- Engineering Regioselectivity of a P450 Monooxygenase Enables the Synthesis of Ursodeoxycholic Acid via 7β-Hydroxylation of Lithocholic Acid
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We engineered the cytochrome P450 monooxygenase CYP107D1 (OleP) from Streptomyces antibioticus for the stereo- and regioselective 7β-hydroxylation of lithocholic acid (LCA) to yield ursodeoxycholic acid (UDCA). OleP was previously shown to hydroxylate testosterone at the 7β-position but LCA is exclusively hydroxylated at the 6β-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.
- Grobe, Sascha,Badenhorst, Christoffel P. S.,Bayer, Thomas,Hamnevik, Emil,Wu, Shuke,Grathwol, Christoph W.,Link, Andreas,Koban, Sven,Brundiek, Henrike,Gro?johann, Beatrice,Bornscheuer, Uwe T.
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supporting information
p. 753 - 757
(2020/12/01)
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- Chenodeoxycholic acid derivative or pharmaceutically acceptable salt thereof, and preparation method and applications thereof
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The invention discloses a chenodeoxycholic acid derivative with a structure as shown in general formula I or a medicinal salt thereof, and a preparation method and application of the chenodeoxycholicacid derivative. The chenodeoxycholic acid derivative can up-regulate the transcription levels of FXR mRNA and SHP mRNA, can obviously activate FXR, and can be used for preparing drugs for treating orpreventing hyperlipidemia, atherosclerosis, non-alcoholic steatohepatitis, type II diabetes mellitus and other diseases related to blood fat.
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Paragraph 0042; 0043; 0044
(2020/03/12)
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- Chemical Synthesis of Rare Natural Bile Acids: 11α-Hydroxy Derivatives of Lithocholic and Chenodeoxycholic Acids
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A method for the preparation of 11α-hydroxy derivatives of lithocholic and chenodeoxycholic acids, recently discovered to be natural bile acids, is described. The principal reactions involved were (1) elimination of the 12α-mesyloxy group of the methyl esters of 3α-acetate-12α-mesylate and 3α,7α-diacetate-12α-mesylate derivatives of deoxycholic acid and cholic acid with potassium acetate/hexamethylphosphoramide; (2) simultaneous reduction/hydrolysis of the resulting △11-3α-acetoxy and △11-3α,7α-diacetoxy methyl esters with lithium aluminum hydride; (3) stereoselective 11α-hydroxylation of the △11-3α,24-diol and △11-3α,7α,24-triol intermediates with B2H6/tetrahydrofuran (THF); and (4) selective oxidation at C-24 of the resulting 3α,11α,24-triol and 3α,7α,11α,24-tetrol to the corresponding C-24 carboxylic acids with NaClO2 catalyzed by 2,2,6,6-tetramethylpiperidine 1-oxyl free radical (TEMPO) and NaClO. In summary, 3α,11α-dihydroxy-5β-cholan-24-oic acid and 3α,7α,11α-trihydroxy-5β-cholan-24-oic acid have been synthesized and their nuclear magnetic resonance (NMR) spectra characterized. These compounds are now available as reference standards to be used in biliary bile acid analysis.
- Namegawa, Kazunari,Iida, Kyoko,Omura, Kaoru,Ogawa, Shoujiro,Hofmann, Alan F.,Iida, Takashi
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p. 403 - 411
(2018/06/08)
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- Chenodeoxycholic acid derivatives, preparation method thereof and medical application thereof
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The invention relates to the field of medicinal chemistry, relates to chenodeoxycholic acid derivatives, a preparation method thereof and a medical application thereof, in particular to a kind of chenodeoxycholic acid derivatives with a general formula of (I), a preparation method thereof, a pharmaceutical composition comprising the compounds and medical application thereof, especially used as drugs for preventing or treating hyperlipidaemia, type II diabetes, atherosis and non-alcoholic steatohepatitis. The formula is shown in the description.
