

[0098] Human triple negative breast cancer cell MX-1: purchased from ATCC by Beijing Aisipu Biotechnology Co., Ltd.

[0099] 2.5 Experimental animals

[0100] Species and strain: Balb / c Nude mice.

[0101] Gender and age: female, 6-9 weeks old.

[0102] Source of animals: Beijing Vantoll Life Experimental Animal Technology Co., Ltd. The animal qualification certificate is: 110011230106884731.

[0103] Housing conditions: housed in polypropylene mouse boxes, with a maximum of 5 animals per box. Temperature: set temperature range 20-26℃, humidity: set humidity range 40%-70%, air changes: not less than 15 times fresh air / hour, light: 12 hours light and 12 hours dark alternately.

[0104] Establishment of animal models: human triple negative breast cancer cell MX-1 was cultured in vitro, and the in vitro culture conditions were as follows: RPMI 1640 medium containing 10% fetal bovine serum (FBS) and 1% antibiotic-antimycotic (antibiotic-antimycotic), temperature 37℃, cultured in a 5% CO2 cell incubator. Routine medium replacement was performed twice a week, and cells in the logarithmic growth phase were collected, counted and used for tumor inoculation. The MX-1 cells were collected, resuspended in an equal proportion of mixed medium and matrigel mixture to a concentration of 1*10^ 8 / mL, and transplanted in the right front subcutaneous position of the mouse shoulder and back at a volume of 100 μL at a dose of 1*10^ 7 / each. After inoculation, the mice were randomly divided into groups when the average tumor volume reached 130.98±2.57mm 3 .

[0105] The experimental groups are as follows:

[0106] According to the relevant regulations of animal welfare, if an individual experimental animal meets any of the following conditions during the experiment, the animal will be removed from the experimental group and euthanized. (1) The animal's body weight decreases by more than 20% compared to Day 0 (BWL≥20%) and is scheduled for euthanasia; (2) the animal has severe adverse reactions such as blindness, paralysis, etc.; (3) the tumor volume is greater than 2000mm 3(4) Open ulceration on the tumor surface.

[0107] Blood and tumor tissue were collected after the last administration for weighing, photographing and quick-freezing preservation.

[0108] 2.6 Experimental observation index and detection method

[0109] During the experiment, the animal body weight and tumor volume were measured twice a week, and the animal clinical symptoms were observed and recorded daily.

[0110] The tumor volume (TV) calculation formula was: 1 / 2×a×b 2 , wherein a and b were the length and width of tumor measurement, respectively.

[0111] The tumor inhibition rate (%TGI TV ) calculation formula was: 1-(TV Tn -TV T0 ) / (TV Cn -TV C0 )*100%, TV C was the average tumor volume of the negative control group, and TV T was the average tumor volume of the treatment group.

[0112] The animal body weight change (%BWC) calculation formula was: (BW t -BW0) / BW0×100%, wherein BW t was the animal body weight at each measurement, and BW0 was the animal body weight at grouping.

[0113] According to the Chinese NMPA “Technical Guidelines for Nonclinical Studies of Cytotoxic Antitumor Drugs” (November 2006), %T / C≤40% and P<0.05 by statistical analysis were effective. If the number of drug-related animal deaths exceeded 20%, the drug dose was considered to have serious toxicity.

[0114] 2.7 Statistical analysis

[0115] One-way ANOVA was used for analysis between groups. No significant difference was considered when p>0.05, significant difference was considered when p<0.05, obvious significant difference was considered when p<0.01, and extremely significant difference was considered when p<0.001.

[0116] 2.8 Experimental results

[0117] As shown in Figure 1, the TGI of compound 1 at 75 mg / kg, 100 mg / kg and 150 mg / kg was 60.28%, 71.80% and 66.86%, respectively.

[0118] As shown in Figure 2, the tumor weight of the model group mice was 942 ± 91 mg, and the tumor weights of the Compound 1 75 mg / kg, 100 mg / kg and 150 mg / kg treatment groups were 390 ± 59 mg, 360 ± 73 mg and 382 ± 55 mg, respectively.

[0119] As shown in Figure 3, all dose groups had no significant effect on body weight changes relative to the vehicle group animals, and no obvious toxicity was observed.

[0120] In summary, in the MX-1 triple-negative breast cancer xenograft mouse model, Compound 1 (75 mg / kg, 100 mg / kg and 150 mg / kg) significantly inhibited tumor growth, showing a significant therapeutic effect on triple-negative breast cancer.

[0121] The above describes embodiments of the present application. However, the present application is not limited to the above-described embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the scope of protection of the present application.

Claims

1. Use of Compound 1 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of breast cancer, wherein, The structure of the compound 1 is shown below: Preferably, the pharmaceutically acceptable salt is selected from one or more of sodium salt, potassium salt, hydrochloride, p-toluenesulfonate, maleate and mesylate of Compound 1, preferably p-toluenesulfonate.

2. Use according to claim 1, wherein, The breast cancer is ductal carcinoma of breast, invasive ductal carcinoma, lobular carcinoma or triple negative breast cancer, preferably triple negative breast cancer.

3. Use according to claim 1, wherein, The breast cancer is TP53 mutant breast cancer, preferably TP53 mutant triple negative breast cancer.

4. Use according to any one of claims 1 to 3, wherein, The medicament comprises a pharmaceutically acceptable carrier or excipient, and is suitable for preparation into intravenous infusion, intravenous drip, subcutaneous administration, intradermal administration, intramuscular injection, oral spray, oral administration, in situ administration to tumor, etc. Preferably, the medicament is administered orally, more preferably, the medicament is administered once a day.

5. Use according to any one of claims 1 to 4, wherein, The medicament is prepared into oral preparations, such as tablets, capsules, pills, granules, solutions, suspensions, syrups, injections, suppositories, inhalants or sprays.

6. A method of preventing and / or treating breast cancer disease in a subject, wherein, The methods include administering to the subject a therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a composition containing Compound 1, or a pharmaceutically acceptable salt thereof, the structure of which is shown below: Preferably, the pharmaceutically acceptable salt is selected from one or more of sodium salt, potassium salt, hydrochloride, p-toluenesulfonate, maleate and mesylate of Compound 1, preferably p-toluenesulfonate; preferably, Compound 1 or its pharmaceutically acceptable salt or the composition is administered once a day, more preferably, Compound 1 or its pharmaceutically acceptable salt or the composition is administered orally.

7. The method of claim 6, wherein, The breast cancer is ductal carcinoma of breast, invasive ductal carcinoma, lobular carcinoma or triple negative breast cancer, preferably triple negative breast cancer, more preferably the breast cancer is TP53 mutant breast cancer, further preferably the breast cancer is TP53 mutant triple negative breast cancer.

8. The method of claim 6 or 7, wherein, Compound 1 or its pharmaceutically acceptable salt, or a composition containing Compound 1 or its pharmaceutically acceptable salt, is administered alone or in combination with other anti-tumor drugs, and the combination administration can be simultaneous or sequential administration.

9. The method according to any one of claims 6-8, wherein, The daily administration dose of the compound 1 is 0.1-1000 mg, which can be 0.1 mg, 0.3 mg, 0.5 mg, 0.7 mg, 0.9 mg, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 825 mg, 850 mg, 875 mg, 900 mg, 925 mg, 950 mg, 975 mg, 1000 mg.

10. The method according to any one of claims 6-9, wherein, The subject is a mammal, preferably a human.

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