RESUMEN
A novel synthetic route of producing ursodeoxycholic acid (UDCA) was developed through multiple reactions from plant-source dehydroepiandrosterone (DHEA), with a Mistunobu reaction and regioselective allyl oxidationat as the key steps. The reaction conditions of the key allyl oxidation reaction were also investigated and optimized, including solvent, oxidant and reaction temperature. In this novel route for the preparation of UDCA, most of the reaction steps have high conversions and overall yield up to 35% for 8 steps. Since all starting materials are cost-effective, commercially available and effectively avoided the risk of animal derived raw materials, this promising synthetic route offers economical and efficient strategies for potential production of UDCA.
Asunto(s)
Deshidroepiandrosterona/química , Oxidantes/química , Ácido Ursodesoxicólico/síntesis química , Oxidación-Reducción , SolventesRESUMEN
We engineered the cytochrome P450 monooxygenase CYP107D1 (OleP) from Streptomyces antibioticus for the stereo- and regioselective 7ß-hydroxylation of lithocholic acid (LCA) to yield ursodeoxycholic acid (UDCA). OleP was previously shown to hydroxylate testosterone at the 7ß-position but LCA is exclusively hydroxylated at the 6ß-position, forming murideoxycholic acid (MDCA). Structural and 3DM analysis, and molecular docking were used to identify amino acid residues F84, S240, and V291 as specificity-determining residues. Alanine scanning identified S240A as a UDCA-producing variant. A synthetic "small but smart" library based on these positions was screened using a colorimetric assay for UDCA. We identified a nearly perfectly regio- and stereoselective triple mutant (F84Q/S240A/V291G) that produces 10-fold higher levels of UDCA than the S240A variant. This biocatalyst opens up new possibilities for the environmentally friendly synthesis of UDCA from the biological waste product LCA.
Asunto(s)
Proteínas Bacterianas/metabolismo , Sistema Enzimático del Citocromo P-450/metabolismo , Ácido Ursodesoxicólico/metabolismo , Proteínas Bacterianas/genética , Sitios de Unión , Dominio Catalítico , Sistema Enzimático del Citocromo P-450/genética , Ácido Desoxicólico/química , Ácido Desoxicólico/metabolismo , Hidroxilación , Ácido Litocólico/química , Ácido Litocólico/metabolismo , Simulación del Acoplamiento Molecular , Mutagénesis Sitio-Dirigida , Estereoisomerismo , Streptomyces/enzimología , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/químicaRESUMEN
A novel synthetic route of producing ursodeoxycholic acid (UDCA) was developed through multiple reactions from cheap and commercially available bisnoralcohol (BA). The key reaction conditions, including solvents, bases and reaction temperatures of the route were investigated and optimized. In the straightforward route for preparation of UDCA, most of the reaction steps have high conversions with average yields of 91%, and overall yield up to 59% (6 steps) from the plant-source BA. Especially in the last step of reduction and hydrolysis, there are five functional groups converted with calcd 97% per conversion in one-pot reaction. This promising route offers economical and efficient strategies for potential large-scale production of UDCA.
Asunto(s)
Plantas/química , Ácido Ursodesoxicólico/síntesis química , 20-alfa-Dihidroprogesterona , Conformación Molecular , Estereoisomerismo , Ácido Ursodesoxicólico/químicaRESUMEN
A novel synthetic route of producing ursodeoxycholic acid (UDCA) and obeticholic acid (OCA) was developed through multiple reactions from cheap and readily-available cholic acid. The reaction conditions of the key elimination reaction of mesylate ester group were also investigated and optimized, including solvent, base and reaction temperature. In the straightforward synthetic route for preparation of UDCA and OCA, most of the reaction steps have high conversions with average yields of 94% and 92%, and overall yield up to 65% (7 steps) and 36% (11 steps) from cholic acid, respectively. This promising route offers economical and efficient strategies for potential large-scale production of UDCA and OCA.
