Formation of acetamido-Aza-steroids

Beckmann rearrangement of 3α- and 3β-acetamido-5α-androstan-17-one oximes gives homo-aza compounds and the corresponding “second order” Beckmann cleavage ω-cyanoolefin. The mixture of the compounds produced, was separated by column chromatography. The structure of the lactams and the exocyclic nitriles was apparent from ir and nmr spectra.     

Steroidal imidazopyridines and lactam imidazopyridines

Condensation of 17β-acetoxy-2α-bromo-5α-androstan-3-one with unsubstituted and substituted amino-pyridines, gives the corresponding 17β-acetoxy-5α-androstanimidazo[1,2-a]pyridines. Treatment of 16α-bromo-3-aza-A-homo-4α-androsten-4,17-dione with 2-aminopyridine or methyl-2-aminopyridine produces the corresponding 3-aza-A-homo-4α-androsten[16,17:2′,3′]imidazo[1,2-a]pyridines. Similarly, from 2α-bromo-17β-acetamido-5α-androstan-3-one and methylaminopyridine the 17β-acetamido-5α-androstan[2,3:2′,3′]imidazo[1,2-a]methylpyridine has been obtained. The structure of the compounds was apparent from their chemical properties and spectral data (ir, uv and nmr).     

Dissimilar behaviour of 3β, 16α-dihydroxy-5α-androstan-17-one Diacetate under Basic and Acidic Conditions

The base-catalysed rearrangement of 3β, 16α-dihydroxy-5α-androstan-17-one diacetate ( 1 ) in (D₆)benzene/ CD₃OD to 3β, 17β-dihydroxy-5α-androstan-16-one ( 3 ) is followed by ¹³C-NMR spectroscopy. By the same procedure, it is determined that in (D₆)benzene/CD₃OD, but under acid catalysis, 1 does not rearrange to 3 but yields the intermediate product 3β, 16α-dihydroxy-5α -androstan-17-one 17α -methyl hemiacetal ( 5 ).     

Steroidal quinoxalines

The treatment of 3β-hydroxy-16α-bromo-5α-androstan-17-one, 3β-acetoxy-16α-bromo-5-androsten-17-one and 21-bromo-5-pregnen-3β-ol-20-one with 4,5-dimethyl-o-phenylenediamine gave substituted quinoxalines. Hydrolysis of 3β-acetoxy-5-androsteno[16,17-b]-6′,7′-dimethylquinoxaline produced the corresponding 3β-hydroxy compound. 3-Oxo-4-androsteno[16,17-b]-6′,7′-dimethylquinoxaline was obtained by Oppenauer oxidation of the corresponding alcohol.     

Synthese von 4-[3β, 14-Dihydroxy-5β, 14β-androstan-17β-yl]-3-pyrrolin-2-on (Hothesimogenin). Partialsynthetische Versuche in der Reihe der Herzgifte, 11. Mitteilung

Synthesis of 4-[3β, 14-Dihydroxy-5β, 14β-androstan-17β-yl]-3-pyrrolin-2-one (hothesimogenin) We describe the synthesis of 4-[3β, 14-Dihydroxy-5β, 14β-androstan-17β-yl]-3-pyrrolin-2-one (24-aza-24-desoxa-digitoxigenin) (7) , starting from 3-O-acetyl-digitoxigenin (1) .     

5α-androstano [3,2-b]pyridines, 5α-androstano[17,16-b]pyridines and androst-4 and 5-eno[17,16-b]pyridines

A new synthesis of 5α-androstano[3,2-b]pyridin-17β-ol acetate (VIa) and 17-methyl-5α-androstano[3,2-b]pyridin-17β-ol (VIb), first reported by Shimizu, Ohta, Ueno, and Takegoshi, was achieved. The analogous 5α - androstano[17,16-b]pyridin-3β-ol (XII), 5α-androstano[17,16-b]pyridin-3-one (XIVa), and androst-4-eno[17,16-b]pyridin-3-one (XIVb) were also prepared. An illustration of the method follows. Condensation of 3β-hydroxy-5α-androstan-17-one (VIIa) with 3-(2-furyl)acrolein afforded 16-[3-(2-furyl)-2-propenylidene]-3β-hydroxy-5α-androstan-17-one (VIIIa), the oxime (IXa) of which was thermally cyclized to 5α-androstano[17,16-b]-6′-(2-furyl)pyridin-3β-ol (Xa). 3β-Hydroxy-5α-androstano[17,16-b]pyridine-6′-carboxylic acid (XI) was obtained by ozonolysis of Xa. Thermal decarboxylation of XI gave XII. Cinnamaldehyde was used in place of 3-(2-furyl)acrolein to give the corresponding phenylpyridines.     

