Anti-tumor small molecule compound and preparation method and application thereof

Abstract

The invention relates to an anti-tumor small molecule compound as well as a preparation method and application thereof. The invention relates to an anti-tumor compound or a pharmaceutically acceptable salt and a stereoisomer thereof, and a pharmaceutical composition containing the anti-tumor compound. According to the present invention, the compound is phenethyl 5-hydroxy-6-(((3R, 4S, 5S, 6R)-3, 4, 5-trihydroxy-6-(hydroxymethyl) tetrahydro-2H-pyran-2-yl) oxy)-2-naphthoic acid ester by using the IUPAC standard naming method, has been proved to have a good anti-tumor effect, and can effectively inhibit the proliferation of a variety of tumor cells. The invention also relates to a preparation method of the compound and application of the compound in preparation of medicines for preventing and / or treating tumors.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to a compound capable of inducing the death of various tumor cells, and a preparation method and use thereof. Background Art

[0002] Tumor is an aggregate of abnormal cells. Compared with the corresponding normal cells, tumor cells can achieve self-sufficiency of growth signals, are insensitive to growth inhibitory signals in the body, and show unrestricted growth ability and replication potential. Tumor cells generally have the ability to promote angiogenesis in the surrounding area, evade programmed cell death, and the abilities of tissue invasion and metastasis also exacerbate their malignant growth degree. At present, various tumors including liver cancer, lung cancer, melanoma, and colon cancer have been widely characterized. Certain clinical effects can be achieved through existing tumor treatment methods, and the survival time of patients can be extended. However, more than 9.7 million people died from tumors globally in 2022, and the incidence rate is increasing year by year. Therefore, tumors remain a global problem seriously threatening human life and health.

[0003] Primary liver cancer (PLC) is a typical malignant tumor, characterized by difficult early diagnosis, rapid progression, poor prognosis, and high mortality. Hepatocellular carcinoma (HCC) is a major subtype of primary liver cancer, accounting for about 85% of primary liver cancer. According to the results of epidemiological investigations, hepatitis B virus (HBV) is a key factor leading to hepatocellular carcinoma. In addition, hepatitis C virus (HCV), aflatoxin, alcohol, and nitrosamine compounds are all factors inducing liver cancer. At present, the main treatment methods for hepatocellular carcinoma include liver transplantation, hepatectomy, radiofrequency ablation, radiotherapy, and chemotherapy. When liver cancer develops to the advanced stage, targeted drugs represented by sorafenib and lenvatinib have the best efficacy. In recent years, with the rise of immune checkpoint inhibitors and monoclonal antibodies, such as PD-L1 monoclonal antibody, has also made progress in clinical practice. However, the first-line drugs for liver cancer at the present stage still have limitations such as limited selection types, strong toxic and side effects, poor prognosis, and easy generation of drug resistance. Therefore, there is an urgent need for effective new anti-tumor drugs or new drug combinations clinically.

[0004] Lung cancer generally refers to primary bronchogenic carcinoma, which is the most common malignant tumor of the lungs and the cancer with the highest incidence in China. It includes small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). Among them, NSCLC accounts for 80% of the total cases, featuring high mortality and strong heterogeneity. The incidence of lung cancer is closely related to factors such as smoking, air pollution, occupational exposure, genetic factors, and other chronic pulmonary diseases. According to the size of the primary tumor, whether lymph node metastasis and other organ invasions occur, lung cancer can be divided into stages I to IV according to the TNM staging system. For patients with early and middle-stage lung cancer, surgical treatment is the preferred option, and other treatment methods include chemoembolization and radiotherapy. According to whether key tumor target gene mutations such as EGFR occur in lung cancer patients, the drug treatment of lung cancer mainly focuses on tyrosine kinase inhibitors such as gefitinib, erlotinib, and osimertinib, effectively prolonging the overall survival of patients. However, generally speaking, NSCLC is less sensitive to chemotherapy and radiotherapy, and the development of new anti-cancer drugs remains the top priority in the research of lung cancer treatment.

