- Evaluation of the fluorescent probes Nile Red and 25-NBD-cholesterol as substrates for steroid-converting oxidoreductases using pure enzymes and microorganisms
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The fluorescent probes Nile Red (nonsteroidal dye) and 25-{N-[(7-nitrobenz- 2-oxa-1,3-diazol-4-yl)-methyl]amino}-27-norcholesterol (25-NBD-cholesterol) (a cholesterol analog) were evaluated as novel substrates for steroid-converting oxidoreductases. Docking simulations with autodock showed that Nile Red fits well into the substrate-binding site of cytochrome P450 17α-hydroxylase/ 17,20-lyase (CYP17A1) (binding energy value of -8.3 kcal·mol -1). Recombinant Saccharomyces cerevisiae and Yarrowia lipolytica, both expressing CYP17A1, were found to catalyze the conversion of Nile Red into two N-dealkylated derivatives. The conversion by the yeasts was shown to increase in the cases of coexpression of electron-donating partners of CYP17A1. The highest specific activity value (1.30 ± 0.02 min-1) was achieved for the strain Y. lipolytica DC5, expressing CYP17A1 and the yeast's NADPH-cytochrome P450 reductase. The dye was also metabolized by pure CYP17A1 into the N-dealkylated derivatives, and gave a type I difference spectrum when titrated into low-spin CYP17A1. Analogously, docking simulations demonstrated that 25-NBD-cholesterol binds into the active site of the microbial cholesterol oxidase (CHOX) from Brevibacterium sterolicum (binding energy value of -5.6 kcal·mol-1). The steroid was found to be converted into its 4-en-3-one derivative by CHOX (Km and kcat values were estimated to be 58.1 ± 5.9 μm and 0.66 ± 0.14 s-1, respectively). The 4-en-3-one derivative was also detected as the product of 25-NBD-cholesterol oxidation with both pure microbial cholesterol dehydrogenase (CHDH) and a pathogenic bacterium, Pseudomonas aeruginosa, possessing CHOXs and CHDHs. These results provide novel opportunities for investigation of the structure-function relationships of the aforementioned oxidoreductases, which catalyze essential steps of steroid bioconversion in mammals (CYP17A1) and bacteria (CHOX and CHDH), with fluorescence-based techniques. Docking simulations show that fluorescent substances Nile Red and 25-NBD-cholesterol fit well the substrate-binding sites of CYP17A1 and the cholesterol oxidase (CHOX), respectively. Recombinant yeasts, expressing CYP17A1, as well as pure CYP17A1 catalyze the Nile Red conversion into N-dealkylated derivatives. 25-NBD-cholesterol is converted into its 4-en-3-one derivative by the CHOX and cholesterol dehydrogenase as well as by bacteria Pseudomonas aeruginosa.
- Faletrov, Yaroslav V.,Frolova, Nina S.,Hlushko, Hanna V.,Rudaya, Elena V.,Edimecheva, Irina P.,Mauersberger, Stephan,Shkumatov, Vladimir M.
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Read Online
- Modified bile acids and androstanes—Novel promising inhibitors of human cytochrome P450 17A1
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Cytochromes P450 are key enzymes for steroid hormone biosynthesis in human body. They are considered as targets for the screening of novel high efficient drugs. The results of screening of bile acids and androstane derivatives toward human recombinant steroid 17α-hydroxylase/17,20-lyase (CYP17A1) are presented in this paper. A group of steroids, binding with micromolar or submicromolar affinity (in a range from 9 μM – less than 0.1 μM), was identified. Results presented here showed that these steroidal compounds are able to decrease rate of hydroxylation of essential CYP17A1 substrate – progesterone, while some compounds completely inhibited enzyme activity. Structure-activity relationship (SAR) analysis based on in vitro and in silico studies showed that high affinity of the enzyme to bile acids derivatives is correlated with side chain hydrophobicity and presence of hydroxyl or keto group at C3 position. From the other side, bile acid-derived compounds with more polar side chain or substituents at C7 and C12 positions possess higher Kd values. Among androstane-derived steroids couple of Δ5-steroids with hydroxyl group at C3 position, as well as 16,17-secosteroids, were found to be high affinity ligands of this enzyme. The data obtained could be useful for the design of novel highly efficient inhibitors of CYP17A1, since the bile acids-derived compounds are for first time recognized as effective CYP17A1 inhibitors.
