Traditional Chinese medicine compound for improving liver regeneration microenvironment, preparation method and application

Abstract

The invention discloses a traditional Chinese medicine compound for reducing the occurrence risk of liver cancer by improving a liver regeneration microenvironment and a preparation method of the traditional Chinese medicine compound. The traditional Chinese medicine compound is prepared by the following steps: adding water into five traditional Chinese medicine decoction pieces such as mistletoe, semen cuscutae, turmeric, oldenlandia diffusa and semen coicis, decocting twice (2 hours each time), combining decoctions, filtering, standing filtrate overnight, taking supernate, concentrating into thick extract, and storing the extract for later use; taking the thick paste, adding a proper amount of starch, uniformly stirring, putting into a drying box, and drying at the temperature of below 80 DEG C; taking out the dried product, weighing, crushing, adding starch until the total weight is 450g, and uniformly mixing for later use; taking the medicinal powder, preparing a soft material by using 80% ethanol, granulating, drying at the temperature of 80 DEG C or below, and finishing granules. And taking the granules, adding 0.3% of magnesium stearate, uniformly mixing, filling into No.0 capsules, preparing 1000 granules (0.45 g per granule), carrying out aluminum-plastic plate pressing, and packaging. The compound traditional Chinese medicine is mainly used for reducing the occurrence risk of liver cancer by improving the liver regeneration microenvironment for chronic liver disease patients.

Description

Technical Field

[0001] This invention patent belongs to the field of traditional Chinese medicine, and mainly relates to a traditional Chinese medicine compound for reducing the risk of liver cancer by improving the liver regeneration microenvironment, its preparation method and application. Background Art

[0002] Liver cancer is recognized as a major and difficult disease due to its insidious onset, high incidence, high mortality, high annual growth rate, high recurrence rate and high metastasis rate. Since its exact pathogenesis is not very clear, there is currently a lack of reliable prevention and treatment measures. Previous efforts to prevent and treat liver cancer mainly focused on liver cancer cells themselves. However, in recent years, after realizing the important role of the liver cancer microenvironment in the occurrence and development of liver cancer, the concept of preventing and treating liver cancer is undergoing a major transformation. After deeply understanding the various ways in which the entire microenvironment affects the occurrence, development and metastasis of liver cancer, starting to cut off the connection between the liver cancer microenvironment and liver cancer cells is a new prevention and treatment strategy.

[0003] In recent years, with the rapid development of medical science and technology, many new discoveries have been made about the relationship between the pathogenesis of liver cancer and the liver cancer microenvironment. Among them, the mechanism research of the liver regeneration microenvironment has improved people's understanding of the pathogenesis and prevention and treatment level of liver cancer. At present, it is known that liver cancer is the result of multiple factors, multiple steps, multiple genes and multiple mutations. Although there are various hypotheses about its pathogenesis, the exact mechanism has not been clarified so far, which is the key scientific problem hindering the improvement of the prevention and treatment level. Researchers have been trying to clarify the occurrence mechanism and cell origin of liver cancer. In recent years, the theory of cancer stem cells has attracted much attention. It is believed that tumors may be caused by the interaction of the cell microenvironment, carcinogens and genetic factors on normal stem cells during the long-term self-renewal process, resulting in genetic or epigenetic changes in some molecules in the growth and differentiation regulation pathway, leading to abnormal expression, and then causing excessive abnormal proliferation of cells. Tumors are not only a genetic disease, but also a stem cell disease. Gene mutations act on stem cells, and stem cells mutate into cancer stem cells, which is the key to the occurrence, regeneration, metastasis and recurrence of tumors. The latest research shows that primary liver cancer may be the result of the out-of-control of liver stem cells during self-renewal, leading to undifferentiated or incompletely differentiated states. Epigenetic changes in liver stem cells may be one of the important mechanisms, and the abnormal liver regeneration microenvironment plays a crucial role in this process. The liver regeneration microenvironment of liver cancer is different from the normal liver regeneration microenvironment, mainly manifested as abnormally expressed liver regeneration regulatory factors and their related receptors, disordered activation or inactivation of signaling pathways, and disordered organizational structure changes, etc., collectively referred to as abnormal or deteriorated liver regeneration microenvironment. The normal liver regeneration microenvironment delays, prevents or even reverses the occurrence, development or recurrence and metastasis of liver cancer, while the abnormal or deteriorated liver regeneration microenvironment initiates, promotes or even accelerates the occurrence, development or recurrence and metastasis of liver cancer.

[0004] Studies have shown that the abnormal liver regeneration microenvironment formed by liver inflammation and fibrosis plays an important role in the occurrence and development of liver cancer. Approximately 80% of liver cancers occur against the background of chronic inflammation and fibrosis caused by chronic viral hepatitis. The treatment and prognosis of liver cancer are also complicated by different tumor stages and degrees of liver function abnormalities. Most chronic liver diseases are characterized by diffuse chronic inflammation, necrosis, and fibrosis. The occurrence of chronic inflammation and fibrosis is a dynamic process involving the secretion and paracrine interactions of accumulated lymphocytes, macrophages, and stromal cells. Inflammatory cells belonging to both innate and adaptive immunity are involved in liver injury and liver regeneration. The imbalance between liver injury and liver regeneration determines the occurrence and development of liver fibrosis. Damaged hepatocytes, Kupffer cells, and hepatic stellate cells are all involved in inflammation induction. The interaction between the stromal microenvironment, stroma, and infiltrating lymphocytes not only determines the occurrence and development of fibrosis in chronic inflammatory liver diseases but also affects the occurrence and development of liver cancer. It has been proven that the disordered liver regeneration process in patients with chronic viral hepatitis produces a large amount of transforming growth factor-β1 (TGF-β1), which is a key factor leading to the abnormal regenerative microenvironment of liver fibrosis affecting the occurrence and development of liver cancer. First, the TGF-β / Smads signaling pathway is disrupted, the cell growth cycle is disordered, and hepatocytes become cancerous. Second, due to the reduced expression of the TGF-β receptor in liver cancer cells, the inhibitory effect on the proliferation of potentially residual liver cancer cells is lost. Third, due to the immunosuppressive effect of high-concentration TGF-β1 and its role in promoting angiogenesis in the body, a tissue microenvironment conducive to the occurrence or recurrence of liver cancer is created. In recent years, with the deepening of the understanding of the existence and importance of stem cells and the stem cell niche, the regulatory role of the liver regeneration microenvironment in the activation, proliferation, and differentiation of oval cells has received extensive attention. The deterioration of the hepatic stem cell niche is the liver regeneration microenvironment for its transformation into liver cancer stem cells. The loss of temporality and regulation in the regeneration and healing process after chronic inflammation and injury may lead to liver cancer. Malignant transformed hepatocytes can replace the atypical hyperplastic nodules in the regeneration process, and the liver cancer stem cells present therein determine the occurrence and development of liver cancer. Hypoxia and inflammation are also the main factors that stimulate angiogenesis, which can promote the growth of liver cancer. Inflammatory-related signals such as Toll-like receptor (TLR) 4 and nuclear factor (NF)-κB can promote the proliferation and migration of liver cancer cells by producing a large number of cytokines, chemotactic and growth factors, angiogenesis and lymphangiogenesis factors, and altering the stroma.

