CN110833620A

CN110833620A - Use of angiotensin converting enzyme inhibitors for the prevention and treatment of autoimmune diseases and the corresponding complications

Info

Publication number: CN110833620A
Application number: CN201810930382.6A
Authority: CN
Inventors: 刘密; 王镕
Original assignee: Individual
Current assignee: Individual
Priority date: 2018-08-15
Filing date: 2018-08-15
Publication date: 2020-02-25

Abstract

The invention provides the use of angiotensin converting enzyme inhibitors in the prevention and treatment of autoimmune diseases and the corresponding complications.

Description

Use of angiotensin converting enzyme inhibitors for the prevention and treatment of autoimmune diseases and the corresponding complications

Technical Field

The invention relates to the use of angiotensin converting enzyme inhibitors for the prevention and treatment of autoimmune diseases and the corresponding complications.

Background

With the increasing incidence of autoimmune diseases (autoimmune diseases), the public demand for new treatments for such diseases is increasing.

Angiotensin Converting Enzyme (ACE) is an enzyme associated with diseases such as hypertension and heart disease and is a target of many therapeutic drugs. Angiotensin Converting Enzyme (ACE) inhibitors have been approved for the treatment of hypertension and heart disease, etc., and the action mechanism is to reduce aldosterone secretion by inhibiting angiotensin converting enzyme, to reduce water and sodium retention, and to reduce venous return blood volume, which is beneficial to reducing heart preload. Meanwhile, the degradation of bradykinin is reduced, so that the blood vessel is expanded, the peripheral resistance is reduced, the load around the heart is reduced, and the cardiac output is increased. The pressure and the volume of the left ventricle at the end diastole are reduced, the tension of the ventricle wall is reduced, the resistance of the renal blood vessels is reduced, the renal blood flow is increased, and the improvement of the cardiac function is facilitated.

Autoimmune diseases refer to diseases caused by the body's immune reaction to autoantigens, which results in damage to the tissues. Autoimmune diseases can be divided into two main categories: organ-specific autoimmune diseases and systemic autoimmune diseases. The organ-specific autoimmune diseases mainly comprise insulin-dependent diabetes mellitus (type 1 diabetes), chronic lymphocytic thyroiditis, hyperthyroidism, myasthenia gravis, chronic ulcerative colitis, pernicious anemia accompanied by chronic atrophic gastritis, goodpasture syndrome, blain vulgaris, blain-like, primary biliary cirrhosis, multiple sclerosis, acute idiopathic polyneuritis and the like. Systemic autoimmune diseases are mainly Systemic Lupus Erythematosus (SLE), rheumatoid arthritis, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, autoimmune hemolytic anemia, thyroid autoimmune diseases, ulcerative colitis, and the like.

For example, in type 1 diabetes, Insulin (Insulin), glutamic acid decarboxylase 65KD isomer (GAD65), insulinoma binding protein-2 (IA-2), zinc transporter 8(ZnT8) and the like play a central role in the onset of type 1 diabetes, and it is the type 1 diabetes that is caused by the attack and killing of islet β cells by antigen-specific T cells caused by autoantigens such as Insulin.

Disclosure of Invention

The invention aims to provide a medicament which can prevent or/and treat autoimmune diseases, in particular, autoimmune diseases and corresponding complications can be prevented or/and treated by inhibiting the formation of any one or more of antigen epitopes on body autoantigens, namely chimeric polypeptides and natural polypeptides.

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

the invention provides an application of an angiotensin converting enzyme inhibitor (ACE inhibitor or ACEI) or a physiologically acceptable salt thereof in preparing a medicament for preventing or/and treating autoimmune diseases.

The use provided by the present invention is also characterized in that the autoimmune disease includes any one or more of type 1 diabetes, rheumatoid arthritis, chronic lymphocytic thyroiditis, hyperthyroidism, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, goodpasture's syndrome, pemphigus vulgaris, pemphigoid, primary biliary cirrhosis, multiple sclerosis, acute idiopathic polyneuritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, autoimmune hemolytic anemia, thyroid autoimmune disease, and ulcerative colitis.