- Dzichenka, Yaraslau,Shapira, Michail,Yantsevich, Aliaksei,Cherkesova, Tatsiana,Grbovi?, Ljubica,Savi?, Marina,Usanov, Sergey,Jovanovi?-?anta, Suzana
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- Inherent steroid 17α,20-lyase activity in defunct cytochrome P450 17A enzymes
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Cytochrome P450 (P450) 17A1 catalyzes the oxidations of progesterone and pregnenolone and is the major source of androgens. The enzyme catalyzes both 17α-hydroxylation and a subsequent 17α,20-lyase reaction, and several mechanisms have been proposed for the latter step. Zebrafish P450 17A2 catalyzes only the 17α-hydroxylations. We previously reported high similarity of the crystal structures of zebrafish P450 17A1 and 17A2 and human P450 17A1. Five residues near the heme, which differed, were changed. We also crystallized this five-residue zebrafish P450 17A1 mutant, and the active site still resembled the structure in the other proteins, with some important differences. These P450 17A1 and 17A2 mutants had catalytic profiles more similar to each other than did the wildtype proteins. Docking with these structures can explain several minor products, which require multiple enzyme conformations. The 17α-hydroperoxy (OOH) derivatives of the steroids were used as oxygen surrogates. Human P450 17A1 and zebrafish P450s 17A1 and P450 17A2 readily converted these to the lyase products in the absence of other proteins or cofactors (with catalytically competent kinetics) plus hydroxylated 17α-hydroxysteroids. The 17α-OOH results indicate that a "Compound I" (FeO3+) intermediate is capable of formation and can be used to rationalize the products. We conclude that zebrafish P450 17A2 is capable of lyase activity with the 17α-OOH steroids because it can achieve an appropriate conformation for lyase catalysis in this system that is precluded in the conventional reaction.
- Gonzalez, Eric,Johnson, Kevin M.,Pallan, Pradeep S.,Phan, Thanh T.N.,Zhang, Wei,Lei, Li,Wawrzak, Zdzislaw,Yoshimoto, Francis K.,Egli, Martin,Peter Guengerich
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p. 541 - 556
(2018/02/14)
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- A 17a - hydroxy progesterone preparation method
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The invention relates to a method for preparing 17alpha-hydroxyprogesterone. The17alpha-hydroxyprogesterone is prepared by taking 17beta- cyano-5-androstene-17-ol-3,3-diethylene ketal (referred as an intermediate II) as a raw material and dimethylzinc or methylzinc chloride as a reagent; the content of the 17alpha-hydroxyprogesterone by HPLC is above 99.5% and the weight yield is 83-87%. The method comprises the following steps of dissolving the intermediate II in an organic solvent, adding lithium chloride as a catalyst, stirring, raising the temperature to 40-80 DEG C, dropwise adding a toluene solution of dimethylzinc or methylzinc chloride of which the concentration is 2M, and continuing to complete the reaction; and then adding an ammonium chloride solution of which the concentration is 25% to destruct an organic zinc reagent, separating the aqueous layer out and extracting, merging the organic layer and the extract and concentrating the solvent to near dryness, and then adding lower alcohol, stirring, raising the temperature to 40-60 DEG C, adding the acid of which the concentration is 2M, hydrolyzing, adjusting the pH value with a weak base after the reaction is completed, evaporating 90% of the solvent out, adding tap water, cooling and crystallizing to obtain a crude 17alpha-hydroxyprogesterone product; and then carrying out reflux decolorizing on the crude product with activated carbon by virtue of alcohol, and refining to obtain the commercial grade 17alpha-hydroxyprogesterone. The 17alpha-hydroxyprogesterone produced by the method disclosed by the invention has the advantages of good purity and high yield and is economic and environment-friendly, and the solvent can be recycled.
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Paragraph 0017; 0018; 0019; 0020; 0021; 0022; 0023; 0024
(2017/08/25)
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- Method for preparing 17alpha-hydroxyprogesterone
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The invention discloses a method for preparing 17alpha-hydroxyprogesterone. According to the method, 4-androstenedione I serves as the raw material, a 17-site branched chain is introduced through ethynylation reaction, a 17-site beta-hydroxyl group is converted into an alpha-hydroxyl group through catalytic reaction, and finally 17alpha-hydroxyprogesterone is prepared through carbonylation reaction. The reaction formula is shown in the description. Four steps of reaction are conducted with 4-androstenedione as the raw material, a 17-site side chain is introduced through ethynylation reaction, the 17beta-hydroxyl group is converted into the 17alpha-hydroxyl group through transposition reaction, the weight yield of each step ranges from 85% to 109%, the total weigh yield is 85% or above, use of virulent acetone cyanohydrin is avoided, the raw material conversion rate is high, selectivity is good, production cost of 17alpha-hydroxyprogesterone is reduced, the process is safe, and large-scale industrial production is easy.
