Application of traditional Chinese medicine composition in preparation of medicine for treating primary biliary cholangitis

Abstract

The invention relates to an application of a traditional Chinese medicine composition in preparation of a medicine for treating primary biliary cholangitis, which is characterized in that the traditional Chinese medicine composition is prepared from monkshood, dried ginger, liquorice, honeysuckle, spina gleditsiae, humulus scandens, pogostemon cablin and pericarpium citri reticulatae. The composition can obviously inhibit liver injury of a mouse primary biliary cholangitis model induced by operation and chemical induction, inhibits gene expression levels of inflammatory cytokines IL-33, chemotactic factors MCP-1 and CXCL1 in liver tissues, and has a good treatment effect on primary biliary cholangitis.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, relates to a new use of a traditional Chinese medicine composition, and specifically relates to an application of the traditional Chinese medicine composition in preparing a medicine for primary biliary cholangitis. Background Art

[0002] Primary Sclerosing Cholangitis (PSC) is a chronic, progressive autoimmune biliary disease that is usually characterized by chronic inflammation and fibrosis of the bile duct wall. This disease mainly affects the intrahepatic and extrahepatic bile ducts, which may lead to bile duct stenosis, occlusion, and cholestasis, and may eventually cause serious complications such as liver failure, cirrhosis, and biliary cancer.

[0003] The exact cause of primary biliary cholangitis is still unclear, but autoimmune reactions are thought to play a key role in its pathogenesis. The immune system mistakenly attacks the bile ducts inside and outside the liver, leading to chronic inflammation and fibrosis. This inflammation and fibrosis eventually lead to stricture and occlusion of the bile ducts, preventing bile from being discharged normally, thereby impairing liver function. Summary of the invention

[0004] According to one aspect of the present invention, there is provided a use of a traditional Chinese medicine composition in the preparation of a drug for primary biliary cholangitis, comprising the following components in parts by mass: 10-30 parts of aconite root, 5-15 parts of dried ginger, 20-30 parts of liquorice, 10-20 parts of honeysuckle, 10-30 parts of Gleditsia sinensis thorns, 20-45 parts of Ipomoea australis, 10-15 parts of patchouli, and 5-15 parts of tangerine peel.

[0005] In the prescription, aconite, dried ginger and liquorice are the main drugs, honeysuckle, honeysuckle thorn and five-clawed dragon are the assistant drugs, patchouli is the adjuvant drug, and tangerine peel is the guiding drug. Specifically:

[0006] Aconite, dried ginger, and a large amount of licorice are the main drugs, warming the lower abdomen. Wuzhaolong, also known as Nanqi, nourishes the middle and replenishes qi, and is the minister drug. Together with the three main drugs, they support the positive energy; a large amount of licorice and Wuzhaolong strengthen the spleen and replenish qi, and are used together with the adjuvant drugs Patchouli and Tangerine Peel to aromaticize the evil of dampness and turbidity; honeysuckle clears heat and detoxifies, and expel evil. Wu Jutong "treats the upper Jiao like feathers, and it will not rise unless it is light", so only 10-20 parts of honeysuckle are used; honeysuckle and Gleditsia sinensis are both minister drugs. Wuzhaolong (Nanqi) and Gleditsia sinensis are the main components of Tounong Powder. Gleditsia sinensis and honeysuckle are used together to increase the function of clearing away epidemic toxins and draining pus.

[0007] In some embodiments, the following components are included by mass: 10-20 parts of aconite, 10-15 parts of dried ginger, 20-25 parts of licorice, 10-15 parts of honeysuckle, 10-20 parts of Gleditsia sinensis, 20-30 parts of Ipomoea australis, 10-12 parts of patchouli, and 5-10 parts of tangerine peel.

[0008] In some embodiments, the following components are included in parts by mass: 10-12 parts of aconite, 12-15 parts of dried ginger, 20-22 parts of licorice, 10-12 parts of honeysuckle, 10-12 parts of Gleditsia sinensis, 20-22 parts of Ipomoea australis, 10-12 parts of patchouli, and 5-6 parts of tangerine peel.

