Preparation method of a traditional Chinese medicine composition and its application and product

Abstract

The present invention belongs to the field of traditional Chinese medicine technology, and in particular to a preparation method and application and product of a composition of Chinese medicine. Preparation method, characterized in that, including first adding water and auxiliary agent to Fructus Astragali, Fructus Chebulae, Rhubarb, and Rhizoma Curcumae, soaking, extracting, filtering, combining filtrate, obtaining extract and medicinal residues; auxiliary agent including mass ratio of 2‑4:1 sophoroside ester and hydrogenated lecithin; then medicinal residues and nine fragrant insects, cloves, and wood fragrance are mixed, water and enzyme are added for enzymolysis, enzyme is inactivated, filtered, and enzymolysis liquid is obtained; finally, Cordyceps sinensis is crushed, extract and enzymolysis liquid are added for mixing, concentrated, and dried. The present invention uses a special process specific enzyme to extract water-extracted medicinal residues and nine fragrant insects, cloves, and wood fragrance, and its extract is compatible with the remaining components of the present invention, and can better improve the immunity of the body, improve thymus coefficient and spleen index.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and specifically relates to a preparation method of a traditional Chinese medicine composition and its application and product. Background Art

[0002] In recent years, people have made some progress in the research and development of cancer treatment drugs. The emergence of a large number of drugs has improved the treatment effect, but it has also brought a series of toxic side effects such as bone marrow suppression. Therefore, it becomes particularly important to protect the body's resistance, enhance immunity, and improve treatment effects.

[0003] Due to the difficulty in treating malignant tumors, for centuries, scholars from various countries have sought ways to improve the efficacy of radiotherapy and chemotherapy by improving surgery. At the same time, looking for valuable clues from traditional medicine and increasing the number of treatment options has become an important way to conquer malignant tumors.

[0004] At present, malignant tumors have entered the stage of comprehensive treatment, with surgery, radiotherapy, chemotherapy, traditional Chinese medicine and other comprehensive treatments to achieve the purpose of increasing efficacy and reducing toxicity. The comprehensive research of traditional Chinese medicine and ethnic medicine preparations has always been one of the effective methods for the tumor prevention and treatment community to find a solution to the refractory nature of malignant tumors. It has been increasingly valued in improving the body's immunity, strengthening the body's body, and improving survival rates. Summary of the invention

[0005] In view of the deficiencies in the prior art, the present invention provides a composition for treating cardiovascular diseases.

[0006] In order to achieve the purpose of the present invention, the technical solution adopted is as follows:

[0007] A method for preparing a Chinese medicine composition comprises the following steps:

[0008] (1) adding water and an additive to asafoetida, terminalia chebula, rhubarb, and turmeric, soaking, extracting, filtering, and combining the filtrate to obtain an extract and a residue; the additive comprises sophoroside ester and hydrogenated lecithin in a mass ratio of 2-4:1;

[0009] (2) Mix the medicinal residue with the Chinese wormwood, cloves, and costusroot, add water and enzymes for enzymolysis, inactivate the enzymes, filter, and concentrate to obtain an enzymolysis solution;

[0010] (3) Crush the Cordyceps sinensis, add the extract and the enzymatic hydrolysate, mix, concentrate, and dry to obtain the product.

[0011] Preferably, the amount of water used in step (1) is 5-8 times the sum of the mass of asafoetida, terminalia chebula, rhubarb and turmeric, and the mass ratio of the auxiliary agent to water is 0.5-1:100.

[0012] Preferably, the extraction temperature in step (1) is 50-70°C, the number of extractions is 1-3 times, and the time of each extraction is 0.5-2h.

[0013] Preferably, the amount of water used in step (2) is 3-5 times the sum of the mass of the nine-scented insect, cloves and costusroot, and the enzymes used in the enzymatic hydrolysis are pectinase, papain and aminopeptidase.

[0014] Preferably, the mass ratio of the enzyme to water is 0.02-0.06:1; the mass ratio of the pectinase, papain and aminopeptidase is 1:0.05-0.1:0.02-0.05.

[0015] Preferably, the enzymatic hydrolysis temperature in step (2) is 40-55° C., the enzymatic hydrolysis time is 3-6 h, and the enzymatic hydrolysis pH is 5-7.

