A flavonoid glycoside and its application in the preparation of anti-tumor drugs

Abstract

The present application provides a flavonoid glycoside and its application in the preparation of anti-tumor drugs. The structure of the flavonoid glycoside is shown in Formula I: The flavonoid glycoside compound has anti-tumor activity, has good inhibitory effects on non-small cell lung cancer, breast cancer and liver cancer, and is significantly superior to the positive control drug 5-fluorouracil, and has the potential to be prepared into anti-tumor drugs.

Description

Technical Field

[0001] The present application relates to the field of pharmaceutical technology, and specifically relates to a flavonoid glycoside and its application in the preparation of anti-tumor drugs. Background Art

[0002] Any discussion of the prior art throughout the specification should not be regarded as an admission that such prior art is well-known or constitutes a part of the common general knowledge in the art.

[0003] In recent years, the incidence and mortality of tumors have been on the rise. Malignant tumors are seriously threatening human health and life safety. It is estimated that more than 14 million people worldwide suffer from tumors every year, and currently tumors rank second among the global causes of death. Traditional tumor treatments use a combination of radiotherapy and chemotherapy, which cause greater damage to normal cells, and many adverse reactions occur in patients in the middle and late stages, leading to further aggravation of the condition. Therefore, it is particularly important to find new tumor treatment targets and develop new tumor treatment drugs. Compared with traditional drugs, small molecule targeted drugs have the advantages of smaller side effects, higher specificity, and higher drug efficacy, and are currently a research hotspot for anti-tumor drugs. Traditional Chinese medicine has a mild effect, good treatment effect, and few adverse reactions, and has obvious advantages in the treatment of malignant tumors. In recent years, extracting effective anti-tumor compounds from natural drugs is an important research direction for developing new anti-tumor drugs.

[0004] Flavonoid compounds are a class of natural products that are widely present in nature and have the characteristics of a wide range of physiological activities and low toxicity and side effects. In recent years, their anti-tumor effects have received extensive attention and research. The research results show that flavonoid compounds have significant prevention and treatment effects on a variety of common cancers such as lung cancer, breast cancer, colon cancer, prostate cancer, liver cancer, leukemia, ovarian cancer, gastric cancer, etc. The main mechanisms of flavonoid compounds against tumors are: antioxidant, anti-free radical, inducing apoptosis of tumor cells, affecting the cell cycle, regulating immunity, inhibiting tumor angiogenesis, inhibiting cyclooxygenase 2, inhibiting telomerase activity, etc. Summary of the Invention

[0005] The present invention provides a flavonoid glycoside compound, which has been tested to exhibit anti-tumor activity, has good inhibitory effects on cancer cells of non-small cell lung cancer, breast cancer, and liver cancer, and its effect is significantly better than that of the positive control drug 5-fluorouracil, and has the potential to be prepared into an anti-tumor drug.

[0006] Specifically, the present invention provides the following technical solutions.

[0007] In the first aspect of the present invention, the present invention provides a flavonoid glycoside compound, whose structure is shown in Formula I:

[0008]

[0009] Its chemical name is: 5-hydroxy-6-(naphthalen-1-ylmethoxy)-2-(4-(naphthalen-1-ylmethoxy)phenyl)-7-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-4H-chromen-4-one.

[0010] In a second aspect of the present invention, the present invention provides a pharmaceutical composition or pharmaceutical preparation, which comprises the flavonoid glycoside compound shown in Formula I; or further, further comprises at least one pharmaceutically acceptable excipient.

[0011] In some embodiments of the present invention, the pharmaceutical composition or pharmaceutical preparation uses the flavonoid glycoside compound shown in Formula I of the present invention as the active ingredient.

[0012] In some embodiments of the present invention, the pharmaceutical preparation can be administered via the gastrointestinal tract or non-gastrointestinal routes.

[0013] In some embodiments, the dosage forms for administration via the gastrointestinal tract include powders, tablets, granules, capsules, solutions, emulsions or suspensions; the non-gastrointestinal dosage forms include injection dosage forms, mucosal dosage forms and cavity dosage forms.

