CN115089580A - Application of thienopyridine medicine as medicine component in hepatitis medicine - Google Patents
Application of thienopyridine medicine as medicine component in hepatitis medicine Download PDFInfo
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K31/4353—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 ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4365—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 ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system having sulfur as a ring hetero atom, e.g. ticlopidine
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Abstract
The application of thienopyridine medicine as medicine component in preparing medicine for treating hepatitis. The present invention relates to a new pharmaceutical application. In the research, the invention discovers that (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate serving as a medicine component can reduce pathological damage of liver cells in autoimmune hepatitis and effectively relieve the autoimmune hepatitis. Further analyzing the action mechanism of the (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate in the clopidogrel hydrogen sulfate, finding that: the methyl (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) acetate is combined with a receptor P2Y12, and the biological activity of the P2Y12 receptor is inhibited, so that the generation of an inflammatory factor IFN-gamma in the liver is further reduced.
Description
Technical Field
The present invention relates to a new pharmaceutical application. In particular to the application of thienopyridine drugs (CAS: 113665-84-2) as a drug ingredient in hepatitis drugs, and more particularly to the application of (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyrido-5-yl) methyl acetate as a drug ingredient in hepatitis drugs.
Background
The liver, an important frontline immune organ, contains a large number of innate and adaptive immune cells that participate in a synergistic immune response. In autoimmune diseases, immune-mediated hepatitis occurs when hepatocytes are attacked by the body's immune system and T cells destroy infected hepatocytes in viral hepatitis. The underlying mechanisms of acute liver injury remain unclear, limiting the efficacy of clinical treatments. There is increasing evidence that activation of T cells plays an important role in promoting autoimmune hepatitis (AIH).
The medicine has the structural formula of Clopidogrel hydrogen sulfate (Clopidogrel Bisulfate for short), is a clinical medicine, has an inhibiting effect on platelet aggregation caused by adenosine diphosphate, and is mainly used for preventing and treating thrombotic events of cardiovascular diseases clinically.
At present, the new application of clopidogrel hydrogen sulfate in other diseases is not researched or disclosed.
Disclosure of Invention
The invention aims to find the application of the main component (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate in the clopidogrel hydrogen sulfate as a functional component in the autoimmune hepatitis medicament for the first time.
Structure of methyl (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothieno [3,2-c ] pyrid-5-yl) acetate
In the research, the invention discovers that (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate serving as a medicine component can reduce pathological damage of liver cells in autoimmune hepatitis and effectively relieve the autoimmune hepatitis. Further analyzing the action mechanism of the (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate in the clopidogrel hydrogen sulfate, finding that: the (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate is combined with a receptor P2Y12, and the biological activity of a P2Y12 receptor is inhibited, so that the generation of an inflammatory factor IFN-gamma in the liver is further reduced.
Autoimmune hepatitis (AIH) is an Autoimmune-reaction-mediated chronic progressive inflammatory disease of the liver characterized by hyperclobulinemia, autoantibody positivity, and histologically, hepatitis with plasma cell infiltration in the tract. In autoimmune diseases, immune-mediated hepatitis disease occurs when hepatocytes are attacked by the body's immune system. Concanavalin a (cona) is a phytohemagglutinin, which is protein in nature. ConA has strong mitogenic and lymphocyte transformation promoting effects. ConA is widely used in a model for simulating clinical immunological liver damage in animals because it causes damage to hepatocytes. The invention researches the application of (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate in the medicine for preventing the autoimmune hepatitis by using the autoimmune hepatitis induced by ConA as an animal model.