The use provided by the invention is also characterized in that the autoimmune disease is type 1 diabetes.

The use according to the present invention is also characterised in that the angiotensin converting enzyme inhibitor is selected from ramipril, captopril, enalapril, benazepril, fosinopril, alacepril, lisinopril, perindopril, quinapril, delapril, cilazapril, spirapril, trandolapril, moexipril and imidapril or a pharmaceutically acceptable salt thereof.

The use provided by the invention is also characterized in that the angiotensin converting enzyme inhibitor is selected from ramipril or a pharmaceutically acceptable salt thereof.

The use provided by the invention is also characterized in that the medicament is in a form suitable for oral or parenteral administration.

The use provided by the invention is also characterized in that when the medicament is in a dosage form suitable for oral administration, the corresponding dosage form is any one of tablets, pills, capsules, powder, granules, syrup and ointment.

The use provided by the invention is also characterized in that the medicament is suitable for parenteral administration by injection. Parenteral administration includes, for example, intravenous, intramuscular, intraarterial, intraperitoneal, intranasal, intravesical (e.g., to the bladder), intradermal, topical, or subcutaneous administration. The corresponding preparation formulation is any one of powder injection, drop and injection.

The preferred route of administration is oral.

Action and Effect of the invention

According to the application of the angiotensin converting enzyme inhibitor or the physiologically acceptable salt thereof in preparing the medicament for preventing or/and treating the autoimmune disease, the angiotensin converting enzyme inhibitor can inhibit Angiotensin Converting Enzyme (ACE) and further inhibit the formation of chimeric antigen polypeptide (hybrid peptide), and the formation of the chimeric antigen polypeptide (hybrid peptide) is closely related to the activation of T cells and the attack β cells after the T cells are activated, so that the medicament prepared from the angiotensin converting enzyme inhibitor or the physiologically acceptable salt thereof can prevent and/or treat the autoimmune disease and corresponding complications by inhibiting the chimeric polypeptide of the antigen epitope of the autoantigen.

Drawings

FIG. 1 is a statistical chart of the results of the antigen-specific T cell activation assay performed in example 1;

FIG. 2 is a statistical chart of the experimental results of example 2; description of the drawings: 1: a control group; 2: 10 μ M ramipril treated group; 3: 2 μ M ramipril treated group.

FIG. 3 is a statistical chart of the experimental results of example 3.

Detailed Description

The present invention will be described below by taking an angiotensin converting enzyme inhibitor (ACE) as ramipril and taking an evaluation experiment of efficacy for the treatment of type 1 diabetes as an example. For the specific methods or materials used in the experiments, those skilled in the art can make routine alternatives according to the existing technologies based on the technical idea of the present invention, and not limited to the specific descriptions of the embodiments of the present invention.

The methods used in the experiments are conventional methods unless otherwise specified; the materials, reagents and the like used are commercially available unless otherwise specified.

Example 1

This example is to evaluate the efficacy of preventing and/or treating type 1 diabetes by inhibiting the formation of a chimeric polypeptide which is an epitope of the chimeric polypeptide on an angiotensin converting enzyme inhibitor body self-antigen.

In this embodiment, by preparing nanoparticles containing polypeptides, and allowing the nanoparticles to be phagocytized and released by antigen-presenting cells in a sample to be tested (in this embodiment, BDC6.9 spleen cells of a transgenic mouse), corresponding antigen-specific T cells associated with a disease can be activated, and by detecting a cell secretion secreted by the activated antigen-specific T cells, and based on a relationship between the cell secretion and the corresponding antigen-specific T cells, the content of the corresponding antigen-specific T cells can be determined as follows:

1. preparation of nanoparticles loaded with polypeptide

The polypeptides to be included in the experiment are chimeric polypeptides and polypeptides containing a partial sequence of the chimeric polypeptides, specifically three polypeptides, which are as follows:

chimeric polypeptide 1(hybrid peptide 1) having the polypeptide sequence: LQTLALNAARDP, antigen hybrid peptide 1 (epitope of antigen) associated with the onset of type 1 diabetes;

polypeptide 2(peptide 2) having the sequence:

