64284-64-6Relevant academic research and scientific papers
The investigational drug VT-1129 is a highly potent inhibitor of Cryptococcus species CYP51 but only weakly inhibits the human enzyme
Warrilow, Andrew G. S.,Parker, Josie E.,Price, Claire L.,Nes, W. David,Garvey, Edward P.,Hoekstra, William J.,Schotzinger, Robert J.,Kelly, Diane E.,Kelly, Steven L.
, p. 4530 - 4538 (2016)
Cryptococcosis is a life-threatening disease often associated with HIV infection. Three Cryptococcus species CYP51 enzymes were purified and catalyzed the 14α-demethylation of lanosterol, eburicol, and obtusifoliol. The investigational agent VT-1129 bound tightly to all three CYP51 proteins (dissociation constant [Kd] range, 14 to 25 nM) with affinities similar to those of fluconazole, voriconazole, itraconazole, clotrimazole, and ketoconazole (Kd range, 4 to 52 nM), whereas VT-1129 bound weakly to human CYP51 (Kd, 4.53 μM). VT-1129 was as effective as conventional triazole antifungal drugs at inhibiting cryptococcal CYP51 activity (50% inhibitory concentration [IC50] range, 0.14 to 0.20 μM), while it only weakly inhibited human CYP51 activity (IC50, ～600 μM). Furthermore, VT-1129 weakly inhibited human CYP2C9, CYP2C19, and CYP3A4, suggesting a low drug-drug interaction potential. Finally, the cellular mode of action for VT-1129 was confirmed to be CYP51 inhibition, resulting in the depletion of ergosterol and ergosta-7-enol and the accumulation of eburicol, obtusifolione, and lanosterol/obtusifoliol in the cell membranes.
An alternative synthesis of 4,4-dimethyl-5α-cholesta-8,14,24-trien-3β-ol, an intermediate in sterol biosynthesis and a reported activator of meiosis and of nuclear orphan receptor LXR
Ruan,Wilson,Schroepfer Jr.
, p. 233 - 236 (1998)
4,4-Dimethyl-5α-cholesta-8,14,24-trien-3β-ol, a sterol of current biological interest, has been synthesized in six steps from 3β-acetoxy-4,4-dimethyl-5α-cholest-8(14)-en-15-one.
Functional importance for developmental regulation of sterol biosynthesis in Acanthamoeba castellanii
Zhou, Wenxu,Warrilow, Andrew G.S.,Thomas, Crista D.,Ramos, Emilio,Parker, Josie E.,Price, Claire L.,Vanderloop, Boden H.,Fisher, Paxtyn M.,Loftis, Michael D.,Kelly, Diane E.,Kelly, Steven L.,Nes, W. David
, p. 1164 - 1178 (2018)
The sterol metabolome of Acanthamoeba castellanii (Ac) yielded 25 sterols. Substrate screening of cloned AcCYP51 revealed obtusifoliol as the natural substrate which converts to ?8,14-sterol (2H3-methyl]methionine incubation to intact cultures showing C28-ergosterol incorporates 2-2H atoms and C29-7-dehydroporiferasterol incorporates 5 2H-atoms, the natural distribution of sterols, CYP51 and previously published sterol methyltransferase (SMT) data indicate separate ?24(28)- and ?25(27)-olefin pathways to C28- and C29-sterol products from the protosterol cycloartenol. In cell-based culture, we observed a marked change in sterol compositions during the growth and encystment phases monitored microscopically and by trypan blue staining; trophozoites possess C28/C29-?5,7-sterols, viable encysted cells (mature cyst) possess mostly C29-?5-sterol and non-viable encysted cells possess C28/C29-?5,7-sterols that turnover variably from stress to 6-methyl aromatic sterols associated with changed membrane fluidity affording lysis. An incompatible fit of steroidal aromatics in membranes was confirmed using the yeast sterol auxotroph GL7. Only viable cysts, including those treated with inhibitor, can excyst into trophozoites. 25-Azacycloartanol or voriconazole that target SMT and CYP51, respectively, are potent enzyme inhibitors in the nanomolar range against the cloned enzymes and amoeba cells. At minimum amoebicidal concentration of inhibitor amoeboid cells rapidly convert to encysted cells unable to excyst. The correlation between stage-specific sterol compositions and the physiological effects of ergosterol biosynthesis inhibitors suggests that amoeba fitness is controlled mainly by developmentally-regulated changes in the phytosterol B-ring; paired interference in the ?5,7-sterol biosynthesis (to ?5,7) - metabolism (to ?5 or 6-methyl aromatic) congruence during cell proliferation and encystment could be a source of therapeutic intervention for Acanthamoeba infections.
