RESUMEN
Pregnane derivatives have been studied mainly for their 5α-reductase activity. However, the anti-inflammatory activities of such compounds are still poorly explored. In the search for new anti-inflammatory agents, seven new pregnane derivatives 6a-g, with cinnamic acid esters at C-3 were prepared and fully characterized. The anti-inflammatory activity of compounds was assessed in TPA induced mice ear model. From them, compound 6 b was the most active to reduce edema, with an ED50 of 0.017 mg/ear. Also, Molecular Docking and Molecular Dynamics studies were performed to identify a potential molecular target related to the inflammatory process. The in vivo results suggest that 6 b could be a potent anti-inflammatory compound, while in silico studies suggest its interaction with some critical enzymes in the inflammatory response.
Asunto(s)
Antiinflamatorios , Edema , Ratones , Animales , Simulación del Acoplamiento Molecular , Antiinflamatorios/farmacología , Edema/tratamiento farmacológico , Simulación de Dinámica Molecular , Pregnanos/uso terapéutico , Relación Estructura-ActividadRESUMEN
It is known that the growth of prostate metastatic bone tumor depends on androgens, and tumor formation can start from migratory malignant cells produced in that organ. These cells exhibit grater type 1 5α-reductase (5α-R1) activity than type 2 5α-reductase. Noteworthy, both isozymes convert testosterone (T) to the more active androgen dihydrotestosterone (DHT) in the target tissues. Thus, in order to potentially improve the prognosis of this disease, in this work, seven derivatives of 17-(1H-benzimidazol-1-yl)-16-formillandrosta-5,16-dien-3ß-yl benzoate (4a-f) and 17-(1H-benzimidazol-1-yl)-3-hydroxy-16-formylandrost-5,16-diene (4) were synthesized, characterized and identified as inhibitors of type 1 5α-reductase (5αR1). These derivatives having the advantage of improved plasma half-life. The inhibitory activity of the compounds towards 5α-R1 isoenzyme was determined by conversion of T into DHT in the presence or absence of compounds 4, 4a-f. Further, in vivo experiments were also carried out, treating gonadectomized hamsters with T and/or 4, 4a-f and evaluating their effect on the diameter of hamster flank organs and on the weight of the prostatic and seminal vesicles. Results indicated that compounds 4, 4b, 4c, served as in vitro inhibitors of the enzyme 5α-R1 and pharmacological experiments showed that 4 and derivatives 4a-f decreased the diameter of the flank glands, the weight of the prostate and seminal vesicles of treated hamsters without any appreciable toxicity during observation. Noteworthy the fact that compound 4 is the product, in all cases, of the hydrolysis of the series of esters 4a-f, thus they can serve as precursors (prodrugs) of the active form 4.
Asunto(s)
Inhibidores de 5-alfa-Reductasa/farmacología , Benzoatos/farmacología , Colestenona 5 alfa-Reductasa/metabolismo , Inhibidores de 5-alfa-Reductasa/administración & dosificación , Inhibidores de 5-alfa-Reductasa/química , Animales , Benzoatos/administración & dosificación , Benzoatos/química , Cricetinae , Relación Dosis-Respuesta a Droga , Inyecciones Subcutáneas , Microsomas Hepáticos/química , Microsomas Hepáticos/metabolismo , Conformación Molecular , Ratas , Solubilidad , Relación Estructura-ActividadRESUMEN
Pregnane derivatives are studied as agents for the treatment of different hormone-dependent diseases. The biological importance of these steroids is based on their potential use against cancer. In this study, we report the synthesis, characterization and biological activity of two pregnane derivatives with a triazole (3ß-hydroxy-21-(1H-1,2,4-triazol-1-yl)pregna-5,16-dien-20-one; T-OH) or imidazole (3ß-hydroxy-21-(1H-imidazol-1-yl)pregna-5,16-dien-20-one; I-OH) moieties at C-21. These derivatives were synthesized from 16-dehydropregnenolone acetate. The activity on cell proliferation of the compounds was measured on three human cancer cells lines: prostate cancer (PC-3), breast cancer (MCF7) and lung cancer (SK-LU-1). The cytotoxic and antiproliferative effects of T-OH and I-OH were assessed by using SBR and XTT methods, respectively. The gene expressions were evaluated by real time PCR. In addition, results were complemented by docking studies and transactivation assays using an expression vector to progesterone and androgen receptor. Results show that the two compounds inhibited the three cell lines proliferation in a dose-dependent manner. Compound I-OH downregulated the gene expression of the cyclins D1 and E1 in PC-3 and MFC7 cells; however, effect upon Ki-67, EAG1, BIM or survivin genes was not observed. Docking studies show poor interaction with the steroid receptors. Nevertheless, the transactivation assays show a weak antagonist effect of I-OH on progesterone receptor but not androgenic or antiandrogenic actions. In conclusion, the synthesized compounds inhibited cell proliferation as well as genes key to cell cycle of PC-3 and MCF7 cell lines. Therefore, these compounds could be considered a good starting point for the development of novel therapeutic alternatives to treat cancer.
