CN108330112A - Two B cell epitopes of HMG-CoA reductase and the Antigenic Peptide comprising wherein one or two epitope - Google Patents
Two B cell epitopes of HMG-CoA reductase and the Antigenic Peptide comprising wherein one or two epitope Download PDFInfo
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Abstract
The invention discloses two B cell epitopes of HMG COA reductases and include the Antigenic Peptide of wherein one or two epitope；This kind of Antigenic Peptide by HMG COA reductases one or two B epitope and insulinoma GAP-associated protein GAP (IA 2)) membrane-proximal region JM2 B epitope by connect peptide series connection, after series connection again by connect peptide be connected with the N-terminal of the T epitopes P2 of anti-diabetic.The diabetes model that multiple low dose STZ induces C57BL/6J mouse is injected by dorsal sc immunization route, has and significantly adjusts fat and hypoglycemic effect, while significantly reducing serum HMG COA reductases and uric acid level, improve the oxidation resistance of body.Antigenic Peptide provided by the invention can be used for recombinant microorganism and transgenic animals or plant, make them as production plant, it produces the Antigenic Peptide or makes oral vaccine, for diabetes, the treatment of tune fat, atherosclerosis, the relevant disease of oxidative stress such as diabetic nephropathy, senile dementia and gout.
Description
Technical field
The present invention relates to pharmacy and medicine related fields.More particularly to include one or both of HMG-CoA reductase B thin
The Antigenic Peptide of born of the same parents' epitope and its application in field of medicaments.
Background technology
Diabetes (Diabetes mellitus, DM) are the metabolic diseases characterized by insulin relative or absolute deficiency
Disease, insulin relative or absolute deficiency causes hyperglycemia, and then leads to three major nutrient metabolic disorder, and insulin absolutely lacks
For type 1 diabetes, insulin is opposite to be lacked for diabetes B.Type 1 diabetes (T1DM) are that the organ that is mediated by T cell is special
Property autoimmune disease, due to body autoreactivity immune response attack beta Cell of islet, so that it is gone to pot, afunction,
Absolutely lack caused diabetes so as to cause insulin.Induce body to causing autoimmunity by certain immunization route
Property disease autoantigen tolerance can effectively prevention of autoimmune diseases generation.The harm that diabetes are brought nearly all comes
From its complication.
Atherosclerosis is the major complications of diabetes, and it is solid that atherosclerosis is frequently accompanied by blood fat especially courage
The raising of alcohol, internal excessive cholesterol can cause the cardiovascular and cerebrovascular diseases such as atherosclerosis.HMG-CoA reductase (3- hydroxyls
Base -3- methyl glutaryl coenzyme A reductases, EC：1.1.1.34) it is rate-limiting enzyme during liver cell synthetic cholesterol, inhibits
HMG-CoA reductase can hinder endogenous cholesterol to synthesize.Cholesterol in serum level increase be angiocardiopathy it is high-risk because
Element, therefore the synthesis of cholesterol is hindered by inhibiting HMG-CoA reductase activity, blood cholesterol levels can be reduced to prevent
Atherosclerosis.A large amount of clinical research shows that oxidative stress plays an important role in diabetes and its complication, oxygen
Change stress refer to body when by various destructive stimulus, and activity in vivo oxygen radical generates excessive, and degree of oxidation is beyond oxidation
The Scavenging activity of product, oxidative system and antioxidant system are unbalance, so as to cause histiocytic damage.Research shows that diabetes
And its when vascular complication generation, oxidative stress obviously increases.Oxidative stress is also in diabetic nephropathy, senile dementia and pain
It plays an important role in wind.
P277 peptides are one section of specific polypeptides of 437~460 of HSP60, totally 24 amino acid, and can be with effect T
The antigenic determinant of cell effect is the specific fragment to play a role in T1DM.In order to increase the stability of peptide without changing
Its immune property, by original P277 peptides positioned at two cysteine C of the sequence the 6th and 11 by two knot propylhomoserin V come generation
It replaces, is named as DiaPep277, we have discovered that hypoglycemic effects of the DiaPep277 in treating model is better than P277.Have real
Research is tested to find, according to the identical subcutaneous administration modes of external DiaPep277, to be administered with P277 peptides, blood vessel endothelium can be caused
Cellular damage, and induce human body and generate antibody to induce serious atherosclerosis；By P277 be divided into P1 peptide fragments (437~
450) and P2 peptide fragments (448~460) two parts, discovery P1 polypeptides are the B epitope peptide fragment for inducing body and generating antibody, have and actuate
Pulse atherosclerosis acts on, and P2 is main anti-diabetic T epitope peptide fragments, does not induce atherosclerosis.
