Traditional Chinese medicine composition as well as preparation method and application thereof

Abstract

The invention provides a traditional Chinese medicine composition as well as a preparation method and application thereof. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 15-30 parts of herba artemisiae scopariae; 7.5 to 15 parts of radix astragali; 7.5-15 parts of glossy privet fruit; 7.5-15 parts of semen plantaginis; 5.0 to 10 parts of madder; 7.5 to 15 parts of Chinese yam; 5.0 to 10 parts of chrysanthemum; 5.0 to 10 parts of hawthorn; 5.0 to 10 parts of medulla stachyuri; 7.5 to 15 parts of polyporus umbellatus; and 5.0-10 parts of cortex phellodendri. The traditional Chinese medicine composition adopts the specific components and proportions, has a synergistic effect, and has the effects of clearing away heat and toxic materials, promoting diuresis, relieving jaundice, soothing the liver and tonifying qi. Hepatitis disease signal channels can be improved, HepG2.215 cell hepatitis B surface antigen and hepatitis B core antigen secretion can be reduced, and hepatitis virus replication can be potentially inhibited. The compound is used for treatment and adjuvant treatment of viral hepatitis, especially hepatitis B or hepatitis E virus infection.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a traditional Chinese medicine composition, a preparation method and application thereof. Background Art

[0002] Viral hepatitis is an infectious disease caused by a variety of hepatitis viruses and characterized by liver lesions. Clinically, the main manifestations are loss of appetite, nausea, upper abdominal discomfort, pain in the liver area, and fatigue. Some patients may have jaundice, fever, and hepatomegaly with liver damage. Some patients may develop chronic disease and even cirrhosis, and a few may develop liver cancer. There are about 2 million new cases of hepatitis in my country each year. Among them, hepatitis A accounts for 50%, hepatitis B accounts for 20%, hepatitis C accounts for 5%-10%, and hepatitis E accounts for 10%. Chronic hepatitis B, often referred to as "hepatitis B", is caused by infection with hepatitis B virus (HBV). If the infection persists for more than 6 months, the liver will show varying degrees of inflammation, necrosis, and / or liver fibrosis. There are about 257 million chronic HBV infections worldwide, and about 887,000 people die from HBV infection each year. The HBsAg prevalence rate in China is 5% to 6%, and there are about 70 million chronic HBV infections, of which about 20 million to 30 million are chronic hepatitis B patients. Anti-HBV treatment is the key to preventing further development of liver damage. At present, anti-HBV drugs are mainly interferon and antiviral drugs. These drugs have rapid efficacy, but are prone to relapse and have certain side effects. There is no specific drug for the prevention of liver damage and the relief of liver inflammation.

[0003] Hepatitis E is an infectious disease with hepatitis E virus (HEV) as the pathogen, which is transmitted among the population through the fecal-oral route. Although most cases of HEV infection clinically manifest as self-limiting acute infections and have a low mortality rate in young people (approximately 0.5%-3%), the mortality rate of pregnant women infected with HEV is close to 30%, and some patients (especially those with immunosuppression) will develop chronic diseases. Large-scale outbreaks and epidemics of hepatitis E usually occur in developing countries and areas with poor sanitary conditions, and in developed countries, they are usually sporadic cases. According to statistics from the World Health Organization, more than 20 million people are infected with HEV each year, of which more than 3 million will show symptoms of acute hepatitis and cause about 56,600 deaths.

[0004] There is no specific drug that can completely cure hepatitis B in the world. There is no specific drug for hepatitis E infection, and clinical treatment is mainly symptomatic. For viral hepatitis, the current clinical medication is mainly to inhibit viral replication, relieve symptoms, and improve liver function, mainly including antiviral drugs, non-specific liver protection drugs, enzyme-lowering drugs, etc., among which antiviral drugs are the most important. At present, there are nearly 50 antiviral drugs developed at home and abroad, but only a few nucleoside drugs such as acyclovir and lamivudine are actually used in clinical practice. This type of drug can usually only delay viral replication, and has serious defects such as large toxic side effects, high prices, and drug rebound after discontinuation; and liver protection and enzyme-lowering drugs usually have no obvious effect in inhibiting viral replication.

[0005] Therefore, it is very necessary to provide a drug that can be used to treat hepatitis virus, especially hepatitis B virus or hepatitis E virus infection and a preparation method thereof. Summary of the invention

[0006] In view of this, the Chinese medicine composition provided by the present invention has the effect of synergistically inhibiting the replication of hepatitis viruses and improving the abnormal signal pathways after hepatitis E virus infects liver cells, and is used for the treatment and adjuvant treatment of hepatitis B or hepatitis E.