Asunto(s)
Antineoplásicos/síntesis química , Imidazoles/síntesis química , Pregnadienos/síntesis química , Triazoles/síntesis química , Antineoplásicos/farmacología , Apoptosis , Proliferación Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Células HeLa , Humanos , Imidazoles/farmacología , Concentración 50 Inhibidora , Simulación del Acoplamiento Molecular , Pregnadienos/farmacología , Triazoles/farmacología , Vitamina D3 24-Hidroxilasa/metabolismoRESUMEN
Inhibitors of the enzyme 5[Formula: see text]-reductase (5aR) are promising therapeutic agents for the treatment of benign prostatic hyperplasia (BPH) and prostate cancer. The lack of structural data of the enzyme 5aR prompts the application of ligand-based approaches to systematically explore the activity landscape of 5aR inhibitors. As part of an effort to develop inhibitors of this enzyme for the treatment of BPH, herein we discuss a chemoinformatic-based analysis of the activity landscape of a novel set of 53 novel pregnane and androstene compounds. It was found that, in general, for each pair of compounds in the set, as the structure similarity of the compounds increases the corresponding potency difference decreases. These results are in agreement with an overall smooth activity landscape. However, two potent activity cliff generators were identified pointing to specific small structural changes that have a large impact on the inhibition of 5aR.
Asunto(s)
Inhibidores de 5-alfa-Reductasa/química , Androstenos/química , Pregnanos/química , Inhibidores de 5-alfa-Reductasa/farmacología , Androstenos/farmacología , Bases de Datos de Compuestos Químicos , Humanos , Masculino , Estructura Molecular , Pregnanos/farmacología , Hiperplasia Prostática/tratamiento farmacológico , Hiperplasia Prostática/enzimología , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/enzimología , Relación Estructura-ActividadRESUMEN
The enzyme type 5 17ß-hydroxysteroid dehydrogenase 5 (17ß-HSD5) catalyzes the transformation of androstenedione (4-dione) to testosterone (T) in the prostate. This metabolic pathway remains active in cancer patients receiving androgen deprivation therapy. Since physicians seek to develop advantageous and better new treatments to increase the average survival of these patients, we synthesized several different dehydroepiandrosterone derivatives. These compounds have a pyrazole or imidazole function at C-17 and an ester moiety at C-3 and were studied as inhibitors of 17ß-HSD5. The kinetic parameters of this enzyme were determined for use in inhibition assays. Their pharmacological effect was also determined on gonadectomized hamsters treated with Δ(4)-androstenedione (4-dione) or testosterone (T) and/or the novel compounds. The results indicated that the incorporation of a heterocycle at C-17 induced strong 17ß-HSD5 inhibition. These derivatives decreased flank organ diameter and prostate weight in castrated hamsters treated with T or 4-dione. Inhibition of 17ß-HSD5 by these compounds could have therapeutic potential for the treatment of prostate cancer and benign prostatic hyperplasia.
Asunto(s)
17-Hidroxiesteroide Deshidrogenasas/antagonistas & inhibidores , Deshidroepiandrosterona/farmacología , Inhibidores Enzimáticos/farmacología , Imidazoles/farmacología , Pirazoles/farmacología , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Cricetinae , Deshidroepiandrosterona/química , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Humanos , Imidazoles/química , Masculino , Persona de Mediana Edad , Estructura Molecular , Próstata/enzimología , Pirazoles/química , Relación Estructura-ActividadRESUMEN
Hyperplasia of the prostate gland and prostate cancer have been associated with high levels of serum 5α-dihydrotestosterone. This steroid is formed from testosterone by the activity of the enzyme 5α-reductase (5α-R) present in the prostate. Thus, inhibition of this enzyme could be a goal for therapies to treat these diseases. This study reports the synthesis and effects of five different 21-esters of pregnenolone derivatives as inhibitors of 5α-R types 1 and 2. The activity of these steroidal compounds was determined using in vivo and in vitro experiments. The results indicate that of the five steroids studied, the 21(p-fluoro)benzoyloxypregna-4,16-diene-3,6,20-trione derivative, whose structure has not yet been reported, has the best molecular conformation to inhibit the in vitro activity of both types of 5α-R. In addition, this steroid also displayed activity in vivo. Apparently, its pharmacological effect was increased by the presence of a keto group at C-6, because this group decreased the possibility that the steroid would be metabolized by hepatic enzymes. In addition, the double bond present at C-4 of this compound also enhanced its inhibitory activity on 5α-R, and the C-21 ester moiety increased its liphophilicity. Therefore, its solubility in the cell membrane and its pharmacological activity were both increased.
