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PDBsum entry 1ygc
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References listed in PDB file
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Key reference
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Title
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A selective, Slow binding inhibitor of factor viia binds to a nonstandard active site conformation and attenuates thrombus formation in vivo.
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Authors
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A.G.Olivero,
C.Eigenbrot,
R.Goldsmith,
K.Robarge,
D.R.Artis,
J.Flygare,
T.Rawson,
D.P.Sutherlin,
S.Kadkhodayan,
M.Beresini,
L.O.Elliott,
G.G.Deguzman,
D.W.Banner,
M.Ultsch,
U.Marzec,
S.R.Hanson,
C.Refino,
S.Bunting,
D.Kirchhofer.
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Ref.
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J Biol Chem, 2005,
280,
9160-9169.
[DOI no: ]
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PubMed id
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Abstract
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The serine protease factor VIIa (FVIIa) in complex with its cellular cofactor
tissue factor (TF) initiates the blood coagulation reactions. TF.FVIIa 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 TF.FVIIa (Ki = 0.35 +/- 0.11 nM). G17905 did not appreciably inhibit 12
of the 14 examined trypsin-like serine proteases, consistent with its
TF.FVIIa-specific activity in clotting assays. The crystal structure of the
FVIIa.G17905 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
Gln217-Gly219 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" Lys192-Gly193 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 approximately 70% 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.
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Figure 1.
FIG. 1. A, synthesis key intermediates. a, (i) LiCl,
N,N-dimethylformamide 150 °C; (ii) BnCl, K[2]CO[3]. b, H[2],
Pd/C, MeOH. c, t-butyldi-methylsilylchloride, imidazole. d (i)
t-BuLi, B(OMe)[3], (ii) H[2]O[2]/AcOH. e, Cs[2]CO[3], EtI,
N,N-dimethylformamide. f, t-BuLi, N,N-dimethylformamide. B,
synthesis of G17905 [GenBank]
. a, tosylmethylisocyanide, BF[3]OEt[2], MeOH/H[2]O. b, LiOH,
tetrahydrofuran/H[2]O. c, CDI, m-nitrobenzene sulfonamide, DBU,
tetrahydrofuran. d, H[2]/C/Pd. e, (i) EtOH, HCl; (ii) MeOH,
NH[3]. f, reverse phase chiral chromatographic separation,
S-WELK-O column, isopropyl alcohol, pH 5.5.
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Figure 2.
FIG. 2. First generation sulfonamide- and
acylsulfonamide-based FVIIa inhibitors.
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The above figures are
reprinted
by permission from the ASBMB:
J Biol Chem
(2005,
280,
9160-9169)
copyright 2005.
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