PQVAQLELGGGPGAGDLQTLAL；

polypeptide 3(peptide 3), the polypeptide sequence being:

NAARDPNRDPNRESLDFLLV。

polypeptide 2 and polypeptide 3 are both polypeptides derived from related proteins of islet β cells, polypeptide 2 and polypeptide 3 are delivered simultaneously by nanoparticles so as to generate chimeric polypeptide 1(hybrid 1) after entering antigen presenting cells at a certain concentration, and the obtained nucleotide series of chimeric polypeptide 1 are derived from the underlined part in polypeptide 2 and the underlined part in polypeptide 3 respectively, so that chimeric polypeptide 1 is obtained.

The three polypeptides are respectively encapsulated and prepared into four types of nanoparticles according to four modes, specifically comprising the following steps: the nanoparticle coated with the chimeric polypeptide 1 (named as nanoparticle 1), the nanoparticle coated with the polypeptide 2 and the polypeptide 3 (named as nanoparticle 2 for convenience of description), the nanoparticle coated with the polypeptide 2 (named as nanoparticle 3 for convenience of description), and the nanoparticle coated with the polypeptide 3 (named as nanoparticle 4 for convenience of description).

The four kinds of nanoparticles are prepared by the following method:

step 1, dissolving the polypeptide to be entrapped into an aqueous solution, wherein the concentration content of the polypeptide is more than 0.1 mg/mL;

step 2, dissolving 200mg of polylactic-co-glycolic acid (PLGA) in 2ml of dichloromethane;

step 3, dripping the polypeptide aqueous solution obtained in the step 1 into dichloromethane containing PLGA, and carrying out ultrasonic treatment for more than 3 seconds;

step 4, adding the liquid obtained after the treatment in the step 3 into 5mL of 20mg/mL PVA aqueous solution, and carrying out ultrasonic treatment for more than 5 seconds;

step 5, adding the liquid obtained after the treatment in the step 4 into 100mL of 5mg/mL PVA solution, and stirring until the organic phase is completely volatilized;

step 6, centrifuging the liquid obtained by the step 5 for more than 7 minutes at the rotating speed of more than 8000RPM, removing the supernatant, resuspending the remaining precipitate in water, repeating the steps for many times, and finally obtaining the final precipitate with the supernatant removed;

step 7, suspending the final precipitate obtained in the step 5 in 20mL of 4% trehalose freeze-drying protective agent to obtain a suspension;

step 8, freeze-drying the suspension obtained in the step 7 to obtain freeze-dried powder, wherein the freeze-dried powder is used for a subsequent antigen-specific T cell activation experiment;

and 9, analyzing the powder obtained after freeze drying in the step 8, and determining the particle size of the nanoparticle coated with the polypeptide to be 100-500 nm.

2. Antigen specific T cell activation detection assay

The experiment was performed using BDC6.9 transgenic mouse spleen cells. The spleen is one of the main immune organs of the body and contains a large number of antigen presenting cells and antigen specific T cells related to immune reaction. The T Cell Receptor (TCR) on T cells in the spleen of BDC6.9 transgenic mice was engineered to recognize only specific antigens associated with the onset of type 1 diabetes, such as antigen 1, which contains an epitope of chimeric polypeptide 1 (LQTLALNAARDP). Therefore, only in the presence of this particular epitope can antigen-specific T cells be activated and recognize cells containing the relevant antigen. Upon stimulation with antigen 1, activated antigen-specific T cells secrete cytokines such as interferon-gamma.

Therefore, four antigen-specific T cell activation detection experiments are performed on the four nanoparticles prepared by the method, and the activation detection experiment samples corresponding to the

nanoparticles

1, 2, 3 and 4 are respectively sample 1, sample 2, sample 3 and sample 4, so that the nanoparticles are used for respectively presenting activated T cells, and the content of gamma interferon secreted by the activated antigen-specific T cells is detected to indirectly obtain the content of the activated T cells, so that whether the chimeric polypeptide 1 exists or not is known.

