Application of vesicle nano-drugs in the preparation of brain tumor therapeutic drugs
A nano-drug and therapeutic drug technology, applied in anti-tumor drugs, drug combinations, pharmaceutical formulations, etc., can solve problems such as limiting the therapeutic effect of brain tumors and limited penetration depth of brain tissue, so as to reduce protein adsorption, enrichment and Improved drug release rate, good effect on brain tumor cells and biosafety
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Embodiment 1
[0047] Embodiment 1 Synthetic block copolymer PEG5k-P (DTC2k-TMC15k) and PEG5k-P (DTC2k-TMC15k)-bPEI1.8k
[0048] In a nitrogen glove box, weigh MeO-PEG-OH ( M n = 5.0 kg / mol, 0.50 g, 100 μmol), TMC (1.52 g, 14.55 mmol) and DTC (0.23 g, 1.18 mmol) were dissolved in dichloromethane (DCM, 7.0 mL), and the catalyst diphenyl phosphate was added rapidly with stirring Ester (DPP, DPP / OH molar ratio is 10 / 1). The airtight reactor was sealed and placed in a 40°C oil bath for 2 days under magnetic stirring. Terminate with triethylamine, precipitate twice in glacial ether, filter with suction, and dry in vacuum to obtain PEG5k-P (DTC2k-TMC15k).
[0049] PEG5k-P (DTC2k-TMC15k) is prepared by NPC activation of the terminal hydroxyl chloroformate p-nitrophenyl, and then reacting with the primary amine of branched PEI (bPEI). Specifically, PEG5k-P(DTC2k-TMC15k) (0.4 g, hydroxyl 0.017 mmol) and NPC (50 mg, 0.09 mmol) were dissolved in dry DCM and reacted at 0°C for 24 hours, then precip...
Embodiment 2
[0051] Example 2 Synthesis of targeting polymer
[0052] Targeting polymers can be synthesized in various ways, depending on the terminal functionalization groups of PEG. The synthesis of targeted diblock polymer ApoE-PEG7.5k-P (DTC4.4k-LA19.8k) is divided into two steps, the first step is similar to the synthesis of PEG5k-P (DTC4.4k-LA19.8k), first Replace MeO-PEG-OH with Mal-PEG-OH (Mn = 7.5 kg / mol) ( M n = 5.0 kg / mol) initiated the ring-opening polymerization of DTC and LA to obtain Mal-PEG7.5k-P (DTC4.4k-LA19.8k). In the second step, the ratio of peptide ApoE (sequence Leu Arg Lys Leu Arg Lys Arg Leu Leu Arg Lys Leu Arg Lys Arg Leu Leu Cys) to Mal-PEG7.5k-P (DTC2k-LA15k) is 1.2:1 , ApoE dissolved in DMSO was added dropwise to Mal-PEG7.5k-P (DTC2k-LA15k) dissolved in DMSO under nitrogen, and stirred at 37 degrees for 8 hours. After dialysis with DMSO for 24 hours and then with secondary water for 12 hours, ApoE-PEG7.5k-P (DTC4.4k-LA19.8k) was obtained by freeze-drying....
Embodiment 3
[0057] Example 3 Synthesis of Block Polymer PEG5k-P(TMC15k-DTC2k)-Sp
[0058] PEG5k-P(DTC2k-TMC15k)-NPC synthesized by the same method as in Example 1 was dissolved in 3 mL of DCM, then added dropwise to 3 mL of DCM with spermine (26 mg, 0.13 mmol), and reacted at 30°C for 48 hours Afterwards, dialyze (MWCO 7000) in DCM and methanol (volume ratio 1:1) for 48 hours, precipitate with glacial ether twice, filter with suction, and dry in vacuo to obtain PEG5k-P(DTC2k-TMC15k)-Sp. Yield: 94.7%. NMR and TNBSA showed that the grafting rate of Sp was 97%. Table 1 lists the preparation conditions of each polymer and the NMR characterization results of the product, and the targeting molecule can be connected through the linking group.
[0059] Table 1 The preparation conditions of each polymer and the NMR characterization results of the products
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