阅读说明：本技术 一种化合物及其用途 (Compound and application thereof ) 是由 何达海 刘军 丁克毅 李丽梅 王晓玲 于 2021-08-18 设计创作，主要内容包括：本发明提供了一种化合物在制备治疗抗癌药物中的用途。本发明研究发现,上述化合物具备良好的抗癌活性,可将其作为新的具备抗癌活性的药物,为临床治疗癌症尤其是肺癌提供了一种新的可能。(The invention provides application of a compound in preparing a medicine for treating cancer. The research of the invention finds that the compound has good anticancer activity, can be used as a new medicine with anticancer activity, and provides a new possibility for clinically treating cancers, particularly lung cancer.)

1. The use of a compound of formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of cancer:

wherein R1 and R2 are respectively and independently selected from O, Me, H or nothing, and the two are not absent at the same time;

r3 is selected from H or OH;

r4 is selected from H or a glycosyl group.

2. Use according to claim 1, characterized in that: and when one of R1 and R2 is absent, the other is H.

3. Use according to claim 1, characterized in that: the compound is selected from the following structures:

4. use according to any one of claims 1 or 3, characterized in that: the glycosyl is selected from one of glucose, galactose, rhamnose, xylose and arabinose.

5. Use according to claim 4, characterized in that: the compound is selected from the following structures:

6. use according to any one of claims 1 to 5, characterized in that: the medicament is an anti-lung cancer medicament.

7. Use according to claim 6, characterized in that: the lung cancer is selected from non-small cell lung cancer.

8. Use according to claim 6, characterized in that: the lung cancer is selected from lung adenocarcinoma, lung squamous carcinoma and large cell carcinoma.

9. A compound having the formula:

10. a medicament, characterized by: comprising a compound according to claim 9.

Technical Field

The present invention relates to the field of natural phytochemistry.

Background

Cancer is a disease caused by the abnormal regulation and over proliferation of cells in the body. Hyperproliferative cells are called cancer cells, which often invade surrounding tissues and may even metastasize to other parts of the body via the systemic circulation and/or lymphatic system (cancer metastasis).

Lung cancer is one of the most rapidly growing malignancies that threaten human health and life. In many countries, the incidence and mortality of lung cancer have been reported to be significantly higher in recent 50 years, with lung cancer incidence and mortality in men accounting for the first of all malignancies, in women accounting for the second, and mortality accounting for the second. The etiology of lung cancer is not completely clear up to now, and a large amount of data show that a large amount of smoking for a long time has a very close relationship with the occurrence of lung cancer. Existing studies have demonstrated that: the probability of lung cancer of a large number of smokers in a long term is 10-20 times that of non-smokers, and the smaller the smoking starting age is, the higher the probability of lung cancer is. In addition, smoking not only directly affects the health of the user, but also has adverse effects on the health of surrounding people, so that the lung cancer prevalence of passive smokers is obviously increased. The incidence of lung cancer in urban residents is higher than that in rural areas, which may be related to urban atmospheric pollution and carcinogens contained in smoke dust. So no smoking should be advocated and the urban sanitation work should be enhanced.

Disclosure of Invention

The invention provides a medicine with anticancer activity, in particular to an application of a compound shown as a formula I or a pharmaceutically acceptable salt thereof in preparing a medicine for treating cancer:

wherein R1 and R2 are respectively and independently selected from O, Me, H or nothing, and the two are not absent at the same time;

r3 is selected from H or OH;

r4 is selected from H or a glycosyl group.

Wherein, when one of R1 and R2 is absent, the other is H.

Further, the compound is selected from the following structures:

wherein the glycosyl is selected from one of glucose, galactose, rhamnose, xylose and arabinose.

Still further, the compound is selected from the following structures:

wherein the drug is an anti-lung cancer drug, including non-small cell lung cancer (NSCLC) and Small Cell Lung Cancer (SCLC).

Further, the lung cancer is selected from non-small cell lung cancers, including but not limited to lung adenocarcinomas (e.g., acinar adenocarcinomas, papillary adenocarcinomas, bronchioloalveolar carcinomas (BACs), and solid adenocarcinomas and various mixed types), squamous lung cancers (e.g., papillary squamous cell carcinomas, clear cell squamous cell carcinomas, small cell squamous cell carcinomas, basal squamous cell carcinomas), large cell cancers (e.g., clear cell large cell carcinomas, basal cell-like large cell carcinomas, lymphoepitheliomatous carcinomas of the lung, large cell neuroendocrine carcinomas of the lung), and the like.

The invention also provides a compound, the structural formula of which is as follows:

based on the compound, the invention also provides a medicament, and the active ingredient of the medicament comprises the compound shown in the formula III.