[0007] The present invention provides a traditional Chinese medicine composition, comprising the following raw materials in parts by weight:

[0008] Artemisia capillaris 15-30; Astragalus 7.5-15; Ligustrum lucidum 7.5-15; Plantago asiatica 7.5-15; Rubia cordifolia 5.0-10; Dioscorea opposita 7.5-15; Chrysanthemum 5.0-10; Crataegus pinnatifida 5.0-10; Smilax glabra 5.0-10; Poria 7.5-15; Phellodendron chinense 5.0-10.

[0009] Preferably, the Chinese medicine composition comprises the following raw materials in parts by weight:

[0010] Artemisia capillaris 16-29; Astragalus 8-14; Ligustrum lucidum 8-14; Plantago asiatica 8-14; Rubia cordifolia 5.0-9; Dioscorea opposita 8-14; Chrysanthemum 5.0-9; Crataegus pinnatifida 5.0-9; Smilax glabra 5.0-9; Poria 8-14; Phellodendron chinense 5.0-9.

[0011] Preferably, the Chinese medicine composition comprises the following raw materials in parts by weight:

[0012] Artemisia capillaris 17-28; Astragalus 9-13; Ligustrum lucidum 9-12; Plantago asiatica 9-13; Rubia cordifolia 6-9; Dioscorea opposita 8-13; Chrysanthemum 6-9; Crataegus pinnatifida 6-9; Smilax glabra 6-9; Poria 9-13; Phellodendron chinense 6-9.

[0013] The present invention provides a method for preparing a Chinese medicine composition according to any one of the above technical solutions, comprising:

[0014] Mix Artemisia capillaris, Herba Artemisiae Scopariae, Polyporus, Seed of Plantago, Cortex Phellodendri, Flos Chrysanthemi, Fructus Hawthorne, Rubia cordifolia, Astragalus, Fructus Ligustri Lucidi, Dioscorea opposita and water, heat to extract, filter, concentrate and dry to obtain the product.

[0015] Preferably, the mass ratio of the total amount of Artemisia capillaris, Herba Artemisiae Scopariae, Polyporus, Semen Plantaginis, Cortex Phellodendri, Flos Chrysanthemi, Fructus Crataegi, Rubia cordifolia, Astragalus, Fructus Ligustri Lucidi and Dioscorea opposita to water is 1:(10-15).

[0016] Preferably, the temperature of the heating extraction is 100° C., the time is 1 to 1.5 hours, and the number of times of the heating extraction is 1 to 3 times.

[0017] Preferably, the concentration is concentrated at 60-70° C. to 1.1-1.2 g / mL;

[0018] The drying is specifically: spray drying at an inlet air temperature of 130°C;

[0019] The concentration and drying process further includes crushing, wherein the crushing value is 50 to 300 meshes.

[0020] The present invention provides use of any of the above-mentioned compositions or a composition prepared by any of the above-mentioned technical solutions in preparing a drug for treating viral hepatitis.

[0021] Preferably, the treatment includes improving abnormal signaling pathways after hepatitis E virus infects liver cells, reducing the secretion of hepatitis B surface antigen and hepatitis B core antigen in HepG2.215 cells, and potentially inhibiting hepatitis virus replication.

[0022] The present invention provides a product that can be used for viral hepatitis, comprising the composition described in any one of the above technical solutions or a composition prepared by the preparation method described in any one of the above technical solutions.

[0023] Preferably, the pharmaceutical dosage form includes, but is not limited to, one or more of granules, decoctions, powders, oral solutions, tablets, pills, pastes or capsules;

[0024] Compared with the prior art, the present invention provides a Chinese medicine composition, comprising the following raw materials in parts by weight: 15-30 parts of Artemisia capillaris; 7.5-15 parts of Astragalus; 7.5-15 parts of Ligustrum lucidum; 7.5-15 parts of Plantago; 5.0-10 parts of Rubia cordifolia; 7.5-15 parts of Dioscorea opposita; 5.0-10 parts of Chrysanthemum; 5.0-10 parts of Crataegus pinnatifida; 5.0-10 parts of Herba Glehniae; 7.5-15 parts of Polyporus; 5.0-10 parts of Cortex Phellodendri. The present invention adopts the above-mentioned specific components and proportions, has a synergistic effect, can improve the abnormal signal pathway after hepatitis E virus infects liver cells, reduce the secretion of hepatitis B surface antigen and hepatitis B core antigen in HepG2.215 cells, and potentially inhibit the replication of hepatitis virus, and is used for the treatment and adjuvant treatment of hepatitis B or hepatitis E. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 . The Chinese medicines in the prescription of the present invention improve the abnormal disease signal pathways after hepatitis E virus infects liver cells;

[0026] Figure 2 . The invention and the comparative prescription improve 24 abnormal disease signal pathways after hepatitis E virus infects liver cells;

