CN104017055B - A kind of about ST2 protein inhibitor polypeptide and application thereof - Google Patents
A kind of about ST2 protein inhibitor polypeptide and application thereof Download PDFInfo
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- CN104017055B CN104017055B CN201410293553.0A CN201410293553A CN104017055B CN 104017055 B CN104017055 B CN 104017055B CN 201410293553 A CN201410293553 A CN 201410293553A CN 104017055 B CN104017055 B CN 104017055B
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- 239000012268 protein inhibitor Substances 0.000 title claims description 16
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- OGWKCGZFUXNPDA-XQKSVPLYSA-N vincristine Chemical compound C([N@]1C[C@@H](C[C@]2(C(=O)OC)C=3C(=CC4=C([C@]56[C@H]([C@@]([C@H](OC(C)=O)[C@]7(CC)C=CCN([C@H]67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)C[C@@](C1)(O)CC)CC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-XQKSVPLYSA-N 0.000 description 1
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Abstract
The present invention relates to drug world, be specifically related to that there is suppression ST2 protein expression, the polypeptide for the treatment of uterus carcinoma.Its sequence be LLQYDCLALNLHGLRR be brand-new sequence, this polypeptide can vitro inhibition uterine cancer cells Hela cell proliferation, migrate, treat uterus carcinoma；In body lotus tumor model experiment, successfully add the survival rate of mice, there is potential new drug development value.
Description
Technical field
The present invention relates to ST2 protein inhibitor polypeptide and application thereof, be specifically related to that there is suppression ST2 albumen table
Reach, the polypeptide for the treatment of uterus carcinoma.
Background technology
Uterus carcinoma cervical cancer is one of modal malignant tumor of gynecological.Treatment half a year with operation for major-minor, with
The medicines such as radiotherapy, chemotherapy, progesterone estrogen antagonist are auxiliary.But operation is difficult to excise completely cancerous tissue
With subclinical focus.Next is exactly chemicotherapy, but both therapeutic modalities equal normal tissue organ has substantially
Side reaction, further raising curative effect the damage reducing normal tissue are the problems being worth research.The most biological
Targeting therapy on tumor is the most promising, targeted therapy be for tumor generating process blocks certain or multiple
Signal path, reaches to treat the purpose of tumor.
Medical circle also not can be determined that what reason causes uterus carcinoma up to now, it is considered that is probably multinomial
Intersecting caused by synergism of factor, its risk factor has: cervical erosion, sexual behaviour are frequently or property
Life is disorderly or ignores the cleaning of sexual behaviour, ignore menstrual hygiene, sex partner redundant prepuce and may be with bleb
Rash two type virus (HSV_2) and human papilloma (HPS) have substantial connection, even connect sexually transmitted disease (STD), Chlamydia infection
Deng inflammation-related.Blood of cancer patients lymphocyte is in the Th2 dominant state of type cell, and swells
Oncocyte itself is also at the dominant state of Th2, owing to Th1 cell is suppressed, and cellular immune function
Can not effectively activate, antineoplastic immune ability declines, thus is unfavorable for that body tissue is based on cellular immunization
Anti tumor immune response, makes tumor cell existence develop.ST2 is stronger Th2 cell induction agent,
A large amount of releases of Th2 cytokines in uterus carcinoma, facilitate Th0 cell to Th2 direction polarization,
And Th2 polarization can produce the proinflammatory cytokine of substantial amounts of such as IL-6, cause and exacerbate whole body inflammation
Disease is reacted.Additionally, preponderate often along with immunologic tolerance at Th2 type cell, this has also contributed to uterus and has gone out
Blood necrosis increases the weight of.Transmembrane ST2 receptor (ST2L) of the uterus tumor deleting mice may result in the life of tumor
Long and the reduction of transfer, and add the pro-inflammatory cytokine of cyclical level, the NK cell of activation and
CD8+T cell.The rising of ST2 protein function is the key factor that uterus carcinoma is raw.Therefore, suppression ST2
Protein expression, suppression uterus carcinoma development, is the novel targets for the treatment of uterus carcinoma.But, not yet have exploitation maturation
The medicine of the treatment uterus carcinoma of ST2 protein inhibitor polypeptide
ST2 protein inhibitor polypeptide in this patent is proved in uterus carcinoma effectively, to be had in other tumors
The prospect of exploitation in model.
Summary of the invention
Goal of the invention
The present invention provides brand-new sequence, this sequence suppression ST2 protein expression, has uterus carcinoma well
Curative effect.
