CN113181150A - Application of SR1078 in preparation of drug for treating alcoholic liver injury and pharmaceutical composition - Google Patents
Application of SR1078 in preparation of drug for treating alcoholic liver injury and pharmaceutical composition Download PDFInfo
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- CN113181150A CN113181150A CN202110471216.6A CN202110471216A CN113181150A CN 113181150 A CN113181150 A CN 113181150A CN 202110471216 A CN202110471216 A CN 202110471216A CN 113181150 A CN113181150 A CN 113181150A
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- 206010067125 Liver injury Diseases 0.000 title claims abstract 8
- 231100000753 hepatic injury Toxicity 0.000 title claims abstract 8
- 230000001476 alcoholic effect Effects 0.000 title claims abstract 7
- 239000003814 drug Substances 0.000 title claims abstract 6
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract 6
- 229940079593 drug Drugs 0.000 title 1
- 239000004480 active ingredient Substances 0.000 claims abstract 2
- 239000002671 adjuvant Substances 0.000 claims 1
- 239000008187 granular material Substances 0.000 claims 1
- 238000002347 injection Methods 0.000 claims 1
- 239000007924 injection Substances 0.000 claims 1
- 239000003826 tablet Substances 0.000 claims 1
- 208000022309 Alcoholic Liver disease Diseases 0.000 abstract 5
- 241000699666 Mus <mouse, genus> Species 0.000 abstract 3
- 210000002472 endoplasmic reticulum Anatomy 0.000 abstract 2
- 210000004185 liver Anatomy 0.000 abstract 2
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 abstract 1
- 108010082126 Alanine transaminase Proteins 0.000 abstract 1
- 108010003415 Aspartate Aminotransferases Proteins 0.000 abstract 1
- 102000004625 Aspartate Aminotransferases Human genes 0.000 abstract 1
- 102100023580 Cyclic AMP-dependent transcription factor ATF-4 Human genes 0.000 abstract 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract 1
- 101000905743 Homo sapiens Cyclic AMP-dependent transcription factor ATF-4 Proteins 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 abstract 1
- 230000037396 body weight Effects 0.000 abstract 1
- 230000006372 lipid accumulation Effects 0.000 abstract 1
- 210000002966 serum Anatomy 0.000 abstract 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/16—Amides, e.g. hydroxamic acids
- A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
- A61K31/167—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide having the nitrogen of a carboxamide group directly attached to the aromatic ring, e.g. lidocaine, paracetamol
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- 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
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- Animal Behavior & Ethology (AREA)
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- Gastroenterology & Hepatology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
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Abstract
The invention relates to the technical field of new application of a medicament, in particular to application of SR1078 in preparing a medicament for treating alcoholic liver injury and a pharmaceutical composition. The embodiment of the invention proves that: SR1078 improves the liver/body weight ratio of alcoholic liver disease mice; SR1078 can improve the serum enzymology index of the mouse, make alcoholic liver disease mouse glutamic-pyruvic transaminase (ALT), glutamic-oxaloacetic transaminase (AST) content reduce apparently; SR1078 can improve endoplasmic reticulum stress condition of liver of alcoholic liver disease mouse, ROR alpha is raised, and key protein ATF4 and CHOP of endoplasmic reticulum stress are reduced; SR1078 can improve liver injury and lipid accumulation caused by alcohol. Provides a new strategy for treating alcoholic liver diseases, has important clinical treatment significance, shows that SR1078 can be used as an active ingredient of a medicament for treating alcoholic liver injury, and provides scientific basis for developing the medicament for treating alcoholic liver diseases in the future.
Description
Technical Field
The invention relates to the technical field of new application of a medicament, in particular to application of SR1078 in preparing a medicament for treating alcoholic liver injury and a pharmaceutical composition.