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- A high yield of 17 α - hydroxy progesterone of simple preparation method
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The invention relates to a high-yield simple preparation method of 17alpha-hydroxy progesterone. With 4-androstenedione as an initial raw material, the method comprises the following steps: performing a cyanogen alcoholization addition reaction between the 17-site carbonyl of 4-androstenedione and acetone hydrogen alcohol to obtain 17-alpha hydroxyl-17-beta cyanoandrostane-4-ene-3-one; performing a ketal protection reaction of the C3-site carbonyl to obtain a ketal product; and performing a direct methylation reaction between the ketal product and zinc chloride methane, and hydrolyzing to obtain 17alpha-hydroxy progesterone. The method provided by the invention has the advantages of short process, high yield, high product purity, mild reaction conditions, low cost and low energy consumption and is particularly suitable for industrial production.
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Paragraph 0031; 0034; 0035; 0036; 0037; 0038; 0039; 0040
(2017/11/18)
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- Preparation method of steroidal compound with multiple olefin groups
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The invention relates to a preparation method of a steroid medicine intermediate body, in particular to a method for preparing a steroid carrying medicine intermediate body, namely steroid carrying-1,4,9(11),16(17)-tetraterpene-3,20-diketone and steroid carrying-4,9(11)-diene-3,20-diketone-17 alpha-hydroxyl, by taking androstane-4-alkene-3,17-diketone as a substrate.
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Paragraph 0091-0092; 0096-0098
(2017/11/04)
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- A synthetic 17 α-hydroxy progesterone of the new method
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The invention provides a novel method of synthesizing 17 alpha-hydroxyl progesterone (01). The novel method comprises the following steps: 1) in the presence of trimethylsilyl chloride and a methyl magnesium chloride solution, performing double-protection reaction on 17 bata-cyano-17 alpha-hydroxyl-4-androstene-3-one (02) to produce a compound (03); 2) heating up to perform grignard addition reaction, and transferring into an ammonium chloride aqueous solution to produce a compound (04); 3) preserving the heat and discoloring by active carbon, adding diluted acid water to reflux and perform hydrolysis reaction of double protection groups, and separating and purifying to obtain 17 alpha-hydroxyl progesterone (01). According to the novel method disclosed by the invention, the conventional 3-position ketal protection, 17 alpha-hydroxyl vinyl ether protection, grignard addition, re-hydration of protection groups on position 3 and position 17 and discoloring-refining are simplified into one-step operation.
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Paragraph 0038; 0039
(2017/02/28)
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- A by the 3,17-dione steroid preparing steroid the synthetic method of the compound of
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The invention discloses a synthetic method of steroid type drugs and intermediates, in particular relates to a synthetic method for preparing 17-hydroxy-20-ketone steroid compounds from 3,17-diketone steroids, and belongs to the field of synthesis of drugs. The method takes 3,17-diketone steroids as raw materials and adopts a conventional, environment-friendly, low-toxicity reagent, and the steroid type drugs such as cortisone, hydrocortisone, metacortandracin, or hydroprednisone, or intermediates 17alpha-hydroxy-20-ketone compounds are prepared simply and conveniently at high yield by selective protection of C3 or (and) C11-ketone group, Wittig reaction of C17, selective oxidization of 17(20)-position double bonds and halogenating replacement. The aftertreatment is simple, few three wastes are generated, the reaction selectivity is good, the yield is high, and byproducts anti-pregnancy steroidal drugs and steroidal compounds can be obtained. The raw materials are easy to get, the cost is low and the synthetic process is simple; the synthetic method is suitable for industrial production.
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- Synthesis of [3α-3H] 17α-Hydroxy pregnenolone and [3α-3H] Pregnenolone
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For the first time, [3α-3H] 17α-hydroxy pregnenolone (1) was synthesized through a multiple step sequence. The presence of [3β-3H] isomer in RP-HPLC purified product was identified by tritium NMR. The [3β-3H] isomer was then separated from [3α-3H] 17α-hydroxy pregnenolone with chiralPAK AD-H column. [3α-3H] pregnenolone (2) was synthesized from commercial available 5-pregnen-3,20-dione in one step with an improved procedure. [3α-3H] 17α-hydroxy pregnenolone (1) was synthesized through a multiple step sequence. The presence of [3β- 3H] isomer was identified then separated with chiralPAK AD-H column. [3α-3H] pregnenolone (2) was synthesized from commercial available 5-pregnen-3,20-dione in one step. Copyright
- Tian, Yuan,Hong, Yang,Bonacorsi, Samuel J.,Balog, Aaron,Gong, Sharon
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- Double site saturation mutagenesis of the human cytochrome P450 2D6 results in regioselective steroid hydroxylation
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The human cytochrome P450 2D6 (CYP2D6) is one of the major human drug metabolizing enzymes and acts preferably on substrates containing a basic nitrogen atom. Testosterone - just as other steroids - is an atypical substrate and only poorly metabolized by CYP2D6. The present study intended to investigate the influence of the two active site residues 216 and 483 on the capability of CYP2D6 to hydroxylate steroids such as for example testosterone. All 400 possible combinatorial mutations at these two positions have been generated and expressed individually in Pichia pastoris. Employing whole-cell biotransformations coupled with HPLC-MS analysis the testosterone hydroxylase activity and regioselectivity of every single CYP2D6 variant was determined. Covering the whole sequence space, CYP2D6 variants with improved activity and so far unknown regio-preference in testosterone hydroxylation were identified. Most intriguingly and in contrast to previous literature reports about mutein F483I, the mutation F483G led to preferred hydroxylation at the 2β-position, while the slow formation of 6β-hydroxytestosterone, the main product of wild-type CYP2D6, was further reduced. Two point mutations have already been sufficient to convert CYP2D6 into a steroid hydroxylase with the highest ever reported testosterone hydroxylation rate for this enzyme, which is of the same order of magnitude as for the conversion of the standard substrate bufuralol by wild-type CYP2D6. Furthermore, this study is also an example for efficient human CYP engineering in P. pastoris for biocatalytic applications and to study so far unknown pharmacokinetic effects of individual and combined mutations in these key enzymes of the human drug metabolism. 400 cytochrome P450 2D6 (CYP2D6) variants representing all possible amino acid exchanges at two important enzyme's residues were expressed and individually analyzed to investigate their influence on regioselective steroid hydroxylation. Steroids represent a substrate class atypical for wildtype CYP2D6. Employing this strategy CYP2D6 variants with improved activity and variants with altered region-preference were identified and characterized.
