CN104262462A - Glucose transporter inhibiting polypeptide as well as preparation method and application thereof - Google Patents
Glucose transporter inhibiting polypeptide as well as preparation method and application thereof Download PDFInfo
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- CN104262462A CN104262462A CN201410509098.3A CN201410509098A CN104262462A CN 104262462 A CN104262462 A CN 104262462A CN 201410509098 A CN201410509098 A CN 201410509098A CN 104262462 A CN104262462 A CN 104262462A
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Abstract
The invention provides a glucose transporter inhibiting polypeptide as well as a preparation method and application thereof, relating to the field of medicines. The invention in particular relates to a polypeptide compound which can be used for treating diabetes mellitus. The sequence of a polypeptide is MVTRCNNVGRYRVRIYVDA. The preparation method of the polypeptide is characterized in that the polypeptide is prepared by adopting the Fmoc protected solid phase synthesis technology; the polypeptide can be covalently linked with an adjuvant which is bovine serum albumin, human serum albumin or polyethylene glycol. The invention provides the application of the polypeptide to treatment of diabetes mellitus and complications of diabetes mellitus, wherein the complications of diabetes mellitus include diabetic nephropathy, diabetic hypertension, diabetic eye diseases and diabetic neuropathy; the polypeptide can be used for treating diabetes mellitus and complications of diabetes mellitus through various modes of administration, including hypodermic or intramuscular injection, intravenous injection or intravenous drip, administration of oral medicines, such as pills, capsules, and the like, nasal spray, and the like. The glucose transporter inhibiting polypeptide provided by the invention can inhibit glucose transporters in a targeted manner, thus achieving the effect of preventing or treating diabetes mellitus.
Description
Technical field:
The present invention relates to pharmaceutical field, be specifically related to the polypeptide compound being used for the treatment of diabetes.
Background technology
The metabolic disease of diabetes to be one group with hyperglycemia be feature is a kind of syndromes of glucose, protein and the lipid metabolism disorders caused by the cellular metabolism effect defect of insufficient insulin or Regular Insulin.In recent years, the number of diabetic subject increases gradually, is divided into type i diabetes and type ii diabetes.Wherein the number of type ii diabetes patient accounts for the 85-90% of diabetes total number of persons, and the health and lives of patient in serious threat.If diabetes can not get good control, a lot of complication may be caused, such as: diabetic nephropathy, retinopathy, hypertension etc.
At present, Most patients helps blood sugar in control volume by orally-taken blood sugar reducing medicine, or regulates insulin secretion etc., and a lot of patient finally still wants use of exogenous insulin.
The picked-up of cell to glucose is depended in the metabolism of glucose, but, glucose freely cannot enter cell by cell membrane lipid bilayer structure, and cell needs to be called for short glucose transporter (GLUT) transport function by the glucose transporter (glucose transporters) on cytolemma to the absorption of glucose and just can be achieved.
Suppress glucose transporter transport function just can suppress the absorption of glucose in body, alleviate the symptom of hyperglycemia in body, therefore, glucose transporter transhipment suppresses the target spot of thinking to treat diabetes.However, the glucose transporter transport inhibitors of unripe exploitation comes out, and is used for the treatment of diabetes.
Summary of the invention:
The present invention seeks to the feature for existing hypoglycemic medicine, design peptide species 2 compound, effectively can treat diabetes.
Technical scheme
A kind of glucose transporter suppresses polypeptide 2, and its sequence is MVTRCNNVGRYRVRIYVDA.The preparation method of described polypeptide 2, is characterized in that: the solid phase synthesis technique preparation adopting Fmoc protection, can a covalently bound adjuvant, and adjuvant is bovine serum albumin, human serum albumin, or polyoxyethylene glycol.The application of described polypeptide 2 in treatment diabetes and diabetic complication.Described diabetic complication is diabetic nephropathy, diabetic hypertension, diabetes eye illness, diabetic neuropathic pathology.Polypeptide 2, by multiple administering mode treatment diabetes and diabetic complication, comprises subcutaneous or intramuscular injection, intravenous injection or intravenous drip, oral administration as pill, capsule etc., nasal spray etc.
