阅读说明：本技术 白花蛇舌草抗肿瘤有效组分及其制备方法和用途 (Oldenlandia diffusa anti-tumor effective component and preparation method and application thereof ) 是由 王英锋 洪雅 于 2020-04-02 设计创作，主要内容包括：本发明涉及医药领域,具体涉及一种白花蛇舌草抗肿瘤有效组分及其制备方法和用途。本发明提供的一种山奈酚-3-O-[2-O-(6-O-E-阿魏酰基)-β-D-吡喃葡萄糖基]-β-D-吡喃半糖和E-6-O-对香豆酰鸡屎藤苷甲酯单独或组合在抗肿瘤或者制备抗肿瘤药物中的用途,本发明研究发现山奈酚-3-O-[2-O-(6-O-E-阿魏酰基)-β-D-吡喃葡萄糖基]-β-D-吡喃半糖化合物单体和E-6-O-对香豆酰鸡屎藤苷甲酯化合物单体各自都具有很强的抗肿瘤活性,在体内抗肿瘤活性研究中,抑制率可以分别达到48.97％和45.96％,而当两者组合抑制肿瘤细胞时,发现两者具有显著的协同抑制肿瘤的作用。(The invention relates to the field of medicines, and in particular relates to an oldenlandia antitumor active component, and a preparation method and application thereof. The invention provides an application of kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-glucopyranose and E-6-O-p-coumaroyl paederoside methyl ester in anti-tumor or anti-tumor drug preparation, the invention researches to find that kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-glucopyranose compound monomer and E-6-O-p-coumaroyl paederoside methyl ester compound monomer respectively have strong anti-tumor activity, in the research of in vivo anti-tumor activity, the inhibition rate can reach 48.97% and 45.96% respectively, and when the two are combined to inhibit tumor cells, the two are found to have obvious synergistic tumor inhibition effect.)

1. An application of kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-glucopyranose and E-6-O-p-coumaroyl paederoside methyl ester in antitumor or antitumor drug preparation is provided.

2. The use of claim 1, wherein said tumor comprises liver cancer.

3. Use according to claim 1 or 2, characterized in that the mass ratio of kaempferol-3-O- [2-O- (6-O-E-feruloyl) - β -D-glucopyranosyl ] - β -D-pyranose hemi-sugar and E-6-O-p-coumaroyl paederosidin methyl ester is 1:1-1:5 or 2:1-5:1 when combined.

4. Use according to claim 1 or 2, characterized in that, when combined, the mass ratio of kaempferol-3-O- [2-O- (6-O-E-feruloyl) - β -D-glucopyranosyl ] - β -D-pyranose semi-saccharide and E-6-O-p-coumaroyl paederoside methyl ester is 1:2-1:5 or 2:1-5: 1; preferably, the mass ratio is 1:2, 1:3, 1:4, 1:5, 2:1, 3:1, 4:1 or 5: 1.

5. Use according to claim 1 or 2, characterized in that the mass ratio of kaempferol-3-O- [2-O- (6-O-E-feruloyl) - β -D-glucopyranosyl ] - β -D-pyranose semi-saccharide and E-6-O-p-coumaroyl paederosidin methyl ester, when combined, is 3: 1.

6. An anti-tumor effective component of oldenlandia diffusa, which is characterized by comprising kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranosyl and/or E-6-O-p-coumaroyl paederoside methyl ester.

7. The oldenlandia diffusa antitumor active ingredient according to claim 6, wherein the mass ratio of kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranosyl and E-6-O-p-coumaroyl paederoside methyl ester is 1:1-1:5 or 2:1-5:1 when combined.

8. The oldenlandia diffusa antitumor active ingredient according to claim 6 or 7, wherein in combination, the mass ratio of kaempferol-3-O- [2-O- (6-O-E-feruloyl) - β -D-glucopyranosyl ] - β -D-pyranosyl and E-6-O-p-coumaroyl paederoside methyl ester is 1:2-1:5 or 2:1-5: 1; preferably, the mass ratio is 1:2, 1:3, 1:4, 1:5, 2:1, 3:1, 4:1 or 5: 1.