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Paragraph 0042; 0043; 0044
(2018/06/26)
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- Lipid accumulation inhibitory activities of novel isoxazole-based chenodeoxycholic acids: Design, synthesis and preliminary mechanism study
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In continuation of our drug discovery program on hyperlipidemia, a series of novel isoxazole-chenodeoxycholic acid hybrids were designed, synthesized and evaluated for their lipid-lowering effects. Preliminary screening of all the synthesized compounds was done by using a 3T3-L1 adipocyte model, in which the most active compound 16b could significantly reduce the lipid accumulation up to 30.5% at a nontoxic concentration 10 μM. Further mechanism studies revealed that 16b blocked lipid accumulation via activating FXR-SHP signaling pathway, efficiently down-regulated the expression of key lipogenesis regulator SREBP-1c.
- Qiu, Rongmao,Luo, Guoshun,Li, Xinyu,Zheng, Fan,Li, Haolin,Zhang, Jin,You, Qidong,Xiang, Hua
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p. 2879 - 2884
(2018/07/25)
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- Chemical synthesis of uncommon natural bile acids: The 9α-hydroxy derivatives of chenodeoxycholic and lithocholic acids
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The chemical synthesis of the 9α-hydroxy derivatives of chenodeoxycholic and lithocholic acids is reported. For initiating the synthesis of the 9α-hydroxy derivative of chenodeoxycholic acid, cholic acid was used; for the synthesis of the 9α-hydroxy derivative of lithocholic acid, deoxycholic acid was used. The principal reactions involved were (1) decarbonylation of conjugated 12-oxo-Δ9(11)-derivatives using in situ generated monochloroalane (AlH2Cl) prepared from LiAlH4 and AlCl3, (2) epoxidation of the deoxygenated Δ9(11)-enes using m-chloroperbenzoic acid catalyzed by 4,4'-thiobis-(6-tert-butyl-3-methylphenol), (3) subsequent Markovnikov 9a-hydroxylation of the Δ9(11)-enes with AlH2Cl, and (4) selective oxidation of the primary hydroxyl group at C-24 in the resulting 3α,9α,24-triol and 3α,7α,9α,24-tetrol to the corresponding C-24 carboxylic acids using sodium chlorite (NaClO2) in the presence of a catalytic amount of 2,2,6,6-tetramethylpiperidine 1-oxyl free radical (TEMPO) and sodium hypochlorite (NaOCl). The 1H- and 13C-NMR spectra are reported. The 3α,7α,9α-trihydroxy-5β-cholan-24-oic acid has been reported to be present in the bile of the Asian bear, and its 7-deoxy derivative is likely to be a bacterial metabolite. These bile acids are now available as authentic reference standards, permitting their identification in vertebrate bile acids.
- Iida, Takashi,Namegawa, Kazunari,Nakane, Naoya,Iida, Kyoko,Hofmann, Alan Frederick,Omura, Kaoru
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p. 1397 - 1402
(2016/10/03)
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- 11 -SUBSTITUTED BILE ACID DERIVATIVES, PROCESS FOR THE PREPARATION THEREOF AND USE OF THESE COMPOUNDS AS MEDICAMENTS
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The present invention discloses a novel bile acid derivatives having substituted nitrogen functionality at C-11 and process for synthesis thereof. These C-11 substituted bile acid derivatives shows anticancer and antimycobacterial activity.
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Paragraph 0053
(2015/06/25)
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- Synthesis, NMR Characterization and Crystal Structure of Methyl 3α,7α-Dihydroxy-12-oxo-5β-cholanate
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The crystal structure and NMR characterization of methyl 3α,7α-dihydroxy-12-oxo-5β-cholanate are described. The title compound which was obtained from methyl cholanate in a 3-step synthetic sequence that does not alter the starting chirality, crystallizes in the monoclinic system with P 21 space group. While despite the substitution pattern rings A, B and C adopt chair conformations, the 5 membered D ring, that bears the side chain attached to C-17, shows a twisted conformation on C-13-C-14. In the crystal array, classical hydrogen bond interactions O-H···H and intermolecular contacts C-H···O of hydrogen bond type are observed. Graphical Abstract: The crystal structure and NMR characterization of Methyl α,7α-dihydroxy-12-oxo-5β-cholanate are described. [Figure not available: see fulltext.]