[0027] Figure 3 .3 The results of the in vitro anti-cytotoxicity and hepatitis B virus replication tests of the first prescription;

[0028] Figure 4 . Test results of cytotoxicity and anti-HBV replication of different compound prescriptions;

[0029] Figure 5 . Different prescription compositions improve 24 abnormal disease signal pathways after hepatitis E virus infects liver cells. DETAILED DESCRIPTION

[0030] The present invention provides a Chinese medicine composition, a preparation method and application thereof, and those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar replacements and modifications are obvious to those skilled in the art, and they all fall within the scope of protection of the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the method and application of this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0031] The present invention provides a traditional Chinese medicine composition, comprising the following raw materials in parts by weight:

[0032] Artemisia capillaris 15-30; Astragalus 7.5-15; Ligustrum lucidum 7.5-15; Plantago asiatica 7.5-15; Rubia cordifolia 5.0-10; Dioscorea opposita 7.5-15; Chrysanthemum 5.0-10; Crataegus pinnatifida 5.0-10; Smilax glabra 5.0-10; Poria 7.5-15; Phellodendron chinense 5.0-10.

[0033] The present invention does not limit the source of the above-mentioned traditional Chinese medicine, and any commercially available one may be used.

[0034] The raw materials of the traditional Chinese medicine composition provided by the present invention include 15 to 30 parts by weight of Artemisia capillaris, preferably 16 to 29 parts by weight, and more preferably 17 to 28 parts by weight.

[0035] Artemisia capillaris can clear away damp-heat, promote bile secretion and reduce jaundice. It is used for jaundice, oliguria, damp-heat, summer dampness, and itching caused by eczema.

[0036] The raw materials of the traditional Chinese medicine composition provided by the present invention include 7.5 to 15 parts by weight of astragalus, preferably 8 to 14 parts by weight, and more preferably 9 to 13 parts by weight.

[0037] Astragalus can replenish Qi and raise Yang, consolidate the exterior and stop sweating, promote diuresis and reduce swelling, promote the production of body fluids and nourish blood, relieve stagnation and numbness, expel toxins and discharge pus, and heal sores and promote tissue regeneration. It is used for Qi deficiency and fatigue, poor appetite and loose stools, sinking of Qi in the middle, long-term diarrhea and prolapse of the anus, bloody stool and metrorrhagia, spontaneous sweating due to exterior deficiency, Qi deficiency and edema, internal heat and thirst, blood deficiency and sallow complexion, hemiplegia, pain and numbness, and carbuncles that are difficult to heal and that do not heal for a long time.

[0038] The raw materials of the traditional Chinese medicine composition provided by the invention include 7.5 to 15 parts by weight of Ligustrum lucidum fruit, preferably 8 to 14 parts by weight, and more preferably 9 to 12 parts by weight.

[0039] Fructus Ligustri Lucidi nourishes the liver and kidneys, improves eyesight and darkens hair. It is used for liver and kidney yin deficiency, dizziness and tinnitus, soreness of waist and knees, premature graying of hair, blurred vision, internal heat and thirst, and bone steaming and hot flashes.

[0040] The raw materials of the traditional Chinese medicine composition provided by the present invention include 7.5 to 15 parts by weight of Plantago seeds; preferably include 8 to 14 parts by weight; more preferably include 9 to 13 parts by weight.

[0041] Plantago seed, clears away heat, promotes diuresis and relieves stranguria, eliminates dampness and stops diarrhea, improves eyesight and eliminates phlegm. It is used for painful stranguria caused by heat, edema, diarrhea caused by summer dampness, red and swollen eyes, and cough caused by phlegm and heat.

[0042] The raw materials of the traditional Chinese medicine composition provided by the invention include 5.0 to 10 parts by weight of madder, preferably 5 to 9 parts by weight, and more preferably 6 to 9 parts by weight.

[0043] Rubia cordifolia, cools blood, removes blood stasis, stops bleeding and promotes menstruation. It is used for hematemesis, epistaxis, metrorrhagia, traumatic bleeding, blood stasis, amenorrhea, joint pain, and swelling and pain from falls.

[0044] The raw materials of the traditional Chinese medicine composition provided by the present invention include 7.5 to 15 parts by weight of yam, preferably 8 to 14 parts by weight, and more preferably 8 to 13 parts by weight.

[0045] Chinese yam nourishes the spleen and stomach, promotes the production of body fluids and benefits the lungs, nourishes the kidneys and astringes semen, and is used for spleen deficiency, poor appetite, chronic diarrhea, asthma and cough due to lung deficiency, spermatorrhea due to kidney deficiency, leucorrhea, frequent urination, and thirst due to deficiency heat. Chinese yam stir-fried with bran nourishes the spleen and stomach, and is used for spleen deficiency, poor appetite, diarrhea, loose stools, and excessive leucorrhea.

