Showing papers on "Hydroxysteroid dehydrogenase published in 2010"
TL;DR: It is shown that PFOS and PFOSK are potent inhibitors of rat 3beta-HSD and human 17beta- HSD3 activity, and implies that inhibition of steroidogenic enzyme activity may be a contributing factor to the effects that PFASs exert on androgen secretion in the testis.
50 citations
TL;DR: The ability of soy isoflavones to regulate androgen biosynthesis in Leydig cells is due in part to action on Leydigs cell 3beta-HSD activity, implying that the increasing intake of soy-based food products and their potential effect on blood androgen levels are greatly relevant to public health.
Abstract: The objective of the present study was to investigate the effects of genistein and equol on 3β-hydroxysteroid de- hydrogenase (3β-HSD) and 17β-hydroxysteroid dehydrogenase 3 (17β-HSD3) in human and rat testis microsomes. These enzymes (3β-HSD and 17β-HSD3), along with two others (cytochrome P450 side-chain cleavage enzyme and cytochrome P450 17α-hydroxylase/17-20 lyase), catalyze the reactions that convert the steroid cholesterol into the sex hormone testosterone. Genistein inhibited 3β-HSD activity (0.2 µmol L -1 pregnenolone) with half-maximal in- hibition or a half-maximal inhibitory concentration (IC50) of 87 ± 15 (human) and 636 ± 155 nmol L -1 (rat). Genistein's mode of action on 3β-HSD activity was competitive for the substrate pregnenolonrge and noncompetitive for the co- factor NAD + . There was no difference in genistein's potency of 3β-HSD inhibition between intact rat Leydig cells and testis microsomes. In contrast to its potent inhibition of 3β-HSD, genistein had lesser effects on human and rat 17β-HSD3 (0.1 µmol L -1 androstenedione), with an IC50 ≥ 100 µmol L -1 . On the other hand, equol only inhibited hu- man 3β-HSD by 42%, and had no effect on 3β-HSD and 17β-HSD3 in rat tissues. These observations imply that the ability of soy isoflavones to regulate androgen biosynthesis in Leydig cells is due in part to action on Leydig cell 3 β- HSD activity. Given the increasing intake of soy-based food products and their potential effect on blood androgen levels, these findings are greatly relevant to public health.
47 citations
TL;DR: The synthesis and SAR studies of 3- and 4-substituted 7-hydroxycoumarins as novel 17beta-HSD3 inhibitors are discussed and the most potent compounds from this series exhibited low nanomolar inhibitory activity with acceptable selectivity versus other 17 beta- HSD isoenzymes and nuclear receptors.
47 citations
TL;DR: In vitro testing revealed high inhibitory potencies toward human placental 17β-HSD1, and the most promising compounds, 14 and 15, showed IC(50) values in the low nanomolar range in the cell-free and cellular assays, which make these inhibitors interesting candidates for further preclinical evaluation.
Abstract: Estradiol (E2), the most important estrogen in humans, is involved in the initiation and progression of estrogen-dependent diseases such as breast cancer and endometriosis. Its local production in ...
46 citations
TL;DR: Structures and kinetic properties of the wild-type and mutant enzymes indicate that Leu308 is a selectivity determinant for inhibitor binding, and 5-substituted 3-chlorosalicylic acid derivatives were synthesized, of which the most potent compound was 24-fold more selective for AKR1C1 relative to the structurally similar 3α-hydroxysteroid dehydrogenase.
20 citations
TL;DR: Non-steroidal compounds that inhibit 17beta-hydroxysteroid dehydrogenase isoform 3, an enzyme catalyzing the final step in testosterone biosynthesis in Leydig cells, are under development for male contraceptive or treatment of androgen dependent diseases including prostate cancer.
20 citations
TL;DR: It is demonstrated that under conditions that avoided changes in food intake seen in gene knockout or higher-dose pharmacologic models, the efficacy of 11beta-HSD1 inhibition to up-regulate hepatic fat oxidation gene expression functionally translates into enhanced hepatic lipid oxidation in vivo.