The anticancer medicine of the present invention includes inhibiting the growth of cancer cell, promoting the apoptosis of cancer cell, etc.

The research of the invention finds that the compound has good anticancer activity, can be used as a new medicine with anticancer activity, and provides a new possibility for clinically treating cancers, particularly lung cancer.

The compound of the invention can be obtained by extracting and purifying tetrandra root or can be prepared by an organic synthesis method. For example, if the R4 site is H, i.e., the compound is an aglycone, it can be obtained by hydrolysis of the glycoside compound.

The Stephania tetrandra is dried root of Stephania tetrandra (Stephania tetrandra S.Moore) belonging to Stephania of Menispermaceae.

The medicine of the invention can also use pharmaceutically acceptable auxiliary materials or auxiliary components.

The term "pharmaceutically acceptable" is meant to include any material that does not interfere with the effectiveness of the biological activity of the active ingredient and is not toxic to the host to which it is administered.

The auxiliary materials are the general names of all the additional materials except the main medicine in the medicine preparation, and the auxiliary materials have the following properties: (1) no toxic effect on human body and few side effects; (2) the chemical property is stable and is not easily influenced by temperature, pH, storage time and the like; (3) has no incompatibility with the main drug, and does not influence the curative effect and quality inspection of the main drug; (4) does not interact with the packaging material.

The pharmaceutically acceptable auxiliary components have certain physiological activity, but the addition of the components does not change the dominance of the pharmaceutical composition in the process of treating diseases, but only plays an auxiliary effect, and the auxiliary effects are only the utilization of the known activity of the components and are auxiliary treatment modes which are commonly used in the field of medicine. If the auxiliary components are used in combination with the pharmaceutical composition of the present invention, the protection scope of the present invention should still be included.

Pharmaceutically acceptable adjuvants such as cellulose and its derivatives (e.g. sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g. stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g. soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g. propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g. tween), wetting agents (e.g. sodium lauryl sulfate), colorants, flavors, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

Wherein, the pharmaceutical composition is an oral administration preparation.

Of course, the mode of administration of the pharmaceutical composition of the present invention is not particularly limited, and topical administration may be used in addition to oral administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, granules and the like, and are not limited to the above dosage forms. In these solid dosage forms, the raw material or extract is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the material or extract may also be in microencapsulated form with one or more of the above excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the starting materials or extracts, the liquid dosage forms may contain inert diluents commonly employed in the art, such as water or other solvents, solubilizing agents and emulsifiers, e.g., ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide, and oils, especially cottonseed, groundnut, corn germ, olive, castor and sesame oils, or mixtures of such materials, and the like.

In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. In addition to the starting materials or extracts, the suspensions may contain suspending agents, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Dosage forms of the compositions of the present invention for topical administration include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable agents.

Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

The foregoing aspects of the present invention are explained in further detail below with reference to specific embodiments. It should not be understood that the scope of the above-described subject matter of the present invention is limited to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Drawings

FIG. 1: mass spectrum of compound 1

FIG. 2: process for preparation of Compound 1¹H NMR spectrum

FIG. 3: process for preparation of Compound 1¹³C NMR spectra

Detailed Description

Example 1

10kg of tetrandra root is crushed (5-10 meshes), and is leached at normal temperature for 24 hours by using 3.0L of 70% ethanol water (hydrochloric acid for adjusting the pH value to 3). And (3) performing suction filtration to obtain a total leaching solution, performing vacuum concentration to remove most of ethanol, adjusting the pH value to 9-10 with ammonia water, and extracting with chloroform to obtain about 90g of chloroform extract. The water phase part is loaded on a D101 type macroporous resin column, and is sequentially eluted by 30 percent and 70 percent ethanol water to respectively obtain about 60g and 30g of 30 percent and 70 percent ethanol water eluate.

The chloroform extract was partially applied to a silica gel column (Φ 12cm × 60cm), purified with chloroform-methanol 15: 1 to 5: gradient elution is carried out for 1, 1.5L of each gradient elution is carried out, one bottle of eluate is collected for each 150mL, the solvent is recovered by vacuum concentration, similar components are tracked and combined by a thin layer, and 10 components (sequentially ordered from STC-1 to STC-10) are obtained. Sample STC-1 was subjected to silica gel column chromatography (petroleum ether-ethyl acetate ═ 4: 1) to obtain 2(23.4mg), sample STC-2 was subjected to silica gel column chromatography (Φ 4cm × 45cm) to obtain compounds 3(14.1mg) and 4(18.3mg), sample STC-3 was subjected to silica gel column chromatography (Φ 4cm × 40cm) to obtain compounds 5(8.3mg) and 7(7.3mg), and sample STC-4 was subjected to silica gel column chromatography (petroleum ether-acetone ═ 1: 1) to obtain compounds 10(6.3g) and 11(8.6 g).

