Design,Synthesis And Anti-lung Injury Activity Evaluation Of Succinyl Hydroxamate-based As Matrix Metalloproteinase Inhibitors

Matrix metalloproteinases(MMPs)are a family of endogenous proteolytic enzymes,with the dependence of the zinc ion in the structure.Zinc ion is also the active center of these enzymes in playing the role of hydrolysis.The degradation and repair of extracellular matrix(ECM)in normal tissues is co-regulated by MMPs and its natural inhibitors named tissue inhibitors of metalloproteinase(TIMPs).Matrix metalloproteinases begin to overexpress in the stimulation of some specific growth factors and cytokines,leading to its organizational balance disorders with TIMPs.As a result,ECM start to excessively degradate,and the stability of tissue cells is broken,causing a series of diseases such as:Cancer,rheumatoid arthritis,acute lung infection,respiratory distress syndrome,emphysema,arteriosclerosis etc.Therefore,the search for appropriate matrix metalloproteinase inhibitors(MMPIs)may be a effective method in the treatment and prevention of these diseases.In this study,we designed and synthesized Succinyl Hydroxamate derivatives as matrix metalloproteinase inhibitors and evaluate their anti-lung injury activities.The hydroxamic acid group in these structures will cause chelating effect with zinc ion in the structure of MMP,so that the enzyme is inactivated.The design strategy of our molecular is based on succinyl hydroxamate as parent structure with R1 group alkyl substituted(Isobutyl or Hydrogen).We can get novel MMPIs with different aromatic amino acid derivatives duing to R2 and R3 substituents structural transformation.R2 group determines the kinds of amino acids and R3 group determines the derived classes which also affects the hydrophilicity and hydrophobicity of the whole molecular.Ilomastat is a potent,broad-spectrum matrix metalloproteinase inhibitor that can fight against smoking-induced lung injury in mice.And LK-TY803 is a potent compound found in the previous work of our group with the role of prevention and treatment of lung injury.So we choose Ilomastat and LK-TY803 as the positive control drug and evaluate the activity of all new compounds for the prevention and treatment of lung injury.Objective:Synthesizing a series of novel succinyl hydroxamate-based matrix metalloproteinase inhibitors like Ilomastat and LK-TY803.After that,we will evaluate the lung injury activity in order to further improve the design for the respect of some more ideal structures to lay the foundation for the development of new lung injury prevention drugs.Methods:Using isohexyl chloride as the raw material.After reaction with 4S-benzyl-2-oxazolidinone,we can obtain the compound 3 called N-isohexylacyl-4S-benzyloxazole-2-one.And then,in the chiral induction,a nucleophilic substitution reaction occurs to give the alkylation product 4 as a single configuration.Next,compound 5 is prepared by hydrolysis of the amide bond named 2R-isobutyl-1,4-acid-4-tert-butyl ester.Using different types of aromatic amino acid derivatives 8 to condense with the compound 5 respectively to give the intermediate 9,which is removed the t-butyl in the condition of trifluoroacetic acid or cerium trichloride.Finally,objective compound was obtained by the mixed anhydride method.As to the products that have no R1 substitution,we choose ethyl 4-chloro-4-oxobutanoate as the starting material.After hydrolysis and condensation with amino acid derivatives,we can directly obtain the object under alkaline conditions without ester saponification.Selecting mices whose lungs are exposed to PFIB as animal models of lung injury.After intraperitoneal administration by single dose,we choose the survival rate of mice as evaluation index and evaluate the lung injury prevention activity for all new compounds.Results:According to the method described above,we sythesize 19 new target compounds.16 of them have isobutyl substituent and 3 have no substituent as to R1 group.The structures are all confirmed by NMR and their purity is determined by HPLC.Give two examples:Compound LK-TY801:white needle crystal,mp.169-172?,purity:96.42%;Compounds LK-TY820:white powder,mp.154-156?,purity:98.13%.Conclusion:Choosing LK-TY803 and Ilomastat as templates,we design and synthesize 19 new compounds.After activity evaluation,we find that the activity of these compounds has failed to go beyond that of LK-TY803.Therefore,looking for chemical entities which are high activity and good water-soluble still needs further research.