Pancreatic cancer is the fourth highest cause of cancer-related deaths in the United States, with a projected 60,430 new cases diagnosed and 48,220 patients dying in 2021. We employed a small chemical, N-(6-Chloro-2-enzothiazolyl)-3, 4-dimethoxy-benzene propanamide (KY-02111), to target suppression of tumori-sphero-genesis of PANC1ORGRCD19+31+45+133+, to propose a novel therapeutic strategy against drug registrant pancreatic cancer stem cells (PANC1ORGRCD19+31+45+133+). According to our findings, the pancreatic stem cell indicators (CD19+31+45+133+) are found to be more strongly expressed in pancreatic cancer tissues than in normal pancreatic tissues.The flow cytometry, immunoblot and immunofluorescence analysis showed that the expression ofthese markers (CD19+31+45+133+) in PANC1ORGR spheroid cells was lowered by treatment of our new therapeutic approach. Therefore, this study identified the significant relationship of inhibition of tumori-sphero-genesis of PANC1ORGR with associated novel biomarkers (CD19+31+45+133+) which could be target candidates in designing drugs against pancreatic cancer. Further investigation and funding are needed to find the molecular mechanism of inhibition of tumori-sphero-genesis by this small molecule. This work was partly used the financial support from award money of 2017 Translational Research Award, Society of Toxicology and 2018 AACR Minority and Minority-Serving Institution Faculty Scholar award of Dr. Jayanta K. Das.