[0005] Therefore, the disorder of the liver regeneration microenvironment is one of the necessary conditions for the occurrence and development of liver cancer. Exploring the relevant mechanisms of the influence of the liver regeneration microenvironment on the occurrence and development of liver cancer and proposing measures and programs to prevent and treat the occurrence and development of liver cancer by improving the liver regeneration microenvironment of liver cancer are of great significance for improving the prevention and treatment ability and level of liver cancer. It is urgent to research and develop effective traditional Chinese medicine preparations that can reduce the risk of liver cancer by improving the liver regeneration microenvironment. Summary of the Invention

[0006] The present invention provides a traditional Chinese medicine compound for improving the hepatic regeneration microenvironment and reducing the risk of liver cancer, a preparation method thereof, and an application thereof in the preparation of a drug for improving the hepatic regeneration microenvironment and reducing the risk of liver cancer. The prescription of the traditional Chinese medicine compound is derived from the clinical experience formula of Chief Physician Li Hanmin of the Institute of Liver Diseases of Hubei Provincial Hospital of Traditional Chinese Medicine for many years. His research team has developed a traditional Chinese medicine compound for reducing the risk of liver cancer by improving the hepatic regeneration microenvironment. The traditional Chinese medicine compound is made from the following raw materials in parts by weight: 1-30 parts of mistletoe, 1-30 parts of dodder, 1-10 parts of turmeric, 1-50 parts of oldenlandia diffusa, and 1-60 parts of coix seed. A traditional Chinese medicine compound preparation, which includes the above traditional Chinese medicine compound and pharmaceutically acceptable excipients. Preferably, the traditional Chinese medicine compound is made from the following raw materials in parts by weight: 7-30 parts of mistletoe, 5-25 parts of dodder, 5-10 parts of turmeric, 7-40 parts of oldenlandia diffusa, and 7-40 parts of coix seed. More preferably, the traditional Chinese medicine compound is made from the following raw materials in parts by weight: 7-15 parts of mistletoe, 5-10 parts of dodder, 5-10 parts of turmeric, 7-15 parts of oldenlandia diffusa, and 7-15 parts of coix seed.

[0007] A traditional Chinese medicine compound preparation, which includes the above traditional Chinese medicine compound and pharmaceutically acceptable excipients.

[0008] The preparation method of the traditional Chinese medicine compound preparation is as follows: Take the five traditional Chinese medicine pieces of mistletoe, dodder, turmeric, oldenlandia diffusa, and coix seed according to parts by weight, decoct them twice with water for 2 hours each time, combine the decoctions, filter, let the filtrate stand overnight, take the supernatant, concentrate it to a thick extract, and store the extract for later use; Take the above thick extract, add an appropriate amount of starch, mix evenly, place it in a drying oven, and dry it at a temperature below 80°C; Take out the dried product, weigh it, crush it, and then add starch to make the total weight reach 450 g, mix evenly, and set aside; Take the above medicinal powder, make soft materials with 80% ethanol, granulate, dry it at a temperature below 80°C, and size the granules; Take the above granules, add 0.3% magnesium stearate, mix evenly, and fill them into capsules.

[0009] In the traditional Chinese medicine compound of the present invention, mistletoe and dodder supplement the liver and kidney, turmeric promotes blood circulation to remove stasis, dredges collaterals and softens hardness, and oldenlandia diffusa and coix seed promote diuresis and detoxify. Together, they play the roles of tonifying the kidney and liver, activating blood circulation and dredging collaterals, softening hardness and detoxifying.

[0010] Mistletoe is bitter and sweet in taste and neutral in nature. It belongs to the liver and kidney meridians. Modern pharmacological studies have confirmed that in addition to having good pharmacological effects on the cardiovascular system, it also has the effects of improving microcirculation, antiplatelet aggregation, and anti-tumor. We have achieved good effects in the treatment of acute and chronic liver diseases.

[0011] Cuscuta chinensis is pungent, sweet, and neutral in nature, entering the liver and kidney meridians. It has the effects of tonifying the kidney and replenishing essence, nourishing the liver and improving eyesight, and preventing miscarriage. Experiments have shown that: when the aqueous decoction of Cuscuta chinensis is administered intragastrically to "yang-deficient" mice, it can significantly increase their body weight, kidney weight, thymus weight, white blood cell count, red blood cell count, hemoglobin, and the activity of superoxide dismutase; when administered intragastrically to male mice, it can significantly extend the swimming time of the mice and their survival time under hypoxic conditions, and also enhance the non-specific resistance of the mice. Cuscuta chinensis can increase the luteinizing function of the hypothalamus-pituitary-ovary axis. This effect is not due to directly stimulating the secretion of pituitary luteinizing hormone, but rather due to enhancing the responsiveness of the pituitary to LRH and the ovary to LH. The hot water extract of Cuscuta chinensis can significantly delay the growth of skin tumors and the spread of malignant tumors induced by 7,12-dimethylbenz[a]anthracene (DMBA) in mice. The ethanol-water extract of Cuscuta chinensis can significantly inhibit the incorporation of tritiated thymidine (3H-TdR) into the DNA synthesis of Ehrlich ascites carcinoma cells, suggesting its anti-cancer effect. The ethanol extract of Cuscuta chinensis has the effect of promoting the growth of granulocyte progenitor cells.