Characterization of a novel CYP51 from Rhodococcus triatomae and its NADH-ferredoxin reductase-coupled application in lanosterol 14α-demethylation
Ke, Xia,Ding, Guan-Jun,Ma, Bin-Xiang,Liu, Zhi-Qiang,Zhang, Jin-Feng,Zheng, Yu-Guo
, p. 59 - 68 (2017)
Reconstitution of a selective demethylation system for lanosterol is desperately needed for more efficient synthesis of steroidal drugs. Sterol 14α-demethylase cytochrome P450 (CYP51) has been confirmed to catalyze sterol 14α-demethylation, an essential reaction in sterol biosynthesis. Herein, a putative CYP51 gene (RtCYP51) was mined from the complete genome sequence of Rhodococcus triatomae BKS 15-14. Its amino acid sequence showed 25–68% identity to other sterol 14α-demethylases, and contained a novel alanine-rich sequence at the C-terminus. Heterologous expression of the RtCYP51 gene in Escherichia coli (E. coli) yielded a ～54 kDa recombination protein that exhibited a typical reduced CO-difference spectrum and a dissociation constant (Kd) of 2.93 μM for lanosterol. Furthermore, three exogenous electron donor systems, including Fdx-FdR (Acinetobacter sp.OC4 ferredoxin and ferredoxin reductase), Fld-FdR2 (E. coli flavodoxin and flavodoxin reductase) and NfFdR (Nocardia farcinica iron-sulfur containing NADPH-P450 reductase) were selected for coupling the electron-transfer from the coenzyme to RtCYP51. Fdx-FdR was found to be the most efficient electron donor and was also confirmed to support the lanosterol demethylation activity of RtCYP51 in vitro. Under the optimum molar ratio of RtCYP51/FdR/Fdx (1:2:10), RtCYP51 exhibited a relatively high turnover number of 0.63 min?1 (nmol metabolized lanosterol/min/nmol RtCYP51), compared with known bacterial CYP51s.
Synthesis of FF-MAS from lithocholic acid
Murray, Anthony,Gronvald, Frederik C.,Nielsen, Jane K.,Faarup, Peter
, p. 1067 - 1068 (2000)
An effective synthesis of 4,4 dimethyl-cholest-8,14,24-trien-3β-ol (FF-MAS) from lithocholic acid is described, utilising a double oxidation and regioselective Wittig reaction as key steps. (C) 2000 Elsevier Science Ltd. All rights reserved.
Heterologous expression and characterization of the sterol 14α-demethylase CYP51F1 from Candida albicans
Park, Hyoung-Goo,Lee, Im-Soon,Chun, Young-Jin,Yun, Chul-Ho,Johnston, Jonathan B.,Montellano, Paul R. Ortiz De,Kim, Donghak
experimental part, p. 9 - 15 (2012/02/15)
Lanosterol 14α-demethylase (CYP51F1) from Candida albicans is known to be an essential enzyme in fungal sterol biosynthesis. Wild-type CYP51F1 and several of its mutants were heterologously expressed in Escherichia coli, purified, and characterized. It exhibited a typical reduced CO-difference spectrum with a maximum at 446 nm. Reconstitution of CYP51F1 with NADPH-P450 reductase gave a system that successfully converted lanosterol to its demethylated product. Titration of the purified enzyme with lanosterol produced a typical type I spectral change with Kd = 6.7 μM. The azole antifungal agents econazole, fluconazole, ketoconazole, and itraconazole bound tightly to CYP51F1 with Kd values between 0.06 and 0.42 μM. The CYP51F1 mutations F105L, D116E, Y132H, and R467K frequently identified in clinical isolates were examined to determine their effect on azole drug binding affinity. The azole Kd values of the purified F105L, D116E, and R467K mutants were little altered. A homology model of C. albicans CYP51F1 suggested that Tyr132 in the BC loop is located close to the heme in the active site, providing a rationale for the modified heme environment caused by the Y132H substitution. Taken together, functional expression and characterization of CYP51F1 provide a starting basis for the design of agents effective against C. albicans infections.
Process for the production of 4,4-dimethyl-5α-cholesta-8,14,24-trien-3β-01 and intermediate products in process (I)
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Page column 11, (2010/02/05)
The subject of this invention is a new process with variant embodiments for the production of compound 1.
Two syntheses of FF-MAS
Blume, Thorsten,Guttzeit, Marc,Kuhnke, Joachim,Zorn, Ludwig
, p. 1837 - 1839 (2007/10/03)
(Matrix presented) Follicular fluid-meiosis activating sterol (FF-MAS) has been shown to be an efficient inducer of meiotic maturation. It can potentially be used for improvements of in vitro fertilization techniques. Two short synthesis of FF-MAS are pre
Synthesis of zymosterol, fecosterol, and related biosynthetic sterol intermediates
Dolle,Schmidt,Erhard,Kruse
, p. 278 - 284 (2007/10/02)
The first syntheses of sterol biosynthetic intermediates zymosterol(4), 4,4-dimethylzymosterol(5), cholesta-8,14,24-trien-3β-ol(6), the 4,4-dimethyl analogue 7, and fecosterol (8) are described in detail. Multigram quantities of key intermediates 16 and 17 were efficiently prepared from known enones 20 and 21 (eight steps, 35% overall yield). Novel entry into Δ8-sterols was achieved through regiospecific hydroboration/deoxygenation of the 8,14-diene systems. Sterols containing Δ24- or Δ(24(28))-olefins were obtained from C24-hydroxy intermediates either via dehydration using bis[α,α-bis(trifluoromethyl)benzenemethanolato]diphenylsulfur in CH2Cl2 or via Swern oxidation/Wittig olefination, respectively. In this way, 16 and 17 were converted to the desired Δ8,24-, Δ8,14,24-, and Δ(8,24(28))-sterols with high regiocontrol.
