Design,Synthesis And Anticancer Activities Evaluation Of Novel 5H-Dibenzo[b,e]azepine-6,11-dione Derivatives & The Discovery Of A New Method For Synthesis Of Aryl Acetone

Objective:Poly（ADP-ribose）polymer-ase（PARP）which has multiple sub-types is a type of nuclear protease that is involved in the repair of single-stranded DNA damage in most eukaryotic cells.Among the most abundant members of the ribozyme PARP family,PARP-1 plays an important role in the cell life cycle and participates in many cellular processes including DNA repair,replication,transcription,maintenance of genomic stability and regulation of cell death.When the activity of PARP-1 is inhibited by using PARP-1 inhibitors,PARP-1 will dissociate from the gap of single-stranded DNA and block the recruitment of BER repair protein to SSB.The repair SSB replication fork will be suspended in the end of cell division S phase,as a result the replication of cell division S-phase will be in a quiescent state.It can be regarded as a theory that the use of PARP-1inhibitors can effectively control the apoptosis of tumor cells to achieve the therapeutic effect on cancer.In this thesis,PPAR-1 inhibitor Rucaparib and Homoeliythrina were used as lead compounds.Combining with the structural characteristics of PJ34,a series of novel 5H-dibenzo[b,e]azepine-6,11-dione derivatives containing 1,3,4-oxadiazole units were designed and synthesized using computer-assisted design in order to obtain novel structural compounds with antitumor activity,which can lay the foundation for the research and development of new anti-cancer drugs PARP-1 inhibitors.Secondly,when designing and synthesizing target compounds,the key intermediate,anthranilic acid,is not easily available.Therefore,phthalimide which is inexpensive and readily available was used as the raw material,and anthranilic acid was obtained by Hofmann degradation.Based on the structural characteristics of the suitable substrate for Huffman degradation reaction,the scope of application of this reaction was boldly expanded to find a new method for the synthesis of 1-aryl-2-propanone compounds.Methods:1)In-depth analysis of the structure characteristics of the lead compound PARP-1 inhibitors Rucaparib,Homoeliythrina&PJ34 and the mode of interaction with the target,a series of novel 5H-dibenzo[b,e]azepine-6,11-dione derivatives containing1,3,4-oxadiazole units were designed using MOE drug design software through open-loop,closed-loop and skeleton replacement.2)Firstly,the target moleculars were divided into 5H-dibenzo[b,e]azepine-6,11-dione and acylhydrazine by means of retrosynthetic analysis method,and then synthesized separately.Finally,the partially condensed bisacylhydrazines dehydrated to the target compounds.3)The in vivo antumor activity of target compounds to human ovarian cancer cells OVCAR-3 were determined by using MTT assay,Rucaparib was selected as positive control drug.4)The synthesized compounds were tested for their PARP-1 inhibitory activity in vitro by Enzymatic assay to verify whether the designed derivatives act on the intended target.5)Western blotting was used to investigate the mechanism of apoptosis induced by the target compounds.6)By summarizing the characteristics of the existing1-aryl-2-propanone synthesis methods and inspired by the mechanism in which benzylmagnesium chloride is reacted with acetonitrile to form phenylisopropylimine and then hydrolyzed to form 1-phenyl-2-propanone,2-methyl-3-phenylacrylic acid as a starting material was first converted into 2-methyl-3-phenylacrylamide and then the isocyanate was generated by Hofmann degradation.The isocyanate is finally heated to undergo hydrolysis and decarboxylation to give phenylisopropylimine,which ultimately yields 1-phenyl-2-propanone.Results:1)Using computer-assisted drug design,a series of novel5H-dibenzo[b,e]azepine-6,11-dione derivatives containing 1,3,4-oxadiazole units were designed by using the PARP-1 inhibitor Rucaparib and Homoeliythrina as lead compounds through ring-opening,closed-loop,and skeleton replacement.2)The5H-dibenzo[b,e]azepine-6,11-dione-4-carboxylic acid derivative intermediates were synthesized by acylating with phthalic anhydride via using anthranilic acid derivatives as the starting materials and the F-C acylation reaction.5H-dibenzo[b,e]azepine-6,11-dione-4-carboxylic acid derivatives were condensed with various acylhydrazines to prepare bisacylhydrazine intermediates.And then the bisacylhydrazine intermediates were treated with mild dehydrating agent Ts Cl to obtain 40 expected target compounds in 2 series.All the compounds have not been reported in previous literatures and their structures were conformed by ¹H-NMR and ¹³C-NMR.3)The anti-tumor activity of the target compounds was tested by MTT assay.It was found that A19 and A20 showed better antitumor activity than the positive drug Rucaparib.4)Enzymatic assay was used to test PARP-1 inhibitory activity of 40 compounds in 2 series.The results showed that the trend of inhibition to PRAP-1 was basically the same as that of antitumor activities.Both A19 and A20 showed higher potency against PARP-1 than other compounds.5)The effect of A19 and A20 on the expression of Bcl-2 protein in OVCAR-3 cells was investigated.Western blot analysis showed that A19 and A20 could down-regulate the expression of Bcl-2.It suggested that A19 and A20 may induce apoptosis through Bcl-2 pathway.6)The experimental results based on the envisage that2-methyl-3-phenylacrylamide was prepared using 2-methyl-3-phenylacrylic acid as a raw material,followed by Hofmann degradation reaction and hydrolysis to give1-phenyl-2-propanone were not satisfactory.So Curtius rearrangement reaction with the same degradation mechanism as Huffman was subsequently used as the main conversion means to develop a novel“one-pot”method to efficiently synthesize 1-phenyl-2-one.And then the application scope and reaction conditions of the reaction substrate were investigated and optimized.Compared with the reported methods,it has the advantages of easy availability of raw materials,simple operation,mild conditions,short reaction time,and high yield.Conclusion:In this paper,based on a detailed introduction of the extensive physiological activity of PARP-1,the antitumor study and structural features of PARP-1 inhibitors were summarized in detail.Then we took Rucaparib and Homoeliythrina as structural template,designed and synthesized a series of novel 5H-dibenzo[b,e]azepine-6,11-dione derivatives containing 1,3,4-oxadiazole units with anticancer activity at last.In the design and synthesis of target compounds,2-methyl-3-aryl acrylic acid was used as a starting material and Curtius rearrangement reaction was used to transform functional groups as the main means of organic synthesis.On this basis,we carried out the design,verification and optimization of the new method for the synthesis of 1-aryl-2-propanone.