Showing papers on "Hydroxysteroid dehydrogenase published in 2009"
TL;DR: The distribution of the zearalenone reducing activity was investigated in liver fractions obtained by differential centrifugation of liver homogenate from adult female Sprague Dawley rats and it was found that at least two multiple forms occur of the enzyme with different subcellular locations and pH-optima.
Abstract: The distribution of the zearalenone reducing activity was investigated in liver fractions obtained by differential centrifugation of liver homogenate from adult female Sprague Dawley rats. The zearalenone reducing enzyme was identified as 3 alpha-hydroxysteroid dehydrogenase. At least two multiple forms occur of the enzyme with different subcellular locations and pH-optima. The activity was localized in the microsomes with NADH as coenzyme and in both microsomes and cytosol with NADPH.
89 citations
TL;DR: The fluorine compound 23 exhibits an IC(50) of 8 nM and is the most potent nonsteroidal inhibitor described so far and shows a high selectivity (17beta-HSD2, ERalpha) and excellent pharmacokinetic properties after peroral application to rats.
Abstract: 17β-Hydroxysteroid dehydrogenase type 1 (17β-HSD1) is responsible for the catalytic reduction of weakly active E1 to highly potent E2. E2 stimulates the proliferation of hormone-dependent diseases ...
79 citations
TL;DR: The data suggest that 17β‐HSD8 is primarily involved in mitochondrial FAS, and these two proteins display a stable physical interaction and form an active heterotetramer, which is likely to form the long‐sought HsKAR.
Abstract: Mitochondrial fatty acid synthesis (FAS) generates the octanoyl-group that is required for the synthesis of lipoic acid and is linked to mitochondrial RNA metabolism. All of the human enzymes involved in mitochondrial FAS have been characterized except for β-ketoacyl thioester reductase (HsKAR), which catalyzes the second step in the pathway. We report here the unexpected finding that a heterotetramer composed of human 17β-hydroxysteroid dehydrogenase type 8 (Hs17β-HSD8) and human carbonyl reductase type 4 (HsCBR4) forms the long-sought HsKAR. Both proteins share sequence similarities to the yeast 3-oxoacyl-(acyl carrier protein) reductase (Oar1p) and the bacterial FabG, although HsKAR is NADH dependent, whereas FabG and Oar1p are NADPH dependent. Hs17β-HSD8 and HsCBR4 show a strong genetic interaction in vivo in yeast, where, only if they are expressed together, they rescue the respiratory deficiency and restore the lipoic acid content of oar1Δ cells. Moreover, these two proteins display a stable physica...
51 citations
TL;DR: Compared (+)- and (-)-gossypols in the inhibition of 3beta-hydroxysteroid dehydrogenase and 17beta-HSD isoform 3 in human and rat testes, gossypol enantiomers competitively inhibited both 3beta -HSD and 17 beta- HSD3 by competing for the cofactor binding sites of these enzymes.
49 citations
TL;DR: KLF15 is potentially a novel link between the regulation of testosterone production and fat stores by insulin in humans and its potential role in the pathogenesis of hyperandrogenism is presented.
Abstract: Context: Kruppel-like factor 15 (KLF15) is a newly discovered transcription factor that plays an important role in glucose homeostasis and lipid accumulation in cells. We present evidence for KLF15 as a transcriptional regulator of the human 17β-hydroxysteroid dehydrogenase type 5 gene (HSD17B5) and its potential role in the pathogenesis of hyperandrogenism. Objective: The aim was to investigate the molecular mechanism of HSD17B5 regulation. Methods: Diverse molecular biology techniques were used. Design and Results: We identified a KLF15 binding site in the HSD17B5 promoter by using luciferase promoter constructs, EMSA, and chromatin immunoprecipitation assays. Overexpression of KLF15 increased HSD17B5 promoter activity and testosterone formation at least 3-fold in cultured H295R cells. Insulin increased KLF15 mRNA expression according to real-time RT-PCR and increased HSD17B5 promoter activity according to luciferase assays. KLF15 overexpression in combination with insulin, glucocorticoid, and cAMP stim...
48 citations
TL;DR: A novel set of potent inhibitors of steroidogenic human 17beta-hydroxysteroid dehydrogenase type 1 (17beta-HSD 1) is developed and the inhibitory potency of compounds is comparable or better to that of previously described inhibitors.
