CN108101963A - A kind of oligose peptide and preparation method thereof - Google Patents
A kind of oligose peptide and preparation method thereof Download PDFInfo
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- CN108101963A CN108101963A CN201810090447.0A CN201810090447A CN108101963A CN 108101963 A CN108101963 A CN 108101963A CN 201810090447 A CN201810090447 A CN 201810090447A CN 108101963 A CN108101963 A CN 108101963A
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- peptide
- oligose
- lysine
- arginine
- pentapeptide
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K9/00—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof
- C07K9/001—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof the peptide sequence having less than 12 amino acids and not being part of a ring structure
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
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- 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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- Proteomics, Peptides & Aminoacids (AREA)
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- Peptides Or Proteins (AREA)
Abstract
The present invention relates to a kind of oligose peptides and preparation method thereof.In particular it relates to synanthrin Arginine Lysine peptide and preparation method thereof.The oligose peptide of the present invention possesses good stability, simple for process easily operated, can be preferably applied to clinical treatment.
Description
Technical field
The invention belongs to pharmaceutical technology fields, and in particular to a kind of synanthrin Arginine Lysine peptide and preparation method thereof.This
The oligose peptide of invention possesses good stability, simple for process easily operated, can be preferably applied to clinical treatment.
Background technology
Arginine is a constituent in ornithine Xun Huan, has extremely important physiological function.Supplement arginine,
The activity of arginine in liver (arginase) can be increased, help the ammonia in blood being changed into urea and excreted out,
So arginine is powerful to hyperammonemia, hepatic dysfunction etc..Arginine is also the main component of sperm protein, is had
Promote the quality of sperm, improve the effect of sperm motility energy.Arginine is a kind of biradical amino acid, although for adult not
Essential amino acid, but some situations such as body development it is immature or seriously stress under the conditions of, if lack arginine, machine
Body just cannot maintain positive nitrogen equilibrium and normal physiological function；If patient lacks arginine, blood ammonia can be caused excessively high, occurred even
Stupor；If some enzymes of congenital deficiency urea cycle occurs in baby, arginine is also necessary to it, otherwise cannot be maintained
Normal growth and development.So arginic pharmacological action is very more, for example is reducing blood ammonia using ammonia synthesis urea, change
Kind symptom；Promote cell division；Accelerate wound to restore；Discharge and enhancing immune function of ammonia etc. is promoted to have irreplaceable
Effect.
Lysine is amino acid needed by human, is indispensable nutriment.Protein is to form human body cell
Main ingredient, the protein in food enter after human body by the first breaks down into amino acids of digestion, and then human body utilizes lysine again
Rely and the necessary amino acid of human bodies is waited to recombine new human body protein, as immune antiboidy, digestive ferment, plasma protein, growth swash
Element etc..Lysine is most important one kind in the various amino acid of synthetic protein, lacks it, and other amino acid are just limited
It makes or is not used, so also known as the first essential amino acid of human body.Scientist also found that lysine is control growth in humans
Important substance auxin required composition, nervous centralis and peripheral nervous system to people all play an important role.Human body is not
It can itself synthetic lysine, it is necessary to from food draw lysine and be to aid in other nutriments and fully absorbed and utilized by human body
Key substance, human body only has and could improve the absorption and utilization of food protein supplemented with enough lysine, reaches balanced
Nutrition, enhancing development.
Polysaccharide binding protein mainly has following behavior in the mechanism that human body works：(1) it is immunized as caused by humoral factor
Inhibit, such as transforming growth factor(TGF) (TGF)-b or as surgery and the result of chemotherapy；(2) anti tumor immune response is activated, including tree
The maturation of prominent shape cell corrects th1/th2 imbalances, and the white element -15 between monocyte is promoted to produce；(3) by tumour
The direct effect of cell enhances the antitumous effect of chemotherapy by inducing cell apoptosis and inhibition transfer.Polysaccharide combines
Albumen clinic treat on be mainly used for various cancers, in the patient of stomach cancer or colorectal cancer, combine make proteoglycan and
Adjuvant chemotherapy of patients extends the life cycle of patient, this effect is confirmed in multivariate analysis.For Small Cell Lung Cancer,
Proteoglycan combination chemotherapy extends the paracmasis.In addition, proteoglycan is demonstrated to effectively antagonize various cancers, subtract
The harmful effect of few chemotherapy, improves the quality of living.Therefore, it is necessary to study the oligose peptide with immunoregulation effect.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of synanthrin Arginine Lysine groups with immunoregulation effect
Into oligose peptide, which possesses good stability.
Technical scheme is as follows：
The present invention provides a kind of oligose peptide, which is characterized in that the oligose peptide is reacted by synanthrin with peptide to be formed.
Preferably, the peptide is selected from small peptide, preferably dipeptides, tripeptides, tetrapeptide or pentapeptide, more preferable pentapeptide.
