Neuropeptide and synthesis method and application thereof
A synthesis method and neuropeptide technology, applied in the field of neuropeptides and their synthesis, can solve problems such as body load, toxic injury, temporary cure, etc., and achieve the effect of facilitating degradation, changing activity and state, and improving social memory defects
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[0043] Example 1 Synthesis of the polypeptide of the present invention (TAT-PBD)
[0044] The specific sequence of the polypeptide is: tyrosine-glycine-arginine-lysine-lysine-arginine-arginine-glutamine--arginine-arginine-arginine-histamine Acid-serine-threonine-threonine-arginine-valine (YGRKKRRQRRRHSTTRV)
[0045] 1. Main raw materials: Cl resin, amino acid, DIEA (N,N-diisopropylethylamine), DIC (N,N-diisopropylcarbodiimide), HOBT (1-hydroxybenzotriazole) , DMF (dimethylformamide), Piperidine (piperidine), trifluoroacetic acid, ethanedithiol, phenol, anisole, methanol and water.
[0046] 2. Synthetic ideas:
[0047] Resin: Cl-Resin
[0048] Route: solid phase Fmoc (9-fluorenyl methoxycarbonyl)
[0049] Condensation: DIC+HOBT
[0050] To protect: Piperidine
[0051] Instrument: Fully automatic synthesizer
[0052] 3. Specific steps:
[0053] a. Connect the -COOH of the first amino acid (Vvaline) to Cl-Resin with DIEA, and then block the unreacted functional groups on the resin with methan...
Example Embodiment
[0070] Example 2
[0071] Add this product (TAT-PBD) and the control peptide to the culture medium of mouse hippocampal neurons cultured in vitro for 20 days to a final concentration of 50 μg / ml. One hour later, fix the cells with 4% paraformaldehyde solution and proceed. Immunostaining, as attached figure 1 As shown, it can be observed that the number of dendritic spines of TAT-PBD-treated neurons increases significantly.
Example Embodiment
[0072] Example 3
[0073] Acute brain slices of wild-type mice were prepared, and whole-cell electrophysiological recording was performed in the CA3-CA1 loop of the hippocampus, and TAT-PBD polypeptide (50 mg / ml) was added to the intracellular fluid. Result display (as attached figure 2 (Shown), compared with the control group, PBD can significantly increase the amplitude of LTP.
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