Background
China is the country with the most deaths caused by drinking, and the burden of diseases caused by drinking is also aggravated. In China, the number of alcoholic liver disease patients accounts for 4.5 percent of the total number of patients, and reaches 6200 ten thousand. Alcoholic Liver Disease (ALD) has three major stages: alcoholic fatty liver disease, alcoholic hepatitis and cirrhosis. Alcoholic fatty liver disease, characterized by the accumulation of fatty acids in the liver, is usually asymptomatic and reversible after alcohol withdrawal. Although most severe drinkers exhibit some level of fatty liver disease, in some cases, heavy drinking need only occur daily for a period of less than a week, with only one of five severe drinkers developing alcoholic hepatitis, and one of four developing cirrhosis. Alcoholic hepatitis is characterized by inflammation of liver cells and, in general, is reversible by alcohol withdrawal. Cirrhosis is generally irreversible and is characterized by inflammation, fibrosis (cell sclerosis) and damaged membranes, which prevent the expulsion of chemical substances from the body, culminating in scarring and necrosis.
Little is known about the cellular and molecular mechanisms underlying the alcoholic liver disease in different stages of liver tissue damage. Despite the progress made in several areas, an effective treatment to prevent this disease is still lacking. The Alcoholic Liver Disease (ALD) is seriously damaged, and no medicine with strong pertinence and reliable curative effect exists in clinic. According to different development stages and treatment purposes of alcoholic liver injury, the treatment approaches are as follows: abstinence from alcohol, nutritional therapy, pharmacotherapy and combination therapy. These treatments have certain limitations, for example, the abstinence of alcohol depends on the personal knowledge and self-control of the disease, and often fails due to the interference of other external factors; although the hormone treatment can relieve clinical symptoms, the death rate of serious diseases cannot be reduced, and the complications are more and difficult to popularize; some medicines have large toxic and side effects and bring liver burden, and medicines with strong pertinence and reliable curative effect are still lacked at present. Therefore, new signal paths need to be researched, and more basic research and clinical treatment targets are provided for drug treatment.
ROR α is one of the nuclear receptor superfamily and, at the same time, is also a lone nuclear receptor. Since they are widely distributed in various tissues in the body and have various important physiological functions, affecting the regulation of inflammatory response, the regulation of lipid metabolism, the development of nerve tissues, lymphocytes and the like, and the differentiation of B, T lymphocytes, there are some diseases related to the regulation of response, the regulation of lipid metabolism and the like, and there are attempts to use ROR α agonists, but there is no report on the study of SR1078 in the treatment of alcoholic liver injury.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide the application of SR1078 in preparing a medicament for treating alcoholic liver injury, wherein SR1078 can improve the condition of fatty accumulation liver injury.
Meanwhile, the invention also aims to provide a pharmaceutical composition for treating alcoholic liver injury.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
SR1078 application in preparing medicament for the treatment of alcoholic hepatic injury.
SR1078 is an agonist of retinoic acid receptor-related orphan receptor ROR α O, and the chemical structure of the agonist is shown as the following formula (1):
a pharmaceutical composition for treating alcoholic liver injury comprises SR1078 as active ingredient.
Optionally, the pharmaceutical composition further comprises pharmaceutically acceptable excipients.
Optionally, the pharmaceutical composition is in the form of injection, tablet or granule.
Further preferably, the pharmaceutical composition is prepared into an injection, the auxiliary materials comprise DMSO and physiological saline, the mass content of SR1078 in the injection is 0.1mg/ml, in the embodiment of the invention, the recommended usage amount of the injection is 10mg/kg/day, and the injection is injected intraperitoneally for 2 times a day.
The embodiment of the invention proves that: SR1078 improves the liver/body weight ratio of alcoholic liver disease mice; SR1078 can improve the serum enzymology index of alcoholic liver disease mouse, the mouse glutamic-pyruvic transaminase (ALT), glutamic-oxalacetic transaminase (AST) content is reduced obviously; SR1078 can improve endoplasmic reticulum stress condition of liver of alcoholic liver disease mouse, ROR alpha is raised, and key protein ATF4 and CHOP of endoplasmic reticulum stress are reduced; SR1078 can improve liver injury and lipid accumulation caused by alcohol. Provides a new strategy for treating alcoholic liver diseases, has important clinical treatment significance, shows that SR1078 can be used as an active ingredient of a medicament for treating alcoholic liver injury, and provides scientific basis for developing the medicament for treating alcoholic liver diseases in the future.
