Assay of 3-hydroxy-3-methylglutaryl CoA reductase activity using anionic-exchange column chromatography

Ong, K.K.; Khor, H. T.; Tan, D.T.S.

doi:10.1016/0003-2697(91)90455-3

Cited by 11 publications

(4 citation statements)

References 8 publications

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“…All analogues were evaluated for inhibitory potency in a HMG-CoA reductase enzyme inhibition assay. 22,23 Additionally, the ability of analogues to block cholesterol synthesis in both rat hepatocyte and myocyte cell lines was evaluated; moreover, comparison of these two values was utilized as a measurement of hepatoselectivity. 22,23 Table 1 shows the molecular structure, biological activity, and ITC results for the binding interaction of inhibitors with HMGR at 30 °C for a set of structurally diverse inhibitors clustered into six template subtypes (in addition to the substrate HMG-CoA and benchmark rosuvastatin).…”

Section: Resultsmentioning

confidence: 99%

“…Starting with several structurally diverse templates, compounds were synthesized and binding affinity to HMGR was determined using ITC. All analogues were evaluated for inhibitory potency in a HMG-CoA reductase enzyme inhibition assay. , Additionally, the ability of analogues to block cholesterol synthesis in both rat hepatocyte and myocyte cell lines was evaluated; moreover, comparison of these two values was utilized as a measurement of hepatoselectivity. , Table shows the molecular structure, biological activity, and ITC results for the binding interaction of inhibitors with HMGR at 30 °C for a set of structurally diverse inhibitors clustered into six template subtypes (in addition to the substrate HMG-CoA and benchmark rosuvastatin). A high resolution costructure for each of the inhibitors in Table was also solved with HMGR and representative structures from each class deposited in the Protein Data Bank.…”

Section: Resultsmentioning

confidence: 99%

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Thermodynamic and Structure Guided Design of Statin Based Inhibitors of 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase

Sarver¹,

Bills²,

Bolton³

et al. 2008

J. Med. Chem.

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Clinical studies have demonstrated that statins, 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) inhibitors, are effective at lowering mortality levels associated with cardiovascular disease; however, 2-7% of patients may experience statin-induced myalgia that limits compliance with a treatment regimen. High resolution crystal structures, thermodynamic binding parameters, and biochemical data were used to design statin inhibitors with improved HMGR affinity and therapeutic index relative to statin-induced myalgia. These studies facilitated the identification of imidazole 1 as a potent (IC 50 = 7.9 nM) inhibitor with excellent hepatoselectivity (>1000-fold) and good in vivo efficacy. The binding of 1 to HMGR was found to be enthalpically driven with a Delta H of -17.7 kcal/M. Additionally, a second novel series of bicyclic pyrrole-based inhibitors was identified that induced order in a protein flap of HMGR. Similar ordering was detected in a substrate complex, but has not been reported in previous statin inhibitor complexes with HMGR.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Thermodynamic and Structure Guided Design of Statin Based Inhibitors of 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase

Sarver¹,

Bills²,

Bolton³

et al. 2008

J. Med. Chem.

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“…Flash chromatography (ethyl acetate) gave 17 (240 mg), 18 (270 mg), and a mixture of 17 and 18 (240 mg « 1:1), total yield of diols 75%. 17 (unpolar isomer, Rf = 0.40, ethyl acetate/methanol (19). 17 (200 mg, 0.40 mmol), 2,2-dimethoxypropane (0.1 mL), acetone (5 mL), and p-toluenesulfonic acid (5 mg) were stirred at room temperature for 1 h. Triethylamine (0.05 mL) was added and the mixture evaporated.…”

Section: Methodsmentioning

confidence: 99%

“…Enzymatic Activity of HMG-CoA Reductase. 19 For investigations of the effects of HMG-CoA reductase inhibitors on the enzymatic activity of HMG-CoA reductase, microsomes from rat liver were prepared as follows. Male Wistar rats (weighing 200-220 g) were fed a standard diet (Altromin) containing 2% cholestyramine for 10 days under reversed light cycle.…”

Section: -(R)-[2-[3a-hydroxy-12a-(2-(s)mentioning

confidence: 99%

Synthesis and Biological Activity of Bile Acid-Derived HMG-CoA Reductase Inhibitors. The Role of 21-Methyl in Recognition of HMG-CoA Reductase and the Ileal Bile Acid Transport System

Wess¹,

Kramer²,

Han³

et al. 1994

J. Med. Chem.

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To increase hepatoselectivity of HMG-CoA reductase inhibitors by using the specific bile acid transport systems, deoxycholic acid-derived inhibitors 9 and 11 have been synthesized, on the basis of the concept of combining in one molecule structural requirements for specific inhibition of the HMG-CoA reductase and specific recognition by the ileal bile acid transport system. The 1-methyl-3-carboxylpropyl subunit of deoxycholic acid was replaced by the 3,5-dihydroxyheptanoic acid lactone of lovastatin, and position 12-OH was esterified with 2-methylbutyric acid. Compounds 9 and 11 were evaluated for their inhibitory activity on rat liver HMG-CoA reductase, cholesterol biosynthesis in HEP G2 cells, and [3H]taurocholate uptake in rabbit brush border membrane vesicles and compared with methyl derivatives 8 and 10. The steroidal 21-CH3 group affects both activity on HMG-CoA reductase and recognition by the ileal bile acid transport system.

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Simultaneous evaluation of HMG‐CoA reductase and cholesterol 7α‐hydroxylase activities by electrospray tandem MS

Ndong-Akoume

Mignault

Perwaiz

et al. 2002

Lipids

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Simultaneous evaluation of HMG-CoA reductase and cholesterol 7alpha-hydroxylase activities by electrospray tandem mass spectrometry (ES-MS-MS) was performed. The assay was based on the measurement of mevalonolactone (MVL) and 7alpha-hydroxycholesterol (7alpha-OHC) produced by the incubation of HMG-CoA with hepatic microsomes in the presence of NADPH and glucose-6-phosphate dehydrogenase. Following extraction and purification using a cyanopropyl cartridge, MVL and 7alpha-OHC were analyzed, without derivatization, by ES-MS-MS. The analysis was achieved in 5 min. Calibration curves were made for MVL and 7alpha-OHC, and were linear from 0 to 100 microg. The recovery was >97%. The procedure was validated under similar calibration and recovery experiments, by measuring the above mentioned products as dimethylethylsilyl ether derivatives using the classical technique of GC-MS. Data obtained by ES-MS-MS and GC-MS showed a good correlation, with no significant differences. ES-MS-MS is a simple and reliable method for the evaluation of HMG-CoA reductase and cholesterol 7alpha-hydroxylase activities in liver microsomal preparations.

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Assay of 3-hydroxy-3-methylglutaryl CoA reductase activity using anionic-exchange column chromatography

Cited by 11 publications

References 8 publications

Thermodynamic and Structure Guided Design of Statin Based Inhibitors of 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase

Thermodynamic and Structure Guided Design of Statin Based Inhibitors of 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase

Synthesis and Biological Activity of Bile Acid-Derived HMG-CoA Reductase Inhibitors. The Role of 21-Methyl in Recognition of HMG-CoA Reductase and the Ileal Bile Acid Transport System

Simultaneous evaluation of HMG‐CoA reductase and cholesterol 7α‐hydroxylase activities by electrospray tandem MS

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