CN108330112A - Two B cell epitopes of HMG-CoA reductase and the Antigenic Peptide comprising wherein one or two epitope - Google Patents

Abstract

The invention discloses two B cell epitopes of HMG-COA reductase and antigenic peptides containing one or two epitopes; this type of antigenic peptide is composed of one or two B-cell epitopes of HMG-COA reductase and insulinoma The B epitope of JM2 in the proximal membrane region of the associated protein (IA-2) is connected in series through a connecting peptide, and then connected to the N-terminus of the anti-diabetic T epitope P2 through a connecting peptide. The diabetes model of C57BL/6J mice was induced by multiple small doses of STZ injection through the back subcutaneous immunization route, which had significant lipid-lowering and blood-glucose-lowering effects, while significantly reducing serum HMG-COA reductase and uric acid levels, and improving the body's antioxidant capacity. The antigenic peptides provided by the present invention can also be used for recombinant microorganisms and transgenic animals or plants, making them as production plants to produce the antigenic peptides or make oral vaccines for diabetes, lipid-lowering, atherosclerosis, oxidative stress-related diseases Treatment of diseases such as diabetic nephropathy, Alzheimer's disease and gout.

Description

Two B-cell epitopes of HMG-CoA reductase and one or both of them antigenic peptide

technical field

The present invention relates to the fields of pharmacy and medicine. It particularly relates to an antigenic peptide containing one or two B cell epitopes in HMG-CoA reductase and its application in the field of medicine.

Background technique

Diabetes mellitus (DM) is a metabolic disease characterized by relative or absolute insufficiency of insulin. Relative or absolute insufficiency of insulin causes hyperglycemia, which in turn leads to metabolic disorders of the three major nutrients. Absolute insulin deficiency is type 1 diabetes, and relative insulin deficiency For type 2 diabetes. Type 1 diabetes mellitus (T1DM) is an organ-specific autoimmune disease mediated by T cells. The body's self-reactive immune response attacks the islet β cells, destroying them and losing their functions, leading to diabetes caused by absolute insulin deficiency. . Inducing the body's tolerance to autoantigens that cause autoimmune diseases through certain immune pathways can effectively prevent the occurrence of autoimmune diseases. Almost all the harm caused by diabetes comes from its complications.

Atherosclerosis is the main complication of diabetes. Atherosclerosis is often accompanied by an increase in blood lipids, especially cholesterol. Excessive cholesterol in the body can cause cardiovascular and cerebrovascular diseases such as atherosclerosis. HMG-CoA reductase (3-hydroxy-3-methylglutaryl coenzyme A reductase, EC: 1.1.1.34) is the rate-limiting enzyme in the synthesis of cholesterol in hepatocytes. Inhibition of HMG-CoA reductase can hinder endogenous Cholesterol synthesis. Elevated serum cholesterol level is a high risk factor for cardiovascular disease, so inhibiting the activity of HMG-CoA reductase to hinder the synthesis of cholesterol can reduce blood cholesterol level and prevent atherosclerosis. A large number of clinical studies have shown that oxidative stress plays an important role in diabetes and its complications. Oxidative stress refers to the excessive production of active oxygen free radicals in the body when the body is subjected to various harmful stimuli, and the degree of oxidation exceeds that of the oxidation products. The scavenging ability, oxidation system and anti-oxidation system are out of balance, which leads to the damage of tissue cells. Studies have shown that oxidative stress is significantly increased when diabetes and its vascular complications occur. Oxidative stress also plays an important role in diabetic nephropathy, Alzheimer's disease, and gout.

