CN104694520A - Hydrolyzed profibrinolysin polypeptide and application thereof - Google Patents
Hydrolyzed profibrinolysin polypeptide and application thereof Download PDFInfo
- Publication number
- CN104694520A CN104694520A CN201510157704.4A CN201510157704A CN104694520A CN 104694520 A CN104694520 A CN 104694520A CN 201510157704 A CN201510157704 A CN 201510157704A CN 104694520 A CN104694520 A CN 104694520A
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- tumor
- hydrolysis
- profibrinolysin
- hydrolyzed
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6424—Serine endopeptidases (3.4.21)
- C12N9/6435—Plasmin (3.4.21.7), i.e. fibrinolysin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/21—Serine endopeptidases (3.4.21)
- C12Y304/21007—Plasmin (3.4.21.7), i.e. fibrinolysin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to the medicine field, in particular to polypeptide capable of promoting profibrinolysin to be hydrolyzed into angiostain and treating a malignant tumor. The sequence of the polypeptide is THELGHLSQKLYGKRSN and is a brand new sequence, the polypeptide can promote the hydrolyzation of the profibrinolysin in vitro and inhibit the proliferation of vascular endothelial cells, in a vivo test, the survival rate of tumor-bearing mice is improved, and the polypeptide has a potential new drug development value.
Description
Technical field
The present invention relates to the former polypeptide of hydrolysis of fibrin lyase and application thereof, be specifically related to that there is promotion proplasmin and be hydrolyzed into angiostatin, the polypeptide for the treatment of malignant tumour.
Background technology
Folkman in 1971 proposes growth and metastasis of tumours and relies on new vessel generation.And after thinking that solid tumor is formed, its development can be divided into without blood vessel phase and two stages of blood vessel phase.Without the blood vessel phase, the existence of minimal neoplastic relies on interstitial disperse blood supply around, and linearly, tumour is limited within 1mm ~ 2mm the speed of growth, and oncocyte number is less than 10
5~ 10
6individual, force tumour to be in " dormant state "; Intravasation after date, tumor tissues is the blood supply of perfusion property, and the exponentially property growth of knurl body, can increase 1.6 ten thousand times after 2 weeks.New vessel not only provides nutrition required for tumour and oxygen, gets rid of meta-bolites, and is the approach of distant metastasis.Therefore, block tumor angiogenesis and just may stop growth and metastasis of tumours, anti-new vessel therapy also one of means becoming oncotherapy.
Angiostatin is the hydrolysate of proplasmin, has stronger angiogenesis inhibiting activity.Large quantity research confirms that angiostatin can suppress growth and the migration of endotheliocyte in vitro, new vessel can be suppressed to be formed in vivo in test, thus inducing apoptosis of tumour cell, Tumor suppression grows, and tumour is in " dormant state ".
Therefore, promote proplasmin hydrolysis, form angiostatin, can effectively suppress vascular endothelial cell proliferation in body, the growth of Tumor suppression.Thus, reach the effect for the treatment of tumour.
At present, do not have the former polypeptide of hydrolysis of fibrin lyase of ripe exploitation to come out, be used for the treatment of malignant tumour.
The former polypeptide of hydrolysis of fibrin lyase in this patent has proved in treatment uterus carcinoma effective, has the prospect developed in other tumor models.
Summary of the invention
Goal of the invention
The invention provides brand-new sequence, this sequence is the former polypeptide of hydrolysis of fibrin lyase, has good curative effect to malignant tumour.
Technical scheme
The former polypeptide of hydrolysis of fibrin lyase, is characterized in that its sequence is THELGHLSQKLYGKRSN.
The former polypeptide of hydrolysis of fibrin lyase treatment malignant tumour, as the application in uterus carcinoma medicine.
Beneficial effect
Utilize the former polypeptide of solid-phase synthesis chemosynthesis hydrolysis of fibrin lyase, this polypeptide has brand-new sequence, and this polypeptide can be hydrolyzed by external promotion proplasmin, and treatment malignant tumour, as uterus carcinoma.The former polypeptide of hydrolysis of fibrin lyase that we find can suppress vascular endothelial cell proliferation vigor simultaneously, and improves tumor-bearing mice survival rate in testing in vivo, has potential new drug development value.
Embodiment
Embodiment 1
The former polypeptide of hydrolysis of fibrin lyase is to the growth of vitro culture human vascular endothelial and survival IC50.
