Design, Parallel Synthesis, and Crystal Structures of Pyrazinone Antithrombotics as Selective Inhibitors of the Tissue Factor VIIa Complex

Parlow, John J.; Case, Brenda L.; Dice, Thomas A; Fenton, Ricky L; Hayes, Michael J.; Jones, Darin E.; Neumann, William L.; Wood, Rhonda S.; Lachance, Rhonda M.; Girard, Thomas; Nicholson, Nancy S.; Clare, Michael; Stegeman, Roderick A.; Stevens, Anna M.; Stallings, William C.; Kurumbail, Ravi G.; South, Michael S.

doi:10.1021/jm030131l

Cited by 77 publications

(44 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The coumarin skeleton was chosen as a fluorescent chromophore, and an azidophenyl moiety was introduced as the photoreactive part. [16][17][18] The probe CAFSA (6) was synthesized as shown in Fig. 9.…”

Section: Resultsmentioning

confidence: 99%

Fusarielin A as an Anti-angiogenic and Anti-proliferative Agent: Basic Biological Characterization

Fujimoto

Aoyama

Noguchi‐Yachide

et al. 2008

CHEMICAL & PHARMACEUTICAL BULLETIN

View full text Add to dashboard Cite

show abstract

Section: Resultsmentioning

confidence: 99%

Fusarielin A as an Anti-angiogenic and Anti-proliferative Agent: Basic Biological Characterization

Fujimoto

Aoyama

Noguchi‐Yachide

et al. 2008

CHEMICAL & PHARMACEUTICAL BULLETIN

View full text Add to dashboard Cite

show abstract

“…Kinetics of rTF⅐FVIIa Inhibition by G17905-Enzyme kinetic studies with a substituted benzamidine indicated that FVIIa active site inhibitors are competitive with respect to small peptidyl substrates and noncompetitive with respect to macromolecular substrate activation (70). So far, all published substituted benzamidine inhibitors of FVIIa bind to the standard active site conformation (71)(72)(73)(74)(75)(76). However, G17905 binds to the nonstandard active site conformation, raising the possibility that G17905 could follow different inhibition kinetics.…”

Section: Resultsmentioning

confidence: 99%

A Selective, Slow Binding Inhibitor of Factor VIIa Binds to a Nonstandard Active Site Conformation and Attenuates Thrombus Formation in Vivo

Olivero¹,

Eigenbrot

Goldsmith³

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

The serine protease factor VIIa (FVIIa) in complex with its cellular cofactor tissue factor (TF) initiates the blood coagulation reactions. TF⅐FVIIa is also implicated in thrombosis-related disorders and constitutes an appealing therapeutic target for treatment of cardiovascular diseases. To this end, we generated the FVIIa active site inhibitor G17905, which displayed great potency toward TF⅐FVIIa (K i ‫؍‬ 0.35 ؎ 0.11 nM). G17905 did not appreciably inhibit 12 of the 14 examined trypsin-like serine proteases, consistent with its TF⅐FVIIa-specific activity in clotting assays. The crystal structure of the FVIIa⅐G17905 complex provides insight into the molecular basis of the high selectivity. It shows that, compared with other serine proteases, FVIIa is uniquely equipped to accommodate conformational disturbances in the Gln 217 -Gly 219 region caused by the ortho-hydroxy group of the inhibitor's aminobenzamidine moiety located in the S1 recognition pocket. Moreover, the structure revealed a novel, nonstandard conformation of FVIIa active site in the region of the oxyanion hole, a "flipped" Lys 192 -Gly 193 peptide bond. Macromolecular substrate activation assays demonstrated that G17905 is a noncompetitive, slow-binding inhibitor. Nevertheless, G17905 effectively inhibited thrombus formation in a baboon arterio-venous shunt model, reducing platelet and fibrin deposition by ϳ70% at 0.4 mg/kg ؉ 0.1 mg/kg/ min infusion. Therefore, the in vitro potency of G17905, characterized by slow binding kinetics, correlated with efficacious antithrombotic activity in vivo.

