Microbial transformation of prednisone

The microbial transformation of prednisone (17alpha,21-dihydroxy-pregna-1,4-diene-3,11,20-trione) (1) by Cunninghamella elegans afforded two metabolites, 17alpha,21-dihydroxy-5alpha-pregn-1-ene-3,11,20-trione (2) and 17alpha,20S,21-trihydroxy-5alpha-pregn-1-ene-3,11-dione (3), while the fermentation of 1 with Fusarium lini, Rhizopus stolonifer and Curvularia lunata afforded a metabolite 1,4-pregnadiene-17alpha,20S,21-triol-3,11-dione (4). Compound 3 was found to be a new metabolite. Their structures were elucidated on the basis of spectroscopic techniques. Compound 3 showed inhibitory activity against lipoxygenase enzyme.     

Microbial transformation of (+)-androsta-1 ,4-diene-3,17-dione by Cephalosporium aphidicola

Fermentation of (+)-androsta-1,4-diene-3,17-dione ([structure: see text]) with Cephalosporium aphidicola for 8 days yielded oxidative and reductive metabolites, androst-4-ene-3,17-dione ([structure: see text]), 17beta-hydroxyandrosta-1,4-diene-3-one ([structure: see text]), 11alpha-hydroxyandrosta-1,4-diene-3,17-dione ([structure: see text]), 11alpha-hydroxyandrost-4-ene-3,17-dione ([structure: see text]), 11alpha,17beta-dihydroxyandrost-4-ene-3-one ([structure: see text]) and 11alpha,17beta-dihydroxyandrosta-1,4-diene-3-one ([structure: see text]). The fermentation of [structure: see text] with Fusarium lini also yielded metabolites [structure: see text]. The structures of these metabolites were elucidated on the basis of spectroscopic techniques.     

Biotransformation of (+)-androst-4-ene-3,17-dione

Fermentation of (+)-androst-4-ene-3,17-dione (1) with Curvularia lunata for 10 days yielded five oxidative and reductive metabolites, androsta-1,4-diene-3,17-dione (2), 17beta-hydroxyandrosta-1,4-dien-3-one (3), 11alpha-hydroxyandrost-4-ene-3,17-dione (4), 11alpha,17beta-dihydroxyandrost-4-en-3-one (5) and 15alpha-hydroxyandrosta-1,4-dien-17-one (6). The structures of these metabolites were elucidated on the basis of spectroscopic techniques. These microbially transformed products were assayed against the clinically important enzymes, tyrosinase and prolyl endopeptidase.     

Fractionation of free and conjugated steroids for the detection of boldenone metabolites in calf urine with ultra-performance liquid chromatography/tandem mass spectrometry

For over a decade there has been an intensive debate on the possible natural origin of boldenone (androst-1,4-diene-17beta-ol-3-one, 17beta-boldenone) in calf urine and several alternative markers to discriminate between endogenously formed boldenone and exogenously administered boldenone have been suggested. The currently approved method for proving illegal administration of beta-boldenone(ester) is the detection of beta-boldenone conjugates. In the presented method the sulphate, glucuronide and free fractions are separated from each other during cleanup on a SAX column to be able to determine the conjugated status of the boldenone metabolites. The sulphate and glucuronide fractions are submitted to hydrolysis and all three fractions are further cleaned up on a combination of C18/NH2 solid-phase extraction (SPE) columns. Chromatographic separation of the boldenone metabolites was achieved with a Waters Acquity UPLC instrument using a Sapphire C18 (1.7 microm; 2x50 mm) column within 5 min. Detection of the analytes was achieved by electrospray ionisation tandem mass spectrometry. The decision limits of this method, validated according to Commission Decision 2002/657/EC, were 0.08 ng mL(-1) for androsta-1,4-diene-3,17-dione, 0.13 ng mL(-1) for androst-4-ene-3,17-dione, 0.11 ng mL(-1) for 17alpha-boldenone, 0.07 ng mL(-1) for 17beta-boldenone, 0.24 ng mL(-1) for 5beta-androst-1-en-17beta-ol-3-one and 0.58 ng mL(-1) for 6beta-hydroxy-17beta-boldenone. Because of the fractionation approach used in this method there is no need for conjugated reference standards which often are not available. The disadvantage of needing three analytical runs to determine the conjugated status of each of the metabolites was overcome by using fast chromatography.     