Combination of carbon isotope ratio with hydrogen isotope ratio determinations in sports drug testing

Carbon isotope ratio (CIR) analysis has been routinely and successfully applied to doping control analysis for many years to uncover the misuse of endogenous steroids such as testosterone. Over the years, several challenges and limitations of this approach became apparent, e.g., the influence of inadequate chromatographic separation on CIR values or the emergence of steroid preparations comprising identical CIRs as endogenous steroids. While the latter has been addressed recently by the implementation of hydrogen isotope ratios (HIR), an improved sample preparation for CIR avoiding co-eluting compounds is presented herein together with newly established reference values of those endogenous steroids being relevant for doping controls. From the fraction of glucuronidated steroids 5β-pregnane-3α,20α-diol, 5α-androst-16-en-3α-ol, 3α-Hydroxy-5β-androstane-11,17-dione, 3α-hydroxy-5α-androstan-17-one (ANDRO), 3α-hydroxy-5β-androstan-17-one (ETIO), 3β-hydroxy-androst-5-en-17-one (DHEA), 5α- and 5β-androstane-3α,17β-diol (5aDIOL and 5bDIOL), 17β-hydroxy-androst-4-en-3-one and 17α-hydroxy-androst-4-en-3-one were included. In addition, sulfate conjugates of ANDRO, ETIO, DHEA, 3β-hydroxy-5α-androstan-17-one plus 17α- and androst-5-ene-3β,17β-diol were considered and analyzed after acidic solvolysis. The results obtained for the reference population encompassing n?=?67 males and females confirmed earlier findings regarding factors influencing endogenous CIR. Variations in sample preparation influenced CIR measurements especially for 5aDIOL and 5bDIOL, the most valuable steroidal analytes for the detection of testosterone misuse. Earlier investigations on the HIR of the same reference population enabled the evaluation of combined measurements of CIR and HIR and its usefulness regarding both steroid metabolism studies and doping control analysis. The combination of both stable isotopes would allow for lower reference limits providing the same statistical power and certainty to distinguish between the endo- or exogenous origin of a urinary steroid.     

Neurosteroids: 7-aza-allopregnanolone—a poor substitute for allopregnanolone

7-Nor-20-oxopregn-5-en-3β-yl acetate was converted into (20R)-5β,6β-epoxy-7-nor-5β-pregnane-3β,20-diyl diacetate in three steps. Stereospecific migration of the 6α-hydride ion led to a 6-oxo derivative with a 5α-configuration. The (Z)-oxime of this ketone underwent Beckmann rearrangement to yield a lactam with the nitrogen in position 7. Lithium aluminium hydride reduction yielded the dihydroxy amine, which was either oxidised or Boc-protected and then oxidised to 7-aza-5α-pregnane-3,20-dione. Its regioselective reduction produced 7-aza-3α-hydroxy-5α-pregnan-20-one—a poor inhibitor for the binding of [³⁵S]TBPS to the GABA_A receptor. The corresponding lactam—7-aza-3α-hydroxy-5α-pregnane-6,20-dione—was inactive.     

Steroids and sex hormones. The synthesis of 1-beta, 4-beta-oxido-3-aza-17-beta-hydroxy-A-homo-5-alpha-androstane

A sterically controlled transformation of 3-oxo-17β-acetoxy-Δ^1?-α-androstene ( 2 ) into 1β,4β-oxido-3-aza-17β-hydroxy-A-homo-5α-androstane ( 16 ) is described. With the exception of two conversions [ 14 → 15 (60%); 15 → 16 (50%)], the yields of the remaining seven steps are higher than 75% each. The reaction sequence will serve as a model for an analogous partial synthesis of samandarine ( 1 ).     

Thiosteroids—XXXV : Diastereomers of steroidal thiolsulfinates

17β-Hydroxy-5α-androstan-2α,3α-anti(R)-episulfoxide on treatment with methanol and ethanol in the presence of a trace amount of sulfuric acid gave diastereomers of bis((2β-methoxy- and 2β-ethoxy-17β-hydroxy-5α-androstan-3α-yl) disulfide S-monoxides respectively. The absolute configuration of the compounds was established by their Grignard reactions leading to diastereomeric phenyl sulfoxides stereoaspecifically.     