In addition, in the experiment, while the prepared nanoparticles respectively carrying various polypeptides are subjected to an antigen-specific T cell activation detection experiment, four antigen-specific T cell activation detection control samples are also respectively made in a free form of chimeric polypeptide 1, polypeptide 2, polypeptide 3 and polypeptide 2 and polypeptide 3 as control experiments, the four control samples are respectively named as control 1 (free chimeric polypeptide 1), control 2 (free polypeptide 2 and free polypeptide 3), control 3 (free polypeptide 2) and control 4 (free polypeptide 3), that is, the antigen-specific T cell activation detection experiment is not performed in the form of nanoparticles, but is performed directly in a free form.

In the experiment, an experiment for inhibiting antigen-specific T cell activation was also performed on samples of various nanoparticle groups and free polypeptide groups using an angiotensin converting enzyme inhibitor ramipril, the concentration of ramipril was 2 μ M, and the respective treatments of the nanoparticle combination free polypeptide groups were: the total eight inhibition group samples obtained by the method comprise nanoparticles coated with chimeric polypeptide 1, nanoparticles coated with polypeptide 2 and polypeptide 3, nanoparticles coated with polypeptide 2, nanoparticles coated with polypeptide 3, free polypeptide 1, free polypeptide 2, free polypeptide 3, free polypeptide 2 and free polypeptide 3, and are respectively named as inhibition 1, inhibition 2, inhibition 3, inhibition 4, inhibition 5, inhibition 6, inhibition 7 and inhibition 8.

In the antigen-specific T cell activation detection experiment, any one of the above-described antigen-specific T cell activation detection experiments is performed by using an enzyme-linked immunosorbent assay (ELISA) method to detect the content of γ interferon secreted from activated antigen-specific T cells in spleen cells. The specific steps for each activation detection assay were as follows:

step 1, separating and extracting mouse spleen cells from a mouse, and adding the mouse spleen cells into RPMI1640 culture medium containing 10% Fetal Bovine Serum (FBS), wherein the concentration of the spleen cells is more than 6000000 cells/mL;

step 2, preparation of antigen-specific T cell activation test sample: the nanoparticle or free polypeptide loaded with the polypeptide is suspended or dissolved in RPMI1640 medium containing 10% Fetal Bovine Serum (FBS) and mixed with mouse splenocytes suspended in RPMI1640 medium containing 10% FBS. The concentration of the mixed spleen cells is 6000000 cells/mL; in each group using nanoparticles, the final concentration of the nanoparticles containing the polypeptide obtained after mixing is 0.4 mg/mL; in each group using the free polypeptide, the final concentration of the free polypeptide obtained after mixing was 0.5. mu.g/mL. Then, ramipril was added to each of the inhibition groups at a final concentration of 2. mu.M. Thus, the sample 1, the sample 2, the sample 3, the sample 4, the control 1, the control 2, the control 3, the control 4, the inhibition 1, the inhibition 2, the inhibition 3, the inhibition 4, the inhibition 5, the inhibition 6, the inhibition 7 and the inhibition 8 to be subjected to the antigen-specific T cell activation detection experiment are obtained respectively;

step 3 Each group prepared in step 2 was placed in a cell incubator (37 ℃, 5% CO)₂) Culturing for three days;

after culturing for three days in the step 4, continuously centrifuging each group in the step 3 at 400g for 5 minutes, and collecting supernatant;

and 5, analyzing the content of the gamma interferon in the supernatant collected in each group obtained in the step 4 by adopting a double-antibody sandwich ELISA method.

3. Analysis of Experimental results

The analysis here is based on:

A. since the antigen-specific T cells in the spleen of BDC6.9 transgenic mice can recognize only chimeric polypeptide 1, free chimeric polypeptide 1 can directly stimulate and activate the secretion of cytokine gamma interferon by the antigen-specific T cells in the spleen cells, while the nanoparticles coated with chimeric polypeptide 1 can be phagocytized by antigen-presenting cells and release chimeric polypeptide 1 in the antigen-presenting cells, and the chimeric polypeptide 1 released in the antigen-presenting cells binds to MHC molecules in the antigen-presenting cells and is presented on the surface of the antigen-presenting cells. Thus, once chimeric polypeptide 1 is presented to the surface of an antigen presenting cell, it is recognized by antigen-specific T cells and stimulates secretion of interferon-gamma by antigen-specific T cells. By analyzing the content of interferon gamma by immunological analysis methods such as ELISA, it can be known how many antigen-specific T cells can recognize the corresponding antigen polypeptide and are activated.

