iNGR-modified targeted self-assembly RNAi NANO drug delivery system for brain glioma and preparation method of system
A drug delivery system, brain glioma technology, applied in the field of brain glioma targeted self-assembled RNAi nano drug delivery system and its preparation, to achieve the effects of improved delivery efficiency, good endogenous properties, and ideal clinical application prospects
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Embodiment 1
[0036] OEI and DTSSP were dissolved in pure water at a molecular ratio of 1:1, shaken at 35°C for 48 hours to generate pOEI, and unreacted monomers were removed by ultrafiltration. pOEI and PEG containing bifunctional groups were dissolved in PBS at pH 8.0 at a molecular ratio of 1:10, stirred at room temperature for 2 hours, and unreacted substances were removed by ultrafiltration to obtain pOEI-PEG-SH; iNGR cyclic peptide and Sulfo- LC-SMPT was dissolved in PBS at pH 7.2 at a molecular ratio of 1:1, stirred at room temperature for 2 hours, and unreacted substances were removed by ultrafiltration to obtain Sulfo-LC-iNGR. pOEI-PEG-SH and Sulfo-LC-iNGR were mixed according to the molecular ratio of 1:5, reacted at room temperature for 2 hours, and the unreacted substances were removed by ultrafiltration to obtain pOEI-PEG-LC-iNGR, whose hydrogen spectrum was as follows figure 1 (a) shown.
Embodiment 2
[0038] Two primer molecules containing specific sequences and T 4 The ligase was added to the same reaction system, heated at 95°C for 2 minutes, and gradually cooled to room temperature within 1 hour to obtain the transcription template molecule. Appropriate amount of template molecules, AUCG four nucleotides, in vitro RNA transcriptase, etc. were added to the same reaction system, 37°C constant temperature for 28h transcription, DNase was added at 37°C constant temperature for 10min to terminate the transcription reaction, and self-assembled RNAi nanoparticles were obtained.
Embodiment 3
[0040] The pOEI-PEG-LC-iNGR prepared in Example 1 was combined with the spongy and loose RNAi nanoparticles prepared in Example 2 at a mass ratio of 10:1 for 30 seconds to obtain compressed pOEI-PEG-LC-iNGR / RNAi Nanoparticles; gel electrophoresis results show that they can form complete encapsulation when the mass ratio is 10:1, and have specific sensitivity to the high concentration of glutathione in tumor cells (such as figure 1 (c) shown).
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