AU2021100051A4

AU2021100051A4 - Antitumor formula and extraction method thereof

Info

Publication number: AU2021100051A4
Application number: AU2021100051A
Authority: AU
Inventors: Yingjie Wang
Original assignee: Individual
Current assignee: Individual
Priority date: 2021-01-06
Filing date: 2021-01-06
Publication date: 2021-03-25
Anticipated expiration: 2029-01-06

Abstract

Austracy The present invention provides an antitumor formula which includes the following raw materials in parts by weight: 10-20 parts of nucleic acids, 10-20 parts of lectin, 10-20 parts of polysaccharides, 10-20 parts of ginsenosides, 5-10 parts of lipids and 5-10 parts of sterols. The antitumor formula in the present invention is in conformity with the "assistant and guide" principle required in traditional Chinese medicine prescriptions and compositions. The antitumor formula is precise in composition and appropriate in compatibility and is in conformity with therapeutic principles of "weakening induration", "eliminating stagnation", "treating retention with purgation" and "tonifying deficiency" during tumor treatment of the traditional Chinese medicine. With the adoption of various raw materials in the present invention, an effect of effectively treating tumors can be achieved. 1

Description

Austracy

The present invention provides an antitumor formula which includes the following raw materials in parts by weight: 10-20 parts of nucleic acids, 10-20 parts of lectin, 10-20 parts of polysaccharides, 10-20 parts of ginsenosides, 5-10 parts of lipids and 5-10 parts of sterols. The antitumor formula in the present invention is in conformity with the "assistant and guide" principle required in traditional Chinese medicine prescriptions and compositions. The antitumor formula is precise in composition and appropriate in compatibility and is in conformity with therapeutic principles of "weakening induration", "eliminating stagnation", "treating retention with purgation" and "tonifying deficiency" during tumor treatment of the traditional Chinese medicine. With the adoption of various raw materials in the present invention, an effect of effectively treating tumors can be achieved.

Description

ANTITUMOR FORMULA AND EXTRACTION METHOD THEREOF

Technical Field

The present invention relates to the technical field of traditional Chinese medicine, and particularly relates to an antitumor formula and an extraction method thereof.

Background

With the increasingly severe environmental pollution of the earth, prevalence of tumors has been increased, and thus the life and living of people are seriously affected. Tumor treatment methods in modem medicine mainly include operation, radiotherapy, chemotherapy, radio frequency and intervention. Some antitumor drugs are produced in domestic manufacturers, but most of the drugs are fungus-like wall-broken powder or simple compositions. Tumors are mainly treated by methods such as the chemotherapy, radiotherapy and operation in the modem medicine. However, lots of clinical treatments show that, toxic and side effects of different degrees may be caused during chemotherapy and radiotherapy, and normal human cells will be greatly killed while killing the tumors, thereby causing emesis, weakness, diarrhea, leucopenia, tissue edema, mental numbness and tissue necrosis to the human body. In recent years, tumor treatment of the traditional Chinese medicine is called a special medication. The traditional Chinese medicine believes that blood stasis, phlegm-damp agglomeration, internal heat toxin and vital qi deficiency are pathogenesis of tumors. According to etiology, pathogenesis and occurrence and development rules of the tumors, anticancer treatment of the traditional Chinese medicine mainly includes four basic rules of strengthening and consolidating body resistance, clearing away heat and toxic materials, promoting blood circulation to remove blood stasis and resolving hard lump. Tumor treatment mechanisms in

Description

traditional Chinese medicine have major effects of directly inhibiting and killing tumor cells, inhibiting tumor angiogenesis, inhibiting telomerase activity, inducing cell differentiation, regulating cell signaling, resisting microtubules, enhancing immunity and reversing multidrug resistance. The tumor treatment of traditional Chinese medicine has a long history and is unique in curative effect. A vast amount of clinical and experimental researches have proved that, the traditional Chinese medicine may relieve tumor symptoms, improve survival quality of patients and lower the death rate. However, defects such as slow efficacy and unobvious effect are widespread in existing tumor treatment methods. There are still no experience and result that can be worth popularizing and using at present. Therefore, how to provide an antitumor formula that is low in price, wide in medicinal source, excellent in curative effect and low in toxicity is a problem that urgently needs to be solved by those skilled in the art.