B. And only the nanoparticles carrying the polypeptide 2 or only the nanoparticles carrying the polypeptide 3 release the entrapped polypeptide 2 or the polypeptide 3 after being phagocytized by antigen presenting cells, but the polypeptide 2 and the polypeptide 3 do not contain the complete sequence of the chimeric polypeptide 1(LQTLALNAARDP), so that no way is provided for activating antigen-specific T cells, and the antigen-specific T cells can not secrete gamma interferon.

C. Meanwhile, the nanoparticles carrying the polypeptide 2 and the polypeptide 3 release the polypeptide 2 and the polypeptide 3 simultaneously in the antigen presenting cells after being phagocytized by the antigen presenting cells. Because polypeptide 2 and polypeptide 3 each contain half the polypeptide sequence of chimeric polypeptide 1(hybrid polypeptide 1, LQTLALNAARDP), polypeptide 2 and polypeptide 3 can be cleaved by various enzymes in antigen presenting cells and rejoined together to form the complete chimeric polypeptide 1 (LQTLALNAARDP). Chimeric polypeptide 1(LQTLALNAARDP) formed by splicing polypeptide 2 and polypeptide 3 in antigen presenting cells is presented on the surface of antigen presenting cells after being combined with MHC molecules and activates antigen specific T cells to secrete gamma interferon.

FIG. 1 is a statistical chart of the results of the activation assay performed in example 1.

In fig. 1, 1: free chimeric polypeptide 1 treated group (control 1); 2: chimeric polypeptide 1 nanoparticle treated group (sample 1, corresponding to nanoparticle 1); 3: the treatment group carrying both polypeptide 2 and polypeptide 3 nanoparticles (sample 2, corresponding to nanoparticle 2); 4: only the polypeptide 2 nanoparticle treated group (sample 3, corresponding to nanoparticle 3) was loaded; 5: the group containing only polypeptide 3 nanoparticle treatment (sample 4, corresponding to nanoparticle 4); 6: group treated with free polypeptide 2 and free polypeptide 3 simultaneously (control 2); 7: free polypeptide 2 treated group (control 3); 8: free polypeptide 3 treated group (control 4); 9: free chimeric polypeptide 1 and 2 μ M ramipril treated group (inhibition 5); 10: chimeric polypeptide 1 nanoparticles and 2 μ M ramipril treated group (inhibition 1); 11: simultaneously carrying polypeptide 2 and polypeptide 3 nanoparticles and a 2 mu M ramipril treatment group (inhibition 2); 12: only carrying polypeptide 2 nanoparticles and 2 μ M ramipril treatment group (inhibition 3); 13: only containing polypeptide 3 nanoparticles and 2 μ M ramipril-treated group (inhibition 4); 14: free polypeptide 2 and free polypeptide 3 were treated simultaneously with 2 μ M ramipril (inhibition 6); 15: free polypeptide 2 and 2 μ M ramipril treated group (inhibition 7); 16: free polypeptide 3 and 2 μ M ramipril treated group (inhibition 8).

As shown in fig. 1, in

samples

3 and 4, the gamma interferon content was hardly detected, while in control 3 (free polypeptide 2) and control 4 (free polypeptide 3), the gamma interferon content was also hardly detected, excluding the possibility that polypeptide 2 and polypeptide 3, respectively, might be antigen-specific T cells.

