CN108059656A - Micromolecule polypeptide and application thereof - Google Patents
Micromolecule polypeptide and application thereof Download PDFInfo
- Publication number
- CN108059656A CN108059656A CN201711476938.0A CN201711476938A CN108059656A CN 108059656 A CN108059656 A CN 108059656A CN 201711476938 A CN201711476938 A CN 201711476938A CN 108059656 A CN108059656 A CN 108059656A
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- Prior art keywords
- polypeptide
- micromolecule polypeptide
- micromolecule
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention discloses a kind of micromolecule polypeptides, and the amino acid sequence of micromolecule polypeptide is as shown in SEQ ID No.1.The present invention also provides the purposes of micromolecule polypeptide.The polypeptide of the present invention has good α glucosidase inhibitory actives and antioxidation activity, and stability is strong, and structure is easy to regulate and control, and has apparent application potential.
Description
Technical field
The present invention relates to a kind of micromolecule polypeptides.It is more particularly related to a kind of have alpha-glucosidase suppression
The micromolecule polypeptide of system activity and antioxidation activity.
Background technology
Diabetes are one group of metabolic diseases characterized by hyperglycaemia.Hyperglycaemia be then due to defect of insulin secretion or
Its biological effect is damaged, or both have concurrently and cause.Long-standing hyperglycaemia during diabetes, causes various tissues, particularly eye,
Kidney, heart, blood vessel, the chronic lesion of nerve, dysfunction.In the mechanism of action of current hypoglycemic medicine, by inhibiting to urge
Change the final step of starch or sucrose digestion and generate the alpha-glucosidase activity of glucide, it is considered to be control II type sugar
Urinate the desirable route of disease.The alpha-glucosidase restrainer clinically for treating diabetes clinically applied mainly has Ah card
Ripple sugar, voglibose, Miglitol etc., but these compounds have certain side effect.Therefore, it is necessary to prepare new α-
Glucosidase inhibitor.
The content of the invention
It is an object of the present invention to provide a kind of micromolecule polypeptides and application thereof, and the micromolecule polypeptide stability is strong, and
With with good alpha-glucosaccharase enzyme inhibition activity and antioxidation activity.
In order to realize these purposes and further advantage according to the present invention, a kind of micromolecule polypeptide is provided, small molecule is more
The amino acid sequence of peptide is as shown in SEQ ID No.1.
Preferably, the micromolecule polypeptide, the isoelectric point of the micromolecule polypeptide is 9.7.
The present invention also provides a kind of purposes of micromolecule polypeptide, are used to prepare the drug for treating diabetes.
The present invention also provides a kind of purposes of micromolecule polypeptide, are used to prepare the drug for removing free radical.
The present invention includes at least following advantageous effect：
1st, polypeptide of the invention has good alpha-glucosaccharase enzyme inhibition activity, is provided simultaneously with antioxidation activity.
2nd, the method for the present invention is a kind of micromolecule polypeptide, and stability is strong, and structure is easy to regulate and control, and there is apparent application to dive
Power.
3rd, micromolecule polypeptide of the present invention simple synthetic method compared with macromolecular polypeptides is easy to mass produce.
Part is illustrated to embody by further advantage, target and the feature of the present invention by following, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Description of the drawings
Fig. 1 is the free radical scavenging activity of polypeptide of the present invention and the relational graph of peptide concentration.
Specific embodiment
With reference to embodiment, the present invention is described in further detail, to make those skilled in the art with reference to specification
Word can be implemented according to this.
Embodiment 1
A kind of micromolecule polypeptide, the amino acid sequence of micromolecule polypeptide is as shown in SEQ ID No.1；
SEQ ID No.1：ICKKMMKKSTLLQDDIL.
The isoelectric point of the micromolecule polypeptide is 9.7.
The molecular weight of the micromolecule polypeptide is 2008.54g/mol.
The preparation method of the micromolecule polypeptide：
Amino acid-Wang resin is selected fully to be swollen resin with dichloromethane, with dimethyl formyl as carrier (resin)
Amine cleans several times, and with the DBLK of debita spissitudo, Fmoc- blocking groups are deviate from, is cleaned several times, washed with dimethylformamide afterwards
DBLK is removed, weighs suitable condensing agent benzotriazole-N, N, N', N'- tetramethylurea hexafluorophosphate and activator methyl
Second Fmoc- protected amino acid (Fmoc-Pro-OH) of quinoline and C-terminal is coupled, and is detected by ninhydrin detection method
Ensure that connection is more complete, cleaned several times with dimethylformamide, wash away remaining various residues, activator and condensing agent, according to
Amino acid sequence is coupled, and after all amino acid is connected, is sloughed last Fmoc- blocking groups, is used cutting liquid
Cracking removes resin and amino acid protective group, obtains the crude product of micromolecule polypeptide, and mass spectrum is sent to confirm molecular weight product
2008.54g/mol meet theoretical value.