RESUMEN
It is well known that testosterone (T) under the influence of 5α-reductase enzyme is converted to dihydrotestosterone (DHT), which causes androgen-dependent diseases. The aim of this study was to synthesize new dehydroepiandrosterone derivatives (3a-e, 4a-i, 6 and 7) having potential inhibitory activity against the 5α-reductase enzyme. This paper also reports the in vivo pharmacological effect of these steroidal molecules. The results from this study showed that all compounds exhibited low inhibitory activity for 5α-reductase type 1 and 2 enzymes and they failed to bind to the androgen receptor. Furthermore, in the in vivo experiment, steroids 3b, 4f, and 4 g showed comparable antiandrogenic activity to that of finasteride; only derivatives 4d and 7 produced a considerable decrease in the weight of the prostate gland of gonadectomized hamsters treated with (T). On the other hand, compounds 4a, f and h showed 100% inhibition of the growth of prostate cancer cell line PC-3, with compound 4 g having a 98.2% antiproliferative effect at 50 µM. The overall data indicated that these steroidal molecules, having an aromatic ester moiety at C-3 (4f-h), could have anticancer properties.
Asunto(s)
Antineoplásicos/química , Antineoplásicos/farmacología , Colestenona 5 alfa-Reductasa/antagonistas & inhibidores , Deshidroepiandrosterona/análogos & derivados , Deshidroepiandrosterona/farmacología , Neoplasias de la Próstata/tratamiento farmacológico , Animales , Línea Celular Tumoral , Células Cultivadas , Colestenona 5 alfa-Reductasa/metabolismo , Cricetinae , Femenino , Humanos , Isoenzimas/antagonistas & inhibidores , Isoenzimas/metabolismo , Masculino , Ratones , Persona de Mediana Edad , Próstata/efectos de los fármacos , Próstata/metabolismo , Neoplasias de la Próstata/metabolismo , RatasRESUMEN
Abiraterone acetate is a potent inhibitor of human cytochrome P450c17 (CYP17A1, 17α-hydroxylase/17,20-lyase) and is clinically used in combination with prednisone for the treatment of castration-resistant prostate cancer. Although many studies have documented the potency of abiraterone (Abi) in a variety of in vitro and in vivo systems for several species, the exact potency of Abi for human CYP17A1 enzyme has not yet been determined, and the structural requirements for high-potency steroidal azole inhibitors are not established. We synthesized 4 Abi analogs differing in the A-B ring substitution patterns: 3α-hydroxy-Δ(4)-Abi (13), 3-keto-Δ(4)-Abi (11), 3-keto-5α-Abi (6), and 3α-hydroxy-5α-Abi (5). We measured the spectral binding constants (Ks) using purified and modified human CYP17A1 along with the determination constants (Ki) applying a native human CYP17A1 enzyme in yeast microsomes for these compounds as well as for ketoconazole. For Abi, 3-keto-Δ(4)-Abi, 3-keto-5α-Abi, and 3α-hydroxy-5α-Abi, the type 2 spectral changes gave the best fit for a quadratic equation, since in these experiments Ks values were 0.1-2.6nM, much lower than that for ketoconazole and 3α-hydroxy-Δ(4)-Abi (Ks values were 140 and 1660nM, respectively). Inhibition experiments showed mixed inhibition patterns with Ki values of 7-80nM. Abi dissociation from the CYP17A1-Abi complex was incomplete and slow; the t1/2 for dissociation was 1.8h, with 55% of complex remaining after 5h. We conclude that Abi and the 3 related steroidal azoles (3-keto-Δ(4)-Abi, 3-keto-5α-Abi, and 3α-hydroxy-5α-Abi), which also mimic natural substrates, are extraordinarily potent inhibitors of human CYP17A1, whereas the 3α-hydroxy-Δ(4)-Abi is moderately potent and comparable to ketoconazole.
Asunto(s)
Androstenoles/farmacología , Azoles/química , Inhibidores Enzimáticos/farmacología , Esteroide 17-alfa-Hidroxilasa/antagonistas & inhibidores , Esteroides/química , Androstenos , Inhibidores Enzimáticos/química , Humanos , Estructura Molecular , Esteroide 17-alfa-Hidroxilasa/metabolismo , Relación Estructura-ActividadRESUMEN
Flank organs are an androgen-dependent pilosebaceous complex present in male and female hamsters. These organs have been used for the evaluation of antiandrogenic drugs, which could be used for the treatment of androgen-dependent afflictions. This study demonstrated the role of four different steroidal carbamates 7-10 in the expression of mRNAs coding for different enzymes involved in the lipid metabolism in flank organs. To determine the biological effects of compounds 7-10 on the expression of mRNA coding for lipid enzymes, steroids 7-10, testosterone (T), progesterone (P), and/or 7-10 were applied on the flank organs. Later, the mRNA expression for the enzymes was determined by polymerase chain reaction. The binding of 8 and 9 to the progesterone receptor (PR) as well as their effects on the activity of 5α-reductase were also evaluated. Treatments with T, P, and 7-10 increased the mRNA expression for glycerol 3-phosphate acyl transferase (GPAT), ß-hydroxy-ß-methylglutaryl-CoA synthase (HMG-CoA-S), ß-hydroxy-ß-methylglutaryl-CoA reductase (HMG-CoA-R), phosphatidylinositol synthase (PI-S), and squalene-synthase (SQ-S). However, the combined treatments with P + 7-10 decreased the expression of GPAT, HMG-CoA-S, and HMG-CoA-R. Expression of mRNA for all enzymes was variable under treatment with T + 7-10. Data showed that these carbamates did not bind to the PR, but inhibited the activity of 5α-reductase. Carbamates 7-10 changed the mRNA expression model induced by T and P in flank organs.
