CN118806832B

CN118806832B - Traditional Chinese medicine composition for advanced primary liver cancer and application thereof

Info

Publication number: CN118806832B
Application number: CN202410809675.4A
Authority: CN
Inventors: 王�忠; 侯智永; 于亚南; 刘骏; 王永炎
Original assignee: Institute of Basic Research In Clinical Medicine of CACMS
Current assignee: Institute of Basic Research In Clinical Medicine of CACMS
Priority date: 2024-06-21
Filing date: 2024-06-21
Publication date: 2026-01-27
Anticipated expiration: 2044-06-21
Also published as: CN118806832A

Abstract

This invention discloses a traditional Chinese medicine composition for advanced primary liver cancer, as well as a drug comprising this composition and a method for its preparation. This invention also provides the application of the said traditional Chinese medicine composition and the drug in the treatment of advanced primary liver cancer, particularly in the treatment of unresectable advanced primary liver cancer. The traditional Chinese medicine composition of this invention comprises 1-50 parts of ginkgo, 1-35 parts of emblica, 1-30 parts of centipeda minima, 1-30 parts of artemisia annua, 1-35 parts of tangerine peel, 1-35 parts of immature bitter orange, 1-20 parts of tangerine peel, 1-30 parts of green tangerine peel, and 1-35 parts of immature bitter orange peel. This invention provides a new clinical treatment option for patients with advanced primary liver cancer.

Description

Traditional Chinese medicine composition for advanced primary liver cancer and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a traditional Chinese medicine composition for advanced primary liver cancer and application thereof.

Background

Primary liver cancer (Primary Carcinoma of Liver, PLC) refers to malignant cancers that occur in hepatocytes or intrahepatic cholangiocytes, with hepatocellular carcinoma accounting for the vast majority of primary liver cancers. Primary liver cancer is one of the most common and most dangerous malignant tumors. In recent years, the incidence rate of primary liver cancer has an increasing trend on the global scope, the incidence rate and the death rate of the primary liver cancer are 5 th and 3 rd (Sung H, etc. .Global Cancer Statistics 2020:GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36Cancers in 185Countries.CA Cancer J Clin.2021;71(3):209-249.doi:10.3322/caac.21660). th modern medical treatment on malignant tumors, etc.) the treatment scheme is mainly selected according to the stage of the liver cancer, and comprises various biotechnology such as operation treatment (hepatectomy, liver transplantation), ablation treatment, hepatic arterial embolism chemotherapy (TACE), molecular targeted drug treatment, antiviral treatment, biological treatment, radiation treatment, comprehensive chemotherapy, rapid development of immunity, genes, endocrine and the like, wherein the operation treatment is the most effective method, but most primary liver cancer patients have no obvious manifestation in the initial stage of the incidence, and once the disease is confirmed to be more advanced to the middle and late stages, most of the patients lose the operation opportunity at the moment, and the aim of radical treatment cannot be realized.

The primary liver cancer belongs to the categories of 'mass, accumulation, tympanites, accumulation syndrome, bufebrile disease' and the like in the theory of traditional Chinese medicine. Most doctors consider that the most important and fundamental pathogenesis is mainly deficiency and impairment and accumulation and toxin stasis, and is mainly malignant disease with deficiency, accumulation and accumulation of the principal deficiency and excess, which is formed by the accumulation of the principal deficiency and accumulation of the principal and principal aspects and accumulation of the principal aspects and the secondary aspects of the malignant disease, such as dysfunction of viscera, abnormal qi, blood and body fluid, qi stagnation, blood stasis, turbid phlegm, turbid dampness, heat toxin and the like, due to the factors of deficiency of healthy qi, pathogenic toxin, emotional depression, eating injury, old and old diseases and the like.

Aiming at the characteristics of the deficiency, the excess, the deficiency and the excess of the liver cancer patient, traditional Chinese medicine is mainly used for treating liver cancer by strengthening body resistance and eliminating evil the principle of treating both principal and secondary aspect of disease. The treatment of the symptoms usually includes dispersing stagnated liver qi, promoting blood circulation for removing blood stasis, clearing heat and promoting diuresis, purging pathogenic fire and removing toxic substances, resolving food stagnation and resolving hard mass, and the treatment of the root cause usually includes strengthening spleen and invigorating qi, nourishing blood and liver, and nourishing yin. The Chinese patent application of publication No. CN 105853895A discloses a traditional Chinese medicine composition for adjuvant therapy of middle-late stage primary liver cancer, which is prepared from the following traditional Chinese medicine raw materials, by weight, 120-200 parts of rhizoma curcumae, 100-150 parts of fritillaria thunbergii, 120-200 parts of rhizoma sparganii, 150-280 parts of herba scutellariae barbatae, 150-180 parts of herba patriniae, 120-150 parts of bighead atractylodes rhizome, 50-120 parts of coix seed, 10-50 parts of leech, 100-300 parts of astragalus mongholicus, 20-60 parts of ginseng, 100-200 parts of Chinese angelica sinensis, 100-200 parts of glossy privet fruit and 30-100 parts of liquorice. Also disclosed in the Chinese patent application of publication No. CN 107468949A is a traditional Chinese medicine composition for treating advanced primary liver cancer, which is prepared from 10-30 parts of radix codonopsitis, 30-50 parts of semen coicis, 15-30 parts of Chinese fanpalm seeds, 6-15 parts of wampee core, 6-15 parts of waxberry bark, 15-30 parts of corydalis saxicola bunting, 5-35 parts of waxberry root, 10-35 parts of carambola leaf and 3-9 parts of leech. Both the above two compositions embody the treatment rules and the formula principles of activating blood circulation, removing stasis, detoxifying, resolving masses, strengthening spleen, supplementing qi, nourishing yin and softening liver.

Although both drug research and clinical workers are devoted to research and development of drugs for treating liver cancer, the high mortality rate of primary liver cancer, especially advanced primary liver cancer, still suggests that the clinical therapeutic needs are far from being met.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a pharmaceutical composition for treating advanced primary liver cancer. The traditional Chinese medicine composition has novel prescription thought and strict composition, and preliminary experiments show that the traditional Chinese medicine composition has remarkable inhibition effect on the growth of human liver cancer cell strains (BEL-7402, MHCC-97H and SMMC-7721) and mouse liver cancer cell strains (H22 and Hepa 1-6), and can be used for treating advanced primary liver cancer. Through preliminary clinical observation, the traditional Chinese medicine composition disclosed by the invention can benefit patients with middle and late stage primary liver cancer. Therefore, the invention is expected to provide more choices for clinically treating primary liver cancer, especially non-operable advanced primary liver cancer.

In order to achieve the technical effects, the invention adopts the following technical scheme:

a traditional Chinese medicine composition for advanced primary liver cancer comprises the following raw materials in parts by weight:

1-50 parts of gingko, 1-35 parts of phyllanthus emblica, 1-30 parts of centipeda minima, 1-30 parts of sweet wormwood herb, 1-35 parts of dried orange peel, 1-35 parts of immature bitter orange, 1-20 parts of pummelo peel, 1-30 parts of green tangerine peel and 1-35 parts of bitter orange.