show abstract

“…Using the crystal structure of TF/VIIa bound to a tripeptide-␣-ketothiazole, hundreds of pyrazinone TF/VIIa inhibitors were prepared and tested in several serine protease enzyme assays (Parlow et al, 2003;South et al, 2003). The structure activity relationship of the pyrazinone inhibitors was used to develop PHA-927F, a potent, selective, nonpeptide inhibitor of TF/ VIIa.…”

mentioning

confidence: 99%

Pharmacological Interruption of Acute Thrombus Formation with Minimal Hemorrhagic Complications by a Small Molecule Tissue Factor/Factor VIIa Inhibitor: Comparison to Factor Xa and Thrombin Inhibition in a Nonhuman Primate Thrombosis Model

Suleymanov¹,

Szalony²,

Salyers³

et al. 2003

J Pharmacol Exp Ther

Self Cite

View full text Add to dashboard Cite

This study was designed to evaluate the antithrombotic efficacy and bleeding propensity of a selective, small-molecule inhibitor of tissue factor/factor VIIa (TF/VIIa) in comparison to smallmolecule, selective inhibitors of factor Xa and thrombin in a nonhuman primate model of thrombosis. Acute, spontaneous thrombus formation was induced by electrolytic injury to the intimal surface of a femoral blood vessel, which results in thrombus propagation at the injured site. The TF/FVIIa inhibitor 3-amino-5-[1- [2-({4-[amino(imino)methyl]benzyl}amino)-2-oxoethyl]-3-chloro-5-(isopropylamino)-6-oxo-1,6-dihydropyrazin-2-yl]benzoic acid dihydrochloride (PHA-927F) was fully effective in prevention of thrombosis-induced vessel occlusion at a dose of 400 g/kg/min, i.v., in the arterial vasculature (femoral artery). Neither the effective dose nor multiples up to 4.4-fold the effective arterial plasma concentration elicited any significant effect on bleeding time or blood loss from either the bleeding time site or the surgical (femoral isolation) site. Smallmolecule inhibitors of factor Xa or thrombin were effective arterial antithrombotic agents; however, in contrast to the TF/ FVIIa inhibitor, they both elicited substantial increases in bleeding propensity at the effective dose and at multiples of the effective plasma concentration. These data indicate that TF/ VIIa inhibition effectively prevented arterial thrombosis with less impact on bleeding parameters than equivalent doses of factor Xa and thrombin inhibitors.The most common cause of mortality in the United States and Western countries is cardiovascular disease, predominantly associated with acute coronary syndromes consisting of unstable angina, myocardial infarction, and sudden death (Braunwald et al., 1989). Acute coronary syndromes are associated with acute thrombus formation, often as the result of rupture of the thin fibrous cap of a vulnerable plaque. Upon plaque rupture, platelet activation occurs followed by a cascade of coagulation reactions as tissue factor is exposed to soluble factor VIIa in the blood (Moreno et al., 1996).Tissue factor initiates the extrinsic pathway of blood coagulation and is the obligate cofactor for activation of zymogen coagulation factor VII to the serine protease factor VIIa. The tissue factor/factor VIIa complex (TF/VIIa) initiates coagulation by activating the physiological substrates factor IX and factor X, ultimately leading to thrombin generation and fibrin deposition. Tissue factor is a cell surface-expressed glycoprotein that is composed of a 219-amino acid residue extracellular domain, a single transmembrane sequence, and a short cytoplasmic domain (Edgington et al., 1991). Tissue factor is primarily expressed in the media and adventitia of blood vessels forming a hemostatic envelope that surrounds the vessel.Following plaque rupture or vascular damage, such as occurs in hypertension, atherosclerosis, diabetes, smoking, and interventional procedures, such as balloon angioplasty, tissue factor is exposed to the blood a...

show abstract

Design, Parallel Synthesis, and Crystal Structures of Pyrazinone Antithrombotics as Selective Inhibitors of the Tissue Factor VIIa Complex

Cited by 77 publications

References 61 publications

Fusarielin A as an Anti-angiogenic and Anti-proliferative Agent: Basic Biological Characterization

Fusarielin A as an Anti-angiogenic and Anti-proliferative Agent: Basic Biological Characterization

A Selective, Slow Binding Inhibitor of Factor VIIa Binds to a Nonstandard Active Site Conformation and Attenuates Thrombus Formation in Vivo

Pharmacological Interruption of Acute Thrombus Formation with Minimal Hemorrhagic Complications by a Small Molecule Tissue Factor/Factor VIIa Inhibitor: Comparison to Factor Xa and Thrombin Inhibition in a Nonhuman Primate Thrombosis Model

Contact Info

Product

Resources

About