. Author manuscript; available in PMC: 2014 Jun 27.

Published in final edited form as: J Med Chem. 2013 Jun 13;56(12):5059–5070. doi: 10.1021/jm400369q

Table 4.

Inhibition of recombinant wild-type and mutant thrombins by trimer 9a.^a

Thrombin	log IC₅₀	HS	IC₅₀ (μM)	ΔY^a (%)
rWT	−6.58 ± 0.02	3.4 ± 0.5^b	0.26 ± 0.01	49 ± 2
R93A	−6.73 ± 0.04	1.8 ± 0.3	0.19 ± 0.02	66 ± 6
R97A	−6.44 ± 0.04	2.6 ± 0.5	0.37 ± 0.03	47 ± 3
R101A	−6.41 ± 0.07	2.1 ± 0.6	0.39 ± 0.06	55 ± 6
R126A	−6.55 ± 0.07	1.8 ± 0.5	0.29 ± 0.05	65 ± 11
R165A	−6.59 ± 0.11	1.1 ± 0.3	0.26 ± 0.07	61 ± 8
K169A	−6.48 ± 0.03	2.1 ± 0.2	0.33 ± 0.02	59 ± 3
R173A	−6.59 ± 0.04	2.4 ± 0.7	0.26 ± 0.03	62 ± 8
R175A	−6.363 ± 0.04	5.2 ± 2.1	0.43 ± 0.04	37 ± 3
R233A	−6.01 ± 0.04	3.1 ± 0.6	0.98 ± 0.10	44 ± 2
K235A	−6.57 ± 0.02	2.0 ±0.1	0.27 ± 0.01	50 ± 1
K236A	−6.40 ± 0.01	3.0 ± 0.3	0.40 ± 0.04	52 ± 1
K240A	−6.57 ± 0.04	2.3 ± 0.4	0.27 ± 0.03	49 ± 3

Inhibition studies were performed in 20 mM Tris-HCl buffer, pH 7.4, containing 100 mM NaCl, 2.5 mM CaCl₂ and 0.1 % PEG 8000 in PEG20000-coated acrylic cuvettes. S-2366 was used as thrombin substrate and trimer 9a was incubated overnight with the enzyme at 25 °C. Logistic equation 1 was used to fit the dose-dependence of the residual thrombin activity to obtain log IC₅₀, HS, Y₀, and Y_M. ΔY = Y_M − Y₀. See ‘Experimental Procedures’ for details.

Represents 1 ± S.E.