897-06-3Relevant articles and documents
Preparation of androsta-1,4-diene-3,17-dione from sterols using Mycobacterium neoaurum VKPM Ac-1656 strain
Molchanova,Andryushina,Savinova,Stytsenko,Rodina,Voishvillo
, p. 354 - 358 (2007)
A product of microbiological cleavage of the sterols side chain, androsta-1,4-diene-3,17-dione, is toxic for bacteria, in particular, actinobacteria of the genera Mycobacterium and Arthrobacter. Sterols were transformed into androsta-1,4-diene-3,17-dione by culturing the M. neoaurum VKPM Ac-1656 strain in a high yield, provided that a sorbent was used for elimination of contact between the bacterial cells and the product. Unlike the cholesterol side chain, the more branched chains of phytosterols were cleaved in the presence of M. neoaurum at a high rate only under turbulent stirring of the culture medium, which intensified the formation of hydrocarbonate ion from NaNI3 in situ. Nauka/Interperiodica 2007.
Bioconversion of 4-androstene-3,17-dione to androst-1,4-diene-3,17-dione by recombinant Bacillus subtilis expressing ksdd gene encoding 3-ketosteroid-Δ1-dehydrogenase from Mycobacterium neoaurum JC-12
Zhang, Wenqing,Shao, Minglong,Rao, Zhiming,Xu, Meijuan,Zhang, Xian,Yang, Taowei,Li, Hui,Xu, Zhenghong
, p. 36 - 42 (2013)
The enzyme 3-ketosteroid-Δ1-dehydrogenase (KSDD), involved in steroid metabolism, catalyzes the transformation of 4-androstene-3,17-dione (AD) to androst-1,4-diene-3,17-dione (ADD) specifically. Its coding gene was obtained from Mycobacterium neoaurum JC-12 and expressed on the plasmid pMA5 in Bacillus subtilis 168. The successfully expressed KSDD was analyzed by native-PAGE. The activities of the recombinant enzyme in B. subtilis were 1.75 U/mg, which was about 5-fold that of the wild type in M. neoaurum. When using the whole-cells as catalysts, the products were analyzed by tin-layer chromatography and high-performance liquid chromatography. The recombinant B. subtilis catalyzed the biotransformation of AD to ADD in a percent conversion of 65.7% and showed about 18 folds higher than M. neoaurum JC-12. The time required for transformation of AD to ADD was about 10 h by the recombinant B. subtilis, much shorter than that of the wild-type strain and other reported strains. Thus, the efficiency of ADD production could be improved immensely. For industrial applications, the recombinant B. subtilis containing KSDD provides a new pathway of producing steroid medicines.
Synthesis of steroidal [1,2,4]triazolo[1,5-a]pyrimidines and their antiproliferative activities
Fan, Ning-Juan,Tang, Jiang-Jiang,Li, Yuan-Feng,Bai, Yu-Bin,Zhao, Xiao-Min
, p. 822 - 831 (2019/08/01)
— A facile strategy for the synthesis of steroidal [1,2,4]triazolo[1,5-a]pyrimidines 5a-g and 6a-g has been accomplished via a one-pot reaction of steroidal ketones, aromatic aldehydes and 3-amino-1,2,4-triazole in the presence of potassium tert-butoxide in refluxing tert-butanol. All the synthesized heterosteroids were evaluated for in vitro antiproliferative activity against human cancer cells by sulforhodamine B (SRB) assays. The preliminary results showed that compounds 6a and 6e possessed potent antiproliferative activities.
Synthesis of 3β-methyl ether of dehydroepiandrosterone by biotransformation of 3β-methyl ether of cholesterol with cells of mycobacteria Mycobacterium sp.
Andryushina,Stytsenko,Karpova,Yaderets,Zavarzin,Kurilov
, p. 2355 - 2358 (2020/02/18)
3p-Methyl ether of dehydroepiandrosterone was obtained by microbiological transformation of 3?-methyl ether of cholesterol with Mycobacterium sp. Androstane-3,17-dione, androst-4-ene-3,17-dione, and androsta-1,4-diene-3,17-dione were minor transformation products.