Asunto(s)
Ácido Quenodesoxicólico/análogos & derivados , Ácido Cólico/química , Ácido Ursodesoxicólico/síntesis química , Técnicas de Química Sintética , Ácido Quenodesoxicólico/síntesis química , Ácido Quenodesoxicólico/química , Análisis Costo-Beneficio , Ácido Ursodesoxicólico/químicaRESUMEN
Ursodeoxycholic acid (UDCA) is considered the first-choice therapy for cholestatic disorders. To enhance solubility and exploit specific transporters in liver, we synthesized a new galactosyl pro-drug of UDCA (UDCAgal). Ethinylestradiol (EE)-induced cholestasis was used to study and compare the effects of UDCAgal with UDCA on bile flow, hepatic canalicular efflux transporter expression, and inflammation. UDCAgal resulted quite stable both at pH 7.4 and 1.2 and regenerated the parent drug after incubation in human plasma. Its solubility, higher than UDCA, was pH- and temperature-independent. UDCAgal displayed a higher cell permeation compared to UDCA in liver HepG2 cells. Moreover, in cholestatic rats, UDCAgal showed a higher potency compared to UDCA in reducing serum biomarkers (AST, ALT, and ALP) and cytokines (TNF-α and IL-1ß). The higher effect of UDCAgal on the increase in bile salt export pump and multidrug resistance-associated protein 2 transcription indicated an improved spillover of bile acids from the liver. UDCAgal showed a reduction in CCL2, as well as TNF-α, IL-1ß, and cyclooxygeanse-2 mRNAs, indicating a reduction in hepatic neutrophil accumulation and inflammation. Moreover, UDCAgal, similarly to UDCA, heightens bile flow and modulates biliary acids secretion. These results indicate that UDCAgal has a potential in the treatment of cholestatic disease.
Asunto(s)
Colestasis/tratamiento farmacológico , Estrógenos/toxicidad , Ácido Ursodesoxicólico/química , Ácido Ursodesoxicólico/uso terapéutico , Animales , Colestasis/metabolismo , Ciclooxigenasa 2/sangre , Etinilestradiol/toxicidad , Células Hep G2 , Humanos , Interleucina-1beta/sangre , Masculino , Proteína 2 Asociada a Resistencia a Múltiples Medicamentos , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/sangre , Profármacos/síntesis química , Profármacos/química , Profármacos/uso terapéutico , Ratas , Ratas Wistar , Solubilidad , Factor de Necrosis Tumoral alfa/sangre , Ácido Ursodesoxicólico/síntesis químicaRESUMEN
BACKGROUND: Bile salts are steroidal biosurfactants. Micellar systems of bile salts are not only important for solubilization of cholesterol, but they also interact with certain drugs thus changing their bioavailability. METHODS: The number-average aggregation numbers (n¯) are determined using the Moroi-Matsuoka-Sugioka thermodynamic method. Critical micellar concentrations were determined by spectrofluorometric method using pyren and by surface tension measurements. RESULTS: Micelles of ethylidene derivatives possess the following values for n¯: 7-Eth-D (n¯=11 (50 mM)-n¯=14.8 (100 mM)); 12-Ox-7-Eth-L (n¯≈8.8, without concentration dependence) and 7,12-diOx-3-Eth-Ch (n¯≈2.9, without concentration dependence). In the planes n¯-ln k and ln CMC-ln k derivative 7-Eth-D is outlier in respect to hydrophobic linear congeneric groups. CONCLUSION: Gibbs energy of formation for 7-Eth-D anion micelles in addition to the Gibbs energy of hydrophobic interactions consists excess Gibbs energy (GE) from hydrogen bond formation between building blocks of micelles. Gibbs energy of formation for 7,12-diOx-3-Eth-Ch and 12-Ox-7-Eth-L anion micelle is determined by the Gibbs energy of hydrophobic interactions. Relative increase in hydrophobicity and aggregation number for ethylidene derivatives is larger when ethylidene group is introduced from the C7 lateral side of steroidal skeleton then it is when ethylidene group is on C3 carbon. GENERAL SIGNIFICANCE: Position of outlier towards hydrophobic congeneric groups from n¯-ln k and ln CMC-ln k planes indicates the existence of excess Gibbs energy (GE) which is not of hydrophobic nature (formation of hydrogen bonds). For the bile salt micelles to have GE (formation of secondary micelles) it is necessary that steroidal skeleton possesses C3-α-(e)-OH and C12-α-(a)-OH groups.
