Dibasic Benzo[<i>b</i>]thiophene Derivatives as a Novel Class of Active Site-Directed Thrombin Inhibitors. 1. Determination of the Serine Protease Selectivity, Structure−Activity Relationships, and Binding Orientation

Sall, Daniel J.; Bastian, Jolie A.; Briggs, Stephen L.; Buben, John A.; Chirgadze, Nickolay Y.; Clawson, David K.; Denney, Michael L.; Giera, Deborah D.; Gifford‐Moore, Donetta S.; Harper, Richard W.; Hauser, Kenneth L.; Klimkowski, V. J.; Kohn, Todd J.; Lin, Hung‐Yu; McCowan, Jefferson R.; Palkowitz, Alan D.; Smith, Gary; Takeuchi, Kumiko; Thrasher, K. Jeff; Tinsley, Jennifer M.; Utterback, Barbara G.; Yan, S. Betty; Zhang, Minsheng

doi:10.1021/jm9704107

Cited by 53 publications

(27 citation statements)

References 11 publications

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“…The apparent K ass values were obtained in a high-volume testing protocol [13][14][15] using automated dilutions of inhibitors (performed in triplicate at 4-8 inhibitor concentrations) into 96-well plates and chromogenic substrate hydrolysis rates determined at 405 nm using a Thermomax plate reader from Molecular Devices (San Francisco, CA). The assay protocol was 50 µl buffer (0.06 M tris, 0.3 M NaCl, pH 7.4), 25 µl inhibitor test solution (in MeOH), 25 µl human fXa (32 nM in 0.03 M tris, 0.15 M NaCl, 1 mg/ml HSA), and, finally, 150 µl substrate (0.3 mM in water) added within 2 min to start hydrolysis.…”

Section: Binding Affinity For Fxamentioning

confidence: 99%

The Minimum Significant Ratio: A Statistical Parameter to Characterize the Reproducibility of Potency Estimates from Concentration-Response Assays and Estimation by Replicate-Experiment Studies

Farmen²,

et al. 2006

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The authors show by illustration that procedures used to validate the reliability of single-concentration high-throughput screens such as the signal window and Z′ factor do not ensure sufficient reliability in potency estimates from concentration response assays. They develop the minimum significant ratio as a statistical parameter to characterize the fold change between 2 compounds run in the same experiment that can be considered a real difference and use this parameter to characterize the reliability of the assay. They adapt methods described by Bland and Altman to develop a simple set of 2 experiments to estimate the minimum significant ratio and show that this protocol can identify assays that lack reproducibility. The methods are then extended to validate the equivalency of the same assay run by multiple laboratories. (Journal of Biomolecular Screening 2006:253-261)

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Section: Binding Affinity For Fxamentioning

confidence: 99%

The Minimum Significant Ratio: A Statistical Parameter to Characterize the Reproducibility of Potency Estimates from Concentration-Response Assays and Estimation by Replicate-Experiment Studies

Farmen²,

et al. 2006

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“…Indeed, several potent inhibitors have been identified that include P1 groups such as aminoalkyl, aminocyclohexyl [137,187,188], aminopyridine [32,138,153,185,189], aminopyridazine [185], aminopyrimidine [185], aminopyrazine [185], benzamidrazone [190,191], thienylamidine [133], azaphenylalanine [192], 4,5,6,7-tetrahydro-1,3-benzothiazol-2-amine, 4,5,6,7-tetrahydro-2H-indazole and 2-imino-4,5,6,7-tetrahydro-1,3-benzothiazol-3(2H)-yl-amine [193]. Furthermore, there are also a few examples of neutral P1 moieties such as benzothiophene and hydroxybenzothiophene [194][195][196], cyclohexylmethyl, benzyl and dichlorobenzyl [197], 3-chlorobenzyl and 2-oxyacetamide-5-chlorobenzyl [38,198], phenol [33], tryptophan [199] and 6-fluorotryptamine [200]. As one of the most recent examples, screening of a fragment-based compound library also resulted in the discovery of thrombin-binding fragments with a nonbasic P1 motif [201].…”

Section: New Inhibitors -Old Principlesmentioning

confidence: 99%

Direct thrombin inhibitors – a survey of recent developments

Schwienhorst¹

2006

Cell. Mol. Life Sci.

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Thrombin is a plasma serine protease that plays a key role in coagulation and hemostasis but also in thromboembolic diseases. Direct thrombin inhibitors could, therefore, be beneficial for future anticoagulant therapy in the prophylaxis of venous and arterial thrombosis as well as myocardial infarction. However, development of direct thrombin inhibitors has brought researchers more heartache than success. The most recent setback came this year when AstraZeneca withdrew Ximelagatran, the first orally bioavailable direct thrombin inhibitor that had received regulatory approval (France, 2003), after reports of serious hepatoxicity in a fraction of patients. This review describes the status of direct thrombin inhibitors, focusing on drug candidates that are at present in clinical trials. In addition, some more recent research strategies in the design of novel direct thrombin inhibitors are discussed, which may very well contribute to future developments of potent anticoagulants.

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“…Thiophene 41 was identified at Lilly as a weak active site inhibitor after broad-based screening. 82 The initial empirical structure optimization led to 42 which was 35-fold more potent. Compound 42 has good selectivity versus bovine trypsin and other serine proteases and was modestly bioavailable in rats.…”

Section: New Templatesmentioning

confidence: 99%

Small, noncovalent serine protease inhibitors

Sanderson

1999

Med. Res. Rev.

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Dibasic Benzo[b]thiophene Derivatives as a Novel Class of Active Site-Directed Thrombin Inhibitors. 1. Determination of the Serine Protease Selectivity, Structure−Activity Relationships, and Binding Orientation

Cited by 53 publications

References 11 publications

The Minimum Significant Ratio: A Statistical Parameter to Characterize the Reproducibility of Potency Estimates from Concentration-Response Assays and Estimation by Replicate-Experiment Studies

The Minimum Significant Ratio: A Statistical Parameter to Characterize the Reproducibility of Potency Estimates from Concentration-Response Assays and Estimation by Replicate-Experiment Studies

Direct thrombin inhibitors – a survey of recent developments

Small, noncovalent serine protease inhibitors

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