Discovery Of Anti-hematologic Neoplasmas Candidates That Target BTK

Bruton's tyrosine kinase(BTK)is a non-receptor tyrosine kinase,one of TEC kinase family members,which is the star target of anti-B cells malignant tumor,and has been a hot spot of the drug development.Ibrutinib is the only list drug that target BTK,showing certain therapeutic effect in clinical,but the side effects,such as bleeding and drug resistance,have been reported.The study is based on the great market demand of the BTK inhibitors.We modified and synthesized a variety of analogues of Ibrutinib and performed screening successively at the molecular,cellular and animal levels to abtain candidates.Firstly,we examined the BTK inhibitory activity of all 519 compounds at the molecular level,and 84 compounds that show better biological activity(IC50?10 nM)and kinase selectivity were picked up.Secondly,we performed BTK occupy,Ca2+ flu assays and the inhibition effect of proliferation.The data show that 61 compounds can compeletely occupy BTK at concentration of 100 nM;in the experimennt of Ca2+ flu,the activity of 18 compounds(<100 nM)are weaker than Ibrutinib(10 nM);whereas,the inhibitory activity of some compounds by MTS assay is better than Ibrutinib.Considered the data at cellular level,18 compounds have been chosen to enter the experiment of occupying.The activity of SPI-SIMM-371,353 and 375 are almost equal to Ibrutinib.In acute toxicity,SPI-SIMM-371 and 353 are more secure than 375.So we tested the metabolism activity of our compounds and Ibrutinib,and the metabolism test in BALB/c mice displayed the bioavailability of SPI-SIMM-371 was 100%.In the experiment of acute toxicity,SPI-SIMM-371 is less toxic and more secure than SPI-SIMM-375.Above all,our compounds have an advantage over Ibrutinib in some respects.Finally,we evaluated the chronic pharmacodynamics of SPI-SIMM-371 in tumor models of Mino and OCI-ly10.The data show that 371(12.5,25,50 mg/kg)has better anti-tumor effect than Ibrutinib on the model of Mino;the percentage of inhibition is 37.04%,48.08%,67.88%,respectively.As well as,on the model of OCI-ly10,the activity of 371(5,10,50 mg/kg)is 56.66%,58.61%,69.81%.To summarize,we established the evaluation system at molecular,cellular and animal level based on the covalent binding of Ibrutinib with BTK,influcing the BCR signaling pathways.Through the screening of 519 structure-modified compounds derived from Ibrutinib,we discovery a candidate drug SPI-SIMM-371.SPI-SIMM-371 is a preferable compound with prospective to medicine,its better selectivity to BTK kinase activity in molecular level,far better oral absorption property in PK,and the identical efficacy in vivo at least as much compared with Ibrutinib.This project provide detailed thoughts and a candidate for the research on the discovery and development of BTK targeted anti-hematologic neoplasmas drug and laid the foundation of acquiring the proprietary intellectual property rights at anticancer drug to target BTK.