46 citations
TL;DR: Estrone C15 derivative compound 21 can be regarded as a promising lead compound for further development as a 17beta-HSD1 inhibitor and did not show estrogen receptor mediated effects in vitro or in vivo.
40 citations
TL;DR: Compound 26, a 3-deoxyestradiol derivative with a dimethylated spiro-delta-lactone at position 17, possesses the most potent inhibitory activity for 17beta-HSD5 (IC(50)=2.9 nM), and showed no binding affinity for estrogen, androgen, progestin and glucocorticoid receptors (ER, AR, PR and GR).
39 citations
TL;DR: A library of fused (di)cycloalkeno thieno[2,3-d]pyrimidin-4(3H)-one based compounds was synthesized and the majority of these compounds exhibited excellent selectivity over the oxidative isoform 17beta-HSD2 and lacked estrogenic effects in an estrogen receptor (ER) binding assay.
Abstract: Many breast tumors are hormone-dependent, and estrogens, especially estradiol (E2), have a pivotal role in their growth and development. 17β-Hydroxysteroid dehydrogenase type 1 (17β-HSD1) is a key enzyme in the biosynthesis of female sex steroids, catalyzing the NADPH-dependent reduction of estrone into biologically active estradiol. In this study, a library of fused (di)cycloalkeno thieno[2,3-d]pyrimidin-4(3H)-one based compounds was synthesized, and the biological activities against 17β-HSD1 in a cell-free and in a cell-based assay were evaluated. Several thieno[2,3-d]pyrimidin-4(3H)-one based compounds, at 0.1 and 1 μM test concentrations, were found to be potent 17β-HSD1 inhibitors. For example, 4-(3-hydroxyphenylthio)-1,2,7,8,9,10,11,13-octahydro-13-oxo-[1]benzothieno[2′,3′:4,5]-pyrimido[1,2-a]azepine-3-carboxaldehyde (7f) is one of the most potent nonsteroidal 17β-HSD1 inhibitors reported to date with 94% inhibition of the recombinant enzyme at 0.1 μM test concentration. Importantly, the majority of...
39 citations
TL;DR: The findings strongly support suitability of non-steroidal 17betaHSD1-inhibitors for the treatment of estrogen dependent diseases.
38 citations
TL;DR: The data indicate that there is a low conversion of E1 into E2 in preadipocytes; however this activity is increased approximately 5-fold (p<0.0001) in differentiated adipocytes, which is consistent with the increase in protein expression levels of 17beta-HSD12.
TL;DR: The first design, synthesis, and evaluation of human 20alpha-hydroxysteroid dehydrogenase (AKR1C1) inhibitors based on the recently published crystal structure of its ternary complex with inhibitor are reported.
Abstract: The first design, synthesis, and evaluation of human 20α-hydroxysteroid dehydrogenase (AKR1C1) inhibitors based on the recently published crystal structure of its ternary complex with inhibitor are reported. While the enzyme−inhibitor interactions observed in the crystal structure remain conserved with the newly designed inhibitors, the additional phenyl group of the most potent compound, 3-bromo-5-phenylsalicylic acid, targets a nonconserved hydrophobic pocket in the active site of AKR1C1 resulting in 21-fold improved potency (Ki = 4 nM) over the structurally similar 3α-hydroxysteroid dehydrogenase isoform (AKR1C2). The compound is hydrogen bonded to Tyr55, His117, and His222, and the phenyl ring forms additional van der Waals interactions with residues Leu308, Phe311, and the nonconserved Leu54 (Val in AKR1C2). Additionally, the metabolism of progesterone in AKR1C1-overexpressed cells was potently inhibited by 3-bromo-5-phenylsalicylic acid, which was effective from 10 nM with an IC50 value equal to 460 nM.
TL;DR: Using a structure- and ligand-based approach, a pharmacophore model was proposed and a new class of non-steroidal inhibitors of 17beta-HSD1 was designed, and the potency of this class of inhibitors was improved by substitution of the 1-position of the naphthalene ring by a phenyl group.
TL;DR: First selective and efficient inhibitors of 17beta-HSD7 are now available for additional mechanistic and therapeutic studies.