[0046] The raw materials of the traditional Chinese medicine composition provided by the present invention include 5.0 to 10 parts by weight of chrysanthemum, preferably 5 to 9 parts by weight, and more preferably 6 to 9 parts by weight.

[0047] Chrysanthemum can dispel wind and heat, soothe the liver and improve eyesight, clear away heat and detoxify. It is used for wind-heat colds, headaches and dizziness, red and swollen eyes, blurred vision, sores, carbuncles and swelling.

[0048] The raw materials of the traditional Chinese medicine composition provided by the present invention include 5.0 to 10 parts by weight of hawthorn, preferably 5 to 9 parts by weight, and more preferably 6 to 9 parts by weight.

[0049] Hawthorn can help digestion and invigorate the stomach, promote qi and dissipate blood stasis, remove turbidity and reduce fat. It is used for meat stagnation, stomach distension, diarrhea and abdominal pain, blood stasis and amenorrhea, postpartum blood stasis, heart and abdomen stabbing pain, chest pain, hernia pain, and hyperlipidemia. Charred hawthorn has a stronger effect on digestion and stagnation, and is used for meat stagnation and diarrhea.

[0050] The raw materials of the traditional Chinese medicine composition provided by the invention include 5.0 to 10 parts by weight of cyperus rotundus, preferably 5 to 9 parts by weight, and more preferably 6 to 9 parts by weight.

[0051] Xiaotongcao, clears away heat, promotes diuresis and promotes lactation. It is used for urinary difficulty, stranguria and lack of milk.

[0052] The raw materials of the traditional Chinese medicine composition provided by the present invention include 7.5 to 15 parts by weight of Polyporus umbellatus, preferably 8 to 14 parts by weight, and more preferably 9 to 13 parts by weight.

[0053] Poria, a diuretic and dampness-eliminating herb, is used for dysuria, edema, diarrhea, stranguria, and leucorrhea.

[0054] The raw materials of the traditional Chinese medicine composition provided by the present invention include 5.0-10 parts by weight of Phellodendron chinense, preferably 5-9 parts by weight, and more preferably 6-9 parts by weight.

[0055] Huangbai, clears away heat and dampness, purges fire and eliminates steam, detoxifies and treats sores. It is used for damp-heat diarrhea, jaundice and red urine, leucorrhea and itching, hot and painful stranguria, beriberi and paralysis, bone steaming and fatigue fever, night sweats and spermatorrhea, sores, swelling and poison, eczema and wet sores. Salted Huangbai nourishes yin and reduces fire, and is used for yin deficiency and excessive fire, night sweats and bone steaming.

[0056] The present invention improves the hepatitis disease signal pathway, reduces the secretion of hepatitis B surface antigen and hepatitis B core antigen of HepG2.215 cells through the synergistic effect of the above-mentioned components, potentially inhibits the replication of hepatitis viruses, and is used for the treatment and adjuvant treatment of hepatitis B or hepatitis E. Among them, this prescription has the effects of clearing away heat and detoxifying, removing dampness and jaundice, and soothing the liver and replenishing qi. Among them, Artemisia capillaris clears dampness and heat, promotes gallbladder and relieves jaundice as the main medicine, small tongcao, plantago seed, and umbellate promote water and infiltrate dampness, and Phellodendron chinense and chrysanthemum clear heat and detoxify, and suppress liver yang, which are all ministerial medicines. Astragalus, yam, and Ligustrum lucidum replenish qi and nourish yin, reduce swelling and relieve pain, and Rubia cordifolia and hawthorn soothe the liver, promote qi, and disperse blood stasis as adjuvants.

[0057] In some preferred embodiments of the present invention, the Chinese medicine composition comprises the following raw materials in parts by weight:

[0058] Artemisia capillaris 16-29; Astragalus 8-14; Ligustrum lucidum 8-14; Plantago asiatica 8-14; Rubia cordifolia 5.0-9; Dioscorea opposita 8-14; Chrysanthemum 5.0-9; Crataegus pinnatifida 5.0-9; Smilax glabra 5.0-9; Poria 8-14; Phellodendron chinense 5.0-9.

[0059] In some preferred embodiments of the present invention, the Chinese medicine composition comprises the following raw materials in parts by weight:

[0060] Artemisia capillaris 17-28; Astragalus 9-13; Ligustrum lucidum 9-12; Plantago asiatica 9-13; Rubia cordifolia 6-9; Dioscorea opposita 8-13; Chrysanthemum 6-9; Crataegus pinnatifida 6-9; Smilax glabra 6-9; Poria 9-13; Phellodendron chinense 6-9.