[0009] In some embodiments, the following components are included by mass: 10 parts of aconite, 15 parts of dried ginger, 20 parts of licorice, 10 parts of honeysuckle, 10 parts of Gleditsia sinensis, 20 parts of Ipomoea australis, 10 parts of patchouli, and 5 parts of tangerine peel.

[0010] In some embodiments, the drug further comprises pharmaceutically acceptable adjuvants or additives. Specifically, the pharmaceutically acceptable adjuvants or additives include fillers, disintegrants, lubricants, suspending agents, adhesives, sweeteners, flavoring agents, preservatives, matrices, including but not limited to starch, lactose, glucose, sodium carboxymethyl cellulose, ethyl cellulose and methyl cellulose, malt, gelatin, polyols (such as propylene glycol, glycerol, mannitol), compressed tablets, etc., to help improve the stability of the pharmaceutical composition or increase the activity, or produce an acceptable taste or smell in the case of oral administration.

[0011] In some embodiments, the aforementioned composition can be prepared into granules, tablets, capsules, pills, oral solutions, oral suspensions or oral emulsions using existing traditional Chinese medicine preparation methods, thereby facilitating clinical administration.

[0012] The preparation method of the traditional Chinese medicine composition comprises the following steps: taking aconite root, dried ginger, liquorice, honeysuckle, thorn of Gleditsia sinensis, five-clawed dragon, patchouli, and tangerine peel, adding 8 to 12 times of water equivalent to the total mass of the raw materials, heating to boiling, maintaining a slight boiling for 1 to 2 hours, and obtaining a first decoction and medicinal residues after separation; a second time, taking the medicinal residues obtained after the first decoction, adding 6 to 10 times of water equivalent to the total mass of the raw materials, heating to boiling, maintaining a slight boiling for 1 to 2 hours, and obtaining a second decoction and medicinal residues after separation; combining the first decoction and the second decoction to prepare a decoction, and reducing the pressure on the decoction filtrate to concentrate to obtain a clear paste, and crushing it.

[0013] According to another aspect of the present invention, there is provided a use of any of the above compositions in the preparation of a drug for treating liver injury. The liver injury may be liver injury caused by primary biliary cholangitis.

[0014] When the present invention is used as a drug for treating primary biliary cholangitis, an oral solution can be used, and a dose can be given to severe patients once a day, divided into 2-3 oral or nasogastric administrations, once every 6-8 hours; two doses can be given to critical patients daily, divided into two oral or nasogastric administrations in the morning and evening, once every 8-12 hours; and a dose can be given to mild patients once a day, orally or nasogastric administration. Each dose is 150-250 ml.

[0015] When the medicine of the present invention is in the form of granules or tablets, the dosage is converted into a decoction piece dosage and then taken orally.

[0016] The composition of the present invention can significantly improve the liver function of the mouse bile duct ligation-induced primary biliary cholangitis model, reduce the increase of AST and ALT in serum, and significantly reduce the gene expression of IL-33, MCP-1 and CXCL1. In the chemically induced mouse liver cholestasis model, the composition of the present invention can significantly reduce the serum AST, ALT and ALP levels; inhibit the gene expression of IL-33, MCP-1 and CXCL1 in liver tissue, indicating that the Chinese medicine composition can significantly reduce liver damage and inhibit inflammatory response, thereby playing a therapeutic role in primary biliary cholangitis. DETAILED DESCRIPTION

[0017] As mentioned above, the present invention aims to provide a use of a Chinese medicine composition in the preparation of a drug for primary biliary cholangitis. The following will be described in detail in conjunction with the content of a specific experiment.

[0018] The following experiments, if no specific conditions are specified, are conducted under conventional conditions or conditions recommended by the manufacturer. The raw materials or excipients used, as well as the reagents or instruments used, if the manufacturer is not specified, are all conventional products that can be purchased commercially. Unless otherwise specified, all percentages, ratios, proportions or parts are by weight. Unless otherwise specified, the medicinal materials of the present invention can be raw products or processed products thereof without significantly affecting the overall effect of the prescription.

[0019] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the present invention.

