Selective manipulation of steroid hydroxyl groups with boronate esters: efficient access to antigenic C-3 linked steroid–protein conjugates and steroid sulfate standards for drug detection

Abstract

The temporary protection of 17alpha-alkyl-5alpha-androstane-3beta,16beta,17beta triols as boronate esters is an efficient method for their regioselective functionalisation. This has been applied to the synthesis of protein-steroid conjugates 7-10 suitable for the development of immunoassays targeting classes of steroids banned from competition in Australian horse racing and other sports. The synthesis of steroids sulfate conjugates 42 and 44 for use as reference standards is also reported.

Chemical sulfation of small molecules—advances and challenges

“…report the regioselective sulfation of a steroid scaffold, especially at the 3- and 16-position in the presence of multiple hydroxyl groups using a boronate ester-based approach 80. The C-3 and C-16 sulfated steroids are desirable as standards for metabolites of anabolic steroid, which are routinely abused in the highly competitive world of sports.…”

The Metabolism of Anabolic-Androgenic Steroids in The Greyhound

“…The organic layer was dried (MgSO4) and concentrated to give a white solid which was purified by flash column chromatography using 10% ethyl acetate in hexane to give the title COCH3), 1.99 (3H, s, COCH3), 0.931 (3H, s, CH3), 0.926 (3H, s, CH3);13 C NMR (100 MHz, CDCl3) δ 214.1, 170.7, 170.4, 74.0, 72.6, 48.8, 47.8, 41.8, 40.7, 35.3, 35.0, 34.8, 32.2, 31.6, 29.7, 26.7, 26.6, 25.0, 23.3, 21.5, 20.9, 19.8, 14.3; HRMS (+EI) calcd for C23H34O5 (M •+ ) 390.2406, found 390.2408. OD) δ 85.0, 80.7, 72.3, 49.6, 47.5, 43.6, 42.0, 37.7, 37.2, 36.4, 35.8, 34.8, 33.3, 31.1, 28.3, 37.4, 23.9, 21.1, 18.2, 15.0; HRMS (+EI) calcd for C20H34O3 .2.7 19-Nor-5β,17α-pregnane-3α,16α,17β-triol and 19-nor-5β,17β-pregnane-3α,16α,17α-triolThe title compound was synthesized on a small scale from 19-noretiocholanolone by a route analogous to that described above (section 2.2.6), with the exception that the 17β-ethyl substituent was introduced via the 17-ketone through a process of ethynylation with lithium acetylide ethylenediamine complex followed by hydrogenation[17]. GC-MS analysis revealed the existence of two peaks in 10:1 ratio which were assigned as the 3α,16α,17β-and 3α,16α,17α-triols respectively based on the susceptibility of the latter to 16,17-isopropylidene acetal formation[18].…”

A Sulfuryl Group Transfer Strategy to Selectively Prepare Sulfated Steroids and Isotopically Labelled Derivatives

“…Current methods to sulfate steroids fall into two main categories (Chart 1). The use of a protected sulfate group (e.g., isobutyl protected sulfate esters) with subsequent deprotection (Simpson and Widlanski, 2006), or the use of a sulfur trioxide equivalent (e.g., chlorosulfonic acid or pyridinesulfur trioxide complex) (Waller and McLeod, 2014), (Hungerford et al, 2006). Although these methods are effective, they suffer from the additional steps of deprotection and/or purification cascades.…”