In Vitro And In Vivo Synergistic Antitumor Activity Of Combined Thiazolidinone In Drug-Resistant Lung Cancer Cells

Lung cancer is the number one cause of cancer-related deaths worldwide, more than 80% of bronchogenic malignancy is from non-small cell lung cancer (NSCLC). Drug resistance and the dose-limiting toxicity are the most important causes of unsuccessful lung cancer chemotherapy. Target therapy is one of the hottest topics in the field of cancer research. Tumorigenesis is quite a complex process with many genes involved. Therefore, single target therapy gradually shows disadvantages. Correspondingly, system biology, which calls for multi-level and multi-target therapy, is accepted. We define a multicomponent drug, rather than consisting of a single compound that interacts with a single target, is a concerted pharmacological intervention of several compounds that interact with multiple targets. Combination drugs that impact multiple targets simultaneously are better at controlling complex disease systems such as cancer. The discovery of new multicomponent drugs with minimal toxicity which are effective against drug-resistant cells is an important step for lung cancer therapy. Thiazolidinone derivatives which are important heterocyclic compounds and found to selectively kill drugresistant cancer cells and induce apoptosis, have potential anticancer applications.OBJICTIVES1. To discovery synergistic anticancer thiazolidinone combination in drug-resistant human non-small cell lung cancer cells by high throughput screening in vitro.2. To study the synergistic antitumor activity of combined thiazolidinone in drug-resistant human non-small cell lung cancer cells and xenograft mouse models. 3. To preliminarily investigate the synergistic antitumor mechanism of combined thiazolidinone.METHODS:1. We previously sythesised a library of thiazolidinone derivatives, designed 100 kinds of thiazolidinone combination (M1-M100), each combination composed four thiazolidinone compounds which selected from the previously sythesised thizolidinone library according to the structure-activity relationship, and these compounds are in the same molar ratio. The anticancer activity of the combinations, was investigated by SRB assay against three cell lines (non-small cell lung cancer cell line H460, paclitaxel-resistant variant H460/TaxR and normal human fibroblasts). We identified one combination with excellent anticancer activity and then investigated its synergistic anticancer activity in vitro.2. The synergistic antitumor activity of combined thiazolidinone was studied in Human H460/TaxR xenograft model which had been built in this thesis.3. We preliminarily investigated the synergistic antitumor mechanism of combined thiazolidinone by tubulin polymerization assay and kinases assay.RESULTS1. One combination (M4) was identified by SRB assay, which can selectively kill H460/TaxR and has minimal toxicity to NHFB. It is composed of four thiazolidinone compounds (27#,167#,107# and 254#) in the same molar ratio.2. In vitro, the GI50 value for M4 is 0.04±0.009μM in H460/TaxR. However, the GI50 value for 27#.167#,107# and 254# are 0.32μM,2.63μmol/L,1.09μmol/L and 0.4μM, whcic are 8.1 to 65 fold compared to M4. In human H460/TaxR xenograft mouse models, the tumor inhibitory rate of single compounds are around 28.7% to 31.3% and the tumor inhibitory rate of M4 is 57.8%, that is higher than the four single compounds.3. 27# and 107# inhibited tubulin polymerization; Kinase assay showed that four thizolidinone compounds targeting different kinds of kinases and the affiliate of compounds targeting the same kinase is also significantly different. higher-affinity bindingCONCLUSIONSWe identified a thiazolidinone combination M4 by high throughput screening, which is composed of four thiazolidinone compounds (27#,167#,107# and 254#) in the same molar ratio. M4 has excellent synergistic anticancer activity in vitro and in vivo. Preliminary investigation shows that M4 may impact several different cellular targets and then result in the synergistic antitumor activity. Our study shows that novel multicompoent anticancer drugs can be found by combination screening in vitro.