[0012] Curcuma longa is pungent, bitter, and warm in nature, entering the spleen and liver meridians. Its functions include promoting blood circulation to remove blood stasis, regulating qi movement, and dredging channels to relieve pain. It has various pharmacological effects such as antiviral, lipid-lowering, anti-inflammatory, cholagogic, antioxidant, and anti-tumor effects.

[0013] Oldenlandia diffusa is bitter, tasteless, and cold in nature. Its main effects are clearing heat and detoxifying, dissipating stasis and resolving masses, and promoting diuresis and removing dampness. It is especially good at treating various types of inflammation. In clinical practice, it has been found that when Oldenlandia diffusa is properly combined, it can treat a variety of diseases and is widely used in acute and chronic liver diseases and tumor diseases.

[0014] Coix lacryma-jobi: It is sweet, tasteless, and cool in nature, entering the spleen, stomach, and lung meridians. It has the functions of strengthening the spleen and promoting diuresis, clearing heat and discharging pus, relieving obstruction and promoting diuresis. It has various pharmacological effects such as anti-tumor, enhancing immunity (coix seed oil can promote cellular immunity and humoral immunity), and anti-inflammatory effects.

[0015] Advantages of the present invention: The traditional Chinese medicine compound described in the present invention has the effects of tonifying the kidney and nourishing the liver, promoting blood circulation to dredge collaterals, softening hardness and detoxifying, and has the function of regulating the liver regeneration and repair mechanism. It is an effective preparation for normalizing liver regeneration and repair after liver injury, and is mainly used for treating patients with chronic liver diseases who present with symptoms such as liver and kidney deficiency, blood stasis blocking collaterals, and toxins forming masses. It can reduce the risk of liver cancer in patients with chronic liver diseases by anti-liver fibrosis and improving the microenvironment of liver regeneration. Brief Description of the Drawings

[0016] Figure 1 It is a process diagram for preparing the traditional Chinese medicine compound capsule in Example 1 provided by the present invention.

[0017] The realization of the object, functional characteristics, and advantages of the present invention will be further described in conjunction with the embodiments and with reference to the attached drawings. Detailed Embodiments

[0018] Example 1: Traditional Chinese Medicine Compound Capsule

[0019] I. Prescription Composition

[0020]

[0021] II. Main Raw and Auxiliary Materials Specifications and Quality Standards for Preparation Process

[0022] The names, specifications and standards of the main raw and auxiliary materials are shown in Table 1.

[0023] Table 1 Names, Specifications and Standards of Main Raw and Auxiliary Materials

[0024]

[0025] III. Preparation Process

[0026] 1. Extraction Process

[0027] The above five flavors are decocted twice with water for 2 hours each time. The decoction liquids are combined, filtered, and the filtrate is allowed to stand overnight. The supernatant is taken and concentrated to a thick extract, which is stored for later use.

[0028] 2. Drying Process

[0029] Take the above-mentioned thick extract, add an appropriate amount of starch, mix evenly, place it in a drying oven, and dry it at a temperature below 80°C; take out the dried product, weigh it, crush it, and then add starch to make the total weight reach 450 g, mix well, and store for later use.

[0030] 3. Granulation

[0031] Take the above-mentioned medicinal powder, prepare soft materials with 80% ethanol, granulate, dry at a temperature below 80°C, and screen the granules.

[0032] 4. Sub-packaging and Packaging

[0033] Take the above-mentioned granules, add 0.3% magnesium stearate, mix well, fill into No. 0 capsules to make 1000 capsules (each capsule is 0.45 g), and package with aluminum-plastic pressing plates to obtain the product.

[0034] The preparation process diagram is shown in the appendix Figure 1 .

[0035] Example 2: Traditional Chinese Medicine Compound Tablets

[0036] Prescription Composition: Mistletoe 600 g, Cuscuta chinensis 200 g, Turmeric 200 g, Oldenlandia diffusa 600 g, Coix seed 600 g.

[0037] Preparation method: The above five kinds of medicinal slices are decocted in water twice, each time for 2 hours. The decoctions are combined, filtered, and the filtrate is allowed to stand overnight. The supernatant is taken, concentrated into a thick extract, and the extract is stored for later use. The extract is mixed with an appropriate amount of dextrin, stirred evenly, dried, pulverized, and sieved. The obtained dry extract powder is granulated with an appropriate amount of ethanol, sized, and an appropriate amount of magnesium stearate is added and pressed into tablets.

[0038] Example 3: Traditional Chinese medicine compound granule

[0039] Prescription composition: 900 g of Taxillus chinensis, 500 g of Cuscuta chinensis, 500 g of Curcuma longa, 700 g of Hedyotis diffusa, 700 g of Coix lacryma-jobi.

[0040] The above five flavors are decocted in water twice, each time for 2 hours. The decoctions are combined, filtered, and the filtrate is allowed to stand overnight. The supernatant is taken, filtered, and concentrated into a clear extract with a relative density of 1.25 - 1.30 (60 °C). An appropriate amount of dextrin is added, granulated, dried, and made into granules.

[0041] Example 4: Randomized controlled clinical trial on the prevention of liver cancer by a traditional Chinese medicine compound capsule for liver cancer in reducing the risk of liver cancer in patients with chronic hepatitis B

[0042] Some studies have shown that the abnormal liver regeneration microenvironment formed by liver inflammation and fibrosis plays an important role in the occurrence and development of liver cancer. Approximately 80% of liver cancers occur against the background of chronic inflammation and fibrosis caused by chronic viral hepatitis. The treatment and prognosis of liver cancer are also complicated due to different tumor stages and degrees of liver function abnormalities. Liver fibrosis is one of the main forms of abnormal liver regeneration. Preventing and treating the occurrence and development of liver cancer by improving the liver regeneration microenvironment through anti-liver fibrosis is a new prevention and treatment strategy. However, there is currently a lack of traditional Chinese medicine preparations that can prevent and treat the occurrence and development of liver cancer by improving the liver regeneration microenvironment. This clinical study conducted a randomized controlled observation on the clinical efficacy of the capsule of the traditional Chinese medicine compound in Example 1 (Taxillus chinensis, Cuscuta chinensis, Curcuma longa, Hedyotis diffusa, Coix lacryma-jobi) (hereinafter referred to as: traditional Chinese medicine compound capsule for liver cancer) in reducing the risk of liver cancer in patients with chronic hepatitis B by anti-liver fibrosis and improving the liver regeneration microenvironment. The relatively complete clinical data collected are summarized as follows.