[0014] In some embodiments, the injection dosage forms include intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection and intracavitary injection; the mucosal dosage forms include eye drops, nasal drops, ophthalmic ointments, gargles, sublingual tablets, patches and film dressings; the cavity dosage forms include suppositories, aerosols, effervescent tablets, drops and dripping pills.

[0015] Some embodiments of the present invention include a method for producing the composition or pharmaceutical preparation, the method comprising mixing the active substance (at least comprising the compound shown in Formula I of the present invention) with suitable excipients, and preparing by any suitable method, usually by uniformly mixing the active substance with liquid and / or finely divided solid excipients in the required proportions, and then, if necessary, forming the resulting mixture into the desired shape. The excipients can play roles such as filling, binding, disintegrating and lubricating, and sometimes also play roles such as coloring, flavor correcting and beautifying. Commonly used excipients such as starches (such as corn starch, potato starch, etc.), sugars (such as glucose, sucrose, etc.), celluloses (such as microcrystalline cellulose, carboxymethyl cellulose, etc.) and inorganic salts (such as inorganic calcium salts, etc.), and more excipients can be as described in the eighth edition of the Handbook of Pharmaceutical Excipients (Paul J Sheskey, Walter G Cook, Colin G Cable).

[0016] In the third aspect of the present invention, the present invention provides the use of the flavonoid glycoside compound represented by Formula I or a composition or pharmaceutical preparation containing the compound in the preparation of an anti-tumor drug.

[0017] In some embodiments of the present invention, the tumors include but are not limited to non-small cell lung cancer, breast cancer, and liver cancer.

[0018] In the fourth aspect of the present invention, the present invention provides a method, which includes administering an effective dose of the flavonoid glycoside compound represented by Formula I or a composition or pharmaceutical preparation containing the compound to a subject for treating cancer.

[0019] In some embodiments of the present invention, the cancers include but are not limited to non-small cell lung cancer, breast cancer, and liver cancer.

[0020] The subject refers to an animal that has already been the object of treatment, observation, or experiment, preferably a mammal, and most preferably a human.

[0021] The effective dose refers to the amount of the active compound or composition or pharmaceutical preparation including the flavonoid glycoside compound represented by Formula I of the present invention, and this amount can cause the biological or medical response of the tissue system, animal, or human pursued by researchers, veterinarians, doctors, or other medical personnel, which includes alleviating or partially alleviating the symptoms of the treated disease, syndrome, disorder, or condition.

[0022] In some embodiments of the present invention, the method can be used in combination with other anti-tumor drugs or in combination with other anti-tumor therapies.

[0023] Compared with the prior art, the advantages of the present invention include:

[0024] The present invention provides a flavonoid glycoside compound, and its structure is as shown in Formula I. This compound has been tested to have anti-tumor activity, shows good inhibitory effects on the cancer cells of non-small cell lung cancer, breast cancer, and liver cancer, and its effect is significantly better than that of the positive control drug 5-fluorouracil, and it has the potential to be prepared into an anti-tumor drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The specification drawings forming a part of this application are used to provide a further understanding of this application. The schematic embodiments and descriptions thereof of this application are used to explain this application and do not constitute an improper limitation of this application. Hereinafter, the embodiments of this application will be described in detail in conjunction with the drawings, where:

[0026] Figure 1 is the 1 H-NMR of the flavonoid glycoside shown in Formula I in the preparation example of the present invention.

[0027] Figure 2For the flavonoid glycoside shown in Formula I in the embodiments of the present invention 13 C-NMR. Specific embodiments

[0028] The following further elaborates on the present application in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present application and not to limit the scope of the present application. For the experimental methods without specific conditions indicated in the following embodiments, they are generally carried out under conventional conditions or according to the conditions recommended by the manufacturer.

[0029] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. The reagents or raw materials used in the present application can all be obtained through conventional channels. Unless otherwise specified, the reagents or raw materials used in the present application are used in the conventional manner in the art or according to the product instructions. In addition, any methods and materials similar or equivalent to the described content can be applied to the methods of the present application. The preferred implementation methods and materials described in the text are only for demonstration purposes.

