CN111297859A - Application of koumine in preparation of medicine for treating liver cell injury - Google Patents
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/475—Quinolines; Isoquinolines having an indole ring, e.g. yohimbine, reserpine, strychnine, vinblastine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
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- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
Abstract
The invention relates to the technical field of medicines, and provides application of koumine or pharmaceutically acceptable salts thereof in preparation of medicines for treating liver cell injury. Further, the pharmaceutical composition for treating liver cell injury is composed of an active ingredient and pharmaceutically acceptable auxiliary materials, wherein the active ingredient is koumine or pharmaceutically acceptable salts thereof, or the combination of the koumine or the pharmaceutically acceptable salts thereof and one or more other active ingredients. Through pharmacological experiments, the gelsemium elegans can improve the activity of normal and aflatoxin-exposed human liver L02 cells at a lower concentration; through transcriptomic analysis, the koumine can improve the expression of genes related to the growth of liver cells, so that the effect of promoting the growth of the liver cells is realized, for example, the koumine can obviously improve the transcription levels of genes related to the division, proliferation and growth functions of L02 cells, such as EGR3 genes, CCN2 genes and CCN1 genes. Therefore, a new theoretical basis is provided for the gelsemium or the salt thereof in the aspect of treating the hepatic cell injury.
Description
Technical Field
The invention belongs to the field of biological medicines, and particularly relates to application of koumine or pharmaceutically acceptable salts thereof in preparation of medicines for treating liver cell injury, and a medicine composition containing the koumine.
Background
The liver is not only the most important nutrient metabolism organ of the human body, but also the biotransformation organ of many non-nutrients such as drugs, poisons and toxic metabolites, and thus is susceptible to various substances ingested into the body, causing damage. It is known that various liver pathogenic factors, such as virus and bacterial infection, chemical poison and drug intake, immune dysfunction, ischemia reperfusion injury and the like all cause hepatocyte damage and liver function damage, so that the liver can not play a normal function, and further a series of symptoms are caused, even acute and chronic liver failure which is difficult to cure is developed. The action of the liver in human body is very important, once damaged, the functions of metabolism, detoxification, immunity and the like are influenced, the life quality of patients is seriously influenced, and even death is caused (Zhao Guang. research on new protective measures for liver damage, Wuhan, China university of science and technology, 2017). In response to liver injury, although many novel therapeutic drugs have been developed clinically at present, it is difficult to achieve effective treatment.
The virulent plant Gelsemium elegans Benth, also called Gelsemium elegans Benth, kudzu, big tea and the like, is widely distributed in southern provinces such as Fujian, Zhejiang, Guangxi, Guangdong, Yunnan and Guizhou and places such as southeast Asia in China, and has abundant resources (Liuhao, Shuchangxi, Gelsemium research progress, university of Fujian medical science, 2008,42(5): 469-. Although gelsemium has strong toxicity, it has many effects of dispelling pathogenic wind, counteracting toxic substance, subsiding swelling, relieving pain, etc., and can be used for treating scabies, eczema, scrofula, carbuncle swelling, furuncle, traumatic injury, rheumatalgia, neuralgia, etc., and has long history of application and broad prospect.
Pharmacological research proves that Gelsemium alkaloid has the effects of calming, easing pain, regulating immunity, inhibiting tumor growth and the like, and has better effects of clinically applying the Gelsemium alkaloid to the treatment of various pains such as cancer pain, sciatica, smooth muscle colic and the like and skin diseases such as eczema, furuncle, carbuncle, psoriasis and the like [ Jin GL, SuYP, Liu M, xuY, Yang J, Liao KJ, YuCX. medicinal plants Gelsemiium (Gelsemiaceae, Gentianales) -Ariewoofheir phytochemistry, pharmacoly, toxity dtradition use, JEthnopharma ]. However, the gelsemium elegans has strong toxicity and complex components, so that poisoning is easily caused during clinical application, and even death of patients can be caused.
