Study On The Preparation Of Curcumin Nanoparticles Based On Hyaluronic Acid Derivatives And Their Antitumor Effect

In recent years,curcumin,extracted from traditional Chinese medicine turmeric,has attracted more and more attention in the field of cancer treatment.As a multifunctional natural phenolic substance,researchers have determined that curcumin has a variety of pharmacological effects,especially an inhibitory effect on breast cancer,stomach cancer,liver cancer,lung cancer and etc.Moreover,curcumin,rich in natural resources and cheap in price,produces few side effects so that it has become a hot issue in the field of medicine.It has also been proven that curcumin has the effect of adjuvant chemotherapy;namely,it can break the defense system of cancer cells and help chemical drugs get inside them.The joint effects of the combination therapy of curcumin and other chemical drugs on prostate cancer,bladder cancer,colon cancer and other tumor cell lines led to a "one plus one is greater than two" effect,far better than the anti-cancer effects of both alone.However,its clinical application is highly limited due to its disadvantages such as instability,poor water solubility,and low bioavailability.The aforementioned problems could be overcome by loading curcumin into nanoparticle carriers with the help of pharmaceutical preparation technology and methods.Hydrogen sulfide?H₂S?,considered as the third gas signaling molecule after carbon monoxide?CO?and nitric oxide?NO?,is closely related to a variety of nervous system diseases.It has been demonstrated that it is involved in the process of functional regulation and plays an important role in the body such as nervous,urinary,endocrine,cardiovascular,respiratory,digestive,reproductive and sensory organs.Its physiological and pathophysiological significance has been the focus of academic research.Recently,people have discovered or synthesized some new H₂ S donor drugs,which also have the biological function of hydrogen sulfide and the efficacy of the drug itself and are able to release hydrogen sulfide slowly in the body environment with a better effect than sodium hydrogen sulfide.In this study,we used water-soluble hyaluronic acid?HA?as the drug carrier material for the preparation of curcumin nanoparticles?HA-Cur?through chemical reactions.Meanwhile,we also prepared a new H₂ S donor prodrug: HA-H₂ S donor conjugate.We then conducted in vitro studies to test the inhibition effects of HA-Cur,HA-H₂ S donor,and the combination of these two conjugates on a few different types of cancer cells.Our results showed that each of these conjugates could inhibit the growth of breast cancer cells with the combination therapy being most active.Objective: To synthesize HA-H₂ S donor and prepare hyaluronic acid derivative based curcumin nanoparticles?HA-Cur?,and to evaluate their in vitro release and in vitro antitumor activity.HA-Cur and HA-H₂ S donor were used to observe the proliferation of H₂ S donor,HA-H₂ S donor and HA-Cur mixture in different cancer cell lines.HA-H₂ S donor,HA-Cur,and the mixture of HA-H₂ S donor and HA-Cur on different cancer cell lines.Research methods:1.HA-ADH was synthesized by the reaction of hyaluronic acid?HA?with adipic acid dihydrazide?ADH?.The HA-ADH was used as the carrier material to react with the Lawson reagent to form HA-H₂ S donor.At the same time,HA-Cur nanoparticles were prepared through the reaction of HA and curcumin.The structures of all the prepared materials were characterized by 1H-NMR and IR.2.The average particle size,polydispersity coefficient?PDI?and zeta potential of the prepared curcumin nanoparticles were investigated by laser particle size and Zeta potential analyzer.The drug loading and the content of flavonoid nanoparticles were determined by high performance liquid chromatography and the drug release behavior in PBS buffer?p H 7.4,containing 0.5% sodium dodecyl sulfate?was examined by dialysis method.Different concentrations of HA-ADH,active pharmaceutical ingredient?curcumin?,HA-H₂ S donor,HA-Cur,and the mixture of HA-Cur & HA-H₂ S donor were investigated by MTT colorimetric method to test their inhibition effect on cancer cell lines.Result: 1.Our results showed that the HA-ADH,HA-H₂ S donor,HA-Cur were successfully synthesized by IR and NMR.The loading degrees of ADH on HA-ADH,H₂ S on HA-H₂ S donor were 61.0%,73.9%,respectively.2.The particle size of HA-Cur was 268.5nm,the loading degree of curcumin on HA was 7.0% and the cumulative in vitro drug release of 96 h was 45.4%.3.The active pharmaceutical ingredient?curcumin?,HA-H₂ S donor,HA-Cur,and the mixture of HA-Cur & HA-H₂ S donor have a certain inhibitory effect on MDA-MB-468 breast cancer cells,and the combination therapy of HA-Cur & HA-H₂ S donor had the best effect.Conclusion: Our results indicated that the HA-Cur and HA-H₂ S donor nanoparticles had uniform particle size distribution,large absolute values of their surface zeta potential and good stability in solution.Our results confirmed that each of these conjugates could inhibit the growth of breast cancers with the combination therapy being most active.