Proteantigen embodies its immunologic specificity by epitope.For a certain proteantigen, not only contain B cell table
Position, t helper cell epitope, cytotoxic T cell epitope, the knot closely related with Immune discrimination such as natural killer cells epitope
Structure, while also containing toxic epitope, the bad epitope such as cross reactivity epitope, these bad epitopes may cause Immune Deviation etc.
Adverse consequences, therefore in order to improve the protectiveness of proteantigen, need to make a choice in epitope levels, to obtain more preferably
Vaccine molecules.Have confirmed key effect of the P2 sections T epitopes peptide fragment in treating type 1 diabetes in P277, Wo Menyun
With B cell Antigen Epitope Prediction software and on-line prediction tool, to diabetic complication atherosclerosis GAP-associated protein GAP HMG-CoA
Reductase carries out B cell antigen epi-position analysis, and synthesis includes the antigen of one or two B cell epitopes of HMG-CoA reductase
Peptide.
Invention content
It is an object of the present invention to provide the linear B epitopes of HMG-CoA reductase, have SEQ ID No.1, SEQ ID
Amino acid sequence shown in No.2.
The second purpose of the present invention is to provide the N of the t helper cell epitope of HMG-CoA reductase B epitope and anti-diabetic
End, which passes through, connects the connected combined peptide of peptide, in the t helper cell epitope such as sequence table of anti-diabetic shown in SEQ ID No.3.
The three of the object of the invention are to provide the tandem polypeptide of three epitopes, the peptide by HMG-CoA reductase a B cell
Epitope is connected with the B epitope of insulinoma GAP-associated protein GAP (IA-2) membrane-proximal region JM2 by connecting peptide, then is passed through and connected peptide and anti-sugar
The N-terminal for urinating the t helper cell epitope of disease is connected in series, the B epitope such as sequence of insulinoma GAP-associated protein GAP (IA-2) membrane-proximal region JM2
Shown in table SEQ ID No.4.
The four of the object of the invention are to provide the tandem polypeptide of four epitopes, by two B cell epitopes of HMG-CoA reductase
It is connected with the B epitope of insulinoma GAP-associated protein GAP (IA-2) membrane-proximal region JM2 after series connection by connecting peptide, then by connecting peptide and resisting
The N-terminal of the t helper cell epitope of diabetes is connected in series.
Flexible joint wherein between B epitope is GG, SG, the flexible joint between B epitope and T epitopes be GGG, SGG or
Other suitable flexible peptides of person.
The five of the object of the invention are to provide comprising including HMG-CoA reductase B epitope described in immunological effective amount
The composition of Antigenic Peptide, also includes pharmaceutically acceptable carrier, and the immunological effective amount is every dose per kg body weight 2-5 millis
Gram.
The sixth object of the present invention is to provide the Antigenic Peptide for including one or two B epitope of HMG-CoA reductase
Prepare diabetes, adjust fat, atherosclerosis, the relevant disease of oxidative stress such as diabetic nephropathy, senile dementia and
Application in gout product.
The seventh object of the present invention is to provide the Antigenic Peptide for including one or two B epitope of HMG-CoA reductase
In prevention and treatment diabetes, adjust fat, atherosclerosis, the relevant disease of oxidative stress such as diabetic nephropathy, senile dementia
Application in disease and goat.
The beneficial effects of the present invention are：The invention discloses the B cell epitopes of HMG-CoA reductase, have SEQ ID
Amino acid sequence shown in No.1, SEQ ID No.2, ingredient is single, simple in structure, has broken away from bad epitope in native protein
It influences；The invention also discloses the Antigenic Peptides of a B cell epitope comprising HMG-CoA reductase, by HMG-CoA reductase B
Epitope is connected with the N-terminal of the t helper cell epitope of anti-diabetic by connecting peptide, which can reduce STZ and lure
The blood fat for the C57BL/6J mouse type 1 diabetes led is also disclosed comprising one or two B epitope in HMG-CoA reductase
Antigenic Peptide, the Antigenic Peptide by HMG-CoA reductase a B cell epitope and insulinoma GAP-associated protein GAP (IA-2) membrane-proximal region
The B epitope of JM2 is connected in series by connecting peptide series connection, then by the N-terminal of the t helper cell epitope of connection peptide and anti-diabetic,
Or by close with insulinoma GAP-associated protein GAP (IA-2) by connecting peptide after two B cell epitopes series connection of HMG-CoA reductase
The B epitope of film area JM2 is connected, then the N-terminal by connecting the t helper cell epitope of peptide and anti-diabetic is connected in series, both
Epitope peptide can reduce the C57BL/6J mouse type 1 diabetes blood glucose values of STZ inductions and adjust the blood fat of experimental animal, significantly drop
The content of HMG-CoA reductase in low serum, and serum uric acid level is significantly reduced, the oxidation resistance of body is improved, is had
There are important theory value and wide application prospect.