9. The oldenlandia diffusa antitumor active ingredient according to claim 6 or 7, wherein the mass ratio of kaempferol-3-O- [2-O- (6-O-E-feruloyl) - β -D-glucopyranosyl ] - β -D-pyranosyl and E-6-O-p-coumaroyl paederoside methyl ester is 3:1 when combined.

10. The oldenlandia diffusa antitumor active ingredient according to claim 6 or 7, wherein the preparation method of the oldenlandia diffusa antitumor active ingredient comprises:

dissolving the extract of oldenlandia diffusa in 3-7% alcohol solution by volume percentage, loading macroporous resin, statically adsorbing, then sequentially eluting with 8-12%, 18-22%, 28-32% and 38-42% alcohol solution by volume percentage, collecting 38-42% alcohol solution eluent, and concentrating to obtain the effective part of oldenlandia diffusa;

dissolving the obtained effective part of the oldenlandia diffusa into 15-25% of alcohol solution by volume percentage, separating by adopting a gel reduced pressure chromatography column, eluting by using the alcohol solution with the volume percentage of 15-25%, collecting eluent to obtain eluent containing E-6-O-p-coumaroyl paederoside methyl ester, then eluting by using the alcohol solution with the volume percentage of 45-55%, collecting the eluent to obtain solution containing kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose.

11. The effective component of oldenlandia diffusa for resisting tumor according to claim 10, wherein the preparation method of the effective component of oldenlandia diffusa for resisting tumor comprises:

dissolving the extract of oldenlandia diffusa in 5% ethanol solution by volume percentage, loading macroporous resin, statically adsorbing, then sequentially eluting with 10%, 20%, 30% and 40% ethanol solution by volume percentage, collecting the eluent of 40% ethanol solution by volume percentage, and concentrating to obtain the effective part of oldenlandia diffusa;

dissolving the obtained effective part of the oldenlandia diffusa into an ethanol solution with the volume percentage of 20%, separating by adopting a gel reduced pressure chromatography column, eluting by using the ethanol solution with the volume percentage of 20%, collecting eluent to obtain eluent containing the E-6-O-p-coumaroyl paederoside methyl ester, then eluting by using the ethanol solution with the volume percentage of 50%, and collecting the eluent to obtain a solution containing the kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose.

12. An antitumor pharmaceutical composition comprising the oldenlandia diffusa antitumor active ingredient according to any one of claims 6 to 11.

13. The antineoplastic pharmaceutical composition of claim 12, further comprising a pharmaceutically acceptable excipient or carrier.

14. The antitumor pharmaceutical composition as claimed in claim 13, wherein the formulation form of the antitumor pharmaceutical composition comprises a liquid formulation and a solid formulation; preferably, the preparation form comprises injection, infusion solution, medicinal granules, powder, oral liquid, spray, powder injection, granules, tablets, sugar-coated tablets, film-coated tablets, enteric-coated tablets, orally disintegrating tablets, capsules, hard capsules, soft capsules, buccal tablets, granules, pills, dripping pills, pellets, paste, pellets or disintegrating agents.

Technical Field

The invention relates to the field of medicines, and in particular relates to an oldenlandia antitumor active component, and a preparation method and application thereof.

Background

Oldenlandia diffusa (Hedyotis diffusa Willd.) is a plant of the genus rubiaceae and is widely distributed in subtropical regions. The plant grows in Yunnan, Guangxi, Guangdong, Fujian, Zhejiang, Jiangsu, Anhui and the like. The main effects of the oldenlandia diffusa are heat clearing, detoxifying and diuresis, and pharmacological studies show that the oldenlandia diffusa has the effects of resisting bacteria, enhancing the immune function, resisting tumors, resisting aging and the like. The oldenlandia diffusa is complex in components and mainly contains anthraquinone, terpenoid, flavonoid, sterol, organic acid, polysaccharide, alkaloid, trace elements, amino acid and volatile components. The anthraquinone component mainly contains alizarin type, the flavonoid component mainly contains quercetin, kaempferol and the like, the terpene component mainly contains iridoid, the sterol compound mainly contains beta-sitosterol and stigmasterol, and the organic acid mainly contains ursolic acid, oleanolic acid and ferulic acid.