The P277 peptide is a specific polypeptide at positions 437-460 of HSP60, with a total of 24 amino acids. It is also an antigenic determinant that can react with effector T cells, and is a specific fragment that plays a role in T1DM. In order to increase the stability of the peptide without changing its immune properties, the two cysteine Cs at the 6th and 11th positions of the sequence in the original P277 peptide were replaced by two knottine Vs, named DiaPep277, we studied It was found that DiaPep277 had a better hypoglycemic effect than P277 in a therapeutic model. Existing experimental studies have found that according to the same subcutaneous administration method of foreign DiaPep277, administration of P277 peptide can cause damage to vascular endothelial cells, and induce the body to produce antibodies to induce severe atherosclerosis; P277 is divided into P1 peptides ( 437-450) and P2 peptide (448-460), it was found that the P1 polypeptide is a B epitope peptide that induces the body to produce antibodies, and has a pro-atherosclerotic effect, while P2 is the main anti-diabetic T epitope peptide segment, did not induce atherosclerosis.

Protein antigens reflect their immunospecificity through epitopes. As far as a protein antigen is concerned, it not only contains B cell epitopes, T helper cell epitopes, cytotoxic T cell epitopes, natural killer cell epitopes and other structures closely related to immune recognition, but also contains toxic epitopes, cross Undesirable epitopes such as reactive epitopes may cause adverse consequences such as immune bias. Therefore, in order to improve the protection of protein antigens, it is necessary to make selections at the epitope level to obtain more ideal vaccine molecules. It has been confirmed that the P2 segment T epitope peptide in P277 plays a key role in the treatment of type 1 diabetes. We used B cell epitope prediction software and online prediction tools to reduce the diabetic complication atherosclerosis-related protein HMG-CoA The enzyme performs B cell epitope analysis and synthesizes an antigenic peptide containing one or two B cell epitopes of HMG-CoA reductase.

Contents of the invention

One of the objects of the present invention is to provide the linear B epitope of HMG-CoA reductase, which has the amino acid sequence shown in SEQ ID No.1 and SEQ ID No.2.

The second object of the present invention is to provide a combined peptide in which the HMG-CoA reductase B epitope and the N-terminus of the anti-diabetic T helper cell epitope are linked by a connecting peptide, and the anti-diabetic T helper cell epitope is as shown in SEQ ID No in the sequence listing .3 shown.

The third object of the present invention is to provide a tandem peptide of three epitopes, which is composed of a B cell epitope of HMG-CoA reductase and the B epitope of the insulinoma-associated protein (IA-2) membrane-proximate region JM2 through the connecting peptide Tandem, and then formed by connecting the peptide and the N-terminus of the anti-diabetic T helper cell epitope, the B epitope of the near-membrane region of insulinoma-associated protein (IA-2) JM2 is shown in the sequence table SEQ ID No.4.

The fourth object of the present invention is to provide a tandem peptide of four epitopes, which is connected to the B epitope of the near-membrane region JM2 of the insulinoma-associated protein (IA-2) by connecting the two B cell epitopes of HMG-CoA reductase in series. The site is connected in series, and then connected in series with the N-terminus of the anti-diabetic T helper cell epitope.

Wherein the flexible linker between B epitopes is GG, SG, and the flexible linker between B epitope and T epitope is GGG, SGG or other suitable flexible peptides.

The fifth object of the present invention is to provide a composition comprising an immunologically effective amount of the antigenic peptide comprising the HMG-CoA reductase B epitope, and a pharmaceutically acceptable carrier, and the immunologically effective amount is 2-5 mg per kg body weight.

The sixth object of the present invention is to provide the antigen peptide containing one or two B epitopes of HMG-CoA reductase in the preparation of diabetes, lipid regulation, atherosclerosis, oxidative stress related diseases such as diabetic nephropathy, senile dementia disease and gout products.

The seventh object of the present invention is to provide the antigenic peptide containing one or two B epitopes of HMG-CoA reductase in the prevention and treatment of diabetes, lipid regulation, atherosclerosis, oxidative stress-related diseases such as diabetic nephropathy, Application in senile dementia and gout.

