C-Met targeting polypeptide and application thereof
A targeting and polypeptide sequence technology, applied in the field of biopharmaceuticals, to achieve the effect of inhibiting tumor growth and proliferation in mice without obvious adverse reactions and side effects
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Embodiment 1
[0031] The polypeptides involved in this embodiment are obtained by solid phase synthesis, and then purity detection is performed by high performance liquid phase HPLC.
[0032] Each polypeptide sequence is as follows: ASP-GLN-ILE-ILE-ALA-ASN-ASN (CM 7-1), ALA-GLN-ILE-ILE-ALA-ASN-ASN (CM 7-2), ASP-GLN-ILEL -Ile-Ala-ASN-ALA (CM 7-3), ASP-GLN-ILE-ILA-ALA-ASN (CM 7-4) or GLN-ILE-ILE-ALA-ASN (CM 7-5), purity Both are 95% or more. See the test results figure 1 . Figure A, CM 7-1 purity is 97.41%. Figure B, CM 7-2 purity is 97.84%. Figure C, CM 7-3 purity is 97.02%. Figure D, cm 7-4 purity is 95.31%, Figure E CM 7-5 purity is 95.43% ..
Embodiment 2
[0034] The test involved in targeted C-MET polypeptides combined with biochemistry by biochemistry, this test, in which one targeted C-MET polypeptide is specifically binding to the target protein at a cell level.
[0035] A polypeptide having prevention and / or inhibiting tumor activity, has an amino acid sequence XA-Gln-Ile-Ile-Ala-ASN-XB, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt having an amino acid sequence XA-Gln-Ile-Ile-Ala-ASN-ELN-ILE-ILA-ALA-ASN-ELN-ILE-ILA-ALA-ASN-ELN-ILE-IL-ALA. The xa, Xb is deleted or replaced with any amino acid.
[0036] In this embodiment, a polypeptide proposed and / or inhibited tumor activity is studied in this embodiment.
[0037] The main target of the active polypeptide having prevention and / or treatment of tumors is C-MET, which is designed to be a polypeptide that targets C-MET to connect FITC fluorescent labels, coupling cells and FITC. The binding ratio of the CM 7-1 polypeptide solution refle...
Embodiment 3
[0046] MTT method detected the series of targeted C-MET series polypeptides inhibiting tumor cell proliferation experiments.
[0047] 3.1 Experimental materials
[0048] Breast cancer cells MDA-MB-231, colon cancer cells HCT-116, pancreatic cancer cells PANC-1, ovarian cancer SKOV-3, liver cancer HepG-2, gastric cancer cells MKN-45, lung cancer cells A549, renal cancer cells CAKI-1 , Cerebral tumor cells U87 and melanoma cells ME, DMEM medium, RPMI 1640 medium, McCoy's 5a medium, penicillin, streptomycin, and fetal bovine serum (FBS), MTT.
[0049] 3.2 Experimental Instrument
[0050] CO 2 Cell incubator, single-channel pipette, inverted microscope, liquid nitrogen tank, small centrifuge, ultra-net platform, automatic high pressure steam sterilization pan, oven, electronic balance, cell counter, 96-well plates.
[0051] 3.3 Experimental Method
[0052] The above various tumor cell concentrations are adjusted to 2 × 10 4 Cells / ml, evenly on 96-well plates, 100 μL / well. After th...
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