Asunto(s)
Carbamatos/farmacología , ARN Mensajero/genética , Glándulas Sebáceas/efectos de los fármacos , Glándulas Sebáceas/enzimología , Esteroides/farmacología , 3-Oxo-5-alfa-Esteroide 4-Deshidrogenasa/metabolismo , Animales , Unión Competitiva , CDP-Diacilglicerol-Inositol 3-Fosfatidiltransferasa/genética , Carbamatos/química , Cricetinae , Farnesil Difosfato Farnesil Transferasa/genética , Femenino , Glicerol-3-Fosfato O-Aciltransferasa/genética , Humanos , Hidroximetilglutaril-CoA Reductasas/genética , Masculino , Mesocricetus , Estructura Molecular , Ovariectomía , Próstata/efectos de los fármacos , Próstata/enzimología , ARN Mensajero/metabolismo , Conejos , Receptores de Progesterona/metabolismo , Piel/efectos de los fármacos , Piel/enzimología , Esteroides/químicaRESUMEN
The aim of this study was to determine the cytotoxic effect of human cancer cells on three series of novel dehydroepiandrosterone derivatives containing triazole or pyrazole rings at C-17 and an ester moiety at C-3 of the androstane skeleton. The panel cancer cells used in this study were the following: PC-3, MCF-7 and SKLU-1. The results from this study indicated that the steroidal derivatives 4a-4e and 4f-4k showed the highest cytotoxic potency. This difference in this activity could be attributed to the ability of the triazole (three nitrogen atoms) to form stronger hydrogen bonds with the active site of the cell as compared to the pyrazole group having two nitrogen atoms. Compounds 4f-4k having an aromatic ester at C-3 showed an enhanced cytotoxic activity as compared to their aliphatic counterparts 4a-4e. Apparently the electronegative phenyl ring increased the polarity of the molecule, thus increasing the dipole-dipole association of the steroidal molecule with the reactive site of the cell.
Asunto(s)
Deshidroepiandrosterona/toxicidad , Neoplasias/tratamiento farmacológico , Antineoplásicos/síntesis química , Antineoplásicos/química , Antineoplásicos/toxicidad , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Deshidroepiandrosterona/síntesis química , Deshidroepiandrosterona/química , Humanos , Relación Estructura-ActividadRESUMEN
The aim of these studies was to synthesize twelve ester derivatives of dehydroepiandrosterone with therapeutic potential. The effect of 1-12 was demonstrated in the flank organs of gonadectomized hamsters treated with testosterone and the synthesized steroids. In vitro studies were carried out determining the IC50 values for the inhibition of the activity of 5α-reductase type 1 and 2, which are present in rat liver and human prostate respectively. The binding of 1-12 to the androgen receptors (AR) was determined using rat's prostate cytosol. Steroids 1-12 containing different substituents in the phenyl group of the ester moiety in C-3 reduced the flank organs and inhibited the activity of 5α-R type 1; however only steroids 1 and 2 inhibited 5α-R type 2. 1-12 did not bind to the AR. The modification of one atom of the substituents in the phenyl group of the ester moiety in C-3 changed their biological potency (IC50).
Asunto(s)
Inhibidores de 5-alfa-Reductasa/farmacología , Androstenos/farmacología , Colestenona 5 alfa-Reductasa/antagonistas & inhibidores , Inhibidores de 5-alfa-Reductasa/síntesis química , Inhibidores de 5-alfa-Reductasa/química , Androstenos/síntesis química , Androstenos/química , Animales , Colestenona 5 alfa-Reductasa/metabolismo , Relación Dosis-Respuesta a Droga , Humanos , Concentración de Iones de Hidrógeno , Hígado/enzimología , Masculino , Mesocricetus , Persona de Mediana Edad , Estructura Molecular , Próstata/enzimología , Ratas , Relación Estructura-ActividadRESUMEN
The effect of several new dihydroepiandrosterone ester derivatives A2-A6 was demonstrated using female cycling mice, which were synchronized for estrus with luteinizing hormone-releasing hormone (LHRH) and injected with the steroids. The binding to the progesterone receptor (PR), was obtained from the cytosol of uteri from adult estrogen-primed rabbits. A1 binds to the PR and inhibited the ovulation in cycling mice stimulated with LHRH. The activity of the endometrium and mammary glands in these mice was markedly reduced as compared to the control. A2, A4, and A5 were not active; nevertheless, A3 binds to the PR with high affinity. However, this steroid did not produce any effect as compared to that observed for the control in the endometrial and mammary glands. A6 binds to the PR with the highest affinity and induces a synergistic activity with progesterone in these tissues. Furthermore, A6 inhibited the ovulation in the same manner as A1. These results suggested that A1 and A6 are blocking the gonadotropin secretion. A1 inhibited the conversion of progesterone to 5α-progesterone. As a result of this, a blockage of the ductal and alveolar epithelial cell proliferation in the mammary and endometrial glands, which depends on 5α-progesterone, was also observed.
