Despite the huge impact of rotavirus infection on global public health, there is no normally available drug against the virus worldwide. We have revealed the interaction of cholesterol metabolism and rotavirus replication, as well as identified statin as a promising drug to repress rotavirus infection, but the medical resources are greatly different across countries, so more drugs are needed for anti-rotavirus treatment in clinical activity. Two cell lines and a human small intestinal organoids were used as the models, which were infected by rotavirus SA11 strain. A clinically derived rotavirus virion, 026K strain, was measured intracellular virus RNA copies in Caco2 cells. We investigated the effects of different cholesterol-lowering drugs, including bisphosphonates (zoledronic acid, ZA), fibrate class (fenofibric acid, FA), vitamin B3 (nicotinic acid, NA), and ezetimibe on rotavirus replication in the pre-clinical models. All these cholesterol-lowering drugs resulted in significant decreases of rotavirus replication. The combinations of FA / ezetimibe with the statins had not the obvious synergies in the inhibition of rotavirus replication than any of them alone. Compared to the other drugs, ezetimibe showed the additional preventive and interference effects towards rotavirus infection. We describe an emerging application of clinical cholesterol-lowering therapy for anti-rotavirus treatment. These results could be directly considered when physicians treat with rotavirus-caused diseases worldwide.