Effect of changes in the deuterium content of drinking water on the hydrogen isotope ratio of urinary steroids in the context of sports drug testing

The hydrogen isotope ratio (HIR) of body water and, therefore, of all endogenously synthesized compounds in humans, is mainly affected by the HIR of ingested drinking water. As a consequence, the entire organism and all of its synthesized substrates will reflect alterations in the isotope ratio of drinking water, which depends on the duration of exposure. To investigate the effect of this change on endogenous urinary steroids relevant to doping-control analysis the hydrogen isotope composition of potable water was suddenly enriched from -50 to 200 ‰ and maintained at this level for two weeks for two individuals. The steroids under investigation were 5β-pregnane-3α,20α-diol, 5α-androst-16-en-3α-ol, 3α-hydroxy-5α-androstan-17-one (ANDRO), 3α-hydroxy-5β-androstan-17-one (ETIO), 5α-androstane-3α,17β-diol, and 5β-androstane-3α,17β-diol (excreted as glucuronides) and ETIO, ANDRO and 3β-hydroxyandrost-5-en-17-one (excreted as sulfates). The HIR of body water was estimated by determination of the HIR of total native urine, to trace the induced changes. The hydrogen in steroids is partly derived from the total amount of body water and cholesterol-enrichment could be calculated by use of these data. Although the sum of changes in the isotopic composition of body water was 150 ‰, shifts of approximately 30 ‰ were observed for urinary steroids. Parallel enrichment in their HIR was observed for most of the steroids, and none of the differences between the HIR of individual steroids was elevated beyond recently established thresholds. This finding is important to sports drug testing because it supports the intended use of this novel and complementary methodology even in cases where athletes have drunk water of different HIR, a plausible and, presumably, inevitable scenario while traveling.     

Photochemical Rearrangement of a Steroidal α, β-Epoxylactam. Photochemical reactions,XII [1]

The UV. irradiation of 17β-hydroxy-4α, 5α-epoxy-2-azaandrostan-3-one ( 1 ) yields 17β-hydroxy-2-aza-10 (5 → 4-abeo)-4ζ (H)-androsta-3,5-dione ( 3 ).     

Reaction of a 5α-bromo-6β,19-epoxysteroid with BF3·Et2O/Ac2O: An evidence of a cyclic bromonium cation

Treatment of 3β-acetoxy-5-bromo-6β,19-epoxy-5α-androstan-17-one with Ac₂O and BF₃·OEt₂, produced the cleavage of the epoxy moiety and migration of the bromine atom to afford 3β,19-diacetoxy-6α-bromo-5-hydroxy-5β-androst-17-one in high yield.     

Metabolic studies of mesterolone in horses

Mesterolone (1α-methyl-5α-androstan-17β-ol-3-one) is a synthetic anabolic androgenic steroid (AAS) with reported abuses in human sports. As for other AAS, mesterolone is also a potential doping agent in equine sports. Metabolic studies on mesterolone have been reported for humans, whereas little is known about its metabolic fate in horses. This paper describes the studies of both the in vitro and in vivo metabolism of mesterolone in racehorses with an objective to identify the most appropriate target metabolites for detecting mesterolone administration.In vitro biotransformation studies of mesterolone were performed by incubating the steroid with horse liver microsomes. Metabolites in the incubation mixture were isolated by liquid-liquid extraction and analysed by gas chromatography-mass spectrometry (GC-MS) after acylation or silylation. Five metabolites (M1-M5) were detected. They were 1α-methyl-5α-androstan-3α-ol-17-one (M1), 1α-methyl-5α-androstan-3β-ol-17-one (M2), 1α-methyl-5α-androstane-3α,17β-diol (M3), 1α-methyl-5α-androstane-3β,17β-diol (M4), and 1α-methyl-5α-androstane-3,17-dione (M5). Of these in vitro metabolites, M1, M3, M4 and M5 were confirmed using authentic reference standards. M2 was tentatively identified by mass spectral comparison to M1.For the in vivo metabolic studies, Proviron^® (20 tablets × 25 mg of mesterolone) was administered orally to two thoroughbred geldings. Pre- and post-administration urine samples were collected for analysis. Free and conjugated metabolites were isolated using solid-phase extraction and analysed by GC-MS as described for the in vitro studies. The results revealed that mesterolone was extensively metabolised and the parent drug was not detected in urine. Three metabolites detected in the in vitro studies, namely M1, M2 and M4, were also detected in post-administration urine samples. In addition, two stereoisomers each of 1α-methyl-5α-androstane-3,17α-diol (M6 and M7) and 1α-methyl-5α-androstane-3,16-diol-17-one (M8 and M9), and an 18-hydroxylated metabolite 1α-methyl-5α-androstane-3,18-diol-17-one (M10) were also detected. The metabolic pathway for mesterolone is postulated. These studies have shown that metabolites M8, M9 and M10 could be used as potential screening targets for controlling the misuse of mesterolone in horses.     