[0016] Preferably, the temperature for inactivating the enzyme in step (2) is 90-95° C., and the inactivation time is 5-10 min.

[0017] Preferably, the powder in step (3) is sieved through a 80-100 mesh screen.

[0018] The second object of the present invention is to provide the use of the traditional Chinese medicine composition prepared by the above preparation method in the preparation of immunity improving drugs or anti-tumor drugs.

[0019] The third object of the present invention is to provide a traditional Chinese medicine composition prepared by the above preparation method, which comprises, by weight, 1-10 parts of asafoetida, 5-15 parts of nine-scented worms, 1-10 parts of cloves, 5-10 parts of costusroot, 10-20 parts of rhubarb, 10-20 parts of turmeric, 1-2 parts of cordyceps and 10-15 parts of terminalia chebula.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] (1) The preparation method of the present invention has been explored and studied, and it has been found that adding auxiliary agents to extract individual medicinal materials can increase the extraction rate of active components and improve the efficacy.

[0022] (2) The present invention uses asafoetida, terminalia chebula, rhubarb and turmeric to extract water, and the residues are hydrolyzed with scutellaria baicalensis, cloves and costusroot, and the cordyceps is crushed. If necessary, it can also be crushed by ultrafine wall breaking. The extract prepared by the present invention is used to treat model mice, and it is confirmed that the prescription of the present invention has a significant inhibitory effect on the proliferation of liver cancer cells in model mice. Compared with the commonly used chemical drugs, it has less toxicity and no drug resistance.

[0023] (3) The present invention uses a special process and specific enzymes to extract the water-extracted medicinal residues and nine-scented insects, cloves, and costusroot, and the combination with the other components of the present invention can better improve the body's immunity, increase the thymus coefficient and spleen index. DETAILED DESCRIPTION

[0024] The present invention is further described below in conjunction with specific embodiments. The raw materials in the following examples are all commercially available conventional raw materials. The papain manufacturer is Nanning Pangbo Bioengineering Co., Ltd., with a model of 100,000 u / g; the aminopeptidase manufacturer is Dongheng Huadao, with a specification of 30,000 u / g; the pectinase manufacturer is Dongheng Huadao, with a specification of 30,000 u / g; the sophoroside supplier is Xibao Biotechnology (Shanghai) Co., Ltd.; the hydrogenated lecithin supplier is Henan Qinqiang Biotechnology Co., Ltd.

[0025] Example 1

[0026] The Chinese medicine composition comprises 5 parts of asafoetida, 8 parts of scutellaria baicalensis, 8 parts of cloves, 8 parts of costusroot, 15 parts of rhubarb, 15 parts of turmeric, 1 part of cordyceps sinensis and 10 parts of terminalia chebula. The above are weight parts.

[0027] The preparation method comprises the following steps:

[0028] (1) Add 3 times the amount of water to asafoetida, terminalia chebula, rhubarb and turmeric, and then add 0.8% of the water mass of auxiliary agents (sophoroside esters and hydrogenated lecithin in a mass ratio of 3:1), soak for 30 minutes, extract twice at 60°C, each extraction time is 1 hour, filter, and combine the filtrate to obtain the extract and the residue;

[0029] (2) Mix the medicinal residue with nine-scented worm, cloves and costus root, add 3 times the amount of water, and then add 0.06% of the water weight of enzyme (pectinase, papain and aminopeptidase in a mass ratio of 1:0.06:0.04), and enzymolyze for 5 hours at 45°C and pH 6. After the enzymolysis is completed, heat it to 90°C and keep it warm for 10 minutes to inactivate the enzyme, filter it, and obtain the enzymolysis solution;

[0030] (3) Crush the Cordyceps sinensis, pass it through a 100-mesh sieve, add the extract and the enzymatic hydrolysate, mix them, concentrate under reduced pressure at 60°C, and dry them.

[0031] Example 2

[0032] The Chinese medicine composition comprises 10 parts of asafoetida, 15 parts of scutellaria baicalensis, 10 parts of cloves, 10 parts of costusroot, 10 parts of rhubarb, 10 parts of turmeric, 2 parts of cordyceps sinensis and 15 parts of terminalia chebula. The above are weight parts.

