Table 4.
Distances of Protons in Flurbiprofen and Dapsone in CYP2C9 Variants.a
| Compound | Proton Resonanceb | CYP2C9.1c | CYP2C9.2d | CYP2C9.3e | CYP2C9.5f | ||||
|---|---|---|---|---|---|---|---|---|---|
| r (||, Å)g | r(|, Å)h | r (||, Å) | r(|, Å) | r (||, Å) | r(|, Å) | r (||, Å) | r(|, Å) | ||
| CH3 | 6.77 (0.06) | 7.61 (0.06) | 5.71 (0.10) | 6.42 (0.11) | 6.54 (0.11) | 7.34 (0.12) | 5.94 (0.10) | 6.66 (0.11) | |
| 2 | 6.35 (0.01) | 7.13 (0.02) | 6.09 (0.37) | 6.84 (0.42) | 6.46 (0.20) | 7.25 (0.22) | 6.06 (0.20) | 6.80 (0.22) | |
| 5 | 6.77 (0.15) | 7.60 (0,18 | 5.66 (0.46) | 6.35 (0.52) | 6.27 (0.17) | 7.05 (0.19) | 5.57 (0.19) | 6.26 (0.21) | |
| Flurbiprofen | 6 | 6.33 (0.10) | 7.10 (0.11) | 6.05 (0.34) | 6.80 (0.38) | 6.22 (0.14) | 6.98 (0.16) | 6.44 (0.20) | 7.23 (0.22) |
| 2′,6′ | 6.43 (0.08) | 7.21 (0.10) | 6.07 (0.22) | 6.81 (0.25) | 6.30 (0.11) | 7.07 (0.13) | 5.84 (0.10) | 6.55 (0.11) | |
| 3′,5′ | 6.48 (0.09) | 7.27 (0.10) | 5.54 (0.22) | 6.22 (0.25) | 6.25 (0.12) | 7.02 (0.14) | 5.76 (0.10) | 6.47 (0.11) | |
| 4′ | 6.62 (0.27) | 7.44 (0.30 | 5.76 (0.45) | 6.47 (0.51) | 6.29 (0.24) | 7.06 (0.27) | 5.90 (0.20) | 6.62 (0.22) | |
| 2,2′,6,6′ | 5.84 (0.07)i | 6.55 (0.08)k | 6.28 (0.09) | 7.05 (0.10) | 7.10 (0.09) | 7.98 (0.10) | 6.11 (0.08) | 6.86 (0.10) | |
| Dapsone | 6.55 (0.08)j | 7.36 (0.09)l | 7.05 (0.10) | 7.91 (0.11) | 7.98 (0.10) | 8.96 (0.12) | 6.86 (0.10) | 7.70 (0.12) | |
| 3,3′,5,5′ | 6.06 (0.01)i | 6.81 (0.11)k | 6.31 (0.10) | 7.08 (0.11) | 7.07 (0.11) | 7.94 (0.13) | 6.24 (0.11) | 7.01 (0.13) | |
| 6.81 (0.11)j | 7.64 (0.12)l | 7.08 (0.11) | 7.95 (0.13) | 7.94 (0.13) | 8.91 (0.14) | 7.01 (0.13) | 7.87 (0.14) | ||
| CH3 | 6.21 (0.06) | 6.97 (0.06) | 5.71 (0.12) | 6.40 (0.13) | 6.10 (0.13) | 6.85 (0.14) | 5.25 (0.08) | 5.90 (0.09) | |
| 2 | 5.69 (0.06) | 6.39 (0.12) | 5.86 (0.49) | 6.58 (0.55) | 5.74 (0.16) | 6.45 (0.18) | 5.28 (0.16) | 5.92 (0.18) | |
| 5 | 5.84 (0.29) | 6.56 (0.33) | 5.82 (0.58) | 6.54 (0.65) | 6.40 (0.21) | 7.19 (0.24) | 5.16 (0.14) | 5.79 (0.15) | |
| 6 | 6.00 (0.21) | 6.74 (0.24) | 6.79 (0.48) | 7.62 (0.53) | 6.10 (0.21) | 6.84 (0.23) | 5.43 (0.14) | 6.10 (0.15) | |
| 2′,6′ | 5.90 (0.18) | 6.62 (0.20) | 5.68 (0.20) | 6.37 (0.23) | 6.04 (0.11) | 6.78 (0.12) | 5.37 (0.07) | 6.03 (0.08) | |
| Flurbiprofen and Dapsone | 3′,5′ | 5.85 (0.18) | 6.56 (0.20) | 5.70 (0.37) | 6,49 (0.42) | 5.81 (0.13) | 6.52 (0.15) | 5.33 (0.09) | 5.98 (0.11) |
| 4′ | 5.61 (0.23) | 6.29 (0.26) | 5.68 (0.48) | 6.37 (0.58) | 5.66 (0.27) | 6.35 (0.30) | 5.20 (0.23) | 5.83 (0.26) | |
| 2,2′,6,6′ | 5.82 (0.04)i | 6.53 (0.05)k | 6.17 (0.08) | 6.92 (0.10) | 6.02 (0.08) | 6.76 (0.09) | 5.46 (0.08) | 6.13 (0.09) | |
| 6.53 (0.05)j | 7.33 (0.06)l | 6.92 (0.10) | 7.77 (0.11) | 6.76 (0.09) | 7.59 (0.11) | 6.13 (0.09) | 6.88 (0.11) | ||
| 3,3′,5,5′ | 6.11 (0.06)i | 6.86 (0.07)k | 6.07 (0.08) | 6.81 (0.13) | 6.06 (0.12) | 6.80 (0.13) | 5.57 (0.12) | 6.26 (0.13) | |
| 6.86 (0.07)j | 7.70 (0.08)l | 6.81 (0.13) | 7.65 (0.15) | 6.80 (0.13) | 7.64 (0.15) | 6.26 (0.13) | 7.02 (0.15) | ||
Errors for measurements were < 10%.