The beneficial effect of the present invention is that: the present invention discloses the B cell epitope of HMG-CoA reductase, has the amino acid sequence shown in SEQ ID No.1, SEQ ID No.2, and its composition is single, and structure is simple, gets rid of natural protein Influence of adverse epitopes; the present invention also discloses an antigenic peptide comprising a B cell epitope of HMG-CoA reductase, which is connected by the N-terminus of HMG-CoA reductase B epitope and anti-diabetic T helper cell epitope The two-epitope peptide can reduce the blood lipid of STZ-induced C57BL/6J mouse type 1 diabetes, and also discloses an antigen peptide containing one or two B epitopes on HMG-CoA reductase, the antigen The peptide consists of a B cell epitope of HMG-CoA reductase and the B epitope of the near-membrane region of insulinoma-associated protein (IA-2) JM2 in series through a connecting peptide, and then through connecting the peptide with an anti-diabetic T helper cell epitope The N-terminus is connected in series, or the two B cell epitopes of HMG-CoA reductase are connected in series, and the B epitope of the near-membrane region of insulinoma-associated protein (IA-2) JM2 is connected in series through a connecting peptide, and then connected to The N-terminal of the anti-diabetic T helper cell epitope is formed in tandem. These two epitope peptides can reduce the blood sugar level of STZ-induced C57BL/6J mice with type 1 diabetes and regulate the blood lipids of experimental animals, significantly reducing serum HMG-CoA Reductase content, and significantly reduce serum uric acid levels, improve the body's antioxidant capacity, has important theoretical value and broad application prospects.

Description of drawings

Fig. 1 ESI-MS measures the molecular weight of novel peptide H243P, H441P, HIP1, H24IP1 of the present invention (Fig. 1A: the ESI-MS figure of H243P; Fig. 1B: the ESI-MS figure of H441P; Fig. 1C: the ESI-MS figure of HIP1; Fig. 1D: ESI-MS image of H24IP1)

Figure 2 The blood glucose trend graph of mice in the treatment experiment (Figure 2A: Comparison of the blood sugar trend in the treatment experiment of diabetic mice after STZ modeling by P277 and DiaPep277; Figure 2B: The treatment of diabetic mice after STZ modeling by HIP1 and H24IP1 The influence of the blood sugar value in the experiment.)

Figure 3 Effects of novel peptides H243P, H441P, HIP1, and H24IP1 on blood lipids in diabetic mice after STZ modeling (Figure 3A: Effects of H243P and H441P on serum cholesterol at week 7; Effects of serum triglycerides at week 13; Figure 3C: Effects of H243P on serum LDL cholesterol at week 7; Figure 3D: Effects of HIP1 and H24IP1 on serum triglycerides at week 8; Figure 3E: Effects of HIP1 and H24IP1 on Effect of serum low-density lipoprotein cholesterol at week 8)

Figure 4 Effects of HIP1 and H24IP1 on mouse serum HMG-CoA reductase content

Figure 5 Effects of HIP1 and H24IP1 on serum uric acid levels in mice

Fig. 6 Effects of HIP1 and H24IP1 on serum antioxidant indexes of diabetic mice (Fig. 6A: effects of HIP1 and H24IP1 on catalase activity (CAT); Fig. 6B: effects of HIP1 and H24IP1 on glutathione peroxidase (GSH- PX) activity)

Detailed ways

Using bioinformatics software and online analysis tools, based on hydrophilicity, surface accessibility, antigenicity, and flexibility, combined with secondary structure prediction, two linear B epitopes of HMG-CoA reductase were screened out, The N-terminus of the T helper cell epitope of diabetes is combined by connecting peptides to form antigenic peptides H243P and H441P, and the B cell epitope of HMG-CoA reductase is combined with the B cell epitope of the insulinoma-associated protein (IA-2) proximal membrane region JM2 The epitopes are connected in series through a connecting peptide, and then connected in series with the N-terminus of the anti-diabetic T helper cell epitope to form HIP1. The two B cell epitopes of HMG-CoA reductase are connected in series with the B epitope of the near-membrane region of insulinoma-associated protein (IA-2) JM2 through a connecting peptide, and then connected with the anti-diabetic T helper cell surface through a connecting peptide. The N-termini of the bits are connected in series to form H24IP1.