[0033] The preparation method comprises the following steps:

[0034] (1) Add 3 times the amount of water to asafoetida, terminalia chebula, rhubarb and turmeric, and then add 1% of the water mass of auxiliary agents (sophoroside esters and hydrogenated lecithin in a mass ratio of 4:1), soak for 30 minutes, extract at 70°C for 1 hour, filter, and obtain the extract and the residue;

[0035] (2) Mix the medicinal residue with the Chinese worm, cloves, and costus root, add 3 times the amount of water, and then add 0.02% enzyme (a mixture of pectinase, papain, and aminopeptidase in a mass ratio of 1:0.1:0.05) in water, and perform enzymolysis at 55°C and pH 7 for 6 h. After the enzymolysis is completed, heat it to 95°C and keep it warm for 5 min to inactivate the enzyme, filter it, and obtain the enzymolysis solution;

[0036] (3) Crush the Cordyceps sinensis and pass it through an 80-mesh sieve; add the extract and the enzymatic hydrolysate, mix, concentrate under reduced pressure at 60°C, and dry to obtain the product.

[0037] Example 3

[0038] The Chinese medicine composition comprises 1 part of asafoetida, 5 parts of scutellaria baicalensis, 1 part of clove, 5 parts of costusroot, 10 parts of rhubarb, 10 parts of turmeric, 1 part of cordyceps sinensis and 10 parts of terminalia chebula. The above are weight parts.

[0039] The preparation method comprises the following steps:

[0040] (1) Add 3 times the amount of water to asafoetida, terminalia chebula, rhubarb and turmeric, and then add 0.5% of the water mass of auxiliary agents (sophoroside esters and hydrogenated lecithin in a mass ratio of 2:1), soak for 30 minutes, extract twice at 50°C, each extraction time is 1.5 hours, filter, and combine the filtrate to obtain the extract and the residue;

[0041] (2) Mix the medicinal residue with nine-scented worm, cloves and costus root, add 3 times the amount of water, and then add 0.05% enzyme (pectinase, papain and aminopeptidase in a mass ratio of 1:0.04:0.02) in water, and perform enzymolysis at 40°C and pH 5 for 3 h. After the enzymolysis is completed, heat it to 90°C and keep it warm for 10 min to inactivate the enzyme, filter it, and obtain the enzymolysis solution;

[0042] (3) Crush the Cordyceps sinensis; add the extract and the enzymatic hydrolysate, concentrate under reduced pressure at 60°C, and dry to obtain the product.

[0043] The following comparative examples 1-4 are individual prescriptions screened in the early stage of the experiment.

[0044] Comparative Example 1

[0045] The difference between this comparative example and Example 1 is that the auxiliary agent is sophoroside ester.

[0046] The raw materials of the Chinese medicine composition are 5 parts of asafoetida, 8 parts of nine-scented worm, 8 parts of clove, 8 parts of costusroot, 15 parts of rhubarb, 15 parts of turmeric, 1 part of cordyceps sinensis, and 10 parts of terminalia chebula. The above are weight parts.

[0047] The preparation method comprises the following steps:

[0048] (1) Add 3 times the amount of water to asafoetida, terminalia chebula, rhubarb and turmeric, then add 0.8% of the water mass of the auxiliary agent sophoroside ester, soak for 30 minutes, extract twice at 60°C, each extraction time is 1 hour, filter, and combine the filtrate to obtain the extract and the residue;

[0049] (2) Mix the medicinal residue with nine-scented worm, cloves and costus root, add 3 times the amount of water, and then add 0.06% of the water weight of enzyme (pectinase, papain and aminopeptidase in a mass ratio of 1:0.06:0.04), and enzymolyze for 5 hours at 45°C and pH 6. After the enzymolysis is completed, heat it to 90°C and keep it warm for 10 minutes to inactivate the enzyme, filter it, and obtain the enzymolysis solution;

[0050] (3) Crush the Cordyceps sinensis, pass it through a 100-mesh sieve, add the extract and the enzymatic hydrolysate, mix them, concentrate under reduced pressure at 60°C, and dry them.

[0051] Comparative Example 2

[0052] The difference between this comparative example and Example 1 is that the auxiliary agent is hydrogenated lecithin.