[0061] In some preferred embodiments of the present invention, the Chinese medicine composition comprises the following raw materials in parts by weight:

[0062] Artemisia capillaris 30; Astragalus 15; Ligustrum lucidum 15; Plantago asiatica 15; Rubia cordifolia 10; Dioscorea opposita 15; Chrysanthemum 10; Crataegus pinnatifida 10; Smilax glabra 10; Poria 15; Phellodendron chinense 10.

[0063] In some preferred embodiments of the present invention, the Chinese medicine composition comprises the following raw materials in parts by weight:

[0064] Artemisia capillaris 15; Astragalus 7.5; Ligustrum lucidum 7.5; Plantago asiatica 7.5; Rubia cordifolia 7.5; Dioscorea opposita 7.5; Chrysanthemum 7.5; Crataegus pinnatifida 7.5; Glehnia littoralis 7.5; Poria 7.5; Phellodendron chinense 7.5.

[0065] In some preferred embodiments of the present invention, the Chinese medicine composition comprises the following raw materials in parts by weight:

[0066] Artemisia capillaris 22.5; Astragalus 10; Ligustrum lucidum 10; Plantago asiatica 10; Rubia cordifolia 5; Dioscorea opposita 12; Chrysanthemum 5; Crataegus pinnatifida 5; Smilax glabra 5; Poria 12; Phellodendron chinense 5.

[0067] The present invention creatively combines the above 11 traditional Chinese medicines in a reasonable manner, so that the components play a synergistic role, which can improve the hepatitis disease signaling pathway, reduce the secretion of hepatitis B surface antigen and hepatitis B core antigen in HepG2.215 cells, and potentially inhibit the replication of hepatitis viruses.

[0068] The present invention provides a method for preparing a Chinese medicine composition according to any one of the above technical solutions, comprising:

[0069] Mix Artemisia capillaris, Herba Artemisiae Scopariae, Polyporus, Seed of Plantago, Cortex Phellodendri, Flos Chrysanthemi, Fructus Hawthorne, Rubia cordifolia, Astragalus, Fructus Ligustri Lucidi, Dioscorea opposita and water, heat to extract, filter, concentrate and dry to obtain the product.

[0070] The above components and proportions provided by the present invention have been clearly described and will not be repeated here.

[0071] The mass ratio of the total amount of Artemisia capillaris, Herba Artemisiae Sinensis, Polyporus, Plantago, Phellodendron, Chrysanthemum, Crataegus, Rubia cordifolia, Astragalus, Fructus Ligustri Lucidi and Dioscorea opposita to water is 1:(10-15); specifically, it can be 1:10, 1:11, 1:12, 1:13, 1:14 or 1:15; or a point value between any two of the above.

[0072] The temperature of the heating extraction is 100° C., the time is 1 to 1.5 hours, and the number of times of the heating extraction is 1 to 3 times.

[0073] After heating extraction, the drug solutions are combined.

[0074] Preferably, the powders are weighed according to weight and mixed thoroughly, placed in a round-bottom flask equipped with an electric heating jacket, 10 to 15 times the volume of ultrapure water is added and stirred, heated to boiling, and then the temperature is adjusted to maintain a slightly boiling state and continue heating for 1 to 1.5 hours, filtered through 200-mesh gauze to obtain extract 1 and stored for later use. Repeat the above steps to obtain extract 2 and store it for later use. Combine extracts 1 and 2.

[0075] The combined medicinal solutions are concentrated and dried to obtain a composition extract.

[0076] Preferably, the method can be as follows: after concentrating under reduced pressure using a rotary evaporator to a density of 1.1-1.2 g / mL (60-70°C), spray drying is performed at an inlet air temperature of 130°C, and then the spray-dried powder is crushed to obtain a 50-300 mesh powder, which is then packed into an aluminum foil bag, sealed and labeled, and stored in a moisture-proof cabinet.

[0077] The present invention provides use of any of the above-mentioned compositions or a composition prepared by any of the above-mentioned technical solutions in preparing a drug for treating viral hepatitis.

[0078] The treatment includes improving hepatitis disease signaling pathways, reducing the secretion of hepatitis B surface antigen and hepatitis B core antigen in HepG2.215 cells, and potentially inhibiting hepatitis virus replication.

[0079] The above cells include HepG2.

[0080] The present invention can be used for the treatment and auxiliary treatment of hepatitis virus, especially HBV and HEV infection.

[0081] The present invention provides a product for treating viral hepatitis, comprising the composition described in any one of the above technical solutions or a composition prepared by the preparation method described in any one of the above technical solutions.

[0082] That is, the present invention provides a medicine for treating liver diseases, including the composition described in any one of the above technical solutions or the composition prepared by any one of the above preparation methods.