Preferably, the traditional Chinese medicine composition for advanced primary liver cancer comprises the following raw materials in parts by weight:

10-50 parts of gingko, 5-35 parts of phyllanthus emblica, 5-30 parts of centipeda minima, 5-30 parts of sweet wormwood herb, 5-35 parts of dried orange peel, 5-35 parts of immature bitter orange, 2-20 parts of pummelo peel, 5-30 parts of green tangerine peel and 5-35 parts of bitter orange.

Also preferably, the traditional Chinese medicine composition for advanced primary liver cancer comprises the following raw materials in parts by weight:

24-50 parts of gingko, 14-35 parts of phyllanthus emblica, 15-30 parts of centipeda minima, 15-30 parts of sweet wormwood herb, 14-35 parts of dried orange peel, 18-35 parts of immature bitter orange, 9-20 parts of pummelo peel, 5-30 parts of green tangerine peel and 5-35 parts of bitter orange.

More preferably, the traditional Chinese medicine composition for advanced primary liver cancer comprises the following raw materials in parts by weight:

32-50 parts of gingko, 22-35 parts of phyllanthus emblica, 20-30 parts of centipeda minima, 20-30 parts of sweet wormwood herb, 22-35 parts of dried orange peel, 24-35 parts of immature bitter orange, 12-20 parts of pummelo peel, 20-30 parts of green tangerine peel and 22-35 parts of bitter orange.

42-45 parts of gingko, 26-30 parts of phyllanthus emblica, 24-28 parts of centipeda minima, 21-25 parts of sweet wormwood herb, 26-30 parts of dried orange peel, 26-30 parts of immature bitter orange, 15-20 parts of pummelo peel, 21-25 parts of green tangerine peel and 26-30 parts of bitter orange.

As a most preferred embodiment, the invention provides a traditional Chinese medicine composition for advanced primary liver cancer, which comprises the following raw materials in parts by weight:

45 parts of gingko, 28 parts of phyllanthus emblica, 25 parts of centipeda minima, 25 parts of sweet wormwood herb, 28 parts of dried orange peel, 30 parts of immature bitter orange, 16 parts of pummelo peel, 25 parts of green tangerine peel and 28 parts of bitter orange.

As a preferred embodiment, the traditional Chinese medicine composition for advanced primary liver cancer provided by the invention is prepared from the following raw materials of gingko, phyllanthus emblica, centipeda minima, sweet wormwood, dried orange peel, immature bitter orange, pummelo peel, green tangerine peel and fructus aurantii, wherein the weight parts of the raw materials are defined as above.

The invention also aims at providing a medicament, which comprises the traditional Chinese medicine composition and can also comprise or not comprise pharmaceutically acceptable auxiliary materials.

As an alternative embodiment, the medicament is prepared from the traditional Chinese medicine composition and, optionally, pharmaceutically acceptable auxiliary materials.

Preferably, the medicament is any clinically acceptable formulation, preferably an oral formulation.

Preferably, the oral preparation is selected from decoction, powder, capsule, tablet, honeyed pill, water pill, concentrated pill, paste pill, wax pill, granule, oral liquid or dripping pill, and more preferably decoction, tablet, granule or capsule.

The third object of the present invention is to provide a preparation method of the above-mentioned medicines, which comprises preparing each medicine material according to the weight portions, and preparing clinically acceptable preparations according to the conventional method in the field with or without adding pharmaceutically acceptable auxiliary materials.

Pharmaceutically acceptable excipients include, but are not limited to, (1) diluents such as starch, powdered sugar, dextrin, lactose, pregelatinized starch, microcrystalline cellulose, inorganic calcium salts (e.g., calcium sulfate, dibasic calcium phosphate, medicinal calcium carbonate, etc.), mannitol, etc., vegetable oils, polyethylene glycols, etc., (2) binders such as distilled water, ethanol, starch slurry, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, hydroxypropyl methylcellulose, etc., (3) disintegrants such as dry starch, sodium carboxymethyl starch, low substituted hydroxypropyl cellulose, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethyl cellulose, etc., (4) lubricants such as magnesium stearate, micronized silica gel, talc, hydrogenated vegetable oils, polyethylene glycols, magnesium lauryl sulfate, etc., (5) solvents such as water, ethanol solutions, etc.

As an exemplary embodiment, the present invention provides a preparation method of a decoction, which comprises the steps of preparing each medicinal material of the traditional Chinese medicine composition according to the weight parts, adding water for decocting for two to three times, merging decoctions, and filtering.

The preparation method of the decoction comprises the steps of adding water for each decoction and the time for each decoction, wherein the parameters can be easily determined by a person skilled in the art according to the preparation rule of the traditional Chinese medicine decoction and the actual dosage of the medicinal materials.

The invention also provides a preparation method of the granule, which comprises the steps of preparing each medicinal material of the traditional Chinese medicine composition according to the weight parts, adding water 10 times of the total weight of the medicinal material, soaking for 1-2 hours, extracting volatile oil, collecting the volatile oil by a further device for later use, collecting distilled liquid medicine by a further device, taking beta-cyclodextrin 10 times of the volatile oil, adding an appropriate amount of water, dissolving the beta-cyclodextrin into the liquid medicine to form an aqueous solution at 50-70 ℃, cooling, adding the volatile oil, stirring for 1-2 hours at 300-500r/min, refrigerating for 24 hours, taking out, filtering, drying residues at 40 ℃ and grinding to obtain beta-cyclodextrin inclusion compound, keeping the residues for later use, adding water 8 times of the residues for each time, filtering when the residues are hot, merging the filtrate, adding the distilled liquid medicine to concentrate the extractum with the relative density of 1.20-1.25 (50 ℃), drying, crushing the extractum into fine powder, adding the beta-cyclodextrin inclusion compound, adding or not adding pharmaceutically acceptable auxiliary materials, uniformly mixing, granulating, and drying to obtain the granule.

As another exemplary embodiment, the invention also provides a preparation method of the capsule, which comprises the steps of preparing the medicinal materials of the traditional Chinese medicine composition according to the weight parts, independently crushing the medicinal materials into fine powder, mixing and encapsulating, or mixing the medicinal materials, crushing the medicinal materials into fine powder and encapsulating.

As another exemplary embodiment, the present invention provides a method for preparing a tablet, comprising preparing each medicinal material of the Chinese medicinal composition according to the present invention according to the parts by weight, mixing each medicinal material, pulverizing into fine powder, adding a proper amount of dextrin, and directly tabletting.

In addition, the invention also provides the application of the traditional Chinese medicine composition, the medicine and the medicine prepared according to the preparation method in preparing the medicine for treating the advanced primary liver cancer, wherein the symptoms are that the progress of the cancer disease is rapid, abdominal distension or abdominal pain is refused, the accumulation is distended, the body is emaciated, dizziness and tinnitus, low fever or high fever polydipsia, red and less body fluid tongue, thin and yellow fur and wiry and rapid pulse.