In sample 1 (corresponding to nanoparticles loaded with chimeric polypeptide 1) and control 1 (corresponding to free chimeric polypeptide 1), the amount of interferon-gamma secreted by antigen-specific T cells after activation was the same. In sample 2 (corresponding to nanoparticles carrying both polypeptide 2 and polypeptide 3) carrying nanoparticles of two different polypeptides, antigen-specific T cells were also activated and secreted γ interferon, and the amount of γ interferon secreted by activated T cells was almost the same as in control 1 (containing free chimeric polypeptide 1), indicating that chimeric polypeptide 1 was produced in antigen-presenting cells and recognized by antigen-specific T cells. In contrast, in control 2, the interferon-gamma content was hardly detected, because although the group contained both free polypeptide 2 and free polypeptide 3, the free state could not be efficiently phagocytized by antigen-presenting cells and the chimeric polypeptide 1 was produced, which further indicates that the production of chimeric polypeptide 1 could be recognized by T cells.

In each of the treatment groups to which 2. mu.M ramipril was added, only the sample 1 and control 1 groups could detect the same amount of interferon gamma as in the absence of added ramipril. After addition of ramipril to sample 2, the gamma interferon levels were detected to be much lower than in the group without addition of ramipril. This also indicates that the production of chimeric polypeptide 1 is closely related to angiotensin converting enzyme. Ramipril is capable of inhibiting the production of chimeric polypeptide 1 in antigen presenting cells by inhibiting angiotensin converting enzyme and is capable of inhibiting the activation of antigen specific T cells by inhibiting the production of chimeric polypeptide 1.

In summary, the results of this example show that the antigen whose epitope is the chimeric polypeptide associated with pancreatic islet β can participate in the stimulation of the activation of antigen-specific T cells and thus the abnormal recognition of antigen-specific T cells to attack pancreatic islet β cells, and the formation of the chimeric polypeptide can be inhibited by the angiotensin converting enzyme inhibitor such as ramipril, so as to reduce the attack of antigen-specific T cells to pancreatic islet β cells, so that we can prevent and/or treat type 1 diabetes.

Example 2

The following is a description of example 2. In this embodiment, the same portions as those in embodiment 1 will be omitted.

This example is to evaluate the efficacy of angiotensin converting enzyme inhibitors in the prevention and treatment of type 1 diabetes by inhibiting epitopes that are natural polypeptides (native peptides) on the body's natural antigens.

Step 1, three groups are cultured respectively according to the following modes: mouse NIT-1 cells were cultured in RPMI1640 medium containing 10% FBS and high glucose (25mM glucose); one group was a control group, and two other groups were each supplemented with 2. mu.M and 10. mu.M ramipril as two inhibition groups. The cells were cultured continuously in a cell culture chamber (37 ℃ C., 5% CO2) for 7 days. The medium was changed daily and fresh 2. mu.M and 10. mu.M ramipril were added again in both inhibition groups.

Step 2, adding 8 mu M Thapsigargin (Thapsigargin) into each group of cells obtained in the step 1, and culturing in a cell culture box (37 ℃, 5% CO2) for 16 hours; during the culture period, fresh 2. mu.M or 10. mu.M ramipril was added to the ramipril-treated groups, respectively.

Step 3, washing each set of NIT-1 cells in step 2 by centrifugation to remove Thapsigargin. Fresh 2. mu.M and 10. mu.M ramipril were added simultaneously to both inhibition groups.

Step 4, NOD transgenic mouse spleen cells were collected and added to RPMI1640 medium containing 10% FBS and high glucose (25mM glucose). In both inhibition groups, 2. mu.M or 10. mu.M ramipril was added to the respective medium. Spleen cells were mixed with each group of NIT cells. The NIT cell concentration after mixing was 1000000 cells per ml and NOD mouse spleen cell concentration was 5000000 cells per ml. After mixing, each group of cells was cultured overnight in a cell culture chamber (37 ℃, 5% CO 2);

step 5, centrifuging each group of cells and collecting supernatant;

and 6, analyzing the content of the gamma interferon in the supernatant of each group collected in the step 5 by adopting an ELISA method.

The analysis basis of the results is as follows:

the spleen is one of main immune organs, such as antigen presenting cells and antigen specific T cells, so that spleen cells of the NOD mouse are adopted for culture in experiments.