Photochemistry of desonide, a non-fluorinated steroidal anti-inflammatory drug

The photochemistry of anti-inflammatory drug desonide (De, 1) was studied in aerobic as well as in anaerobic condition with different irradiation wavelengths (254, 310 nm) in acetonitrile and 2-propanol. All photoproducts obtained were isolated and characterized on the basis of IR, (1)H-, (13)C-NMR spectroscopy and elemental analysis study. The products were: 11beta,21-dihydroxy-16alpha,17alpha-(1-methylethylidenedioxy)-1,5-cyclopregn-3-ene-2,20-dione 2 (254 nm), 11beta-hydroxy-16alpha,17alpha-(1-methylethylidenedioxy)androsta-1,4-diene-3-one 3 (310 nm/2-propanol), 17beta-hydroperoxy-11beta-hydroxy-16alpha,17alpha-(1-methylethylidenedioxy)androsta-1,4-diene-3-one 4 (310 nm/O(2)/2-propanol). Cyclohexadienone moiety in ring A and keto group at C(17) were found to be deeply modified by UV light therefore, loss of biological activity both during storage and in vivo can not be ruled out.     

Synthesis of α,ω-bis(pyrazol-3-yl)alkanes: I. 1,4-bis(1-methyl- and 1-benzyl-5-chloro-1<Emphasis Type="Italic">H</Emphasis>-pyrazol-3-yl)-butanes from 1,1,10,10-Tetrachlorodeca-1,9-diene-3,8-dione and 1,1-dimethyl- or benzylhydrazine

First representatives of bis-2-chloro- and 2,2-dichlorovinyl ketones, 1,10-dichlorodeca-1,9-diene-3,8-dione and 1,1,10,10-tetrachlorodeca-1,9-diene-3,8-dione, were synthesized by reaction of hexanedioyl dichloride with acetylene and 1,1-dichloroethene, respectively, in the presence of AlCl₃. 1,1,10,10-Tetrachlorodeca-1,9-diene-3,8-dione reacted with benzylhydrazine and 1,1-dimethylhydrazine to give 1,4-bis(1-benzyl-5-chloro-1H-pyrazol-3-yl)butane and 1,4-bis(5-chloro-1-methyl-1H-pyrazol-3-yl)butane, respectively.     

Microbial transformation of dehydroepiandrosterone

Transformation of dehydroepiandrosterone (DHEA) (1) was carried out by a plant pathogen Rhizopus stolonifer, which resulted in the production of seven metabolites. These metabolites were identified as 3beta,17beta-dihydroxyanandrost-5-ene (2), 3beta,17beta-dihydroxyandrost-4ene (3), 17beta-hydroxyandrost-4-ene-3-one (4), 3beta,11-dihydroxyandrost-4-ene-17-one (5), 3beta,7alpha-dihydroandrost-5-ene-17-one (6), 3A,7alpha,17beta-trihydroxyandrost-5-ene (7) and 11beta-hydroxyandrost-4,6-diene-3,17-dione (8). The structures of the transformed products were determined by the spectroscopic techniques.     