Summary In view of this, the present invention provides an antitumor formula that is low in price, wide in medicinal source, excellent in curative effect and low in toxicity and an extraction method thereof. To achieve the above purpose, technical solutions of the present invention are as follows: the antitumor formula includes the following raw materials in parts by weight: 10-20 parts of nucleic acids, 10-20 parts of lectin, 10-20 parts of polysaccharides, 10-20 parts of ginsenosides, 5-10 parts of lipids and 5-10 parts of sterols. The present invention has beneficial effects as follows: the antitumor formula in the present invention is in conformity with the "assistant and guide" principle required in traditional Chinese medicine prescriptions and compositions. The antitumor formula is precise in composition and appropriate in compatibility and is

Description

in conformity with therapeutic principles of "weakening induration", "eliminating stagnation", "treating retention with purgation" and "tonifying deficiency" during tumor treatment of the traditional Chinese medicine. With the adoption of various raw materials in the present invention, an effect of effectively treating tumors can be achieved. Preferably, the nucleic acids, lectin, polysaccharides, lipids and sterols are derived from Agaricus blazei extract. Preferably, an extraction method of the Agaricus blazei extract includes the following steps: (1) crushing Agaricus blazei dried to a constant weight state; screening the crushed Agaricus blazei through a sieve of 80-150 meshes; adding a wall-breaking enzyme agent; performing enzymolysis on the mixture in an ultrasonic generator under a frequency of 22-24 KHz so as to obtain liquid; adding distilled water in an amount of 18-25 times that of the mass of the liquid; fully stirring the liquid after dissolving; extracting the solution with a complex enzyme in a thermostatic water bath at 44-54°C for 46-56 min; performing enzyme deactivation on the extracting solution in boiling water for 5-7 min; centrifuging the solution at 2500-3000 r/min for 5-10 min; and filtering the solution so as to obtain filter residues and an extracting solution; (2) performing enzymolysis on the extracting solution; concentrating the extracting solution to 1/2-1/3 of the original volume; regulating an isoelectric point with hydrochloric acid until the pH value is equal to 2; filtering the solution to obtain nucleic acid and supernatant; regulating the supernatant until the pH value is equal to 8; and filtering the solution so as to obtain lectin and liquor; (3) performing alcohol precipitation on the liquor obtained in the step (2); and filtering the solution so as to obtain polysaccharides;

Description

(4) adding an organic solvent into the filter residues obtained in the step (1); centrifuging the solution to take the supernatant; and removing the solvent so as to obtain lipids and sterols; (5) adding an organic solvent into the lipids and sterols and stirring; standing and layering the solution; washing the solution to be neutral so as to obtain a neutral organic layer; eluting the solution by a silica-gel chromatography column; collecting the eluant; concentrating and removing the solvent so as to obtain the sterols; and confirming that the rest precipitates are the lipids. Preferably, in the step (1), the wall-breaking enzyme agent is composed of cellulase and amylase according to a mass ratio of (1-2):(1-4); and a mass ratio of the wall-breaking enzyme agent to the Agaricus blazei is (0.5-2):100. The complex enzymes in the steps (1) and (2) are both composed of neutral protease and papain according to a mass ratio of (1-3):(2-5); and a mass ratio of the complex enzymes to the Agaricus blazei is (0.1-1):100. The organic solvent in the steps (4) and (5) is n-butyl alcohol or propyl alcohol; and the dosage of the organic solvent is 5-8 times that of the filter residues or precipitates. The above technical solutions have beneficial effects as follows: according to the extraction method in the present invention, the various ingredients in the Agaricus blazei extract can be continuously extracted. Preferably, an extraction method of ginseng extract includes the following steps: respectively crushing ginseng into powder of 80-100 meshes and performing enzymolysis; respectively adding the ginseng powder into ethanol in an amount of 8-10 times that of the powder for soaking; heating the solution to 40-50°C; maintaining the temperature for 2-3 h; filtering to obtain an extracting solution; concentrating the obtained extract; adding water into the extract; extracting the solution with saturated n-butanol for 3-4 times; adding isovolumetric acetone into