In particular, the invention also provides the application of the traditional Chinese medicine composition, the medicine and the medicine prepared according to the preparation method in preparing the medicine for treating the advanced primary liver cancer which can not be resected by surgery.

The traditional Chinese medicine composition, the medicine and the medicine prepared according to the preparation method can improve the life quality of patients with advanced primary liver cancer, protect liver function and prolong life cycle.

It is easy to understand that the Chinese medicinal composition, the medicament, and the medicament prepared according to the preparation method of the present invention can also be used for patients with intermediate primary liver cancer or postoperative liver cancer patients, and can also benefit patients.

In the present specification, "parts by weight" of each component means the relation of the amount of each component, and not the actual mass unit. The unit weight part may be any mass, for example, 1 part by weight may be 1g, 500g or 1kg, and may even be 15g, 30g, etc., according to actual circumstances.

The traditional Chinese medicine composition comprises the following components:

Semen Ginkgo is dry mature seed of Ginkgo biloba L (Ginkgo biloba L.), has sweet, bitter and astringent taste, and has effects of astringing lung, relieving asthma, astringing, stopping leukorrhagia, and reducing urination, and can be used for treating asthma, leukorrhagia, turbid urine, spermatorrhea, frequent urination, etc.

The fructus Phyllanthi is dry mature fruit of Phyllanthus emblica (Phyllanthus emblica L.) belonging to genus Phyllanthus of family Euphorbiaceae, and has sweet, sour, astringent and cool taste, and enters lung and stomach meridians. Has effects of clearing heat and cooling blood, resolving food stagnation and invigorating stomach, promoting fluid production and relieving cough, and is commonly used for treating blood heat and blood stasis, dyspepsia, abdominal distention, cough, laryngalgia and dry mouth.

The whole herb of Centipeda minima (CENTIPEDA MINIMA (L.) of the family Compositae, namely A.Braun e tAsters.) has pungent taste, warm nature, and lung meridian, has the effects of dispersing wind-cold, relieving stuffy nose, relieving cough and detoxication, and can be used for treating wind-cold common cold, stuffy nose, cough and asthma due to cold phlegm, sore and carbuncle.

Herba Artemisiae Annuae is dry aerial part of Artemisia annua (ARTEMISIA ANNUA L.) belonging to Compositae, has bitter and pungent taste, and cold nature, and can treat symptoms such as deficiency heat, removing bone steaming, clearing summer heat, checking malaria, and eliminating jaundice, and is commonly used for treating symptoms such as fever, night fever, fever due to yin deficiency, bone steaming fatigue heat, fever due to summer heat, malaria chill, and jaundice due to damp-heat.

Pericarpium Citri Tangerinae is dry mature pericarp of Citrus reticulata Blanco (Citrus reticulata Blanco) of Rutaceae and its cultivated variety, has bitter and pungent taste, warm nature, and has effects of regulating qi-flowing, invigorating spleen, eliminating dampness and resolving phlegm, and can be used for treating abdominal distention, anorexia, vomiting and diarrhea, cough and excessive phlegm.

Fructus Aurantii Immaturus is dried young fruit of Citrus aurantium (Citrus aurantium L.) of Rutaceae, its cultivar or Citrus sinensis (Citrus sinensis Osbeck), and has bitter, pungent and sour taste, slightly cold nature, and enters spleen and stomach meridians. Has the effects of removing qi-flowing, resolving food stagnation, eliminating phlegm and relieving stuffiness. Can be used for treating stagnation, distention and pain, diarrhea, constipation, phlegm stagnation, qi stagnation, chest pain, chest stuffiness, and viscera prolapse.

Exocarpium Citri Grandis is immature or near mature dried exocarp of Citrus grandis (Citrus grandis Tomentosa) or Citrus grandis (L.) Osbeck of Rutaceae, has pungent and bitter taste, warm nature, and lung and spleen meridian, has effects of regulating qi-flowing, relieving middle warmer, eliminating dampness and resolving phlegm, and can be used for treating cough with excessive phlegm, food stagnation, wine, vomiting and aversion to oppression.

The green tangerine peel is the peel of dried young fruit or immature fruit of Citrus reticulata Blanco (Citrus reticulata Blanco) of Rutaceae and its cultivated variety, has bitter and pungent taste, warm nature, and has the effects of soothing liver, breaking qi, removing food retention and resolving stagnation, and can be used for treating chest and hypochondrium distending pain, hernia pain, hyperplasia of mammary glands, acute mastitis, food retention and qi stagnation, and abdominal distention and pain.

Fructus Aurantii is dried immature fruit of Citrus aurantium (Citrus aurantium L.) of Rutaceae and its cultivated variety, has bitter, pungent and sour taste, slightly cold nature, and has effects of invigorating spleen and stomach channel, regulating qi-flowing, relieving middle-jiao, promoting stagnancy and relieving distention, and can be used for treating qi stagnation in chest and hypochondrium, distention and pain, dyspepsia, phlegm retention, internal stagnation, and viscera prolapse.

Among the nine medicinal materials, gingko is taken as a monarch drug, and the gingko is sweet, bitter and astringent in taste, and is 'cloud of Ben Cao gang mu', gingko is cooked to warm lung and benefit qi, relieve dyspnea and cough, reduce stool, stop white turbidity, promote digestion, reduce phlegm, disinfect and kill parasites. Can not only dispel cancer toxin to remove etiology, but also resolve phlegm. The centipeda minima is used for dispersing pathogenic factors, assisting the gingko to relieve cancer toxin, the phyllanthus emblica is sweet, sour and astringent, cool, clear heat and promote the production of body fluid, cool blood and promote blood circulation, promote digestion and stomach, the green tangerine peel is used for soothing liver and breaking qi, promoting digestion and resolving food stagnation, the dried orange peel is used for regulating qi and strengthening spleen, eliminating dampness and resolving phlegm, the immature bitter orange is used for breaking qi and resolving food retention, resolving phlegm and resolving masses, the five medicines are all ministerial medicines, the green tangerine peel and the immature bitter orange are compatible with the phyllanthus emblica, so that the capacity of removing blood stasis of the phyllanthus emblica is enhanced, the stomach and digestion are also promoted, the qi is regulated and the gingko is matched with the orange to enhance the gingko to resolve phlegm. In the late stage of liver cancer, deficiency heat grows internally to cause dry mouth and bitter taste, vexation and irritability, sweet wormwood and pungent taste, cold clearing and removing deficiency heat, cooling blood and relieving restlessness, exocarpium citri rubrum and dried orange peel assist gingko in resolving phlegm, fructus aurantii moves qi to open chest, and is used as an adjuvant drug for relieving swelling, and immature bitter orange and green tangerine peel enhance the force of regulating qi. The whole prescription takes qi-regulating herbs as main materials, and is compatible with the drugs for removing toxicity, removing blood stasis and resolving phlegm, so that the effects of promoting qi circulation, removing toxicity, resolving phlegm, resolving stasis, relieving all symptoms and eliminating pathogenic factors are achieved, and the effects of strengthening body resistance, eliminating pathogenic factors, detoxifying and resolving masses are achieved.