Beneficial outcomes:
Glucose transport body protein in the present invention suppresses polypeptide 2, can targeted inhibition glucose transport body protein, reaches the effect of prevention or treatment diabetes.Be more preferably the purposes in treatment diabetes and/or diabetes complicated disease drug.Described diabetes comprise type i diabetes and type ii diabetes; Described diabetic complication includes but not limited to: diabetic nephropathy, diabetic hypertension, diabetes eye illness, diabetic neuropathic pathology etc.
Embodiment
Embodiment 1
The chemical synthesis process of polypeptide
The solid phase synthesis technique preparation that polypeptide 2 is protected with Fmoc.Building-up reactions is carried out to N end from C end according to peptide sequence, and Rink medium (can buy in Advanced ChemTech company) has free amino group.In each step connection procedure, amino-acid residue all will activate, and has the HBTU of 4 times of free amino groups on medium in activator mixture, HOBt, DIEA and Fmoc-amino acid.After each amino acid whose ligation, all close the free amino group do not connected, capping 10 minutes with the mixture of a pyridine/acetic acid/N-Methylimidazole (4:1:0.5).After each amino acid whose ligation, next amino acid all will remove the Fmoc-group on medium before connecting, and goes Fmoc-group to use the dimethyl formamide containing 20% piperidines, needs 15 minutes.Finally, after all amino-acid residues are linked in sequence, polypeptide 2 98% trifluoroacetic acid cuts down from medium, and cutting at room temperature carries out 2 hours.
Apply above-mentioned electrochemical conditions can synthesize and obtain polypeptide 2, sequence is MVTRCNNVGRYRVRIYVDA, and this sequence is brand-new sequence.Also the raw work synthesis in Shanghai can be entrusted.
Embodiment 2
Glucose transport body protein suppresses the cell in vitro determination of activity of polypeptide 2
Adopt MTT colorimetry.By the insulinoma cell (INS-1 cell) of logarithmic growth, add in 96 well culture plates with 1.0 × 105, cultivate 24h, the Experimental agents glucose transport body protein that experimental port adds different concns respectively suppresses polypeptide 2 as experimental group; Blank group adds the solvent of same volume.Five multiple holes are established in every hole, and cultivate 48h, every hole adds MTT, after effect 4h, add DMSO, hatch 30min, absorbance A value is measured, by formula growth and proliferation of cell rate=(experimental group light absorption value/control group light absorption value-1) × 100% at microplate reader 570nm place.The EC50 calculating Experimental agents is 5.45nmol/L.
Embodiment 3
Glucose transport body protein suppresses hypoglycemic in the body of polypeptide 2 to be tested
Kunming mice, the high sugar of high fat is fed to body weight 18 ~ 22g, male and female half and half, and before experiment, water 24h is can't help in fasting, abdominal cavity disposable injection streptozotocin (STZ) 50mg/kg (citrate buffer of 1% dissolves preparation).After 1 week, blood sugar (BS) is surveyed in mouse tail vein blood sampling, and BS is modeling success higher than 16mmol/L.After modeling, mouse is divided into 3 groups, often organizes 10, diabetic model group (DM group): the PBS damping fluid subcutaneous injection giving equivalent; Control group (Bay g 5421): 20 μ g/kg subcutaneous administrations (being dissolved in 0.1mol/L PBS damping fluid), one day twice, continuous 15d; Sample sets (glucose transport body protein suppresses polypeptide 2): same to control group; Separately get 10 normal mouses as blank group.After medication, 3d, 9d, 15d are in tail venous blood sampling Quick Measurement blood sugar.After 15d, in drug withdrawal after 1 week, fasting be can't help water and is carried out glucose tolerance test in 16 hours.By 25% glucose solution (2g/kg) gavage, after gavage, 0,30,60,120 minute tail venous blood sampling detects blood sugar.
Result: glucose transporter protein polypeptide 2 has the effect (P<0.05, table 1) reducing diabetic mice blood sugar; Meanwhile, can the sugar tolerance (table 2) of diabetes-alleviating model mice.The effect of its polypeptide 2 is suitable with the effect of control group Bay g 5421.
