Russell Marker, an American chemist, first isolated pregnenolone in 1934 from the sarsaparilla plant Smilax aristolochiifolia. Marker separated pregnenolone from cholesterol using a novel technique he invented called acetolysis, which uses acetic anhydride.

Pregnenolone was first created in 1937 by British physician Gordon Thomas Searle and American endocrinologist Edward Charles Dodds from diosgenin, a sapogenin found in plants like yams and fenugreek.

Pregnenolone was discovered in the 1940s to be a crucial precursor in the biosynthesis of several other natural steroids, such as progesterone, cortisol, and testosterone.

In 1934, Schering AG in Germany introduced the first pregnenolone pharmaceutical preparation under the trade name Anapryl. The drug received American approval in 1938 to treat Addison's disease and in 1941 to treat acne. The use of pregnenolone in medicine has ended. It is still offered as a dietary supplement, where it is marketed as a "natural hormone" and various health benefits are asserted.

A steroid hormone called pregnenolone is created by the body from cholesterol. Progesterone, androstenedione, and testosterone are all precursors to this hormone. Age-related hormonal imbalances may be a result of decreased pregnenolone levels. Some people's memory and cognitive function have been shown to improve with pregnenolone supplementation. Additionally, it might enhance wellbeing, mood, and energy levels. As a dietary supplement, pregnenolone is offered as tablets or capsules.

Pregnenolone is a dietary supplement that can be purchased without a prescription. Typically, it is taken as tablets or capsules. When taken as recommended, pregnenolone is generally regarded as safe.