Abstract: This study aimed to explore in a model of diet-induced steatosis the impact of pharmacologic 11 β -hydroxysteroid dehydrogenase type 1 (11 β -HSD1) inhibition, under conditions of unchanged ingestive behavior, on liver fat oxidation. Male Sprague-Dawley rats were fed an obesogenic diet and were continuously treated or not with an 11 β -HSD1 inhibitor (Compound A, 3 mg/[kg d]; Merck Research Laboratories, Rahway, NJ), after which liver expression of oxidative genes and in vivo hepatic fat oxidation were quantified. Treatment with Compound A reduced liver triglyceride concentration (−28%), increased hepatic expression of several genes coding for enzymes of mitochondrial and peroxisomal β -oxidation, and concomitantly enhanced in vivo liver fat oxidation (+38%). The study demonstrates, under conditions that avoided changes in food intake seen in gene knockout or higher-dose pharmacologic models, the efficacy of 11 β -HSD1 inhibition to up-regulate hepatic fat oxidation gene expression, which functionally translates into enhanced hepatic lipid oxidation in vivo.
18 citations
TL;DR: Sequencing of genomes from sea urchin, a basal deuterostome, and amphioxus, and the availability of substantial sequence for acorn worm and elephant shark provide a more complete dataset for analysis of the origins of 11beta-HSD2 and 17beta- HSD2, which are paralogs of human testosterone and estradiol.
18 citations
TL;DR: The role of a single amino acid in the evolution of these functionally diverse enzymes is demonstrated for the first time and new physiological functions for 17beta-HSD1 in retinoid metabolism are suggested.
17 citations
TL;DR: An AKR-dependent whole-cell biotransformation process that can be used for production of human AKR metabolites on a large scale is established.
14 citations
TL;DR: A series of novel adamantyl ethanone compounds was identified as potent inhibitors of human 11β‐HSD1 and docking studies of the potent compounds revealed how these molecules may interact with the enzyme and cofactor.
Abstract: 11Beta-hydroxysteroid dehydrogenases (11beta-HSDs) are key enzymes regulating the pre-receptor metabolism of glucocorticoid hormones. The modulation of 11beta-HSD type 1 activity with selective inhibitors has beneficial effects on various conditions including insulin resistance, dyslipidemia and obesity. Inhibition of tissue-specific glucocorticoid action by regulating 11beta-HSD1 constitutes a promising treatment for metabolic and cardiovascular diseases. A series of novel adamantyl ethanone compounds was identified as potent inhibitors of human 11beta-HSD1. The most active compounds identified (52, 62, 72, 92, 103 and 104) display potent inhibition of 11beta-HSD1 with IC(50) values in the 50-70 nM range. Compound 72 also proved to be metabolically stable when incubated with human liver microsomes. Furthermore, compound 72 showed very weak inhibitory activity for human cytochrome P450 enzymes and is therefore a candidate for in vivo studies. Comparison of the publicly available X-ray crystal structures of human 11beta-HSD1 led to docking studies of the potent compounds, revealing how these molecules may interact with the enzyme and cofactor.
TL;DR: The present data are in agreement with the bifunctional activities of 17β-HSD12 suggesting that the VLCFA elongation activity, having its origin in the yeast, is most probably responsible for embryonic lethality in HSD17B12–/–, whereas the more recently acquired 17 β- HSD12 activity is responsible for reduced sex steroid levels in H SD17B 12+/–.
Abstract: 17β-Hydroxysteroid dehydrogenases (17β-HSDs) are enzymes issued from convergent evolution of activity from various ancestral genes having different functions. Type 12 17β-HSD (17β-HSD12) was described as a bifunctional enzyme, involved in the biosynthesis of estradiol (E2) and the elongation of very long chain fatty acid (VLCFA). It catalyzes selectively the transformation of estrone (E1) into estradiol (E2) in human and primates, whereas in the mouse and Caenorhabditis elegans the enzyme catalyzes the 17β-reduction of both androgens and estrogens. It is also able to catalyze the reduction of 3-keto-acylCoA into 3-hydroxy-acylCoA in the elongation cycle of VLCFA biosynthesis. To further understand the physiological role of 17β-HSD12, we performed targeted disruption of the Hsd17b12 gene by substituting exons 8 and 9 that contain the active site with a neomycin cassette. The data indicate that heterozygous (HSD17B12+/-) mice are viable with reduced levels of sex steroids, whereas homozygous (HSD17B12-/-) mice show embryonic lethality. The present data are in agreement with the bifunctional activities of 17β-HSD12 suggesting that the VLCFA elongation activity, having its origin in the yeast, is most probably responsible for embryonic lethality in HSD17B12-/-, whereas the more recently acquired 17β-HSD12 activity is responsible for reduced sex steroid levels in HSD17B12+/-.