In an animal model of ConA-induced autoimmune hepatitis, the invention finds that (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] is compared with a control group]The clopidogrel hydrogen sulfate is added in the experiment to obviously relieve the liver pathological symptoms of the autoimmune hepatitis of mice, and the expression level of glutamic-oxaloacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT) which are relevant indexes of liver function examination of the autoimmune hepatitis is reduced. Also, IFN-. gamma.as a main causative agent of autoimmune hepatitis + The proportion of hepatic lymphocytes is obviously reduced. In addition, the secretion amount of proinflammatory factor IFN-gamma in peripheral blood is also obviously reduced. The comprehensive results show that (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3, 2-c)]And pyridine-5-yl) acetic acid methyl ester can effectively treat autoimmune hepatitis. The action mechanism is probably through reducing the pathogenicity IFN-gamma in autoimmune hepatitis + The hepatic lymphocyte ratio of (a). Further study of (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ]]The mechanism of action of methyl bipyridyl-5-yl) acetate (by addition of clopidogrel hydrogen sulfate) was found to be: the P2Y12 receptor is (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothiophene [3, 2-c)]And pyridine-5-yl) methyl acetate. The P2Y12 receptor isA G protein coupled receptor can promote the expression of inflammatory factors in lymphocytes to cause autoimmune diseases. Therefore, in autoimmune hepatitis, reduction of inflammatory factors produced by lymphocytes is beneficial to the alleviation of disease progression. (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothiophene [3, 2-c) in clopidogrel hydrogen sulfate]The methyl bipyridine-5-yl) acetate can reduce the activity of P2Y12 receptor by binding, reduce the generation of inflammatory factors and finally relieve the autoimmune hepatitis. In summary, the experimental data of the present invention indicate that (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] in clopidogrel hydrogen sulfate]The methyl bipyridyl-5-yl) acetate has a key effect in regulating and controlling the autoimmune hepatitis, and can provide an alternative medicine for treating the autoimmune hepatitis.
Drawings
FIG. 1: clinical clopidogrel bisulfate can effectively relieve ConA-induced autoimmune hepatitis
(A) The method comprises the following steps Molecular structure diagram of clopidogrel hydrogen sulfate.
(B) The method comprises the following steps Peripheral blood of five groups of different experimental mice is taken to detect the AST and ALT expression level.
(C) The method comprises the following steps Liver tissue sections from four different experimental rats were HE stained (D): statistical analysis of percent necrotic parts.
(E) The method comprises the following steps ELISA was used to detect the expression level of IFN-. gamma.in peripheral blood.
(F) The method comprises the following steps QPCR detects IFN-gamma mRNA expression in liver tissue;
*p<0.05，**p<0.01，***p<0.001(Student's t test)。
FIG. 2: clinical drug-clopidogrel hydrogen sulfate for preventively relieving ConA-induced autoimmune hepatitis
(A) The method comprises the following steps Flow detection analysis of NK in hepatic lymphocytes + IFN-γ + 、NKT + IFN-γ + 、CD4 + IFN-γ + 、CD8 + IFN-γ + Cell proportion
(B) The method comprises the following steps Percentage histogram.
*p<0.05，**p<0.01，***p<0.001(Student's t test)。
FIG. 3 therapeutic administration of clopidogrel hydrogen sulfate to alleviate autoimmune hepatitis (12h experiment)
(A) The method comprises the following steps Experimental arrangement pattern diagrams.
(B) The method comprises the following steps Peripheral blood of four groups of different experimental mice is taken to detect ALT and AST expression quantity. The grouping is as follows:
1. negative control;
2. positive control (+12.5mg/kg ConA), samples were taken after 12 h;
3. treatment group 2(+12.5mg/kg ConA), the gastric lavage inhibitor clopidogrel bisulfate (Clop) after 30min, and samples were taken after 12 h;
4. treatment group 4(+12.5mg/kg ConA), intragastric administration inhibitor clopidogrel bisulfate (Clop) after 1h, sampling after 12 h;
*p<0.05，(Student's t test)。
FIG. 4 therapeutic administration of clopidogrel hydrogen sulfate to alleviate autoimmune hepatitis (24h experiment)
(A) The method comprises the following steps Experimental arrangement pattern diagram.