- Geier, Martina,Braun, Andreas,Fladischer, Patrik,Stepniak, Piotr,Rudroff, Florian,Hametner, Christian,Mihovilovic, Marko D.,Glieder, Anton
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p. 3094 - 3108
(2013/07/26)
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- Rabbit 3-hydroxyhexobarbital dehydrogenase is a NADPH-preferring reductase with broad substrate specificity for ketosteroids, prostaglandin D2, and other endogenous and xenobiotic carbonyl compounds
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3-Hydroxyhexobarbital dehydrogenase (3HBD) catalyzes NAD(P) +-linked oxidation of 3-hydroxyhexobarbital into 3-oxohexobarbital. The enzyme has been thought to act as a dehydrogenase for xenobiotic alcohols and some hydroxysteroids, but its physiological function remains unknown. We have purified rabbit 3HBD, isolated its cDNA, and examined its specificity for coenzymes and substrates, reaction directionality and tissue distribution. 3HBD is a member (AKR1C29) of the aldo-keto reductase (AKR) superfamily, and exhibited high preference for NADP(H) over NAD(H) at a physiological pH of 7.4. In the NADPH-linked reduction, 3HBD showed broad substrate specificity for a variety of quinones, ketones and aldehydes, including 3-, 17- and 20-ketosteroids and prostaglandin D2, which were converted to 3α-, 17β- and 20α-hydroxysteroids and 9α,11β- prostaglandin F2, respectively. Especially, α-diketones (such as isatin and diacetyl) and lipid peroxidation-derived aldehydes (such as 4-oxo- and 4-hydroxy-2-nonenals) were excellent substrates showing low Km values (0.1-5.9 μM). In 3HBD-overexpressed cells, 3-oxohexobarbital and 5β-androstan-3α-ol-17-one were metabolized into 3-hydroxyhexobarbital and 5β-androstane-3α,17β-diol, respectively, but the reverse reactions did not proceed. The overexpression of the enzyme in the cells decreased the cytotoxicity of 4-oxo-2-nonenal. The mRNA for 3HBD was ubiquitously expressed in rabbit tissues. The results suggest that 3HBD is an NADPH-preferring reductase, and plays roles in the metabolisms of steroids, prostaglandin D2, carbohydrates and xenobiotics, as well as a defense system, protecting against reactive carbonyl compounds.
- Endo, Satoshi,Matsunaga, Toshiyuki,Matsumoto, Atsuko,Arai, Yuki,Ohno, Satoshi,El-Kabbani, Ossama,Tajima, Kazuo,Bunai, Yasuo,Yamano, Shigeru,Hara, Akira,Kitade, Yukio
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p. 1366 - 1375
(2013/11/19)
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- A direct stereoselective preparation of a fish pheromone and application of the zinc porphyrin tweezer chiroptical protocol in its stereochemical assignment
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A two-step stereoselective preparation of a goldfish pheromone, 17α,20β-dihydroxy-4-pregnen-3-one, is reported from the readily available cortexolone in 64% overall yield. The (20S)-epimer was also synthesized in three steps from cortexolone with an overall yield of 47%. A microscale chiroptical technique based on a host/guest complexation mechanism between the substrate and a dimeric metalloporphyrin host (tweezer) was used to confirm the stereochemical assignment, while Density Functional Theory (DFT) calculations were employed to explain the high stereoselectivity induced by the 17α-hydroxyl and C18-methyl groups. Chirality 00:000-000:, 2013. 2013 Wiley Periodicals, Inc.