[0083] The drug of the present invention has the functions of improving the signal pathway of hepatitis disease, reducing the secretion of hepatitis B surface antigen and hepatitis B core antigen of HepG2.215 cells, and potentially inhibiting the replication of hepatitis virus.

[0084] The signal pathways used in the present invention are specifically complement and coagulation cascade reactions, fat digestion and absorption, arginine biosynthesis, cholesterol metabolism, oxidative phosphorylation, PPAR signaling pathway, propionate metabolism, carbon metabolism, cytochrome P450 metabolism of exogenous substances, non-alcoholic fatty liver disease, fatty acid degradation, drug metabolism-cytochrome P450, valine, leucine and isoleucine degradation, amino acid biosynthesis, citric acid cycle (TCA cycle), bile secretion, pyruvate metabolism, adipose cytokine signaling pathway, hepatitis C, apoptosis, hepatitis B, shigellosis, Salmonella infection, and eukaryotic ribosome biosynthesis.

[0085] The present invention also includes auxiliary materials or ingredients commonly used in the pharmaceutical field.

[0086] The dosage form of the present invention includes, but is not limited to, one or more of granules, decoctions, powders, oral liquids, tablets, pills, pastes or capsules.

[0087] The present invention provides a Chinese medicine composition, comprising the following raw materials in parts by weight: 15-30 parts of Artemisia capillaris; 7.5-15 parts of Astragalus; 7.5-15 parts of Ligustrum lucidum; 7.5-15 parts of Plantago; 5.0-10 parts of Rubia cordifolia; 7.5-15 parts of Dioscorea opposita; 5.0-10 parts of Chrysanthemum; 5.0-10 parts of Crataegus pinnatifida; 5.0-10 parts of Herba Glehniae; 7.5-15 parts of Polyporus; and 5.0-10 parts of Cortex Phellodendri. The present invention adopts the above-mentioned specific components and proportions, has a synergistic effect, can activate the hepatitis disease signaling pathway, reduce the secretion of hepatitis B surface antigen and hepatitis B core antigen of HepG2.215 cells, and potentially inhibit the replication of hepatitis viruses, and is used for the treatment and adjuvant treatment of hepatitis B or hepatitis E.

[0088] In order to further illustrate the present invention, a Chinese medicine composition, a preparation method and application thereof provided by the present invention are described in detail below in conjunction with examples.

[0089] Example 1: Improvement of disease signaling pathways in HEV patients by prescription

[0090] (1) Abnormal signaling pathways after hepatitis E virus infection of liver cells

[0091] The transcriptome sequencing data (GSE135619) of human primary hepatocytes infected by HEV were retrieved from the NCBI (https: / / www.ncbi.nlm.nih.gov / ) database, and the sequencing data were subjected to KEGG pathway gene set enrichment analysis (GSEA), and 24 significantly enriched abnormal signal pathways closely related to HEV infection were selected. All or part of the significantly enriched signal pathways can be selected as abnormal signal pathways after hepatitis E virus infection of hepatocytes, that is, disease signal pathways. The signal pathways used in this patent are specifically complement and coagulation cascades, fat digestion and absorption, arginine biosynthesis, cholesterol metabolism, oxidative phosphorylation, PPAR signaling pathway, propionate metabolism, carbon metabolism, cytochrome P450 metabolism of exogenous substances, non-alcoholic fatty liver disease, fatty acid degradation, drug metabolism-cytochrome P450, valine, leucine and isoleucine degradation, amino acid biosynthesis, citric acid cycle (TCA cycle), bile secretion, pyruvate metabolism, adipose cytokine signaling pathway, hepatitis C, apoptosis, hepatitis B, shigellosis, Salmonella infection, and eukaryotic ribosome biosynthesis.

[0092] (2) Transcriptome sequencing of HepG2 cells treated with traditional Chinese medicine

[0093] The single medicine involved in the prescription of the present invention includes Artemisia capillaris, Herba Lycopodii, Polyporus, Plantago, Cortex Phellodendri, Chrysanthemum, Crataegus, Rubia cordifolia, Astragalus, Fructus Ligustri Lucidi, and Dioscorea opposita; the single medicine included in Comparative Prescription I includes Artemisia capillaris, Rhizoma Picrorhizoma, Rhizoma Cyperi, Artemisia annua, Centella asiatica, Radix Pseudostellariae, Radix Astragali, Piper longum, Fructus Ligustri Lucidi, Eclipta prostrata, Radix Rehmanniae Preparata, and Artificial Bezoar; Comparative Prescription II is Artemisia capillaris soup, including Artemisia capillaris, Gardenia, Rhubarb, etc. The above single medicines are extracted by ultrasonic water extraction method. The specific method is to take 10g of Chinese medicine and crush it into powder by a pulverizer. Add sterile water according to the material ratio of 1:12 to soak for 1-2h, ultrasonic extraction for 10-30min, fully stand, concentrate the supernatant at 70°C, freeze at -80°C for more than 6h, and obtain dry powder after low-temperature vacuum drying.