Hemisynthesis, computational and molecular docking studies of novel nitrogen containing steroidal aromatase inhibitors: Testolactam and testololactam
Lone, Shabir H.,Bhat, Muzzaffar A.,Lone, Rayees A.,Jameel, Salman,Lone, Javeed A.,Bhat, Khursheed A.
, p. 4579 - 4589 (2018/03/21)
Testololactone (10) and testolactone (11) represent aromatase inhibitors containing lactone rings. We previously reported their hemisynthesis from the most common phytosterols, which are highly abundant in nature. Herein, we report the synthesis of their nitrogen congeners: testololactam (3) and testolactam (8). The reaction process involves the conversion of 4-androstene-3,17-dione to its corresponding oxime using hydroxylamine hydrochloride, whose Beckmann rearrangement under acid conditions yielded the desired testololactam (3). However, testolactam (8) was formed by the Beckmann rearrangement of the oxime (7) of 1,4-androstene-3,17-dienone (6). This expeditious reaction scheme may be exploited for the bulk production of testololactam (3) and testolactam (8). Theoretical DFT studies concerning the structural and electronic properties of all the end products were carried out using the Becke three-parameter Lee-Yang-Parr function (B3LYP) and 6-31G(d,p) level of theory. Molecular electrostatic potential map and frontier orbital analysis were carried out. The HOMO-LUMO energy gap was calculated, which allowed the calculation of relative reactivity descriptors like chemical hardness, chemical inertness, chemical potential, nucleophilicity and electrophilicity index of the synthesized products. The molecular docking studies of testololactam (3), testolactam (8) and testololactone (10), with aromatase (CYP19) revealed binding free energies of (ΔGb) = -9.85, -9.62 and -10.14 kcal mol-1 respectively, compared to the standard testolactone (11), a well-known aromatase inhibitor sold under the brand name TESLAC, which exhibited a binding free energy (ΔGb) of -10.29 kcal mol-1 with an inhibition constant Ki of 28.87 nM. The docking study revealed that the nitrogen congeners exhibit a relatively lower but appreciable therapeutic efficiency to be used as aromatase inhibitors.
New product identification in the sterol metabolism by an industrial strain Mycobacterium neoaurum NRRL B-3805
Li, Xuemei,Chen, Xi,Wang, Yu,Yao, Peiyuan,Zhang, Rui,Feng, Jinhui,Wu, Qiaqing,Zhu, Dunming,Ma, Yanhe
supporting information, p. 40 - 45 (2018/02/21)
Mycobacterium neoaurum NRRL B-3805 metabolizes sterols to produce androst-4-en-3,17-dione (AD) as the main product, and androsta-1,4-dien-3,17-dione, 9α-hydroxy androst-4-en-3,17-dione and 22-hydroxy-23,24-bisnorchol-4-en-3-one have been identified as by-products. In this study, a new by-product was isolated from the metabolites of sterols and identified as methyl 3-oxo-23,24-bisnorchol-4-en-22-oate (BNC methyl ester), which was proposed to be produced via the esterification of BNC catalyzed by an O-methyltransferase using S-adenosyl-L-methionine as the methyl group donor. These results might open a new dimension for improvement of the efficiency of microbial AD production by eliminating this by-product via genetic manipulation of the strain.
Hemisynthesis of 2,3,4-13C3-1,4-androstadien-3,17-dione: A key precursor for the synthesis of 13C3-androstanes and 13C3-estranes
Berthonneau, Clément,Nun, Pierrick,Rivière, Matthieu,Pauvert, Mickael,Dénès, Fabrice,Lebreton, Jacques
, p. 3727 - 3737 (2018/04/14)
In this contribution, we describe two simple and efficient routes for the preparation of keto-aldehyde 1, a key intermediate for the synthesis of 13C3-androstanes and 13C3-estranes. In the first route, the targeted aldehyde 1 was obtained in 40% overall yield from 1,4-androstadien-3,17-dione (3 mmol scale) via a two-step sequence involving a one-pot, abnormal ozonolysis/sulfur oxidation/retro-Michael/ozonolysis process. Alternatively, a second route from 4-androsten-3,17-dione, using a six-step sequence, was optimized to produce 40 mmol batches of the key intermediate 1 in 42% overall yield. At the final stage, the A-ring was reconstructed through a Wittig reaction with the 1-triphenylphosphoranylidene-13C3-2-propanone 2, followed by an intramolecular condensation assisted by thioacetic acid via a Michael addition/retro-Michael reaction sequence to provide 2,3,4-13C3-1,4-androstadien-3,17-dione.
