Preparation And Antitumor Water-solubility Study Of Two Hydrogen Sulfide Donor Nanoparticles

At present,nano drug-loading systems have been widely used in the delivery of poorly soluble drugs in vivo,which can increase the water solubility and stability of these drugs,thus improving their bioavailability to some extent.In this study,two kinds of nano drug-loading materials were prepared,including drug-loaded micelles prepared from polyethylene glycol and Guerbet hexadecanol and drug-loaded nanoparticles prepared with hyaluronic acid as carrier.There two delivery systems were used to increase the water solubility of the poorly soluble drug hydrogen sulfide donor ADT-OH and curcumin to improve the effect of the drug and the combined therapeutic ability.1?Preparation of reduction-sensitive Guerbet hexadecanol micelles and delivery of anticancer drugsEsterification of Guerbet cetyl alcohol and dithiodipropionic acid to prepare Golbert hexadecanol monoester G₁₆SS grafted at one end,and then the micellar material G₁₆MPGS with reduction sensitivity was conducted,followed by the esterification of the other carboxyl group of G₁₆SS and polyethylene glycol monomethyl ether.The G₁₆MPG micellar material was prepared by carboxylating polyethylene glycol monomethyl ether with glycine as an intermediate and then esterifying with Guerbet hexadecanol.The ADT-OH loaded drug-loaded micelles G₁₆MPGS-ADT and G₁₆MPG-ADT were prepared by the solvent volatilization method using ADT-OH as the target drug.The morphology,drug loading,in vitro release,and stability characterization of the two drug-loaded micelles were investigated.The characterization results showed that the particle size of the drug-loaded micelles was less than 200 nm,the PDI around 0.1⁰.2 and the distribution was uniform.The particle size of the drug-loaded micelles remained relatively stable after being diluted and left at room temperature for a long time;the drug loading of the micelles was more than 4.3%;In vitro release results show that the drug could be released rapidly under the GSH environment.The cytotoxicity and blood safety of drug-loaded micelles in vitro showed that the two blank micelles had no obvious toxicity to tumor cells and normal cells;compared with free drugs,drug-loaded micelles showed excellent inhibition of tumor cell growth in vitro.The tumor cell inhibiting effect of G16MPGS-ADT with reducing sensitivity is better than that of common drug-loaded micelle G16MPG-ADT;the results of apoptosis experiments showed that drug-loaded micelles promoted tumor cell apoptosis in a concentration-dependent manner,which was consistent with the MTT results;The results of in vitro hemolysis experiments showed that the red blood cell hemolysis rate of the blank micelles and drug-loaded micelles was less than the upper limit of 5.0%,which was in line with the requirements of intravenous administration.We constructed a H22 tumor-bearing mouse model to investigate the pharmacodynamics of drug-loaded micelles.The experimental results show that the two drug-loaded micelles have better inhibitory effect on tumor growth than the free drug during the same time of administration.The effect of G₁₆MPGS-ADT micelles with reduction sensitivity was significantly greater than that of G₁₆MPG-ADT micelles.The results of H&E section demonstrate that the tumor tissues of the two drug-loaded micelles showed more necrotic areas than the control group,but there was no damage to the liver and heart of the mice treated with two kinds of drug-loaded micelles.Experiments have shown that we have successfully prepared a safe and reliable micelle material with reduction sensitivity by using Guerbet cetyl alcohol and can better encapsulate the hydrogen sulfide donor ADT-OH drug.The results of in vitro and in vivo experiments show that the drug-loaded micelles we prepared micelles had an outstanding effect in inhibiting tumor cell proliferation.2?Preparation of hyaluronic acid grafted ADT-OH and curcumin nanoparticles and their combinationIn this study,water-soluble hyaluronic acid?HA?was used as drug carrier material to prepare hyaluronic acid curcumin graft?HA-Cur?and hyaluronic acid ADT-OH graft?HA-ADT?by chemical reaction.The two grafts self-assembled in water to form drug-loaded nanoparticles.The grafting rate of the drug was calculated by nuclear magnetic resonance spectroscopy?NMR?,and the particle size of the two nanoparticles,the presence and release of the drug were investigated in vitro.The results showed that the drug grafting rates of the two nanomedicines were HA-Cur7.0%and HA-ADT18.0%,and the particle sizes of the two nanoparticles were between 250-260 nm;The results of DSC showed that both drugs existed in an amorphous state in the nanoparticles,which further indicated that the two drugs were successfully grafted onto hyaluronic acid;the in vitro release results showed that the cumulative release of HA-Cur exceeded 40%within 48 hours,and the release of HA-ADT accumulated more than 30%.In vitro cytotoxicity assays and cell proliferation assays?EDU?of two nanodrugs were performed on human hepatocarcinoma cells 7721,breast cancer cells MB-MDA-231 and MCF-7,which overexpress CD44receptor-bearing tumor cells.The results showed that the inhibitory effect of HA-Cur,HA-ADT nanoparticles on the growth of tumor cells was better than that of free drugs,and the combination of drugs was better than that of single administration.The results of EDU assay of tumor cells showed that the proliferation of the drug nanoparticles group was less than that of the free drug group,and that of the combined drug group was less than that of the single drug group,which was consistent with the results of the cytotoxic MTT assay.The in vitro hemolysis test results of the two nanoparticles showed that the hemolysis rate was less than 4%,indicating that the drug can be used for injection administration.The tumor-bearing mice model was established to evaluate the inhibitory effect of nanoparticles on tumor growth in vivo.The results showed that the tumor growth inhibition was more obvious in the combined administration group during the same period of administration.Little change in mouse body weight indicates that the drug is not toxic or less toxic to mice.From the above experimental results,we could conclude that that the antitumor effect of the two insoluble drugs modified by hyaluronic acid was better than that of free drugs in vitro and in vivo,and the effect of the combined administration was better than that of the single administration.