Description of the drawings
Fig. 1 ESI-MS measure molecular weight (Figure 1A of new type of peptides H243P, H441P, HIP1, H24IP1 of the present invention：H243P's
ESI-MS schemes；Figure 1B：The ESI-MS of H441P schemes；Fig. 1 C：The ESI-MS of HIP1 schemes；Fig. 1 D：The ESI-MS of H24IP1 schemes)
Mouse blood sugar tendency chart (Fig. 2A in Fig. 2 Experiment on therapy：P277 and DiaPep277 is small to the diabetes after STZ modelings
The comparison of blood glucose feelings trend in mouse Experiment on therapy；Fig. 2 B：HIP1, H24IP1 are to the diabetic mice Experiment on therapy after STZ modelings
In blood glucose value influence.)
Fig. 3 new type of peptides H243P, H441P, HIP1, H24IP1 is to blood fat in the diabetic mice Experiment on therapy after STZ modelings
Influence (Fig. 3 A：The influence of H243P, H441P pair the 7th week serum cholesterol；Fig. 3 B：H243P, H441P pairs of the 13rd week serum are sweet
The influence of oily three esters；Fig. 3 C：The influence of H243P pairs of the 7th week serum LDL cholesterol；Fig. 3 D：HIP1, H24IP1 couple
The influence of 8th week serum triglyceride；Fig. 3 E：The influence of HIP1, H24IP1 pair the 8th week serum LDL cholesterol)
The influence of Fig. 4 HIP1, H24IP1 to mice serum HMG-CoA reductase content
The influence of Fig. 5 HIP1, H24IP1 to mice serum uric acid level
Influence (Fig. 6 A of Fig. 6 HIP1, H24IP1 to diabetic mice Serum antioxidant indices：HIP1, H24IP1 are to peroxide
Changing hydrogenase activity (CAT) influences；Fig. 6 B：HIP1, H24IP1 are on glutathione peroxidase (GSH-PX) active influence)
Specific implementation mode
Using bioinformatics software and on-line analysis tool, with hydrophily, surface accessibility, antigenicity, flexibility
Based on, in conjunction with secondary structure prediction, filter out 2 linear B epitopes of HMG-CoA reductase, the t helper cell with anti-diabetic
The N-terminal of epitope by connect peptide combination, formed Antigenic Peptide H243P, H441P, and by the B cell epitope of HMG-CoA reductase with
The B epitope of insulinoma GAP-associated protein GAP (IA-2) membrane-proximal region JM2 is by connecting peptide series connection, then by connecting peptide and anti-diabetic
The N-terminal of t helper cell epitope is connected, and HIP1 is formed.Pass through connection after two B cell epitopes of HMG-CoA reductase are connected
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, and H24IP1 is formed.
The invention further relates to the compositions of the Antigenic Peptide comprising immunological effective amount.Antigenic Peptide immunology of the present invention has
Effect amount is usually every dose every kilogram of 2-5 milligrams, dosage can according to the age of patient, health status and individual reaction etc. into
One successive step；The Antigenic Peptide of safe and effective amount can be incomplete with pharmaceutically acceptable carrier such as Freund's complete adjuvant, Freund
The medicament forms such as vaccine are made in adjuvant, Lipofundin etc.；The Antigenic Peptide of safe and effective amount can also prepare various dosage forms, including speed
It releases or/and sustained release preparation；Pharmaceutical preparation can pass through any type and facilitate approach application, including subcutaneous, oral, flesh
Interior, mucous membrane, in peritonaeum or other parenteral or enteron aisle approach.Hereinafter reference will be made to the drawings, to the preferred of the present invention
Embodiment is described in detail.