Asunto(s)
Androstenos/farmacología , Ciclo Estral/fisiología , Ovulación/efectos de los fármacos , Receptores de Progesterona/metabolismo , Androstenos/síntesis química , Androstenos/química , Animales , Proliferación Celular/efectos de los fármacos , Endometrio/efectos de los fármacos , Endometrio/metabolismo , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Femenino , Hormona Liberadora de Gonadotropina/administración & dosificación , Humanos , Masculino , Glándulas Mamarias Animales/efectos de los fármacos , Glándulas Mamarias Animales/metabolismo , Ratones , Persona de Mediana Edad , Progesterona/metabolismo , Alveolos Pulmonares/citología , Alveolos Pulmonares/efectos de los fármacos , Conejos , Relación Estructura-ActividadRESUMEN
BACKGROUND: The initial step of steroidal action on target cells is gene activation; therefore, the quantification of mRNA is a direct method for comparing the role of different steroids in the skin. OBJECTIVE: This study demonstrated the role of several steroids on the mRNA expression encoding for different enzymes involved in the lipid metabolism in hamsters' flank organs, which are a pilosebaceous complex. METHODS: To determine the effect of treatments with testosterone (T) progesterone (P), levonorgestrel (LNG), 17α-p-chlorobenzoyloxy-6-chloropregn-4,6-diene-3,20-dione (5) and 17α-p-chlorobenzoyloxy-4,6-pregnadiene-3,20-dione (6); T and/or LNG; T and 5 or 6; P and/or 5 or 6 on the expression of mRNA encoding for lipid enzymes, the steroids were applied to the glands; later, the mRNAs expression for the enzymes was determined by PCR. The binding of 5 and 6 to the progesterone receptor (PR) was also evaluated. RESULTS: Treatments with T, LNG, T+LNG, P, T+P, 5, T+5, T+6, P, P+5 and P+6 increased the mRNA expression for glycerol 3-phosphate acyl transferase (GPAT), ß-hydroxy-ß-methylglutaryl-CoA synthase (HMG-CoA-S), ß-hydroxy-ß-methylglutaryl-CoA reductase (HMG-CoA-R), phosphatidylinositol synthase as compared to the controls. However, squalene synthase was increased with all treatments except with T+5 and 6; 6 did not significantly increase the expression for GPAT or HMG-CoA-S, however it increased the concentration of HMG-CoA-R enzyme. 5 and 6 bind to the PR, thus indicating that the effect of these steroids on the mRNA expression could be the result of their binding. CONCLUSION: The lipid metabolism is regulated by several steroids thought different mechanism of action, in flank organs.
Asunto(s)
Glándulas Sebáceas/efectos de los fármacos , Glándulas Sebáceas/metabolismo , Esteroides/farmacología , Animales , Secuencia de Bases , CDP-Diacilglicerol-Inositol 3-Fosfatidiltransferasa/genética , Cricetinae , Cartilla de ADN/genética , Farnesil Difosfato Farnesil Transferasa/genética , Femenino , Expresión Génica/efectos de los fármacos , Glicerol-3-Fosfato O-Aciltransferasa/genética , Hidroximetilglutaril-CoA Reductasas/genética , Levonorgestrel/farmacología , Metabolismo de los Lípidos/efectos de los fármacos , Metabolismo de los Lípidos/genética , Mesocricetus , Progesterona/farmacología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Progesterona/metabolismo , Piel/efectos de los fármacos , Piel/metabolismo , Esteroides/química , Esteroides/metabolismo , Testosterona/farmacologíaRESUMEN
The aim of this study was to synthesize different ester derivatives of dehydroepiandrosterone with therapeutic potential as antiandrogens. The biological effect of these steroids was demonstrated in in vivo as well as in vitro experiments. In the in vivo experiments, we measured the activity of seven steroids on the weight of the prostate and seminal vesicles of gonadectomized hamsters treated with testosterone. For the in vitro studies, we determined the IC(50) values by measuring the concentration of the steroidal derivatives that inhibits 50% of the activity of 5α-reductase present in human prostate and also its binding capacity to the androgen receptors (AR) obtained from rat's prostate cytosol. The results from these experiments indicated that compounds 7 5α,6ß-dibromo-3ß-propanoyloxyandrostan-17-one, 8 5α,6ß-dibromo-3ß-butanoyloxyandrostan-17-one and 9 5α,6ß-dibromo-3ß-(3'-oxapentanoyloxy)-androstan-17-one, significantly decreased the weight of the prostate and seminal vesicles as compared to testosterone treated animals; this reduction of the weight of these glands was comparable to that produced by Finasteride 11. On the other hand, compounds 4 3ß-acetoxyandrost-5-en-17-one, 5 3ß-hexanoyloxyandrost-5-en-17-one 6 3ß-(3'-oxapentanoyloxy)-androst-5-en-17-one, 7 and 12 dehydroepiandrosterone, (commercially available) inhibited the enzyme 5α-reductase. Compounds 4, 5, 6, 8 and 9 (IC(50) values of 5.2±1.2, 0.049±0.002, 6.4±1.1, 0.10±0.045, and 6.8±0.9 nM, respectively) exhibited the highest inhibitory activity. However, none of these compounds binds to the AR.