[0005] Melanoma is a malignant tumor originating from melanocytes and is the most dangerous skin cancer. It can develop from existing moles or be a new lesion. Its early symptoms are not obvious and are not easily noticed by patients. However, it has a high degree of malignancy and is prone to lymph node and distant metastasis. After metastasis occurs, the survival period of melanoma patients will drop significantly, so it is characterized by a high metastasis rate and high mortality. Epidemiological studies show that the affected population is mainly middle-aged and elderly men, and the incidence and mortality rates in men are higher than those in women. The incidence of melanoma is mainly related to DNA damage caused by sunlight or ultraviolet rays, genetic pathogenic genes, and local stimuli such as repeated friction and trauma. For early-stage patients, Mohs surgery and other surgeries can achieve a complete cure. Drug treatment methods include chemotherapy treatments such as dacarbazine and paclitaxel, as well as targeted drug treatments for patients with cancer gene mutations such as BRAF and MEK, including dabrafenib and trametinib. The treatment targeting the above single targets is likely to promote the development of drug resistance in melanoma. Developing new types of anti-cancer compounds is expected to improve the drug resistance of melanoma and enhance the treatment effect.

[0006] Colorectal Cancer (CRC), also known as colon cancer or rectal cancer, refers to tumors that develop from the colon or rectum. It is the second most common cancer in China, with over 500,000 patients, and the incidence is on the rise. Colon cancer has the characteristics of inconspicuous early symptoms and slow disease progression. Most patients are diagnosed at the middle or late stage, so it also has the characteristics of limited treatment methods and poor prognosis. Its causes are related to age, genetic factors, and chronic inflammations such as ulcerative colitis, and are closely related to eating habits such as alcoholism, high-fat, high-red meat, and pickled food intake. The main diagnostic method for colon cancer is colonoscopy, which detects the degree of disease progression and whether it has spread through sampling and medical imaging. According to the diagnosis results, colon cancer can be treated by endoscopic surgical resection, chemotherapy such as capecitabine and fluorouracil, and radiotherapy. Based on its epigenetic changes, targeted therapies such as cetuximab and bevacizumab can be used to effectively extend the survival period of patients. However, the mechanism of colon cancer chemotherapy resistance is complex and is prone to treatment failure through mutations such as KRAS and activation of signaling bypasses.

[0007] Therefore, the development of novel spectral anti-tumor compounds is of great significance for the research of new drugs or drug combinations for the treatment of cancers such as hepatocellular carcinoma. Summary of the Invention

[0008] Object of the Invention: The object of the present invention is to provide a compound with the structure of phenethyl 5-hydroxy-6-(((3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-2-naphthoate (hereinafter simply referred to as WX003) or its pharmaceutically acceptable salts and stereoisomers:

[0009]

[0010] In some preferred embodiments, the pharmaceutically acceptable salts include acid addition salts formed by this compound and the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid; it also includes acid salts formed by this compound and inorganic bases.

[0011] In some more preferred embodiments, the pharmaceutically acceptable salts include alkaline metal cation salts, alkaline earth metal cation salts, and ammonium cation salts.

[0012] The compounds involved in the present invention can also exist in the form of their salts, which are converted into the compounds mentioned in this patent in vivo. For example, within the scope of the present invention, according to the processes known in the art, the compounds of the present invention are converted into the form of pharmaceutically acceptable salts and used in the form of salts.

[0013] All tautomeric forms of the compounds of the present invention are included within the scope of the present invention. The compounds of the present invention may exist in specific geometric or stereoisomeric forms. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups, and all such isomers and their mixtures are included within the scope of the present invention.

[0014] The WX003 and its derivatives of the present invention can be prepared by the above-mentioned or similar preparation methods, and the corresponding starting materials can be selected according to different substituents. Those skilled in the art should recognize that the above routes are helpful for understanding the present invention, but do not limit the content of the present invention. Unless otherwise specified, the variables are defined as mentioned in general formula I.

