Manufacture And Anti-tumor Effect Of Nano-drug-loading System Based On Redox-sensitive And Dual-targeting Hyaluronic Acid-methotrexate Prodrug

Nowadays,cancer has become the world's largest disease endangering human health.Chemotherapy is an important method in cancer treatment.However,there still exits some disadvantages of chemotherapeutic drugs,such as the low efficiency of drug targeting,inaccurate control of drug release,serious adverse reactions and other shortcomings.In response to the above problems,we constructed a HA-SS-MTX nano-drug-loading system based on dual-targeting drug delivery.Hyaluronic acid(HA)was a biopolymer with good biocompatibility,which could bind to CD44 receptors as a tumor-targeting ligand.Methotrexate(MTX)was folic acid analogue,which not only could target folate receptor,but also had anti-tumor effect.We used redox-sensitive disulfide bond as the connected bridge to synthesize HA-SS-MTX prodrug.At the same time,HA-SS-MTX prodrug was self-assembled into nanoparticles in deionized water by ultrasound-assisted method.The HA-SS-MTX nano-drug-loading system exhibited uniform size(?110 nm),16.6%drug loading,and good stability.In vitro drug release behavior,HA-SS-MTX nano-drug-loading system presented GSH-induced disassembly and MTX rapidly release behavior(up to 50%in 7 h).In vitro cellular uptake and competition inhibition study proved that HA-SS-MTX nano-drug-loading system could exert dual-specific-targeting effect towards HeLa cells via both CD44/folate receptors-mediated endocytosis.Notably,The IC50 of HA-SS-MTX nano-drug-loading system for HeLa cells was 6.0 ?g/mL,obviously lower than MTX(36.4 ?g/mL)and HA-ADH-MTX nano-drug-loading system(HA-ADH-MTX was conjugated between HA and MTX via non-redox-responsive amide linkage,10.5 ?g/mL).HA-SS-MTX nano-drug-loading system exhibited better cytotoxicity in tumor cells.From fluorescence imaging,we found that the fluorescence signal intensity of the tumor site was stronger than that of other sites through the images of in vivo and ex vitro main organs and tumor tissues,which proved that HA-SS-MTX nano-drug-loading system could accumulate in the tumor tissue.The presence of fluorescent signals in tumor tissue after 48 h post-injection implied that it could exert long-circulating effect.In vivo anti-tumor experiments showed that HA-SS-MTX nano-drug-loading system with less side effects exhibited the most obvious inhibitory effect on tumors after 30 days of observation and measurement of tumor volume in nude mice.In conclusion,HA-SS-MTX nano-drug-loading system would have great potential in targeted therapy of cancer.