- Ouedraogo, Yannick P.,Huang, Longchuan,Torrente, Mariana P.,Proni, Gloria,Chadwick, Ekaterina,Wehmschulte, Rudolf J.,Nesnas, Nasri
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p. 575 - 581
(2013/09/12)
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- Molecular interactions of progesterone derivatives with 5α-reductase types 1 and 2 and androgen receptors
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The aim of this study was to ascertain the inhibitory effect of several progesterone derivatives for 5α-reductase types 1 and 2 isozymes and to determine the binding to the androgen receptor. The 3,20-dioxopregna-4-ene-17α-yl acetate 4 containing an acetoxy group in C-17 and steroid 17α-hydroxypregn-4-ene-3,20-dione 5 having a hydroxyl group in the same position inhibited both isozymes. On the other hand, 17α-hydroxy-4,5-epoxypregnan-3,20-dione 6 with an epoxy function at C-4, inhibited only the type 1 enzyme. Steroid 4-chloro-17α-hydroxypregn-4-ene-3,20-dione 7a and 4-bromo-17α-hydroxypregn-4-ene-3,20-dione 7b having the C-4 conjugated system and a chlorine or a bromine atom at C-4 respectively, inhibited both types of 5α-reductase. These results indicate that an increase in the electronegativity of ring A produces a major inhibitory activity for 5α-reductase type 1; however this increase was not observed for type 2 enzyme. When the free hydroxyl group of 7a or 7b was esterified, compounds 3,20-dioxo-4-chloropregn-4-ene-17α yl-4-ethylbenzoate 8a and 3,20-dioxo-4-bromopregn-4-ene-17α yl-4-ethylbenzoate 8b were obtained; these steroids inhibited only the 5α-reductase type 2 enzyme. Finasteride and steroids 4, 5, 7b, 8a showed a comparable in vivo pharmacological activity, however the IC50 values of these compounds were higher as compared to that of finasteride. These results indicated also that steroids 4, 5, 7a, and 7b bind to the androgen receptor whereas compounds 6, 8a and 8b failed to do so. The overall data from this study showed that steroids 5 and 7b bind to the AR and decreased of the growth of prostate and seminal vesicles. Moreover, 4 decreased also the growth of seminal vesicles.
- Bratoeff, Eugene,García, Perla,Heuze, Yvonne,Soriano, Juan,Mejía, Adriana,Labastida, Ana María,Valencia, Norma,Cabeza, Marisa
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experimental part
p. 499 - 505
(2010/06/21)
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- An efficient hemisynthesis of 20- and 21-[13C]-labeled cortexolone: A model for the study of skin sensitization to corticosteroids
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A method is described for the synthesis of isotopomers of cortexolone from the commercially available andros-4-ene-3, 17dione. The strategy is based on the use of K13CN for labeling at position 20 and of 13CH 3Mgl, generated in situ, for labeling at position 21. Because of the early introduction of the [13C] labeling, our efforts aimed at reproducible experimental procedures giving high yields with respect to the isotope containing precursors. During the development of this hemisynthesis, we noted that judicious choice of protective groups was essential as this could lead not only to mixtures or unstable intermediates but also influence considerably the output of reactions.
- Claudel, Emilie,Arbez-Gindre, Cecile,Berl, Valerie,Lepoittevin, Jean-Pierre
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scheme or table
p. 3391 - 3398
(2010/02/28)
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- Oxidation of 17α,20β- and 17α,20α-dihydroxypregn-4- en-3-ones, side products of progesterone biotransformation with recombinant microorganisms expressing cytochrome P-45017α
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Progesterone biotransformation with recombinant yeasts Yarrowia lipolytica E129A15 and Saccharomyces cerevisiae GRF18/YEp5117α expressing bovine adrenocortical cytochrome P-45017α yielded 17α-hydroxyprogesterone and two diols, 17α,20β- and 17α,20α-dihydroxypregn-4-en- 3-ones. The oxidation of mixtures of the three steroids with chromic acid resulted in the cleavage of 17-20 bonds in the diols with the formation of androst-4-ene-3,17-dione. The biotransformation of pregn-4-ene-20β-ol-3-one by means of Y. lipolytica E129A15 was accompanied by the following reactions: the primary oxidation of these compounds to progesterone and the subsequent successive reactions of 17α-hydroxylation and 20α- and 20β-reduction. The results widen the possibilities of enzymatic and chemical modifications of steroids.