In order to avoid toxic and side effects, the prior medicinal application mainly comprises pounding or grinding and applying the traditional Chinese medicines, and other external therapies (Jiangsu New medical college, Chinese medicine dictionary, Shanghai science and technology Press). Therefore, the alkaloid component extracted from the gelsemium elegans is separated and purified, the biological activity of the alkaloid monomer is detected, the toxic and side effects caused by clinically applying the gelsemium elegans crude extract containing various complex components to treat diseases are reduced, the application value of the gelsemium elegans crude extract is enhanced, and a space is provided for the research and development of new drugs.
Koumine (Koumine) is an indole alkaloid with the highest content and lower toxicity in gelsmium elegans. Chinese patent CN101323618B discloses a method for extracting and separating koumine from gelsemium elegans, but does not describe the new use thereof. At present, the prevention and treatment effects of koumine in diseases or symptoms related to hepatocyte injury are not reported in documents.
Disclosure of Invention
The invention aims to provide a new medical application of koumine or a salt thereof and a pharmaceutical composition containing the koumine.
The first aspect of the invention provides the application of koumine or its pharmaceutically acceptable salt in preparing a medicament for treating liver cell injury.
The koumine has the following structural formula, and the molecular formula is C20H22N2O, which is either obtainable from the alkaloid monomer of gelsemium elegans or synthetically obtainable:
the pharmaceutically acceptable salt of koumine comprises salt of koumine with inorganic acid or organic acid. Exemplary pharmaceutically acceptable salts include, but are not limited to: sulfate, hydrochloride, hydroiodide, hydrobromide, nitrate, phosphate, bisulfate, citrate, acid phosphate, oxalate, acetate, lactate, isonicotinate, acid citrate, salicylate, oleate, tartrate, pantothenate, tannate, bitartrate, succinate, ascorbate, maleate, fumarate, gentisate, glucuronate, gluconate, benzoate, formate, methanesulfonate, glutamate, benzenesulfonate, ethanesulfonate, pamoate, and p-toluenesulfonate. Preferably, the pharmaceutically acceptable salts are koumine sulfate and hydrochloride.
Further, the liver cell damage includes liver cell damage caused by liver cirrhosis, liver malignant tumor, hepatotoxic drug or exposure to antituberculosis agent or chemotherapeutic agent, and the specific symptoms include viral hepatitis, autoimmune hepatitis, drug hepatitis, toxic hepatitis, alcoholic hepatitis, metabolic hepatitis and liver cirrhosis and liver cancer developed by the same. Especially the liver cell damage caused by aflatoxin.
In order to verify the protection effect of gelsemium elegans on liver cell injury, the invention verifies the protection effect of gelsemium elegans on human liver cells L02 through cytology in vitro experiments: an aflatoxin AFB1 exposed cell model is prepared by using normal human liver cell L02 cells, and then koumine with the concentrations of 5 mu M and 20 mu M is co-cultured with the koumine. Pharmacological experiments show that koumine can improve the activity of L02 cells of normal and AFB1 exposed human livers at the concentrations of 5 mu M and 20 mu M (shown in figure 1); through transcriptomic analysis, the koumine can improve the expression of genes related to the growth of liver cells, so that the effect of promoting the growth of the liver cells is realized, for example, the koumine can obviously improve the transcription levels of genes related to the division, proliferation and growth functions of L02 cells, such as EGR3 genes, CCN2 genes and CCN1 genes.
In addition, toxicity tests show that the LD50 of the gelsemium is 91.25mg/kg by intraperitoneal injection of mice, which is far lower than the toxicity of the total alkaloids of gelsemium reported in the literature.
Therefore, the medicine for treating the hepatocyte damage is a medicine for improving the hepatocyte activity or a medicine for promoting the expression of hepatocyte growth related genes EGR3, CCN1 or CCN 2.
In a second aspect of the present invention, a pharmaceutical composition for treating liver cell injury is provided, which comprises an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient is koumine or its pharmaceutically acceptable salt as the only active ingredient, or is a combination of koumine or its pharmaceutically acceptable salt and one or more other active ingredients.
The one or more additional active ingredients are one or more biotherapeutic agents, chemotherapeutic agents, or combinations thereof known in the art to have therapeutic efficacy for the above-mentioned diseases. By using in combination with gelsemine, effects including, but not limited to, the following can be achieved: reducing the dosage of other active ingredients to avoid side effects; the overall curative effect brought by each active ingredient is improved; maintaining the therapeutic efficacy.
