Supplemental Table 1: Mass spectrometer settings and transitions for drug analysis

Compound Electrospray mode Transition Cone (V) Collision energy

Paraxanthine +ve 181.21> 124.11 30 20

Caffeine +ve 195.26 > 138.17 26 23

Bupropion +ve 240.43 > 184.26 28 18

OH-Bupropion +ve 256.40 > 238.31 26 18

Bufuralol +ve 262.37 > 188.33 22 16

1'-OH Bufuralol +ve 278.29 > 186.22 23 22

Midazolam +ve 326.25 > 291.35 46 26

4-OH Midazolam +ve 342.15 > 297.26 37 37

1'-OH Midazolam +ve 342.22 > 324.33 35 25

Triazolam +ve 343.13 > 308.10 55 37

Tolbutamide -ve 269.25 > 169.87 35 17

OH Tolbutamide -ve 285.21 > 185.95 36 17

Chlorzoxazone -ve 168.00 > 131.73 45 21

Deletion of thirty murine cytochrome P450 genes results in viable mice with compromised

drug metabolism

Nico   Scheer,   Lesley   A.   McLaughlin,   Anja   Rode,   A.   Kenneth   MacLeod,   Colin   J.   Henderson   &   C.  

Roland  Wolf    

Supplemental Table 2 Some endogenous substrates P450s in the CYP2C, 2D and 3A gene families P450 Endogenous compounds CYP2C Adrenaline

Arachidonic acid Endothelium-derived hyperpolarizing factor Retinoids Steroid hormones Melatonin

CYP2D Adrenaline Catecholamines Endogenous neurosteroids Melatonin Morphine Tryptamine

CYP3A Arachidonic acid Bile acids Glucocorticoids Oestradiol Prostaglandins Vitamin D Steroid hormones 4β-hydroxycholesterol

Deletion of thirty murine cytochrome P450 genes results in viable mice with

compromised drug metabolism

Nico  Scheer,  Lesley  A.  McLaughlin,  Anja  Rode,  A.  Kenneth  MacLeod,  Colin  J.  Henderson  &  

C.  Roland  Wolf    

Deletion of thirty murine cytochrome P450 genes results in viable mice with compromised drug

metabolism

Nico Scheer, Lesley A. McLaughlin, Anja Rode, A. Kenneth MacLeod, Colin J. Henderson & C.

Roland Wolf

Supplemental Table 3: Pharmacokinetic parameters for metabolites from drug

cocktail administered to WT and Cyp2c/2d/3a KO mice

Metabolite PK parameter WT Cyp2c/2d/3a KO Cyp3a KO

Paraxanthine Cmax (µg/ml) 0.96 ± 0.11 1.1 ± 0.3

Half-life (min) 145.6 ± 40.0 86.1 ± 21.8*

AUClast (min* µg/ml) 233 ± 10 206 ± 42

OH-Bupropion

Cmax (µg/ml) 0.39 ± 0.11 0.65 ± 0.16*

Half-life (min) 285.4 ± 74.4 463.5 ± 539

AUClast (min* µg/ml) 112.8 ± 17.8 188.4 ± 26.5***

OH-Tolbutamide

Cmax (µg/ml) 0.72 ± 0.31 0.18 ± 0.02*

Half-life (min) 421 ± 298 245.5 ± 186.3

AUClast (min* µg/ml) 177.5 ± 31.3 36.5 ± 12.1***

1' OH-Bufuralol

Cmax (µg/ml) 0.21 ± 0.11 0.008 ± 0.003**

Half-life (min) 75 ± 17 144.1 ± 31.8

AUClast (min* µg/ml) 18.5 ± 5.7 1.5 ± 0.5***

1' OH-Midazolam

Cmax (µg/ml) 0.46 ± 0.09 0.1 ± 0.01*** 0.32 ± 0.08##

Half-life (min) 204.4 ± 93.3 234.6 ± 102.2 127.9 ± 23.3

AUClast (min* µg/ml) 73 ± 15 24.2 ± 5.1*** 47.7 ± 21.9

A five-drug cocktail was administered to WT Cyp3a KO and Cyp2c/2d/3a KO mice, blood samples

taken for analysis and metabolite levels determined as detailed in Materials and Methods.

Pharmacokinetic parameters were derived from the WinNonLin software package, and data is shown as

mean +/- SD.

Statistical significance – WT to Cyp2c/2d/3a KO mice: * p < 0.05; ** p < 0.01; *** p < 0.005.

Cyp3a KO to Cyp2c/2d/3a KO mice: ## p < 0.01;