[0015] Another object of the present invention is to provide a pharmaceutical composition, which comprises the compound of the present invention or its pharmaceutically acceptable salt, stereoisomer, and a pharmaceutically acceptable carrier or excipient.

[0016] The pharmaceutical compositions of the present invention can be administered in various known ways, such as orally, parenterally, by inhalation spray, or via an implanted reservoir. The pharmaceutical compositions of the present invention can be administered alone or in combination with other anti-tumor drugs. Oral compositions can be any orally acceptable dosage form, including but not limited to tablets, capsules, emulsions, and suspensions, dispersions, and solutions. Commonly used pharmaceutically acceptable carriers or excipients include stabilizers, diluents, surfactants, lubricants, antioxidants, binders, colorants, fillers, emulsifiers, etc.

[0017] Sterile injectable compositions can be formulated using suitable dispersing or wetting agents and suspending agents according to techniques known in the art. Pharmaceutically acceptable carriers and solvents that can be used include water, mannitol, sodium chloride solution, etc.

[0018] The actual dosage level of the active ingredient in the pharmaceutical compositions of the present invention can be varied so as to obtain an amount of the active ingredient that is effective in achieving the desired therapeutic response for a particular patient, composition, and mode of administration and that is non-toxic to the patient. The selected dosage level depends on a variety of factors, including the activity of the specific compound or its salt of the present invention used, the route of administration, the time of administration, the excretion rate of the specific composition used, the duration of treatment, other drugs, compounds, and / or materials used in combination with the specific composition used, the age, sex, weight, general health, and medical history of the patient being treated, and similar factors well known in the medical arts.

[0019] Another object of the present invention is to provide the use of the compound of the present invention or its pharmaceutically acceptable salt, stereoisomer in the preparation of a drug for preventing and / or treating tumors.

[0020] The tumors include hepatocellular carcinoma, lung cancer, melanoma or colon cancer.

[0021] Beneficial effects:

[0022] The present invention synthesizes a small molecule compound called WX003. Pharmacological experiments have proven that this compound has good anti-tumor effects, can effectively inhibit the proliferation of various tumor cells, and has good prospects in the development of anti-tumor drugs. Description of the drawings

[0023] Figure 1 is the curve graph of the half maximal inhibitory concentration (IC 50 ) of WX003 against various tumor cell lines.

[0024] Figure 2 is the nuclear magnetic resonance hydrogen spectrum of WX003.

[0025] Figure 3 is the nuclear magnetic resonance carbon spectrum of WX003. Detailed implementation manners

[0026] The following describes the preparation method of WX003 of the present invention in combination with specific examples, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthesis methods described in this specification or known in the art, and such combinations can be easily carried out by those skilled in the art to which the present invention pertains.

[0027] The starting materials, reaction reagents, etc. used in the specific examples of the present invention are all commercially available. The present invention can be prepared in the form of salts by using common salt-forming methods in the art. For example, at room temperature, the compound is dissolved in hydrochloric acid ethanol for reaction to form hydrochloride; or benzenesulfonic acid is added thereto for reaction to form benzenesulfonate.

[0028] Example 1: Compound synthesis process and technology

[0029] One object of the present invention is to provide a preparation method of the compound of this patent, including the following steps:

[0030] Dissolve 6-hydroxy-2-naphthoic acid (31,500 mg, 2.66 mmol) in 2 ml of tetrahydrofuran (THF) for later use. Then dissolve phenethyl alcohol (325 mg, 2.66 mmol) and triphenylphosphine (TPP, 698 mg, 2.66 mmol) in 4 ml of tetrahydrofuran (THF). Under ice bath conditions, add diisopropyl azodicarboxylate (DIAD, 538 mg, 2.66 mmol) dropwise to the tetrahydrofuran solution of phenethyl alcohol and triphenylphosphine, stir well for 15 min, then add the previously prepared tetrahydrofuran solution of compound (31) dropwise to the reaction system in the ice bath, and transfer the system to room temperature and stir well. After monitoring by TLC that the raw materials have completely reacted, add water to quench the reaction. Extract with ethyl acetate three times, combine the organic phases, wash the organic phases with saturated sodium chloride solution, dry the obtained organic phases with anhydrous sodium sulfate, and then concentrate under reduced pressure. The crude product is purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1), and the obtained white solid is compound 32 with a yield of 82.3%.

