Preparation And In Vitro Activity Of BSA Docetaxel Targeting Nanoparticles For Lung Cancer

The incidence rate of lung cancer is the highest in malignant tumors.DTX is the primary clinical drug for treating lung cancer.It is effective in treating lung cancer,but it has dose-limiting toxicity.It is mainly due to the poor tumor targeting of DTX and the wide distribution of drugs in the whole body,which leads to the increased toxicity to healthy organs.In order to solve this problem,we designed and constructed a new drug delivery system:4-(2-aminoethyl)benzenesulfonamide,a tumor cell-targeted carbonic anhydrase inhibitor,was coupled with bovine serum albumin(BSA)as a carrier,and DTX was encapsulated by emulsion method to prepare DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles.The preparation conditions of the nanoparticles were optimized by a single factor experiment,and the tumor targeting,activity,and toxicity of the nanoparticles were studied.The results are as follows:(1)Preparation of 4-(2-aminoethyl)benzenesulfonamide modified BSA carrier:BSA and 4-(2-aminoethyl)benzenesulfonamide were coupled by 25%glutaraldehyde to prepare 4-(2-aminoethyl)benzenesulfonamide modified BSA carrier.The coupling degree of the carrier was determined by Ninhydrin colorimetry and ultraviolet spectrophotometry with acetic acid sodium acetate as a buffer.The molecular number of BSA coupled 4-(2-aminoethyl)benzenesulfonamide was 15.(2)Preparation and optimization of DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles:DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles were prepared by emulsification method and optimized by a single factor experiment.The optimum conditions of the BSA nanoparticles are obtained:DTX concentration is 6 mg/mL,coupling protein concentration is 0.6 mg/mL,volume ratio of organic phase to water phase is 1:13,homogenization speed is 6000 rpm,homogenization time is 8 min,homogenization pressure is 600 bar,homogenization times are 5 times.Under the optimal conditions of preparation of the BSA nanoparticles,the particle size of BSA nanoparticles was 87.3±10.5 nm,and the potential was-10.7±1.65 mV.(3)Characterization of DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles:after adding 2%mannitol lyophilized protective agent,the drug loading were 1.6%±0.3%.The BSA nanoparticles were spherical.The crystal morphology of BSA nanoparticles was characterized by X-ray diffraction and differential thermal analysis,which indicated that DTX was amorphous.The stability test showed that DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles was stable during 24 h.The particle size of nanoparticles was within 220 nm.No stratification or precipitation was observed,indicating that the nanoemulsion system was stable.In vitro release experiment,the cumulative release of the DTX group at 96 h was 88.9%,and that of DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles was 3%,the release of the BSA nanoparticles was slower than that of the DTX group.(4)Tumor targeting assay of DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles:the high-definition imaging system and flow cytometry were used to analyze the targeting of the nanoparticles.DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles had higher fluorescence absorption than the control group or DTX@BSA nanoparticle group,which indicated that DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles could target A549 lung cancer cells.(5)Activity and toxicity test of DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles:the inhibition rate of human lung cancer A549 cells was determined by the MTT method.The IC50 of BSA nanoparticles modified with DTX@4-(2-aminoethyl)benzenesulfonamide at 72 h(5.792 μm)was lower than that of DTX(6.499 μm).The maximum tolerated dose of DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles was 60 mg/kg,while that of the positive control(DTX)group was 10 mg/kg,indicating that the toxicity of the nanoparticles was less compared with DTX.In summary,the preparation conditions of DTX@4-(2-aminoethyl)benzenesulfonamide modified BSA nanoparticles are mild and can be amplified on an industrial scale.Preliminary activity and toxicity experiments show that the nanoparticles are worth further development and research,which provides new ideas for treating lung cancer in the clinic.