Embodiment 2
The measure of the external alpha-glucosidase activity of micromolecule polypeptide.
All tests are all to use Microplate reader ELX808TM types microplate reader (BioTek companies of the U.S.), 37
It is measured under the conditions of DEG C.Data Analysis Software carries out data processing using Origin softwares, using acarbose as reference substance.
(1) preparation of inhibitor storing solution：The inhibitor tested be made into 10mM DMSO solution (1 polypeptide of embodiment
DMSO solution).
(2) preparation of enzyme stock solution：Alpha-glucosidase is purchased from Sigma Co., USA；With the phosphate-buffered of pH=6.8
Liquid is made into 1mg/mL respectively.
(3) preparation of Substrate stock liquid：P-nitrophenyl glucoside (PNPG) is substrate, purchased from Sigma companies；Use pH=
6.8 phosphate buffer be made into 10mg/mL respectively.
(4) preparation of terminate liquid：Sodium carbonate (Na2CO3) is purchased from Shanghai traditional Chinese medicines；With the phosphate buffer point of pH=6.8
0.1M Na are not made into2CO3Solution.
(5) test：The volume tested every time is all the phosphate buffer of the pH=6.8 of 200 μ L.
It is added in into 96 hole elisa Plates and is separately added into 10 μ L various concentrations inhibitor solutions (with pH=6.8 phosphate-buffereds
Solution dilution inhibitor storing solution), with pH=6.8 phosphate buffer solutions polishing to 170 μ L, then add in 10 μ L enzyme deposits
Liquid, keeps the temperature 10min in 37 DEG C of microplate reader, adds in 20 μ L Substrate stock liquid immediately, it is surveyed immediately in λ=405nm after mixing
Locate one minute absorbance change (slope).Reference liquid is pH=6.8 phosphate buffer solutions.
(6) result judges：Not to be loaded the absorbance change (slope) measured by product as 100 unit of activity；
Enzyme activity=(absorbance change (slope) of addition inhibitor/without the absorbance change of addition inhibitor
(slope) × 100,
When enzyme relative activity be 50 when inhibitor concentration be inhibitor IC50Value, the results are shown in Table 1：
1. polypeptide of table is to the IC of alpha-glucosaccharase enzyme inhibition activity50Value
| Drug | IC50(μM) |
| Polypeptide | 6.59 |
| Acarbose (reference substance) | 7.05 |
It can be seen that inhibitory activity (IC of the polypeptide to alpha-glucosidase of the present invention from the result of embodiment 250=6.59
μM) it is better than reference substance acarbose (IC50=7.05 μM).Experiment shows that the compound has strong inhibition alpha-glucosidase
Activity.The present invention provides new thinking to research and develop new antidiabetic medicine.
Embodiment 3
The DMSO solution (0.2,0.4,0.6,0.8 μ g/ml) of 1 polypeptide of 0.1ml embodiments is taken to be added to 8ml concentration be
In 0.004% 1,1- diphenyl -2- trinitrophenyl-hydrazines (DPPH) solution.(517nm) surveys absorbance at maximum wavelength, until
Until balance.Clearance rate calculation formula is as follows：
S%=(1-ASample)/ABlank× 100%.Wherein, ABlankNot add the absorbance of the DPPH solution of Peptide D MSO solution,
ASampleTo add in the absorbance of the DPPH solution of compound DMSO solution.The results are shown in Figure 1.As it can be seen that the peptide molecule of the present invention
With excellent antioxygenic property, more than 99.5% can be reached to the Scavenging activity of DPPH free radicals under low concentration.
Although the embodiments of the present invention have been disclosed as above, but its be not restricted in specification and embodiment it is listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, it is of the invention and unlimited
In specific details and shown here as the embodiment with description.
SEQUENCE LISTING
<110>Guangxi University of Chinese Medicine
<120>Micromolecule polypeptide and application thereof
<130> application
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 17
<212> PRT
<213>Polypeptide
<400> 1
Ile Cys Lys Lys Met Met Lys Lys Ser Thr Leu Leu Gln Asp Asp Ile
1 5 10 15
Leu
Claims (4)
1. a kind of micromolecule polypeptide, which is characterized in that the amino acid sequence of micromolecule polypeptide is as shown in SEQ ID No.1.