Table 1 glucose transport body protein suppresses polypeptide 2 pairs of mouse hypoglycemic activities
* p<0.05, * * p<0.01 is compared with model group
Table 2 glucose transport body protein suppresses polypeptide 2 pairs of glucose tolerance in mice experimental results
* p<0.05, * * p<0.01 is compared with model group
Conclusion: glucose transport body protein suppresses polypeptide 2 pairs of diabetic mices to have therapeutic action.
SEQUENCE LISTING
Pu Luoda bio tech ltd, <110> Suzhou
<120> glucose transporter suppresses polypeptide and preparation method thereof, application
<130>
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 19
<212> PRT
<213> artificial sequence
<400> 1
Met Val Thr Arg Cys Asn Asn Val Gly Arg Tyr Arg Val Arg Ile Tyr
1 5 10 15
Val Asp Ala
Claims (6)
1. glucose transporter suppresses a polypeptide, it is characterized in that: its aminoacid sequence is MVTRCNNVGRYRVRIYVDA.
2. a preparation method for polypeptide described in claim 1, is characterized in that: the solid phase synthesis technique preparation adopting Fmoc protection.
3. the preparation method of polypeptide according to claim 2, is characterized in that: a covalently bound adjuvant, adjuvant is bovine serum albumin, human serum albumin, or polyoxyethylene glycol.
4. the application of polypeptide according to claim 1 in preparation treatment diabetes and diabetes complicated disease drug.
5. purposes according to claim 4, is characterized in that described diabetic complication is diabetic nephropathy, diabetic hypertension, diabetes eye illness, diabetic neuropathic pathology.
6. the purposes of polypeptide according to claim 4, is characterized in that: by multiple administering mode treatment diabetes and diabetic complication, comprise subcutaneous or intramuscular injection, intravenous injection or intravenous drip, oral administration as pill, capsule etc., nasal spray.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410509098.3A CN104262462B (en) | 2014-09-28 | Glucose transporter suppression polypeptide and preparation method thereof, application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410509098.3A CN104262462B (en) | 2014-09-28 | Glucose transporter suppression polypeptide and preparation method thereof, application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104262462A true CN104262462A (en) | 2015-01-07 |
| CN104262462B CN104262462B (en) | 2017-01-04 |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999059560A1 (en) * | 1998-05-15 | 1999-11-25 | Joslin Diabetes Center | Glucose transport modulators and uses therefor |
| CN102596247A (en) * | 2009-08-02 | 2012-07-18 | 两合生物科技公司 | Novel proteins |
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999059560A1 (en) * | 1998-05-15 | 1999-11-25 | Joslin Diabetes Center | Glucose transport modulators and uses therefor |
| CN102596247A (en) * | 2009-08-02 | 2012-07-18 | 两合生物科技公司 | Novel proteins |
Non-Patent Citations (2)
| Title |
|---|
| MOLITCH ME: "Current state of type 2 diabetes management.", 《AM J MANAG CARE.》 * |
| 宣吉明: "糖尿病治疗中多肽类药物新型缓控释给药制剂的研究进展", 《药学实践杂志》 * |
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Inventor after: Dai Renke Inventor after: Ding Haiyan Inventor after: Dai Tianming Inventor after: Guo Jiayin Inventor after: Jiang Weifan Inventor after: Zhang Chaojie Inventor after: Wang Zhenyu Inventor before: Luo Ruixue |
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Effective date of registration: 20161115 Address after: Guangji road 528400 Torch Development Zone in Guangdong province Zhongshan City No. 1 of Building 5 floor Applicant after: ZHONGSHAN PHARMASS Corp. Address before: Qingdao City, Shandong province 266033 Taiwan Liu Road No. 138 B Applicant before: Yang Gaolin Applicant before: GUANGDONG GAOHANG INTELLECTUAL PROPERTY OPERATION Co.,Ltd. Effective date of registration: 20161115 Address after: Tianhe District Tong East Road Guangzhou city Guangdong province 510665 B-101 No. 5, room B-118 Applicant after: GUANGDONG GAOHANG INTELLECTUAL PROPERTY OPERATION Co.,Ltd. Address before: Qingdao City, Shandong province 266033 Taiwan Liu Road No. 138 B Applicant before: Yang Gaolin |
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