(B) The method comprises the following steps Taking peripheral blood of three groups of different experimental mice, and detecting ALT and AST expression quantity. Wherein
1. Negative control;
WT + ConA. positive control (+12.5mg/kg ConA), sampled after 24 h;
3. treatment group (+12.5mg/kg ConA), gastric lavage inhibitor clopidogrel bisulfate (Clop) after 12h, sampling after 24 h;
*p<0.05(Student's t test)。
FIG. 5: knock-out P2Y12 receptor inhibits ConA-induced autoimmune hepatitis
(A) The method comprises the following steps Construction of an autoimmune hepatitis indicator map by ConA
(B) The method comprises the following steps Taking peripheral blood of four groups of different experimental mice, detecting AST and ALT expression level
(C) The method comprises the following steps HE staining of liver tissue sections of four different experimental mice
(D) The method comprises the following steps Detecting the expression quantity of IFN-gamma in peripheral blood by ELISA;
*p<0.05，**p<0.01(Student's t test)。
Detailed Description
Methyl 2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) acetate is the main component of clopidogrel hydrogen sulfate which is a clinical drug. The experiments in the following examples were carried out with addition of clopidogrel hydrogen sulfate as a source of methyl (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothieno [3,2-c ] pyrid-5-yl) acetate.
State of the art concanavalin a (cona) knowledge:
injection of ConA has been widely used to mimic the pattern of fulminant immune liver injury in vivo. In this model, administration of ConA rapidly activates T cells, such as CD4 + T cell, CD8 + T cells, natural killer T (nkt) cells, and Natural Killer (NK) cells, which then produce a variety of pro-inflammatory cytokines (e.g., tumor necrosis) factors (TNF- α, IFN- γ, IL-6) to cause hepatocyte injury by activating downstream signals. Therefore, the invention utilizes the autoimmune hepatitis induced by ConA to research (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3, 2-c)]The use of methyl bipyridin-5-yl) acetate in the prevention and treatment of autoimmune hepatitis and the molecular mechanisms provide valuable opportunities.
In this study, the present invention discusses the role of clinical methyl (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothieno [3,2-c ] pyrid-5-yl) acetate in ConA-induced autoimmune hepatitis. The invention discovers that methyl (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) acetate in clopidogrel hydrogen sulfate can effectively treat autoimmune hepatitis by means of both therapeutic administration and prophylactic administration. The invention discovers that (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate in clopidogrel hydrogen sulfate can reduce the proliferation and differentiation of T cells and the generation of inflammatory cytokines. Further action mechanism research finds that the action target of the methyl (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) acetate is related to the P2Y12 receptor.
Materials and methods
Animal(s) production
C57BL/6 mice were purchased from the Nanjing model animal center (Nanjing, China). The C57BL/6 mice used were 8-10 weeks old, housed in the SPF grade laboratory at the university of Hospital, free access to water and feed. P2Y12-KO mice were purchased from Jackson Laboratory. All experiments were approved and performed according to the guidelines of the animal protection committee of the college university.
AIH induction and treatment
C57BL/6 mice (8-10 weeks) were injected tail vein with 12.5mg/kg Con A (C2010, Sigma). And after 12h or 24h of tail vein injection, taking peripheral blood serum to carry out AST and ALT detection. And (3) drug treatment: a prevention and treatment group: clopidogrel bisulfate (Tinof Hangzhou pharmaceuticals Co., Ltd.) was dissolved in 0.9% NaCl and administered by gavage (clopidogrel bisulfate: 7.5mg/kg,15mg/kg,30mg/kg) at a dose 7 days before tail vein injection of ConA once a day until the end of the experiment, and 100. mu.L of 0.9% NaCl solution of the same volume was used as a control. ② administration treatment group: the mice were respectively administered (clopidogrel hydrogen sulfate: 15mg/kg) intragastrically 30min and 1h after tail vein ConA injection, and sampled 12h after modeling; mice were gavaged once 12h after tail vein ConA injection (clopidogrel bisulfate: 15mg/kg) and sampled 24h after modeling.