NIT-1 cells are also β cells but have cancer cell characteristics and therefore can proliferate indefinitely when cultured in vitro NIT-1 cells can activate β cell antigen-specific T cells under stimulation of high glucose (25mM) and Thapsigargin by treating NIT-1 cells themselves to generate natural polypeptide (native peptides) antigens and chimeric polypeptide (hybrid peptides) antigens and can present the antigen polypeptides to the surface of NIT-1 cells through the MHC molecules of NIT-1 cells themselves, or by secreting insulin granules and the like to antigen presenting cells in mouse spleen cells, which can treat the proteins in the secretion granules of insulin granules and generate natural polypeptide (chimeric peptides) and chimeric polypeptide (chimeric peptides) antigens and can be detected by the antigen presenting cells once they are bound to the corresponding chimeric antigen presenting cell surface antigen of the chimeric peptide (chimeric peptide) 2 cells.

In this experiment, for the ramipril-inhibiting group, at least 7 days ahead, ramipril was added to the cell culture medium for pretreatment to inhibit the production of antigenic polypeptides in NIT-1 cells during the growth and division of NIT-1 cells, and the addition of ramipril during the culture after mixing NIT-1 cells with mouse spleen cells was to examine whether ramipril could effectively inhibit the production and presentation of antigenic polypeptides by NIT-1 cells through antigen-presenting cells in spleen cells and further activate β cell antigen-specific T cells.

FIG. 2 is a statistical chart of the experimental results of example 2.

In fig. 2, 1: a control group; 2: 10 μ M ramipril treated inhibition group; 3: 2 μ M ramipril treated inhibition group.

It can be seen from FIG. 2 that β cell-derived NIT-1 cells in the control group not treated with ramipril can activate β cell antigen-specific T cells in spleen cells of NOD mice after stimulation, however NIT-1 cells in the inhibitor group pretreated with ramipril cannot activate β cell antigen-specific T cells in spleen cells of NOD mice under stimulation, the antigen-specific T cells of mice have diversity, i.e., different antigen-specific T cell surfaces contain different T cell receptors, these different T cell surface T cell receptors can recognize both natural polypeptide epitopes and chimeric polypeptide epitopes, thus it can be seen that natural polypeptide (native peptides) epitopes or chimeric polypeptide (peptides) epitopes can activate antigen-specific T cells and enable antigen-specific T cell activation to attack β cells after activation of antigen-specific T cells, it is known from example 1 that ramipril can inhibit formation of chimeric polypeptide (ACE) by inhibiting Angiotensin Converting Enzyme (ACE) and thus it can be used for inhibiting the formation of only one antigen-specific polypeptide (peptide) in mice after activation of natural polypeptide and inhibition of chimeric polypeptide (peptide) by inhibiting the formation of mouse antigen-specific T cell.

Example 3

The following is a description of example 3. In this embodiment, the same portions as those in embodiment 1 will be omitted.

This example is to evaluate the effect of ramipril on oral inhibition of onset of type 1 diabetes in mice.

Newborn female NOD mice were used for this experiment. In the experiment, 10 NOD mice per group. The control group was not treated and was normally maintained. The ramipril treated group was orally fed with 5mg/kg/day ramipril per day from the second week. Mice blood glucose was recorded daily. The blood sugar is higher than 11.0 mmol.L^-1Onset of diabetes. And recording the diabetes onset of NOD mice at different time periods.

FIG. 3 is a statistical chart of the experimental results of example 3.

Results analysis NOD mice are type 1 diabetes model mice about 60% -85% of female NOD mice will develop type 1 diabetes after 20 weeks without intervention treatment from FIG. 3 it can be seen that female NOD mice have 70% diabetes after 22 weeks without intervention treatment but once we have given ramipril prophylactic treatment from NOD mice, only about 10% of mice will have type 1 diabetes after 22 weeks from the previous experimental results it is known that ramipril inhibits the production of antigenic polypeptides by inhibiting Angiotensin Converting Enzyme (ACE) and thus prevents antigen specific T cells from attacking β cells by recognizing antigenic polypeptides.

