Antitumor small molecule compounds, and methods of making and using the same

Abstract

The application relates to a kind of antitumor small molecule compounds and preparation method and purposes thereof.The application relates to an antitumor compound or its pharmaceutically acceptable salt, stereoisomer, pharmaceutical composition containing the same.The compound of the application is named as phenethyl 5-hydroxy-6-(((3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-2-naphthoate using IUPAC standard nomenclature, which has been proved to have good antitumor effect and can effectively inhibit the proliferation of various tumor cells.The application also relates to a preparation method of the compound and purposes in preparing a drug for preventing and / or treating tumors.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a compound with the function of inducing the death of multiple tumor cells, and a preparation method and use thereof. BACKGROUND

[0002] Tumor is an aggregation of abnormal cells. Compared with corresponding normal cells, tumor cells can achieve self-sufficiency of growth signals and are insensitive to growth inhibition signals in the body, showing unlimited growth ability and replication potential. The ability of tumor cells to promote peripheral angiogenesis, escape cell programmed death, and tissue invasion and metastasis also aggravates the degree of their malignant growth. At present, various tumors including liver cancer, lung cancer, melanoma and colon cancer have been widely characterized. Through existing tumor treatment methods, certain effects can be achieved in clinical practice and the survival time of patients can be prolonged. However, more than 9.7 million people died of tumors worldwide in 2022, and the incidence rate is increasing year by year, so tumor is still a global problem that seriously threatens human life and health.

[0003] Primary liver cancer (PLC) is a typical malignant tumor with the characteristics of 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 epidemiological survey results, hepatitis B virus (HBV) is a key factor leading to hepatocellular carcinoma, in addition to which hepatitis C virus (HCV), aflatoxin, alcohol, and nitrosamine compounds are also hepatocarcinogenic factors. At present, the main treatment methods for hepatocellular carcinoma include liver transplantation, liver resection, radiofrequency ablation, radiotherapy, and chemotherapy. When liver cancer progresses to the late stage, the targeted drugs represented by sorafenib and lenvatinib have the best efficacy, and in recent years, with the rise of immune checkpoint inhibitors and monoclonal antibodies, such as PD-L1 monoclonal antibody, progress has also been made in clinical practice. However, the first-line drugs for liver cancer at the present stage still have the limitations of limited selection, strong toxic and side effects, poor prognosis, and easy drug resistance. Therefore, there is an urgent need for effective new anti-tumor drugs or new drug combinations in clinical practice.

[0004] Lung cancer usually refers to primary bronchogenic carcinoma, which is the most common lung malignancy and the highest incidence of cancer in China, including small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). Non-small cell lung cancer accounts for 80% of total cases, with high mortality and strong heterogeneity. The incidence of lung cancer is closely related to smoking, air pollution, occupational exposure, genetic factors and other chronic lung diseases. According to the size of the primary tumor and whether lymph node metastasis and other organ invasion occur, lung cancer can be divided into stages I to IV according to TNM staging. For early and intermediate lung cancer patients, surgical treatment is the preferred option, and other treatment methods include chemoembolization and radiotherapy. According to whether the patient has key tumor target gene mutations such as EGFR, the main drug treatment for lung cancer is tyrosine kinase inhibitors such as gefitinib, erlotinib and osimertinib, which effectively prolongs the overall survival of patients. However, overall non-small cell lung cancer has poor sensitivity to chemotherapy and radiotherapy, and the development of new anticancer drugs remains a top priority in lung cancer treatment research.