Asunto(s)
Ácidos y Sales Biliares/química , Colesterol/química , Micelas , Esteroides/química , Algoritmos , Ácidos y Sales Biliares/síntesis química , Ácido Desoxicólico/síntesis química , Ácido Desoxicólico/química , Enlace de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Modelos Químicos , Estructura Molecular , Solubilidad , Espectrometría de Fluorescencia , Esteroides/síntesis química , Tensión Superficial , Termodinámica , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/químicaRESUMEN
Ursodeoxycholic acid (UDCA) is a bile acid with demonstrated anti-apoptotic activity in both in vitro and in vivo models. However, its utility is hampered by limited aqueous solubility. As such, water-soluble prodrugs of UDCA could have an advantage over the parent bile acid in indications where intravenous administration might be preferable, such as decreasing damage from stroke or acute kidney injury. Five phosphate prodrugs were synthesized, including one incorporating a novel phosphoryloxymethyl carboxylate (POMC) moiety. These prodrugs were highly water-soluble, but showed significant differences in chemical stability, with oxymethylphosphate prodrugs being the most unstable. In a series of NMR experiments, the POMC prodrug was bioactivated to UDCA by alkaline phosphatase (AP) faster than a prodrug containing a phosphate directly attached to the alcohol at the 3-position of UDCA. Both of these prodrugs showed significant anti-apoptotic activity in a series of in vitro assays, although the POMC prodrug required the addition of AP for activity, while the other compound was active without exogenous AP.
Asunto(s)
Apoptosis/efectos de los fármacos , Profármacos/farmacología , Ácido Ursodesoxicólico/farmacología , Agua/química , Animales , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Hepatocitos/efectos de los fármacos , Masculino , Conformación Molecular , Profármacos/síntesis química , Profármacos/química , Ratas , Solubilidad , Relación Estructura-Actividad , Factor de Crecimiento Transformador beta1/antagonistas & inhibidores , Factor de Crecimiento Transformador beta1/metabolismo , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/químicaRESUMEN
A series of N-sulfonyl-3,7-dioxo-5ß-cholan-24-amides, ursodeoxycholic acid derivatives, have been designed and synthesized in nine steps starting from ursodeoxycholic acid. The in vitro antitumor activity of the target compounds has been evaluated against HCT-116, MCF-7, K562, and SGC-7901 cell lines. The pharmacological results showed that most of the prepared compounds display excellent selective cytotoxicity toward HCT-116, MCF-7, and K562 cell lines. Particularly, compounds 10c, 10f and 10g show high inhibitory activity on these human cancer cell lines (IC50: 2.39-9.34 µM). Conversely, all compounds are generally inactive against SGC-7901, with only 10b having IC50 below 50 µM.
Asunto(s)
Antineoplásicos/síntesis química , Sulfonamidas/síntesis química , Ácido Ursodesoxicólico/análogos & derivados , Ácido Ursodesoxicólico/síntesis química , Antineoplásicos/farmacología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Concentración 50 Inhibidora , Oxidación-Reducción , Relación Estructura-Actividad , Sulfonamidas/farmacología , Ácido Ursodesoxicólico/farmacologíaRESUMEN
Deoxycholic acid (DCA), a secondary bile acid (BA), and ursodeoxycholic acid (UDCA), a tertiary BA, cause opposing effects in vivo and in cell suspensions. Fluorescent analogues of DCA and UDCA could help investigate important questions about their cellular interactions and distribution. We have prepared a set of isomeric 3α- and 3ß-amino analogues of UDCA and DCA and derivatised these with the discrete fluorophore, 4-nitrobenzo-2-oxa-1,3-diazol (NBD), forming the corresponding four fluorescent adducts. These absorb in the range 465-470 nm and fluoresce at approx. 535 nm. In order to determine the ability of the new fluorescent bile acids to mimic the parents, their uptake was studied using monolayers of Caco-2 cells, which are known to express multiple proteins of the organic anion-transporting peptide (OATP) subfamily of transporters. Cellular uptake was monitored over time at 4 and 37°C to distinguish between passive and active transport. All four BA analogues were taken up but in a strikingly stereo- and structure-specific manner, suggesting highly discriminatory interactions with transporter protein(s). The α-analogues of DCA and to a lesser extent UDCA were actively transported, whereas the ß-analogues were not. The active transport process was saturable, with Michaelis-Menten constants for 3α-NBD DCA (5) being K(m)=42.27±12.98 µM and V(max)=2.8 ± 0.4 nmol/(mg protein*min) and for 3α-NBD UDCA (3) K(m)=28.20 ± 7.45 µM and V(max)=1.8 ± 0.2 nmol/(mg protein*min). These fluorescent bile acids are promising agents for investigating questions of bile acid biology and for detection of bile acids and related organic anion transport processes.