Malignant tumor seriously threatens human life health and is the disease with the highest death rate all over the world. With the change of life style and the increase of environmental pollution, the incidence rate of the disease tends to rise year by year in recent years. Oldenlandia diffusa contains rich antitumor active ingredients, and the ingredients with antitumor activity reported at present mainly comprise anthraquinones, terpenoids, flavonoids, sterols, organic acids, polysaccharides and volatile ingredients. The anti-tumor active substances are separated and extracted from the oldenlandia diffusa, and the oldenlandia diffusa is developed into a new medicine with anti-tumor curative effect, and has important application value and wide development prospect. Therefore, on the basis of the previous research, the invention extracts the effective part of the oldenlandia diffusa, and separates the effective anti-tumor component of the oldenlandia diffusa through a large amount of research, thereby having very important significance for developing new anti-tumor medicines.

Disclosure of Invention

Therefore, the technical problem to be solved by the invention is to provide an oldenlandia diffusa antitumor active component, and a preparation method and application thereof.

Therefore, the invention provides the following technical scheme:

in a first aspect, the invention provides an application of kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose semi-sugar and E-6-O-p-coumaroyl paederoside methyl ester in antitumor or antitumor drug preparation.

Preferably, the tumor comprises liver cancer.

Preferably, when combined, the mass ratio of kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose semi-sugar to E-6-O-p-coumaroyl paederoside methyl ester is 1:1-1:5 or 2:1-5: 1.

Preferably, when combined, the mass ratio of kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose semi-sugar to E-6-O-p-coumaroyl paederosidin methyl ester is 1:2-1:5 or 2:1-5: 1; preferably, the mass ratio is 1:2, 1:3, 1:4, 1:5, 2:1, 3:1, 4:1 or 5: 1.

Preferably, when combined, the mass ratio of kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose semi-sugar to E-6-O-p-coumaroyl paederoside methyl ester is 3: 1.

In a second aspect, the invention provides an effective component of oldenlandia diffusa for resisting tumor, which comprises kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose semi-sugar and E-6-O-p-coumaroyl paederoside methyl ester.

Preferably, the mass ratio of the kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose semi-sugar to the E-6-O-p-coumaroyl paederoside methyl ester is 1:1-1:5 or 2:1-5: 1.

Preferably, the mass ratio of the kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose semi-sugar to the E-6-O-p-coumaroyl paederoside methyl ester is 1:2-1:5 or 2:1-5: 1; preferably, the mass ratio is 1:2, 1:3, 1:4, 1:5, 2:1, 3:1, 4:1 or 5: 1.

Preferably, the mass ratio of the kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose semi-sugar to the E-6-O-p-coumaroyl paederoside methyl ester is 3: 1.

Preferably, the preparation method of the oldenlandia diffusa antitumor active ingredient comprises the following steps:

the preparation method of the coumaroyl paederoside methyl ester comprises the following steps:

dissolving the extract of oldenlandia diffusa in 3-7% alcohol solution by volume percentage, loading macroporous resin, statically adsorbing, then sequentially eluting with 8-12%, 18-22%, 28-32% and 38-42% alcohol solution by volume percentage, collecting 38-42% alcohol solution eluent, and concentrating to obtain the effective part of oldenlandia diffusa;

dissolving the obtained effective part of the oldenlandia diffusa into 15-25% of alcohol solution by volume percentage, separating by adopting a gel reduced pressure chromatography column, eluting by using the alcohol solution with the volume percentage of 15-25%, collecting eluent to obtain eluent containing E-6-O-p-coumaroyl paederoside methyl ester, then eluting by using the alcohol solution with the volume percentage of 45-55%, collecting the eluent to obtain solution containing kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose.