[0005] Melanoma is a malignant tumor originating from melanocytes, which is the most dangerous skin cancer. It can be caused by the deterioration of existing pigmented nevi or new lesions. 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 spread. After spread, the survival period of melanoma patients will be significantly reduced, so it has the characteristics of high metastasis rate and high mortality. Epidemiological studies have shown that the incidence of melanoma is mainly in middle-aged and elderly men, with higher incidence and mortality in men than in women. The incidence of melanoma is mainly related to DNA damage caused by sunlight or ultraviolet light, genetic pathogenic genes, and local irritation such as repeated friction and trauma. For early patients, Mohs surgery and other surgeries can be used for complete cure. Drug treatment methods include chemotherapy with dacarbazine and paclitaxel, as well as targeted drug therapy for patients with BRAF and MEK mutations, including dabrafenib and trametinib. Treatment targeting a single target can promote drug resistance in melanoma, and the development of new types of anticancer compounds is expected to improve the drug resistance of melanoma and enhance the therapeutic effect.

[0006] Colorectal cancer (CRC), also known as colon cancer or rectal cancer, is a tumor that develops from the colon or rectum, and is the second highest incidence of cancer in China, with more than 500,000 patients, and the incidence is on the rise. Colon cancer has the characteristics of early symptoms not obvious, and the disease develops slowly, and most patients are in the middle and late stages when diagnosed, and therefore has the characteristics of limited treatment and poor prognosis. Its causes are related to age, genetic factors, chronic diseases such as ulcerative colitis, and are closely related to dietary habits such as alcohol abuse, high fat, high red meat and pickled food intake. The main diagnostic method for colon cancer is colonoscopy, which detects the extent of disease progression and whether it has spread through sampling and medical imaging. According to the diagnostic results, colon cancer can be treated by endoscopic surgery, chemotherapy such as capecitabine, fluorouracil and radiotherapy. According to the changes in epigenetics, targeted therapy such as cetuximab and bevacizumab can effectively prolong the survival of patients. However, the mechanism of colon cancer chemotherapy resistance is complex, and it is easy to cause treatment failure through KRAS mutation and signal bypass activation.

[0007] Therefore, it is of great significance to develop new spectral antitumor compounds for studying new drugs or pharmaceutical compositions for treating hepatocellular carcinoma and other cancers. SUMMARY

[0008] The purpose of the present application 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 referred to as WX003) or a pharmaceutically acceptable salt or stereoisomer thereof:

[0009]

[0010] In some preferred embodiments, the pharmaceutically acceptable salt includes an acid addition salt of the present compound with 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; and an acid salt of the present compound with an inorganic base.

[0011] In some more preferred embodiments, the pharmaceutically acceptable salt includes an alkali metal cation salt, an alkaline earth metal cation salt and an ammonium cation salt.

[0012] The compound involved in the present application can also exist in the form of a salt thereof, which is converted into the compound mentioned in the present patent in vivo. For example, within the scope of the present application, the compound of the present application is converted into a pharmaceutically acceptable salt form according to the process known in the art, and they are used in the form of a salt.

[0013] All tautomers of the compounds of the present application are included within the scope of the present application. The compounds of the present application can exist in particular geometric or stereoisomeric forms. The present application includes all such isomers, as well as mixtures thereof, including rac mixtures.

[0014] The compounds of the present application can be prepared by the above or similar methods, using appropriate starting materials according to the different substituents. Those skilled in the art will recognize that the above routes are useful for understanding the present application, but are not limiting of the present application, unless otherwise specified, the variables are defined as mentioned in general formula I.

[0015] Another object of the present application is to provide a pharmaceutical composition comprising a compound of the present application or a pharmaceutically acceptable salt, stereoisomer thereof, and a pharmaceutically acceptable carrier or excipient.

[0016] The pharmaceutical composition of the present application can be administered in a variety of ways, for example, orally, parenterally, by inhalation spray, or via an implanted reservoir. The pharmaceutical composition of the present application can be administered alone or in combination with other anti-tumor drugs. The oral composition 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, and the like.