Asunto(s)
Inhibidores de 5-alfa-Reductasa/síntesis química , Antagonistas de Andrógenos/síntesis química , Colestenona 5 alfa-Reductasa/metabolismo , Deshidroepiandrosterona/análogos & derivados , Animales , Cricetinae , Humanos , Masculino , Mesocricetus , RatasRESUMEN
This study reports the synthesis of several new steroidal lactones: 5α,6ß-dibromo-17a-oxa-D-homoandrostane-3ß-yl-3'-oxapentanoate (11), 5α,6ß-dibromo-17a-oxa-D-homoandrostane-3ß-yl-propanoate (12), 5α,6ß-dibromo-17a-oxa-D-homoandrostane-3ß-yl-butanoate (13), 5α,6ß-dibromo-17a-oxa-D-homoandrostane-3ß-yl-pentanoate (14), 5α,6ß-dibromo-17a-oxa-D-homoandrostane-3ß-yl-hexanoate (15), 17a-oxa-D-homoandrost-5-en-17-one-3ß-yl-3'-oxapentanoate (16), 17a-oxa-D-homoandrost-5-en-17-one-3ß-yl-propanoate (17), 17a-oxa-D-homoandrost-5-en-17-one-3ß-yl-butanoate (18), 17a-oxa-D-homoandrost-5-en-17-one-3ß-yl-pentanoate (19) and 17a-oxa-D-homoandrost-5-en-17-one-3ß-yl-hexanoate (20) with a therapeutic potential as antiandrogens. The biological effect of these steroids was demonstrated in in vivo as well as in vitro experiments. In the in vivo experiments, we measured the activity of ten new steroidal derivatives on the weight of the prostate and seminal vesicle glands of gonadectomized hamsters treated with testosterone. For the in vitro studies, we determined the IC(50) values by measuring the concentration of the steroidal derivatives that inhibits 50% of the activity of the 5α-reductase enzyme present in human prostate and also its binding capacity to the androgen receptors (AR) obtained from rat's prostate cytosol. The results from these experiments indicated that compounds 11-20, significantly decreased the weight of the prostate and seminal vesicles as compared to testosterone treated animals; this reduction of the weight of these glands was comparable to that produced by Finasteride. On the other hand, compounds 11-20 inhibited the enzyme 5α-reductase, with compounds 14-19 (IC(50) values of 4.2 ± 0.95, 0.025 ± 0.003, 1.2 ± 0.45, 1.2 ± 0.1, 0.028 ± 0.003, and 0.069 ± 0.005 nM, respectively) showing the highest inhibitory activity. The results from the in vitro experiments indicated that only 15-17 bind to the AR.
Asunto(s)
Inhibidores de 5-alfa-Reductasa/farmacología , Lactonas/farmacología , Receptores Androgénicos/efectos de los fármacos , Esteroides/farmacología , Concentración 50 Inhibidora , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Espectrofotometría InfrarrojaRESUMEN
The aim of this study was to determine the capacity of some progesterone derivatives, to inhibit the conversion of labeled androstenedione ([(3)H] 4-dione) to [(3)H]dihydrotestosterone ([(3)H]DHT) in prostate nuclear membrane fractions, where the 5α-reductase activity is present. The enzyme 5α-reductase catalyzes the 5α-reduction of 4-dione whereas the 17ß-hydroxysteroid dehydrogenase catalyzes the transformation of 4-dione to testosterone or 5α-dione to dihydrotestosterone (DHT). Moreover, we also investigated the role of unlabeled 5α-dione in these pathways. In order to determine the inhibitory effect of different concentrations of the progesterone derivatives in the conversion of [(3)H] 4-dione to [(3)H]DHT, homogenates of human prostate were incubated with [(3)H] 4-dione, NADPH and increasing concentrations of non-labeled 5α-dione. The incubating mixture was extracted and purified using thin layer chromatography. The fraction of the chromatogram corresponding to the standard of DHT was separated and the radioactivity determined. The results showed that the presence of [(3)H] 4-dione plus unlabelled 5α-dione produced similar levels of DHT as compared to [(3)H] 4-dione. On the other hand, the results indicated that 17α-hydroxypregn-4-ene-3,20-dione 5 and 4-bromo-17α-hydroxypregn-4-ene-3,20-dione 7b, were the most potent steroids to inhibit the conversion of [(3)H] 4-dione to [(3)H]DHT, showing IC(50) values of 2 and 1.6 nM, respectively.
