Design,Synthesis,and Evaluation Of Imidazolo Pyrazine Compounds

Bruton Tyrosine kinase（BTK）is a non-receptor kinase and a key component of B-cell receptor（BCR）signal,which affects the development,maturation and function maintenance of B cells,and plays a crucial role in carcinogenic signal in the proliferation and survival of various B cell malignant tumors.The small molecule inhibitors of this kinase have shown good anti-tumor activity.Although Ibrutinib has significant efficacy in various types of lymphoma and leukemia in clinical practice,there are also some clinical limitations,such as off-target toxicity and drug resistance,which limit its clinical application.Therefore,developing BTK small molecule inhibitors with better selectivity and fewer side effects has become a current research hotspotThis study refers to the structure of ibutinib and innovatively uses imidazolo[1,2-a]pyrazine-8-amine,which has been reported to have kinase inhibitory and anti-tumor activity,as the hinge binding region.At the same time,different linkers and substituents are introduced on the end hydrophobic side chains,and 30 aromatic substituted 8-aminoimidazolo[1,2-a]pyrazine target compounds are designed.The structure of the target compound was confirmed by ¹H NMR,¹³C NMR and mass spectrometry（MS）.The BTK kinase inhibitory activity experiment and in vitro cell proliferation inhibition experiment were carried out to explore its biological activity.The Structure–activity relationship of the compound was analyzed by computer Macromolecular docking method.The biological evaluation results suggested that the compound had weak kinase inhibitory activity against BTK.The inhibitory ratio of compounds DYY-13,DYY-14,DYY-16,and DYY-20 on BTK were 5.6%,7%,11.2%,and11.6%at 100 n M,respectively.In addition,the compounds have good anti-tumor activity,and most of them exhibit a considerable degree of tumor cell growth inhibition rate in MTT experiments.The IC₅₀ values of the four compounds with better inhibition rates,DYY-23,DYY-24,DYY-27,and DYY-28,were 1.991μM,1.387μM,3.006μM,1.813μM.respectively.1Significantly superior to the positive drug ibutinib.The results of Macromolecular docking showed that the selected compounds seemed to exhibit good binding ability to BTK.By analyzing the Structure–activity relationship of the compounds,this study found that the position of N in the piperidine ring and the size and electrical properties of the hydrophobic side chain would affect its kinase inhibition rate;The selection of linkers,substituents,and warheads on the hydrophobic side chains has a significant impact on the anti-tumor activity of the compound.Overall,some compounds exhibit good proliferative inhibitory activity and can be used as potential anti-tumor drugs for further research.