Preferably, the preparation method of the oldenlandia diffusa antitumor active ingredient comprises the following steps:

dissolving the extract of oldenlandia diffusa in 5% ethanol solution by volume percentage, loading macroporous resin, statically adsorbing, then sequentially eluting with 10%, 20%, 30% and 40% ethanol solution by volume percentage, collecting the eluent of 40% ethanol solution by volume percentage, and concentrating to obtain the effective part of oldenlandia diffusa;

dissolving the obtained effective part of the oldenlandia diffusa into an ethanol solution with the volume percentage of 20%, separating by adopting a gel reduced pressure chromatography column, eluting by using the ethanol solution with the volume percentage of 20%, collecting eluent to obtain eluent containing the E-6-O-p-coumaroyl paederoside methyl ester, then eluting by using the ethanol solution with the volume percentage of 50%, and collecting the eluent to obtain a solution containing the kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose.

In a third aspect, the invention provides an anti-tumor pharmaceutical composition, which comprises the oldenlandia diffusa anti-tumor effective component.

Preferably, the composition also comprises pharmaceutically acceptable excipients or carriers.

Preferably, the preparation forms of the antitumor drug composition comprise a liquid preparation and a solid preparation; preferably, the preparation form comprises injection, infusion solution, medicinal granules, powder, oral liquid, spray, powder injection, granules, tablets, sugar-coated tablets, film-coated tablets, enteric-coated tablets, orally disintegrating tablets, capsules, hard capsules, soft capsules, buccal tablets, granules, pills, dripping pills, pellets, paste, pellets or disintegrating agents.

The technical scheme of the invention has the following advantages:

1. the invention provides an application of kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-glucopyranose and E-6-O-p-coumaroyl paederoside methyl ester in anti-tumor or anti-tumor drug preparation, the invention researches to find that kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-glucopyranose compound monomer and E-6-O-p-coumaroyl paederoside methyl ester compound monomer have strong anti-tumor activity respectively, in the research of in vivo antitumor activity, the dose of E-6-O-p-coumaroyl paederoside methyl ester is 5mg/kg, the inhibition rate can reach 48.97%, the inhibition rate of kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-glucopyranose also reaches 45.96%, and when the two are combined to inhibit tumor cells, the two have obvious synergistic tumor inhibition effect.

2. The invention provides an anti-tumor effective component of oldenlandia diffusa, which comprises kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-pyranose semi-sugar and E-6-O-p-coumaroyl paederoside methyl ester; the anti-tumor effective component of the oldenlandia diffusa has obvious tumor inhibiting effect, and researches show that the kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-glucopyranose and E-6-O-p-coumaroyl paederoside methyl ester have obvious anti-tumor effect when the mass ratio of the kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-glucopyranose and E-6-O-p-coumaroyl paederoside methyl ester is 1:1 or 2:1-5:1, and when the mass ratio of the kaempferol-3-O- [2-O- (6-O-E-feruloyl) -beta-D-glucopyranosyl ] -beta-D-glucopyranose and E-6-O-p-coum, the antitumor effect is highest.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a liquid chromatogram of monomer C in example 1 of the present invention;

FIG. 2 is a mass spectrum of monomer C in example 1 of the present invention;

FIG. 3 is a liquid chromatogram of the F monomer species of example 1 of the present invention;

FIG. 4 is a mass spectrum of the F monomer in example 1 of the present invention;

FIG. 5 is a graph showing the results of the relative spleen weight of a mouse in an in vivo antitumor test after the mouse is inoculated with a tumor in Experimental example 1 of the present invention;

FIG. 6 is a graph showing the results of the weights of the thymus glands of mice in an in vivo antitumor test after the mice were inoculated with tumors in Experimental example 1 of the present invention;

FIG. 7 is a graph showing the results of comparison of the number of leukocytes in mice in an in vivo antitumor test after inoculation of tumors in mice in Experimental example 1 of the present invention;

FIG. 8 is a graph showing the results of the percentage of mouse lymphocyte counts in the in vivo antitumor test after the mice were inoculated with tumors in Experimental example 1 of the present invention.

Detailed Description

The following examples are provided to further understand the present invention, not to limit the scope of the present invention, but to provide the best mode, not to limit the content and the protection scope of the present invention, and any product similar or similar to the present invention, which is obtained by combining the present invention with other prior art features, falls within the protection scope of the present invention.