Unidentified Steroid Hormone of Cushing's Syndrome and Disease

Abstract

A previously unknown HPLC peak was recently observed in urine samples from patients with Cushing's syndrome and disease. We analysed dansylated derivatives of 17keto steroid glucuronides in urine samples from patients with Cushing's syndrome, Cushing disease and from healthy subjects using high-performance liquid chromatography (HPLC) on reversed-phase Cap Cell PakC8. All urine samples from patients with Cushing's syndrome caused by adrenal adenoma and Cushing's disease showed an unknown large peak at the point between [110HE-G] and [110HA-G] peaks and at a retention time of 25.4 min. The same unknown peak was also observed in urine samples from a patient with asymptomatic cortisol-producing adrenal adenoma and two patients with ectopic ACTH-producing tumor, though the peak height was low for the former and one of the latter but high for the second of the two patients. In contrast, healthy male and female urine only showed a very small peak at the same retention time. Urine samples from a Cushing disease treated with op'DDD and Cushing's syndrome bilaterally adrenalectomized and treating with cortisol showed no such peak. The retention time of this unknown peak is clearly different from that of seven 17keto steroid standard glucuronide conjugates. The structure of this substance may be closely related to [110HE-G] or [110HA-G].

Newly Identified Steroid Hormone in Urine of Patients with Cushing's Syndrome: 3.ALPHA.,11.BETA.-Dihydroxy-4-Androsten-17-one

“…Previously, we reported that an "unknown peak substance" was consistently observed in the urine of patients with Cushing's syndrome originating from either adrenal adenoma and/or pituitary adenoma but not in healthy control subjects [4,5]. In the present study we have shown that the "unknown peak substance" is likely to be 3a,11b-DH-A (1A), a known steroid metabolite [9,10,11].…”

“…1. Synthesis of 3a,11b-dihydroxy-4-androsten-17-one (1A) from hydrocortosone acetate (4). Abbreviation: Ac = acetyl, SGC = silica gel chromatography.…”

“…This peak emerges just before the peak for 11b-hydroxy-androsterone (11b-OH-A) in GC and is relatively specific to Cushing's syndrome [5].…”

“…Previously we have reported that the urine of patients with Cushing's syndrome due to adrenal or pituitary adenoma consistently shows an unknown peak in the 17-KS fraction [4][5][6] using high performance liquid chromatography (HPLC) [7] and capillary gas chromatography (GC). This peak emerges just before the peak for 11b-hydroxy-androsterone (11b-OH-A) in GC and is relatively specific to Cushing's syndrome [5].…”

Daily Excretion Levels of an Unidentified Ketosteroid in the Urine of Patients with Cushing's Syndrome and Healthy Subjects Measured by a New Method.

“…The US peak was not detected, or the levels yielded very small peaks in the urine of the healthy subjects [1,5]. Furthermore, we tried hydrolyzing dansyl-US-G with /3-D-glucuronidase and hydrolyzing the resultant dansyl-US with a strong acid to derive US, IWATA et al…”