The one or more further active ingredients may be selected from:
anti-inflammatory liver-protecting drugs: compound glycyrrhizin, diammonium glycyrrhizinate, magnesium isoglycyrrhizinate or bicyclol;
enzyme-reducing liver-protecting medicine: biphenyldicarboxylate, oleanolic acid, Yinzhihuang;
detoxification type liver-protecting drugs: glutathione, tiopronin, glucuronolactone;
liver-protecting drugs for promoting hepatocyte repair: polyene phosphatidyl choline, hepatocyte growth promoting factor, silymarin;
cholagogic hepatoprotective agents: ademetionine, ursodeoxycholic acid, anethol trithione.
In combination therapy, the gelsemium or a pharmaceutically acceptable salt thereof and the other active ingredients may be administered to the individual simultaneously or sequentially. The combined administration includes simultaneous or separate administration using compositions containing gelsemium or a pharmaceutically acceptable salt thereof and other active ingredients, respectively, or administration of a single composition containing the above active ingredients.
As used herein, "individual" refers to a mammal, including, but not limited to, a primate (e.g., a human, gorilla, chimpanzee, etc.), a rodent (e.g., mouse, rat, hamster, gerbil), a lagomorpha, a porcine (e.g., jorkshire, xianghe), a canine, a feline, an equine, etc., preferably a human.
By "treatment" is meant the complete or partial cure or elimination of a disease, including but not limited to one or a combination of more of the following: alleviating or eliminating the cause of the disease, ameliorating or eliminating pathological changes, alleviating or eliminating symptoms thereof, slowing or arresting progression, reducing relapse, reducing morbidity thereof, and improving prognosis thereof.
The pharmaceutically acceptable adjuvants include diluent, excipient, binder, disintegrating agent, lubricant, glidant, antiseptic, pH regulator, suspending agent, dispersant, emulsifier, ointment matrix, penetration enhancer, emollient, propellant, surfactant, sweetener, flavoring agent or drug release regulator, etc. One skilled in the art can select an appropriate carrier according to the desired formulation form, drug release characteristics, administration mode, etc. of the drug.
The medicament may be formulated into dosage forms in solid, semi-solid, or liquid form suitable for administration by any recognized mode of administration available in the art, such as administration, including, but not limited to: suitable for oral administration, such as tablets, powders, capsules, granules, syrups, troches, solutions or suspensions and the like; sterile solutions or suspensions suitable for parenteral administration, e.g. intramuscular, subcutaneous or intravenous injection; plasters, ointments, creams, patches, sprays or gels suitable for topical administration, e.g. through the skin or mucosa; suitable for vaginal or rectal administration, such as suppository, gel, effervescent tablet or emulsion.
As used herein, a "therapeutically effective amount" refers to the amount of active ingredient that will achieve the desired therapeutic effect for the use or method described.
In the uses or methods described herein, the dosage of koumine or a pharmaceutically acceptable salt thereof and/or other active ingredients will depend on a variety of factors, including the severity of the condition, the individual characteristics being treated, the mode of administration and the judgment of the practitioner. Generally, an effective dose is from about 0.0001 to about 50mg per kg body weight per day (single or divided administration). For a 60kg human, from about 0.006 mg/day to about 3000 mg/day. In some cases, dosage levels not higher than the lower limit of the aforesaid range may be sufficient, while in other cases still larger doses may be administered without causing any harmful effects, and larger doses may be divided into multiple small doses for administration over 24 hours, if necessary.
The amount of gelsemium or a pharmaceutically acceptable salt and/or other active ingredient thereof in the medicament or in the method of treatment may be from about 0.001mg to about 500mg, for example from about 0.02 mg to about 300mg, or from about 1mg to about 20mg, preferably from about 0.1 mg to about 20mg, for example 0.2mg, 1mg, 3mg, 5mg, 10mg, 15mg or 20mg etc.
