Synthsis And Bioassay Of The PEG(Lapatinib & Allosteric PKI-587) Conjugate

Lapatinib is a small molecule oral tyrosine kinase inhibitor that targets human epidermal growth factor receptor 2?HER2,ErbB-2?and epidermal growth factor receptor?EGFR,ErbB-1?,Which can be used in the late stage of metastatic breast cancer due to over-expression of HER2,is a drug that has been approved for marketing.PKI-587,a new drug that is a dual inhibitor of PI3K and mTOR,has been approved by the FDA for clinical treatment of breast cancer.Lapatinib and PKI-587 are both used clinically in the treatment of breast cancer,but both of them have the problems of large side effects,short half-life,low bioavailability,poor solubility.There is synergy when combined with other drugs during the treatment of breast cancer effect.The preliminary experiment of the research group confirmed that Lapatinib and PKI-587 were connected to PEG through different amino acid connecting chains respectively,and the resulting conjugate can improve the solubility of the drug and extend the half-life.Lapatinib's polyethylene glycol conjugate and PKI-587's polyethylene glycol conjugate can improve the solubility of the drug.Combining the literature and the research foundation of the research group in the previous period,we propose an experimental method assumption:Lapatinib and PKI-587 are connected to the same grafting site of the same polyethylene glycol polymer through different amino acid connecting chains,which can more effectively improve the solubility of the drug and extend the half-life of the drug.This article mainly uses the small molecule anticancer drug LPT as the raw m aterial drug,and uses the biodegradable amino acid peptide chains LC-GFLG,GFL G as the connecting chain,after the amidation reaction,the Boc protectionreaction and other reactions are performed,then we can got three protuct intermediates Fmo c-E?LC-GFLG-LPT??OH?,Fmoc-E?GFLG-LPT??OH?;And then in a similar way to g ot the other protuct intermediates:H₂N-LC-GFLG-PKI,H₂N-GFLG-PKI;Then we used Fmoc-E?LC-GFLG-LPT??OH?,Fmoc-E?GFLG-LPT??OH?connected with H₂N-L C-GFLG-PKI,H₂N-GFLG-PKI to got the product E?LC-GFLG-PKI??LC-GFLG-LPT?,E?GFLG-PKI??GFLG-LPT?;a series of reactions were carried out between the prod uct and Boc-Gly-OH,and after separation and purification,we were obtained G-E?L C-GFLG-PKI??LC-GFLG-LPT?,G-E?GFLG-PKI??GFLG-LPT?;Then which is treated with PEG-10K in N,N-dimethylformamide,and stirred at room temperature for a we ek with a low speed in the dark,followed by workup,separation and purification t o obtain the target product PEG-10K-G-E?LC-GFLG-PKI??LC-GFLG-LPT?,PEG-10K-G-E?GFLG-PKI??GFLG-LPT?.The structure of important intermediates and target m olecule were confirmed by ¹H-NMR or MS spectra.The first method of synthesizin gthis kind of polyethylene glycol anticancer drug is the world's first operation.In this article,we use the human-derived breast cancer BT-474 subcutaneous transplantation tumor model to study the anti-tumor activity of target products.The experimental results show that different amino acid linking chains,which have different effects on polyethylene glycol polymer conjugates and human breast cancer cells.But the anti-tumor activity of the target product in the animal solid tumor model is weak,and the therapeutic effect is not obvious.There are two conclusions in this study.First,LPT and allosteric PKI-587 can be simultaneously connected to the same grafting site on the polyethylene glycol polymer.This chemical method of connecting two small molecule drugs to the same polymer carrier at the same time provides a new idea for polymer coupled anticancer drugs.Finally,the different amino acid connecting chains have different effects on the anti-tumor activity of the drugs,but the efficacy is not significant.