[0031]

[0032] Dissolve the above compound 32 (254 mg, 0.87 mmol) in 2 ml of DMSO, then add 2-iodoxybenzoic acid (IBX, 269 mg, 0.96 mmol) to the reaction system, stir well, the reaction system changes from colorless to yellow and finally to orange-red. After monitoring by TLC that the raw materials have completely reacted, add water to quench the reaction. Extract with ethyl acetate three times, combine the organic phases, wash the organic phases with saturated sodium chloride solution, dry the obtained organic phases with anhydrous sodium sulfate, and then concentrate under reduced pressure to obtain an orange-red solid as compound 33 with a yield of 95%.

[0033] Dissolve the above compound 33 in 2 ml of acetonitrile (MeCN). Dissolve sodium dithionite (Na2S2O4, 166 mg, 0.96 mmol) in 2 ml of water. Slowly add the aqueous sodium dithionite solution to the acetonitrile solution of compound 33, and stir well under argon protection. The reaction system gradually changes from orange-red to light yellow. After monitoring by TLC that the reaction is complete, rotary evaporate under reduced pressure to remove acetonitrile, then extract with ethyl acetate three times, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, rotary dry to remove ethyl acetate, and then mix sodium dithionite with silica gel solid (1:8) to make sand. Column chromatograph the obtained sand sample with the above sodium dithionite and silica gel mixed solid (1:8) (petroleum ether: ethyl acetate = 4:1) to obtain a white solid as compound 29 with a yield of 70%.

[0034]

[0035] Compound 29 (310 mg, 1 mmol) was dissolved in dichloromethane. 7.5 ml of a mixed solution of sodium bicarbonate / potassium chloride (1 mol / L) was mixed with it, and then tetramethylammonium hydroxide (TDA, 500 mg, 1.5 mmol) and bromotetraacetylglucoside (600 mg, 1.5 mmol) were successively added to the reaction system and stirred well. After monitoring by TLC that the raw materials had completely reacted, extraction was carried out three times with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 4:1) to obtain compound 45 as a colorless oily liquid with a yield of 66%.

[0036]

[0037] The above-mentioned compound 45 was dissolved in 2 ml of methanol. A methanol solution of sodium methoxide (1 mol / L, 0.22 ml) was added to the aforementioned solution and stirred well. After monitoring by TLC that the raw materials had completely reacted, water and cation exchange resin were added to quench the reaction. Filtration was carried out by suction. After removing the cation exchange resin, the reaction solution was rotary evaporated under reduced pressure. The product was purified by silica gel column chromatography to obtain compound 41 as a white solid with a yield of 85%.

[0038]

[0039] Compound (41) White solid (44%); melting point (mp), 184 - 187 °C; 1 H NMR (300 MHz, Acetone-d 6 ) δ 8.52 (s, 1H), 8.26 (d, J = 8.8 Hz, 1H), 8.02 - 7.93 (m, 1H), 7.57 (t, J = 6.7 Hz, 2H), 7.46 - 7.31 (m, 4H), 7.26 (t, J = 7.1 Hz, 1H), 4.99 - 4.88 (m, 1H), 4.58 (t, J = 6.9 Hz, 2H), 4.04 - 3.75 (m, 2H), 3.71 - 3.47 (m, 4H), 3.16 (t, J = 6.9 Hz, 2H); 13 C NMR (75 MHz, Acetone-d 6 ) δ 166.04, 142.67, 142.10, 138.41, 130.48, 130.31, 129.10, 128.49, 127.25, 126.73, 126.48, 123.95, 122.28, 120.89, 120.51, 104.51, 77.32, 76.64, 73.97, 70.32, 65.41, 61.66, 34.97; MS (ESI) m / z, 472.16 [M + H]+ .