Action and Effect of the invention
Through pharmacological experiments, the gelsemium elegans can improve the activity of normal and aflatoxin-exposed human liver L02 cells at a lower concentration; through transcriptomic analysis, the koumine can improve the expression of genes related to the growth of liver cells, so that the effect of promoting the growth of the liver cells is realized, for example, the koumine can obviously improve the transcription levels of genes related to the division, proliferation and growth functions of L02 cells, such as EGR3 genes, CCN2 genes and CCN1 genes. Therefore, the invention provides a new theoretical basis for the gelsemium or the salt thereof in the aspect of treating the hepatic cell injury.
Drawings
FIG. 1 shows the effect of Gelsemiq on the viability of L02 cells in normal and AFB1 exposed human liver.
Detailed Description
The present invention will be described in detail below with reference to examples and the accompanying drawings. The following examples should not be construed as limiting the scope of the invention.
Example 1: protective effect of koumine on exposure of human liver cell L02 cell to aflatoxin
An aflatoxin (AFB1) exposed cell model is prepared by using normal human liver cell L02 cells:
(1) l02 cells were cultured in RPMI-1640 medium (90%) + FBS (10%) + double antibody (1%), and passaged once every 2-3 days in a 37 ℃ culture environment containing 5% CO2An incubator, when the density reaches more than 80%, carrying out passage with the passage ratio of 1:2, and selecting cells between 10 generations and 40 generations for experiment;
(2) 5mg of AFB1 was dissolved in 250. mu.L of dimethyl sulfoxide sufficiently to give a stock solution of 20mg/ml, which was then diluted to a concentration of 10. mu.M in RPMI-1640 medium;
(3) dissolving 2mg of gelsemium elegans in 100 μ L of dimethyl sulfoxide to obtain 20mg/mL stock solution, and diluting with RPMI-1640 culture medium to obtain 20 μ M and 5 μ M solutions;
(4) l02 cells were cultured at 1X 105The culture medium was inoculated in a density of 100. mu.L per well in a 96-well plate, and after culturing for 24 hours to allow adherence, the culture medium was replaced.
The cells were divided into a blank control group, a gelsemium-treated group and a negative control group, wherein 100. mu.L of fresh medium was added to each well of the blank control group, and 100. mu.L of fresh medium containing 10. mu.MAFB 1 was added to each well of the gelsemium-treated group and the negative control group. After 3 hours of culture, all the culture media are discarded, 100 mu L of fresh culture media are added into each hole of the blank control group and the negative control group, and fresh culture media containing 20 mu M and 5 mu M of gelseminal koumine are respectively added into the gelsemine treatment group for culture.
After 33 hours, all groups are discarded, 100 mu L of fresh culture medium and 10 mu L of cck-8 reagent are added into each hole, the mixture is fully mixed after each hole is added, the mixture is continuously incubated in an incubator for 1 hour, then the hole plate is taken out, and the absorbance at 450nm is detected by using an enzyme-labeling instrument. According to the formula: the cell survival rate is ═ 100% of [ (absorbance of laboratory test wells-absorbance of blank control wells)/(absorbance of control wells-absorbance of blank control wells) ], and the cell survival rate was calculated.
The results are shown in FIG. 1, and koumine increased the viability of normal and AFB 1-exposed human liver L02 cells at both 20. mu.M and 5. mu.M concentrations.
Example 2 Gelsemiin increases the expression level of growth-related genes of human liver L02
An aflatoxin AFB1 exposed cell model was made using the method in example 1. After 33 hours, the culture medium was discarded from the blank control group, the gelsemium treated group and the negative control group, washed twice with 100. mu.L/well of PBS, the residual medium was removed, 50. mu.L of LTRIzol was added to each well, and the cells were collected in an EP tube after they were lysed thoroughly by repeated pipetting.