Asunto(s)
Ácido Desoxicólico/análogos & derivados , Colorantes Fluorescentes/química , Ácido Ursodesoxicólico/análogos & derivados , Transporte Biológico , Células CACO-2 , Ácido Desoxicólico/síntesis química , Ácido Desoxicólico/química , Ácido Desoxicólico/farmacocinética , Ácido Desoxicólico/farmacología , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/farmacocinética , Humanos , Estereoisomerismo , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/química , Ácido Ursodesoxicólico/farmacocinéticaRESUMEN
Rational exploration directed by DFT (density functional theory) based atomic Fukui indices, lead to development of regioselective oxidation of cholic acid and its 7ß epimer by o-iodoxybenzoic acid. In case of cholic acid only, 7α-hydroxyl underwent oxidation, where as in its 7ß epimer the selectivity was towards 12α-hydroxy group. Since these oxidations are the key steps in synthesis of ursodeoxycholic acid starting from cholic acid these findings may be useful in devising a protection free synthetic route.
Asunto(s)
Ácido Cólico/química , Yodobencenos/química , Ácido Ursodesoxicólico/química , Espectroscopía de Resonancia Magnética , Estructura Molecular , Oxidación-Reducción , Ácido Ursodesoxicólico/síntesis químicaRESUMEN
Ursodeoxycholic acid (UDCA) is used for the treatment of hepatic inflammatory diseases. Recent studies have shown that UDCA's biological effects are partly glucocorticoid receptor (GR) mediated. UDCA derivatives were synthesized and screened for ability to induce GR translocation in a high content analysis assay using the esophageal cancer SKGT-4 cell line. UDCA derivatives induced GR translocation in a time dependent manner with equal efficacy to that of dexamethasone (Dex) and with greatly increased potency relative to UDCA. The cyclopropylamide 1a suppressed TNF-α induced NF-κB activity and it induced GRE transactivation. 1a was unable to displace Dex from the GR ligand binding domain (LBD) in a competition experiment but was capable of coactivator recruitment in a time-resolved fluorescence energy transfer assay (TR-FRET). This represents a novel mechanism of action for a GR modulator. These derivatives could result in a new class of GR modulators.
Asunto(s)
Amidas/síntesis química , Receptores de Glucocorticoides/metabolismo , Ácido Ursodesoxicólico/análogos & derivados , Ácido Ursodesoxicólico/síntesis química , Amidas/farmacología , Sitios de Unión , Unión Competitiva , Línea Celular Tumoral , Dexametasona/metabolismo , Dexametasona/farmacología , Neoplasias Esofágicas , Transferencia Resonante de Energía de Fluorescencia , Glucocorticoides/metabolismo , Glucocorticoides/farmacología , Células HEK293 , Humanos , Ligandos , Modelos Moleculares , FN-kappa B/metabolismo , Transporte de Proteínas , Ensayo de Unión Radioligante , Receptores de Glucocorticoides/agonistas , Receptores de Glucocorticoides/genética , Elementos de Respuesta , Relación Estructura-Actividad , Activación Transcripcional , Factor de Necrosis Tumoral alfa/farmacología , Ácido Ursodesoxicólico/farmacologíaRESUMEN
23-Alkyl-substituted and 6,23-alkyl-disubstituted derivatives of chenodeoxycholic acid are identified as potent and selective agonists of TGR5, a G-protein coupled receptor for bile acids (BAs). In particular, we show that methylation at the C-23(S) position of natural BAs confers a marked selectivity for TGR5 over FXR, while the 6alpha-alkyl substitution increases the potency at both receptors. The present results allow for the first time a pharmacological differentiation of genomic versus nongenomic effects mediated by BA derivatives.
