High-molecular compound, preparation method and application thereof
A preparation, β-inhibitor technology, applied in the field of polymer compound preparation, can solve the problem of inability to improve tumor immunosuppressive microenvironment, abnormal vascular microenvironment infiltrating tumor extracellular matrix density, lack of enhanced CAR-T treatment of lymphoma Treat drug efficacy and other issues, achieve good tumor accumulation effect, increase proliferation and activity, and improve matrix density
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[0068] One, the preparation method of interpolymer, comprises the steps:
[0069] Activate the carboxyl group of polycaprolactone,
[0070] Carboxyl-activated polycaprolactone and hydroxyethyl starch solution were esterified,
[0071] Purified hydroxyethyl starch-polycaprolactone copolymer.
[0072] Specifically, the carboxyl step of activating polycaprolactone is: dissolve polycaprolactone in anhydrous dimethyl sulfoxide, add 1-hydroxybenzotriazole and N-N'-dicyclohexylcarbodiimide, and carry out carboxyl Activation reactions; and / or
[0073] The preparation steps of the hydroxyethyl starch solution are as follows: the hydroxyethyl starch is dissolved in anhydrous dimethyl sulfoxide, preferably, an inert gas protection is used during the dissolution process; and / or
[0074] The temperature of the esterification reaction is 40-80°C. Preferably, an inert gas is used for protection during the reaction.
[0075] In the optimized scheme, the inert gas is helium; the temperatur...
Embodiment 1
[0121] The amphiphilic hydroxyethyl starch-polycaprolactone polymer compound is synthesized according to the following steps:
[0122] (1) Dissolving polycaprolactone and activating its carboxyl group: Dissolving 0.5 g of folic acid with 5 ml of anhydrous dimethyl sulfoxide, then adding 1-hydroxybenzotriazole and N-N'-dicyclohexylcarbodiimide, The carboxyl group activation reaction was carried out at a temperature of 60°C for 30 minutes, and after stirring at room temperature for 2-4 hours, a carboxyl-terminated folic acid solution was obtained; among them, polycaprolactone, 1-hydroxybenzotriazole and N-N'- The molar ratio of dicyclohexylcarbodiimide is 1:1:1;
[0123] 2) Dissolving polysaccharides: Under the protection of helium, fully dissolve 0.5 g of hydroxyethyl starch with a molecular weight of 5 kDa in anhydrous dimethyl sulfoxide at 50 ° C to obtain a dimethyl sulfoxide solution of hydroxyethyl starch:
[0124] 3) Esterification reaction: Mix the carboxy-activated pol...
Embodiment 2
[0132] The present invention uses transwell experiments to study the effect of ICG / LY@HES-PCL on CD-19 + The effect on CAR-T proliferation and activity is as follows: mix raji cells and LY / ICG@HES-CH nanoparticles prepared in Example 1 in advance in a 24-well plate, irradiate with near-infrared light for 5 minutes, and then transfer the cells to The upper chamber of the transwell chamber (pore size ≈ 1 μm), and the lower chamber were planted with CAR-T cells. The transwell chamber was cultured at 37°C and 5% CO2 for 3 days, and then the proliferation of CD-19+CAR-T was analyzed by flow cytometry , detection of CD-19 by enzyme-linked immunoassay + Effect of endocrine IL-2 and TNF-γ on CD-19 after CAR-T culture for 3 days + CAR-T activity was analyzed. The above experiments set ICG, ICG@HES-PCL nanomedicine + near-infrared light irradiation group as the control group.
[0133] After 3 days, the number of CAR T in the LY / ICG@HES-PCL group was 2.7 times that of the control grou...
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