Drawings
FIG. 1 is a graph comparing the liver and body weight ratios of mice in experimental and control groups, wherein EtOH represents the control group and EtOH + SR1078 represents the experimental group;
FIG. 2 is a comparison graph of ALT and AST detection results of mice in an experimental group and a control group; wherein EtOH represents the control group and EtOH + SR1078 represents the experimental group;
FIG. 3 is a comparison graph of the results of the detection of key proteins of endoplasmic reticulum stress important in liver injury in mice of experimental group and control group; wherein EtOH is a control group, and SR1078 is an experimental group;
FIG. 4 is a graph comparing the results of HE staining and oil red O staining of liver histology of mice in experimental and control groups; wherein EtOH represents the control group and EtOH + SR1078 represents the experimental group.
Detailed Description
The invention will be further illustrated with reference to specific embodiments. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Furthermore, it should be understood that various changes and modifications can be made by those skilled in the art after reading the disclosure of the present invention, and equivalents fall within the scope of the appended claims.
Examples
This example demonstrates the therapeutic efficacy of ROR α agonist SR1078 on alcoholic liver disease:
1. materials:
c57BL/6 mice, male, 8-10 weeks old, purchased from Biotech limited, Wei Tony Hua, Beijing; SR1078 was purchased from Selleck, Shanghai; alcohol was purchased from alatin.
2. The method comprises the following steps:
2.1 Experimental groups and dosing
1) C57BL/6 male mice 8-10 weeks old are fed with common liquid feed to adapt to liquid feed for 5 days, then fed with lieber-Decarli alcohol feed (containing 5% alcohol) for 10 days, and fed with 31.5% alcohol (0.25g/ml) at 11 days according to 5g/kg/day dose, and the feed is divided into two groups, 10 mice in each group, one group is an experimental group, and the other group is a control group
Experimental groups SR1078 was administered while mice were gavaged with alcohol starting on day 11; the control group was given an equal volume of solvent (saline + DMSO (DMSO concentration less than 1%)) at the same time.
2) Administration dose: the experimental group was started on day 6 by gavage with 10mg/kg/day SR1078 twice daily (SR 1078 dissolved in DMSO to make stock solution concentration 100mg/ml, diluted to 0.1mg/ml with physiological saline just before use (final DMSO concentration less than 1%)), and the control group was given equal volume of solvent at the same time (physiological saline + DMSO (DMSO concentration less than 1%)).
3) The administration mode is as follows: the medicine is administrated by intraperitoneal injection.
4) The administration time is as follows: twice daily dosing was performed at 8:00 and 18:00 respectively.
2.2. Procedure of experiment
1) Body weight and liver detection: the weight of the mice after 10 days of administration was weighed using an electronic balance, and after the mice were anesthetized with isoflurane, the livers were removed, and the blood was blotted with absorbent paper and weighed. Washing with PBS for other experimental materials;
2) mouse serological detection: after each group of mice was anesthetized, whole blood was collected from the heart, coagulated at 4 ℃ for 8 hours, centrifuged at 3000rpm for 15min, and the supernatant was collected to prepare serum. Detecting the contents of alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) according to the kit instructions;
3) mouse liver molecular biology detection: detecting the expression conditions of ROR alpha, endoplasmic reticulum stress key protein ATF4 and CHOP protein in liver tissue by a protein imprinting technology: proteins were separated by SDS-PAGE gel and transferred to PVDF membrane, blocked with 5% milk, incubated overnight at 4 ℃ with ROR α, ATF4 and CHOP specific antibodies, washed 3 times with TBST and 1 hour with HRP-labeled secondary antibody, after washing 3 times with TBST, millipore's luminescence was added drop wise to the strip and developed with Amersham Imager 600, US GE ultrasensitive Multiplex Imager.
4) Mouse liver histology HE staining: after anesthetizing the mice, the livers were removed and cut into small pieces, and fixed by soaking in 4% paraformaldehyde. Before staining, the tissue blocks are washed by tap water overnight, then are subjected to gradient dehydration and permeabilization, are embedded by paraffin, and are sliced, spread, baked and baked. During dyeing, gradient hydration, hematoxylin dyeing, differentiation, eosin dyeing, dehydration, permeabilization and mounting are firstly carried out.
