RSV604 (RSV-604) is a cell-permeable, non-cytotoxic (CC50 >50 µM) benzodiazepine derivative that is reported to target respiratory syncytial virus (RSV) nucleoprotein (N) via affinity interaction (Kd?= 134 µM) and display similar antiviral potency against 40 clinical human RSV isolates (Ave. EC50 = 800 nM), including both subtypes A and B. Although RSV604 displays host-dependent potency against RSV cellular replication (EC50 = 2 µM/HeLa, 1.8 µM/HEp-2, >50 µM/BHK-21 based on cellular viral F protein level 72 hrs post RSV A2 infection) and release (by >1,000,000-fold/HeLa vs. no effect/BHK-21 72 hrs post infection; [RSV604] = EC90), complete loss of infectivity of released RSV from RSV604-treated cells is observed in both HeLa and BHK-21 cultures. RSV604 is also active against bovine RSV, but inactive toward mouse pneumonia virus, human metapneumovirus or parainfluenza virus types 1 &3 (PIV1 &PIV3).