TL;DR: In this paper, the synthesis and SAR studies of 3 and 4-substituted 7-hydroxycoumarins as novel 17β-HSD3 inhibitors are discussed.
Abstract: The synthesis and SAR studies of 3- and 4-substituted 7-hydroxycoumarins as novel 17β-HSD3 inhibitors are discussed. The most potent compounds from this series exhibited low nanomolar inhibitory activity with acceptable selectivity versus other 17β-HSD isoenzymes and nuclear receptors.
TL;DR: The new bacterial expression system for recombinant 17β-HSD1 is useful for the easy purification of large amounts and will facilitate the functional study of this enzyme.
Abstract: Human 17β-hydroxysteroid dehydrogenase type 1 (17β-HSD1) catalyzes the reaction of estrone with NADPH to form estradiol and NADP+, thereby regulating the biological activity of sex steroid hormones in a variety of tissues. Here, we present an efficient method for expressing and purifying human 17β-HSD1 from Escherichia coli. The expression vector pET28a/17β-HSD1 was constructed and transformed into Escherichia coli BL21(DE3) cells. We found that the active enzyme can be obtained by inducing 17β-HSD1 expression at 0.25 mM IPTG, 13°C for overnight. The protein is purified by single step Ni–NTA affinity chromatography and yields 2.8 mg/L of culture. The kinetic study shows V/Et of (1.21 ± 0.05) × 10−2/s and Kestradiol of 0.8 μM in the oxidation of estradiol with NADP+ as cofactor at pH 9.3. The new bacterial expression system for recombinant 17β-HSD1 is useful for the easy purification of large amounts and will facilitate the functional study of this enzyme.
TL;DR: It is found that melatonin decreases the growth of C6 glioma cells and reduces the sulfatase and 17beta-hydroxysteroid dehydrogenase activity.
Abstract: Melatonin interacts with estradiol at the estrogen receptor level in different kinds of neoplasias and also regulates the expression and the activity of some enzymes involved in the biosynthesis of estrogens in peripheral tissues. Glioma cells express estrogen receptors and have the ability to synthesize estrogens locally. Since melatonin inhibits the growth of C6 cells, and this indoleamine has been demonstrated to be capable of decreasing aromatase expression and activity in these cells, the aim of the present study was to analyze whether the regulation of the sulfatase, the enzyme that catalyzes the rate-limiting step in the conversion of estrogen sulfates to estrogens, and 17beta-hydroxysteroid dehydrogenase, the enzyme which converts the relatively inactive estrone to the most potent 17beta-estradiol, could be involved in the inhibition of glioma cell growth by melatonin. We found that melatonin decreases the growth of C6 glioma cells and reduces the sulfatase and 17beta-hydroxysteroid dehydrogenase activity. Finally, we demonstrated that melatonin downregulates sulfatase and 17beta-hydroxysteroid dehydrogenase mRNA steady state levels in these glioma cells. By analogy to the implications of these enzymes in other forms of estrogen-sensitive tumors, it is conceivable that their modulation by melatonin may play a role in the growth of glioblastomas.
TL;DR: A comparison of the structures and kinetic properties of human 20alpha-hydroxysteroid dehydrogenase (AKR1C1) and its mutant enzymes (Leu308Val and Leu308Ala) indicates that Leu 308 is a selectivity determinant for substrate binding.
TL;DR: Mouse 3( 17)alpha-hydroxysteroid dehydrogenase (AKR1C21) is the only aldo-keto reductase that catalyzes the stereospecific reduction of 3- and 17-ketosteroids to the corresponding 3(17)Alpha-Hydroxysteroids, and mutations of residues involved in forming the safety belt for binding of the coenzyme showed similar alterations in kinetic constants compared with the wild type.