Technical scheme
ST2 protein inhibitor polypeptide, it is characterised in that its sequence is LLQYDCLALNLHGLRR.
A kind of pharmaceutical composition, it is characterised in that it is pharmaceutically acceptable with more than one that it comprises described polypeptide
Excipient, filler, binding agent, lubricant, disintegrating agent or stabilizer.
Described pharmaceutical composition, it is characterised in that described compositions is injection.
Described agent polypeptide, it is characterised in that effective dose is 10mg/kg.
The application in treatment uterus carcinoma medicine of the ST2 protein inhibitor polypeptide.
Beneficial effect
ST2 protein inhibitor polypeptide, this polypeptide has brand-new sequence, this polypeptide can vitro inhibition uterus carcinoma thin
Born of the same parents' Hela cell proliferation, migrates, and treats uterus carcinoma；Mice is successfully added in body lotus tumor model experiment
Survival rate, there is potential new drug development value.
Detailed description of the invention
The present invention relates to polypeptide by gill biochemical (Shanghai) synthesis.
Embodiment 1
The effect that people uterine cancer cells Hela is migrated by ST2 protein inhibitor polypeptide.
10mg/ml Matrigel (BD company, USA) HELA special culture media is diluted with 1:2, coating
On transwell cell film, room temperature air-dries.The HELA cell trypsin of exponential phase will be cultivated
Digestive system digests, and collects, resuspended with blank HELAs special culture media after washing twice with PBS.At microscope
Lower counting, is adjusted to 1 × 10 by cell concentration5Individual/ml.Prepare the test liquid of each group, with blank HELAs
Special culture media is diluted to 100 μ l.Seed cells in transwell cell, every hole 100 μ l, and will be each
Group test liquid adds in cell.24 orifice plates add 0.6ml thin containing the endothelium of 5% hyclone and 1%ECGS
Born of the same parents' culture fluid stimulates cell migration, in 5%CO2, cultivate 24h for 37 DEG C.Discard Kong Zhongpei liquid, use 90% wine
Essence room temperature fixes 30min, and 0.1% crystal violet room temperature dyeing 10min, clear water rinses, and dabs off with cotton swab
The non-migrating cell of layer, basis of microscopic observation also selects four visuals field to take pictures counting.Suppress according to formula computation migration
Rate (migration inhibition, MI):
Wherein NtestFor the cell migration number of test group, NcontrolCell migration number for blank group.
Test is independently repeated 3 times, and the result that test obtains calculates mean ± SD, and carries out adding up t inspection, * P <
0.05 is significant difference, and * * P < 0.01 is pole significant difference.
Table 1 ST2 protein inhibitor Cys-Lys-Arg-Asp-Trp-Thr-Trp-Cys is to people's uterine cancer cells Hela inhibition of metastasis effect
a:*P<0.05,**P<0.01.
Embodiment 2
The effect that people uterine cancer cells Hela is bred by ST2 protein inhibitor polypeptide.
Use MTT colorimetry.By the Hela cell of logarithmic growth, add 96 holes with 1.0 × 105 and cultivate
In plate, cultivating 24h, experimental port, positive drug control hole are separately added into the Experimental agents ST2 of variable concentrations
Protein inhibitor polypeptide 4 and positive control medicine vincristine；Blank group adds the solvent of same volume.Every hole
If five multiple holes, cultivate 48h, respectively in the every hole of 0h, 2h, 8h, 14h, 20h, 24h, 36h, 48h
Add MTT, after effect 4h, add DMSO, hatch 30min, measure at microplate reader 620nm and inhale
Luminosity A value, by formula growth of tumour cell suppression ratio=(1-experimental group light absorption value/matched group light absorption value) ×
100%.The IC50 calculating Experimental agents is 7.61 μMs.When concentration is 7.61 μMs, to Hela
Big proliferation inhibition rate is 68.34%.