Microbial transformation of epiandrosterone by Aspergillus sydowii
Yildirim, Kudret,Kuru, Ali
, p. 718 - 721 (2016/12/30)
Incubation of epiandrosterone with Aspergillus sydowii MRC 200653 afforded ten metabolites. The fungal dehydrogenation of epiandrosterone is reported for the first time. The formation of the major metabolite, 6?-hydroxyandrost-4-ene-3,17-dione, involved first dehydrogenation to give a 4-ene and then hydroxylation at C-6?. Small amounts of the substrate were hydroxylated at C-1α, C-7α, C-7β and C-11α.
Phytosterols as precursors for the synthesis of aromatase inhibitors: Hemisynthesis of testololactone and testolactone
Lone, Shabir H.,Bhat, Khursheed A.
, p. 164 - 168 (2015/03/04)
Using β-sitosterol and stigmasterol as precursor materials, a concise and efficient hemisynthesis of aromatase inhibitors: testololactone and testolactone was accomplished in a well-established reaction scheme. It involves highly effective Oppaneur oxidation of both β-sitosterol as well as stigmasterol to generate the required enone moiety in ring 'A' of the desired steroid system. The Oppaneur oxidation products of both β-sitosterol and stigmasterol were then subjected to oxidative cleavage of the side chain to produce 4-androstene-3,17-dione. Baeyer-Villiger oxidation of 4-androstene-3,17-dione using m-CPBA yielded testololactone. Dehydrogenation of 4-androstene-3,17-dione using phenylselenyl chloride in ethyl acetate followed by selenoxide elimination with H2O2 in dichloromethane furnished androstenedienone. Baeyer-Villiger oxidation of the resulting androstenedienone yielded the desired testolactone (overall yield 33%). This expeditious reaction scheme may be exploited for the bulk production of aromatase inhibitors (especially testolactone marketed under the brand name Teslac) from the most abundant and naturally occurring phytosterols like β-sitosterol.
Structure and catalytic mechanism of 3-ketosteroid-Δ4-(5α)- dehydrogenase from Rhodococcus jostii RHA1 genome
Van Oosterwijk, Niels,Dijkstra, Bauke W.,Knol, Jan,Dijkhuizen, Lubbert,Van Der Geize, Robert
, p. 30975 - 30983,9 (2020/08/31)
3-Ketosteroid Δ4-(5α)-dehydrogenases (Δ4-(5α)- KSTDs) are enzymes that introduce a double bond between the C4 and C5 atoms of 3-keto-(5α)-steroids. Here we show that the ro05698 gene from Rhodococcus jostii RHA1 codes for a flavoprotein with Δ4-(5α)-KSTD activity. The 1.6 A resolution crystal structure of the enzyme revealed three conserved residues (Tyr-319, Tyr-466, and Ser-468) in a pocket near the isoalloxazine ring system of the FAD co-factor. Site-directed mutagenesis of these residues confirmed that they are absolutely essential for catalytic activity. A crystal structure with bound product 4-androstene-3,17-dione showed that Ser-468 is in a position in which it can serve as the base abstracting the 4β-proton from the C4 atom of the substrate. Ser-468 is assisted by Tyr-319, which possibly is involved in shuttling the proton to the solvent. Tyr-466 is at hydrogen bonding distance to the C3 oxygen atom of the substrate and can stabilize the keto-enol intermediate occurring during the reaction. Finally, the FAD N5 atom is in a position to be able to abstract the 5α-hydrogen of the substrate as a hydride ion. These features fully explain the reaction catalyzed by Δ4-(5α)-KSTDs.