Firstly, RNA extraction:
A. taking a proper amount of cell sample, adding 600 mu L of lysine/Binding Buffer, and homogenizing; adding 30 μ L mirnaeogenin additive, mixing well and ice-cooling for 10 min; B. adding phenol or chloroform with the same volume, centrifuging at the maximum rotation speed at room temperature for 5min, and collecting the supernatant; C. adding 1.25 times volume of 100% ethanol, adding the mixture into a centrifugal column (700 μ L can be added at the maximum), standing at room temperature at 13000rpm for 30s, and discarding the supernatant; D. adding 350 μm of the ImiRNAWash Solution 1 to the spin column at 13000rpm for 30s, discarding the supernatant, and replacing the spin column into the collecting tube; E. mix 10 μ LDNase I and 70 μ LBuffer RDD QIAGEN (#79254) to a total volume of 80 μ L, apply to the membrane in the spin column, and stand at room temperature for 15 min; F. add 350. mu.L of mirnavashsolution 1 to the spin column, 13000rpm, 30 s. Discarding the supernatant, and replacing the spin column into the collecting pipe; g.500. mu.L of the liquid Solution 2/3 was passed through the column twice at 13000rpm for 30 s. Discarding the supernatant, and replacing the spin column into the collecting pipe; H. centrifuging the empty column for 1 min, placing the column into a new collection tube, adding 100 μ L of 95 deg.C preheated Elutionsolution at the center of the column, standing for 2min, centrifuging at room temperature and highest rotation speed for 20-30 s, and collecting the liquid in the collection tube as the extracted Total RNA, which can be stored at-70 deg.C.
II, constructing a library by chain specificity transcriptome:
(1) purification and fragmentation of mRNA: mixing 50 μ L total RNA (4 μ g) and 50 μ L RNAPropertyBeads, cooling to 4 deg.C on PCR instrument at 65 deg.C for 5min, standing at room temperature for 5min, standing on magnetic frame for 5min until supernatant is clarified, and discarding supernatant. Taking down the magnetic frame, adding 200 μ LBeads Washing Buffer, mixing, standing on the magnetic frame for 5min until the supernatant is clarified, removing the supernatant, adding 50 μ L of precipitation Buffer, mixing, placing on a PCR instrument at 80 deg.C for 2min, and cooling to 25 deg.C; adding 50 μ LBeads Binding Buffer, blowing for 6 times, mixing, standing at room temperature for 5min, standing on magnetic frame for 5min until the supernatant is clear, and removing the supernatant; taking down the magnetic frame, adding 200 μ LBeads Washing Buffer, blowing and beating for 6 times, mixing, standing on the magnetic frame for 5min until the supernatant is clear, and discarding the supernatant; taking down the magnetic frame, adding 19.5 μ L of Elute, Primer, FragmentMix, beating for 6 times, mixing, cooling to 4 deg.C for 8min at 94 deg.C on PCR instrument, taking out, and centrifuging for a short time.
(2) Synthesis of cDNA 1 Strand: adding 8 μ LFirst Strand Synthesis surface enzyme D Mix and SuperScript II Reverse Transcriptase, beating for 6 times, mixing, cooling to 4 deg.C in PCR instrument at 25 deg.C, 10min, 42 deg.C, 15min, 70 deg.C, 15min, and taking out.
(3) Synthesis of cDNA2 Strand: after 5. mu.L of End Repair Control (2. mu.L of End Repair Control + 98. mu.L of Resuspension Buffer) was added, 20. mu.L of Second Strand Marking Master Mix was added, and the mixture was whipped 6 times, mixed well, placed on a PCR instrument at 16 ℃ for 60min, and taken out. Adding 90 μ L of AMPure XPbeads, blowing for 10 times, mixing, standing at room temperature for 15min, standing on a magnetic frame for 5min until the supernatant is transparent, sucking 135 μ L of supernatant with 200 μ L of suction head, and discarding; the plate was placed on a magnetic rack all the time, 200. mu.L of freshly prepared 80% ethanol was added, the temperature was 30s, the supernatant was discarded, the operation was repeated once and then placed at room temperature for 15min, air dried, the plate was removed from the magnetic rack, 17.5. mu.L of Resuspension Buffer was added, the mixture was blown up 10 times, after thorough mixing, the plate was placed on the magnetic rack for 5min until the supernatant was clear, and 15. mu.L of supernatant was transferred to a new PCR plate.