In said program, the pentapeptide is made of Arginine Lysine.
Preferably, arginine and lysine ratio are 3 in the pentapeptide:5.
The present invention also provides a kind of methods for preparing oligose peptide, which is characterized in that the described method comprises the following steps：
(1) preparation of Arginine Lysine peptide：Base is inserted into yeast, culture yeasts is expressed, by of expression
It is reacted after grain recycling with endopeptidase Lys-C, the Arginine Lysine peptide of target is obtained after long peptide cleavage；
(2) preparation of synanthrin Arginine Lysine peptide：Synanthrin, Arginine Lysine peptide, water are subjected to Maillard reaction, obtained
To synanthrin Arginine Lysine peptide prod；
(3) freezed after product is concentrated；
(4) it will obtain crushing in synanthrin Arginine Lysine peptide addition pulverizer after crushing obtained by (3), and obtain widow
Glycopeptide powder.
Preferably, the yeast is Pichia pastoris, gives expression to a string of repetition composition sequences by 100 tetrapeptides, amounts to 500
A amino acid, 1500 bases；The particle of recycling is reacted by 3% concentration and 0.5% endopeptidase Lys-C, will contain 500 ammonia
The long peptide of base acid is cut into the pentapeptide containing only 100 amino acid.
It is highly preferred that synanthrin is 25% in the step (2), Arginine Lysine mixture pentapeptide is 1%, pure water is
74%.
The invention further relates to a kind of pharmaceutical compositions, and described pharmaceutical composition is by the oligose peptide described in said program and one
Kind or a variety of pharmaceutically acceptable carriers, diluent or excipient composition.
Have the invention further relates to the oligose peptide or pharmaceutical composition in preparation prevention and/or treatment with protein kinase
Purposes in the disease or disorder agent of pass, the protein kinase are selected from EGFR receptor tyrosine kinases or HER-2 receptor junket
Histidine kinase, the cancer are selected from stomach cancer, colorectal cancer, lung cancer, breast cancer or epidermis squamous carcinoma, preferably Small Cell Lung Cancer.
Specific embodiment
The present invention is further elaborated with reference to embodiment.These embodiments are only in order at purpose of explanation,
And do not limit the scope of the invention and essence.
The preparation of the Arginine Lysine mixture pentapeptide of embodiment 1,1%
(1) preparation of Arginine Lysine peptide：Rink amide MBHA Peptide systhesis resins (0.58mmol/g) are weighed,
It is added in reaction vessel, after being washed successively with dichloromethane and n,N-Dimethylformamide, adds in the N, N- bis- of 20% piperidines
Methylformamide solution mixing is reacted 10 minutes, is repeated aforesaid operations once, is taken off the Fmoc blocking groups of resinous terminal.With N,
After resin after dinethylformamide washing deprotection, side chain and α amino are added in by the lysine of Fmoc radical protections
Fmoc-Lys (Fmoc)-OH, the tert-butyl alcohol, DIC and 2mL n,N-Dimethylformamide fully dissolve.Fmoc-Lys(Fmoc)-
The molar ratio of OH, the tert-butyl alcohol and DIC and resin is 5:1.By reaction vessel sealing be placed in 34 DEG C of constant-temperature tables shaking 4 it is small when,
Reaction end is monitored using Kaiser color reactions.N,N-Dimethylformamide washing resin is added in after completion of the reaction, is taken off simultaneously
Fall two Fmoc protecting groups of lysine, add polypeptide linker, the tert-butyl alcohol, DIC and 2mL n,N-Dimethylformamide, fill
Divide dissolving.The molar ratio of polypeptide linker, the tert-butyl alcohol and DIC and resin is 10:1.The connection raw material of polypeptide is Fmoc-
The arginine of Arg-OH protections.By the reaction vessel of sealing be placed in 34 DEG C of constant-temperature tables shaking 6 it is small when, it is anti-using Kaiser colors
Reaction end should be monitored.DMF washing resins are added in after completion of the reaction, the Fmoc protecting groups on attachment are taken off, with method by amino
Acid is sequentially connected from c-terminus to aminoterminal, forms polypeptide.Tetrapeptide is gone out into a string of weights by 100 tetrapeptides by Pichia anomala expression
Multiple composition sequence, amounts to 500 amino acid, 1500 bases.By in this 1500 bases insertion Pichia pastoris, culture yeasts into
Row expression, the particle of expression is recycled, and the particle of recycling is at 37 DEG C, concentration and 0.5% endopeptidase Lys-C by 3%
React 2 it is small when, this endopeptidase Lys-C cleavage sites are K-Lys, and the long peptide containing 500 amino acid is cut into containing only 100 ammonia
The pentapeptide of base acid obtains Arginine Lysine mixture pentapeptide (arginine:Lysine=3:5).