A

Description

the extracting solution for precipitating; and drying the precipitate to obtain the ginseng extract. Preferably, a specific operation of the enzymolysis is as follows: 0.5-1% of amylase in an amount of 5-7 times that of the powder is added into the powder for performing enzymolysis for 12-24 h. Preferably, a mass concentration of the ethanol is 40-50%; and soaking time is 12-24 h. The above technical solutions have beneficial effects as follows: precipitation of active ingredients is greatly increased; and yield of each extract is increased. The various raw materials in the present invention have effects as follows: The Agaricus blazei is neutral in nature, sweet in taste and attributive to heart, lung, liver and kidney meridians, and has effects of invigorating the brain, diminishing inflammation, tonifying the kidney, regulating blood sugar level, improving diabetes, lowering cholesterol, enhancing energy, improving arteriosclerosis and preventing and treating cardiovascular diseases. The Agaricus blazei extract: 1, has a function of enhancing human immunity: the Agaricus blazei extract enhances the autoimmune function of the human body by enhancing functions of a mononuclear phagocyte system and has effects of inhibiting cell division and regulating immune system response, thereby retarding and interfering with virus growth; 2, promotes hematopoietic functions of human bone marrow: the Agaricus blazei extract can enable a peripheral hemoglobin concentration and the total numbers of blood platelets and white blood cells to tend to be normal by improving inhibition of chemotherapy to the hematopoietic functions of the bone marrow, has an effect of inhibiting tumor cells, and may strengthen body resistance and improving health when taken for a long time; 3, promotes effects of chemotherapeutics such as cyclophosphamide and 5-Fu; 4, has an effect of inhibiting cell proliferation interfering with leukemia, wherein physiologically active polysaccharides in the extract are suitable for treatment of

Description

childhood leukemia; 5, has an effect of protecting liver and kidney and can be taken for a long time; according to the above effects, the Agaricus blazei attracts considerable attention of the health industry in Japan. Due to special dual-conditioning health care effects of activating immune response and strengthening and consolidating body resistance, the Agaricus blazei extract is widely applied to patients; and 6, the Agaricus blazei has anticancer biological functions. The ginsenoside has effects of preventing and resisting tumors. The present invention further provides a preparation method of the antitumor formula. The preparation method includes the following steps: (1) weighing various raw materials of an antitumor composition in parts by weight for later use; (2) uniformly stirring and mixing nucleic acids, lectin, polysaccharides, ginsenosides, lipids and sterols to obtain the antitumor product. Preferably, in the step (2), a stirring temperature is 40-50°C; and time is 30-40 min. The present invention further provides an application of the antitumor composition, wherein the antitumor product and pharmaceutically acceptable carriers are prepared into any of common oral preparations; and the oral preparations are capsules, granules, oral liquid, pills or tablets. Through the above technical solutions, compared with the prior art, the present invention discloses the antitumor composition as well as the preparation method and the application thereof. The antitumor composition is compounded from various extracts serving as raw materials. The raw materials are used together according to an appropriate ratio, are nutritive materials having different molecular structures, and have partially identical medicinal effects and respectively outstanding medicinal effects. The various ingredients are organically combined together according to an appropriate ratio, can complement and facilitate one

Description

another in the curative effects, achieve respective effects to be applied to tumor treatment to the utmost extent and can reduce corresponding cost.

Detailed Description

Technical solutions in the embodiments of the present invention are described clearly and fully below in combination with the embodiments of the present invention. Apparently, the described embodiments are merely part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those ordinary skilled in the art without contributing creative labor will belong to the protection scope of the present invention. A preparation method of Agaricus blazei extract in embodiments below includes the following steps: (1) Agaricus blazei dried to a constant weight state was crushed; the crushed Agaricus blazei was screened through a sieve of 1000 meshes; a wall-breaking enzyme agent was added; enzymolysis was performed on the mixture in an ultrasonic generator under a frequency of 22 KHz so as to obtain liquid; distilled water in an amount of 18 times that of the mass of the liquid was added; the liquid was fully stirred after dissolving; the solution was extracted with a complex enzyme in a thermostatic water bath at 44°C for 46 min; enzyme deactivation was performed on the extracting solution in boiling water for 5 min; the solution was centrifuged at 2500 r/min for 5 min; and the solution was filtered so as to obtain filter residues and an extracting solution, wherein a mass ratio of cellulase to amylase was 1:1; a mass ratio of the wall-breaking enzyme agent to the Agaricus blazei was 0.5:100; the complex enzyme was composed of neutral protease and papain according to a mass ratio of 1:2; and a mass ratio of the complex enzyme to the Agaricus blazei was 0.1:100;