[0030] Preparation Example Preparation of flavonoid glycoside

[0031] The structure of the flavonoid glycoside is as shown in Formula I, and its preparation route is as follows:

[0032]

[0033] 200 mL of anhydrous methanol was added to a 500 mL round-bottom flask, and SOCl 2 (7.25 mL, 100 mmol) was slowly added dropwise under an ice bath. After the addition was completed, the ice bath was removed, and stirring was continued at room temperature for 1 h. Then, scutellarin (Compound 1, 4.62 g, 10 mmol) was added, and after stirring at room temperature for 9 h, TLC (V 乙酸乙酯 :V 异丙醇 :V 水 =4:2:1) was used to detect that the reaction was complete. It was directly filtered to obtain scutellarin methyl ester (Compound 2, 4.67 g, 98%); 1 H-NMR (400 MHz, DMSO-d 6 ) δ 12.85 (s, 1H), 10.39 (s, 1H), 7.93 (d, J = 9.0 Hz, 2H), 7.00 (s, 1H), 6.94 (d, J = 9.0 Hz, 2H), 6.81 (s, 1H), 5.28 (d, J = 7.0 Hz, 1H), 4.20 (d, J = 6.0 Hz, 1H), 3.70 - 3.90 (m, 3H), 3.68 (s, 3H); 13 C-NMR (125 MHz, DMSO-d 6): δ 182.8, 169.7, 164.5, 161.6, 151.3, 149.4, 147.3, 130.8, 128.9, 121.7, 116.4, 106.3, 102.9, 100.2, 93.9, 75.7, 75.4, 73.1, 71.8, 52.4 (-OCH 3 )。

[0034] In a 50 mL round-bottom flask, compound 2 (1.19 g, 2.50 mmol) was dissolved in 25 mL of DMF. Under stirring at room temperature, DIPEA (2.20 mL, 12.50 mmol) and NapBr (1.39 g, 6.25 mmol) were added successively. After stirring overnight at room temperature, TLC (V 二氯甲烷 :V 甲醇 = 20:1) was used to detect the disappearance of the starting materials. The reaction mixture was poured into 100 mL of ice water, extracted with ethyl acetate (40 mL × 3), washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by dry-column chromatography to obtain compound 3 (1.65 g, 87%); 1 1H NMR (400 MHz, DMSO-d 6 ) δ 13.01 (s, 1H), 8.04 (t, J = 8.7 Hz, 4H), 7.98–7.88 (m, 6H), 7.75 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 8.4 Hz, 1H), 7.52 (dd, J = 9.1, 6.7 Hz, 4H), 7.26 (d, J = 8.7 Hz, 2H), 7.15 (s, 1H), 6.94 (s, 1H), 5.75 (d, J = 5.2 Hz, 1H), 5.56 (d, J = 5.3 Hz, 1H), 5.47 (d, J = 7.5 Hz, 1H), 5.45–5.37 (m, 3H), 5.28 (d, J = 11.2 Hz, 1H), 5.14 (d, J = 11.2 Hz, 1H), 4.26 (d, J = 9.5 Hz, 1H), 3.66 (s, 3H), 3.46 (ddd, J = 14.0, 10.6, 5.7 Hz, 3H); 13 13C-NMR (125 MHz, DMSO-d 6): δ 182.37, 169.24, 163.86, 161.56, 156.12, 152.87, 152.27, 135.21, 134.09, 132.77, 132.72, 132.65, 132.62, 131.32, 128.43, 128.19, 127.90, 127.83, 127.68, 127.64, 127.56, 126.85, 126.55, 126.41, 126.27, 126.12, 126.06, 125.74, 123.00, 115.52, 105.94, 103.51, 99.33, 93.88, 75.77, 75.34, 74.25, 72.98, 71.37, 69.73, 52.02。