[0053] The raw materials of the Chinese medicine composition are 5 parts of asafoetida, 8 parts of nine-scented worm, 8 parts of clove, 8 parts of costusroot, 15 parts of rhubarb, 15 parts of turmeric, 1 part of cordyceps sinensis, and 10 parts of terminalia chebula. The above are weight parts.

[0054] The preparation method comprises the following steps:

[0055] (1) Add 3 times the amount of water to asafoetida, terminalia chebula, rhubarb and turmeric, and then add 0.8% of the water mass of hydrogenated lecithin as an auxiliary agent, soak for 30 minutes, extract twice at 60°C, each extraction time is 1 hour, filter, and combine the filtrate to obtain the extract and the residue;

[0056] (2) Mix the medicinal residue with nine-scented worm, cloves and costus root, add 3 times the amount of water, and then add 0.06% of the water weight of enzyme (pectinase, papain and aminopeptidase in a mass ratio of 1:0.06:0.04), and enzymolyze for 5 hours at 45°C and pH 6. After the enzymolysis is completed, heat it to 90°C and keep it warm for 10 minutes to inactivate the enzyme, filter it, and obtain the enzymolysis solution;

[0057] (3) Crush the Cordyceps sinensis, pass it through a 100-mesh sieve, add the extract and the enzymatic hydrolysate, mix them, concentrate under reduced pressure at 60°C, and dry them.

[0058] Comparative Example 3

[0059] The difference between this comparative example and Example 1 is that the enzymes used are pectinase and papain in a mass ratio of 1:0.1.

[0060] The raw materials of the Chinese medicine composition are 5 parts of asafoetida, 8 parts of nine-scented worm, 8 parts of clove, 8 parts of costusroot, 15 parts of rhubarb, 15 parts of turmeric, 1 part of cordyceps sinensis, and 10 parts of terminalia chebula. The above are weight parts.

[0061] The preparation method comprises the following steps:

[0062] (1) Add 3 times the amount of water to asafoetida, terminalia chebula, rhubarb and turmeric, and then add 0.8% of the water mass of auxiliary agents (sophoroside esters and hydrogenated lecithin in a mass ratio of 3:1), soak for 30 minutes, extract twice at 60°C, each extraction time is 1 hour, filter, and combine the filtrate to obtain the extract and the residue;

[0063] (2) Mix the medicinal residue with nine-scented worm, cloves, and costus root, add 3 times the amount of water, and then add 0.06% of the water weight of enzyme (a mixture of pectinase and papain in a mass ratio of 1:0.1), and perform enzymolysis at 45°C and pH 6 for 5 hours. After the enzymolysis is completed, heat it to 90°C and keep it warm for 10 minutes to inactivate the enzyme, filter it, and obtain the enzymolysis solution;

[0064] (3) Crush the Cordyceps sinensis, pass it through a 100-mesh sieve, add the extract and the enzymatic hydrolysate, mix them, concentrate under reduced pressure at 60°C, and dry them.

[0065] Comparative Example 4

[0066] The difference between this comparative example and Example 1 is that the preparation method is different.

[0067] The raw materials of the Chinese medicine composition are 5 parts of asafoetida, 8 parts of nine-scented worm, 8 parts of clove, 8 parts of costusroot, 15 parts of rhubarb, 15 parts of turmeric, 1 part of cordyceps sinensis, and 10 parts of terminalia chebula. The above are weight parts.

[0068] The preparation method comprises the following steps:

[0069] (1) Take the nine-scented insect, extract it with 75% ethanol, and take the supernatant for later use;

[0070] (2) Grind asafoetida, cloves, costusroot, half of turmeric, and half of rhubarb into fine powder. Grind cordyceps into fine powder. Mix the above powders thoroughly and set aside.

[0071] (3) Take the Terminalia chebula and the remaining turmeric and rhubarb, add water and boil, filter, concentrate, add ethanol to the ethanol concentration of 75%, and let it stand in the refrigerator at 4℃ for 24 hours. Take the supernatant and mix it with the supernatant of the nematode, recover the ethanol to obtain the extract, mix it evenly with the reserved medicinal powder, and dry it.

[0072] In vitro experiment 1: Detect the inhibitory effect of the drug of the present invention on the growth of cancer cells cultured in vitro.