Microbial hydroxylation of pregnenolone derivatives

Pregnenolone and pregnenolone acetate were incubated with the fungi Cunninghamella elegans, Rhizopus stolonifer and Gibberella fujikuroi. Incubation of with C. elegans yielded metabolites, 3beta,7beta,11alpha-trihydroxypreg-5-en-20-one, 3beta,6alpha,11alpha,12beta,15beta-pentahydroxypreg-4-en-20-one and 3beta,6beta,11alpha-trihydroxypreg-4-en-20-one, while incubation with G. fujikuroi yielded two known metabolites, 3beta,7beta-dihydroxypregn-5-en-20-one and 6beta,15beta-dihydroxypreg-4-ene-3,20-dione. Metabolites and were found to be new. Fermentation of by C. elegans yielded four known oxidative metabolites, androsta-1,4-diene-3,17-dione, 6beta,15beta-dihydroxyandrost-4-ene-3,17-dione and 11alpha,15beta-dihydroxypreg-4-ene-3,20-dione. Fermentation of with R. stolonifer yielded two known metabolites, 11alpha-hydroxypreg-4-ene-3,20-dione and. Compounds were screened for their cholinesterase inhibitory activity in a mechanism-based assay.     

Effect of structure on the electron-capture negative ionization mass spectrometric response of steroids

The key to enhanced electron-capture negative ion mass spectrometry (ECNI-MS) response of ketosteroids is extensive α,β-unsaturation. Combinations of double bonds, carbonyl groups, epoxides and halogens within a steroid nucleus were correlated with their ECNI responses. The greatest ECNI responses result from extensive conjugation, such as the linearly conjugated 4-ene-3,6-dione system and the cross-conjugated 1,4-diene-3,11-dione system. In general, an epoxy ketosteroid has a relative response 40 times greater than that of its corresponding enone. In an α-halo ketone, an axial halogen substitution will increase the relative response by an order of magnitude, but an equatorial halogen substituent will not affect the response. The fragmentation of ketosteroids as related to ECNI response is discussed.     

A new bibenzyl derivative from Pleione bulbocodioides

A new bibenzyl derivative 1, named 2-（4＇＇-hydroxybenzy1）-3-（3＇-hydroxyphenethy1）-5-methoxy-cyclohexa-2,5-diene-1,4- dione, and two known stilbenoids （2, 3） were isolated from the tubers of Pleione bulbocodioides （Franch.） Rolfe. Their structures were elucidated by spectroscopic methods.     

Biotransformation of phorbol by human intestinal bacteria

Anaerobic incubation of phorbol (1) from Croton tiglium with human intestinal bacteria afforded five metabolites: isophorbol (2), deoxyphorbol (3), 4beta,9alpha,20-trihydroxy-13,15-seco-1,6,15-tigliatriene-3,13-dione (4), 4beta,9alpha,20-trihydroxy-15,16,17-trinor-1,6-tigliadiene-3,13-dione (5) and 4beta,9a,20-trihydroxy-14(13-->12)-abeo-12alphaH-1,6-tigliadiene-3,13-dione (6). All these metabolites (2-6) were identified and characterized by spectroscopic means, including two-dimensional (2D)-NMR. Nine defined strains from the human intestine showed an ability to transform 1 to these metabolites.     

A new bibenzyl derivative from Pleione bulbocodioides

A new bibenzyl derivative 1,named 2-(4"-hydroxybenzyl)-3-(3'-hydroxyphenethyl)-5-methoxy-cyclohexa-2,5-diene-1,4- dione,and two known stilbenoids(2,3)were isolated from the tubers of Pleione bulbocodioides(Franch.)Rolfe.Their structures were elucidated by spectroscopic methods.     

A Green Blue LED-Driven Two-Liquid-Phase One-Pot Procedure for the Synthesis of Estrogen-Related Quinol Prodrugs

Quinol derivatives of estrogens are effective pro-drugs in steroid replacement therapy. Here, we report that these compounds can be synthesized in one-pot conditions and high yield by blue LED-driven photo-oxygenation of parent estrogens. The oxidation was performed in buffer and eco-certified 2-methyltetrahydrofuran as the two-liquid-phase reaction solvent, and in the presence of meso-tetraphenyl porphyrin as the photosensitizer. Two steroidal prodrugs 10β, 17β-dihydroxyestra-1,4-dien-3-one (DHED) and 10β-Hydroxyestra-1,4-diene-3,17-dione (HEDD) were obtained with high yield and selectivity.     