Abstract: 17β-Hydroxysteroid dehydrogenase type 7 (17β-HSD7) catalyzes the reduction of estrone (E1) into estradiol (E2) and of dihydrotestosterone (DHT) into 5α-androstane-3β,17β-diol (3β-diol), therefore modulating the level of mitogenic estrogens and androgens in humans. By classical and parallel chemistry, we generated several 4-methyl-4-aza-5α-androstane derivatives differing in their C-17 substituent: 17β-formamide, 17β-benzamide, and 17β-tertiary amine. Best candidates in each category had demonstrated good inhibitory potency toward the conversion of E1 into E2 (IC50 = 189−451 nM) and also toward the conversion of DHT into 3β-diol (69−91% at 3 μM). Inhibition assays with 17β-HSD1, 17β-HSD5, 5α-reductase (5α-R) 1 and 5α-R2 revealed that 17β-HSD7 inhibitors with a 4-methyl-4-aza nucleus were also able to inhibit 5α-Rs but not the other enzymes tested. Two 4-aza-5α-androstane inhibitors were, however, selective and still showed good inhibition of 17β-HSD7. First selective and efficient inhibitors of 17β-HSD7 ar...
TL;DR: Real-time RT-PCR and Western blotting showed an induction of 20beta-HSD expression during human chorionic gonadotropin (hCG)-induced oocyte maturation, in vitro and in vivo, and specific inhibitors of carbonyl reductase inhibited not only recombinant protein catalytic activity but also hCG-induced oocytes maturation in a dose-dependent manner as evidenced by blocking of germinal vesicle break down.
TL;DR: For the reductive HSDs human 17betaHSD type 1 and rat AKR1C9, one can attenuate or reverse the directional preference in intact cells by site-directed mutagenesis in the cofactor-binding domain or by glucose deprivation, but the magnitude of such changes vary with the different enzymes.
TL;DR: Cl cloning and characterization of a novel human SDR gene SCDR10B which encodes a protein with similarity to 11beta-hydroxysteroid dehydrogenase 1 which can catalyze the dehydrogenation of cortisol in the presence of NADP(+), and therefore it is a hydroxysteroid dehydrationase.
Abstract: Hydroxysteroid dehydrogenase belongs to the subfamily of short-chain dehydrogenases/reductases (SDR), and 11-beta-hydroxysteroid dehydrogenase catalyzes the interconversion of inactive glucocorticoids (cortisone in human, dehydrocorticosterone in rodents) and active glucocorticoids (cortisol in human, corticosterone in rodents). We report here the cloning and characterization of a novel human SDR gene SCDR10B which encodes a protein with similarity to 11beta-hydroxysteroid dehydrogenase 1. SCDR10B was isolated from a human brain cDNA library, and was mapped to chromosome 19p13.3 by browsing the UCSC genomic database. It contains an ORF with a length of 858 bp, encoding a protein with a transmembrane helix and SDR domain. Its molecular mass and isoelectric point are predicted to be 30.8 kDa and 10.3 kDa, respectively. SCDR10B protein is highly conserved in mammals and fish. Phylogenetic tree analysis indicated that SCDR10B stands for a new subgroup in the 11beta-hydroxysteroid dehydrogenase family. Northern blot analysis showed that SCDR10B was highly expressed in brain, and a strong expression signal was detected in hippocampal neurons by immunohistochemical analysis. RT-PCR and immunohistochemical analysis showed that SCDR10B was up-regulated in lung-cancer cell lines and human lung cancer. SCDR10B can catalyze the dehydrogenation of cortisol in the presence of NADP(+), and therefore it is a hydroxysteroid dehydrogenase.
TL;DR: The synthesis and SAR of a series of arylsulfonylpiperazine inhibitors of 11beta-HSD1 led to potent, selective, and orally bioavailable inhibitors demonstrating efficacy in a cynomolgus monkey ex vivo enzyme inhibition model.
TL;DR: It is shown that HSD17B8 expression is up-regulated in response to E2 in the estrogen receptor alpha (ERalpha) positive MCF-7 cells, and co-immunoprecipitation studies revealed tethering of ERalphatoC/EBPbeta in response-to-E2 in cells expressing ERalpha.