Description

(2) enzymolysis was performed on the extracting solution with the complex enzyme; the extracting solution was concentrated to 1/2 of the original volume; then, an isoelectric point was regulated with hydrochloric acid until the pH value was equal to 2; the solution was filtered to obtain nucleic acid and supernatant; the supernatant was regulated until the pH value was equal to 8; and the solution was filtered so as to obtain lectin and liquor, wherein the complex enzyme was composed of neutral protease and papain according to a mass ratio of 1:2; and a mass ratio of the complex enzyme to the Agaricus blazei was 0.3:100; (3) 75% of ethanol in an amount of 6 times that of the volume of the liquor was added into the liquor obtained in the step (2); centrifugation was performed at 4000 r/min for 5 min; and the solution was filtered so as to obtain polysaccharides; (4) an acetone solution in an amount of 5 times that of the mass of the filter residues was added into the filter residues obtained in the step (1); the solution was centrifuged for 5 min at 4000 r/min to take the supernatant; and the solvent was removed so as to obtain lipid and sterol precipitates; (5) n-butyl alcohol in an amount of 8 times that of the mass of the lipids and sterols was added into the lipids and sterols for stirring; the solution was subjected to standing and layering; the solution was washed to be neutral so as to obtain a neutral organic layer; the solution was eluted by a silica-gel chromatography column; the eluant was collected; the solvent was concentrated and removed so as to obtain the sterols; and the rest precipitates were the lipids. An extraction method of the ginseng extract includes the following steps: ginseng was crushed into powder of 80 meshes; 0.5% of amylase in an amount of 6 times that of the mass of the powder was added into the powder for performing enzymolysis for 12 h; the powder was respectively added into ethanol in an amount of 8 times that of the powder for soaking; the solution was heated to °C; the temperature was maintained for 2 h; the solution was filtered to obtain an extracting solution; the obtained extract was concentrated; water was added into

Q

Description

the extract; the solution was extracted with saturated n-butanol for 3 times; isovolumetric acetone was added into the extracting solution for precipitating; and the precipitate was dried at 70°C for 20 min to obtain the ginseng extract. Embodiment 1 An antitumor formula includes the following raw materials in gram by weight: g of nucleic acid, 10 g of lectin, 10 g of polysaccharide, 10 g of ginsenoside, 5 g lipid and 5 g of sterol. A preparation method of the above antitumor formula includes the following steps: (1) various raw materials were weighed for later use; (2) nucleic acids, lectin, polysaccharides, ginsenosides, lipids and sterols were stirred at 40°C for 35 min to obtain the antitumor product. Embodiment 2 An antitumor formula includes the following raw materials in gram by weight: g of nucleic acid, 20 g of lectin, 20 g of polysaccharide, 20 g of ginsenoside, 10 g lipid and 10 g of sterol. A preparation method of the above antitumor formula includes the following steps: (1) various raw materials were weighed for later use; (2) nucleic acids, lectin, polysaccharides, ginsenosides, lipids and sterols were stirred at 40°C for 35 min to obtain the antitumor product. Embodiment 3 An antitumor formula includes the following raw materials in gram by weight: g of nucleic acid, 15 g of lectin, 15 g of polysaccharide, 15 g of ginsenoside, 8 g lipid and 8 g of sterol. A preparation method of the above antitumor formula includes the following steps: (1) various raw materials were weighed for later use;