Histopathological and immunohistochemical analysis
Each mouse was deeply anesthetized with 200. mu.L of 10% chloral hydrate, then peripheral blood in each organ was removed by heart perfusion with 0.9% NaCl and fixed by 4% (w/v) paraformaldehyde perfusion. Liver tissue samples were fixed overnight at 4% paraformaldehyde and 4 ℃. After paraffin embedding, H & E staining analysis of inflammatory infiltration, and Image-Pro software for statistical analysis.
Cell staining and flow analysis
Liver lymphocytes were homogenized with a pre-cooled tissue homogenizer, collected by 70 μm cell strainer filtration into 15ml centrifuge tubes, centrifuged at 500g 4 ℃ for 10min to obtain cells, then resuspended in 8ml 33% Percoll, centrifuged at 2400g 20 ℃ density gradient for 25min, and cells at the bottom of 33% Percoll were collected, stained with CD4(Invitrogen,2210365), CD3(eBioscience,17-0031-82), CD8(eBioscience,11-0081-85), NK (BioLegend,108706), IFN-. gamma. (BioLegend,505810,1: 100). Flow analysis was a BD FACS Verse system and was performed using Flow Jo V10 software.
ELISA detection of inflammatory factor expression in peripheral blood
The WT, clopidogrel bisulfate treated group and P2Y12-KO mice were injected with tail vein ConA, respectively, and about 150. mu.l of serum was collected from orbital venous blood 12h or 24h after injection. Orbital venous blood was allowed to stand at room temperature for 30 minutes. After 30 minutes, serum was obtained by centrifugation at 4000rpm for 10 minutes at 4 ℃ and the supernatant was carefully collected. Clopidogrel bisulfate was dissolved in 0.9% NaCl and the AIH model was constructed by treating mice with (clopidogrel bisulfate: 7.5mg/kg,15mg/kg,30mg/kg) or solvent (0.9% NaCl). At 12h or 24h of modeling, approximately 150 μ l of serum was collected from orbital venous blood. Orbital venous blood was allowed to stand at room temperature for 30 minutes. After 30 minutes, serum was obtained by centrifuging at 4000rpm for 10 minutes at 4 ℃ and carefully collecting the supernatant without sucking down the pellet as much as possible. The concentration of IFN-. gamma.in each group of sera was measured using a specific ELISA kit (eBioscience) according to the manufacturer's instructions.
Statistical analysis
Data statistics were analyzed by Student's t-test and data are expressed as mean + -SEM, with P < 0.05 considered statistically significant.
As a result, the
3.1.1 clinical medication-clopidogrel hydrogen sulfate preventive medication for relieving autoimmune hepatitis
The invention evaluates whether the clopidogrel bisulfate has a certain treatment effect on the autoimmune hepatitis. Firstly, the invention purchases clinical medication clopidogrel hydrogen sulfate tablets from hospitals. The male mice, C57BL/6, were orally administered with different doses (clopidogrel hydrogen sulfate: 7.5mg/kg,15mg/kg,30mg/kg) for 7 days, and the control group was given 100. mu.L of 0.9% NaCl solution of the same volume as the control group. On day 7, a tail vein injection was performed with 12.5mg/kg Con A to construct an experimental autoimmune hepatitis model. Peripheral blood and liver tissue were taken 12h later for further experiments.
FIG. 1A is the molecular structural formula of clopidogrel hydrogen sulfate.
The results are shown in FIG. 1B, and show that the clopidogrel hydrogen sulfate 15mg/kg can remarkably relieve the severity of the disease and reduce the AST and ALT values. Taking clopidogrel bisulfate with the best AST and ALT reducing effect to perform liver HE staining.
As shown in fig. 1C, HE staining results indicated that the infiltration of leukocytes in the liver was significantly reduced and the necrotic area was smaller in the clopidogrel bisulfate-treated group compared to the control group.
Image-Pro software was statistically analyzed and statistically significant between the treated and control groups (fig. 1D).
Clopidogrel bisulfate at 15mg/kg simultaneously reduced the expression amount of IFN-. gamma.in peripheral blood (FIG. 1E), and IFN-. gamma.mRNA in liver tissue was also reduced (FIG. 1F).