The anaerobic degradation of deoxycholic acid by PSEUDOMONAS SP. NCIB 10590

The anaerobic metabolism of deoxycholic acid by Pseudomonas sp. NCIB 10590 was studied. The metabolic pathway was similar to that operating under aerobic conditions with 12β-hydroxyandrosta-1,4-dien-3,17-dione as the major neutral product an metabolites which are not produced during aerobic metabolism were isolated and evidence is presented for the following structures: 9α-hydroxyandrost-1-en-3,17-dione, 12α,17)β-dihydroxyandrosta-1,4-dien-3-one; 3β,12β-dihydroxy-5β-androstan-17-one an formation and significance of the phenolic secosteroid is discussed.     

Synthesis,x-ray structure and molecular mechanics studies of the boar taint steroid (5α -androst-16-en-3-one)

5α-Androst-16-en-3-one has been prepared from 5α-androstan-3β-ol-17-one in an overall yield of 34% by the vinyl iodide route. Accurate molecular dimensions have been determined by X-ray crystal structure analysis and by molecular mechanics calculations. There is significant twisting of the angular methyl groups in the molecule.     

Synthesis of Atamestane (SH 489): An Aromatase Inhibitor

Atamestane (SH 489) was synthesized from 17β-Acetoxy-1α-methyl-5α-androstan-3-one. Thus dibromination followed by dehydrobromination of the A-ring gave dienone (5) in good yield, which was hydrolyzed and oxidized to give the title compound in 60% overall yield.     

Synthesis and biological evaluation of 3-beta,20(R)-dihydroxy-protost-24-ene

The possible lanosterol precursor, 3β, 20(R)-dihydroxy-protost-24-ene ( 1 a ) has been prepared, in thirteen steps, from 3α-hydroxy-4α, 8, 14-trimethyl-18-nor-5α, 8α, 9β, 13ξ, 14β-androstan-17-one (mixture of 2 and 3 ). In vitro experiments with rat liver homogenates failed to convert 1 a to lanosterol.     

Steroids and steroidases—XIX : Comparison of diazomethane and tiffeneau-demjanov homologations of 5α-3-oxosteroids. Evidence for predominant equatorial approach of the C-3 carbonyl group by diazomethane

Quantitative comparisons of the product ratios of the mechanistically similar diazomethane and Tiffeneau-Demjanov homologations of 17β-hydroxy-5α-androstan-3-one and 5α-cholestan-3-one have shown that equatorial approach of diazomethane to the C-3 CO group predominates to the extent of 70–79%. The data for both the C-17β-OH and -C₈H₁₇ substituted steroids are in close agreement thereby confirming that the C-17 substituents do not exert any significant long range effect on the reactions studied.     

On the formation of homo-azasteroidal esters of N,N-bis(2-chloroethyl)aminobenzoic acid isomers and their antitumor activity

The N, N-bis(2-chloroethyl)aminobenzoate isomers and the 4-methyl-3-N, N-bis(2-chloro-ethyl)aminobenzoate of 3β-hydroxy-13α-amino-13,17-seco-5α-androstan-17-oic-13,17-lactam, 3α-hydroxy-13α-amino-13,17-seco-5α-androstan-17-oic-13,17-lactam, 3α-hydroxy-13α-amino-13,17-seco-5-androsten-17-oic-13,17-lactam and 17β-hydroxy-3-aza-A-homo-4α-androsten-4-one, have been prepared and their biological activity evaluated against P388 leukemia in vivo and Ehrlich Ascites tumor (EAT), P388 and L1210 leukemias and Baby Hamster cells (BHK) in vitro. The esters in which the alkylating congener is linked to the lactam alcohol in the axial position are inactive in vivo in P388 leukemia, while compounds 1, 4, 6, 13, 14 and the alkylating congeners 17, 18 and 20 are active. The effect of the homo-azasteroidal of N, N-bis(2-chloroethyl)aminobenzoic acid isomers and of 4-methyl-3-N, N-bis(2-chloroethyl)aminobenzoic acid on the incorporation of the radioactive precursor into the DNA of L1210, P388 leukemias, Ehrlich ascites tumor and, baby Hamster kidney cells was investigated. Higher inhibitory effects on the incorporation of the radioactive precursor was obtained with the ortho derivatives, yielding <70% inhibition of thymidine incorporation in all tumor lines tested.