Examples effects and effects

According to the application of the angiotensin converting enzyme inhibitor or the physiologically acceptable salt thereof provided by the embodiment in preparing the medicament for preventing or/and treating the autoimmune diseases, because the angiotensin converting enzyme inhibitor can inhibit Angiotensin Converting Enzyme (ACE) and further inhibit the formation of the chimeric polypeptide antigenic polypeptide epitope, and the formation of the chimeric polypeptide antigenic polypeptide epitope is closely related to the activation of antigen-specific T cells and the attack β cells after the activation of the antigen-specific T cells, the medicament prepared from the angiotensin converting enzyme inhibitor or the physiologically acceptable salt thereof can prevent and/or treat type 1 diabetes and corresponding complications by inhibiting the formation of the chimeric polypeptide antigenic epitope of self antigens.

Further, since the angiotensin converting enzyme inhibitor can inhibit the formation of natural antigen polypeptides (hybrid peptides) by inhibiting Angiotensin Converting Enzyme (ACE), and the formation of natural antigen polypeptides (hybrid peptides) is also closely related to the activation of T cells and the attack β cells after the activation of T cells, the drug prepared from the angiotensin converting enzyme inhibitor or the physiologically acceptable salt thereof can also prevent and/or treat type 1 diabetes and the corresponding complications by inhibiting the formation of natural polypeptide epitopes of autoantigens;

further, the prevention and/or treatment of type 1 diabetes and the corresponding complications can be further provided by the chimeric polypeptide and the natural polypeptide which inhibit the epitope of the self-antigen at the same time.

In addition, in examples 1-3, the angiotensin converting enzyme inhibitor used is ramipril, and any angiotensin converting enzyme inhibitor capable of inhibiting angiotensin converting enzyme can inhibit the formation of chimeric polypeptide and/or natural polypeptide epitope of autoantigen related to autoimmune diseases by inhibiting angiotensin converting enzyme, so as to achieve the effect of preventing and/or treating autoimmune diseases and corresponding complications.

In examples 1 to 3, all of them are directed to type 1 diabetes, and in practice, if any one or both of the chimeric polypeptide and the natural polypeptide is present in the epitope of the autoantigen, the use of the angiotensin-converting enzyme inhibitor can achieve the effect of preventing and/or treating the autoimmune disease and the corresponding complications by inhibiting any one or more of the chimeric polypeptide and the natural polypeptide.

Claims

1. Use of an angiotensin converting enzyme inhibitor or a physiologically acceptable salt thereof for the preparation of a medicament for the prevention or/and treatment of autoimmune diseases and the corresponding complications.

2. Use according to claim 1, characterized in that:

wherein the autoimmune disease comprises any one or more of type 1 diabetes, rheumatoid arthritis, chronic lymphocytic thyroiditis, hyperthyroidism, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, goodpasture's syndrome, pemphigus vulgaris, pemphigoid, primary biliary cirrhosis, multiple sclerosis, acute idiopathic polyneuritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, autoimmune hemolytic anemia, thyroid autoimmune disease, and ulcerative colitis.

3. Use according to claim 2, characterized in that:

wherein the autoimmune disease is type 1 diabetes.

4. Use according to claim 1, characterized in that:

wherein the angiotensin converting enzyme inhibitor is selected from the group consisting of ramipril, captopril, enalapril, benazepril, fosinopril, alacepril, lisinopril, perindopril, quinapril, delapril, cilazapril, spirapril, trandolapril, moexipril and imidapril or a pharmaceutically acceptable salt thereof.

5. Use according to claim 1, characterized in that:

wherein the angiotensin converting enzyme inhibitor is selected from ramipril or a pharmaceutically acceptable salt thereof.

6. Use according to claim 1, characterized in that:

wherein the medicament is in a dosage form suitable for oral or parenteral administration.

7. Use according to claim 6, characterized in that:

when the medicament is in a dosage form suitable for oral administration, the corresponding dosage form is any one of tablets, pills, capsules, powder, medicinal granules, syrup and ointment.

8. Use according to claim 7, characterized in that:

wherein the medicament is administered parenterally in a form suitable for injection.

9. Use according to claim 8, characterized in that:

wherein, the corresponding preparation is any one of powder injection, drop and injection.