(4) 3' end with A: adding 2.5 μ L diluted A-labeling Control (1 μ LA-labeling Control +99 μ LResuspension Buffer), adding 12.5 μ LA-labeling Mix, beating 10 times, mixing, placing on PCR instrument at 37 deg.C for 30min, 70 deg.C for 5min, and cooling to 4 deg.C.
(5) Ligation of linker sequences: adding 2.5 μ L diluted Ligation Control (1 μ L Ligation Control +99 μ L LResusperation Buffer), adding 2.5 μ L Ligation Mix, 2.5 μ L LRNAAdapter index, beating 10 times, mixing, placing on PCR instrument at 30 deg.C for 10min, and taking off from the PCR instrument; adding 5 μ L of Stop ligation buffer, beating by blowing for 10 times, mixing well, adding 42 μ L of AMPure XPbeads, beating by blowing for 10 times, mixing well, standing at room temperature for 15min, placing on a magnetic frame until the supernatant is transparent, sucking 79.5 μ L of supernatant, and discarding. The plate was placed on a magnetic stand all the time, 200. mu.L of freshly prepared 80% ethanol was added, the temperature was 30s at room temperature, the supernatant was discarded, and the procedure was repeated once. Placing for 15min at room temperature, air drying, taking down the flat plate from the magnetic frame, adding 52.5 mu LResus suspension Buffer, blowing and beating for 10 times, fully mixing uniformly, placing for 2min at room temperature, placing on the magnetic frame for 5min until the supernatant is transparent, transferring 50 mu L of supernatant to a new PCR plate, adding 50 mu L of ampureXPbeads fully mixed uniformly, blowing and beating for 10 times, mixing uniformly, placing for 15min at room temperature, placing on the magnetic frame for 15min until the supernatant is transparent. Aspirate 95. mu.L of supernatant and discard. The plate was placed on a magnetic stand all the time, 200. mu.L of freshly prepared 80% ethanol was added, the temperature was 30s at room temperature, the supernatant was discarded, and the procedure was repeated once. Standing at room temperature for 15min, air drying, taking the plate off the magnetic frame, adding 22.5 μ LResuspesion Buffer, beating 10 times, and mixing well. After placing on a magnetic rack for 5min at room temperature for 2min until the supernatant is clear, 20. mu.L of the supernatant is transferred to a new PCR plate.
(6) Enrichment of DNA fragments: add 5. mu.L of PCR Primer Cocktail, add 25. mu.L of LPCR Master Mix, Mix well, PCR after brief centrifugation: 30s at 98 ℃, 15 cycles: 10s at 98 ℃; 30s at 60 ℃; 30s at 72 ℃; 5min at 72 ℃; hold at 10 ℃.
(7) And (3) purification: adding 50 μ L of AMPure XPbeads, beating for 10 times, mixing, standing at room temperature for 15min, standing on magnetic frame for 15min to obtain supernatant, sucking 95 μ L of supernatant, and discarding. The plate was placed on a magnetic stand all the time, 200. mu.L of freshly prepared 80% ethanol was added, the temperature was 30s at room temperature, the supernatant was discarded, and the procedure was repeated once. Standing at room temperature for 15min, air-drying, removing the plate from the magnetic rack, adding 32.5 μ L of LResuspesion Buffer, pipetting 10 times, mixing well, standing at room temperature for 2min, standing on the magnetic rack for 5min until the supernatant is clear, and transferring 30 μ L of the supernatant to a new PCR plate.
(8) Performing quality inspection on the library: the sample 1 was added to the Agilent 2100 bioanalyzer and the length and mass of the library were confirmed from the results.
The results are summarized in table 1:
TABLE 1 Effect of Gelsemiin on transcription level of L02 cell-associated genes
As can be seen from table 1, gelsemium significantly increases the transcription levels of genes having the functions of promoting cell division, proliferation and growth, such as EGR3, CCN2 and CCN1 genes, in human liver L02 cells, thereby achieving the effect of promoting liver cell growth.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are given by way of illustration of the principles of the present invention, and that various changes and modifications may be made without departing from the spirit and scope of the invention as defined by the appended claims. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (9)
1. Application of koumine or its pharmaceutically acceptable salt in preparing medicine for treating liver cell injury is provided.