2. micromolecule polypeptide as described in claim 1, which is characterized in that the isoelectric point of the micromolecule polypeptide is 9.7.
3. a kind of purposes of micromolecule polypeptide as described in claim 1, which is characterized in that be used to prepare the medicine for the treatment of diabetes
Object.
4. a kind of purposes of micromolecule polypeptide as described in claim 1, which is characterized in that be used to prepare the medicine for removing free radical
Object.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711476938.0A CN108059656B (en) | 2017-12-29 | 2017-12-29 | Small molecule polypeptide and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711476938.0A CN108059656B (en) | 2017-12-29 | 2017-12-29 | Small molecule polypeptide and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108059656A true CN108059656A (en) | 2018-05-22 |
| CN108059656B CN108059656B (en) | 2020-07-17 |
Family
ID=62140791
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711476938.0A Active CN108059656B (en) | 2017-12-29 | 2017-12-29 | Small molecule polypeptide and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108059656B (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1603338A (en) * | 2004-08-24 | 2005-04-06 | 吉林大学 | Selenium-containing polypeptide and its use in medicine, food etc |
| CN104087643A (en) * | 2014-07-29 | 2014-10-08 | 哈尔滨伟平科技开发有限公司 | Method for preparing potato protein polypeptide |
| CN104187682A (en) * | 2014-09-25 | 2014-12-10 | 青岛嘉瑞生物技术有限公司 | Preparation technology for blood-sugar-reducing health Chinese herbal gluten peptides |
| CN104292309A (en) * | 2014-09-29 | 2015-01-21 | 广西中医药大学 | Micro-molecule polypeptide |
| CN104292307A (en) * | 2014-09-29 | 2015-01-21 | 广西中医药大学 | Polypeptide molecule |
| CN104327164A (en) * | 2014-09-29 | 2015-02-04 | 广西中医药大学 | Synthetic peptide and application thereof |
| CN104327165A (en) * | 2014-09-29 | 2015-02-04 | 广西中医药大学 | Polypeptide molecule and application thereof |
| CN104327168A (en) * | 2014-09-29 | 2015-02-04 | 广西中医药大学 | Micro-molecule polypeptide and application thereof |
-
2017
- 2017-12-29 CN CN201711476938.0A patent/CN108059656B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1603338A (en) * | 2004-08-24 | 2005-04-06 | 吉林大学 | Selenium-containing polypeptide and its use in medicine, food etc |
| CN104087643A (en) * | 2014-07-29 | 2014-10-08 | 哈尔滨伟平科技开发有限公司 | Method for preparing potato protein polypeptide |
| CN104187682A (en) * | 2014-09-25 | 2014-12-10 | 青岛嘉瑞生物技术有限公司 | Preparation technology for blood-sugar-reducing health Chinese herbal gluten peptides |
| CN104292309A (en) * | 2014-09-29 | 2015-01-21 | 广西中医药大学 | Micro-molecule polypeptide |
| CN104292307A (en) * | 2014-09-29 | 2015-01-21 | 广西中医药大学 | Polypeptide molecule |
| CN104327164A (en) * | 2014-09-29 | 2015-02-04 | 广西中医药大学 | Synthetic peptide and application thereof |
| CN104327165A (en) * | 2014-09-29 | 2015-02-04 | 广西中医药大学 | Polypeptide molecule and application thereof |
| CN104327168A (en) * | 2014-09-29 | 2015-02-04 | 广西中医药大学 | Micro-molecule polypeptide and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| ZAMBROWICZ ET AL.: "Multifunctional peptides derived from an egg yolk protein hydrolysate: isolation and characterization", 《AMINO ACIDS》 * |
| 励建荣 等: "蜂王浆水溶性蛋白质及其水解多肽对自由基的清除能力和对胰脂肪酶和α-葡萄糖苷酶的抑制活性", 《中国食品学报》 * |
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| Publication number | Publication date |
|---|---|
| CN108059656B (en) | 2020-07-17 |
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Effective date of registration: 20211105 Address after: 200120 room 205, 2f, No. 17, Lane 222, Guangdan Road, Pudong New Area, Shanghai Patentee after: Masala (Shanghai) Biotechnology Co.,Ltd. Address before: 530213 No. 13 Wuhe Avenue, Qingxiu District, Nanning City, Guangxi Zhuang Autonomous Region Patentee before: Guangxi University of Chinese Medicine |