Asunto(s)
Ácidos y Sales Biliares/síntesis química , Receptores Acoplados a Proteínas G/agonistas , Ácido Ursodesoxicólico/análogos & derivados , Animales , Ácidos y Sales Biliares/química , Ácidos y Sales Biliares/farmacología , Células CHO , Cricetinae , Cricetulus , Cristalografía por Rayos X , Proteínas de Unión al ADN/agonistas , Humanos , Ligandos , Metilación , Modelos Moleculares , Estructura Molecular , Receptores Citoplasmáticos y Nucleares/agonistas , Estereoisomerismo , Relación Estructura-Actividad , Factores de Transcripción/agonistas , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/química , Ácido Ursodesoxicólico/farmacologíaAsunto(s)
Sistemas de Liberación de Medicamentos , Donantes de Óxido Nítrico , Ácido Oleanólico , Animales , Apoptosis/efectos de los fármacos , Compuestos Azo/síntesis química , Compuestos Azo/farmacología , Línea Celular Tumoral , Humanos , Hígado/metabolismo , Neoplasias Hepáticas/patología , Nitratos/síntesis química , Nitratos/farmacología , Donantes de Óxido Nítrico/síntesis química , Donantes de Óxido Nítrico/farmacología , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/síntesis química , Ácido Oleanólico/farmacología , Piperazinas/síntesis química , Piperazinas/farmacología , Ácido Ursodesoxicólico/análogos & derivados , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/farmacologíaRESUMEN
The microbial 7alpha-OH epimerisation of cholic, chenodeoxycholic, and 12-ketochenodeoxycholic acids (7alpha-OH bile acids) with Xanthomonas maltophilia CBS 827.97 to corresponding 7beta-OH derivatives with scarcity of oxygen is described. With normal pressure of oxygen the 7-OH oxidation products are obtained. No biotransformations are achieved in anaerobic conditions. The microbial 7alpha-OH epimerisation is achieved by oxidation of 7-OH function and subsequent reduction. Partial purification, in fact, of the enzymatic fraction revealed the presence of two hydroxysteroid dehydrogenases (HSDH) alpha- and beta-stereospecific together with a glycocholate hydrolase. On the basis of these results a further application is the microbial reduction of 6alpha-fluoro and 6beta-fluoro-3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid methyl esters to the corresponding 7alpha-OH and 7beta-OH derivatives.
Asunto(s)
Ácidos y Sales Biliares/química , Ácido Desoxicólico/análogos & derivados , Ácido Desoxicólico/química , Stenotrophomonas maltophilia/metabolismo , Ácido Ursodesoxicólico/análogos & derivados , Ácidos y Sales Biliares/metabolismo , Ácido Desoxicólico/síntesis química , Hidroxiesteroide Deshidrogenasas/metabolismo , Modelos Químicos , Oxígeno/metabolismo , Espectrofotometría , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/químicaRESUMEN
5-Aminosalicylic acid conjugate of ursodeoxycholic acid was synthesized in above 90% yield by adding a basic solution of 5-aminosalicylic acid into the mixed anhydride formed with ursodeoxycholic acid and ethyl chloroformate. The 5-aminosalicylic acid conjugate of ursodeoxycholic acid was poorly secreted into the bile and was deconjugated with cholylglycine hydrolase and Clostridium perfringens, that deconjugate naturally occurring glycine and taurine conjugates of bile acids. However, ursodeoxycholic acid 5-aminosalicylic acid conjugate was not absorbed from the duodenum but was concentrated in the colon where it was partially hydrolyzed by the intestinal bacteria to ursodeoxycholic acid and 5-aminosalicylic acid. We believe that this unique conjugation of ursodeoxycholic acid with 5-aminosalicylic acid may facilitate the transport of both 5-aminosalicylic acid and ursodeoxycholic acid to the colon and may be useful for the treatment of colonic inflammatory bowel diseases, ulcerative colitis and Crohn's disease.
Asunto(s)
Antiinflamatorios no Esteroideos/síntesis química , Antiinflamatorios no Esteroideos/metabolismo , Mucosa Intestinal/metabolismo , Mesalamina/síntesis química , Mesalamina/metabolismo , Ácido Ursodesoxicólico/análogos & derivados , Amidohidrolasas/metabolismo , Animales , Antiinflamatorios no Esteroideos/uso terapéutico , Transporte Biológico Activo , Clostridium perfringens/metabolismo , Pólipos del Colon/tratamiento farmacológico , Pólipos del Colon/metabolismo , Humanos , Enfermedades Inflamatorias del Intestino/tratamiento farmacológico , Enfermedades Inflamatorias del Intestino/metabolismo , Absorción Intestinal , Intestinos/microbiología , Mesalamina/uso terapéutico , Ratas , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/metabolismoRESUMEN
A novel calcium-chelating agent, N"-ursodeoxycholyldiethylenetriamine-N,N,N'-triacetic acid (UDCA-DTTA), was synthesized to study its ability to dissolve calcified gallstones. The chelating activity of the compound was demonstrated by dissolving calcium carbonate in vitro at a high dissolution rate. In the presence of the agent, sliced human gallstone with a composition of more than 50% calcium bilirubinate was thoroughly dissolved, indicating that calcium bilirubinate was dissolved from the gallstone. The ability to dissolve calcium was comparable to that of EDTA. However, the laminar structure of the sliced gallstone did not disappear in the presence of EDTA, whereas the structure disappeared in the presence of UDCA-DTTA. All these results indicate that UDCA-DTTA is an interesting compound as a parent substance for developing a prodrug for an oral or intravenous agent to dissolve calcium-containing gallstones.