[0020] Example 1 Preparation of Chinese medicine composition

[0021] Take 10 parts of cooked aconite, 15 parts of dried ginger, 20 parts of roasted licorice, 10 parts of honeysuckle, 10 parts of Gleditsia sinensis, 20 parts of Ipomoea australis, 10 parts of Patchouli, and 5 parts of dried tangerine peel, add 8 times the total mass of water of the raw material medicine, heat to boiling, keep slightly boiling for 1 hour, and separate to obtain the first decoction and medicinal residue; for the second time, take the medicinal residue obtained after the first decoction, add 8 times the total mass of water of the raw material medicine, heat to boiling, keep slightly boiling for 1 hour, and separate to obtain the second decoction and medicinal residue; combine the first decoction and the second decoction to obtain a decoction, reduce the pressure and concentrate the decoction filtrate to obtain a clear paste, grind it into fine powder to obtain a traditional Chinese medicine composition, add maltodextrin, mix well, granulate, and obtain 1000g of granules to obtain traditional Chinese medicine composition granules.

[0022] Experimental Example 2 Therapeutic effect of Chinese medicine composition on primary biliary cholangitis model induced by bile duct ligation in mice

[0023] 1. Experimental Materials

[0024] 1.1 Animals

[0025] C57 / BL6J mice, SPF grade, 50 mice, body weight 18-22 g, male.

[0026] Rearing environment: room temperature 20-26℃, relative humidity controlled at 40-70%, 10 per group per cage.

[0027] 1.2 Drugs

[0028] The Chinese medicine composition prepared in Example 1 was purchased from Jiangsu Kangyuan Pharmaceutical Co., Ltd.; isoflurane was purchased from Shenzhen Ruiwode Life Science Co., Ltd.

[0029] 1.3 Instruments

[0030] Electronic balance (Sartorius Scientific Instruments, model BS224S); Electronic balance (G&G, model TC3K). Small animal gas anesthesia machine (Shenzhen Ruiwode Life Science Technology Co., Ltd., model R500).

[0031] 2. Dosage design

[0032] The daily dosage of the Chinese medicine composition is 100g of crude drug / day. According to the body surface area conversion method, the equivalent dose for mice is 100g / 60kg*12.3=20.50g of crude drug / kg. The low dose is set to 10.25g of crude drug / kg, the medium dose is set to 20.50g of crude drug / kg, and the high dose is set to 41.00g of crude drug / kg. Oral administration, 0.2mL / 10g, once a day.

[0033] 3. Experimental Methods

[0034] C57 / BL6J mice were adaptively fed for 7 days and were allowed to eat feed and water freely. They were randomly divided into sham operation control group, model control group, high-dose, medium-dose and low-dose groups of the Chinese medicine composition according to their body weight, with 10 mice in each group.

[0035] Before performing the bile duct ligation (BDL) surgery on mice, ensure that all instruments and equipment are sterilized. Keep the mice warm on a 37°C heating plate throughout the experiment. Induce anesthesia using a 4% isoflurane and air mixture until the mice are pain-free. Shave the abdominal hair, then fix the mice on a heating plate and maintain anesthesia with 1.5-3% isoflurane mixture inhalation. Clean the abdomen with iodine skin disinfectant.

[0036] The abdomen was opened with a 2-cm midline abdominal incision, and a retaining suture and retractor were inserted to expand the surgical area. The liver was lifted with a moistened cotton swab to expose the bile duct. The bile duct, portal vein, and hepatic artery were separated using micro-toothed forceps. Two surgical knots were placed consecutively on the bile duct to tie off the bile duct, taking care not to cut the bile duct. After flushing the abdominal cavity with normal saline, the organs were repositioned and the abdomen was sutured closed. The above modeling was completed in all model groups and drug-treated groups. The sham control group only opened the abdominal cavity, separated the bile duct, portal vein, hepatic vein, and sutured the abdomen.

[0037] After the animals recovered and were observed to be normal, the next day was regarded as the first day of administration. The corresponding doses of drugs were given for intervention, and the normal control group was dripped with an equal amount of saline. Each group was gavaged at 0.2mL / 10g each time, once a day, for 14 consecutive days. On the 14th day, 30 minutes after the administration of each group of mice, blood was taken for the detection of ALT and AST under anesthesia, and liver tissue was taken for the detection of IL-33, MCP-1 and CXCL1.