[0043] 1 Data and methods

[0044] 1.1 Case selection: 121 patients with chronic hepatitis B who visited the outpatient department or were hospitalized in Hubei Provincial Hospital of Traditional Chinese Medicine from January 2008 to December 2010 were selected. They were randomly divided into a treatment group of 61 cases and a control group of 60 cases. There were no significant differences in age, disease course, and severity of the disease between the two groups of cases (P>0.05), and the clinical observation and research were comparable. 1.2 Diagnostic criteria: The Western medicine diagnostic criteria refer to the diagnostic criteria of viral hepatitis jointly revised by the Infectious Diseases and Parasitology Branch and the Liver Diseases Branch of the Chinese Medical Association in 2000; the traditional Chinese medicine diagnostic criteria refer to the "Clinical Guidelines for the Treatment of Viral Hepatitis with Traditional Chinese Medicines - Syndrome of Deficiency of the Liver and Kidney" of the Ministry of Health of the People's Republic of China and the "Diagnostic Criteria for Traditional Chinese Medicine of Viral Hepatitis (Trial)" adopted by the Tianjin Conference of the Professional Committee of Internal Medicine of Traditional Chinese Medicine of the Chinese Association of Traditional Chinese Medicine in December 1991.

[0045] 1.2.1 Western medicine diagnostic criteria for chronic hepatitis B: Those with a previous history of positive HbsAg, a disease course of more than half a year, and still having symptoms, signs, and abnormal liver function, and positive hepatitis B virus markers, with HBV-DNA greater than or equal to 10 4 copies / ml, and those with pathological histological changes can all be diagnosed as chronic hepatitis B. Some cases have an unclear onset time, but those with imaging, pathological examinations, or liver function damage consistent with chronic hepatitis changes can also be used for diagnosis.

[0046] To comprehensively reflect the degree of liver damage, the diagnostic criteria for chronic hepatitis B can be clinically divided into:

[0047] Mild: The disease course is short, the symptoms are not obvious, or although there are symptoms and signs, only 1 or 2 biochemical indicators are mildly abnormal;

[0048] Moderate: The symptoms, signs, and laboratory examinations are between mild and severe;

[0049] Severe: There are obvious or persistent hepatitis symptoms, such as fatigue, poor appetite, abdominal distension, loose stools, etc., and may be accompanied by a liver disease face, liver palms, spider nevi, or hepatosplenomegaly, excluding other causes and without portal hypertension. Laboratory examinations show repeated or persistent elevation of ALT, decreased albumin, or inversion of the albumin / globulin ratio.

[0050] 1.3 Quantification scoring method for traditional Chinese medicine syndromes

[0051] The quantification scoring method for traditional Chinese medicine syndromes is shown in Table 2

[0052] Table 2

[0053]

[0054]

[0055] 1.4 Inclusion and exclusion criteria

[0056] (1) Inclusion criteria:

[0057] ①Voluntarily serve as a subject and be able to accept the investigational drug, and ensure to complete the treatment course;

[0058] ②Aged 18 - 65 years old at the time of enrollment;

[0059] ③The clinical diagnosis of the clinical trial conforms to the clinical diagnostic criteria for chronic hepatitis B liver fibrosis;

[0060] ④Stop taking Chinese and Western medicines with the above-mentioned diseases as the main indications and other treatment methods for the above-mentioned diseases two weeks before the observation.

[0061] (2) Exclusion criteria:

[0062] ①Under 18 years old or over 65 years old;

[0063] ②Co-infected with other hepatitis viruses (such as hepatitis A, C, D, E) etc.;

[0064] ③Combined with organic diseases of important organs such as cardiovascular, lung, and kidney, severe primary diseases of the hematopoietic system, and psychiatric patients;

[0065] ④Those with complications such as spontaneous or tuberculous peritonitis, upper gastrointestinal bleeding, and hepatic encephalopathy;

[0066] ⑤Pregnant and lactating women and those allergic to this medicine.

[0067] 1.5 Treatment method All patients in the treatment group orally took the anti - liver cancer traditional Chinese medicine compound capsule (mistletoe, dodder seed, turmeric, oldenlandia diffusa, coix seed) supervised by the Institute of Liver Diseases of the Affiliated Hospital of Hubei College of Traditional Chinese Medicine, 6 capsules each time, 3 times a day, taken with warm boiled water, and the treatment course was 3 months. The control group used Yiganling capsules, 4 capsules each time, 3 times a day, taken with warm boiled water, and the treatment course was 3 months.

[0068] 1.6 Observation indicators and detection methods Clinical symptoms and signs adopted the conventional syndrome differentiation criteria, and some signs were combined with modern diagnosis and treatment techniques and methods (such as liver and spleen enlargement mainly referring to the B - ultrasound detection results, and jaundice mainly referring to the serum bilirubin detection results). Conventional liver function indicators: serum bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total serum protein (TB), serum albumin (HB), etc., were detected by an automatic biochemical analyzer. Hepatitis B virus markers (HBV - M): HbsAg, HbsAb, HbeAg, HbeAb, HbcAb, were detected by the ELASA method. HBV - DNA: was detected by the dot blot hybridization method. Serum liver fibrosis indicators: serum hyaluronic acid (HA), serum type Ⅲ procollagen (PCⅢ), were detected by radioimmunoassay. It was examined at least once before and after treatment.

