Hepatitis C virus (HCV), a member of the Flaviviridae family affects approximately 3% of the world population, is becoming the leading cause of liver disease in the world. Therefore, the development of novel and effective treatment strategies to treat chronic HCV infection is urgently needed. A potent HCV inhibitor, DBPR110,is a novel thiazole analogue, which selectively reduced the reporter expression of the HCV1b, HCV2a, HCV3a replicon with an EC50 value of 3.9 ± 0.9 pM, 228.8 ± 98.4 pM and 49.4 ± 12.6 pM, respectively. DBPR110 reduced the activity of HCV1b, HCV2a, and HCV3a replicon with a selective index value of > 12,820,512, > 218,531 and > 1,012,145, respectively. DBPR110 failed to inhibit the other RNA viruses (enterovirus71, dengue, Japanese encephalitis, and influenza viruses) and DNA virus (herpes virus 1) at a concentration more than 10 μM. Sequencing analyses of several individual clones derived from the DBPR110-resistant RNAs purified from cells harboring genotype 1b and 2a HCV replicon revealed that amino acid substitutions mainly within the N-terminal region (domain I) of NS5A were associated with decreased inhibitor susceptibility. In addition, DBPR110 displayed synergistic effects with ribavirin, two NS3 protease inhibitors (telaprevir and boceprevir), and one NS5B polymerase inhibitors (sofosbuvir). A series of in vitro and in vivo safety pharmacology studies showed that DBPR110 had no adverse effects on the central nervous system and respiratory functions. DBPR110 was not mutagenic and clastogenic in vitro and was not clastogenic in an in vivo mouse micronucleus assay. The toxicity/toxicokinetic studies also indicated that the value of no observable adverse effect level (NOAEL) was considered to be safe after 14 daily oral dosing in rats and dogs, respectively. In summary, the pre-clinical results all indicated that DBPR110 is a good drug candidate for further development. Recently, an Investigation New Drug (IND) application of DBPR110 was approved by USFDA for further clinical development in the treatment of patients infected by HCV on Dec 17th, 2015.