The Virtual Screening Of XPO1 Covalent Inhibitors And The Study Of Skin Damage Treatment

Exportin-1 (XPO1) is the most important nuclear transport receptors responsible for nuclear export of many of the major tumor suppress proteins and cell growth regulators. A large number of studies have found that targeted XPO1-mediated nucleocytoplasmic transport mechanism is an effective target on treating various diseases such as cancer, inflammation, skin disease, autoimmune disease, wound healing and anti-virus et al. To the best of our knowledge, most of the existing inhibitors targeting XPO1 aim to block the export of NES-bearing cargoes from nucleus to cytoplasm by covalently binding with the nucleophilic sulphydryl group of Cys528 in the hydrophobic cleft in a manner of Michael-type addition. Cys is the main amino acids of covalent interactions between targeted covalent inhibitors and target protein. Targeted covalent inhibitors has made a significant contribution to the health of people in the past few decades.Compared with the traditional drug, Covalent drugs has some unique advantages such as:stronger affinity and effectiveness, higher selectivity, longer interaction time, good pharmacokinetic properties, etc. Based on the above characteristics, covalent bonding drugs was referred to as the "covalent drugs missile".In this study, we use data mining technology and computational biology such as molecular docking and molecular dynamics simulation technology to discovery targeted covalent drug of XPO1 protein. At the same time, we test the effect of screened compounds in the treatment of skin injury by using UVB radiation skin injury animal model and full-thickness skin excision animal model. The specific research results and conclusions are as follows:(1) A combination of data mining technology and computational biology such as molecular docking and molecular dynamics simulation technology. Targeting active amino acids Cys528 in the XPO1 protein’s pockets combined with NES sequence, we gained a total of 9 XPO1 covalent binding molecules from Pubchem small molecule compounds database and DOCKovalent database. Including three natural molecules, namely Benzyl Isothiocyanate (BITC), Phenethyl Isothiocyanate (PEITC) and Caffeic acid phenethylester (CAPE).(2) Established UVB radiation skin injury animal model, we tested the UV protective effect of the virtual screening small molecule inhibitors PEITC and CAPE. The results showed that minimal erythema dose is 400 mJ/cm2, at this time, △ a< 2. The UV protection of CAPE compounds is weaker than positive control drug alpha mangoatins, but CAPE have UV protection compared with the control group,72 h after irradiation, the A a value of CAPE is 8.43, the control group is 13.13. Statistical analysis found that it has a significant difference. So CAPE has certain UV protection. PEITC compound PEITC compound has no significant difference. So it has no UV protection.(3) Established the full thickness excisional wound healing animal model, we tested the wound healing effect of the virtual screening small molecule covalent inhibitors BITC of XPO1. The results showed wound area of BITC group is less than 5 mm2 in the 12 days after made wound, the wound healing rate is 96%, the wound area of HA group is 10 mm2, the wound healing rate is 88%. However, for model group, the wound area is about 20 mm2, the wound healing rate is lesst than 70%. As a result, BITC has the role of promoting wound healing, and has significant difference compared with the model group.