- Tinajero-Delgado, Vernica,Romero-vila, Margarita,Flores-lamo, Marcos,Arteaga, Martn A. Iglesias
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p. 487 - 492
(2015/02/19)
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- PROCESS FOR PREPARING HIGH PURITY URSODEOXYCHOLIC ACID
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The present invention describes a process for the synthesis of ursodeoxycholic acid wherein the purification of the crude ursodeoxycholic acid (containing approximately 13-15% of chenodeoxycholic acid impurity) takes place first passing through a salification with imidazole and a subsequent purification via "methyl ester", which allows a finished product with an extremely low content of known "cheno and "litho" impurities to be obtained. The present invention also describes the recovery steps of cholic acid and 3α-hydroxy-7-ketocholanic acid from the mother liquors of process intermediates.
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Page/Page column 4; 7; 8
(2014/02/16)
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- Stereoselective synthesis of pentacyclic steroids functionalized at C-11
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We set out to describe an efficient and versatile method for preparing pentacyclic steroids diversely substituted at C-11 from cholic acid, via a stereoselective epoxidation and the epoxide opening as the key steps. The characteristic 1H and 13C NMR spectroscopic features of the synthesized compounds are reported.
- Ibrahim-Ouali, Malika,Romero, Eugenie,Hamze, Khalil
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p. 1092 - 1100
(2012/11/13)
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- A ratiometric TICT-type dual fluorescent sensor for an amino acid
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A novel ratiometric fluorescent sensor 1 having the 4-(N,N-dimethylamino) benzoate (DMAB) group as a twisted intramolecular charge transfer (TICT) dual fluorescence site and the guanidinium group as a binding site for an amino acid at the 12- and 3-positions of cholic acid, respectively, was designed and synthesized. The sensor 1 showed characteristic dual fluorescence and it was shown that the guanidinium group in the proximity of the DMAB group influenced largely the TICT formation process in 1. Liquid/liquid extraction experiments demonstrated that the intensity ratio of the TICT fluorescence to the locally-excited (LE) fluorescence (ITICT/ILE) observed from the organic layer decreased with an increase in the concentration of N-acetyl-d-phenylalanine (AcPhe) in the water phase. Such fluorescence titration experiments afforded the apparent binding constant between 1 and AcPhe to be 7.0 × 105 M-1. Temperature dependence of the fluorescence spectrum of 1 in the absence and presence of AcPhe indicated that the primary origin of the change in the ITICT/ILE value upon recognition of the amino acid was the increase in the activation energy of the TICT formation process in 1. the Owner Societies.
- Ito, Akitaka,Ishizaka, Shoji,Kitamura, Noboru
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experimental part
p. 6641 - 6649
(2011/09/14)
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- New synthesis of pentacyclic steroids by stereoselective epoxide ring opening
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A stereoselective synthesis of pentacyclic steroids has been achieved. Starting from commercially available cholic acid 1, followed by asymmetric epoxidation and by stereoselective epoxide ring opening, employing nucleophilic species, the corresponding products were afforded in good yields. The compounds were being evaluated for their biological activity.
- Ibrahim-Ouali, Malika,Khalil, Hamze
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scheme or table
p. 6948 - 6950
(2011/03/18)
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- Molecular association via halogen bonding and other weak interactions in the crystal structures of 11-bromo-12-oxo-5β-cholan derivatives
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Methyl 3α,7α-diacetoxy-12-oxo-5β-cholan-24-oate 2, methyl 11α-bromo-3α,7α-diacetoxy-12-oxo-5β-cholan-24-oate 3, methyl 11β-bromo-3α,7α-diacetoxy-12-oxo-5β-cholan-24-oate 4 and methyl 11,11-dibromo-3α,7α-diacetoxy-12-oxo-5β-cholan-24-oate 5 were synthesized. The crystal structures of these molecules were resolved to study the effect of bulky bromine atom in the steroid skeleton of cholic acid with different stereo-chemical orientations at C-11 on the two-dimensional arrangement of molecules and solid-state properties. All the molecules associate only via weak intermolecular interactions in their crystal structures, notable one being the Halogen Bonded assembly (C-Br...O) in 5.