The present invention also relates to a composition comprising an immunologically effective amount of said antigenic peptide. The immunologically effective dose of the antigenic peptide of the present invention is usually 2-5 milligrams per kilogram per dose, and the dosage can be further adjusted according to the age, health status and individual response of the patient; the safe and effective dose of the antigenic peptide can be compared with pharmaceutically acceptable Carriers such as Freund's complete adjuvant, Freund's incomplete adjuvant, Libaofening, etc. are made into pharmaceutical forms such as vaccines; safe and effective doses of antigenic peptides can also be prepared into various dosage forms, including immediate release or/and sustained release preparations; The pharmaceutical formulations can be administered by any convenient route, including subcutaneous, oral, intramuscular, mucosal, intraperitoneal or other parenteral or enteral routes. Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

Example 1: Preparation process of novel peptides H243P, H441P, HIP1, H24IP1

The B cell epitope sequences of HMG-CoA reductase are EEEEENKPNP and EPEIELPREPRPNE (see SEQ ID No.1 and SEQ ID No.2); the antidiabetic T helper cell epitope sequence is IPALDSLTPANED (see SEQ ID No.3); The B epitope sequence of the membrane-proximal region of insulinoma-associated protein (IA-2) JM2 is FEYQD (see SEQ ID No.4); the amino acid sequence of DiaPep277 is VLGGGVALLRVIPALDSLTPANED (see SEQ NO.5); the above peptides were produced by Jill Biochemical Shanghai Co., Ltd. adopts FMOC solid-phase synthesis method to synthesize, and the purity detected by HPLC is more than 90%.

The specific synthesis process of H243P, H441P, HIP1, H24IP1 is as follows:

1. Choose resin

The peptide synthesis is reversely synthesized according to the sequence, and the peptides are connected from the C-terminus to the N-terminus. First, Fmoc-Asp(Otbu)-OH Wang Resin;

2. Remove Fmoc

Add the deprotection solution (20% hexahydropyridine and 80% DMF) into the reactor, blow it with nitrogen for about 30 minutes, then remove it, wash it with DMF 5-6 times, and then check the color of the resin (usually blue or brown). Before taking the next amino acid, the protecting group on the amino group of the previous amino acid is removed.

3. Condensation of amino acids

After deprotection, the prepared raw materials and solid condensing agent (tbtu, hobt) are added to the reactor, and then DMF is added and nitrogen gas is introduced. After the amino acid and the condensing agent are completely dissolved, the corresponding liquid activator (DIEA) is added for reaction. The amount of raw material and solid activator is generally tripled. After reacting for a certain period of time, take a small amount of resin for detection, and when the resin is colorless and transparent, it indicates that the reaction is complete.

4. Cutting

Add the synthesized polypeptide into the cutting solution, stir with a stirrer for 2-3 hours, filter out the cutting solution, add ether to it, and the polypeptide can be precipitated. Filter paper to obtain the peptide, and then wash with ether 4-5 times to obtain the crude peptide.

5. Peptide purification

The crude peptide was extracted to dryness and then purified to obtain a pure product with an HPLC purity > 90%.

The molecular weight of synthetic peptides was detected by ESI-MS.

Example 2: Drug efficacy evaluation of H243P and H441P

1. Establishment of diabetic model mice

A mouse model of diabetes was induced by continuous intraperitoneal injection of streptozotocin (STZ) in small doses. 4-week-old male C57BL/6J mice (purchased from the Comparative Medicine Center of Yangzhou University, license number: SCXK (Su) 2012-0004.) were configured with STZ in 0.1MpH4.4 citrate buffer, and intraperitoneal at a dose of 40mg/kg Injection, once a day, 5 consecutive injections.