The traditional Chinese medicine composition for treating advanced primary liver cancer has novel formula and obvious inhibition effect on the growth of human liver cancer cell lines (BEL-7402 and MHCC-97H, SMMC-7721) transplanted tumors, so that the traditional Chinese medicine composition can be used for treating advanced primary liver cancer. Through clinical observation of 9 patients with advanced liver cancer (among them, the later 7 cases and the middle 2 cases), 3 patients survive (do not progress) and 6 patients die at the end of the observation period, and the administration days and the survival time have certain correlation, namely, the longer the administration time, the longer the survival time. And the related liver function index of the patient has a trend of improvement. Proved by the demonstration, the traditional Chinese medicine composition can benefit patients with advanced liver cancer.

Drawings

The invention is further described below with reference to the accompanying drawings.

FIG. 1 shows the change in tumor volume of each group of female tumor-bearing nude mice (humanized liver cancer MHCC-97H cells) in experimental example 2.

FIG. 2 shows the survival time curves of the groups of male tumor-bearing nude mice (human liver cancer MHCC-97H cells) in Experimental example 2.

FIG. 3 shows the change in tumor volume of each group of female tumor-bearing nude mice (SMMC-7721 cells of human liver cancer) in experimental example 2.

FIG. 4 is a graph showing the survival time of each group of female tumor-bearing nude mice (SMMC-7721 cells of human liver cancer) in Experimental example 2.

FIG. 5 is a graph showing the survival time of each group of male tumor-bearing nude mice (SMMC-7721 cells of human liver cancer) in Experimental example 2.

Fig. 6 shows a survival curve of patients per day in clinical observations.

Figure 7 shows a patient survival curve in months in a clinical observation.

Detailed Description

The invention is described below with reference to specific examples. It will be appreciated by those skilled in the art that these examples are for illustration of the invention only and are not intended to limit the scope of the invention in any way.

The experimental methods in the following examples are conventional methods unless otherwise specified. The materials, reagent materials, etc. used in the examples described below are all commercially available products unless otherwise specified.

The english abbreviations and chinese meanings that may occur herein are as follows:

ALT is glutamic pyruvic transaminase;

AST, glutamic-oxaloacetic transaminase;

TBIL, total bilirubin;

DBIL, direct bilirubin;

ALB, albumin;

AKP, alkaline phosphatase;

r-GGT glutamyl transpeptidase;

AFP, alpha fetoprotein;

BUN is urea nitrogen;

CR is creatinine.

Example 1A Chinese medicinal composition for treating advanced primary liver cancer

The raw materials of the traditional Chinese medicine composition of this embodiment are as follows (1 part=1 kg):

32 parts of gingko, 22 parts of phyllanthus emblica, 20 parts of centipeda minima, 20 parts of sweet wormwood herb, 22 parts of dried orange peel, 24 parts of immature bitter orange, 12 parts of pummelo peel, 20 parts of green tangerine peel and 22 parts of bitter orange

Example 2A Chinese medicinal composition for treating advanced primary liver cancer

24 parts of gingko, 14 parts of phyllanthus emblica, 15 parts of centipeda minima, 15 parts of sweet wormwood herb, 14 parts of dried orange peel, 18 parts of immature bitter orange, 9 parts of pummelo peel, 15 parts of green tangerine peel and 14 parts of bitter orange

Example 3A Chinese medicinal composition for treating advanced primary liver cancer

Example 4A Capsule

Mixing the Chinese medicinal composition of example 1, grinding into fine powder, sieving with 100 mesh sieve, and encapsulating to obtain 0.4g crude drug/granule.

The dosage is taken three times a day, 2-5 granules are taken each time, and warm water is taken each time.

Example 5A granule (Jiuwei Zhuxiao granule)

The prescription of the traditional Chinese medicine composition for preparing the granules in the embodiment is (12 g=1 part by weight):

523g of gingko, 349g of phyllanthus emblica, 314g of centipeda minima, 279g of sweet wormwood, 349g of dried orange peel, 349g of immature bitter orange, 209g of pummelo peel, 279g of green tangerine peel and 349g of bitter orange.

The preparation method comprises the steps of adding water 10 times of the total weight of the medicinal materials into nine materials, soaking for 1-2 hours, extracting volatile oil, collecting the volatile oil in a container for later use, collecting distilled liquid medicine in a container, taking 10 times of the weight of the volatile oil of beta-cyclodextrin, adding a proper amount of water, dissolving the volatile oil into the liquid medicine at 50-70 ℃, cooling the liquid medicine, adding the volatile oil into the liquid medicine, stirring the liquid medicine for 1-2 hours at 300-500r/min, refrigerating the liquid medicine for 24 hours, taking out the liquid medicine, filtering the liquid medicine, drying the residue at 40 ℃ for grinding the residue to obtain a beta-cyclodextrin inclusion compound, continuously decocting the residue with water twice, adding 8 times of water each time, decocting the residue for 1-2 hours, filtering the liquid medicine while the liquid medicine is hot, merging the filtrate, adding the distilled liquid medicine into extractum with the relative density of 1.20-1.25 (50 ℃) for drying the liquid medicine, crushing the liquid medicine into fine powder, adding the beta-cyclodextrin inclusion compound, adding pharmaceutically acceptable auxiliary materials into the liquid medicine, uniformly mixing, granulating the mixture, drying the mixture, granulating, and granulating the mixture, and obtaining 1000g, and subpackaging each bag.

The dosage is taken three times a day, 7g each time, and with boiled water.

Example 6A tablet

Mixing the Chinese medicinal composition of example 3, grinding into fine powder, sieving with 80-100 mesh sieve, adding dextrin, and directly pressing into tablet, and making into tablet with 0.3g crude drug/tablet.

The dosage is 3-6 tablets each time, three times daily, and taken with warm water.

EXAMPLE 7A honeyed pill

Mixing the Chinese medicinal composition of example 1, grinding into fine powder, sieving with 60-100 mesh sieve, adding appropriate refined honey, and making into small honeyed pill, 3g/10 pill by conventional method.

The dosage is taken three times a day with 3g each time with warm water.

Example 8A decoction

Weighing the following medicinal materials by mass:

15g of gingko, 10g of phyllanthus emblica, 9g of centipeda minima, 8g of sweet wormwood, 10g of dried orange peel, 10g of immature bitter orange, 6g of pummelo peel, 8g of green tangerine peel and 10g of bitter orange

Mixing the above materials, adding 5-10 times of water, boiling, decocting for 25 min, hot filtering, adding 5-10 times of water into the residue, heating to boil, decocting for 20-30 min, hot filtering, and mixing the two decoctions. The medicine is taken for 2-3 times in 1 day.