[0073] Distilled water was added to the group 1 of Example 1 of the present invention to prepare a 5 g / ml drug solution; lung adenocarcinoma (GLC-82), gastric adenocarcinoma (SGC-7901), and liver cancer (BEL-7402) cell lines were taken, and the cell lines in logarithmic growth phase were prepared into 2×10 4 / ML cell suspension, inoculate 96-well plate, plant 0.1ML in each well, place in 37℃ CO2 incubator for 24 hours, then add 0.1ml of the drug in Example 1, add 0.1ml of water to the blank control, and then place in 37℃ CO2 incubator for 48 hours, add 20ul of MTT solution (5mg / ml) to each well, continue incubation for 4 hours and then terminate the culture, discard the culture medium, add dimethyl sulfoxide (DMSO0.1ML / well), mix the solution thoroughly, and measure the light absorption value (OD value) of each well on a 570nm wavelength enzyme-linked immunosorbent assay (WELLSAN MK3 made in the United States). The inhibition rate calculation formula is as follows:

[0074] Inhibition rate % = (1-OD value of experimental group / OD value of control group) × 100%.

[0075] Results: According to calculation, the average inhibition rate of Example 1 on the growth of lung adenocarcinoma (GLC-82) cells was 57.3%, the average inhibition rate on the growth of gastric adenocarcinoma (SGC-7901) cells was 49.1%, and the average inhibition rate on the growth of liver cancer (BEL-7402) cells was 86.5%.

[0076] Animal experiment 1. In vivo life extension experiment of anti-liver cancer (H22) ascites mice

[0077] Drug source: This drug was provided by Yunnan Mingyang Pharmaceutical Co., Ltd.

[0078] The positive control drug fluorouracil (5-Fu) injection was produced by Shanghai Xudong Haipu Pharmaceutical Co., Ltd.

[0079] Experimental animals: ICR mice were provided by the animal room of Yunnan Baiyao Group Company.

[0080] Tumor strain source: Mouse hepatocarcinoma (Ha) ascites type was introduced from Beijing Cancer Institute.

[0081] Experimental groups: 8×10 cells of hepatocellular carcinoma (Hz) ascites cell suspension were injected intraperitoneally under sterile conditions. 6 / 0.2ml / mouse in ICR mice, each batch of experiments has nine groups, namely, Examples 1-3 (3 groups), Comparative Examples 1-4 (4 groups), positive control drug fluorouracil group, and model group. The model group consisted of 3 mice, and the other groups consisted of 6 mice of the same sex, weighing 20g±1g, randomly divided into groups, weighed, and gavage-administered the drugs starting the next day after inoculation of cancer cells.

[0082] Dosage method: Examples 1-3 (3 groups) and Comparative Examples 1-4 (4 groups) were given the composition prepared in the corresponding group, once a day, 5 g / kg each time, for a course of 10 days. The model group was given the same dose of saline, and the positive group was given 0.05 g / kg. After the experiment, the life extension rate was calculated, life extension rate % = (average survival days of the drug group - average survival days of the control group) / average survival days of the control group × 100%.

[0083] The results are shown in Table 1.

[0084] Table 1

[0085]

[0086] Conclusion: Compared with the model group, the life extension rate of the mice in Examples 1-3 was 85-105%, which was close to that of the positive group. Comparative Examples 1-4 were prepared using other processes, and their life extension rates for liver cancer (Hz) ascites model mice were inferior to those in Example 1, indicating that the composition of the present invention has a good extension rate for liver cancer model mice.

[0087] Animal experiment 2: In vivo immune experiment on mice with ascites and liver cancer (H22)

[0088] Drug source: This drug was provided by Yunnan Mingyang Pharmaceutical Co., Ltd.

[0089] The positive control drug fluorouracil (5-Fu) injection was produced by Shanghai Xudong Haipu Pharmaceutical Co., Ltd.

[0090] Experimental animals: ICR mice were provided by the animal room of Yunnan Baiyao Group Company.

[0091] Tumor strain source: Mouse hepatocarcinoma (Ha) ascites type was introduced from Beijing Cancer Institute.