2. Immunotherapy experiment of H243P and H441P

After two consecutive weeks of modeling, the blood glucose was measured to be higher than 11.1mmol/l as a hyperglycemia model, and 50 type 1 diabetes model mice were divided into groups to make the average blood glucose values of each group tend to be consistent, respectively: solvent control group (negative control group) , P277 group, DiaPep277 group, H243P group, H441P group. The drug concentration in the administration group was 1 mg/ml, and the subcutaneous injection dose was 100 μg/animal/time, that is, 100 μl liquid medicine; the negative control group: 100 μl oil/animal/time. Drug preparation method The oil preparation is prepared according to the ratio of 1mg+40mg mannitol dissolved in 1ml 20% Lipofundin. Immunize once a week for 5 consecutive weeks after modeling, adopt the way of subcutaneous immunization on the back, that is, immunize once a week after 0, 1, 2, 3, 4, 6, 8, 10, and 12 weeks after modeling.

Use a capillary tube to collect blood through the venous plexus behind the inner canthus of the mouse eye; take about 0.5mL of blood each time, centrifuge at 4000r/min for 10min, and centrifuge twice to separate the serum. Store in a -70°C refrigerator for the determination of various serum indicators.

The data were expressed as the mean ± SD value, and SPSS 17.0 software was used for statistical processing, and the differences among the groups were compared using the independent sample t test. Compared with solvent group, *P<0.05, **P<0.01; compared with DiaPep277 group, #P<0.05, ##P<0.01; compared with P277 group, &P<0.05.

3. Comparison of efficacy between P277 and DiaPep277 in treatment experiment

When measuring blood sugar, blood was taken from the tail of diabetic mice, and the blood sugar level of the mice was detected with a blood glucose monitor. At the 4th, 7th, and 8th weeks after modeling, there was a significant difference between DiaPep277 and the solvent (**P<0.01) , the difference was significant compared with P277 at week 7 (&P<0.05), and the difference between DiaPep277 and solvent was significant at week 9 (*P<0.05), as shown in Figure 2A.

4. Effects of H243P and H441P on blood lipids in diabetic mice

The serum triglyceride (TG), total cholesterol (TC), The content of high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C), at week 7, the serum TC content of H243P was significantly lower than that of the solvent group (**P<0.01), significantly lower than that of the DiaPep277 group (#P<0.05), at the 7th week, the serum TC content of H441P was significantly lower than that of the solvent group (**P<0.01) as shown in Figure 3A; at the 13th week, the serum TG content of H243P was significantly lower than that of the solvent group (**P<0.01) 0.01), significantly lower than the DiaPep277 group (##P<0.01), H441P serum TG content was significantly lower than the solvent group (**P<0.01), significantly lower than the DiaPep277 group (##P<0.01), as shown in Figure 3B ; At week 7, the serum LDL-C content of H243P was significantly lower than that of the solvent group (**P<0.01), and significantly lower than that of the DiaPep277 group (#P<0.05), as shown in Figure 3C.

Example 3: Pharmacodynamic evaluation of HIP1 and H24IP1

1. Establishment of diabetic model mice

A mouse model of diabetes was induced by continuous intraperitoneal injection of streptozotocin (STZ) in small doses. 4-week-old male C57BL/6J mice (purchased from the Comparative Medicine Center of Yangzhou University, license number: SCXK (Su) 2012-0004.) were configured with STZ in 0.1MpH4.4 citrate buffer, and intraperitoneal at a dose of 40mg/kg Injection, once a day, 5 consecutive injections.

2. Immunotherapy experiment of HIP1 and H24IP1

After two consecutive weeks of modeling, the blood glucose was measured to be higher than 11.1mmol/l as a hyperglycemia model, and 40 type 1 diabetes model mice were divided into groups to make the average blood glucose values of each group tend to be consistent, respectively: solvent control group (negative control group) , DiaPep277 group (positive control group), HIP1 group, H24IP1 group. The drug concentration in the administration group was 1 mg/ml, and the subcutaneous injection dose was 100 μg/animal/time, that is, 100 μl liquid medicine; the negative control group: 100 μl oil/animal/time. Drug preparation method The oil preparation is prepared according to the ratio of 1mg+40mg mannitol dissolved in 1ml 20% Lipofundin. Immunize once a week for 5 consecutive weeks after modeling, adopt the way of subcutaneous immunization on the back, that is, immunize once a week after 0, 1, 2, 3, and 4 weeks after modeling, and measure blood sugar regularly.