Embodiment 1：The preparation process of new type of peptides H243P, H441P, HIP1, H24IP1
The B cell epitope sequences of HMG-CoA reductase are respectively EEEENKPNP and EPEIELPREPRPNE (see SEQ ID
No.1 and SEQ ID No.2)；The t helper cell epitope sequences of anti-diabetic are IPALDSLTPANED (see SEQ ID No.3)；
The B epitope sequence of insulinoma GAP-associated protein GAP (IA-2) membrane-proximal region JM2 is FEYQD (see SEQ ID No.4)；DiaPep277's
Amino acid sequence is VLGGGVALLRVIPALDSLTPANED (see SEQ NO.5)；Above-mentioned peptides are limited by gill biochemistry Shanghai
Company is synthesized using FMOC solid-phase synthesis, and HPLC detects purity ＞ 90%.
The specific building-up process of H243P, H441P, HIP1, H24IP1 is as follows：
1. selecting resin
For Peptide systhesis according to sequence inverse composition, polypeptide is connected since C-terminal is to N-terminal, Fmoc-Asp first (Otbu)-
OH Wang Resin；
2. removing Fmoc
Reactor is added in deprotection liquid (20% hexahydropyridine and 80%DMF), 30min or so is blown with nitrogen, then takes out
Fall, cleaned 5-6 times with DMF, then detect color of resin (generally in blue or brown).To remove before connecing next amino acid
Blocking group on a upper amino amino.
3. the condensation of amino acid
Reactor is added in ready raw material and solid condensing agent (tbtu, hobt) after deprotection, DMF then is added simultaneously
It is passed through nitrogen.Corresponding liquid activated dose (DIEA) is added after amino acid and condensing agent are completely dissolved to be reacted.Raw material with
Solid activator inventory is typically all three times.After reacting the regular hour, a small amount of resin is taken to detect, when resin is in water white transparency
When, show that the reaction was complete.
4. cutting
Synthetic polypeptide is added in cutting liquid, cutting liquid is filtered out after blender stirring 2-3h, is added wherein
Ether, polypeptide can be precipitated.Polypeptide is obtained by filtration in filter paper, and then washing 4-5 times with ether obtains crude product peptide.
5. peptide purification
It is purified after crude product peptide is drained, obtains the sterling of HPLC purity ＞ 90%.
ESI-MS detection synthesis peptide molecular weights.
Embodiment 2：The evaluating drug effect of H243P, H441P
1. the foundation of diabetic rat model
Streptozotocin (STZ) inducing mouse diabetes model is injected intraperitoneally using low dose of continuous several times.4 week old male
C57BL/6J mouse (are purchased from Yangzhou University's comparative medicine center, credit number：SCXK (Soviet Union) 2012-0004.) use 0.1M
PH4.4 citrate buffers configure STZ, are injected intraperitoneally by 40mg/kg dosage, one time a day, continuous injection 5 times.
The immunization therapy of 2.H243P, H441P are tested
It is hyperglycemia model that continuous two weeks after modeling, which measure blood glucose higher than 11.1mmol/l, by 50 type 1 diabetes moulds
Type mice group makes every group of blood glucose mean reach unanimity, respectively：Solvent control group (negative control group), P277 groups,
DiaPep277 groups, H243P groups, H441P groups.Administration group drug concentration is 1mg/ml, and subcutaneous administrations amount is 100 μ g/
Only/time, i.e. 100 μ l liquids；Negative control group：100 μ l finishes/only/time.Medicine ordinance method is dissolved in by 1mg+40mg mannitol
The proportional arrangement finish of 1ml20%Lipofundin.It is immunized 1 time weekly within continuous 5 weeks after Yu Chengmo, is immunized using dorsal sc
Mode, 0,1,2,3,4,6,8,10,12 week after mould, be immunized weekly primary.
Blood is taken through mouse intraocular corner of the eyes ball rear vein beard using capillary；Take blood 0.5mL or so every time, in 4000r/min from
Heart 10min centrifuges 2 separation serum, and obtained serum goes to -70 DEG C of refrigerators and preserve, be used for blood after -20 DEG C freeze one day
The measurement of clear indices.
Data are indicated with average value ± SD values, carry out statistical procedures with 17.0 softwares of SPSS, otherness compares between each group
Using independent samples t test.The * P ＜ 0.05 compared with solvent group, * * P ＜ 0.01；Compared with DiaPep277 groups, #P ＜
0.05, ##P ＜ 0.01；The , ＆P ＜ 0.05 compared with P277 groups.