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

[0018] The actual dose level of the active ingredients in the pharmaceutical composition of the present application can be varied to obtain an amount of the active ingredient that is effective for a particular patient, composition, and mode of administration, to achieve the desired therapeutic response, without being toxic to the patient. The selected dose level will depend on a variety of factors including the activity of the particular compound of the present application or salt thereof employed, the route of administration, the time of administration, the rate of excretion of the particular composition being employed, the duration of the treatment, other drugs, compounds, and / or materials used in combination with the particular composition employed, the age, sex, weight, general health condition, and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0019] Another object of the present application is to provide the use of a compound of the present application or a pharmaceutically acceptable salt, stereoisomer thereof, in the manufacture of a medicament for the prevention and / or treatment of tumors.

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

[0021] Beneficial effects:

[0022] This invention synthesizes a small molecule compound called WX003. Pharmacological experiments have demonstrated that this compound has good anti-tumor activity and can effectively inhibit the proliferation of various tumor cells, showing great promise in the development of anti-tumor drugs. Attached Figure Description

[0023] Figure 1 The half-maximal inhibitory concentration (IC50) of WX003 against various tumor cell lines. 50 (Curve graph)

[0024] Figure 2 This is the hydrogen NMR spectrum of WX003.

[0025] Figure 3 This is the carbon NMR spectrum of WX003. Detailed Implementation

[0026] The preparation method of WX003 of the present invention is described below with reference to specific embodiments, but these specific methods do not constitute any limitation on the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily performed by those skilled in the art.

[0027] The starting materials and reaction reagents used in the specific embodiments of this invention are all commercially available. This invention can be prepared into a salt form using methods commonly used in the art, such as: dissolving the compound in hydrochloric acid-ethanol at room temperature to generate hydrochloride; or adding benzenesulfonic acid to generate benzenesulfonate.

[0028] Example 1: Compound Synthesis Flow and Process

[0029] One object of this invention is to provide a method for preparing the compound of this patent, comprising the following steps:

[0030] 6-Hydroxy-2-naphthoic acid (31,500 mg, 2.66 mmol) was dissolved in 2 ml of tetrahydrofuran (THF) for later use. Phenylene alcohol (325 mg, 2.66 mmol) and triphenylphosphine (TPP, 698 mg, 2.66 mmol) were dissolved in 4 ml of tetrahydrofuran (THF). Diisopropyl azodicarbonate (DIAD, 538 mg, 2.66 mmol) was added dropwise to the tetrahydrofuran solution of phenylethanol and triphenylphosphine under ice bath conditions. The mixture was stirred thoroughly for 15 min. Then, the tetrahydrofuran solution of the initially prepared compound (31) was added dropwise to the reaction system in the ice bath, and the system was moved to room temperature and stirred thoroughly. After the reaction of the starting materials was complete as monitored by TLC, water was added to quench the reaction. The mixture was extracted three times with ethyl acetate, and the organic phases were combined. The organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 8:1), yielding a white solid, compound 32, in 82.3% yield.

[0031]

[0032] Compound 32 (254 mg, 0.87 mmol) was dissolved in 2 mL of DMSO. Then, 2-iodobenzoic acid (IBX, 269 mg, 0.96 mmol) was added to the reaction system and stirred thoroughly. The reaction system changed from colorless to yellow, and finally to orange-red. After the reaction was complete as monitored by TLC, water was added to quench the reaction. The mixture was extracted three times with ethyl acetate, and the organic phases were combined. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an orange-red solid, which was compound 33, in 95% yield.

[0033] Compound 33 was dissolved in 2 ml of acetonitrile (MeCN), and sodium dithionite (Na2S2O4, 166 mg, 0.96 mmol) was dissolved in 2 ml of water. The sodium dithionite aqueous solution was slowly added to the acetonitrile solution of compound 33, and the mixture was stirred thoroughly under argon protection. The reaction system gradually changed from orange-red to light yellow. After the reaction was complete as monitored by TLC, the acetonitrile was removed by rotary evaporation under reduced pressure, followed by extraction with ethyl acetate three times. The organic phases were combined and dried over anhydrous sodium sulfate. After removing the ethyl acetate by rotary evaporation, the organic phase was prepared into a slurry using a 1:8 mixture of sodium dithionite and silica gel. The obtained slurry was separated by column chromatography (petroleum ether: ethyl acetate = 4:1) using the aforementioned 1:8 mixture of sodium dithionite and silica gel to obtain a white solid, compound 29, with a yield of 70%.