[0094] HepG2 cells were seeded in 384-well plates at a density of 3000 / well, and the above cell lines were cultured in an incubator for 24 hours. The culture medium was discarded, and the medium containing the same amount of single herbal extract was added to each well of the drug group; the medium containing PBS was added to the well of the control group. The above cell lines were placed in an incubator and continued to be cultured for 24 hours. Cell collection, lysis, RNA extraction, reverse transcription and library construction were carried out according to the instructions of the corresponding kits, and sequencing was performed using the Illumina HiSeq×10 platform.

[0095] (3) Enrichment of molecular signaling pathways in HepG2 cells treated with traditional Chinese medicine

[0096] After routine data quality control and sequence alignment of the sequencing in (1) above, GSEA was used to perform enrichment analysis of molecular signal pathways, and signal pathways with FDR < 0.25 and p value < 0.05 were considered as significantly enriched signal pathways. The standardized enrichment scores of the 24 pathways were calculated respectively, which were the standardized enrichment scores of traditional Chinese medicine.

[0097] (4) The improvement effects of each Chinese medicine in this patent on 24 disease signaling pathways

[0098] Depend on Figure 1 The results show that 11 Chinese medicines improve 24 disease signaling pathways in HepG2 cells. The results show that any single Chinese medicine in the composition can act on at least one of the 24 related signaling pathways. Among them, √ indicates that the pathway is improved.

[0099] Table 1 shows the improvement ratios of the three prescriptions on 24 disease signaling pathways after HEV infection of hepatocytes.

[0100] Table 1 The improvement ratio of the three prescriptions on 24 disease signaling pathways after infection of hepatocytes.

[0101]

[0102]

[0103] Example 2: Preparation of the present invention and comparative recipe

[0104] Weigh the powder according to the weight parts shown in Table 2 and mix them thoroughly. Put them into a round-bottom flask equipped with an electric heating jacket, add 10 to 15 times the volume of ultrapure water, stir and mix, heat to boiling, adjust the temperature to keep it in a slightly boiling state and continue heating for 1 to 1.5 hours, filter it through 200 mesh gauze to obtain extract 1 and save it for later use. Repeat the above steps to obtain extract 2 and save it for later use. Combine extracts 1 and 2 and use a rotary evaporator to reduce pressure and concentrate to a density of 1.1 to 1.2 g / mL (60 to 70°C), then spray dry at an inlet air temperature of 130°C, and then crush the spray-dried powder to obtain 50 to 300 mesh powder, put it into an aluminum foil bag, seal it, label it, and store it in a moisture-proof cabinet.

[0105] Table 2 Composition and mass ratio of different prescriptions

[0106]

[0107] Example 2: In vitro anti-HBV replication activity test

[0108] (1) Cytotoxicity and HBV antigen content in culture medium

[0109] HepG2.215 cells were seeded in a 384-well plate at a density of 3000 / well and placed in an incubator for 24 hours. The cell culture plate was removed from the incubator and the culture medium was slowly discarded. The culture medium containing the drug at a concentration of 1500ug / mL was added to the wells of the drug administration group, and the culture medium containing PBS was added to the wells of the control group, and the plates were placed in an incubator for another 24 hours.

[0110] Remove HepG2.215 cells from the incubator and slowly discard the culture medium. Add 110 μL of culture medium containing CCK-8 reagent. After continuing to culture for 3 hours, measure the absorbance of each well with an ELISA reader (wavelength 450 nm) and calculate the cell survival rate according to the reagent instructions.

[0111] Take out the components of the ELISA kit from the box, balance to room temperature, shake well before preparing the concentrated washing solution, and then dilute with distilled water at a ratio of 1:19. Add 50μL of negative and positive control serum or supernatant of the cell culture medium to be tested to the corresponding wells of the antibody-coated ELISA plate, then add 50μL of enzyme-labeled antibody to each well, mix well and incubate at 37℃ for 60min, balance at room temperature for 5min, wash thoroughly with washing solution 5 times, and dry after washing. Add 50μL of substrate A and B to each well, mix well and incubate at 37℃ in the dark for 15min, add 50μL of stop solution to each well and mix well, measure the OD value of each well at 450nm with an ELISA reader, and complete the measurement within 30min.