See Figure 1 for numbering scheme of the protons for flurbiprofen and dapsone. T1 times for the HC-CO2H proton could not be accurately determined due to interference from the residual glycerol resonances.
[P450 2C9.1] = 0.016 μM, [Flurbiprofen] = 160 μM; [Dapsone] = 100 μM. αM = [P450]/(KS+[substrate]), KS (flurbiprofen) = 16.0 μM, αM (Flurbiprofen) = 9.15×10−5, KS (dapsone) = 100 μM (not obtainable by UV spectroscopy, assumed KS=KM), αM (Dapsone) = 7.00×10−5, αM (Flurbiprofen and Dapsone) = 3.72×10−5, S(S+1) = 2.07 (Flurbiprofen), 1.65 (Dapsone), 2.07 (Flurbiprofen+Dapsone).
[P450 2C9.2] = 0.005 μM, [Flurbiprofen] = 50 μM; [Dapsone] = 100 μM. αM = [P450]/(KS+[substrate]), KS (flurbiprofen) = 4.5 μM, αM (Flurbiprofen) = 9.09×10−5, KS (dapsone) = 100 μM (not obtainable by UV spectroscopy, assumed KS=KM), αM (Dapsone) = 7.00×10−5, αM (Flurbiprofen and Dapsone) = 1.96×10−5, S(S+1) = 1.64 (Flurbiprofen), 1.17 (Dapsone), 2.81 (Flurbiprofen+Dapsone).
[P450 2C9.2] = 0.014 μM, [Flurbiprofen] = 140 μM; [Dapsone] = 100 μM. αM = [P450]/(KS+[substrate]), KS (flurbiprofen) = 14 μM, αM (Flurbiprofen) = 9.09×10−5, KS (dapsone) = 100 μM (not obtainable by UV spectroscopy, assumed KS=KM), αM (Dapsone) = 7.00×10−5, αM (Flurbiprofen and Dapsone) = 3.95×10−5, S(S+1) = 1.78 (Flurbiprofen), 1.00 (Dapsone), 2.71 (Flurbiprofen+Dapsone).
[P450 2C9.2] = 0.016 μM, [Flurbiprofen] = 160 μM; [Dapsone] = 100 μM. αM = [P450]/(KS+[substrate]), KS (flurbiprofen) = 16 μM, αM (Flurbiprofen) = 9.09×10−5, KS (dapsone) = 100 μM (not obtainable by UV spectroscopy, assumed KS=KM), αM (Dapsone) = 7.00×10−5, αM (Flurbiprofen and Dapsone) = 3.72×10−5, S(S+1) = 1.56 (Flurbiprofen), 1.32 (Dapsone), 1.83 (Flurbiprofen+Dapsone).
r(||) = [9.87×1016 * T1P * αM * S(S+1) * τC)]1/6, τC = 2 × 10−10 [18] and corresponds to the parallel binding orientation of flurbiprofen.
r(|′) = [9.87×1016 * 2T1P * αM * S(S+1) * τC)]1/6, τC = 2 × 10−10 [18] and corresponds to the perpendicular binding orientation of flurbiprofen.
r(||) = [9.87×1016 * T1P * αM * S(S+1) * τC)]1/6, τC = 2 × 10−10 [18] and corresponds to the symmetrical/parallel binding orientation of dapsone.
r(||) = [9.87×1016 * 2T1P * αM * S(S+1) * τC)]1/6, τC = 2 × 10−10 [18].and corresponds to the unsymmetrical/parallel binding orientation of dapsone.
r(|) = [9.87×1016 * 2T1P * αM * S(S+1) * τC)]1/6, τC = 2 × 10−10 [18] and corresponds to the symmetrical/perpendicular binding orientation of dapsone.
r(|) = [9.87×1016 * 4T1P * αM * S(S+1) * τC)]1/6, τC = 2 × 10−10 [18] and corresponds to the unsymmetrical/perpendicular binding orientation of dapsone.