Carolinas Research Day 2021

A NOVEL BIPYRIDONE CISPLATIN ANALOG EXHIBITS ENHANCED ANTICANCER ACTIVITY IN PRECLINICAL MODELS OF UPPER GASTROINTESTINAL CANCERS Samhita Bapat 1 , Vikas Sehdev 2 , Noah Kosnik 2 , Hannah Fischer 2 , Patrick D. Ellis Fisher 2 , and Byron Bennett 3 1 Division of Pharmaceutical Sciences, A & M College of Pharmacy and Health Sciences, Long Island University, Brooklyn, NY 2 Department of Biomedical Sciences, Edward Via College of Osteopathic Medicine, Spartanburg, SC 29303 3 Chemistry Department, Idaho State University, Pocatello, ID

Introduction Upper Gastrointestinal Cancers (UGCs) respond poorly to conventional chemotherapy due to variable P53 status and overactive mechanisms that mediate drug resistance. 1 Platinum-based compounds like Cisplatin (CDDP) are frequently used for treatment of UGCs. 2 However, clinical use of CDDP is limited due to development of drug resistance and dose limiting side-effects resulting in nausea, vomiting, neutropenia, thrombocytopenia and renal toxicity. In this study, we investigated the anticancer potency of a novel CDDP derivative (dichloro [4,40-bis(4,4,4- trifluorobutyl)-2,20-bipyridine] platinum) (DCTF-CDDP) and compared it to CDDP in P53 wild type (P53 WT) and mutant (P53 MT) models of UGCs. Goal of the Study For this study, we evaluated the effect of CDDP or its derivative (DCTF-CDDP) on AGS (P53 WT) and FLO-1 (P53 MT) UGC cell lines. After treatment with CDDP or DCTF-CDDP; MTT cell viability, clonogenic cell survival, cell cycle, Annexin V staining, and western blot analyses were carried out to measure cell viability, cell survival, cell cycle progression, cell death, and expression of apoptotic proteins, respectively, in AGS and FLO-1 UGC cell lines. Our in vitro data indicate that DCTF-CDDP is significantly more potent at inhibiting cell viability and cell survival of P53 wild type and mutant UGC cells. The cell cycle, cell death, and pro-apoptotic protein expression analyses further indicate that DCTF-CDDP is much more effective in suppressing cell cycle progression and inducing apoptotic markers in P53 wild type and mutant UGC cells. Methods and Results

Results

Figure. 2: DCTF-CDDP treatment is significantly more effective than CDDP at inducing SubG1 phase and altering cell cycle progression in UGC cells:

Figure 4. DCTF-CDDP is significantly more effective than CDDP at inducing apoptotic markers in P53 wild type and mutant UGC cells:

Figure 1: DCTF-CDDP is significantly more potent than CDDP against P53 wild type and mutant UGC cells: .

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Conclusions Overall, our study suggests that DCTF-CDDP could be an effective CDDP derivative that can be used to achieve better therapeutic outcome at lower doses and toxic side effects. Future studies characterizing the anticancer and toxicological effects in vivo are warranted to further develop this promising analogue. Fig. 3: (A & B) UGC cells were treated with CDDP or DCTF-CDDP for 8 hrs. and protein expression was evaluated. The data indicate that, compared to CDDP, treatment with DCTF-CDDP mediated enhanced induction of apoptotic proteins in UGC cells. Veh, Vehicle; CDDP, Cisplatin; and DCTF- CDDP, Cisplatin Derivative.

Fig 2: (A & B) The cell cycle data indicate that, compared to CDDP, DCTF-CDDP treatment mediates higher induction of SubG1 phase and suppression of G1 and G2-M phases, respectively, in UGC cells. Veh, Vehicle; CDDP, Cisplatin; and DCTF-CDDP, Cisplatin Derivative. Figure 3. DCTF-CDDP treatment is significantly more effective than CDDP at inducing apoptosis in P53 wild type and mutant UGC cells:

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References

1.Hohenberger P, Gretschel S. Gastric cancer. Lancet. 2003;362:305-315. 2.Grunberger B, Raderer M, Schmidinger M, Hejna M. Palliative chemotherapy for recurrent and metastatic esophageal cancer. Anticancer Res 2007;27:2705 – 14.

Fig. 1: UGC cell lines were treated with various concentrations (0.1-20 µM) of CDDP or DCTF-CDDP. (A & B) The cell viability assay data indicate that a 72 hr. treatment with DCTF-CDDP is significantly more potent and efficacious than that of CDDP. (C & D) The cell survival assay data shows that DCTF-CDDP is more effective than CDDP at suppressing the long term survival of UGC cells. Veh, Vehicle; CDDP, Cisplatin; and DCTF-CDDP, Cisplatin Derivative

Fig 2: (A & B) The FITC-Annexin V flow cytometry data shows that, compared to CDDP, 24 hr. treatment with DCTF-CDDP significantly enhances the percentage of cells in intermediate to late stages of apoptosis . Veh, Vehicle; CDDP, Cisplatin; and DCTF-CDDP, Cisplatin Derivative.

This study is supported by and carried out at the Department of Biomedical Sciences, Edward Via College of Osteopathic Medicine, Gibbs Cancer Center and Department of Chemistry, Idaho State University

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