Experimental example 1 in vitro pharmacological experiment

1. Material

1.1 Experimental drugs:

① The Jiuwei Zhaoxiao granule is prepared according to the prescription and the method of the example 5, the batch number is 20200101, the content is 3.70g crude drug/g extract powder, the Jiuwei Zhaoxiao granule is provided by clinical basic research institute of Chinese medical science, and is received at 21 days of 01 month in 2020, and is stored in a cool cabinet in a drying way.

② Ganfule Capsule, batch number 20190817, kangpu pharmaceutical Co., ltd

Radix Codonopsis, carapax Trionycis (processed with vinegar), rhizoma paridis, atractylodis rhizoma (parched), radix astragali, pericarpium Citri Tangerinae, eupolyphaga Seu Steleophaga, radix et rhizoma Rhei, semen Persicae, herba Scutellariae Barbatae, herba Patriniae, poria, coicis semen, radix Curcumae, lignum sappan, concha Ostreae, herba Artemisiae Scopariae, caulis Akebiae, rhizoma Cyperi (processed), lignum Aquilariae Resinatum, and bupleuri radix.

[ Specification ] 0.5 g/grain.

1.2 Experimental reagents and cell lines:

DMEM high-sugar culture medium with the specification of 500 mL/bottle, batch No. AF29498408, hyClone company, RPMI-1640 culture medium with the specification of 500 mL/bottle, batch No. AF29498408, hyClone company, fetal Bovine Serum (FBS) with the specification of 100 mL/bottle, batch No. M009-6, scientific company.

Human hepatoma cells (BEL-7402), human hepatoma cells (SMMC-7721, low differentiation, HBV positive), mouse hepatoma cells (H22, ascites type), mouse hepatoma cells (Hepa 1-6) all from the Living technologies of GmbH, and human hepatoma cells (MHCC-97H, low differentiation, high metastasis) from Shanghai Fuxiang Biotechnology.

BEL-7402 cells, H22 cells, SMMC-7721 cells were cultured in RPMI-1640,10% fetal bovine serum at 37deg.C in a saturated humidity environment. The culture conditions of the Hepa 1-6 cells and the MHCC-97H cells are that the cell culture medium is DMEM,10% fetal calf serum, 37 ℃ and 5% CO2, and the culture is carried out in a saturated humidity environment.

1.3 Laboratory apparatus:

the medical purification workbench of the CJ-1F type comprises a center number 176, a CO2 incubator of model 3111, a center number 024, thermo Fisher company of America, a DMIL type inverted fluorescence microscope, a center number 027, leica company of Germany, a Spectra Max i3x type multifunctional enzyme labeling instrument, a center number 525 and an MD company of America.

2. Method of

2.1 Grouping and concentration setting:

According to the solubility of the nine-ingredient mass-eliminating particles and the Ganfule capsules, 6 concentration gradients of 10000, 3000, 1000, 300, 100 and 30 mug (particle or capsule content)/mL are sequentially arranged, the preparation concentrations of the nine-ingredient mass-eliminating particles and the Ganfule capsules are 20000 (maximum soluble concentration), 6000, 2000, 600, 200 and 60 mug/mL respectively, a solvent control group is additionally arranged, 5 compound holes are arranged at each concentration, 4 zeroing holes without adding cells are additionally arranged at each concentration, and the test is repeated for three batches. The concentrations of the test substances in the second and third tests were adjusted to 1000, 300, 100, 30, 10 and 3. Mu.g/mL according to the results of the first test. The details of the group administration are shown in Table 1.

Table 1 group administration table

2.2 Cell culture:

BEL-7402, H22, SMMC-7721 cell suspensions were seeded into 96-well plates at 100. Mu.L/well, 200. Mu.L/well PBS was added, 2 96-well plates were seeded per cell, and cultured overnight until cells attached. The cell seeding sequence of the 96-well plate is shown in Table 2.

Table 2 sample loading diagram for 96-well plate

1

2

3

4

5

6

7

8

9

10

11

12

A

PBS

B

PBS

Cells

Blank space

PBS

C

PBS

Cells

Blank space

PBS

D

PBS

Cells

Blank space

PBS

E

PBS

Cells

Blank space

PBS

F

PBS

Cells

Blank space

PBS

G

PBS

Cells

Blank space

PBS

H

PBS

2.3 Sample/control treatment:

Nine kinds of mass eliminating granule or Ganfule capsule in different concentration are prepared, 100. Mu.L/hole is added into 96 Kong Bankong prepared in step 2.2, and test article/reference article is added into cell inoculated culture hole or blank hole (3-11 columns, 9 holes in total) in the arrangement mode of table 2, and 100. Mu.L/hole complete culture medium is added into the 2 nd column hole. Final concentrations were 10000, 3000, 1000, 300, 100, 30 μg/mL wells/concentration, 5 replicates were made per group, 4 zeroing wells with no cells added to the sample. And (3) performing dry prognosis by adopting liver cancer cells corresponding to a test sample or a control sample, and placing a 96-well plate in a saturated humidity environment with CO2 at 37 ℃ for 72 hours. Three batches were repeated for each cell assay, and the second and third batch concentrations were adjusted to 1000, 300, 100, 30, 10, 3 μg/mL based on the results of the first batch.

2.4 Cell viability assay:

After 72h of cell culture, 20. Mu.L of MTT solution (5 mg/mL) was added to each well, and the culture was continued for 4h. After the culture was terminated, the culture solution in the wells was aspirated, 150. Mu.L of dimethyl sulfoxide (DMSO) was added to dissolve crystals, absorbance values of each well at OD492nm were detected by a multifunctional microplate reader, and cell growth inhibition ratio (%) = (OD vehicle-OD experimental group)/(OD vehicle-OD background) ×100% was calculated, and cell half-maximal growth inhibition concentration (IC 50) was calculated.

2.5 Statistical method:

The effective number of the test data was modified by rounding, the cell inhibition rate was expressed as a percentage, and the IC50 of half-effective inhibition concentration was counted using SPSS 19.0.

3. Experimental results

3.1 First test results:

As shown in Table 3, the ICs ₅₀ of the nine-ingredient mass-eliminating particles on human liver cancer BEL-7402 cells, MHCC-97H cells, SMMC-7721 cells, and murine H22 cells and Hepa1-6 cells were 517.919, 16.782, 782.321, <30.000, 161.348. Mu.g/mL, respectively, and 95% confidence intervals could not be calculated. IC ₅₀ of Ganfule capsules on human liver cancer BEL-7402 cells, MHCC-97H cells, SMMC-7721 cells, murine H22 cells and Hepa1-6 cells are 72.919, 17.720, 396.233, <30.000 and 3.201 mug/mL respectively, wherein 95% confidence intervals on human liver cancer BEL-7402 cells, MHCC-97H cells and SMMC-7721 cells are 0.835-261.626, 0.118-55.727 and 45.824-2531.187 mug/mL respectively. According to the results, the effective concentrations of the nine-ingredient mass-eliminating particles and liver-complex capsule in-vitro anti-liver cancer cells are obviously less than 10000 and 3000 mug/mL, and the concentrations of the tested substances are adjusted to 1000, 300, 100, 30, 10 and 3 mug/mL in the second and third experiments in order to obtain the IC50 value more accurately.