While measuring blood sugar, blood was collected through the venous plexus behind the inner canthus of the mouse eye using a capillary tube; about 0.5 mL of blood was collected each time, centrifuged at 4000 r/min for 10 min, and centrifuged twice to separate serum, and the obtained serum was frozen at -20 °C One day later, transfer to -70°C refrigerator for storage, and use for determination of serum indexes.

The data were expressed as the mean ± SD value, and SPSS 17.0 software was used for statistical processing, and the differences among the groups were compared using the independent sample t test. Compared with the solvent group, *P<0.05, **P<0.01; compared with the DiaPep277 group, #P<0.05, ##P<0.01.

3. Effects on blood glucose trends in diabetic mice

Immunization was continued for 5 times after the model was formed, and the blood was collected from the tail of the diabetic mice when the blood glucose was measured, and the blood glucose level of the mice was detected with a blood glucose monitor. The results are shown in Figure 2B. After four administrations, the blood glucose of the mice in the HIP1 and H24IP1 groups was significantly lower than that in the solvent group, and the difference between the 4th week, the 5th week, and the 6th week was extremely significant compared to the solvent group (**P<0.01 ), there was a significant difference between the 7th week and the 8th week compared with the solvent (*P<0.05), and the difference between the 5th week, the 6th week, and the 7th week was extremely significant compared with DiaPep277 (##P<0.01).

4. Effects on blood lipids in diabetic mice

Blood lipid kits from Beijing Beihua Kangtai Clinical Reagent Co., Ltd. were used to detect serum triglyceride (TG), total cholesterol (TC), and high-density lipoprotein in mice in solvent group, DiaPep277 group, HIP1 group, and H24IP1 group at week 8 after modeling Cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) content, at the 8th week, the serum TG content of HIP1 and H24IP1 was significantly lower than that of the solvent group (*P<0.05), and HIP1 was significantly lower than that of the DiaPep277 group ( #P<0.05), as shown in Figure 3D; the level of serum LDL-C in the H24IP1 group at week 8 was significantly lower than that of the vehicle group (*P<0.05), as shown in Figure 3E.

5. Effect on serum HMG-CoA reductase in diabetic mice

The HMG-CoA reductase ELISA kit produced by Nanjing Aoqing Biotechnology Co., Ltd. was used to detect the serum HMG-CoA reductase content of mice in the solvent group, DiaPep277 group, HIP1 group, and H24IP1 group at the fifth week. The results are shown in Figure 4. At week 5 after five administrations, there was a significant difference in serum HMG-CoA reductase between HIP1 and the vehicle group (**P<0.01), and a significant difference between H24IP1 and the vehicle group (*P<0.05).

6. Effect on serum uric acid content in diabetic mice

The uric acid level in the serum at week 8 was detected with the uric acid detection kit produced by Nanjing Jiancheng Bioengineering Institute. The results are shown in Figure 5, HIP1 and H24IP1 were significantly different from the solvent group (*P<0.05), and significantly different from the DiaPep277 group (#P<0.05).

7. Effects on serum catalase and glutathione peroxidase in diabetic mice

The catalase and glutathione peroxidase detection kits produced by Nanjing Jiancheng Bioengineering Institute were used to detect the level of serum oxidative stress indicators in the sixth week. The results are shown in Figure 6. Serum catalase (CAT ) content of HIP1 and H24IP1 were significantly different (**P<0.01). HIP1 has a very significant difference (##P<0.01) compared with DiaPep277, and H24IP1 has a significant difference (#P<0.05) compared with DiaPep277. Glutathione peroxidase (GSH-PX) H24IP1 is significantly different from solvent and DiaPep277 (**P<0.01, ##P<0.01), HIP1 is significantly different from solvent (*P<0.05) .