Asunto(s)
Inhibidores de 5-alfa-Reductasa/química , Inhibidores de 5-alfa-Reductasa/farmacología , Androstenodiona/química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Adulto , Androstenodiona/antagonistas & inhibidores , Androstenodiona/metabolismo , Activación Enzimática/efectos de los fármacos , Humanos , Concentración 50 Inhibidora , Masculino , Estructura Molecular , Membrana Nuclear/enzimología , Próstata/metabolismoRESUMEN
The aim of this study was to ascertain the inhibitory effect of several progesterone derivatives for 5 alpha-reductase types 1 and 2 isozymes and to determine the binding to the androgen receptor. The 3,20-dioxopregna-4-ene-17 alpha-yl acetate 4 containing an acetoxy group in C-17 and steroid 17 alpha-hydroxypregn-4-ene-3,20-dione 5 having a hydroxyl group in the same position inhibited both isozymes. On the other hand, 17 alpha-hydroxy-4,5-epoxypregnan-3,20-dione 6 with an epoxy function at C-4, inhibited only the type 1 enzyme. Steroid 4-chloro-17 alpha-hydroxypregn-4-ene-3,20-dione 7a and 4-bromo-17 alpha-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 5 alpha-reductase. These results indicate that an increase in the electronegativity of ring A produces a major inhibitory activity for 5 alpha-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-17 alpha yl-4-ethylbenzoate 8a and 3,20-dioxo-4-bromopregn-4-ene-17 alpha yl-4-ethylbenzoate 8b were obtained; these steroids inhibited only the 5 alpha-reductase type 2 enzyme. Finasteride and steroids 4, 5, 7b, 8a showed a comparable in vivo pharmacological activity, however the IC(50) 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.
Asunto(s)
3-Oxo-5-alfa-Esteroide 4-Deshidrogenasa/metabolismo , Progesterona/análogos & derivados , Progesterona/metabolismo , Receptores Androgénicos/metabolismo , Inhibidores de 5-alfa-Reductasa , Animales , Unión Competitiva , Cricetinae , Inhibidores Enzimáticos/farmacología , Finasterida/metabolismo , Finasterida/farmacología , Humanos , Isoenzimas/antagonistas & inhibidores , Isoenzimas/metabolismo , Cinética , Masculino , Mesocricetus , Persona de Mediana Edad , Estructura Molecular , Tamaño de los Órganos/efectos de los fármacos , Progesterona/farmacología , Próstata/efectos de los fármacos , Próstata/enzimología , Próstata/crecimiento & desarrollo , Unión Proteica , Vesículas Seminales/efectos de los fármacos , Vesículas Seminales/crecimiento & desarrolloRESUMEN
The aim of this study was to synthesize three different D-homoandrostadiene derivatives (2-4) and study their biological activity. We carried out in vivo and in vitro experiments using female cycling mice, which were synchronized for estrus with luteinizing hormone-releasing hormone (LHRH) and injected with the steroidal compounds. It was also determined the binding of these compounds to the progesterone receptors (PR). Since these steroids have a new D-homoandrostandienone skeleton in their molecular structure, it was of interest also to study their binding to the androgen receptors (AR). After LHRH treatment, the mice of the control group showed the presence of 14+/-4 corpus lutea in the ovary whereas the animals treated with steroids 2-4, with RBAs of 100%, exhibited 11+/-7, 12+/-2, and 10+/-4 respectively. As a result of this study, it is evident that these steroids did not inhibit the ovulation in these animals. The uterus of the control group, showed the typical progestational activity with an enlarged endometrial thickness with a secretory activity. However, the endometrium of the mice treated with steroids 2-4 did not show an enlargement of the endometrium and no secretory activity could be detected. This fact indicates that compounds 2-4 had antagonistic activity in this tissue. The overall data show that steroids 2-4 are antagonists of the PR. However, they do not bind to the AR. These results also demonstrate that 2-4 have an antiprogestational activity in vivo, but do not decrease the number of corpus lutea in the ovary of mice treated with LHRH.