TABLE 3 influence of Jiuwei Zhuxiao particles on proliferation of different liver cancer cells (first batch)

3.2 Second and third test results the results are shown in tables 4 and 5.

The data in tables 4 and 5 show that nine types of mass-eliminating particles have IC ₅₀ of 243.429 μg/mL and 24.397 μg/mL for BEL-7402 cells, IC ₅₀ of 0.151 μg/mL and <3 μg/mL for H22 cells, IC ₅₀ of 8.772 μg/mL and 2.590 μg/mL for Hepa 1-6 cells, IC ₅₀ of 0.971 μg/mL and 1.841 μg/mL for MHCC-97H cells, and IC ₅₀ of >4.922 μg/mL and 116.252 μg/mL for SMMC-7721 cells, respectively.

The Ganfule capsules had IC ₅₀ for BEL-7402 cells of 600.121. Mu.g/mL and 65.660. Mu.g/mL, respectively, IC ₅₀ for H22 cells of 0.001. Mu.g/mL and 0.018. Mu.g/mL, IC ₅₀ for Hepa1-6 cells of 2.950. Mu.g/mL and 7.729. Mu.g/mL, IC ₅₀ for MHCC-97H cells of 0.041. Mu.g/mL and 0.229. Mu.g/mL, respectively, and IC ₅₀ for SMMC-7721 cells of 21215.318. Mu.g/mL and 2659.064. Mu.g/mL, respectively.

4. Conclusion of experiment:

The results show that the nine-ingredient mass-eliminating particles and the Ganfule capsules have obvious inhibition effects on in vitro proliferation of liver cancer cells (BEL-7402, H22, hepa 1-6 and MHCC-97H, SMMC-7721) from different sources, and the in vitro inhibition effect of the nine-ingredient mass-eliminating particles at 3-1000 mug/mL is superior to that of the Ganfule capsules.

TABLE 4 influence of Jiuwei Zhuxiao particles on proliferation of different liver cancer cells (second batch)

TABLE 5 influence of Jiuwei Zhuxiao particles on proliferation of different liver cancer cells (third batch)

Experimental example 2 in vivo pharmacodynamics study

Human liver cancer SMMC-7721 cells (low differentiation, HBV positive) and MHCC-97H cells (low differentiation, high metastasis) are selected for constructing a BALB/c nude mice subcutaneous transplantation tumor model, and the in vivo anti-tumor drug effect of the traditional Chinese medicine composition is evaluated by observing indexes such as tumor volume, survival time and the like.

1. Experimental materials

1.1 Test article

Nine kinds of granule for eliminating mass are prepared according to the method of example 5, wherein the batch number is 20200101, and the content is 3.70g crude drug/g extract powder.

1.2 Positive control

Ganfule capsules with the specification of 0.5 g/granule, batch number of 20190817/20201006, effective period of 2021, 08, 12, 2022 and 10, and production unit of Kangpu pharmaceutical Co Ltd.

1.3 Laboratory animals

The SPF-grade BALB/c nude mice are passaged in vivo by 20, the male and female are respectively half, the animal quality qualification number is No.1107272011004623 and No.430727201101206635, the SPF-grade BALB/c nude mice are respectively half, the animal quality qualification number is No.430727211100012884, and the SPF-grade BALB/c nude mice are purchased from Hunan Slike Lakkera laboratory animal Limited company, and the laboratory animal production license number is SCXK (Xiang) 2019-0004. The animal model is fed in a region A of a barrier environment animal laboratory of Hunan Primama drug research center, and the use license number of experimental animals is SYXK (Hunan) 2015-0016, which is used for the influence test of a human liver cancer MHCC-97H cell subcutaneous transplantation tumor model.

The SPF-grade BALB/c nude mice are passaged in vivo by 20, the number of male and female nude mice is 430727221100833945, the number of animal quality qualification certificate is 430727221100833945, the number of female and male nude mice is 120, the number of animal quality qualification certificate is 430727221101058843, the nude mice are purchased from Hunan Stokes Lekkera laboratory animal Limited, and the laboratory animal production license number is SCXK (Hunan) 2019-0004. The animals are bred in the A area of a barrier environment animal laboratory of the limited company of the Hunan Primax pharmaceutical research center, and the use license number of the experimental animals is SYXK (Hunan) 2020-0015, which is used for the influence test of the human liver cancer SMMC-7721 cell subcutaneous transplantation tumor model.

1.4 Cell lines

Human hepatoma cells (SMMC-7721, low differentiation, HBV positive) were purchased from the Living technologies Co., ltd. Of Withannoman, and human hepatoma cells (MHCC-97H, low differentiation, high metastasis) were purchased from Shanghai Fuxiang Biotechnology Co., ltd.

1.5 Major reagents

DMEM high-sugar medium (lot number: 081012-UY, product of Corning company), fetal bovine serum (lot number: NYL1009, product of HyClone company).

1.6 Main instruments

3111 Type CO ₂ incubator (Thermo in the United states), SHHW21.420A type three-purpose electric constant temperature water bath (Yongxing instruments Co., ltd. In Beijing), DMIL inverted microscope (Leica in Germany), LDZX-50KBS type vertical pressure sterilizer (Shanghai Shen An medical instruments Co., ltd.), TD4 type desk-top low-speed centrifuge (Kada industries development Co., hunan province), PL1501-S type electronic balance (Metrele Tolye instruments Co., shanghai), digital vernier caliper (Yongkang Zhengfeng hardware Co., ltd.).

2. Experimental method

2.1 Recovery culture of tumor cells

SMMC-7721 human liver cancer cell strain or MHCC-97H human liver cancer cell strain is placed in DMEM high sugar culture medium containing 10% fetal calf serum, placed in a 37 ℃ and 5% CO ₂ incubator for culture and passage for standby.

2.2 In vivo passage of tumor cells

SPF-grade BALB/c nude mice are respectively half male and female, 5.54X10 ⁷/female of human liver cancer MHCC-97H cells are subcutaneously injected into the right armpit, in vivo passage is carried out, tumor-bearing mice which are more than 2 generations are selected, tumor homogenates are prepared for standby after tumor stripping, SPF-grade BALB/c nude mice are respectively half male and female, 5.4X10 ⁷ of human liver cancer SMMC-7721 cells are subcutaneously injected into the right armpit, in vivo passage is carried out, tumor-bearing mice which are more than 2 generations are selected, and tumor homogenates are prepared for standby after tumor stripping.

