Design, Synthesis, Anti-TMV Activity, and Structure-Activity Relationships of -pregnane C Steroids and Their Derivatives.

-pregnane C steroids exhibit high antiviral activity against the tobacco mosaic virus (TMV). However, the structural modification of -pregnane C steroids and the structure-activity relationship (SAR) of the modified compounds remain unevaluated. Hence, the present study investigated how variations in the original skeletons of natural -pregnane C steroids affect their antiviral activity. A series of glaucogenin C and A derivatives were designed and synthesized for the first time, and their anti-TMV activity was evaluated. Bioassay results showed that most of the newly designed derivatives exhibited good to excellent antiviral activity; among these derivatives, , , and with higher antiviral activity than that of ningnanmycin emerged as new antiviral candidates. Reverse transcription-polymerase chain reaction and Western blotting assay revealed reduced levels of TMV coat protein (TMV-CP) gene transcription and TMV-CP protein expression, which confirmed the antiviral activity of these derivatives. These compounds also downregulated the expression of and . Computational simulations indicated that displayed strong van der Waals energy and electrostatic with the TMV coat protein, affording a lower binding energy (Δ = -56.2 kcal/mol) compared with Ribavirin (Δ = -47.6 kcal/mol). The SAR of these compounds was also evaluated, which demonstrated for the first time that substitutions at C-3 and double bonds of C-5/C-6 and C-13/C-18 are crucial for maintaining high anti-TMV activity.