[0069] 1.7 The comprehensive curative effect was judged with reference to the "Trial Criteria for Judging the Curative Effect of Traditional Chinese Medicine in Viral Hepatitis" formulated at the Tianjin Conference of the Professional Committee of Internal Medicine of Traditional Chinese Medicine of the Chinese Association of Traditional Chinese Medicine in 1991 and the criteria for evaluating the curative effect of new traditional Chinese medicines in the guiding principles for clinical research on viral hepatitis revised by the Pharmaceutical Administration of the Ministry of Health [1]. The observed cases were followed up for more than 1 year, and the curative effect was judged according to the following criteria: ① Clinical cure: The main symptoms disappeared, the hepatosplenomegaly disappeared or regressed, there was no obvious percussion pain in the liver area, and the liver function returned to normal; HbeAg and ABV-DNA turned negative for more than 1 year, and the serum liver fibrosis indexes returned to normal; ② Marked effect: The main symptoms basically disappeared, the liver and spleen regressed and there was no obvious tenderness, the liver function basically returned to normal, the serum liver fibrosis indexes decreased by more than 2 / 3, but there was no obvious change in the hepatitis B virus markers; ③ Effective: The main symptoms improved or disappeared, the hepatosplenomegaly regressed, the liver function ALT and AST decreased by 1 / 2, and the serum liver fibrosis indexes decreased by 1 / 2, but there was no obvious change in the hepatitis B virus markers; ④ Ineffective: All indexes showed no change or deterioration, and the clinical symptoms showed no change or aggravation.

[0070] 1.8 Liver cancer risk assessment There are various assessment models for the risk of liver cancer in patients with chronic hepatitis B. In this study, the REACH-B liver cancer risk prediction model widely used in the international academic community and the LI-MA liver cancer prediction model established by ourselves were adopted.

[0071] 1.8.1 REACH-B liver cancer risk prediction model (Hwai-I Yang, Morris Sherman, Jun Su. Nomograms for Risk of Hepatocellular Carcinoma in Patients With Chronic hepatitis B Virus Infection. JOURNAL OF CLINICAL ONCOLOGY, 2010;28(14):2437-2444. Huai-I Yang, Man-Fung Yuen, Henry Lik-Yuen Chan. Risk estimation for hepatocellular carcinoma in chronic hepatitis B (REACH-B): development and validation of a predictive score. Lancet Oncol, 2011;12:568–574.)

[0072] The parameters of the REACH-B liver cancer prediction model are based on 5 observation indicators such as the patient's gender, age, ALT level, HBeAg positivity, and HBV-DNA level. Weights are compared according to the standardized regression coefficients of the observation indicators, and quantitative scores are assigned to the above variables. Age 34 and below is recorded as 0 points, and 35-39 years old, 40-44 years old, 45-49 years old, 50-54 years old, 55-59 years old, and 60-65 years old are recorded as 1, 2, 3, 4, 5, and 6 points respectively. Females are recorded as 0 points, and males are recorded as 2 points. According to the ALT level, it is divided into three grades: <15, 15-44, and 45 and above, which are recorded as 0, 1, and 2 points respectively. According to the HBV-DNA load, <300 and 300-9999 are recorded as 0 points, 10000-99999 is recorded as 3 points, 100000-999999 is recorded as 5 points, and 1000000 is recorded as 4 points. The total REACH-B score ranges from 0 to 17 points, and the total score is the score used to predict the level of risk. The higher the score, the greater the likelihood of developing liver cancer. The specific scoring is shown in Table 3.

[0073] Table 3: Scoring Criteria for the REACH-B Liver Cancer Risk Prediction Model

[0074]

[0075] 1.8.2 LI-MA Chronic Hepatitis B Liver Cancer Occurrence Risk Prediction Model (Li Hanmin. Basics and Clinical Practice of Traditional Chinese Medicine in Regulating Liver Regeneration [M]. Huazhong University of Science and Technology Press, the 1st edition in December 2016, 1-644.)

[0076] The LI-MA chronic hepatitis B liver cancer occurrence risk prediction model selects risk factors affecting the occurrence of liver cancer in chronic hepatitis B, including 5 observation indicators such as age, gender, TBIL, GGT, and HBV-DNA. Weights are compared according to the standardized regression coefficients of the observation indicators, and quantitative scores are assigned to the above variables. Age 50 and below is recorded as 0 points, and over 50 years old is recorded as 3 points. Females are recorded as 0 points, and males are recorded as 3 points. According to the GGT level, it is divided into two grades: <55 and ≧55, which are recorded as 0 and 17 points respectively. According to the TBIL level, it is divided into two grades: <18 and ≧18, which are recorded as 0 and 12 points respectively. According to the HBV-DNA load, <1000, 1000-99999 is recorded as 2 points, and ≧100000 is recorded as 5 points. The total score ranges from 0 to 41 points. The total score of the above 5 variables is the score used to predict the level of risk. The higher the score, the higher the risk of developing serious clinical outcomes. 7 and 19 are the optimal cut-off points of the prediction scoring model, that is, "≤7 = low risk; 7-19 = medium risk; >19 = high risk". The specific scoring is shown in Table 4.

[0077] Table 4: Scoring Criteria for the LI-MA Liver Cancer Risk Prediction Model

[0078]

[0079] 1.9 Safety and Other Evaluation Criteria

[0080] 1.9.1 Safety Observation Indicators

[0081] (1) General physical examination items;

[0082] (2) Routine blood, urine, and feces tests;

[0083] (3) Electrocardiogram, X-ray, liver function (ALT, AST), and renal function (BUN, Cr) examinations.

[0084] 1.9.2 Adverse Reaction Evaluation and Judgment Criteria

[0085] (1) Adverse reaction evaluation criteria: A. The time of onset of the adverse reaction coincides with the medication time. B. The adverse reaction is related to the known adverse reactions of the drug. C. The adverse reaction cannot be explained by other reasons. D. The adverse reaction disappears after drug withdrawal. E. The adverse reaction reappears after drug administration.

[0086] (2) Judgment criteria for the relationship between adverse events and the investigational drug: The researcher should evaluate the possible association between adverse events, the investigational drug, and concomitant medications, and rate them according to the following 5 groups of classification criteria.

[0087] ① Definitely related: The reaction occurs in a reasonable time sequence after medication, and the reaction conforms to the known reaction type of the suspected drug; it improves after drug withdrawal and reappears after repeated administration.

[0088] ② Probably related: The reaction occurs in a reasonable time sequence after medication, and the reaction conforms to the known reaction type of the suspected drug; the patient's clinical condition or other treatment methods may also produce this reaction.

[0089] ③ Probably unrelated: The reaction does not occur in a very reasonable time sequence after medication, and the reaction does not very conform to the known reaction type of the suspected drug; the patient's clinical condition or other treatment methods may produce this reaction.

[0090] ④ Unrelated: The reaction does not occur in a reasonable time sequence after medication, and the reaction conforms to the known reaction type of non-experimental drugs; the patient's clinical condition or other treatment methods may produce this reaction, the disease condition improves or the reaction disappears after stopping other treatment methods, and the reaction reappears after repeated use of other treatment methods.