2.3 Modeling, grouping and administration

2.3.1 Effect of nine kinds of granule for eliminating mass on human liver cancer MHCC-97H cell subcutaneous transplantation tumor model

70 Female and male BALB/c nude mice with qualified SPF grade are selected, and the mice tumor homogenate (MHCC-97H cells) in 2.2 is subcutaneously injected into the right armpit of the nude mice according to 0.2 mL/nude mice so as to replicate the subcutaneous transplantation tumor model of the mice. The tumor volumes of 50 tumor models of female and male mice which are formed into tumors are respectively 156.1-2247.0 mm ³ and 146.5-10238.6 mm ³, the tumor volumes are divided into 5 groups randomly according to the tumor volumes, namely a model control group and a Ganfule capsule group (1.2 g capsule content/kg and clinical equivalent dose), nine types of tumor-eliminating particles are respectively low, medium and high dose groups (4.1, 8.2 and 16.4g crude drugs/kg which are respectively equivalent to 0.5, 1 and 2 times of the clinical equivalent dose), and 10 types of tumor-eliminating particles/group/sex are selected. The nine ingredients of the dry extract powder of the granule for eliminating mass and the Ganfule capsule are prepared into corresponding concentrations by adopting pure water before daily administration. The corresponding concentration of the liquid medicine is given to each group according to the oral gavage of 20mL/kg, the equal volume of pure water is given to the model control group through the oral gavage, and the administration is carried out for 1 time/day for 4 weeks continuously. The time to death was observed for each group of mice and the number of days to live was calculated. The major (a) and minor (b) diameters of the tumor mass were measured weekly using vernier calipers and the tumor mass volume V (mm ³)＝1/2×a×b²) was calculated 2 times per week.

2.3.2 Effect of nine kinds of granule for eliminating mass on model of human liver cancer SMMC-7721 cell subcutaneous transplantation tumor

70 Female and male BALB/c nude mice with qualified SPF grade are selected, and the mouse tumor homogenate (SMMC-7721 cells) in 2.2 is inoculated under the right armpit of the nude mice according to 0.2 mL/nude mice so as to replicate the mouse subcutaneous transplantation tumor model. The tumor volumes of 50 tumor models of female and male mice which are formed into tumors are 1138.23-4669.2 mm ³ and 1378.62-3786.32 mm ³ respectively, the tumor volumes are divided into 5 groups randomly according to the layering, and the tumor volumes are respectively a model control group, a Ganfule capsule group (1.2 g capsule content/kg), nine groups of low, medium and high dosage (4.1, 8.2 and 16.4g crude drugs/kg) and 10 groups/sex. Before daily administration, pure water is adopted to prepare the nine-ingredient dry extract powder of the granule for eliminating mass and the Ganfule capsule into liquid medicine with corresponding concentration. The corresponding concentration of the liquid medicine is given to each group according to the oral gavage of 20mL/kg, the equal volume of pure water is given to the model control group through the oral gavage, and the administration is carried out for 1 time/day for 4 weeks continuously. The time to death was observed for each group of mice and the number of days to live was calculated. The major (a) and minor (b) diameters of the tumor mass were measured weekly using vernier calipers and the tumor mass volume V (mm ³)＝1/2×a×b²) was calculated 2 times per week.

2.5 Statistical methods

The significant numerical modification of this test data was performed by rounding and statistical analysis according to the SOP specifications. The software used for statistics was SPSS22.0. The measurement data is expressed as mean ± standard deviationThe normal and variance alignment were examined using Leven's test method. If there is no statistical significance (P > 0.05), statistical analysis is performed using one-way analysis of variance (ANOVA). If ANOVA is statistically significant (P≤0.05), a comparison analysis is performed using LSD test (parametric method). If the variance is not uniform (P≤0.05), then the Kruskal-Wallis test is used. If the Kruskal-Wallis Test is statistically significant (P≤0.05), a comparative analysis is performed using Dunnett's Test (nonparametric method). The statistical result takes alpha=0.05 as a checking limit, wherein p≤0.05 represents a statistical significance and p≤0.01 represents a very significant significance of the checked differences.

3. Experimental results

3.1 Effect on tumor volume of human liver cancer MHCC-97H cell subcutaneous transplantation tumor model 3.1.1 Effect on tumor volume of female mouse subcutaneous transplantation tumor model

The results are shown in Table 6 and FIG. 1. Compared with a model control group, the tumor volumes of D17, D21, D24 and D27 after the liver complex capsule group is dosed are obviously reduced (P is less than or equal to 0.05 or P is less than or equal to 0.01), the tumor volume of D27 after the nine mass-eliminating particle low-dose group is obviously reduced (P is less than or equal to 0.01), the tumor volumes of D21, D24 and D27 after the medium-dose group is dosed are obviously reduced (P is less than or equal to 0.05 or P is less than or equal to 0.01), and the tumor volumes of D17, D21, D24 and D27 after the high-dose group is dosed are obviously reduced (P is less than or equal to 0.05 or P is less than or equal to 0.01).

TABLE 6 influence of Jiuwei Zhaoxiao particles on tumor volume of female mouse subcutaneous tumor modeln=10)

Note that ⁺P≤0.05,⁺⁺ P was less than or equal to 0.01 compared to the model control group.

3.1.2 Effect on tumor volume of Male mice subcutaneously transplanted tumor model

The results are shown in Table 7, and the model group male animals were not involved in tumor volume statistics because of the subsequent death after administration, resulting in a smaller number of animals, but the animal death was counted, as described in detail in 3.2.

TABLE 7 influence of nine types of mass-eliminating particles on tumor volume of Male animals

3.2 Effect on survival time of human liver cancer MHCC-97H cell subcutaneous transplantation tumor model

The results are shown in Table 8 and FIG. 2, compared with the model control group, the survival time of the animals in the nine animals with the mass-eliminating particles and the high dose group is obviously increased (P is less than or equal to 0.05 or P is less than or equal to 0.01), the survival time of the animals in the high dose group is obviously prolonged (P is less than or equal to 0.01), the survival time of the animals in the Ganfule capsule group and the nine animals with the mass-eliminating particles and the low dose group has an increasing trend but no statistical difference, and in addition, the death condition of the female mice is not seen during the administration period.

TABLE 8 influence of nine types of mass-eliminating particles on survival time and mortality of Male transplantation tumor model

3.3 Effect on tumor volume of human liver cancer SMMC-7721 cell subcutaneous transplantation tumor model 3.3.1 Effect on tumor volume of female mouse

The results are shown in Table 9 and FIG. 3, and compared with the model control group, the tumor volume of D14 in the nine-ingredient mass-eliminating granule after the dosage group is dosed is obviously reduced (P is less than or equal to 0.05), and the tumor volumes of the low-dosage group and the high-dosage group have a tendency of reduction, but no obvious statistical difference.

TABLE 9 influence of nine types of mass-eliminating particles on tumor volume in females

Note that ⁺ P was less than or equal to 0.05 compared to the model control group, "/" indicated that the animals had all died, and no data was available.

3.3.2 Effect on tumor volume in Male mice

The results are shown in Table 10 and FIG. 4, and compared with the model control group, the liver complex capsule group and the nine-ingredient mass-eliminating granule group have no obvious difference in tumor volume of male animals.