[0092] Experimental groups: 8×10 cells of hepatocellular carcinoma (Hz) ascites cell suspension were injected intraperitoneally under sterile conditions. 6 / 0.2ml / mouse in ICR mice, each batch of experiments has 10 groups, namely, Examples 1-3 (3 groups), Comparative Examples 1-4 (4 groups), positive control fluorouracil group, blank group, and model group. Each batch of experiments consists of 6 male ICR mice, weighing 20g±2g, randomly divided into groups, weighed, and except for the blank group, the rest of the groups were gavaged and administered the drug the next day after inoculation of cancer cells.

[0093] Administration method: The compositions prepared in the corresponding groups were administered to the mice of Examples 1-3 (3 groups) and Comparative Examples 1-4 (4 groups), once a day, 5 g / kg each time, for 10 consecutive days. The blank group and the model group were administered with the same dose of saline, and the positive group was administered with 0.05 g / kg. The mice were killed by orbital bleeding the next day after the last administration, and the thymus and spleen were removed and weighed to calculate the organ index (mg / 10 g body weight).

[0094]

[0095] Conclusion: The thymus index and spleen index of the model group mice were lower than those of the normal group, and p < 0.05. The thymus index of Examples 1-3 was close to that of the normal group, and the spleen index was higher than that of the normal group, which was significantly improved compared with the model group, indicating that the composition of the present invention can improve and enhance the body's immunity. Comparative Examples 1-4 are different from Example 1 in that they are prepared using other processes, and their efficacy in improving immunity is stronger.

[0096] The above detailed description is a specific description of one feasible embodiment of the present invention. The embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or modification that does not deviate from the present invention should be included in the scope of the technical solution of the present invention.

Claims

1. A method for preparing an anti-liver cancer Chinese medicine composition, characterized in that: The Chinese medicine composition is composed of the following raw materials by weight: 1-10 parts of asafoetida, 5-15 parts of scutellaria baicalensis, 1-10 parts of cloves, 5-10 parts of costusroot, 10-20 parts of rhubarb, 10-20 parts of turmeric, 1-2 parts of cordyceps sinensis and 10-15 parts of terminalia chebula; The preparation method comprises the following steps: (1) adding water and an auxiliary agent to asafoetida, terminalia chebula, rhubarb, and turmeric, soaking, extracting, filtering, and combining the filtrate to obtain an extract and a residue; the auxiliary agent is a sophoroside ester and hydrogenated lecithin in a mass ratio of 2-4:1; (2) Mix the medicinal residue with the Chinese wormwood, cloves, and costusroot, add water and enzymes for enzymolysis, inactivate the enzymes, filter, and obtain an enzymolysis solution; (3) Crush the Cordyceps sinensis, add the extract and the enzymatic solution, mix, concentrate, and dry to obtain; In step (1), the extraction temperature is 50-70°C, the number of extractions is 1-3 times, and the time of each extraction is 0.5-2h; In step (1), the mass ratio of the auxiliary agent to water is 0.5-1:100; In step (2), the enzymes used for enzymolysis are pectinase, papain and aminopeptidase; the mass ratio of the enzyme to water is 0.02-0.06:100; the mass ratio of the pectinase, papain and aminopeptidase is 1:0.05-0.1:0.02-0.05; The enzymatic hydrolysis temperature in step (2) is 40-55°C, the enzymatic hydrolysis time is 3-6h, and the enzymatic hydrolysis pH is 5-7.

2. The preparation method according to claim 1, characterized in that: The amount of water used in step (1) is 3-8 times the sum of the mass of asafoetida, terminalia chebula, rhubarb and turmeric.

3. The preparation method according to claim 1, characterized in that: The amount of water used in step (2) is 3-5 times the sum of the mass of the nine-scented insect, cloves and costusroot.

4. The preparation method according to claim 1, characterized in that: The temperature for inactivating the enzyme in step (2) is 90-95° C., and the inactivation time is 5-10 min.

5. The preparation method according to claim 1, characterized in that: After the pulverization in step (3), the product is sieved through a 80-100 mesh sieve.

6. Use of a traditional Chinese medicine composition prepared by the preparation method according to any one of claims 1 to 5 in the preparation of anti-liver cancer drugs.

7. An anti-liver cancer Chinese medicine composition prepared by the preparation method according to any one of claims 1 to 5.

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