And taking a picture through a microscope;
5) male mouse liver histology oil red O staining: after anesthetizing the mice, the livers were removed and cut into small pieces, and fixed by soaking in 4% paraformaldehyde. After 48 hours, the cells were removed and dehydrated in 15%, 20% and 30% sucrose solutions. Embedding by an OCT embedding machine, and quickly freezing by liquid nitrogen. The slices were taken to-20 ℃ one day before slicing and were sliced at-20 ℃. During dyeing, the cells are washed by distilled water for 5min, soaked in 60% isopropanol for 2s, washed once by distilled water, the hematoxylin is used for counterstaining cell nuclei for 2s, the excessive water is absorbed by absorbent paper, and the cells are sealed by glycerol gelatin and photographed by a microscope.
3. The experimental results are as follows:
1) effect of SR1078 on mouse liver/body weight ratio:
respectively weighing the weights of the mice in the experimental group and the mice in the control group, and calculating the ratio of the liver to the weight of each group of mice, as shown in fig. 1, wherein the result shows that the ratio of the liver to the weight of the mice in the SR1078 treatment group (experimental group) is reduced, which indicates that SR1078 can relieve liver injury and reduce lipid accumulation, thereby reducing the ratio of the liver to the weight;
2) effect of SR1078 on serum enzymology indices of mouse alcoholic liver disease:
the ALT and AST contents in the blood serum of the mice of the experimental group and the control group are respectively detected, the results are shown in figure 2, and the experimental results show that the contents of alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) in the mice of the SR1078 treatment group (the experimental group) are obviously reduced compared with the control group;
3) SR1078 administration liver molecular biology detection:
as shown in fig. 3, the experimental results show that the ROR α of the SR1078 treatment group (experimental group) is increased, and the key proteins of endoplasmic reticulum stress, ATF4 and CHOP, are decreased, which indicates that the endoplasmic reticulum stress of the liver is decreased after the SR1078 treatment, indicating that SR1078 improves the lipid accumulation and damage of the liver of the mouse by relieving the endoplasmic reticulum stress;
4) SR1078 reduces liver damage and lipid accumulation:
as shown in the experimental results shown in fig. 4, HE staining results showed that the SR1078 treatment group (experimental group) suffered from liver injury alleviation; the results of oil red O staining showed a reduction in lipid accumulation in SR1078 treated group (experimental group).
The embodiment can show that: SR1078 improves the liver/body weight ratio of alcoholic liver disease mice; SR1078 can improve the serum enzymology index of alcoholic liver disease mouse; SR1078 can improve the endoplasmic reticulum stress situation of the alcoholic liver disease mouse liver; SR1078 can improve liver injury and lipid accumulation caused by alcohol, indicate SR1078 has the clinical application value applied to the treatment of alcoholic liver injury, can prepare the medicament for the treatment of alcoholic liver injury as the active ingredient.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (4)
- Application of SR1078 in preparing medicine for treating alcoholic liver injury.
- 2. A pharmaceutical composition for treating alcoholic liver injury, which comprises SR1078 as an active ingredient.
- 3. The pharmaceutical composition for the treatment of alcoholic liver injury of claim 2, further comprising a pharmaceutically acceptable adjuvant.
- 4. The pharmaceutical composition for treating alcoholic liver injury of claim 3, wherein the pharmaceutical composition is in the form of injection, tablet or granule.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113647359A (en) * | 2021-08-15 | 2021-11-16 | 芜湖职业技术学院 | Construction of a mouse model of acute liver injury induced by tunicamycin |
| CN117257919A (en) * | 2023-11-10 | 2023-12-22 | 南通大学附属医院 | Application of TBC1D15 in screening or preparing liver-protecting medicines |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113647359A (en) * | 2021-08-15 | 2021-11-16 | 芜湖职业技术学院 | Construction of a mouse model of acute liver injury induced by tunicamycin |
| CN117257919A (en) * | 2023-11-10 | 2023-12-22 | 南通大学附属医院 | Application of TBC1D15 in screening or preparing liver-protecting medicines |
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