TABLE 10 influence of nine types of mass-eliminating particles on tumor volume of male animals

3.4 Effect on survival days of human liver cancer SMMC-7721 cell subcutaneous transplantation tumor model 3.4.1 Effect on female survival period

The results are shown in Table 11 and FIG. 4, and compared with the model control group, the survival time of the nine animals of the mass-eliminating particle high-dose group female animals is obviously prolonged (P is less than or equal to 0.01).

TABLE 11 influence of nine particles on the number of days of survival of females

Note that ⁺⁺ P was less than or equal to 0.01 compared to the model control group.

3.4.2 Effect on Male animal Life cycle

The results are shown in Table 12 and FIG. 5, and compared with the model control group, nine animals with low, medium and high dosage groups of the particles for eliminating mass have a trend of prolonging the survival time of the male animals, but no statistical difference.

TABLE 12 influence of nine types of mass-eliminating particles on the number of days of survival of Male animals

4. Experimental nodule

The results show that the nine-ingredient mass-eliminating particles can obviously inhibit the growth of tumors of MHCC-97H type human liver cancer cell transplantation tumor-bearing female mice at low, medium and high doses (4.1, 8.2 and 16.4g crude drugs/kg), can obviously reduce the death rate of advanced liver cancer tumor-bearing male mice and prolong the life cycle, and the nine-ingredient mass-eliminating particles can obviously inhibit the growth of tumors of SMMC-7721 type human liver cancer cell transplantation tumor-bearing female mice at the medium dose (8.2 g crude drugs/kg) and can obviously prolong the life cycle of tumor-bearing female mice at the high dose (16.4 g crude drugs/kg). The nine-ingredient mass-eliminating particles are indicated to have obvious inhibition effect on tumor of MHCC-97H, SMMC-7721 type human liver cancer cell transplantation tumor-bearing mice.

Clinical observations the granule of the present invention reports clinical observations

The clinical curative effects of the nine-ingredient mass-eliminating granule prepared in example 5 for treating advanced liver cancer are observed in hospitals in Hubei province in 4 months 2019 to 6 months 2020.

1. Patient base conditions are shown in Table 13.

TABLE 13 clinical observation of case base conditions

2. Dosing regimen

The nine ingredients of the granule (prepared in example 5) are taken 3 times daily, 7g each time, with boiled water.

3. Evaluation of efficacy

3.1 Overall efficacy

The administration time is 3-90 days, the survival time of patients is 8-263 days, and the survival curves according to days and months are shown in fig. 6 and 7 respectively.

Fig. 6 and 7 show that the longer the administration time, the longer the survival time (administration time is 3-90 days, survival time is 8-263 days), and the correlation coefficient is 0.5669.

3.2 Physiological and biochemical index changes before and after treatment are shown in Table 14.

Table 14 changes in physiological and biochemical indicators before and after treatment

Table 14 shows that nine types of mass-eliminating particles have a tendency to decrease AFP, r-GGT, ALT, AST, etc.

In summary, the invention provides a traditional Chinese medicine composition which can be used for advanced primary liver cancer. The composition comprises gingko, phyllanthus emblica, centipeda minima, sweet wormwood, dried orange peel, immature bitter orange, exocarpium citri rubrum, green tangerine orange peel and fructus aurantii, and has the effects of regulating qi, resolving phlegm, removing toxin and removing blood stasis. Pharmacological experiments prove that the traditional Chinese medicine composition has remarkable inhibition effect on the growth of human liver cancer cell strains (BEL-7402, MHCC-97H, SMMC-7721) and mouse liver cancer cell strains (H22, hepa 1-6). Clinical observations show that the traditional Chinese medicine composition provided by the invention can benefit patients. Therefore, the invention provides a new clinical treatment choice for patients with advanced primary liver cancer and doctors.

Claims

1. A traditional Chinese medicine composition for advanced primary liver cancer is prepared from the following raw materials in parts by weight:

2. The traditional Chinese medicine composition according to claim 1, wherein the traditional Chinese medicine composition comprises the following medicinal materials in parts by weight:

3. The traditional Chinese medicine composition according to claim 2, wherein the traditional Chinese medicine composition comprises the following medicinal materials in parts by weight:

4. A medicament prepared from the traditional Chinese medicine composition of any one of claims 1 to 3 and pharmaceutically acceptable auxiliary materials.

5. The medicament according to claim 4, wherein the medicament is any one of the clinically acceptable formulations.

6. The medicament of claim 5, wherein the formulation is an oral formulation.

7. The medicament according to claim 6, wherein the oral preparation is selected from the group consisting of decoction, powder, capsule, tablet, honeyed pill, water pill, concentrated pill, paste pill, wax pill, granule, oral liquid and drop pill.

8. The medicament of claim 7, wherein the oral formulation is a decoction, a tablet, a granule, or a capsule.

9. The preparation method of the medicament as claimed in any one of claims 4 to 8, which comprises preparing each medicinal material of the traditional Chinese medicine composition according to any one of claims 1 to 3 by weight, adding pharmaceutically acceptable auxiliary materials according to a conventional method in the field, and preparing a clinically acceptable preparation.

10. A preparation method of a decoction, comprising preparing each medicinal material of the traditional Chinese medicine composition according to any one of claims 1 to 3 by weight, adding water for decoction for two to three times, combining the decoctions, and filtering.

11. A method for preparing the granule comprises the steps of preparing each medicinal material of the traditional Chinese medicine composition according to any one of claims 1-3, adding water 10 times of the total weight of the medicinal material, soaking for 1-2 hours, extracting volatile oil, collecting the volatile oil in a further device for later use, collecting distilled liquid medicine in a further device, taking beta-cyclodextrin 10 times of the volatile oil, adding a proper amount of water, dissolving the beta-cyclodextrin 50-70 ℃ into water solution, cooling, adding the volatile oil, stirring for 1-2 hours at 300-500r/min, refrigerating for 24 hours, taking out, filtering, drying residues at 40 ℃ and grinding to obtain beta-cyclodextrin inclusion compound, decocting the residues with water for later use, adding water 8 times of water each time, decocting for 1-2 hours each time, filtering while hot, merging filtrate, adding distilled liquid medicine, concentrating to an extract with the relative density of 1.20-1.25 at 50 ℃, drying, crushing into fine powder, adding the beta-cyclodextrin inclusion compound, adding or not adding pharmaceutically acceptable auxiliary materials, mixing uniformly, granulating by a dry method, and drying to obtain granules.

12. The use of the Chinese medicinal composition according to any one of claims 1 to 3, the medicament according to any one of claims 4 to 8, and the medicament prepared according to the preparation method of any one of claims 9 to 11 for preparing a medicament for treating advanced primary liver cancer, wherein the symptoms are hypochondriac pain, hypochondriac stuffiness and lump, anorexia, listlessness, dizziness, dry mouth, bitter taste, vexation and irritability, red tongue, yellow tongue coating and wiry pulse.

13. Use of the traditional Chinese medicine composition according to any one of claims 1 to 3, the medicament according to any one of claims 4 to 8, the medicament prepared according to the preparation method according to any one of claims 9 to 11, for the preparation of a medicament for the treatment of advanced primary liver cancer which cannot be resected surgically.