[0035] Compound 3 (0.80 g, 1.06 mmol) was suspended in 30 mL of methanol, and sodium borohydride (0.80 g, 21.20 mmol) was added in small portions and multiple times under vigorous stirring at room temperature. After 3 h, TLC (V 二氯甲烷 ∶V 甲醇 = 15:1) detection showed that the reaction was complete. 15 mL of 10% acetic acid solution was added to the reaction system, and it was extracted with ethyl acetate (50 mL × 3). The organic phase was concentrated under reduced pressure to obtain the crude product. The obtained crude product was suspended in 10 mL of ethyl acetate, heated under reflux for 30 min, then cooled naturally, filtered, and the filter cake was washed with ethyl acetate and dried in vacuo to obtain the light yellow solid flavone glycoside of formula I (0.65 g, 85%). 1 1H-NMR (400 MHz, DMSO-d 6 ) δ 13.00 (s, 1H), 8.17–7.85 (m, 10H), 7.76 (d, J = 7.3 Hz, 1H), 7.60 (d, J = 7.7 Hz, 1H), 7.52 (s, 4H), 7.25 (d, J = 7.5 Hz, 2H), 7.09 (s, 1H), 6.95 (s, 1H), 5.60 (s, 1H), 5.40 (s, 2H), 5.22 (dt, J = 36.7, 14.3 Hz, 5H), 4.68 (s, 1H), 3.74 (s, 1H); 13 13C-NMR (125 MHz, DMSO-d 6): δ 182.42, 163.88, 161.60, 156.70, 152.71, 152.35, 135.34, 134.15, 132.82, 132.78, 132.70, 132.67, 131.39, 128.51, 128.25, 127.97, 127.89, 127.72, 127.61, 126.94, 126.65, 126.59, 126.48, 126.34, 126.18, 126.11, 125.78, 123.04, 115.58, 105.82, 103.50, 100.17, 94.40, 77.34, 76.88, 74.30, 73.42, 69.76, 69.65, 60.71, 54.95. The hydrogen spectrum and carbon spectrum of the flavonoid glycoside shown in Formula I are respectively as Figure 1 and Figure 2 shown in.

[0036] Experimental Example Antitumor Activity Test of Flavonoid Glycoside

[0037] Test cells: Three different types of human tumor cells were used in the experiment, including human non-small cell lung cancer cell A549, human breast cancer cell MCF-7, and liver cancer cell HepG 2.

[0038] Test method:

[0039] 1. IC 50 Determination

[0040] 1) Drug preparation: The compound of Formula I (Preparation Example) was dissolved in DMSO to a concentration of 10 mM, and then serially diluted 10-fold to 1 mM, 100 μM, 10 μM, 1 μM, and 0.1 μM. The stock solution concentration and working concentration are shown in the following table:

[0041] Stock solution concentration Working concentration 10 mM 100 μM 1 mM 10 μM 100 μM 1 μM 10 μM 100 nM 1 μM 10 nM 0.1 μM 1 nM

[0042] 2) Cell culture and plating: All three types of cells were cultured in DMEM medium containing 10% FBS. When the confluence reached 75 - 85%, they were digested and counted, and then evenly plated into 96-well plates. The initial cell numbers are shown in the following table:

[0043] Cell Number of cells per well A549 2000 MCF-7 3000 HepG 2 2000

[0044] 3) Drug addition: The drugs were added 24 hours after cell plating, and each plate was set with a DMSO solvent control well.

[0045] 4) MTT assay: After 48 hours of drug addition, MTT was added. After incubation for 4 hours, the supernatant was discarded, 100 μL of DMSO was added, and the plate was shaken on a shaker for 10 min and then detected for OD 570 and OD 720For the numerical value, the calculation formula for cell viability is as follows:

[0046] Cell viability = (drug-treated group OD570 - drug-treated group OD720 ) / (DMSO group OD570 - DMSO group OD720 ) × 100%

[0047] The IC of the compound was calculated by Graphpad software 50 .

[0048] 2. Test results:

[0049] IC 50 Detection results (unit: μM)

[0050]

[0051]

[0052] From the data analysis in the above table, it can be seen that the flavonoid glycoside compound shown in Formula I shows good inhibitory activity against human non-small cell lung cancer cell line A549, human breast cancer cell line MCF-7, and liver cancer cell line HepG 2. Its tumor inhibitory activity is significantly better than that of the positive control drug 5-fluorouracil, indicating that this flavonoid glycoside has good anti-tumor application prospects.

[0053] The above are only the preferred embodiments of the present application and are not used to limit the present application. Although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present application shall be included within the protection scope of the present application.

Claims

1. Use of a compound represented by formula I, or a composition or pharmaceutical preparation containing the compound, in the preparation of an anti-tumor drug; The tumor is non-small cell lung cancer, breast cancer or liver cancer.

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