The preparation of embodiment 2, oligose peptide
(2) preparation of synanthrin Arginine Lysine peptide：By 25% synanthrin, 1% Arginine Lysine mixture pentapeptide (essence
Propylhomoserin：Lysine=3:5), 74% pure water carry out Maillard reaction, reaction condition be 95 DEG C of water-baths, 3 hours of reaction time,
Eventually form brown lotion；
(3) by brown lotion in a nitrogen environment, minus 20 DEG C freeze；
(4) will obtain carrying out carry out Ultramicro-powder in synanthrin Arginine Lysine peptide addition pulverizer after crushing obtained by (3)
It is broken, finally obtain kind of an oligose peptide powder.
Present oligosaccharides peptide powder daily iron supplement amount is 3 grams, the addition available for various food and health products.
Claims (9)
1. a kind of oligose peptide, which is characterized in that the oligose peptide is reacted by synanthrin with peptide to be formed.
2. oligose peptide as described in claim 1, which is characterized in that the peptide be selected from small peptide, preferably dipeptides, tripeptides, tetrapeptide or
Pentapeptide, more preferable pentapeptide.
3. oligose peptide as claimed in claim 2, which is characterized in that the pentapeptide is made of Arginine Lysine.
4. oligose peptide as claimed in claim 3, which is characterized in that arginine and lysine ratio are 3 in the pentapeptide:5.
A kind of 5. method for preparing the oligose peptide described in claim 4, which is characterized in that the described method comprises the following steps：
(1) preparation of Arginine Lysine peptide：Base is inserted into yeast, culture yeasts is expressed, and the particle of expression is returned
It is reacted after receipts with endopeptidase Lys-C, the Arginine Lysine peptide of target is obtained after long peptide cleavage；
(2) preparation of synanthrin Arginine Lysine peptide：Synanthrin, Arginine Lysine peptide, water are subjected to Maillard reaction, obtain chrysanthemum
Saccharin lysine histidine peptide prod；
(3) freezed after product is concentrated；
(4) the obtained synanthrin Arginine Lysine peptide of (3) step is added in pulverizer and crushed, obtain oligosaccharides Gly-His-Lys
End.
6. method as claimed in claim 5, which is characterized in that the yeast is Pichia pastoris, gives expression to a string by 100 four
The repetition composition sequence of peptide amounts to 500 amino acid, 1500 bases；The particle of recycling by 3% concentration with 0.5% it is interior
Peptase Lys-C reacts, and the long peptide containing 500 amino acid is cut into the pentapeptide containing only 100 amino acid.
7. method as claimed in claim 5, which is characterized in that synanthrin is 25% in the step (2), Arginine Lysine mixes
Conjunction object pentapeptide is 1%, pure water 74%.
8. a kind of pharmaceutical composition, described pharmaceutical composition as the oligose peptide any one of claim 1-4 with it is a kind of or
A variety of pharmaceutically acceptable carriers, diluent or excipient composition.
9. oligose peptide or claim 8 described pharmaceutical composition any one of claim 1-4 prepare prevention and/or
Purposes in the treatment disease related with protein kinase or disorder agent, the protein kinase are selected from EGFR receptor tyrosines
Kinases or HER-2 receptor tyrosine kinases, the cancer is selected from stomach cancer, colorectal cancer, lung cancer, breast cancer or epidermis squamous carcinoma, excellent
Select Small Cell Lung Cancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810090447.0A CN108101963A (en) | 2018-01-30 | 2018-01-30 | A kind of oligose peptide and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810090447.0A CN108101963A (en) | 2018-01-30 | 2018-01-30 | A kind of oligose peptide and preparation method thereof |
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| Publication Number | Publication Date |
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| CN108101963A true CN108101963A (en) | 2018-06-01 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
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| CN201810090447.0A Pending CN108101963A (en) | 2018-01-30 | 2018-01-30 | A kind of oligose peptide and preparation method thereof |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102480992A (en) * | 2009-09-30 | 2012-05-30 | 富士胶片株式会社 | Process for production of composition containing collagen peptide |
| CN104379595A (en) * | 2012-06-25 | 2015-02-25 | Hoya株式会社 | Egfr-binding peptide |
-
2018
- 2018-01-30 CN CN201810090447.0A patent/CN108101963A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102480992A (en) * | 2009-09-30 | 2012-05-30 | 富士胶片株式会社 | Process for production of composition containing collagen peptide |
| CN104379595A (en) * | 2012-06-25 | 2015-02-25 | Hoya株式会社 | Egfr-binding peptide |
Non-Patent Citations (1)
| Title |
|---|
| 吴慧伦等: "菊糖与精氨酸的美拉德反应及其产物的抗氧化性能研究", 《食品工业科技》 * |
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Application publication date: 20180601 |
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