Asunto(s)
Calcio , Quelantes/química , Colelitiasis/química , Ácido Ursodesoxicólico/análogos & derivados , Administración Oral , Amidohidrolasas/metabolismo , Animales , Bilis/química , Bilirrubina/química , Carbonato de Calcio/química , Quelantes/administración & dosificación , Quelantes/síntesis química , Quelantes/metabolismo , Estabilidad de Medicamentos , Ácido Edético/química , Humanos , Inyecciones Intravenosas , Mucosa Intestinal/enzimología , Hígado/enzimología , Masculino , Páncreas/enzimología , Pancreatina/metabolismo , Ratas , Ratas Sprague-Dawley , Ácido Ursodesoxicólico/administración & dosificación , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/química , Ácido Ursodesoxicólico/metabolismoRESUMEN
A newly synthesized conjugate of 7-hydroxy-3-O-phosphonocholic acid (ursodeoxycholic acid monophosphate) with 5-aminosalicylic acid (5-ASA-UDCA monophosphate) was investigated to determine its suitability for the evaluation of enteric bacteria. This compound, 5-ASA-UDCA monophosphate, was efficiently deconjugated by cholylglycine hydrolase to release 5-ASA, whereas it was completely resistant to deconjugation by pancreatic and intestinal mucosal enzymes. In everted gut sac experiments, 5-ASA-UDCA monophosphate was not actively absorbed from any part of the small intestine. In animal experiments, urinary excretions of N-acetyl-5-ASA (Ac-5ASA) were measured for 24 h following the oral administration of 20 mg of 5-ASA-UDCA monophosphate. Control rats excreted 276.3 +/- 89.0 micrograms (mean +/- S.E.) of Ac-5ASA, whereas the rats with intestinal bacterial overgrowth excreted more (1224.1 +/- 231.5 micrograms; p < 0.01). These basic studies indicate that this compound is likely to offer a simple method for the evaluation of microbial overgrowth without the use of radioisotopes or expensive, special apparatus.
Asunto(s)
Ácidos Aminosalicílicos , Bacterias/crecimiento & desarrollo , Intestinos/microbiología , Ácido Ursodesoxicólico/análogos & derivados , Ácidos Aminosalicílicos/síntesis química , Ácidos Aminosalicílicos/farmacocinética , Animales , Bilis/química , Humanos , Técnicas In Vitro , Mucosa Intestinal/metabolismo , Hígado/enzimología , Masculino , Pancreatina/metabolismo , Ratas , Ratas Wistar , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/farmacocinéticaRESUMEN
We studied the effect of 5 beta-cholane-3 alpha,7 beta,24-triol-24-sulfate (UDC-O-sulfate) on ileal active transport of cholyltaurine, on hepatic cholesterol 7 alpha-hydroxylase activity, on serum and liver cholesterol levels, on intestinal absorption of cholesterol, and on bile salt composition of gallbladder bile in hamsters. Experiments using the everted gut sacs show that UDC-O-sulfate inhibits the ileal active transport of cholyltaurine. In experiments feeding the diets with either UDC-O-sulfate, cholesterol, or both to hamsters, the addition of UDC-O-sulfate to the cholesterol-enriched diet reduced the elevation of serum and liver cholesterol levels caused by cholesterol feeding. Supplementation with UDC-O-sulfate to the standard diet was likely to reduce serum and liver cholesterol levels. Cholesterol 7 alpha-hydroxylase activity was higher in the two UDC-O-sulfate supplemented groups than in the two corresponding groups, the standard diet group and the cholesterol enriched diet group, respectively. Addition of UDC-O-sulfate to the standard and cholesterol diets did not change intestinal absorption of cholesterol. The change of the bile salt composition in hamsters fed UDC-O-sulfate may also suggest that the bile alcohol sulfate inhibits the intestinal absorption of endogenous bile salts. Hence the hypocholesterolemic activity of dietary UDC-O-sulfate is thought to be the effect of the partial interruption of the enterohepatic circulation of endogenous bile salts.