Embodiment 3
ST2 protein polypeptide is to people's uterine cancer cells Hela C57BL/6 black Mus Growth of Tumors Transplanted inhibition test
The tumor tissue taking growth animated period is aseptically milled, and is prepared as 1 × 107Individual/ml cell suspension,
It is inoculated in right side of mice armpit subcutaneous with 0.1ml.Mice-transplanted tumor vernier caliper measurement transplanted tumor diameter, treats
Tumor growth is to 100-200mm3Rear animal random packet.Use the method measuring tumor footpath, dynamically observe ST2
The protein polypeptide inhibition to animal subject tumor.The pendulous frequency of diameter of tumor is every 2 days 1 time, every time
Measure and the most also need to weigh Mus weight.Armpit subcutaneous injection polypeptide on the left of experimental group, negative group gives normal simultaneously
Saline, be administered 14d, its cyclophosphamide subcutaneous administration every other day once, every 3 days subcutaneous administrations one of paclitaxel group
Secondary, polypeptide low dosage one day administered twice group is given twice for one day, and other is respectively organized one day and is administered once.Treatment 14d
After, sacrifice, operation strips tumor mass and weighs.The computing formula of gross tumor volume (tumor volume, TV) is:
TV=1/2 × a × b2
Wherein a, b represent length and width respectively.
Result according to measuring calculates relative tumour volume (relative tumor volume, RTV), calculates
Formula is: RTV=Vt/V0.Wherein V0For (d during sub-cage administration0) measure gained gross tumor volume, VtFor often
Gross tumor volume during one-shot measurement.The evaluation index of anti-tumor activity is Relative tumor rate of increase T/C (%),
Computing formula is as follows:
TRTV: treatment group RTV；CRTV: negative control group RTV.
Test is independently repeated 3 times, and the result that test obtains calculates mean ± SD, and carries out adding up t inspection, * P <
0.05 is significant difference, and * * P < 0.01 is pole significant difference.
Table 2 polypeptide inhibitory action to people's uterine cancer cells Hela C57BL/6 black Mus Growth of Tumors Transplanted
SEQUENCE LISTING
<110>Suzhou Pu Luoda bio tech ltd
<120>a kind of about ST2 protein inhibitor polypeptide and application thereof
<130>
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 16
<212> PRT
<213>artificial sequence
<400> 1
Leu Leu Gln Tyr
Asp Cys Leu Ala Leu Asn Leu His Gly Leu Arg Arg
1 5 10 15
Claims (5)
1.ST2 protein inhibitor polypeptide, it is characterised in that its sequence is LLQYDCLALNLHGLRR.
2. a pharmaceutical composition, it is characterised in that it comprises polypeptide as claimed in claim 1 and more than one pharmaceutically acceptable excipient, filler, binding agent, lubricant, disintegrating agent or stabilizer.
3. pharmaceutical composition as claimed in claim 2, it is characterised in that described compositions is injection.
4. ST2 protein inhibitor polypeptide as claimed in claim 1, it is characterised in that effective dose is 10mg/kg.
5. the ST2 protein inhibitor polypeptide as claimed in claim 1 application in preparation treatment uterus carcinoma medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201410293553.0A CN104017055B (en) | 2014-06-27 | 2014-06-27 | A kind of about ST2 protein inhibitor polypeptide and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410293553.0A CN104017055B (en) | 2014-06-27 | 2014-06-27 | A kind of about ST2 protein inhibitor polypeptide and application thereof |
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| Publication Number | Publication Date |
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| CN104017055A CN104017055A (en) | 2014-09-03 |
| CN104017055B true CN104017055B (en) | 2016-09-14 |
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Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112457392B (en) * | 2021-01-27 | 2021-04-27 | 天津奇云诺德生物医学有限公司 | Soluble ST2 protein antigenic determinant polypeptide and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1331111A (en) * | 2000-06-30 | 2002-01-16 | 上海博德基因开发有限公司 | Polypeptide-Tl/ST2 receptor bindin 10.23 and polynucleotide for coding it |
| WO2013173761A2 (en) * | 2012-05-18 | 2013-11-21 | Amgen Inc. | St2 antigen binding proteins |
-
2014
- 2014-06-27 CN CN201410293553.0A patent/CN104017055B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1331111A (en) * | 2000-06-30 | 2002-01-16 | 上海博德基因开发有限公司 | Polypeptide-Tl/ST2 receptor bindin 10.23 and polynucleotide for coding it |
| WO2013173761A2 (en) * | 2012-05-18 | 2013-11-21 | Amgen Inc. | St2 antigen binding proteins |
Non-Patent Citations (3)
| Title |
|---|
| J Gillibert-Duplantier et al..Gene expression profi * |
| ling identifies sST2 as an effector of ErbB2-driven breast carcinoma cell motility, associated with metastasis.《oncogene》.2011,第31卷3516-3524. * |
| Serum soluble ST2 is associated with ER-positive breast cancer;Da-peng Lu et al.;《BMC Cancer》;20140318;第14卷;摘要部分，第7页左栏第3段 * |
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