Abstract: Hydroxysteroid (17-beta) dehydrogenase (HSD17B) are the enzymes responsible for the reversible interconversion of 17-hydroxy and 17-keto steroids. The human and mouse type 8 17beta-HSD (HSD17B8) selectively catalyze the conversion of estradiol (E2) to estrone (E1). We previously described thatHSD17B8 is transcriptionally regulated by C/EBPbeta, and that C/EBPbeta is bound to CCAAT boxes located at -5 and -46 of the transcription start site in basal conditions in HepG2 cells. Furthermore, ectopic expression of C/EBPbeta transactivated the HSD17B8 promoter activity. Here, we show that HSD17B8 expression is up-regulated in response to E2 in the estrogen receptor alpha (ERalpha) positive MCF-7 cells. Results showed that this induction is mediated by ERalpha because i) E2 did not induce HSD17B8 expression in ERalphanegative HepG2 cells, ii) ectopic expression of ERalpha restored E2-induced HSD17B8 expression, and iii) this induction was blocked by the anti-ER ICI 182,780. Additional experiments showed that no estrogen response element was necessary for this regulation. However, the CCAAT boxes located at the HSD17B8 proximal promoter were required for E2-induced transcription. Furthermore, co-immunoprecipitation studies revealed tethering of ERalphatoC/EBPbeta in response to E2 in cells expressing ERalpha. Additionally, chromatin immunoprecipitation assays demonstrated that, in response to E2, ERalpha is recruited to the CCAAT boxes in which C/EBPbeta is already bound. Taken together, our results reveal that ERalpha is involved in the transcriptional regulation of HSD17B8 gene in response to E2 through its interaction with C/EBPbeta.
TL;DR: In this paper, a series of adamantyl carboxamides were used as selective inhibitors of human 11β-hydroxysteroid dehydrogenases (11β-HSDs) in HEK-293 cells transfected with the HSD11B1 gene.
TL;DR: Identification of protein interaction partners by yeast two-hybrid system suggests that despite the potential lack of enzymatic activity HSDL1 might retain regulatory functions in the cell.
TL;DR: Results obtained in this study clearly indicate that SREBP-1 represents one of the transcriptional regulators of human 17beta-HSD12.
TL;DR: A series of pyrimidine, phthalimido and athranilic acid derivatives are synthesized and examined their inhibitory properties towards AKR1C1, a member of the aldo-keto reductase superfamily and potential agents for treatment of endometrial cancer and endometriosis.
TL;DR: In vitro biological evaluation of these steroid derivatives revealed that a spacer of 13 methylenes, between the 16β-position of E2 and the adenosine mimic bearing a carboxylic acid group, gave the best inhibition of 17β-HSD1.
Abstract: A series of estradiol (E2) derivatives were designed to interact with both the substrate- and the cofactor-binding sites of 17β-hydroxysteroid dehydrogenase type 1 (17β-HSD1). These analogues of po...
TL;DR: Results strongly support an additional 17beta-HSD1 binding site for phytoestrogens which is neither the substrate nor the co-factor binding site, and may open new opportunities for the design of inhibitors, not only for 17 beta- HSD1, but also for other family members of the short chain dehydrogenases.
TL;DR: In this article, a bisubstrate inhibitor of 17β-HSD1, the estradiol/adenosine hybrid EM-1745, is reviewed and strategies for future designs of inhibitors are proposed.
Abstract: Type 1 17β-hydroxysteroid dehydrogenase (17β-HSD1) is a key steroidogenic enzyme that catalyses the reduction of steroid estrone into the most potent endogenous estrogen estradiol using the cofactor NAD(P)H. Bisubstrate inhibition is a good way to enhance the potency of inhibitors of cofactor-assisted enzymes. The design of a bisubstrate inhibitor of 17β-HSD1, the estradiol/adenosine hybrid EM-1745, is reviewed and strategies for future designs of inhibitors are proposed.
TL;DR: Cyproteron‐Acetat‐induzierte Veränderungen im Verhalten der Hydroxysteroid‐Dehydrogenase in den Leydig‐Zellen bei der Ratte während der perinatalen Entwicklung.