- Salunke, Deepak B.,Hazra, Braja G.,Gonnade, Rajesh G.,Pore, Vandana S.,Bhadbhade, Mohan M.
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experimental part
p. 246 - 253
(2009/03/12)
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- Amino functionalized novel cholic acid derivatives induce HIV-1 replication and syncytia formation in T cells
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Synthesis of C-11 azido/amino functionalized cholic acid derivatives has been achieved in excellent yields. Contrary to the previous prediction of analogous compounds to be HIV-1 protease inhibitors, in the present study these novel cholic acid derivatives induced host cell fusion during the progress of HIV-1 infection and produced multinucleated giant cells. This is the first report of syncytia induction and enhancement of viral replication in HIV-1 infected T cells by cholic acid derivatives.
- Salunke, Deepak B.,Ravi, Dyavar S.,Pore, Vandana S.,Mitra, Debashis,Hazra, Braja G.
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p. 2652 - 2655
(2007/10/03)
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- Chemical modifications of bile acids under high-intensity ultrasound or microwave irradiation
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High-intensity ultrasound (HIU) and microwave (MW) irradiation, having emerged as effective promoters of organic reactions, were exploited for the synthesis of bile acids derivatives. Esterification, amidation, hydrolysis, oxidation, and reduction were investigated. Compared to conventional methods, both techniques proved much more efficient, increasing product yields and dramatically cutting down reaction times. Scaled-up studies are now under way.
- Cravotto, Giancarlo,Boffa, Luisa,Turello, Marta,Parenti, Massimo,Barge, Alessandro
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- Liver-selective glucocorticoid antagonists: A novel treatment for type 2 diabetes
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Hepatic blockade of glucocorticoid receptors (GR) suppresses glucose production and thus decreases circulating glucose levels, but systemic glucocorticoid antagonism can produce adrenal insufficiency and other undesirable side effects. These hepatic and systemic responses might be dissected, leading to liver-selective pharmacology, when a GR antagonist is linked to a bile acid in an appropriate manner. Bile acid conjugation can be accomplished with a minimal loss of binding affinity for GR. The resultant conjugates remain potent in cell-based functional assays. A novel in vivo assay has been developed to simultaneously evaluate both hepatic and systemic GR blockade; this assay has been used to optimize the nature and site of the linker functionality, as well as the choice of the GR antagonist and the bile acid. This optimization led to the identification of A-348441, which reduces glucose levels and improves lipid profiles in an animal model of diabetes.
- Von Geldern, Thomas W.,Tu, Noah,Kym, Philip R.,Link, James T.,Jae, Hwan-Soo,Lai, Chunqiu,Apelqvist, Theresa,Rhonnstad, Patrik,Hagberg, Lars,Koehler, Konrad,Grynfarb, Marlena,Goos-Nilsson, Annika,Sandberg, Johnny,Osterlund, Marie,Barkhem, Tomas,H?glund, Marie,Wang, Jiahong,Fung, Steven,Wilcox, Denise,Nguyen, Phong,Jakob, Clarissa,Hutchins, Charles,F?rnegf?rdh, Mathias,Kauppi, Bj?rn,?hman, Lars,Jacobson, Peer B.
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p. 4213 - 4230
(2007/10/03)
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- Cyclocholates with 12-Oxo and 7,12-Oxo Groups
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Syntheses of bile acid cyclooligomers with 12- and 7,12-oxo groups (6a-d, 7a-c, 8a-b) by the Yamaguchi method are described. Cyclotrimerization is the principal reaction route for these cholic acid systems. Conversion of 7- and 12-hydroxy groups in cholic acid (la-b) to oxo groups (4a-c, 5a-c), followed by macrocyclization (6a-d, 7a-c, 8a-b) and selective reduction of the oxo groups back to hydroxy ones without cleaving the 24-carboxylic ester linkages (11) constitutes a new strategy in the synthesis of cyclocholates having unprotected hydroxy groups.