In conclusion, research data indicate that the clinical medication, clopidogrel hydrogen sulfate, administered prophylactically relieves autoimmune hepatitis.
In order to fully prove the treatment effect of the clopidogrel hydrogen sulfate, the invention further detects IFN-gamma which is mainly used for causing the autoimmune hepatitis + The hepatic lymphocyte ratio of (1).
3.1.2 clinical medication-clopidogrel hydrogen sulfate preventive medication for relieving IFN-gamma positive proportion in autoimmune hepatitis
Different doses of clopidogrel bisulfate are intragastrically administered daily for 7 days before the AIH model is constructed. Then an AIH model is constructed, lymphocytes in the liver of the mouse are taken 12h later, and IFN-gamma related to disease activity infiltrated into the liver is detected through intracellular staining and flow analysis + Cell ratio. Flow analysis shows that the clopidogrel bisulfate treatment group has CD4 compared with a control group + T、CD8 + T, NK IFN-gamma in cells + The number of positives decreased overall as shown in fig. 2A and 2B. The data show that the positive proportion of IFN-gamma in autoimmune hepatitis is relieved by the clopidogrel bisulfate prophylactic administration.
The above results show that: the effective component (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate in the clopidogrel hydrogen sulfate medicine for preventive administration can effectively relieve the autoimmune hepatitis. To further investigate the therapeutic effect of methyl (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothieno [3,2-c ] pyrid-5-yl) acetate on experimental autoimmune hepatitis, therapeutic dosing was investigated.
3.1.3 clinical drug-clopidogrel hydrogen sulfate therapeutic drug delivery to alleviate autoimmune hepatitis
Firstly, an autoimmune hepatitis mouse model is constructed, oral administration and gastric lavage are carried out to clopidogrel hydrogen sulfate at different time points (30min and 1h) after ConA (12.5mg/kg) is injected into tail vein, and ALT and AST expression conditions in mouse peripheral blood serum are detected after 12 h.
Fig. 3A is a schematic diagram of an experimental schedule with clopidogrel bisulfate administration at various time points.
Fig. 3B shows the expression of ALT and AST at different time points.
The results show that: clopidogrel bisulfate at different time points can effectively reduce the expression conditions of ALT and AST. This further demonstrates that therapeutic administration of clopidogrel hydrogen sulfate, a clinical drug, can alleviate autoimmune hepatitis.
On the basis of therapeutic administration, the invention further prolongs the model time of autoimmune hepatitis, and researches the treatment effect of the effective component (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate in clopidogrel hydrogen sulfate on 24h experimental structural autoimmune hepatitis.
3.1.4 clinical drug-clopidogrel hydrogen sulfate therapeutic drug delivery for relieving autoimmune hepatitis
An autoimmune hepatitis mouse model is orally administrated, gavage is orally administrated, clopidogrel hydrogen sulfate is administrated 12h after ConA (12.5mg/kg) is injected into tail veins, and ALT and AST expression conditions in mouse peripheral blood serum are detected 24h later.
FIG. 4A is a schematic diagram of the experimental schedule for clopidogrel hydrogen sulfate administration 12h after injection of tail vein ConA (12.5 mg/kg).
FIG. 4B shows the expression of ALT and AST
The result is shown in figure 4, and the clopidogrel bisulfate after modeling for 12 hours can still effectively reduce the expression conditions of ALT and AST. This further demonstrates that therapeutic administration of clopidogrel hydrogen sulfate, a clinical drug, can alleviate autoimmune hepatitis.
In conclusion, the effective component (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate in the clopidogrel hydrogen sulfate can inhibit autoimmune hepatitis therapeutically, and the invention searches an action mechanism of clopidogrel hydrogen sulfate for treating the autoimmune hepatitis.