Abstract: Mouse 3(17)alpha-hydroxysteroid dehydrogenase (AKR1C21) is the only aldo-keto reductase that catalyzes the stereospecific reduction of 3- and 17-ketosteroids to the corresponding 3(17)alpha-hydroxysteroids. The Y224D mutation of AKR1C21 reduced the K(m) value for NADP(H) by up to 80-fold and completely reversed the 17alpha stereospecificity of the enzyme. The crystal structure of the Y224D mutant at 2.3 A resolution revealed that the mutation resulted in a change in the conformation of the flexible loop B, including the V-shaped groove, which is a unique feature of the active-site architecture of wild-type AKR1C21 and is formed by the side chains of Tyr224 and Trp227. Furthermore, mutations (Y224F and Q222N) of residues involved in forming the safety belt for binding of the coenzyme showed similar alterations in kinetic constants for 3alpha-hydroxy/3-ketosteroids and 17-hydroxy/ketosteroids compared with the wild type.
TL;DR: Novel estradiol-imidazole C-nucleoside hybrid compounds 4a and 4b, which have C4-linked C o - and C 2 -imidzole ribonucleosides as adenosine mimics and amide bond linkers, were designed and synthesized based on EM-1745, an inhibitor of type 1 17β-hydroxysteroid dehydrogenase.
Abstract: Novel estradiol-imidazole C-nucleoside hybrid compounds 4a and 4b, which have C4-linked C o - and C 2 -imidazole ribonucleosides as adenosine mimics and amide bond linkers, were designed and synthesized based on EM-1745, an inhibitor of type 1 17β-hydroxysteroid dehydrogenase (17β-HSD1). Compounds 4a and 4b were also tested as enzyme inhibitors.
Patent•
13 Oct 2010
TL;DR: In this paper, a 12 alpha -hydroxysteroid dehydrogenase (12alpha -HSDH) mutant catalyzing at least stereospecific enzymatic oxidation of a 12-hydroxylsteroid to the corresponding 12-ketosteroid, is new, where the mutant exhibits an increased specific activity (U/mg) in the presence of NADP +>cofactor in comparison to non-mutated enzyme.
Abstract: 12alpha -Hydroxysteroid dehydrogenase (12alpha -HSDH) mutant catalyzing at least stereospecific enzymatic oxidation of a 12-hydroxysteroid to the corresponding 12-ketosteroid, is new, where the mutant exhibits an increased specific activity (U/mg) in the presence of NADP +>cofactor in comparison to non-mutated enzyme. Independent claims are included for: (1) a nucleic acid sequence encoding the 12alpha -HSDH mutant; (2) an expression cassette comprising the nucleic acid sequence under the genetic control of at least one regulatory nucleic acid sequence; (3) a vector comprising at least one expression cassette; (4) a recombinant microorganism which carries at least one nucleic acid sequence or at least one expression cassette or is transformed with at least one vector; (5) enzymatic synthesis of 12alpha -hydroxysteroids, comprising reacting the corresponding 12-ketosteroid in the presence of 12alpha -HSDH mutant, and optionally isolating at least one reduction product formed from the reaction mixture; (6) enzymatic oxidation of 12alpha -hydroxysteroids, comprising reacting the hydroxysteroid in the presence of 12alpha -hydroxysteroid dehydrogenase, and optionally isolating an oxidation product formed from the reaction mixture; and (7) preparing ursodeoxycholic acid of formula (I), comprising oxidizing a cholic acid of formula (II) in the presence of 12alpha -hydroxysteroid dehydrogenase mutant for the corresponding 12-ketochenodeoxycholic acid of formula (III), and subsequently reacting (III) by deoxygenation to chenodeoxycholic acid of formula (IV), chemically oxidizing (IV) in position 7 to 7-keto-lithocholic acid of formula (V), reducing (V) and optionally further purifying the reaction product. R : alkyl, NR1R2, H, alkali metal ion or N(R3) 4+>; R3 : H or alkyl; and R1a : H or acyl. [Image] [Image] ACTIVITY : Hepatotropic; Litholytic. MECHANISM OF ACTION : None given.
Patent•
10 Dec 2010
TL;DR: In this article, a method for enzymatic oxidation of 12 alpha -hydroxy steroid, comprising converting the hydroxysteroid in the presence of 12alpha-hydroxysteroid dehydrogenase and cofactor regenerating system as above per se and optionally isolating at least one of the formed oxidation product from the reaction mixture, was presented.