3.P277 is compared with drug effect in DiaPep277 Experiment on therapy
Diabetic mice docking is taken into blood when measuring blood glucose, is used in combination blood sugar monitoring instrument detection mouse blood sugar horizontal, after Cheng Mo
4th week, the 7th week, the 8th week, DiaPep277 compared solvent significant difference (* * P ＜ 0.01), compared P277 significant differences within the 7th week
(＆P ＜ 0.05), the 9th week, DiaPep277 compared solvent significant difference (* P ＜ 0.05), shown in Fig. 2A.
The influence of 4.H243P, H441P to diabetic mice blood fat
With the detection of Beijing Northization safe clinical reagent Co., Ltd blood fat kit at solvent group after mould, DiaPep277 groups,
H243P groups, H441P groups the 7th week, the 13rd week mice serum triglycerides (TG), total cholesterol (TC), high-density lipoprotein courage are solid
The content of alcohol (HDL-C), low density lipoprotein cholesterol (LDL-C), the 7th week, H243P serum TC contents were substantially less than solvent group
(* * P ＜ 0.01), substantially less than DiaPep277 groups (#P ＜ 0.05), the 7th week, H441P serum TC contents were substantially less than solvent group
Shown in (* * P ＜ 0.01) Fig. 3 A；13rd week, H243P serum TG contents were substantially less than solvent group (* * P ＜ 0.01), substantially less than
DiaPep277 groups (##P ＜ 0.01), H441P serum TG contents are substantially less than solvent group (* * P ＜ 0.01), substantially less than
DiaPep277 groups (##P ＜ 0.01), shown in Fig. 3 B；7th week, H243P serum LDLs-C content was substantially less than solvent group (* * P ＜
0.01), substantially less than DiaPep277 groups (#P ＜ 0.05), shown in Fig. 3 C.
Embodiment 3：The pharmacodynamic evaluation of HIP1, H24IP1
1. the foundation of diabetic rat model
Streptozotocin (STZ) inducing mouse diabetes model is injected intraperitoneally using low dose of continuous several times.4 week old male
C57BL/6J mouse (are purchased from Yangzhou University's comparative medicine center, credit number：SCXK (Soviet Union) 2012-0004.) use 0.1M
PH4.4 citrate buffers configure STZ, are injected intraperitoneally by 40mg/kg dosage, one time a day, continuous injection 5 times.
The immunization therapy of 2.HIP1, H24IP1 are tested
It is hyperglycemia model that continuous two weeks after modeling, which measure blood glucose higher than 11.1mmol/l, by 40 type 1 diabetes moulds
Type mice group makes every group of blood glucose mean reach unanimity, respectively：Solvent control group (negative control group), DiaPep277 groups (sun
Property control group), HIP1 groups, H24IP1 groups.Administration group drug concentration is 1mg/ml, subcutaneous administrations amount be 100 μ g/ only/
It is secondary, i.e. 100 μ l liquids；Negative control group：100 μ l finishes/only/time.Medicine ordinance method is dissolved in by 1mg+40mg mannitol
The proportional arrangement finish of 1ml20%Lipofundin.It is immunized 1 time weekly within continuous 5 weeks after Yu Chengmo, is immunized using dorsal sc
Mode 0,1,2,3,4 week that is, after modeling, is immunized once weekly, and results of regular determination blood glucose value.
While measuring blood glucose, blood is taken through mouse intraocular corner of the eyes ball rear vein beard using capillary；Take blood 0.5mL left every time
The right side centrifuges 10min in 4000r/min, centrifuges 2 separation serum, and obtained serum goes to -70 DEG C after -20 DEG C freeze one day
Refrigerator preserves, and is used for the measurement of serum indices.
Data are indicated with average value ± SD values, carry out statistical procedures with 17.0 softwares of SPSS, otherness compares between each group
Using independent samples t test.The * P ＜ 0.05 compared with solvent group, * * P ＜ 0.01；Compared with DiaPep277 groups, #P ＜
0.05, ##P ＜ 0.01.
3. the influence pair blood glucose in diabetic mice trend
Diabetic mice docking is taken blood when measuring blood glucose, blood sugar monitoring instrument is used in combination to detect by continuous immunity 5 times after Yu Chengmo
Mouse blood sugar is horizontal.As a result as shown in Figure 2 B, after four administrations, HIP1 is substantially less than solvent group with H24IP1 group mouse blood sugars,
4th week, the 5th week, the 6th week, extremely significantly (* * P ＜ 0.01) compared to solvent difference, the 7th week, the 8th week compared to solvent significant difference (*
P ＜ 0.05), compare DiaPep277 differences extremely significantly (##P ＜ 0.01) within the 5th week, the 6th week, the 7th week.