2. The use of koumine or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of liver cell damage according to claim 1 wherein:
wherein the structural formula of the koumine is as follows:
the pharmaceutically acceptable salt is selected from the group consisting of sulfate, hydrochloride, hydroiodide, hydrobromide, nitrate, phosphate, bisulfate, citrate, acid phosphate, oxalate, acetate, lactate, isonicotinate, acid citrate, salicylate, oleate, tartrate, pantothenate, tannate, bitartrate, succinate, ascorbate, maleate, fumarate, gentisate, glucuronate, gluconate, benzoate, formate, methanesulfonate, glutamate, benzenesulfonate, ethanesulfonate, pamoate, or p-toluenesulfonate.
3. The use of koumine or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of liver cell damage according to claim 1 wherein:
wherein said hepatocyte injury comprises hepatocyte injury due to cirrhosis, liver malignancy, hepatotoxic drugs or exposure to anti-tubercular agents or chemotherapeutic agents.
4. The use of koumine or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of liver cell damage according to claim 1 wherein:
wherein the hepatocyte damage is hepatocyte damage caused by aflatoxin.
5. The use of koumine or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of liver cell damage according to claim 1 wherein:
wherein, the medicine for treating the hepatocyte damage is a medicine for improving the hepatocyte activity or a medicine for promoting the expression of hepatocyte growth related genes EGR3, CCN1 or CCN 2.
6. Use of koumine or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of liver cell damage according to any one of claims 1 to 4 wherein:
wherein the medicament for treating the hepatocyte injury is a medicament which takes the koumine or the pharmaceutically acceptable salt thereof as the only active ingredient or a medicament composition containing the koumine or the pharmaceutically acceptable salt thereof.
7. A pharmaceutical composition for treating hepatocyte injury is characterized by comprising an active ingredient and pharmaceutically acceptable auxiliary materials, wherein the active ingredient is koumine or pharmaceutically acceptable salts thereof, or the combination of the koumine or the pharmaceutically acceptable salts thereof and one or more other active ingredients.
8. The pharmaceutical composition for treating liver cell damage according to claim 7, wherein:
wherein the other active ingredients comprise one or more of compound glycyrrhizin, diammonium glycyrrhizinate, magnesium isoglycyrrhizinate or bicyclol, bifendate, oleanolic acid, Yinzhihuang, glutathione, tiopronin, glucuronolactone, polyene phosphatidyl choline, hepatocyte growth-promoting factor, silymarin, ademetionine, ursodeoxycholic acid, anethol trithione and S-ademetionine.
9. The pharmaceutical composition for treating liver cell damage according to claim 7, wherein:
wherein the pharmaceutically acceptable adjuvants are diluents, excipients, binders, disintegrants, lubricants, glidants, preservatives, pH regulators, suspending agents, dispersants, emulsifiers, ointment bases, penetration enhancers, emollients, propellants, surfactants, sweeteners, flavoring agents or drug release regulators.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112137996A (en) * | 2020-10-23 | 2020-12-29 | 湖南农业大学 | Medicine for preventing gelsemium poisoning |
| CN113755440A (en) * | 2021-09-10 | 2021-12-07 | 广东海洋大学 | AFB (active carbon boron)1Cell model establishing method for inducing chicken liver injury |
Citations (1)
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| CN109893528A (en) * | 2017-12-07 | 2019-06-18 | 福建医科大学 | Koumine treats the purposes of disorders of lipid metabolism and its related disease or illness |
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| CN109893528A (en) * | 2017-12-07 | 2019-06-18 | 福建医科大学 | Koumine treats the purposes of disorders of lipid metabolism and its related disease or illness |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112137996A (en) * | 2020-10-23 | 2020-12-29 | 湖南农业大学 | Medicine for preventing gelsemium poisoning |
| CN112137996B (en) * | 2020-10-23 | 2023-08-25 | 湖南农业大学 | Medicament for preventing gelsemium intoxication |
| CN113755440A (en) * | 2021-09-10 | 2021-12-07 | 广东海洋大学 | AFB (active carbon boron)1Cell model establishing method for inducing chicken liver injury |
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