Description

(2) nucleic acids, lectin, polysaccharides, ginsenosides, lipids and sterols were stirred at 40°C for 35 min to obtain the antitumor product. Embodiment 4 An antitumor formula includes the following raw materials in gram by weight: 12 g of nucleic acid, 12 g of lectin, 12 g of polysaccharide, 16 g of ginsenoside, 7 g lipid and 9 g of sterol. A preparation method of the above antitumor formula includes the following steps: (1) various raw materials were weighed for later use; (2) nucleic acids, lectin, polysaccharides, ginsenosides, lipids and sterols were stirred at 40°C for 35 min to obtain the antitumor product. Reference 1 An antitumor drug includes 15 g of Prunella vulgaris, 15 g of Coptis chinensis, 8 g of Glycyrrhiza, 7 g of cortex cinnamomi, 7 g of fructus evodiae, 7 g of rhizoma zingiberis, 8 g of rhizoma sparganii, 8 g of curcuma zedoaria, 15 g of Angelica sinensis and 15 g of ginseng. A preparation method includes the steps: the above raw materials were uniformly stirred and mixed to obtain the antitumor drug. Antitumor pharmacodynamic study on the composition in the present invention Influence on growth of S180 solid tumor 50 mice were taken and subjected to subcutaneous vaccination with S180 tumor fluid; the mice were randomly divided into 5 groups on the next day; each group included 10 mice; the 5 groups respectively included a blank control group, a positive control group (cyclophosphamide), a low-dose product group in embodiment 1 of the present invention, a medium-dose product group in embodiment 1 of the present invention, and a high-dose product group in embodiment 1 of the present invention; and the mice were all weighed. The drug

Description

was diluted with distilled water; the mice were subjected to intragastric administration according to 0.2 mL/10g of weight; the operations were conducted continuously for 10 days each time; the mice were weighed on the next day after drug withdrawal; the animals were killed; subcutaneous tumor blocks were peeled; the tumor blocks were weighed; a tumor inhibition rate was calculated; and data was as shown in Table 1: The tumor inhibition rate (%) was equal to (1-mean tumor weight in the treatment group/mean tumor weight in the blank control group) x%; Table 1: Data of the tumor inhibition effects on S180 subcutaneous tumor-bearing mice Administration dose Mean tumor weight Tumor inhibition rate (%)

(g/mL) (g)

Blank control group 1.89

Positive control group 0.20 0.96 49.2

Low-dose group 0.15 0.58 69.3

Medium-dose group 0.25 0.44 76.7

High-dose group 0.50 0.32 83.1

It can be seen from Table 1 that, the product in the present invention can achieve an effect of effectively inhibiting tumors. Toxicity experiment To ensure subacute toxicity conditions of the traditional Chinese medicine composition in the present invention, the present experiment was conducted on rats. SD rats were randomly divided into 5 groups; each group included 20 rats; the antitumor products prepared in embodiments 1-4 of the present invention were respectively crushed and then diluted with distilled water; the rats were drenched with the antitumor products per day at a dose of 0.2-0.8 mL/10g; and the rats were observed within 30 d. In the experimental period, the rats in each group were normal in appetite, behavior and excrement and good in development and had no obvious toxic

Description

symptoms; and hemogram and serum indexes of each group were all in a normal range. When continuously fed with the drug at a dose of 3 times that of the clinical dosage for 30 d, the rats had no obvious toxic or side effects, which indicates that the clinical dosage of the drug in the present invention is safe. Each embodiment in the description is described in a progressive way. The difference of each embodiment from each other is the focus of explanation. The same and similar parts among all of the embodiments can be referred to each other. For the device disclosed by the embodiments, because the device corresponds to a method disclosed by the embodiments, the device is simply described. Refer to the description of the method part for the related part. The above description of the disclosed embodiments enables those skilled in the art to realize or use the present invention. Many modifications made to these embodiments will be apparent to those skilled in the art. General principles defined herein can be realized in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to these embodiments shown herein, but will conform to the widest scope consistent with the principles and novel features disclosed herein.

1 )

Claims

1. An antitumor formula, comprising the following raw materials in parts by weight: 10-20 parts of nucleic acids, 10-20 parts of lectin, 10-20 parts of polysaccharides, 10-20 parts of ginsenosides, 5-10 parts of lipids and 5-10 parts of sterols.