[0040] Example 2: Biological Activity Assay

[0041] Test method: Prepare a stock solution of WX003 at 100 mM.

[0042] For adherent cells Hep3B, Huh7, HepG2, B16-F10, H1299, H1975 and HCT-116, seed them into a 96-well plate at a density of 3000 cells / well with 100 μL per well. Prepare the corresponding small molecule stock solution as a drug-containing medium with a gradient dilution of 3-fold from 200 μM. After the cells adhered, discard the original medium, and add the gradient-diluted working solution to the wells seeded with cells at a volume of 100 μL per well. After culturing for 48 hours, measure the absorbance by the CCK8 method and calculate its IC 50 value.

[0043] The experimental results are shown in Table 1.

[0044] Table 1 IC 50 values (μM) of the anti-proliferative activities of the examples against 7 cancer cell lines

[0045] Compound Hep3B Huh7 HepG2 B16-F10 H1299 H1975 HCT-116 Example 22.54 15.92 12.88 15.88 13.40 17.60 17.56

[0046] Hep3B cells are a liver cancer cell line, widely used in the research of liver cancer; Huh7 is derived from a well-differentiated hepatocellular carcinoma of a Japanese male and is the only cell line that can effectively replicate hepatitis C virus; HepG2 cells are derived from the liver cancer tissue of a 15-year-old white girl and are suitable for the research on hepatocyte metabolism; B16-F10 cells are derived from the skin melanoma tissue of C57BL / 6J mice and are a sub-line of the B16 tumor cell line, widely used in the research of tumor cell formation, invasion and metastasis; H1299 cells are derived from the lymph node metastasis lesions of a 43-year-old male patient and are widely used in lung cancer research; H1975 cells are isolated from the non-small cell lung adenocarcinoma tissue of a female without a smoking history and are mainly used in lung cancer research and immuno-oncology research in scientific research; HCT-116 is isolated from the lymph nodes of a 51-year-old male colon cancer patient and is widely used in colorectal cancer and toxicology research.

[0047] Table 1 shows that WX003 can effectively inhibit the in vitro growth of hepatocellular carcinoma, lung cancer, colon cancer and melanoma cell lines.

[0048] The above experimental results show that the compound provided by the present invention can effectively inhibit the in vitro growth of hepatocellular carcinoma, lung cancer, colon cancer and melanoma cell lines.

[0049] The above-mentioned compounds of the present invention and their pharmaceutically acceptable salts can effectively inhibit the in vitro growth of hepatocellular carcinoma, lung cancer, colon cancer and melanoma cell lines, and can be used as active ingredients in pharmaceuticals. Therefore, a drug containing the above-mentioned compound as an active ingredient can be used to prepare a drug for preventing and / or treating tumors.

[0050] As described above, although the present invention has been shown and described with reference to specific preferred embodiments, it should not be construed as a limitation of the present invention itself. Various changes in form and detail may be made without departing from the spirit and scope of the present invention as defined by the appended claims.

Claims

1. A broad-spectrum anti-tumor compound or a pharmaceutically acceptable salt or stereoisomer thereof:

2. A method for preparing the compound according to claim 1.

3. Use of the compound according to claim 1 in the preparation of drugs for preventing and / or treating tumors.

4. The use according to claim 3, characterized in that: The tumor includes hepatocellular carcinoma, lung cancer, melanoma or colon cancer.

5. A pharmaceutical composition, characterized in that It comprises the compound as claimed in claim 1 and a pharmaceutically acceptable carrier or excipient.

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