- Shkumatov,Usova,Radyuk,Kashkan,Kovganko,Juretzek,Mauersberger
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p. 581 - 587
(2007/10/03)
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- Conversion of alpha,beta-unsaturated ketones and alpha,beta-unsaturated esters into alpha-hydroxy ketones and alpha-hydroxy esters using Mn(III) catalyst, phenylsilane and dioxygen
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The present invention provides a novel process for the conversion of α,β-unsaturated ketones. This invention is an improvement over existing processes in that it operates at neutral reaction conditions that prevent the formation of side reactions and that it is a single step, which proceeds with complete selectivity and gives a yield that is approximately 30% higher than the currently used processes. An example of this process is the conversion of 16-dehydroprogesterone into 17 α-hydroxyprogesterone.
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- Conversion of α,β-unsaturated ketones into α-hydroxy ketones using an Mn(III) catalyst, phenylsilane and dioxygen: Acceleration of conjugate hydride reduction by dioxygen
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Treatment of a variety of α,β-unsaturated ketones with Mn(dpm)3 (3 mol%)/PhSiH3 (1.3 equiv.)/isopropyl alcohol/O2, followed by reductive work-up with P(OEt)3 resulted in the formation of α-hydroxy-ketones. (C) 2000 Elsevier Science Ltd.
- Magnus,Payne,Waring,Scott,Lynch
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p. 9725 - 9730
(2007/10/03)
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- Anti-glaucomatous pharmaceutical composition and the process for obtaining them
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The invention relates to the domain of medicinal chemistry. It concerns more particularly that of the preparation of pharmaceutical compositions for ocular use. A subject of the invention is pharmaceutical compositions for ocular use characterized in that they contain at least one selected compound of steroidal structure in combination with or admixed with a pharmaceutically-acceptable, inert carrier or vehicle. The compositions according to the invention are intended to the treatment of glaucoma.
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- HYDROXYLATION OF PROGESTERONE BY CEPHALOSPORIUM APHIDICOLA
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The fungus, Cephalosporium aphidicola, has been shown to hydroxylate progesterone predominantly at the 6β- and 11α-positions.Minor metabolites include tetstosterone acetate, the 20(R)-alcohol and 12β,17α-dihydroxyprogesterone.The sequence involves hydroxylation at 11α and then 6β.The hydroxylations of 11α- and 17α-hydroxyprogesterone and 9β,10α-retroprogesterone have also been examined in the light of these results. - Key words: Cephalosporium aphidicola; progesterone; 9β,10α-retroprogesterone; steroid; microbiological hydroxylation.
- Farooq, Afgan,Hanson, James R.,Iqbal, Zahida
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p. 723 - 726
(2007/10/02)
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- Process for the preparation of steroids bearing 17α-hydroxy-20-oxopregnane side chain
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The invention relates to a novel process for the preparation of 17 α-hydroxy-20-oxopregnane derivatives of the general formula (I), STR1 wherein R1 means a hydroxy or an oxo group; and the dotted lines optionally represent one or more additional valence bond(s), with the proviso that the dotted line in the 4-position and the dotted line in the 5-position cannot each simultaneously be an additional valence bond from steroids having 23,24-dinor-17(20)-dehydrocholan-22-oic -22-oic acid structure. According to the process of the invention a steroid derivative having 23,24-dinorcholan-22-oic acid structure, containing a double bond in 17(20)-position, is transformed to 17α, 20-epoxy-23,24-dinorcholanoic acid, the latter is converted to a reactive acid derivative, which is then reacted with a salt-containing azide ion to yield a 17α, 20-epoxy-23,24-dinorcholanoic acyl azide derivative and the azide obtained is reacted with a mineral or organic acid in an aqueous medium.
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- Tetrapropylammonium perruthenate as a mild and efficient oxidant for sensitive steroidal alcohols
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Tetrapropylammonium perruthenate N-methylmorpholine N-oxide oxidation of steroidal alcohols is described. The reagent combination is mild and gave good yields of the corresponding ketones. Although the oxidation can generate ketones from 3-, 11-, 15-, 17-, and 20-hydroxy steroids, the oxidation of homoallylic alcohols proceeds in low yields. Finally, we observed that the oxidation reagents will convert 17α-hydroxy-2-keto steroids to 17-keto systems in excellent yields.
- Acosta, C. Kirk,Rao, Pemmaraju N.,Kim, Hyun K.
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p. 205 - 208
(2007/10/02)
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- Process for the production of progesterone derivatives
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A process for the production of progresterone derivatives of general formula I STR1 in which symbolizes a single bond or a double bond, X represents a hydrogen atom, a fluorine atom or a methyl group and V means a methylene group, an ethylene group, an ethylidene group or a vinylidene group, is described.
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- THE OXIDATION OF PROGESTERONE UNDER GoAggIII CONDITIONS
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The oxidation of pregn-4-ene-3,20-dione (progesterone) by the GoAggIII system (aqueous hydrogen peroxide, ferric chloride, picolinic acid in pyridine-acetic acid solution) has been investigated.Two tri-keto derivatives were isolated and identified as pregn-4-ene-3,6,20-trione and pregn-4-ene-3,12,20-trione.The third major product isolated was identified as the unstable 5α-formyl-A-nor-pregnane-3,20-dione, which deformylated spontaneously to A-nor-5β-pregnane-3,20-dione.A mechanism for the A-ring contraction is proposed, based upon the participation of a carbon-Fe(V) intermediate.
