NRP-1 ligand polypeptide-polyethylene glycol-phosphatide compound, active targeting liposome drug delivery system mediated thereby and preparation method thereof
A technology of NRP-1 and phospholipid complexes, which can be used in liposome delivery, pharmaceutical formulations, preparations for in vivo tests, etc., and can solve problems such as no expression
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Embodiment 1
[0044] Example 1: RPARPAR-PEG 3350 -Synthesis, purification and characterization of DSPE
[0045] a) Synthesis, purification and characterization of CRPARPAR
[0046] CRPARPAR polypeptides were synthesized using Boc solid-phase peptide synthesis technology. Weigh 0.4167g of arginine-modified resin (degree of substitution: 0.6mmol / g) into a peptide bottle, swell the resin with DMF for 20min, and drain it. Add about twice the resin volume of TFA and stir for 1 min, remove the TFA, then add TFA and operate in the same way once to remove the Boc protecting group. Use HBTU (benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate) in DMF (N'N-dimethylformamide) solution and DIEA (N,N-di Isopropylethylamine) to activate Boc-Ala, add the resin and shake for 20min. After the reaction was completed, the reaction solution was removed, and the resin was washed with DMF. Subsequently, the remaining amino acids were sequentially connected according to the sequence of CRP...
Embodiment 2
[0049] Example 2: RPARPAR-PEG 2000 -DSPE ( 1,2- Synthesis, Purification and Characterization of Stearoyl Phosphatidylethanolamine)
[0050] Dissolve CRPARPAR polypeptide in PBS solution (pH7.0), take Mal-PEG 2000 - DSPE (maleimide-polyethylene glycol (molecular weight 2000)-1,2-stearyl phosphatidylethanolamine complex) was dissolved in DMF, the two were mixed and reacted with magnetic stirring, HPLC (high performance liquid chromatography ) to monitor the reaction, to be Mal-PEG 2000 - After the DSPE reaction is complete, the reaction is stopped, and excess CRPARPAR and DMF are removed by dialysis (molecular weight cut-off 3.5kDa). Freeze-dried to obtain RPARPAR-PEG 2000 - DSPE, HPLC and FTIR to characterize its structure. The HPLC spectrum shows that the retention time of the main peak moves from about 24 min in Figure A to about 14 min in Figure B; the FTIR spectrum shows that, compared with Figure A, the characteristic peaks of N-H and C=O in Figure B (located a...
Embodiment 3
[0051] Example 3: RPARPAR-PEG 3350 -DOPE( 1,2- Synthesis, Purification and Characterization of Oleoyl Phosphatidylethanolamine
[0052] Dissolve CRPARPAR polypeptide in PBS solution (pH7.0), take Mal-PEG 3350 -DOPE (maleimide-polyethylene glycol (molecular weight 3350)-1,2-oleoylphosphatidylethanolamine complex) was dissolved in DMF, the two were mixed and reacted with magnetic stirring, HPLC (high performance liquid chromatography) Monitor the reaction until Mal-PEG 3350 - Stop the reaction after the DOPE reaction is complete, and remove excess CRPARPAR and DMF by dialysis (molecular weight cut-off 3.5kDa). Freeze-dried to obtain RPARPAR-PEG 3350 -DOPE, HPLC and FTIR characterize its structure. The HPLC spectrum shows that the retention time of the main peak moves from about 24 min in Figure A to about 14 min in Figure B; the FTIR spectrum shows that, compared with Figure A, the characteristic peaks of N-H and C=O in Figure B (located at about 3420 and 1666cm -1...
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