Asunto(s)
Anticolesterolemiantes/farmacología , Ácido Ursodesoxicólico/análogos & derivados , Animales , Anticolesterolemiantes/síntesis química , Colesterol/biosíntesis , Colesterol/sangre , Colesterol/metabolismo , Colesterol en la Dieta/farmacología , Cricetinae , Íleon/efectos de los fármacos , Íleon/metabolismo , Técnicas In Vitro , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Mesocricetus , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/farmacologíaRESUMEN
A new bile acid analogue, 3 alpha,7 alpha-dihydroxy-7 beta-methyl-24-nor-5 beta-cholan-23-oic acid (7-Me-norCDCA) was synthesized from the methyl ester of norursodeoxycholic acid, and its hepatic biotransformation was defined in the hamster. To synthesize 7-Me-norCDCA, the 3 alpha-hydroxyl group of methyl norursodeoxycholate was protected as the hemisuccinate, and the 7 beta-hydroxyl group was oxidized with CrO3 to form the 7-ketone. A Grigard reaction with methyl magnesium iodide followed by alkaline hydrolysis gave 7-Me-norCDCA (greater than 70% yield). The structure of the new compound was confirmed by proton magnetic resonance and mass spectrometry. After intraduodenal administration of the 14C-labeled compound into the anesthetized biliary fistula hamster, it was rapidly and efficiently secreted into the bile; 80% of radioactivity was recovered in 2 h. After intravenous infusion, the compound was efficiently extracted by the liver and secreted into the bile (greater than 75% in 3 h). Most (93%) of the biliary radioactivity was present in biotransformation products. The major biotransformation product (48.7 +/- 6.0%) was a new compound, assigned the structure of 3 alpha,5 beta,7 alpha- trihydroxy-7 beta-methyl-24-nor-5 beta-cholan-23-oic acid (5 beta-hydroxy-7- Me-norCDCA). In addition, conjugates of 7-Me-norCDCA with taurine (13.7 +/- 5.0%), sulfate (10.3 +/- 3.0%), or glucuronide (5.1 +/- 1.7%) were formed. 7-Me-norCDCA was strongly choleretic in the hamster; during its intravenous infusion, bile flow increased 2 to 3 times above the basal level, and the calculated choleretic activity of the compound (and its metabolic products) was much greater than that of many natural bile acids, indicating that the compound induced hypercholeresis. It is concluded that the biotransformation and physiological properties of 7-Me-norCDCA closely resemble those of norCDCA. Based on previous studies, the major biological effect of the 7-methyl group in 7-Me-norCDCA is to prevent its bacterial 7-dehydroxylation in the distal intestine.
Asunto(s)
Hígado/metabolismo , Ácido Ursodesoxicólico/análogos & derivados , Ácido Ursodesoxicólico/metabolismo , Animales , Fístula Biliar/metabolismo , Biotransformación , Colagogos y Coleréticos/síntesis química , Colagogos y Coleréticos/metabolismo , Colagogos y Coleréticos/farmacología , Cromatografía en Capa Delgada , Cricetinae , Cinética , Espectroscopía de Resonancia Magnética , Masculino , Espectrometría de Masas , Mesocricetus , Ácido Ursodesoxicólico/síntesis química , Ácido Ursodesoxicólico/farmacologíaRESUMEN
In order to study the glycosidic conjugation of chenodeoxycholic, hyodeoxycholic, and ursodeoxycholic acids in patients with cholestasis after oral administration of pharmacological amounts of the respective bile acids avoiding the application of radioactive tracers we synthesized [24-13C]chenodeoxycholic, [24-13C]hyodeoxycholic, and [24-13C]ursodeoxycholic acids. The reaction intermediates of the bile acid syntheses were characterized by infrared spectroscopy. Purity was confirmed using thin-layer chromatography as well as gas chromatography-mass spectrometry. The 13C atom excess of approximately 90% of the synthesized bile acids was the same as the 13C atom excess of the sodium [13C]cyanide used for the labeling reaction confirming the successful synthesis. After oral administration of 0.5 g of [24-13C]ursodeoxycholic acid to a healthy volunteer, 13C label was detected in the nonamidated and glycine- or taurine conjugated glucosides and the N-acetylglucosaminide of ursodeoxycholic acid in urine. This establishes ursodeoxycholic acid as the first bile acid so far known to undergo both of the recently described glycosidic conjugation reactions in humans.