- Barton, Derek H. R.,Doller, Dario
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p. 984 - 990
(2007/10/02)
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- REDUCTIVE DEHALOGENATION OF 21-IODO DERIVATIVES OF CORTICOSTEROIDS
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In the reactions of 21-iodo derivatives of corticosteroids with hydrogen sulfide and thiol-containing reagents in a medium of dipolar aprotic and amide protogenic solvents at room temperature reductive-deiodination reactions occur with the formation of 21-deoxycorticosteroids in quantitative yield.Reactions of solutions of 21-iodomethyl ketones, heated to 80 deg C, with hydrogen sulfide and thiol-containing reagents give not only reduction products, but also products of nucleophilic substitution at C21 in yields of 20-30percent.
- Mikhal'chuk, A. L.,Pschenichnyi, V. N.
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p. 1479 - 1485
(2007/10/02)
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- 17β-cyano-9α,17α-dihydroxyandrost-4-en-3-one
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The invention is the compound 17β-cyano-9α, 17α-dihydroxyandrost-4-en-3-one (I) which is particularly useful as an intermediate in the production of the 17α-halo silyl ethers (II).
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- Total Synthesis of ( +/-)-Cortisone. Double-Hydroxylation Reaction for Corticoid Synthesis
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Total syntheses of (+/-)-cortisone and (+/-)-adrenosterone have been achieved in 18 steps with the aid of metal-assisted new synthetic sequences in particular, ene reaction and homoenolate chemistry.A novel double-hydroxylation reaction of enol silyl ethers leading to a single step construction of the dihydroxyacetone side chain in corticoids has been developed and applied to the synthesis of cortisone, cortexolone, and 16α-methylcortexolone.
- Horiguchi, Yoshiaki,Nakamura, Eiichi,Kuwajima, Isao
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p. 6257 - 6265
(2007/10/02)
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- Catalytic Hydroxylation of Olefins by Polymer-Bound Osmium Tetroxide
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Osmium tetroxide, linked to insoluble crosslinked polymers and copolymers bearing tertiary amino functions, has been used in the presence of secondary oxidants, such as hydrogen peroxide, tert-butyl hydroperoxide and trimethylamine N-oxide, to accomplish the catalytic hydroxylation of olefins.The polymeric reagent offers the advantages of easy and safe handling, and simple separation from the reaction medium.Generally good yields of vicinal diols have been obtained.
- Cainelli, Gianfranco,Contento, Michele,Manescalchi, Francesco,Plessi, Laura
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- NOVEL OXIDIZING PROPERTIES OF p-METHOXYBENZENETELLURINIC ACID ANHYDRIDE
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The title compound has proved to be a mild oxidizing agent like the corresponding telluroxide or tellurone towards thiol, phosphine, thioamide, thiourea, thionoester, and benzylic alcohol.In addition, it serves as a selective catalyst for the hydration of terminal alkyne.
- Hu, Nan Xing,Aso, Yoshio,Otsubo, Tetsuo,Ogura, Fumio
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p. 6099 - 6102
(2007/10/02)
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- 17β-ethynylsteroids and process for preparing same
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This is disclosed novel intermediates, i.e. 17β-ethynylsteroids, which are useful for the preparation of corticoids such as hydrocortisone and prednisolone, and a process for preparing the same.
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- ANDROSTENEDIONE AND TESTOSTERONE BIOSYNTHESIS BY THE ANDRENAL CORTEX OF THE HORSE
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An homogenate from cortical tissue of mare adrenals was incubated in the presence of tritiated pregnenolone.The (3H) androstenedione and the (3H) testosterone synthesized during the incubation were extracted, purified, and co-crystallized to constant specific activity in the presence of unlabeled carriers.The rate of conversion of pregnenolone to androstenedione and testosterone was of the order of 5 and 0.15 per cent respectively.The high ratio of (3H) androstenedione to (3H) testosterone observed in this study suggests that androstenedione is the main androgen produced by mare adrenals.It is concluded that adrenals could contribute to the production of blood androgens in normal as well as hyperandrogenic mares.