[0091] ⑤ Unable to evaluate: The reaction has no clear relationship with the medication time and is similar to the known reaction type of the drug; other medications used simultaneously may also produce the same reaction.

[0092] 19.3 Serious Adverse Events

[0093] (1) Serious Adverse Event Judgment: An adverse event is classified as a serious adverse event (SAE) when it meets one or more of the following criteria:

[0094] ① Death

[0095] ② Life-threatening (e.g., at immediate risk of death)

[0096] ③ Resulting in hospitalization or prolonged hospital stay

[0097] ④ Permanent or severe disability

[0098] ⑤ Congenital malformation

[0099] Some medical events that have not resulted in death, life-threatening situations, or the need for hospitalization, but are considered by appropriate medical judgment to be likely to cause harm to the patient or subject, or require medical or surgical treatment to avoid the above situations, should also be regarded as serious adverse events.

[0100] (2) Recording and Reporting of Serious Adverse Events

[0101] Any serious adverse event during the clinical trial must be immediately reported to the institution and the main research institution, as well as the ethics committee of the clinical research base. At the same time, the researcher must fill out the serious adverse event form, recording the occurrence time, severity, duration, measures taken, and outcome of the serious adverse event.

[0102] 1.9.4 Safety Evaluation Criteria

[0103] Grade 1: Safe, without any adverse reactions. That is, no adverse reactions were found during the observation, and all safety indicators were normal before and after drug administration.

[0104] Grade 2: Relatively safe, that is, there are mild adverse reactions, but no treatment is required and drug administration can continue; all safety indicators are normal before and after drug administration.

[0105] Grade 3: There are safety issues, that is, there are moderate adverse reactions, and drug administration can continue after treatment. Or / and one safety indicator is slightly abnormal, but it can recover after drug withdrawal.

[0106] Grade 4: Poor safety. That is, there are moderate to severe adverse reactions, the patient cannot tolerate them and stops taking the drug on their own; or / and one or more safety indicators are abnormal.

[0107] 1.9.5 Case Withdrawal

[0108] (1) Judgment of Withdrawal: All patients who have signed the informed consent form and are screened and qualified to enter the trial have the right to withdraw from the clinical trial at any time. Regardless of when and for what reason they withdraw, as long as they have not completed the full-course observation of the clinical trial, they are considered withdrawn cases.

[0109] (2) Common reasons for dropout:

[0110] ① Adverse events

[0111] ② Lack of efficacy

[0112] ③ Violation of the trial protocol (including poor compliance)

[0113] ④ Loss to follow-up (including patients withdrawing on their own)

[0114] ⑤ Others

[0115] (3) Handling of dropout cases: When a patient drops out, the researcher must fill in the reason for dropout in the CRF, contact the patient as much as possible, complete all the assessable items, fill in the treatment non-follow-up record form, and record the time of the last medication as accurately as possible. For those who drop out due to adverse reactions, the relationship with the trial drug must be judged through follow-up and recorded in the CRF.

[0116] 2 Statistical methods: For measurement data, t-test is used; for count data, χ2 test and Ridit analysis are used.

[0117] 3 Results

[0118] 3.1 Changes in serum liver fibrosis markers before and after treatment

[0119] The normal reference value of PCⅢ is < 120 ng / L; the normal reference value of HA is < 110 ng / L. Before treatment, the levels of serum liver fibrosis markers PCⅢ and HA in the treatment group and the control group, P > 0.05; after treatment, the levels of PCⅢ and HA in the treatment group were significantly lower than those in the control group, P < 0.01; see Table 1.

[0120] Table 1 Changes in serum PCⅢ and HA before and after treatment (x±s)

[0121]

[0122] Compared with the control group, △P < 0.01.

[0123] 3.2 Changes in liver function ALT and AST after treatment

[0124] Before treatment, ALT in all cases of both groups had varying degrees of abnormality. After treatment, the normalization rate in the treatment group was 62.3%; the normalization rate in the control group was 33.33%. When compared between the two groups, P < 0.05, showing a significant difference.

[0125] Before treatment, AST in all cases of both groups had varying degrees of abnormality. After treatment, the normalization rate in the treatment group was 54.10%. The normalization rate in the control group was 26.66%. When compared between the two groups, P < 0.05, showing a significant difference. See Table 2.

[0126] Table 2 Changes in ALT and AST before and after treatment (normalization rate) (%)

[0127] Group ALT AST Control group 33.33(20 / 60) 26.66(16 / 60) Treatment group 62.30(38 / 61)△ 54.10(33 / 61)△

[0128] Compared with the control group, △P < 0.05.

[0129] 3.3 Changes in hepatitis B virus indicators before and after treatment

[0130] The negative conversion rate of HbeAg in the treatment group was 26.22%; the negative conversion rate of HBV-DNA was 24.59%. In the control group, they were 13.33% and 11.66% respectively. There was a significant difference between the two groups, P < 0.05. See Table 3.

[0131] Table 3 Negative conversion changes in hepatitis B virus indicators before and after treatment (%)

[0132] Group HbsAg HbeAg HBV-DNA Control group 6.66(4 / 60) 13.33(8 / 60) 11.66(7 / 60) Treatment group 8.19(5 / 62) 26.22(16 / 61)△ 24.59(15 / 61)△

[0133] Compared with the control group, △P < 0.05.

[0134] 3.4 Improvement of syndromes before and after treatment

[0135] After 3 months of treatment, the symptoms of hypochondriac pain, soreness and weakness of the waist and knees, insomnia and dreaminess, dizziness and tinnitus, feverish palms and soles or low fever, dry throat and mouth in the treatment group were significantly improved compared with the control group (P < 0.05); there were significant differences in the improvement of tongue and pulse conditions compared with the control group (P < 0.05); the jaundice subsided, with a significant difference compared with the control group (P < 0.05). See Table 4.