4. the influence pair diabetic mice blood fat
With the detection of Beijing Northization safe clinical reagent Co., Ltd blood fat kit at solvent group after mould, DiaPep277 groups,
HIP1 groups, the 8th week mice serum triglycerides (TG) of H24IP1 groups, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-
C), the content of low density lipoprotein cholesterol (LDL-C), the 8th week, HIP1 was substantially less than solvent group with H24IP1 serum TG contents
(* P ＜ 0.05), while HIP1 is substantially less than DiaPep277 groups (#P ＜ 0.05), shown in Fig. 3 D；8th week H24IP1 group serum
LDL-C levels are substantially less than solvent group (* P ＜ 0.05), shown in Fig. 3 E.
5. the influence pair diabetic mice serum HMG-CoA reductase
With Nanjing Olympic blueness Bioisystech Co., Ltd produce HMG-CoA reductase ELISA kit detection solvent group,
The 5th week DiaPep277 groups, HIP1 groups, H24IP1 groups mice serum HMG-CoA reductase content, the results are shown in Figure 4, five times
The 5th week after administration, HIP1 serum HMG-CoA reductases compared with solvent group difference extremely significantly (* * P ＜ 0.01), H24IP1 with it is molten
Agent group significant difference (* P ＜ 0.05).
6. the influence pair diabetic mice serum uric acid
The uric acid water in uric acid detection kit the 8th week serum of detection of Bioengineering Research Institute's production is built up with Nanjing
It is flat.The results are shown in Figure 5, HIP1 and H24IP1 significant difference (the * P ＜ 0.05) compared with solvent group, compared with DiaPep277 groups
Significant difference (#P ＜ 0.05).
7. the influence pair diabetic mice catalase in serum, glutathione peroxidase
Catalase, the glutathione peroxidase detection kit of Bioengineering Research Institute's production are built up with Nanjing
Detect the 6th week serum oxidative stress index level, the results are shown in Figure 6, catalase in serum (CAT) content HIP1 with
H24IP1 has pole significant difference (* * P ＜ 0.01) compared to solvent.HIP1 has pole significant difference (##P ＜ compared to DiaPep277
0.01), there were significant differences (#P ＜ 0.05) compared to DiaPep277 by H24IP1.Glutathione peroxidase (GSH-PX)
H24IP1 has pole significant difference (* * P ＜ 0.01, ##P ＜ 0.01), HIP1 to have significantly compared to solvent compared to solvent and DiaPep277
Difference (* P ＜ 0.05).
Conclusion：Connection is passed through by the N-terminal of the t helper cell epitope of the B cell epitope and anti-diabetic of HMG-CoA reductase
The connected double epitope peptides of peptide can adjust the blood fat of experimental animal, by the B cell epitope and insulin of HMG-CoA reductase
The B epitope of tumor GAP-associated protein GAP (IA-2) membrane-proximal region JM2 is by connecting peptide series connection, then the T auxiliary by connecting peptide and anti-diabetic
Antigenic Peptide that the N-terminal of cell epitope is connected in series or by passing through company after two B cell epitopes series connection of HMG-CoA reductase
It connects peptide to connect 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 auxiliary
The Antigenic Peptide that the N-terminal of synergidae epitope is connected in series can reduce STZ induction C57BL/6J mouse type 1 diabetes blood glucose values and
The blood fat of experimental animal can be adjusted, while reducing the content of HMG-CoA reductase in serum, serum uric acid level is reduced, carries
The oxidation resistance of high body has important theory value and wide application prospect.
Finally illustrate, the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although passing through ginseng
According to certain examples of the present invention, invention has been described, but it will be apparent to an ordinarily skilled person in the art that can be with
Various changes are made to it in the form and details, without departing from of the invention defined by the appended claims
Spirit and scope.
Claims (10)
- 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. 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. 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. 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. 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. the Antigenic Peptide of the B cell epitope comprising HMG-CoA reductase described in the claim 1-4 comprising immunological effective amount Composition.
- 7. composition according to claim 6, it is characterised in that：It also include pharmaceutically acceptable carrier.
- 8. composition according to claim 6, it is characterised in that：Every dose per 2-5 milligrams of kg body weight.
- 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. 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.
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