Design,Synthesis And Bioactivity Study Of The Novel Anticancer Drug Based On The MMP2-Targeted Peptide

Background Cancer thearpy failure is mainly attributed to the chemotherapy drug resistance,metastasis,unbearable side effects and other reasons,so the research for new anticancer drugs is very important.In most types of solid tumors,the MMP2 is upregulated and plays a vital role in inhibition of apoptosis,promotion of invasion,migration and angiogenesis in tumors.The novel anticancer drug targeted to MMP2 has the potential to inhibit tumor metastasis and growth,reduce non-specific side effects.Targeting peptide as a drug delivery has good affinity.In this paper,a peptide recognized and hydrolyzed by MMP2 has been screened,and carried paclitaxel by using chemical and physical strategies to form new targeted anticancer drugs.ObjectiveIn this study,we aimed to 1)screen a peptide which can be recognized and hydrolyzed by MMP2;2)evaluate the anticancer activity in vitro and in vivo of peptide combined with paclitaxel by chemical conjugation and physical inclusion;3)explore novel targeted anticancer to treat cancer and metastasis in clinical.Methods1)Screen MMP2-targeted peptide and preparation of targeted drug: Phage display was applied to screen peptide.The peptides were prepared by automatic peptide synthesizer in using Fmoc-solid-phase synthesis;PDC prodrug and self-assembled drug release delivery was formed by chemical and physical method.2)The abblity of targeted release of novel drugs: Analysis of paclitaxel and self-assembled MMP2 sensing peptide mixture for 2,4,12 and 24 h by HPLC-MS.3)MTT assays: The cytotoxicity of free paclitaxel and new drugs for different cancer cells were determined using MTT according to the dose of paclitaxel were 0.1,0.5,1,5 and 10 ug/ml for 24,48 and 72 h in vitro.4)Scratch assays: The activitation of migration inhibition between free paclitaxel and new drugs were evaluated by using the method of cell scratch assays for 24 h(doses of paclitaxel was 5 ug/ml).5)Treatment of mice bearing human tumor xenografts: The tumor-beared animals were treated with paclitaxel or new drugs by intravenous administration of a specified dose(3 mg/kg body weight)every 5 days.Survival time was recorded in days,and weight and tumor size were monitored three times a week during the study period.6)Computer simulation: The docking simulation between PTX-2'-PVGLIG and MMP2 was achieved by Maestro 9.0,and molecular dynamics simulation of SAMTA7/PTX was obtained by MD.Results1)Screen MMP2-targeted peptide and preparation of targeted drug: We screened a MMP2-targeted peptide [PVGLIG] as a carrier,and obtained the PTX-2'-PVGLIG(PDC)and SAMTA7/PTX(self-assemble)as tnovel targeted drugs.2)The abblity of targeted release of novel drugs: The PTX-2'-PVGLIG can be hydrolyzed by MMP2 and released paclitaxel.3)MTT assays and scratch assays: The antitumor and anti-matastasis activity of PTX-2'-PVGLIG and SAMTA7/PTX were superior to paclitaxel in high expression of MMP2 cells(P<0.05)except for the MTT result in U87MG;and the result in none high expression of MMP2 cells were contrary.4)Anticancer activity in vivo: In vivo,PTX-2'-PVGLIG and SAMTA7/PTX can prolong the survival time in mice and inhibit the tumor size in the mice inoculated with MMP2-overexpressed cells more effectively compared with paclitaxel(P<0.05).5)Computer simulation: PTX-2'-PVGLIG has potential to be hydrolysed by MMP2;paclitaxel can be interacting with SAMTA7 to form core-shell structure.ConclusionsIn conclusion,the novel PTX-2'-PVGLIG(PDC)and SAMTA7/PTX(self-assembly)contain MMP2 targeted peptide carrier were prepared in this study.The two new drugs can be hydrolysed specificity in the presence of MMP2,and they also have good antitumor activity,efficacy and reduce toxicity.Meanwhile PTX-2'-PVGLIG and SAMTA7/PTX can inhibit the migration of MMP2-overexpressed cancer cell,and has the potential to treat MMP2-overexpressed tumor(such as breast cancer,etc.)and tumor metastasis.