Abstract: The influence of cyproterone acetate (CA) upon the behaviour of hydroxysteroid dehydrogenases (HSDH) in the Wistar rat Leydig cells was investigated during the perinatal phase with the help of enzymhistochemical cum morphometrical techniques. The pregnant rats as well as their offsprings were injected with CA (dosage: 35 mg/kg body wt) sc, daily from 14 fetal day upto 31 postnatal day (p.n.d.). The animals were killed on 5, 20, and 32 p.n.d.; the enzymhistochemical reactions for 3-beta-HSDH, 11-beta-HSDH, 17-HSDH and 3-alpha-HSDH were performed in the cryostat sections of the testis, and the morphometric evaluation of HSDH positive Leydig cells was carried out. On 5 p.n.d. the activity of 17-beta-HSDH was slightly impaired in the intertubular Leydig cells of the CA treated animals. On 20 p.n.d., CA prevented nearly completely the HSDH activity in the newsly built peritubular Leydig cells; the activities of 3-beta-HSDH, 17-beta-HSDH, and 3-alpha-HSDH resided mainly in the intertubular Leydig cells. On 32 p.n.d. the HSDH activities in the Leydig cells were observed in the control as well as in treated animals. It seems that the differentiation of peritubular Leydig cells, and thereby the steroid production, is delayed by CA, but not entirely blocked.
01 Jan 2009
TL;DR: In this paper, a pharmacophore model based on the crystal structure of the GA-derivative carbenoxolone in complex with human 11-hydroxysteroid dehydrogenase (HSD1) was used to compare the biological activity of 18-GA and its diastereomer 18GA against the two enzymes in lysates of transfected HEK-293 cells.
Abstract: Elevated cortisol concentrations have been associated with metabolic diseases such as diabetes type 2 and obesity. 11-hydroxysteroid dehydrogenase (11-HSD) type 1, catalyzing the conversion of inactive 11-ketoglucocorticoids into their active 11-hydroxy forms, plays an important role in the regulation of cortisol levels within specific tissues. The selective inhibition of 11-HSD1 is currently considered as promising therapeutic strategy for the treatment of metabolic diseases. In recent years, natural compound-derived drug design has gained considerable interest. 18-glycyrrhetinic acid (GA), a metabolite of the natural product glycyrrhizin, is not selective and inhibits both 11-HSD1 and 11-HSD2. Here, we compare the biological activity of 18-GA and its diastereomer 18-GA against the two enzymes in lysates of transfected HEK-293 cells and show that 18-GA selectively inhibits 11-HSD1 but not 11HSD2. This is in contrast to 18-GA, which preferentially inhibits 11-HSD2. Using a pharmacophore model based on the crystal structure of the GA-derivative carbenoxolone in complex with human 11HSD1, we provide an explanation for the differences in the activities of 18-GA and 18-GA. This model will be used to design novel selective derivatives of GA.
Patent•
31 Jul 2009
TL;DR: In this article, the authors used 4-phenylthiosemicarbazones of salicylic and 2-hydroxy-1-naphthoic aldehydes as inhibitors of 17β-HSD (hydroxysteroid dehydrogenase) enzyme, stimulator for prostate cancer cell multiplication.
Abstract: The invention relates to chemistry, namely to organic compounds from the class of thiosemicarbazones that can be used for treatment of prostate cancer.Summary of the invention consists in using 4-phenylthiosemicarbazones of salicylic and 2-hydroxy-1-naphthoic aldehydes as inhibitors of 17β-HSD (hydroxysteroid dehydrogenase) enzyme, stimulator for prostate cancer cell multiplication.
TL;DR: Kinetic analysis and molecular-modelling studies of 17alpha- and 17beta-hydroxysteroid substrates in the active site of AKR1C21 suggested that Gly225 and Gly226 play an important role in determining the substrate stereospecificity of the enzyme, and it is shown that these residues are critical for the binding of substrates.
Abstract: 3(17)alpha-Hydroxysteroid dehydrogenase (AKR1C21) is a unique member of the aldo-keto reductase (AKR) superfamily owing to its ability to reduce 17-ketosteroids to 17alpha-hydroxysteroids, as opposed to other members of the AKR family, which can only produce 17beta-hydroxysteroids. In this paper, the crystal structure of a double mutant (G225P/G226P) of AKR1C21 in complex with the coenzyme NADP(+) and the inhibitor hexoestrol refined at 2.1 A resolution is presented. Kinetic analysis and molecular-modelling studies of 17alpha- and 17beta-hydroxysteroid substrates in the active site of AKR1C21 suggested that Gly225 and Gly226 play an important role in determining the substrate stereospecificity of the enzyme. Additionally, the G225P/G226P mutation of the enzyme reduced the affinity (K(m)) for both 3alpha- and 17alpha-hydroxysteroid substrates by up to 160-fold, indicating that these residues are critical for the binding of substrates.