Conclusion: The double-epitope peptide connected by the B-cell epitope of HMG-CoA reductase and the N-terminus of the anti-diabetic T helper cell epitope can regulate the blood lipids of experimental animals, and a B-cell epitope of HMG-CoA reductase Cell epitope and the B epitope of insulinoma-associated protein (IA-2) near the membrane region JM2 are connected in series through the connecting peptide, and then the antigenic peptide formed by connecting the peptide in series with the N-terminal of the anti-diabetic T helper cell epitope, or The two B cell epitopes of HMG-CoA reductase are connected in series, and then the B epitope of the near-membrane region of insulinoma-associated protein (IA-2) JM2 is connected in series through the connecting peptide, and then the anti-diabetic T helper cell surface is connected through the connecting peptide. The antigenic peptide formed by the N-terminus of the N-terminus in series can reduce the blood sugar level of STZ-induced C57BL/6J mice with type 1 diabetes and can regulate the blood lipids of experimental animals, while reducing the content of HMG-CoA reductase in serum and serum uric acid level. Improving the antioxidant capacity of the body has important theoretical value and broad application prospects.

Finally, it is noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described with reference to some examples of the present invention, those of ordinary skill in the art should understand that it can be described in the form Various changes may be made in matter and details thereof without departing from the spirit and scope of the invention as defined in the appended claims.

Claims (10)

1. The B cell epitope of 1.HMG-CoA reductases and the Antigenic Peptide comprising wherein one or two epitope, the HMG-CoA is also The B cell epitope of protoenzyme has amino acid sequence shown in SEQ ID No.1 and SEQ ID No.2.
2. 2. Antigenic Peptide according to claim 1, auxiliary by a B cell epitope of HMG-CoA reductase and the T of anti-diabetic The N-terminal of synergidae epitope is composed by connecting peptide, SEQ ID in the t helper cell epitope of anti-diabetic such as sequence table Shown in No.3.
3. 3. Antigenic Peptide according to claim 1, related to insulinoma by a B cell epitope of HMG-CoA reductase The B epitope of albumen (IA-2) membrane-proximal region JM2 is by connecting peptide series connection, then the t helper cell table by connecting peptide and anti-diabetic The N-terminal of position is connected in series, the B epitope such as sequence table SEQ ID No.4 institutes of insulinoma GAP-associated protein GAP (IA-2) membrane-proximal region JM2 Show.
4. 4. Antigenic Peptide according to claim 1, by passing through connection after two B cell epitopes series connection of HMG-CoA reductase Peptide is connected with the B epitope of insulinoma GAP-associated protein GAP (IA-2) membrane-proximal region JM2, then the T by connecting peptide and anti-diabetic is assisted The N-terminal of cell epitope is connected in series.
5. 5. the Antigenic Peptide of the B cell epitope comprising HMG-CoA reductase according to claim 2-4, it is characterised in that：Institute It is GG, SG to state the flexible joint between B epitope, and the flexible joint between B epitope and T epitopes is GGG, SGG or other are suitable Flexible peptide.
6. 6. the Antigenic Peptide of the B cell epitope comprising HMG-CoA reductase described in the claim 1-4 comprising immunological effective amount Composition.
7. 7. composition according to claim 6, it is characterised in that：It also include pharmaceutically acceptable carrier.
8. 8. composition according to claim 6, it is characterised in that：Every dose per 2-5 milligrams of kg body weight.
9. 9. the B cell epitope of the HMG-CoA reductase according to claim 1-4 and including wherein one or two epitope Antigenic Peptide preparing diabetes, adjusting fat, atherosclerosis, the relevant disease of oxidative stress such as diabetic nephropathy, old silly Application in slow-witted disease and gout product.
10. 10. the B cell epitope of the HMG-CoA reductase according to claim 1-4 and including wherein one or two table Position Antigenic Peptide prophylactic treatment diabetes, adjust fat, atherosclerosis, the relevant disease of oxidative stress such as diabetic nephropathy, Application in senile dementia and gout.