Three new pregnane glycosides, tinctorosides A–C (1–3, resp.), together with one known pregnane glycoside, stephanoside B (4), were isolated from the stems of Marsdenia tinctoria R. Br. (Asclepiadaceae). Their structures were elucidated by extensive spectral methods, especially 2D‐NMR experiments (1H,1H‐COSY, HSQC, HMBC, TOCSY, HSQC‐TOCSY, and ROESY), and chemical evidence.