Adopt MTT colorimetry.By the human vascular endothelial HUVEC of logarithmic growth, with 1.0 × 10
5add in 96 well culture plates, cultivate 24h, experimental port, positive drug control hole add the former polypeptide of Experimental agents hydrolysis of fibrin lyase (the raw work synthesis in Shanghai) and the positive control medicine taxol of different concns respectively; Blank group adds the solvent of same volume.Five multiple holes are established in every hole, cultivate 48h, respectively 0h, 2h, 8h, 14h, 20h, 24h, 36h, the every hole of 48h adds MTT, after effect 4h, add DMSO, hatch 30min, absorbance A value is measured, by formula HUVEC growth inhibition ratio=(1-experimental group light absorption value/control group light absorption value) × 100% at microplate reader 620nm place.The IC50 calculating Experimental agents is 7.98 μMs, and taxol IC50 is 5.12 μMs.
Embodiment 2
With vigor in the body of the former polypeptide of tumor model detection hydrolysis of fibrin lyase.
Set up uterus carcinoma tumor model, positive control medicine taxol; Blank group adds the solvent of same volume, and experimental group polypeptide (the raw work synthesis in Shanghai) establishes 3 dosage: 0.75,1.5,3 μMs/Kg.After 21 days, observe mouse survival quantity, calculate survival rate.Result shows, and the former polypeptide of hydrolysis of fibrin lyase can protect small white mouse effectively, and improve the survival rate of tumor-bearing mice, survival rate reaches 78.5%, taxol 31.2.
Embodiment 3
The former polypeptide of hydrolysis of fibrin lyase is to human cervical carcinoma cell HeLa nude mouse xenograft tumor growth inhibition test
The human cervical carcinoma cell HeLa cell strain of taking the logarithm vegetative period, is aseptically prepared into 5 × 10
6/ ml cell suspension, is inoculated in armpit on the right side of nude mice with 0.1ml subcutaneous.With vernier caliper measurement transplanted tumor in nude mice diameter, treat that tumor growth is to 100-200mm
3rear animal random packet.Use the method measuring knurl footpath, dynamically observe the antitumous effect of tested polypeptide.The pendulous frequency of diameter of tumor is every 2 days 1 time, and each measurement also needs weighing mouse heavy simultaneously.Administering mode all adopts tail vein injection.Negative control group injection normal saline, every day 1 time; Cis-platinum group 10mg/kg, Per-Hop behavior 1 time; Administration every day 1 time; Polypeptide is with 20mg/kg administration every day 1 time.Gross tumor volume calculation formula:
TV＝0.52×a×b
2
Wherein a, b represent length and width respectively.Result according to measuring calculates relative tumour volume.The evaluation index of anti-tumor activity is Relative tumor proliferation rate T/C (%), and calculation formula is as follows:
T/C(％)＝T
RTV/C
RTV×100％
T
rTV: treatment group RTV; C
rTV: negative control group RTV
The restraining effect that table 1. polypeptide grows human cervical carcinoma cell HeLa nude mouse xenograft tumor
Polypeptide shows human cervical carcinoma cell HeLa transplanted tumor in nude mice growth inhibition test result, compared with negative control group, the growth of polypeptide group to human cervical carcinoma cell HeLa transplanted tumor has the restraining effect of pole significance compared with positive control cis-platinum, the body weight of polypeptide to laboratory animal has no significant effect, and has no obvious toxic side effects.
SEQUENCE LISTING
Pu Luoda bio tech ltd, <110> Suzhou
<120> former polypeptide of hydrolysis of fibrin lyase and application thereof
<130>
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 17
<212> PRT
<213> artificial sequence
<400> 1
Thr His Glu Leu Gly His Leu Ser Gln Lys Leu Tyr Gly Lys Arg Ser
1 5 10 15
Asn
Claims (3)
1. the former polypeptide of hydrolysis of fibrin lyase, is characterized in that its sequence is THELGHLSQKLYGKRSN.
2. the application of the former polypeptide of hydrolysis of fibrin lyase as claimed in claim 1 in treatment malignant tumor medicine.