3.1.5 mechanism of action of clopidogrel bisulfate for relieving autoimmune hepatitis in clinical medication
The present invention first performed in vivo experiments using P2Y12-KO mice. The invention takes P2Y12-KO mice of 6-8 weeks to construct autoimmune hepatitis by means of tail vein injection ConA (as shown in figure 5A). AST and ALT were detected in peripheral blood after 12 h. The detection result shows that the values of AST and ALT of the P2Y12-KO + ConA mice are obviously lower than those of the WT + ConA mice. Further performing HE staining on the liver tissue, and finding out that: inflammatory cell infiltration in the liver was reduced in P2Y12-KO + ConA mice compared to WT + ConA group, as shown in fig. 5B and 5C. Recent studies have shown that Th1 cells secreting IFN- γ are the major pathological effector T cells of ConA-induced experimental autoimmune hepatitis. The invention detects the IFN-gamma content in the peripheral blood of two groups of mice, the result is shown in figure 5D, and the IFN-gamma expression quantity in the P2Y12-KO + ConA group is obviously lower than that in the WT + ConA group. In conclusion, the results are as follows: methyl (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothieno [3,2-c ] pyrido-5-yl) acetate from clopidogrel hydrogen sulfate may be a treatment for autoimmune hepatitis through P2Y12 receptor.
Discussion of the related Art
So far, the cause of autoimmune hepatitis is not clear, so that the cause of the autoimmune hepatitis is difficult to completely remove and the autoimmune hepatitis is difficult to cure. In clinic, hormone and immunosuppressant are mostly used to control the disease condition, control the immune response, stabilize the disease condition, keep the transaminase normal, and relieve or stabilize the disease condition, and many patients will relapse once the medicine is stopped.
The autoimmune hepatitis is mainly IFN-gamma + The research result of the invention shows that the preventive administration (clopidogrel hydrogen sulfate) can affect IFN-gamma of each cell subgroup + And (4) differentiation. A more interesting phenomenon is that clopidogrel bisulfate is dose-dependent. Wherein 15mg/kg ofClopidogrel bisulfate can effectively inhibit the expression level of AST and ALT. Therapeutic administration of clopidogrel bisulfate can also reduce the transaminase level of AIH mice and IFN-gamma of each cell subgroup + Differentiation, and pathological conditions of liver tissue. This further indicates that clopidogrel bisulfate can effectively inhibit ConA-induced autoimmune hepatitis. Further analysis of the mechanism of action of clopidogrel bisulfate: the research data of the invention show that the P2Y12 receptor may be (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2- c ]And a function target spot of the methylpyridine-5-yl) methyl acetate for treating experimental autoimmune hepatitis induced by ConA. These experimental results provide robust preliminary results for further studies in the future.
In conclusion, the invention proves that clinical medication-clopidogrel bisulfate can relieve clinical and histopathological conditions of autoimmune hepatitis by inhibiting the secretion of IFN-gamma in a liver lymphatic system in vivo, and the possible action mechanism is realized by a receptor P2Y 12. The theoretical basis is provided for the clinical index that (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate in clopidogrel hydrogen sulfate can effectively intervene in autoimmune hepatitis.
Claims (4)
1. An application of (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate as a medicine component in hepatitis medicine.
2. Use according to claim 1, wherein the pharmaceutical ingredient is provided by clopidogrel hydrogen sulfate.
3. The use of claim 1 or 2, wherein the data indicate that methyl (2S) -2- (2-chlorophenyl) -2- (4,5,6, 7-tetrahydrothieno [3,2-c ] pyrido-5-yl) acetate in clopidogrel hydrogen sulfate may be used for the treatment of autoimmune hepatitis via the P2Y12 receptor.
4. Use according to claim 3, characterized in that the mechanism of action of clopidogrel hydrogen sulfate is further analyzed: research data show that the P2Y12 receptor is probably the action target of (2S) -2- (2-chlorphenyl) -2- (4,5,6, 7-tetrahydrothiophene [3,2-c ] pyridine-5-yl) methyl acetate in clopidogrel hydrogen sulfate for treating ConA-induced autoimmune hepatitis.
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