[0038] 4. Statistical methods

[0039] The quantitative indicators were first tested for variance homogeneity using the LEVENE test. When the variance was homogeneous (P>0.05), one-way analysis of variance (ANOVA) was used for statistical testing; when the variance was unequal (P≤0.05), the Kruskal-Wallis H rank sum test (KW method) was used for statistical analysis. When the one-way ANOVA showed that the difference was statistically significant (P≤0.05), Dunnett's t test (Dunnett method) was used to compare the differences between the groups; when the one-way ANOVA showed that the difference was not statistically significant (P>0.05), the statistical analysis was terminated. When the Kruskal-Wallis H rank sum test showed that the difference was statistically significant (P≤0.05), the Mann-Whitney U test (MW method) was used to compare the differences between the groups; when the Kruskal-Wallis H rank sum test showed that the difference was not statistically significant (P>0.05), the statistical analysis was terminated.

[0040] 5. Experimental Results

[0041] 5.1 Effects of serum AST and ALT in the mouse model of primary biliary cholangitis induced by bile duct ligation

[0042] After bile duct ligation in mice, the serum AST and ALT levels of mice were significantly increased compared with the normal control group, with statistical differences (P<0.001); 14 days after the start of treatment with the Chinese medicine composition, the high and medium dose AST and ALT levels were significantly reduced compared with the model control group, with statistical differences (P<0.001, (P<0.01). See Table 1.

[0043] Table 1: Effects on serum AST and ALT in the mouse model of primary biliary cholangitis induced by bile duct ligation

[0044]

[0045] Note: Compared with the normal control group, ### P<0.001; compared with the model control group, **P<0.01, ***P<0.001; data are expressed as Mean±SD.

[0046] 5.2 Effects on the expression levels of IL-33, MCP-1 and CXCL1 genes in liver tissue of the mouse model of primary biliary cholangitis induced by bile duct ligation

[0047] Compared with the normal control group, the expression levels of inflammatory factor IL-33, chemokine MCP-1 and CXCL1 in the liver tissue cells of the model group mice were significantly increased, with statistical differences (P<0.001); 14 days after the start of treatment with the Chinese medicine composition, compared with the model control group, the expression levels of high and medium doses of IL-33, MCP-1 and CXCL1 were significantly reduced, with statistical differences (P<0.001, P<0.01). See Table 2.

[0048] Table 2: Effects on the expression levels of IL-33, MCP-1 and CXCL1 genes in liver tissue of the mouse model of primary biliary cholangitis induced by bile duct ligation

[0049]

[0050] Note: Compared with the normal control group, ### P<0.001; compared with the model control group, **P<0.01, ***P<0.001; data are expressed as Mean±SD.

[0051] 6. Experimental conclusion

[0052] The Chinese medicine composition of 20.50 and 41.00 g crude drug / kg can significantly inhibit the serum AST and ALT levels in the primary biliary cholangitis model induced by bile duct ligation in mice, and reduce the gene expression levels of inflammatory factor IL-33 and chemokines MCP-1 and CXCL-1 in liver tissue, indicating that the Chinese medicine composition can effectively protect the liver and has a certain therapeutic effect on the primary biliary cholangitis model induced by bile duct ligation in mice.

[0053] Experimental Example 3 Protective effect of Chinese medicine composition on α-naphthalene isothiocyanate-induced liver cholestasis model in mice

[0054] 1. Experimental Materials

[0055] 1.1 Animals

[0056] C57 / BL6J mice, SPF grade, 50 mice, body weight 18-22 g, male.

[0057] Rearing environment: room temperature 20-26℃, relative humidity controlled at 40-70%, 10 per group per cage.

[0058] 1.2 Drugs

[0059] The Chinese medicine composition was prepared according to the method of Example 1 and provided by Jiangsu Kangyuan Pharmaceutical Co., Ltd.; α-naphthalene isothiocyanate, Sigma-Aldrich.

