Synthesized polypeptide and application
A technique for synthesizing peptides, -his-, applied to medical preparations containing active ingredients, peptides, desipeptides, etc., can solve the problems of low bioavailability, tiny TIMPs, difficult to extract, etc., and achieve broad application prospects and therapeutic effects Good results
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Embodiment 1
[0050] Example 1 Synthesis, separation, purification and disulfide bond construction of BSC-1, BSC-2 and BSC-3:
[0051] Synthesized by Fmoc / tBu solid-phase peptide synthesis method (SPPS). The synthesized solid phase support is 2-chlororityl resin, each step of the coupling reaction is one, and the Kaiser test is used to detect the reaction process. After synthesizing the desired peptide chain, use Reagent B (TFA 88%, phenol 5%, H 2 O 5%, TIS 2%) cut off the polypeptide from 2-chlororityl resin, and precipitated with cold ether to obtain the crude polypeptide. The disulfide bonds of the crude product were reduced with mercaptoethanol, then dissolved in DMSO to form a dilute solution, and the intramolecular disulfide bonds were formed by agitating with air overnight.
[0052] RT-HPLC purified polypeptide, C18 column (10mm×200mm, 300 Ȧ particle size), linear gradient elution H 2 O / acetonitrile 0.1% TFA. Freeze-dried to obtain 12mg of SN-1, 13mg of SN-2, and 10mg of SN-3. The...
Embodiment 2
[0053] Example 2 Affinity experiment between substrate and receptor CXCR4
[0054] Cultivate SN-1, 2, 3 and CXCR4+ U373-MAGI-CXCR4 CEM cells, add fluorescein-labeled CXCR4 monoclonal antibody 12G5, incubate for a certain period of time, and measure the fluorescence intensity of the cells. It shows that the polypeptide substrate has a strong affinity with the receptor CXCR4. The IC50 of the sequences BSC-1, BSC-2 and BSC-3 are shown in Table (1):
[0055]
Embodiment 3
[0056] Example 3 The substrate inhibits the interaction between SDF-1α and CXCR4
[0057] Culture fluorescently labeled SDF-1α and CXCR4+ U373-MAGI-CXCR4 CEM cells, add the substrate to be tested, incubate for a certain period of time, and measure the fluorescence intensity of the cells. It shows that the strong affinity between the polypeptide substrate and the receptor CXCR4 inhibits the interaction between the natural ligand SDF-1α and CXCR4, and the IC50 of the sequences BSC-1, BSC-2 and BSC-3 are shown in Table (1).
[0058] The results indicated that the peptide substrate may target the interaction of SDF-1α / CXCR4.
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