Rearrangement of 1,4-benzodiazepin-2,4-diones to 3,l-benzoxazin-4-ones

Treatment of 7-chloro-3,4-dihydro-1H-1,4-benzodiazepin-2,5-dione (Ia) with refluxing acetic anhydride in the presence of pyridine afforded 6-chloro-2-methyl-4H-3,1-benzoxazin-4-one (IIa). A plausible reaction path for this novel rearrangement reaction is described: Ia → 4-acetyl-7-chloro-3,4-dihydro-lH-1,4-benzodiazepin-2,5-dione → 7-chloro-1,4-diacetyl-3,4-dihydro-lH-1,4-benzodiazepin-2,4-dione → IIa. When 7-chloro-3,4-dihydro-4-methyl-lH-1,4-benzodiazepin-2,5-dione (Ib), 3,4-dihydro-4-methyl-1H-1,4-benzodiazepin-2,5-dione (Id) and 3,4-dihydro-1-methyl-1H-1,4-benzodiazepin-2,5-dione (Ie) were allowed to react with acetic anhydride under conditions similar to those used for the rearrangement reaction, only acetylation occurred.     

Structure elucidation and NMR assignments for two amide alkaloids from a Mangrove endophytic Fungus (No. ZZF-22)

The structure elucidations and complete (1)H and (13)C NMR assignments are reported for two new natural products: 3-benzylidene-8,8a-dihydroxy-2-methyl-hexahydro-pyrrolo[1,2-a]pyrazine-1,4-dione(1) and 4-hydroxy-6-(hydroxy-phenyl-methyl)-N-(3-methyl-butyryl)-nicotinamide (2). Both of these secondary metabolites were isolated from the fermentation medium of a Mangrove endophytic fungus. High resolution electron impact mass spectrometry (HREIMS), FT-IR Spectroscopy and NMR experiments including gCOSY, gHMQC, gHMBC and NOE were used for determination of the structures and assignments of the amide alkaloids.     

Triterpenes from the spores of Ganoderma lucidum and their cytotoxicity against meth-A and LLC tumor cells

Six new highly oxygenated lanostane-type triterpenes, called ganoderic acid gamma (1), ganoderic acid delta (2), ganoderic acid epsilon (3), ganoderic acid zeta (4), ganoderic acid eta (5) and ganoderic acid theta (6), were isolated from the spores of Ganoderma lucidum, together with known ganolucidic acid D (7) and ganoderic acid C2 (8). Their structures of the new triterpenes were determined as (23S)-7beta,15alpha,23-trihydroxy-3,11-dioxolanosta-8, 24(E)-diene-26-oic acid (1), (23S)-7alpha,15alpha23-trihydroxy-3,11-dioxolanosta-8, 24(E)-diene-26-oic acid (2), (23S)-3beta3,7beta, 23-trihydroxy-11,15-dioxolanosta-8,24(E)-diene-26-oic acid (3), (23S)-3beta,23-dihydroxy-7,11,15-trioxolanosta-8, 24(E)-diene-26-oic acid (4), (23S)-3beta,7beta,12beta,23-tetrahydroxy-11,15-dioxolanos ta-8,24(E)-diene-26-oic acid (5) and (23S)-3beta,12beta23-trihydroxy-7,11,15-trioxolanosta-8,24(E )-diene-26-oic acid (6), respectively, by chemical and spectroscopic means, which included the determination of a chiral center in the side chain by a modification of Mosher's method. The cytotoxicity of the compounds isolated from the Ganoderma spores was carried out in vitro against Meth-A and LLC tumor cell lines.     