3. the application of the former polypeptide of hydrolysis of fibrin lyase as claimed in claim 2 in treatment uterus carcinoma medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510157704.4A CN104694520A (en) | 2015-04-06 | 2015-04-06 | Hydrolyzed profibrinolysin polypeptide and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510157704.4A CN104694520A (en) | 2015-04-06 | 2015-04-06 | Hydrolyzed profibrinolysin polypeptide and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104694520A true CN104694520A (en) | 2015-06-10 |
Family
ID=53342074
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510157704.4A Pending CN104694520A (en) | 2015-04-06 | 2015-04-06 | Hydrolyzed profibrinolysin polypeptide and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104694520A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105039293A (en) * | 2015-08-20 | 2015-11-11 | 倪勤华 | Angiostatin activator polypeptide |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1381460A (en) * | 2002-02-05 | 2002-11-27 | 中国人民解放军空军总医院 | Structure and usage of plasminogen activator (TPA) mutant |
| CN1680430A (en) * | 2005-01-12 | 2005-10-12 | 复旦大学 | Mimic peptide of human plasminogen activator and preparation thereof |
-
2015
- 2015-04-06 CN CN201510157704.4A patent/CN104694520A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1381460A (en) * | 2002-02-05 | 2002-11-27 | 中国人民解放军空军总医院 | Structure and usage of plasminogen activator (TPA) mutant |
| CN1680430A (en) * | 2005-01-12 | 2005-10-12 | 复旦大学 | Mimic peptide of human plasminogen activator and preparation thereof |
Non-Patent Citations (9)
| Title |
|---|
| H. R. LIJNEN等: "Generation of an Angiostatin-like Fragment from Plasminogen by Stromelysin-1 (MMP-3)", 《BIOCHEMISTRY》 * |
| LEVI S. DOWNS JR.等: "Thalidomide and angiostatin inhibit tumor growth in a murine xenograft model of human cervical cancer", 《GYNECOLOGIC ONCOLOGY》 * |
| S. GATELY等: "The mechanism of cancer-mediated conversion of plasminogen to the angiogenesis inhibitor angiostatin", 《PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES》 * |
| YIHAI CAO等: "Kringle 5 of Plasminogen is a Novel Inhibitor of Endothelial Cell Growth", 《THE JOURNAL OF BIOLOGICAL CHEMISTRY》 * |
| 张晓慧等: "血管抑素的研究现状", 《西北国防医学杂志》 * |
| 瞿桂香等: "血红素制备工艺研究进展", 《中央民族大学学报(自然科学版)》 * |
| 蒋立辉: "血管抑素抗肿瘤治疗的研究现状", 《重庆医科大学学报》 * |
| 邵玲莉等: "血管抑素研究进展", 《安徽医药》 * |
| 郑放超等: "血管抑素对小鼠肺腺癌的抑制作用研究", 《中华结核和呼吸杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105039293A (en) * | 2015-08-20 | 2015-11-11 | 倪勤华 | Angiostatin activator polypeptide |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104694520A (en) | Hydrolyzed profibrinolysin polypeptide and application thereof | |
| CN104694521A (en) | Hydrolysis plasminogen polypeptide and application thereof | |
| CN104693281A (en) | Angiostatin activating agent polypeptide and application thereof | |
| CN104726438A (en) | Hydrolysis plasminogen polypeptide and application thereof | |
| CN104004056B (en) | A kind of about Cyclin D protein inhibitor polypeptide and application thereof | |
| CN104045689B (en) | A kind of about somatostatin receptor agonist polypeptide and application thereof | |
| CN104693283A (en) | Angiostatin activating agent polypeptide and application thereof | |
| CN104693282A (en) | Angiostatin activator polypeptide and application thereof | |
| CN104045691B (en) | Npas2 protein agonist polypeptide and application thereof | |
| CN104031128B (en) | A kind of interleukin-33 inhibitor polypeptide and application thereof | |
| CN104017053B (en) | A kind of PER2 protein agonist polypeptide and application thereof | |
| CN104045693B (en) | A kind of about PER2 protein agonist polypeptide and application thereof | |
| CN104045690B (en) | Somatostatin receptor agonist polypeptide and application thereof | |
| CN104693280A (en) | Signal-transducer-and-activator-of-transcription suppressing polypeptide and application thereof | |
| CN104004063B (en) | A kind of about Npas2 protein agonist polypeptide and application thereof | |
| CN104017054B (en) | PER2 protein agonist polypeptide and application thereof | |
| CN104045692B (en) | A kind of Npas2 protein agonist polypeptide and application thereof | |
| CN105039293A (en) | Angiostatin activator polypeptide | |
| CN103417536A (en) | Applications of harmol in preparation of antitumor drugs | |
| CN104017052B (en) | about Npas2 protein agonist polypeptide and application thereof | |
| CN104017055A (en) | Protein ST2 inhibitor polypeptide and application thereof | |
| CN105039294A (en) | Hydrolyzed plasminogen polypeptides | |
| CN104725481A (en) | Vascular endothelial calnexin antagonist polypeptide and application thereof | |
| CN104004060B (en) | Cyclin D protein inhibitor polypeptide and application thereof | |
| CN104059130A (en) | ST2 protein inhibitor related polypeptide and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150610 |
|
| WD01 | Invention patent application deemed withdrawn after publication |