2. The antitumor formula according to claim 1, wherein the nucleic acids, lectin, polysaccharides, lipids and sterols are derived from Agaricus blazei extract.

3. The antitumor formula according to claim 2, wherein an extraction method of the Agaricus blazei extract comprises the following steps: (1) crushing Agaricus blazei dried to a constant weight state; screening the crushed Agaricus blazei through a sieve of 80-150 meshes; adding a wall-breaking enzyme agent; performing enzymolysis on the mixture in an ultrasonic generator under a frequency of 22-24 KHz so as to obtain liquid; adding distilled water in an amount of 18-25 times that of the mass of the liquid; fully stirring the liquid after dissolving; extracting the solution with a complex enzyme in a thermostatic water bath at 44-54°C for 46-56 min; performing enzyme deactivation on the extracting solution in boiling water for 5-7 min; centrifuging the solution at 2500-3000 r/min for 5-10 min; and filtering the solution so as to obtain filter residues and an extracting solution; (2) performing enzymolysis on the extracting solution with a complex enzyme; concentrating the extracting solution to 1/2-1/3 of the original volume; then, regulating an isoelectric point with hydrochloric acid until the pH value is equal to 2; filtering the solution to obtain nucleic acid and supernatant; regulating the supernatant until the pH value is equal to 8; and filtering the solution so as to obtain lectin and liquor; (3) performing alcohol precipitation on the liquor obtained in the step (2); and filtering the solution so as to obtain polysaccharides;

Claims

(4) adding an organic solvent into the filter residues obtained in the step (1); centrifuging the solution to take the supernatant; and removing the solvent so as to obtain lipids and sterols; (5) adding an organic solvent into the lipids and sterols and stirring; standing and layering the solution; washing the solution to be neutral so as to obtain a neutral organic layer; eluting the solution by a silica-gel chromatography column; collecting the eluant; concentrating and removing the solvent so as to obtain the sterols; and confirming that the rest precipitates are the lipids.

4. The antitumor formula according to claim 3, wherein in the step (1), the wall-breaking enzyme agent is composed of cellulase and amylase according to a mass ratio of (1-2):(1-4); and a mass ratio of the wall-breaking enzyme agent to the Agaricus blazei is (0.5-2):100; the complex enzymes in the steps (1) and (2) are both composed of neutral protease and papain according to a mass ratio of (1-3):(2-5); and a mass ratio of the complex enzymes to the Agaricus blazei is (0.1-1):100.

5. The antitumor formula according to any one of claims 1-4, wherein an extraction method of ginseng extract comprises the following steps: respectively crushing ginseng into powder of 80-100 meshes and performing enzymolysis; respectively adding the ginseng powder into ethanol in an amount of 8-10 times that of the powder for soaking; heating the solution to 40-50°C; maintaining the temperature for 2-3 h; filtering to obtain an extracting solution; concentrating the obtained extract; adding water into the extract; extracting the solution with saturated n-butanol for 3-4 times; adding isovolumetric acetone into the extracting solution for precipitating; and drying the precipitate to obtain the ginseng extract.

6. The antitumor formula according to claim 5, wherein a specific operation of the enzymolysis is as follows: 0.5-1% of amylase in an amount of 5-7 times that of the powder is added into the powder for performing enzymolysis for 12-24 h.

Claims

7. The antitumor formula according to claim 5, wherein a mass concentration of the ethanol is 40-50%; and soaking time is 12-24 h.

8. A preparation method of the antitumor formula, comprising the following steps: (1) weighing various raw materials of the antitumor formula of any one of claims 1-7 in parts by weight for later use; (2) uniformly stirring and mixing nucleic acids, lectin, polysaccharides, ginsenosides, lipids and sterols to obtain the antitumor product.

9. The preparation method of the antitumor formula according to claim 8, wherein in the step (2), a stirring temperature is 40-50°C; and time is 30-40 min.

10. An application of an antitumor composition, comprising the antitumor formula of any one of claims 1-7, or the antitumor product prepared by the preparation method of claim 8 or 9 and pharmaceutically acceptable carriers are prepared into any of common oral preparations, wherein the oral preparations are capsules, granules, oral liquid, pills or tablets.