The quinones of bicyclo[3.1.0]hexatriene: A computational study of their chemistry and thermochemistry

Quinones of bicyclo[3.1.0]hexa-1,3,5-triene were examined computationally. The six compounds considered were the five possible classical and one non-classical quinone: bicyclo[3.1.0]hexa-1(6),4-diene-2,3-dione (and its monocyclic isomer with a long trans-annular bond), bicyclo[3.1.0]hexa-1(5),3-diene-2,6-dione, bicyclo[3.1.0]hexa-1,4-diene-3,6-dione (and its monocyclic isomer with a long trans-annular bond), and bicyclo[3.1.0]hexa-1(5),4-diene-2,4-dione-3,6-diyl, a non-classical (non-Kekulé) zwitterion. The two long trans-annular bond structures are akin to that found for m-benzyne. Geometries were calculated (BLYP/6-31G¹, CASSCF(2,2)/6-31G¹, MP2/6-31G¹) and electronic structural inferences were made from the geometries. Also calculated were relative energies and heats of formation (CBS-QB3), singlet and triplet energies (BLYP/6-31G¹), and ionization energies and electron affinities (HF/6-311+G⁷//BLYP/6-31G¹). The NICS(1) calculations were performed as a probe of the aromaticity of the diverse quinones.     

Epoxidation and reduction of DHEA, 1,4,6-androstatrien-3-one and 4,6-androstadien-3beta,17beta-diol

Dehydroepiandrosterone (DHEA) reacted with m-chloroperoxybenzoic acid(m-CPBA) to form 3beta-hydroxy-5alpha,6alpha-epoxyandrostan-17-one (1), but it did not react with 30% H2O2. 1,4,6-Androstatrien-3,17-dione (2) was obtained from DHEA and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone in dioxane. Compound 2 was reacted with 30%H2O2 and 5% NaOH in methanol to give 1alpha,2alpha-epoxy-4,6-androstadien-3,17-dione (3),which was stereoselectively reduced with NaBH4 to form 1alpha,2alpha-epoxy-4,6-androstadien-3beta,17beta-diol (7) and reacted with Li metal in absolute ethanol-tetrahydrofuran mixture to give 2-ethoxy-1,4,6-androstatrien-3,17-dione (8). Compound 2 was also epoxidized with m-CPBA in dichloromethane to afford 6alpha,7alpha-epoxy-1,4-androstadien-3,17-dione (4),which was reacted with NaBH4 to synthesize 6alpha,7alpha-epoxy-4-androsten-3beta,17beta-diol (9).Compound 4 was reduced with Li metal in absolute ethanol-tetrahydrofuran mixture to form 7beta-ethoxy-6alpha-hydroxy-1,4-androstadien-3,17-dione (10). Compound 2 was reduced with NaBH4 in absolute ethanol to form 4,6-androstadien-3beta,17beta-diol (5), which was reacted with 30% H2O2 to give the original compound, but which reacted with m-CPBAto give 4beta,5beta-epoxy-6-androsten-3beta,17beta-diol (6).     

Synthesis and Biological Activity of Cyanoethyl Derivatives of Fusidic Acid

3,11-Dihydroxy and 3,11-dioxo triterpenoids of the fusidane series reacted with acrylonitrile in 1,4-dioxane in the presence of alkali and phase-transfer catalyst to give mono- and bis(2-cyanoethoxy) and 2-cyanoethyl derivatives. The reaction with 3,11-dioxo analog afforded 2,2-disubstituted derivative as a result of addition of two cyanoethyl groups to the α-position with respect to the C³=O carbonyl group. The isolated compounds were screened for antibacterial and antifungal activities.

     

Reaction of dicobalt octacarbonyl with acetylene and carbon monoxide at low temperature and pressure: Formation of cyclic enone products

Dicobalt octacarbonyl is shown to react with acetylene and carbon monoxide under mild conditions in dimethoxyethane or benzene to produce, in low yields, bicyclo[3.3.0]octa-3,7-diene-2,6-dione, benzoquinone, and the cyclopentadienone-derived products 3a,4,7,7a-tetrahydro-2,7-methanoindene-1,10-dione, 1-indanone, tetracyclo[5.5.2.0^2,60^8,12]tetradeca-4,10,13-triene-3,9-dione, and tetracyclo[5.5.2.0^2,60^8,12]tetradeca-4,9,13-triene-3,11-dione. Possible mechanisms for the formation of these products are discussed.