Abstract: Method for cofactor regeneration, comprises redox reaction in which a substrate is converted to corresponding oxidized or reduced product under the use of at least one redox-enabled cofactor and regenerating the used cofactor under the use of an enzymatic cofactor regenerating system comprising (a) a NAD(P)H dehydrogenase with cholic acid tolerance and (b) a redox-mediator, which transfers electrons to molecular oxygen. Independent claims are included for: (1) a method for enzymatic oxidation of 12alpha -hydroxy steroid, comprising converting the hydroxysteroid in the presence of 12alpha -hydroxysteroid dehydrogenase and cofactor regenerating system as above per se and optionally isolating at least one of the formed oxidation product from the reaction mixture; and (2) preparing ursodeoxycholic acid of formula (I), comprising (a1) enzymatically oxidizing cholic acid compounds of formula (II) in the presence of 12alpha -hydroxysteroid dehydrogenase and the cofactor regenerating system as above per se to corresponding 12-keto-chenodeoxycholic acid compounds of formula (III), (b1) deoxygenating (III) to chenodeoxycholic acid of formula (IV), (c1) chemically oxidizing (IV) in position 7 to give 7-keto-lithocholic acid of formula (V), (d1) reducing (V) and (e1) optionally further purifying the reaction product. R : alkyl, NR11R2, H, alkali metal ion or N(R3) 4+>; R3 : H or alkyl; R1 : H or acyl; and R11, R2 : not defined. [Image] [Image] [Image].
Patent•
22 Apr 2010
TL;DR: A treatment using a hydroxysteroid dehydrogenase reductase inhibitor combined with a mineralocorticoid receptor antagonist is provided in this paper, which overcomes the problem that manipulating Cortisol levels by a single compound therapy induces other pathologies or side effects.
Abstract: A treatment using a hydroxysteroid dehydrogenase reductase inhibitor combined with a mineralocorticoid receptor antagonist is provided which overcomes the problem that manipulating Cortisol levels by a single compound therapy induces other pathologies or side effects, while the original condition that required treatment is ameliorated.
01 Jan 2010
TL;DR: High level of substrates and low level of enzymes indicate the inhibition of steroidogenesis in treated mice and may be due to the presence of sterols.
Abstract: The petroleum ether extract of Celsia coromandeliane Vahl (PECC) arrested the normal estrus cycle of adult female mouse and significantly decreased the weight of ovaries and uterus. The cholesterol and ascorbic acid contents in ovaries were significantly increased in treated mice. Two key enzymes, 5 -3– hydroxysteroid dehydrogenase ( 5 -3-HSD) and glucose-6-phosphate dehydrogenase (G-6-PDH), were decreased significantly in PECC treated mice after 17 days of treatment. High level of substrates and low level of enzymes indicate the inhibition of steroidogenesis in treated mice and may be due to the presence of sterols. Keywods : Celsia coromandeliane , 5 -3–hydroxysteroid dehydrogenase(HSD), glucose-6-phosphate dehydrogenase, mouse ovary.
Patent•
03 Feb 2010Patent•
23 Jul 2010
TL;DR: In this article, a compound of the general formula (I) is defined as a compound useful in modulating the activity of 11β-hydroxysteroid dehydrogenase type 1 (11βHSD1) and is useful for treating pathologies in which such modulation is beneficial, as in the case of metabolic syndrome or of noninsulin dependent type 2 diabetes.
Abstract: The invention relates to a compound of the general formula (I),
as defined herein which is useful in modulating the activity of 11β-hydroxysteroid dehydrogenase type 1 (11βHSD1) and are useful for treating pathologies in which such modulation is beneficial, as in the case of metabolic syndrome or of noninsulin-dependent type 2 diabetes. The invention also relates to pharmaceutical preparations containing such a compound, processes for preparing and intermediates useful in the preparation of a such a compound.
Patent•
13 Sep 2010
TL;DR: Use of a compound represented by formula (I), a salt of the compound, or a solvate of the compounds or the salt for inhibiting type-3 17β-hydroxysteroid dehydrogenase; and others as mentioned in this paper.
Abstract: Use of a compound represented by formula (I), a salt of the compound, or a solvate of the compound or the salt for inhibiting type-3 17β-hydroxysteroid dehydrogenase; and others.