[0136] Table 4 Changes in the quantitative scores of TCM syndromes in the two groups

[0137]

[0138] 3.5 Determination of comprehensive efficacy

[0139] According to the above comprehensive efficacy determination criteria, in the treatment group, 13 cases (21.3%) were clinically cured, 22 cases (36.1%) showed marked efficacy, 21 cases (34.4%) were effective, and 5 cases (8.2%) were ineffective. The total effective rate was 91.8%; in the control group, 4 cases (6.6%) were clinically cured, 11 cases (18.3%) showed marked efficacy, 21 cases (35%) were effective, and 25 cases (41.6%) were ineffective. The total effective rate was 59.9%; the efficacy of the treatment group was better than that of the control group. After statistical processing, the difference was significant, P < 0.05.

[0140] Table 5 Comparison of comprehensive efficacy between the two groups (%)

[0141] Group Cured Markedly effective Effective Ineffective Total effective Control group 6.6 18.3 35.0 41.6 59.9 Treatment group 21.3 36.1 34.4 8.2 91.8△

[0142] When comparing the treatment group with the control group, △P < 0.05.

[0143] 3.6 Evaluation of the risk of liver cancer reduction by a traditional Chinese medicine compound preparation for preventing liver cancer using the REACH - B model

[0144] At the time of enrollment, the age, gender, ALT level, DNA load, and whether the e - antigen was negative of each group of patients were analyzed according to the REACH - B liver cancer risk prediction model: The ages of the patients in the treatment group of the traditional Chinese medicine compound preparation for preventing liver cancer and the control group were 41.35 ± 10.48 and 40.98 ± 8.76 respectively. After statistical comparison between groups, the difference was not significant, P > 0.05. After statistical comparison of the male - female ratio between groups, the difference was not significant, P > 0.05. The ALT levels of each group of patients at the time of enrollment were 38.43 ± 19.12 and 40.23 ± 22.38 respectively. After statistical comparison between groups, there was no obvious difference, P > 0.05. The virus loads of HBV - DNA - positive patients in each group were 2.69×10 6 ±1.64×10 6 、3.43×10 6 ±1.25×10 6 , After statistical comparison between groups, the difference was not significant, P > 0.05.

[0145] At the time of enrollment, each person in each group was given a REACH - B score. At baseline, the REACH - B scores of the treatment group of the traditional Chinese medicine compound preparation for preventing liver cancer and the control group were 6.42 ± 2.12 and 6.53 ± 3.19 respectively. After statistical comparison between groups, the difference was not significant, P > 0.05. After 6 and 12 months of treatment, the scores of the treatment group of the traditional Chinese medicine compound preparation for preventing liver cancer were (4.31 ± 1.76, 3.67 ± 1.82) respectively, and those of the control group were (6.62 ± 1.12, 6.25 ± 1.21) respectively. After statistical treatment, the difference was significant, P < 0.05. Compared with before treatment, the REACH - B scores of each group decreased significantly compared with before treatment, P < 0.05. The results show that the traditional Chinese medicine compound preparation for preventing liver cancer has the effect of reducing the risk of liver cancer in patients with chronic hepatitis B.

[0146] 3.7 Evaluation of the risk of liver cancer reduction by "Diwu Yanggan Capsule" using the LI - MA model

[0147] At the time of enrollment, the age, gender, GGT level, TBIL level, and DNA load of the patients in the traditional Chinese medicine compound preparation treatment group and the control group for preventing liver cancer were analyzed according to the LI-MA prediction model for the risk of liver cancer in chronic hepatitis B. After statistical processing of the comparison between groups, the differences were not significant, P>0.05. At baseline, the LI-MA scores of the traditional Chinese medicine compound preparation treatment group and the control group for preventing liver cancer were 8.34±1.23 and 9.29±1.14 respectively. After statistical processing of the comparison between groups, the differences were not significant, P>0.05. After 6 and 12 months of treatment, the scores of the traditional Chinese medicine compound preparation treatment group for preventing liver cancer were (4.83±1.73, 5.21±1.16) respectively, and those of the control group were (7.98±2.57, 8.12±2.54). After statistical processing, the differences were significant, P<0.05. Compared with before treatment, the MA scores of each group were significantly decreased compared with before treatment, P<0.05. The results showed that the traditional Chinese medicine compound preparation for preventing liver cancer had the effect of reducing the risk of liver cancer in patients with chronic hepatitis B.

[0148] According to the high-risk (score>19), medium-risk (score = 7-19), and low-risk (score≤7) criteria established based on the LI-MA scoring of the risk prediction model for liver cancer in chronic hepatitis B, the "high / medium / low" composition ratios at the time of enrollment were 39 / 16 / 6 in the traditional Chinese medicine compound preparation treatment group for preventing liver cancer and 40 / 15 / 5 in the control group. After statistical comparison of the number of people in each risk level between groups, the differences were not significant, P>0.05. After 6 months of treatment, the composition ratios of the number of people in each risk level in each group were (26 / 10 / 25 in the treatment group, 36 / 12 / 12 in the control group). After 12 months of treatment, the composition ratios of the number of people in each risk level in each group were (22 / 7 / 32 in the treatment group, 34 / 8 / 18 in the control group). After 6 and 12 months of treatment, compared with before treatment in the traditional Chinese medicine compound preparation for preventing liver cancer, the number of high-risk and medium-risk people decreased compared with before, and the number of low-risk people increased compared with before. After statistical comparison, the differences were significant, P<0.05. After 12 months of treatment, after statistical processing of the comparison between the traditional Chinese medicine compound preparation treatment group for preventing liver cancer and the control group, there were obvious differences, P<0.05.

[0149] 3.8 Comparison of safety between the two groups

[0150] Through the analysis of the blood routine, urine routine, stool routine, liver and kidney functions, and electrocardiograms of the two groups of cases before and after medication, it was considered that no abnormal clinical changes related to the drugs occurred in each index, indicating that the test drug and the control drug had good safety.