[0060] 1.3 Instruments

[0061] Electronic balance (Sartorius Scientific Instruments, model BS224S); electronic balance (G&G, model TC3K); low-temperature centrifuge (Eppendorf, Germany, model 5804R); microplate reader (MeiGu Molecular, USA, model Flexstation 3).

[0062] 2. Dosage design

[0063] The daily dosage of the Chinese medicine composition is 100g of crude drug / day. According to the body surface area conversion method, the equivalent dose for mice is 100g / 60kg*12.3=20.50g of crude drug / kg. The low dose is set to 10.25g of crude drug / kg, the medium dose is set to 20.50g of crude drug / kg, and the high dose is set to 41.00g of crude drug / kg. Oral administration, 0.2mL / 10g, once a day.

[0064] 3. Experimental Methods

[0065] C57 / BL6J mice were adaptively fed for 7 days with free access to feed and water. They were randomly divided into blank control group, model control group, high-dose, medium-dose and low-dose groups of Chinese medicine composition according to body weight, with 10 mice in each group. The high-dose, medium-dose and low-dose groups of Chinese medicine composition were given the test drug intervention for 6 consecutive days. The blank control group and the model control group were given an equal volume of corn oil. On the 4th day of administration, the model control group and the high-dose, medium-dose and low-dose groups of Chinese medicine composition were given α-naphthalene isothiocyanate 80 mg / kg by oral gavage. On the 6th day of the experiment, the animals were killed 30 minutes after the last administration. Blood was taken for the detection of ALT, AST and ALP, and liver tissue was taken for the detection of IL-33, MCP-1 and CXCL1.

[0066] 4. Statistical methods

[0067] The quantitative indicators were first tested for variance homogeneity using the LEVENE test. When the variance was homogeneous (P>0.05), one-way analysis of variance (ANOVA) was used for statistical testing; when the variance was unequal (P≤0.05), the Kruskal-Wallis H rank sum test (KW method) was used for statistical analysis. When the one-way ANOVA showed that the difference was statistically significant (P≤0.05), Dunnett's t test (Dunnett method) was used to compare the differences between the groups; when the one-way ANOVA showed that the difference was not statistically significant (P>0.05), the statistical analysis was terminated. When the Kruskal-Wallis H rank sum test showed that the difference was statistically significant (P≤0.05), the Mann-Whitney U test (MW method) was used to compare the differences between the groups; when the Kruskal-Wallis H rank sum test showed that the difference was not statistically significant (P>0.05), the statistical analysis was terminated.

[0068] 5. Experimental Results

[0069] 5.1 Effects of α-naphthalene isothiocyanate on serum AST, ALT and ALP in mice with hepatic cholestasis

[0070] Compared with the normal control group, the serum AST, ALT and ALP levels of the mice in the model group were significantly increased, with statistical differences (P<0.001); compared with the model control group, the high, medium and low doses of AST, ALT and ALP levels were significantly reduced, with statistical differences (P<0.001, P<0.01). See Table 3.

[0071] Table 3: Effects on serum AST, ALT and ALP in the mouse liver cholestasis model induced by α-naphthalene isothiocyanate

[0072]

[0073] Note: Compared with the normal control group, ### P<0.001; compared with the model control group, **P<0.01, ***P<0.001; data are expressed as Mean±SD.

[0074] Effects of α-naphthalene isothiocyanate on the gene expression levels of IL-33, MCP-1 and CXCL1 in liver tissue of mice with hepatic cholestasis

[0075] Compared with the normal control group, the gene expression of inflammatory factor IL-33, chemokine MCP-1 and CXCL1 in the liver tissue of mice was statistically different (P<0.001); on the 6th day after the start of treatment with the Chinese medicine composition, compared with the model control group, the gene expression of inflammatory factor IL-33, chemokine MCP-1 and CXCL1 in the liver tissue of high-, medium- and low-dose mice was significantly reduced, with statistical differences (P<0.001). See Table 4.

[0076] Table 4: Effects on the gene expression levels of IL-33, MCP-1 and CXCL1 in liver tissue of α-naphthalene isothiocyanate-induced mouse liver cholestasis model

[0077]

[0078]

[0079] Note: Compared with the normal control group, ### P<0.001; compared with the model control group, **P<0.01, ***P<0.001; data are expressed as Mean±SD.