Loading 70% ethanol water eluate on Sephadex LH-20 (phi 4cm is multiplied by 90cm), eluting with methanol (collecting 1 bottle per 15mL, and naturally volatilizing solvent), tracking in thin layer, combining similar components to obtain five components (sequentially ordered as STC-701 to STC-705), eluting STC-702 with methanol (45%) -water (55%) (containing 0.1% ammonia water in water phase) on HPLC at constant rate of 35mL/min, and column specification (C18, phi 30mm is multiplied by 250mm) to obtain compound 6(t 18)_R＝23.7min,5.6mg)。

The 30% ethanol water eluate was applied to a reverse phase column (ODS-C18, 75 μm) and eluted with 15% and 75% methanol water, respectively, to give about 20g of STC301 and about 15g of STC302, respectively, the sample STC-301 was applied to LH-20 and eluted with methanol at a flow rate of about 1 drop/sec, 1 fraction was collected per 1/20 column volumes of the eluate, the similar components were spotted and combined to give STC-3011(17.1g), STC-3012(1.8g), the sample 3011 was applied to a silica gel column (Φ 8 cm. times.60 cm), the mixture was eluted with chloroform methanol 3: 1 isocratic elution, collecting one bottle per 150mL, vacuum concentrating to recover solvent, thin layer tracing, combining similar components to obtain 10 components (STC-3011-1 to STC-3011-10), subjecting sample STC-3011-6 to HPLC gradient elution, eluting with methanol-water 35% to 45% (0-50min), flowing at 35mL/min, and column specification (C18, phi 30mm × 250mm) to obtain compound 9 (t)_R(32.2 min,200mg) sample STC-3011-10 was eluted by HPLC gradient with methanol-water 5% to 15% (0-50min) flow rate of 35mL/min column format (C18, Φ 30mm × 250mm) to give compound 1(t 18)_R＝8min,5.2mg)、8(t_R＝31min,11.3g)。

Compound 1: light yellow powder is prepared from the raw materials of,λ_max＝282(0.938)，λ_max234(1.191), high resolution mass spectrum HR-ESI-MS excimer ion peak M/z:494.20273[ M + H: (M + H)]⁺(calculated 494.20262 for C₂₄H₃₂NO₁₀) Giving the formula C₂₄H₃₂NO₁₀。

¹H and ¹³C NMR(Table 1,400MHz for the ¹H NMR and 100MHz for the ¹³C NMR,CD₃OD)。

TABLE 1 NMR hydrogen and carbon spectra of Compound 1

The anticancer activity of each compound of the invention is researched (human lung cancer cell strain H1299, lung adenocarcinoma subtype), and the detection method comprises the following steps: part of the compounds were tested for IC50 activity on human lung cancer resistant cell line H1299 using the MTT method. The cell lines were cultured in DMEM medium (100U/mL penicillin, 100. mu.g/mL streptomycin) containing 10% fetal bovine serum at 37 ℃ in a 5% CO2 incubator.

The logarithmic phase cell line was inoculated into a 96-well plate at about 3000 cells/well and incubated at 37 ℃ in a constant temperature incubator for 24 hours. After 24h, the samples with different concentration gradients are respectively added into the culture wells, a blank control is set, and the cells are placed in a cell culture box for further culture for 72 h. mu.L of MTT medium (5mg/mL) was added to the medium and stained for 4 h. MTT solution is removed, 150 mu L DMSO is added into each hole, after shaking for 10min, the absorbance of each hole at the wavelength of 570nm is detected by a microplate reader, and the inhibition rate and IC50 are calculated.

TABLE 2

Gefitinib, an oral epidermal growth factor receptor tyrosine kinase (EGFR-TK) inhibitor (belonging to small molecule compound). Inhibition of EGFR-TK may block tumor growth, metastasis and angiogenesis, and increase apoptosis of tumor cells. Can be used for treating locally advanced or metastatic non-small cell lung cancer (NSCLC) which has been treated by chemotherapy.

The research of the invention shows that the compound can effectively resist cancers, particularly lung cancer cells; under the same test conditions, the IC50 of the compounds is lower than that of gefitinib, and partial compounds in the compounds are even significantly lower than that of gefitinib, such as compounds 1,3, 5, 7, 10 and the like. Indicating that the natural compound can effectively prevent or treat lung cancer, especially non-small cell lung cancer.