[0034]

[0035] Compound 29 (310 mg, 1 mmol) was dissolved in dichloromethane and mixed with 7.5 mL of a sodium bicarbonate / potassium chloride mixed solution (1 mol / L). Tetramethylammonium hydroxide (TDA, 500 mg, 1.5 mmol) and bromotetraacetyl glucoside (600 mg, 1.5 mmol) were then added sequentially to the reaction system, and the mixture was stirred thoroughly. After the reaction was complete as monitored by TLC, the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give a colorless oily liquid, compound 45, in 66% yield.

[0036]

[0037] The above 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 solution. The mixture was stirred thoroughly. The reaction of the raw materials was monitored by TLC until it was complete. Water and cation exchange resin were added to quench the reaction. The mixture was filtered to remove the cation exchange resin. The reaction solution was then evaporated to dryness under reduced pressure. The product was purified by silica gel column chromatography and was a white solid, which was compound 41, with a yield of 85%.

[0038]

[0039] Compound (41) is a white solid (44%); melting point (mp), 184-187 °C. 1 H NMR(300MHz, Acetone-d6)δ8.52(s,1H),8.26(d,J=8.8Hz,1H),8.02-7.93(m,1H),7.57(t,J=6.7Hz,2H),7.46-7.31(m,4H), 7.26(t,J=7.1Hz,1H),4.99-4.88(m,1H),4.58(t,J=6.9Hz,2H),4.04-3.75(m,2H),3.71-3.47(m,4H),3.16(t,J=6.9Hz,2H); 13 C NMR (75MHz, Acetone-d6) δ166.04,142.67,142.10,138.41,130.48,130.31,129.10,128.49,127.25,126.73,126.48, 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: Assay for biological activity

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

[0042] For adherent cells Hep3B, Huh7, HepG2, B16-F10, H1299, H1975, and HCT-116, cells were spotted into 96-well plates at a density of 3000 cells / well and 100 μL per well. The corresponding small molecule stock solutions were prepared as 200 μM serially diluted 3-fold gradient drug-containing media. After cell adhesion, the original media was discarded, and 100 μL of the serially diluted working solution was added to each well of the culture plate. After 48 hours of incubation, absorbance was measured using the CCK8 assay, and IC50 was calculated using GraphpadPrism. 50 value.

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

[0044] Table 1. IC50 of the antiproliferative activity against 7 cancer cell lines in the examples. 50 Value (μM)

[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 liver cancer research; Huh7, derived from a well-differentiated hepatocellular carcinoma in a Japanese male, is the only cell line capable of effectively replicating the hepatitis C virus; HepG2 cells, derived from liver cancer tissue in a 15-year-old Caucasian girl, are suitable for research on hepatocyte metabolism; B16-F10 cells, derived from melanoma tissue in the skin of C57BL / 6J mice, are a subline of the B16 tumor cell line and are widely used to study tumor cell formation, invasion, and metastasis; H1299 cells, derived from lymph node metastases in a 43-year-old male patient, are widely used in lung cancer research; H1975 cells, isolated from non-small cell lung adenocarcinoma tissue in a non-smoking female, are mainly used in lung cancer and immuno-oncology research; HCT-116, isolated from lymph nodes in a 51-year-old male colon cancer patient, 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 compounds of the present invention and their medically 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, drugs containing the above compounds as active ingredients can be used to prepare drugs for the prevention and / or treatment of tumors.

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

Claims

1. A broad-spectrum antitumor compound or its pharmaceutically acceptable salt or stereoisomer: 。 2. Use of the compound according to claim 1 in the preparation of a drug for the prevention and / or treatment of tumors, wherein the tumor is selected from hepatocellular carcinoma, lung cancer, melanoma, or colon cancer.

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

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