Asunto(s)
Androstadienos/química , Androstadienos/farmacología , Receptores de Progesterona/antagonistas & inhibidores , Antagonistas de Receptores Androgénicos , Androstadienos/metabolismo , Androstenodiona/análogos & derivados , Androstenodiona/química , Androstenodiona/farmacología , Animales , Unión Competitiva , Cuerpo Lúteo/efectos de los fármacos , Cuerpo Lúteo/metabolismo , Endometrio/efectos de los fármacos , Endometrio/metabolismo , Femenino , Hormona Liberadora de Gonadotropina/farmacología , Homoesteroides/química , Homoesteroides/farmacología , Antagonistas de Hormonas/química , Antagonistas de Hormonas/metabolismo , Antagonistas de Hormonas/farmacología , Masculino , Ratones , Mifepristona/química , Mifepristona/metabolismo , Mifepristona/farmacología , Estructura Molecular , Ovario/efectos de los fármacos , Ovario/metabolismo , Fenilacetatos/química , Progesterona/química , Progesterona/metabolismo , Progesterona/farmacología , Conejos , Ratas , Receptores Androgénicos/metabolismo , Receptores de Progesterona/metabolismo , Relación Estructura-Actividad , Útero/efectos de los fármacos , Útero/metabolismoRESUMEN
The present study is addressed to ascertain the inhibitory effect of several progesterone derivatives having a chlorine substituent at C-6 (12a-12d), 15 with a bromine substituent at C-6 and 14a-14d, without any halogen atom at C-6 all having an ester side chain at C-17 (benzoate ester bearing a Cl, F and a Br atom at C-4 position of the phenyl ring) on the 5alpha-reductase enzyme activity present in human prostate. In addition, it was also of interest to investigate the pharmacological effect on hamster flank organs diameter size. In order to study the structure-activity relationships of steroids 12a-12d, 14a-14d and 15 we determined the concentration of these steroids that inhibited 50% of the activity of human prostate 5alpha-reductase enzyme (IC(50)), as well as the in vivo effect of these compounds in the hamster flank organs diameter size. We also ascertained, the capacity of these steroids to bind to the androgen receptors present in the rat prostate cytosol using labeled mibolerone (MIB) for monitoring the binding to the androgen receptor. The results from this study indicated that compounds 12a-12d (having a chlorine substituent at C-6), 14a-14d (lacking a halogen atom at C-6), 13 and 15 (having a bromine atom at C-6) showed an increased antiandrogenic effect (lower value for the diameter of the flank organs) as compared to the flank organs from testosterone-treated hamsters. On the other hand, the series of compounds containing a chlorine substituent at C-6 compounds (12a-12d) showed a higher antiandrogenic activity as compared to the compounds lacking a halogen atom at C-6 (14a, 14b and 14d). Although compounds 13 and 15 decreased the flank organs diameter size, however, this increase was not statistically significant as compared to that of the commercially available product finasteride. The steroidal derivatives 13, 14a-14d (lacking the chlorine substituent at C-6) and 15 (having a bromine atom at C-6) exhibited a higher 5alpha-reductase inhibitory activity (lower IC(50) values) as compared to the series of compounds 12a-12d having the halogen substituent at C-6. Finasteride reduced the diameter size of the flank organs. The effect of this steroid and compounds 12a-12d, 13, 14a-14d and 15 on hamster flank organs can be explained by the fact that these steroids did not bind to the androgens receptor, which indicates that its mechanism of action is an inhibiting for the 5alpha-reductase activity. This enzyme is present in the hamster flank organs and was inhibited by the novel steroids in the human prostate homogenates.
Asunto(s)
Colestenona 5 alfa-Reductasa/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Pregnanos/química , Pregnanos/farmacología , Anciano , Animales , Unión Competitiva , Colestenona 5 alfa-Reductasa/metabolismo , Cricetinae , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/metabolismo , Humanos , Concentración 50 Inhibidora , Masculino , Tamaño de los Órganos/efectos de los fármacos , Pregnanos/síntesis química , Pregnanos/metabolismo , Próstata/efectos de los fármacos , Próstata/enzimología , Ratas , Relación Estructura-ActividadRESUMEN
The aim of this study was to determine the biological activity of 4 steroidal derivatives (9a, 9b and 10a, 10b) prepared from the commercially available 17alpha acetoxyprogesterone, where 9a, 9b, have the Delta(4)-3-oxo structure and 10a and 10b an epoxy group at C-4 and C-5. These steroids were tested as inhibitors of 5alpha-reductase enzyme, which is present in androgen-dependent tissues and converts testosterone to its more active reduced metabolite dihydrotestosterone. The pharmacological effect of these steroids was demonstrated by the significant decrease of the weight of the prostate gland of gonadectomized hamsters treated with testosterone plus finasteride or with steroids 10a and 10b. For the studies in vitro the IC(50) values were determined by measuring the steroid concentration that inhibits 50% of the activity of-5alpha-reductase. In this study we also determined the capacity of these steroids to bind to the androgen receptor present in the rat prostate cytosol. The results from this work indicated that compounds 9a, 9b, 10a, and 10b inhibited the 5alpha reductase activity with IC(50) values of 360, 370, 13 and 4.9 nM respectively. However these steroids did not bind to the androgen receptors since none competed with labeled mibolerone. Steroid 10b, an epoxy steroidal derivative containing bromine atom in the ester moiety, was the most active inhibitor of 5alpha-reductase enzyme, present in human prostate homogenates with an IC(50) value of 4.9 nM and also showed in vivo pharmacological activity since it decreased the weight of the prostate from hamsters treated with testosterone in a similar way as finasteride.