- Gao, Hongwu,Dias, Jerry Ray
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p. 719 - 724
(2007/10/03)
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- Substituted Methyl 5β-Cholan-24-oates. I - 17O NMR Spectral Characterization
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Methyl esters of four common bile acids, 3α-hydroxy-5β-cholan-24-oic (lithocholic) acid, 3α,7α-dihydroxy-5β-cholan-24-oic (chenodeoxycholic) acid, 3α,12α-dihydroxy-5β-cholan-24-oic (deoxycholic) acid and 3α,7α,12α-trihydroxy-5β-cholan-24-oic (cholic) acid, and 14 acetylated, trifluoroacetylated, mesylated and oxo derivatives of methyl 5β-cholan-24-oates were prepared and their 17O NMR spectra recorded.In spite of their relatively high molecular masses and the rigid molecular structure of the steroid skeleton, most of the oxygens included in these structures gave well resolved 17O NMR resonance lines at natural abundance in 0.25-0.5 M acetonitrile solutions at 75 deg C.In agreement with the present 17O NMR results, molecular mechanics calculations revealed that a hydroxy substituent located at the 3α-position clearly differs from the hydroxyls at the 7α- and 12α-positions.This is due to the fact that the 3α-hydroxyl possesing only two γ-carbons at antiperiplanar positions is less shielded than the other hydroxyls influenced also by the shielding effects of γ-gauche carbons.The spectral deconvolution of the overlapping signals of the 7α- and 12α-hydroxyls is based on a computer-aided method or on chemical substitutions.The oxo groups located at the longitudinal (3-oxo) vs. transversal (7- and 12-oxo) axes of the steroid framework show very different quadrupolar relaxation properties and 17O NMR linewidths owing to the strong anisotropy of overall molecular motion.In contrast, the 17O NMR linewidths of all 3α-, 7α- and 12α-hydroxyls are very similar and clearly smaller than those of the corresponding oxo groups, revealing that their quadrupolar relaxtion is merely determined by their internal rotation rather than by the overall molecular motion. - Keywords: NMR 17O NMR 17O chemical shifts 17O relaxation Methyl 5β-cholan-24-oates Methyl esters of bile acids
- Kolehmainen, Erkki,Kaartinen, Mari,Kauppinen, Reijo,Kotoneva, Jari,Lappalainen, Kari,et al.
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p. 441 - 445
(2007/10/02)
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- REDUCTION OF 12-OXO DERIVATIVES OF SOME BILE ACIDS
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Methyl ester of 3α-acetoxy-12-oxo-9 (11)-cholenic acid and some other analogous compounds were reduced to the corresponding 9(11)-unsaturated 12-hydroxy derivatives by the action of sodium borohydride in the presence of cerium(III) chloride.A part of our work, viz. preparation of steroid hormones from bile acids, was the reduction of 9(11)-unsaturated 12-oxo derivatives of some bile acids, with the view of obtaining the corresponding 9(11)-unsaturated 12-hydroxy derivatives.Hitherto, the only safe route has been hydrogenation of starting 9(11)-unsaturated 12-oxo derivatives on platinum at elevated pressures, which, preserving the allyl double bond, yields a mixture of the two isomeric 12-hydroxy derivatives.
- Mickova, Ruzena
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p. 1239 - 1243
(2007/10/02)
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- Introducing Δ11 unsaturation into steroid compounds
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A process for the introduction of Δ11 unsaturation into steroid compounds having a C-12 sulfonate ester group is described. Dehydrosulfonation is carried out by reacting the sulfonate with a hexaalkylphosphoric triamide. The process is particularly useful with steroid compounds that also contain a blocked C-7 hydroxy group. The process yields the 11-enate in preference to the 6,11-dienate. The yield of the process can be increased by carrying out the reaction in the presence of a weak base.
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