[0151] 4 Discussion

[0152] Liver fibrosis is the common pathological basis of all chronic liver diseases. Slowing down, preventing, and reversing the occurrence and development of this pathological process can prevent and treat liver cirrhosis and liver cancer. Essentially, the treatment of chronic liver diseases is the treatment of liver fibrosis. The world-renowned liver disease authority Hans Popper once said, "Whoever can prevent liver fibrosis can treat most chronic liver diseases." In recent years, liver disease researchers and clinicians at home and abroad have reached a consensus that there are more than a hundred traditional Chinese and Western medicines for liver protection therapy in the past. After long-term research and observation, most of them have not been proven to be effective, and may even promote liver fibrosis. Even as adjuvant drugs, they need to be re-evaluated, and most of them do not need to be used or should be used as little as possible. Traditional Chinese medicine has potential advantages in anti-liver fibrosis, but its efficacy also needs to be confirmed by experiments and clinical practice. The gap between the experimental results and clinical efficacy of many drugs is very large. The main reasons are as follows: First, viral liver fibrosis is the main type in clinical chronic liver diseases (accounting for the vast majority), and among them, chronic hepatitis B liver fibrosis is the main one. However, previous experiments lacked animal models of viral liver fibrosis, so a reasonable, objective, and scientific evaluation of such anti-liver fibrosis drugs could not be made. Second, there is a lack of highly effective and low-toxic antiviral drugs. Third, antiviral and anti-liver fibrosis have not been organically combined. Therefore, developing a highly effective and safe new drug with the synergistic pharmacological effects of both anti-HBV and anti-liver fibrosis to meet the urgent need for clinical medication has become a research hotspot at home and abroad in recent years.

[0153] Liver fibrosis is essentially a repair response to chronic liver injury. In recent years, there has been a further understanding of the formation mechanism of liver fibrosis. Some studies have shown that hepatic stellate cells (HSCs) activated by chronic liver inflammation are the main source of extracellular matrix, and can secrete various extracellular matrix components such as type I, III, and IV collagens, LN, fibronectin, and HA. The increase in PCⅢ is related to the increase in the synthesis of type III collagen, and its serum level is positively correlated with the histological fibrosis degree. The metabolism of HA mainly occurs in liver endothelial cells, and its value is the lowest in patients with mild chronic hepatitis, and shows a gradient increase in the values of acute hepatitis, moderate chronic hepatitis, severe chronic hepatitis, and liver cirrhosis due to hepatitis in turn, indicating that HA can better reflect the degree of liver fibrosis. The average value of PCⅢ is the smallest in patients with mild chronic hepatitis, followed by liver cirrhosis due to hepatitis, and then in turn by acute hepatitis, moderate chronic hepatitis, and severe chronic hepatitis. The results of this clinical observation show that the traditional Chinese medicine compound capsule for preventing liver cancer has a significant improvement effect on the serum liver fibrosis marker indexes, and also has a good effect in improving clinical symptoms, restoring liver function, and inhibiting HBV replication, suggesting that the traditional Chinese medicine compound capsule for preventing liver cancer has the therapeutic effects of antiviral (HBV) and anti-liver fibrosis (improving the microenvironment of liver regeneration).

[0154] Through a series of clinical and experimental studies, the applicant has found that the liver regeneration microenvironment is an important mechanism for the occurrence, development, recurrence, and metastasis of liver cancer. During the progression of CHB, the deteriorating microenvironment of abnormal liver regeneration is a necessary condition and a key factor for initiating and promoting the occurrence, development, and metastasis of liver cancer. Maintaining or promoting the normal liver regeneration microenvironment and avoiding or improving the deteriorating microenvironment of abnormal liver regeneration in patients with chronic liver diseases are effective ways to delay, block, and reverse the progression of liver cancer. Regulating the "dynamic imbalance between normal liver regeneration and repair and abnormal liver regeneration disorder" is an important strategy for preventing and treating liver cancer by improving the liver regeneration microenvironment. Liver fibrosis is one of the main forms of abnormal liver regeneration. Preventing and treating the occurrence and development of liver cancer by improving the liver regeneration microenvironment through anti-liver fibrosis is a new prevention and treatment strategy. However, there is currently a lack of traditional Chinese medicine preparations that can reduce the risk of liver cancer by improving the liver regeneration microenvironment. This clinical study found that whether using the REACH-B liver cancer risk assessment model widely applied internationally or the LI-MA model established by us for evaluation, this traditional Chinese medicine compound capsule for preventing liver cancer can reduce the risk of liver cancer in patients with chronic hepatitis B while playing a role in anti-liver fibrosis and improving the liver regeneration microenvironment.

Claims

1. A traditional Chinese medicine compound for improving the hepatic regeneration microenvironment and reducing the risk of liver cancer occurrence, characterized in that, The traditional Chinese medicine compound is prepared from the following raw materials in parts by weight: 1-30 parts of mistletoe, 1-30 parts of dodder seed, 1-10 parts of turmeric, 1-50 parts of oldenlandia diffusa, and 1-60 parts of coix seed.

2. The traditional Chinese medicine compound according to claim 1, wherein The traditional Chinese medicine compound is prepared from the following raw materials in parts by weight: 7-30 parts of mistletoe, 5-25 parts of dodder seed, 5-10 parts of turmeric, 7-40 parts of oldenlandia diffusa, and 7-40 parts of coix seed.

3. The traditional Chinese medicine compound according to claim 1, wherein, The traditional Chinese medicine compound is prepared from the following raw materials in parts by weight: 7-15 parts of mistletoe, 5-10 parts of dodder seed, 5-10 parts of turmeric, 7-15 parts of oldenlandia diffusa, and 7-15 parts of coix seed.

4. A traditional Chinese medicine compound preparation, characterized in that, The preparation comprises the traditional Chinese medicine compound according to claim 1 or 2 or 3 and pharmaceutically acceptable excipients.

5. A preparation method of the traditional Chinese medicine compound preparation as described in claim 4, characterized in that: Take the five traditional Chinese medicine pieces of mistletoe, dodder seed, turmeric, oldenlandia diffusa, and coix seed according to parts by weight, decoct twice with water for 2 hours each time, combine the decoctions, filter, let the filtrate stand overnight, take the supernatant, concentrate it to a thick extract, and store the extract for later use; take the above thick extract, add an appropriate amount of starch, mix evenly, place it in a drying oven, and dry it at a temperature below 80°C; take out the dried product, weigh it, crush it, and then add starch to make the total weight reach 450 g, mix evenly, and set aside; take the above medicinal powder, make soft materials with 80% ethanol, granulate, and dry it at a temperature below 80°C, and size the granules; take the above granules, add 0.3% magnesium stearate, mix evenly, and fill them into capsules.

6. Use of the traditional Chinese medicine compound according to claim 1 in the preparation of a drug for improving the hepatic regeneration microenvironment and reducing the risk of liver cancer occurrence.

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