[0080] 6. Experimental conclusion

[0081] The Chinese medicine composition can significantly inhibit the increase of AST, ALT and ALP in the serum of the mouse liver cholestasis model induced by α-naphthalene isothiocyanate, and reduce the gene expression of IL-33, MCP-1 and CXCL1 in liver tissue, indicating that the Chinese medicine composition can effectively protect the liver and has a certain therapeutic effect on the mouse liver cholestasis model induced by α-naphthalene isothiocyanate.

[0082] The above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the embodiments here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. Use of a Chinese medicine composition in the preparation of a drug for primary biliary cholangitis, characterized in that: The traditional Chinese medicine composition is prepared from the following components: 10-30 parts of aconite root, 5-15 parts of dried ginger, 20-30 parts of liquorice, 10-20 parts of honeysuckle, 10-30 parts of Gleditsia sinensis, 20-45 parts of Ipomoea australis, 10-15 parts of patchouli, and 5-15 parts of tangerine peel.

2. The use according to claim 1, characterized in that: The invention is prepared from the following raw material components by weight: 10-20 parts of aconite root, 10-15 parts of dried ginger, 20-25 parts of liquorice, 10-15 parts of honeysuckle, 10-20 parts of gleditsia thorn, 20-30 parts of cinnamon bark, 10-12 parts of patchouli and 5-10 parts of tangerine peel.

3. The use according to claim 1, characterized in that: The traditional Chinese medicine composition is prepared from the following components by weight: 10-12 parts of aconite root, 12-15 parts of dried ginger, 20-22 parts of liquorice, 10-12 parts of honeysuckle, 10-12 parts of Gleditsia sinensis, 20-22 parts of Ipomoea australis, 10-12 parts of patchouli, and 5-6 parts of tangerine peel.

4. The use according to claim 1, characterized in that: The invention is prepared from the following raw material components by mass: 10 parts of aconite root, 15 parts of dried ginger, 20 parts of liquorice, 10 parts of honeysuckle, 10 parts of gleditsia thorn, 20 parts of cinnamon bark, 10 parts of patchouli, and 5 parts of tangerine peel.

5. The use according to any one of claims 1 to 4, characterized in that: The drug also includes pharmaceutically acceptable excipients or additives medically acceptable excipients.

6. The use according to claim 5, characterized in that The pharmaceutically acceptable excipients or additives include fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, preservatives, and bases.

7. The application according to claim 6, characterized in that: The dosage form for the drug includes an oral dosage form, an injection dosage form or an external dosage form.

8. The use according to claim 4, characterized in that: The pharmaceutical composition is in the form of granules, tablets, capsules, pills, oral solutions, oral suspensions or oral emulsions.

9. The use according to claims 1-4, characterized in that: The preparation method of the traditional Chinese medicine composition comprises the following steps: taking aconite root, dried ginger, liquorice, honeysuckle, thorn of Gleditsia sinensis, five-clawed dragon, patchouli, and tangerine peel, adding 8 to 12 times of water equivalent to the total mass of the raw materials, heating to boiling, maintaining a slight boiling for 1 to 2 hours, and obtaining a first decoction and medicinal residues after separation; a second time, taking the medicinal residues obtained after the first decoction, adding 6 to 10 times of water equivalent to the total mass of the raw materials, heating to boiling, maintaining a slight boiling for 1 to 2 hours, and obtaining a second decoction and medicinal residues after separation; combining the first decoction and the second decoction to prepare a decoction, and reducing the pressure on the decoction filtrate to concentrate to obtain a clear paste, and crushing it.

10. Use of a Chinese medicine composition in the preparation of a liver protection drug, characterized in that: The traditional Chinese medicine composition is prepared from the following components: 10-30 parts of aconite root, 5-15 parts of dried ginger, 20-30 parts of liquorice, 10-20 parts of honeysuckle, 10-30 parts of Gleditsia sinensis, 20-45 parts of Ipomoea australis, 10-15 parts of patchouli, and 5-15 parts of tangerine peel.

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