Ester-bond coupled nano diamond doxorubicin drug and preparation method and application thereof
A technology of diamond doxorubicin and nano-diamonds, which is applied in drug combinations, pharmaceutical formulations, anti-tumor drugs, etc., can solve the problems of strong drug resistance and side effects of tumor cells, and achieve low immunogenicity, stable chemical properties, good solubility effect
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Embodiment 1
[0030] (1) Preparation of nanodiamond-polyethylene glycol (ND-PEG) nanoparticles
[0031] Accurately weigh 10 mg of carboxylated nano-diamonds, pass through dimethyl sulfoxide (DMSO), absolute ethanol (C 2 h 5 OH), sterile water (H 2 O) After washing, place in 10mL MES (0.1M, pH5.8) buffer solution, ultrasonically disperse for 30min to form a suspension, then add 2mg of EDC, 2.5mg of NHS, and stir at room temperature for 6h. Centrifuge for 5 minutes, remove the supernatant, and obtain nano-diamond precipitates with activated carboxyl groups;
[0032] Quickly disperse the nano-diamond precipitate with activated carboxyl group into 10mL BBS (0.1M, pH8.4) buffer solution, ultrasonically disperse for 10min to form a suspension, then add 3mg polyethylene glycol (H 2 N-PEG-COOH), continue to stir and react at room temperature for 12 hours. After the reaction is completed, collect the supernatant by high-speed centrifugation at 15,000 rpm for 5 minutes, and continue to wash with d...
Embodiment 2
[0038] (1)~(2) are prepared with embodiment 1
[0039] (3) Accurately weigh 10 mg of vacuum-dried nano-diamond-polyethylene glycol (ND-PEG-COOH), then add 10 mL of sterilized double-distilled water, ultrasonically disperse for 30 minutes to form a suspension, and then drop by drop into the dissolved 2 mg of doxorubicin-glycidol (DOX-GLY) in sterilized double distilled water, then add 2.0 mg of EDC, 2.5 mg of NHS, and then add a few drops of triethylamine to make the system pH about 8, then place in a water bath at 37 °C The pot was protected from light and stirred for 24 hours. After the reaction was completed, the supernatant was collected by high-speed centrifugation at 15,000 rpm for 5 minutes, washed 5 times with sterilized double-distilled water, and then washed with acetone until the supernatant was colorless. Ethylene glycol-glycidyl-doxorubicin (NPGD) nanomedicine, vacuum-dried and stored in the dark. All supernatants were collected and the volume was recorded for det...
Embodiment 3
[0041] Infrared spectroscopic characterization of nanoparticles
[0042] In order to confirm whether an ester bond is formed between nano-diamond-polyethylene glycol (ND-PEG-COOH) and glycidol-doxorubicin (GLY-DOX), respectively take a small amount of nano-particles and potassium bromide in a mass ratio of 1: 100 were mixed and ground into tablets, and their infrared spectra were measured.
[0043] figure 2 Infrared spectra of the prepared nanoparticles. Doxorubicin (DOX) is compared with the absorption peak of glycidyl-doxorubicin (GLY-DOX), by 3422cm -1 Red shifted to 3339cm -1 , and the absorption peak broadened, indicating that glycidol was successfully connected to the surface of doxorubicin. Compared with nanodiamond (ND), nanodiamond-polyethylene glycol (ND-PEG-COOH) appeared 1621cm -1 and 1656cm -1 double peaks, and this is the characteristic absorption peak of the amide bond, indicating that polyethylene glycol is connected to the diamond surface with an amide b...
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