- Silberzahn, Pierre,Rashed, Fakhri,Zwain, Ismail,Leymarie, Pierre
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p. 147 - 152
(2007/10/02)
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- Process for preparing 17α-hydroxy-pregn-4-en-3,20-dione derivatives
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The invention relates to a novel process for the preparation of pregnane derivatives of formula I, STR1 wherein R1 stands for a methyl or an ethyl group, R2 represents a hydrogen atom or a methyl group, and X is a hydrogen atom or a formyl or acetyl group, and the bond indicated by a dotted and a continuous line stands for a single or a double bond between the two neighboring carbon atoms. According to the invention a trifluoroacetate ester of formula II, STR2 wherein R1 and R2 are as defined above, is reacted with formic acid or acetic acid in the presence of a catalytic amount of a mercury salt in a dipolar proton-free or basic solvent. The formyl or acetyl group being in the place of X can be split off in a way known per se. The process provides a novel advantageous method for building up the pregnane side chain characteristic of corticoids.
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- Water-soluble steroid compounds
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Beta-cyclodextrin forms a water-soluble complex or inclusion compound with steroid compounds having a molecular structure smaller than the interior cavity in the doughnut-shaped molecular structure of beta-cyclodextrin. The resulting inclusion compounds can be used for a variety of applications including aqueous topical ophthalmic preparations and topical dermatological ointments.
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- A Ready Conversion of a Pregnane-20-carboxaldehyde to 17α-Hydroxypregnan-20-ones and Pregn-16-en-20-ones
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Reaction of (20S)-5α-chloro-3β-hydroxypregnane-20-carboxaldehyde (9, prepared in two steps from stigmasterol) with oxygen and potassium tert-butoxide, followed by trimethyl phosphite treatment, afforded 5α-chloro-3β,17α-dihydroxypregnan-20-one (6) and 3β,17α-dihydroxypregn-5-en-20-one (7).This mixture was converted into 7 by potassium hydroxide treatment, and into 17α-hydroxypregn-4-ene-3,20-dione (8) by reaction with pyridinium dichromate followed by potassium hydroxide treatment.Bromination-dehydrobromination of 9, in one or two steps, afforded (Z)- and (E)-5α-chloro-3β-hydroxypregn-17(20)-ene-20-carboxaldehyde (11).Reaction of 11 with oxygen and potassium hydroxide gave 5α-chloro-3β-hydroxypregn-16-en-20-one (12), which was converted to 3β-hydroxypregna-15,6-dien-20-one (13) by further potassium hydroxide treatment, and to pregna-4,16-diene-3,20-dione (14) by Oppenauer reaction using excess aluminium isopropoxide.
- Biancini, Bruno,Caciagli, Valerio,Centini, Felice,Eletti-Bianchi, Giancarlo,Re, Luciano
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p. 1829 - 1835
(2007/10/02)
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- An analysis of the metabolites of progesterone produced by isolated Sertoli cells at the onset of gametogenesis
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Sertoli cells isolated from 7 day old rats were maintained in culture and incubated with [14C]-progesterone for 20 h. The cells and media were extracted with ether/chloroform and the extracts chromatographed two-dimensionally on TLC and the radioactive metabolites visualized by autoradiography. Nine of the metabolites (constituting about 88% of total metabolite radioactivity) were identified by relative mobilities of the compounds and their derivatives in TLC and GC systems and by recrystallizations with authentic steroids as the following: 20α-hydroxypregn-4-en-3-one, 3α-hydroxy-5α-pregnan-20-one, 5α-pregnane-3α,20α-diol, 17β-hydroxy-5α-androstan-3-one, 5α-pregnane-3,20-dione, 17-hydroxypregn-4-ene-3,20-dione, testosterone, 5α-androstane-3α,17β-diol and androst-4-ene-3,17-dione. Over 71% of the metabolite radioactivity was due to 20α-hydroxypregn-4-en-3-one, the major metabolite. 5α-reduced pregnanes constituted about 12% and C19 steroids comprised about 2.9% of the radioactivity of the metabolites. Calculation of relative steroidogenic enzyme activities from initial reaction rates suggested the following activities in μunits/mg Sertoli cell protein: 20α-hydroxysteroid oxidoreductase (20α-HSO; 7.71), 5α-reductase (4.77), 3α-HSO (3.57), 17α-hydroxylase (0.93), 17β-HSO (0.34) and C17-C20 lyase (0.34). The relatively high rate of steroidogenic enzyme activities in the Sertoli cells of young rats may indicate that Sertoli cells are less dependent on Leydig cell steroidogenesis than has been assumed. Since nearly all the metabolites of progesterone and testosterone are now identified, it is possible to construct a picture of Sertoli cell steroidogenic activity.
- Wiebe,Tilbe,Buckingham
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p. 561 - 577
(2007/10/02)
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- Novel N-nitroso compounds, compositions containing such compounds, processes for their preparation and methods of treatment therewith, and novel intermediates
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This invention relates to novel N-halogenoalkyl-N-nitroso carbamates and N 4 halogenalkyl-N 4 -nitroso allophanates of steroid compounds, having an anti-tumor activity, and to the preparation thereof. The invention is also concerned with pharmaceutical compositions containing the said compounds, and methods of treatment therewith.
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