[0112] (2) Test results

[0113] For the present invention, comparative 1 and comparative 2 described in Example 3, Figure 3 As shown, at a concentration of 1500 μg / mL, the cytotoxicity of the present invention is significantly lower than that of comparative recipe 1 (p=0.0267803) and comparative recipe 2 (p=0.0016331). In the hepatitis B virus antigen detection results, the present invention showed significant efficacy in inhibiting hepatitis B virus replication at 1500 μg / mL. The inhibition of HBsAg antigen secretion was significantly better than comparative recipe 1 (p=0.03943) and comparative recipe 2 (p=0.0021411), and the inhibition of HBeAg antigen secretion was significantly better than comparative recipe 1 (p=0.0029006).

[0114] Example 3 Comparison of cytotoxicity and anti-HBV replication activity of different ratios and deletions of compound prescriptions

[0115] In order to compare the cytotoxicity and antiviral activity of the compound of the present invention at different ratios, the formula was formulated according to the weight portions shown in Table 3.

[0116] Table 3 Different proportions of compound weight

[0117]

[0118] The extracts were prepared according to the method described in Example 1, and then the cytotoxicity of each compound was tested according to the method described in Example 4. Figure 4 As shown, the three different ratios of the compound had no obvious cytotoxicity at 0-1500 μg / mL, and the three different ratios at different concentrations all had obvious inhibitory effects on the replication of hepatitis B virus, and their effects on inhibiting the secretion of HBeAg and HBsAg were similar.

[0119] Example 4 Comparison of the effects of different compositions on the improvement of disease signaling pathways in hepatitis E virus infected liver cell lines

[0120] In order to compare the improvement of the patented compound of the present invention and the different components in the compound on the related pathways after hepatitis E virus infection of liver cells, the improvement of the three compounds on 24 abnormal signal pathways after hepatitis E virus infection of liver cells were analyzed on the HepG2 cell line according to the method described in Example 1 for the compound in Table 4.

[0121] Table 4 Composition of each compound

[0122]

[0123] The improvement ratios of each compound on 24 disease signal pathways are shown in Table 5. The improvement of the 24 pathways related to hepatitis E virus infection in HepG2 cells by the present invention is better than that of the two comparison compounds. Figure 5 .

[0124] Table 5 The proportion of the number of 24 signal pathways improved by different prescriptions

[0125]

[0126] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A Chinese medicine composition, characterized in that: The invention comprises the following raw materials in parts by weight:

2. The composition according to claim 1, characterized in that The Chinese medicine composition comprises the following raw materials in parts by weight:

3. The composition according to claim 2, characterized in that The Chinese medicine composition comprises the following raw materials in parts by weight:

4. A method for preparing a Chinese medicine composition according to any one of claims 1 to 3, characterized in that: include: Mix Artemisia capillaris, Herba Artemisiae Scopariae, Polyporus, Seed of Plantago, Cortex Phellodendri, Flos Chrysanthemi, Fructus Hawthorne, Rubia cordifolia, Astragalus, Fructus Ligustri Lucidi, Dioscorea opposita and water, heat to extract, filter, concentrate and dry to obtain the product.

5. The preparation method according to claim 4, characterized in that: The mass ratio of the total amount of Artemisia capillaris, Herba Artemisiae Scopariae, Polyporus, Semen Plantaginis, Cortex Phellodendri, Flos Chrysanthemi, Fructus Crataegi, Rubia cordifolia, Radix Astragali, Fructus Ligustri Lucidi and Dioscorea opposita to water is 1:(10-15).

6. The preparation method according to claim 4, characterized in that: The temperature of the heating extraction is 100° C., the time is 1 to 1.5 hours, and the number of times of the heating extraction is 1 to 3 times.

7. The preparation method according to claim 4, characterized in that: The concentration is concentrated at 60-70° C. to 1.1-1.2 g / mL; The drying is specifically: spray drying at an inlet air temperature of 130°C; The concentration and drying process further includes crushing, wherein the crushing value is 50 to 300 meshes.

8. Use of the composition according to any one of claims 1 to 3 or the composition prepared by the preparation method according to any one of claims 4 to 7 in the preparation of drugs for treating hepatitis B virus and hepatitis E virus.

9. The use according to claim 8, characterized in that: The treatment includes improving hepatitis disease signaling pathways, reducing the secretion of hepatitis B surface antigen and hepatitis B core antigen in HepG2.215 cells, and potentially inhibiting hepatitis virus replication.

10. A product for treating hepatitis E / B virus, characterized in that: The invention comprises the composition according to any one of claims 1 to 3 or the composition prepared by the preparation method according to any one of claims 4 to 7.

11. The product according to claim 10, characterized in that The product is a medicine; the drug dosage form includes but is not limited to one or more of granules, decoctions, powders, oral liquids, tablets, pills, pastes or capsules.

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