Cytotoxicity of ionic liquids and precursor compounds towards human cell line HeLa

Cytotoxcty of oc lqud ad precuror compoud toward huma cell le HeLa Xuefeg Wag, a,b C. Adré Ohl, a Qghua Lu,* a Zhaofu Fe, c Ju Hu, b ad Paul J. Dyo c a School of Chemtry ad Chemcal Techology, Shagha Jao Tog Uverty, Shagha 200240, P.R. Cha Fax:86-02-54747535; Tel: 86-02-54747535; E-mal: qhlu@jtu.edu.c b School of Lfe Scece ad Botechology, Shagha Jao Tog Uverty, Shagha 200240, P.R. Cha. Tel: 86-02-34203723; E-mal: xuefegwag@jtu.edu.c c Laboratory of Orgaometallc ad Medccal Chemtry, Ittut de Scece et Igéere Chmque, Sw Federal Ittute of Techology, 05-Lauae, Swtzerlad. Tel.: 4- (02-6939838; E-mal: zhaofu.fe@epfl.ch. Idex a. Toxcty of ome commo olvet ad alt toward HeLa cell. b. Detaled parameter for alt ued cocetrato addto expermet. 2. Reult from cocetrato addto expermet. 3. K-value calculato for lthum bromde ad chole bromde. 4. Flow cytometrc meauremet of mtochodral membrae potetal ad reactve oxyge pece HeLa cell expoed to [C 2 mm][bf 4 ]. 5. Dervato of equato 3a ad 3b the ma text.

a. Toxcty of ome commo olvet ad alt toward HeLa cell. Table. Toxcty of ome commo olvet ad alt toward HeLa cell, determed after 48 hour the preece of FBS. Compoud f(x0-4 b EC 50 ± SE a (mm R 2 Acetoe - b.4 449 ± 69 0.97 Acetotrle - - >2900 - Ethaol - 2.92 866 ± 37 0.99 N,N-Dmethylformamde - 3.92 03 ± 8 0.99 Dmethylulfoxde - 3.50 449 ± 56 0.98 Sodum dodecylbezeeulfoate - 4.79 0.8 ± 0.0 0.99 Sodum chlorde 54.2 2.0 0 ± 32 0.95 Sodum bromde 0.3 5.04 70 ± 6 0.97 Lthum chlorde 0 2.4 4.02 ± 2.49 0.99 Lthum bromde -.59 28.82 ± 5.9 0.95 a Stadard error. b f wa fxed a zero. b. Detaled parameter for alt ued cocetrato addto expermet. Table 2. Detaled parameter for alt ued cocetrato addto expermet. Toxcte determed the abece of FBS after 24 hour. Cato -R Ao f ± SE f (x0-4 b ± SE b EC 50 ± SE EC50 (mm a R 2 -C 2 H 5 Br - - 2.89 ± 0.22 9.60 ± 0.35 4 0.99 -C 4 H 9 Br - - 0.93 ± 0.09 2.6 ± 0.23 6 0.98 -C 8 H 7 Br - -.09 ± 0.06 0.02 ± 0.00 5 0.99 -CH 2 C 6 H 5 Br - - 0.87 ± 0.05 0.55 ± 0.03 6 0.99 2 -C 2 H 5 Br - -.24 ± 0.6 3.85 ±.34 6 0.98 2 -C 4 H 9 Br - - 0.83 ± 0.2.74 ± 0.28 6 0.97 2 -C 8 H 7 Br - -.29 ± 0.0 0.04 ± 0.003 4 0.99 2 -CH 2 C 6 H 5 Br - -.03 ± 0.0 0.53 ± 0.05 4 0.99 3 -CH 3 Br - 0.74 ± 0.4 4.09 ± 0.53 69.57 ± 2.94 6 0.99 3 -C 2 H 5 Br - 0.23 ± 0.4 4.69 ± 0.6 53.23 ±.70 6 0.99 3 -C 4 H 9 Br - 0.7 ± 0.60 2.52 ± 0.44 25.23 ± 2.23 4 0.98 3 -C 8 H 7 Br - -.42 ± 0.08 0.20 ± 0.0 6 0.99 3 -CH 2 C 6 H 5 Br - -.2 ± 0.4 2.87 ± 0.3 4 0.98 L - Br - 0.97 ± 0.29 3.24 ± 0.2 32.73 ± 0.9 6 0.99 a Sample ze. See ecto 5. 2

2. Reult from cocetrato addto expermet Table 3. Cocetrato addto data for mxture of alt ad cell cultured the abece of FBS for 24 hour. Etry Mxture C a b /EC 50, [C 2 Chol]Br ( ad [C 4 Chol]Br (2 2 [C 2 Chol]Br ( ad [C 8 Chol]Br (2 3 [C 2 Chol]Br ( ad [C 6 H 5 CH 2 Chol]Br (2 4 [C 4 Chol]Br ( ad[ C 8 Chol]Br (2 5 [C 4 Chol]Br ( ad [C 6 H 5 CH 2 Chol]Br (2 C 2 /EC 50,2 P c ± SE P -,d P,e.0-0.50 ± 0.04 0.47 0.53 0.9 0. 0.50 ± 0.03 0.47 0.53 0.8 0.2 0.5 ± 0.05 0.46 0.54 0.7 0.3 0.47 ± 0.02 0.45 0.54 0.6 0.4 0.46 ± 0.04 0.45 0.55 0.5 0.5 0.45 ± 0.03 0.44 0.55 0.4 0.6 0.49 ± 0.02 0.43 0.56 0.3 0.7 0.50 ± 0.0 0.43 0.57 0.2 0.8 0.46 ± 0.0 0.42 0.57 0. 0.9 0.50 ± 0.02 0.4 0.58 -.0 0.50 ± 0.02 0.4 0.58.0-0.54 ± 0.03 0.48 0.52 0.9 0. 0.58 ± 0.06 0.48 0.52 0.8 0.2 0.56 ± 0.06 0.48 0.53 0.7 0.3 0.60 ± 0.04 0.47 0.53 0.6 0.4 0.56 ± 0.04 0.47 0.53 0.5 0.5 0.54 ± 0.05 0.47 0.53 0.4 0.6 0.48 ± 0.02 0.47 0.54 0.3 0.7 0.43 ± 0.03 0.47 0.54 0.2 0.8 0.42 ± 0.03 0.46 0.54 0. 0.9 0.45 ± 0.02 0.46 0.54 -.0 0.48 ± 0.02 0.46 0.54.0-0.5 ± 0.03 0.48 0.52 0.9 0. 0.57 ± 0.02 0.47 0.53 0.8 0.2 0.58 ± 0.04 0.46 0.53 0.7 0.3 0.58 ± 0.04 0.46 0.54 0.6 0.4 0.60 ± 0.04 0.45 0.54 0.5 0.5 0.55 ± 0.03 0.44 0.55 0.4 0.6 0.56 ± 0.03 0.44 0.56 0.3 0.7 0.54 ± 0.02 0.43 0.56 0.2 0.8 0.54 ± 0.03 0.42 0.57 0. 0.9 0.50 ± 0.03 0.42 0.57 -.0 0.49 ± 0.03 0.4 0.58.0-0.46 ± 0.03 0.45 0.55 0.9 0. 0.53 ± 0.03 0.45 0.54 0.8 0.2 0.60 ± 0.04 0.45 0.54 0.7 0.3 0.68 ± 0.04 0.46 0.54 0.6 0.4 0.67 ± 0.03 0.46 0.54 0.5 0.5 0.64 ± 0.07 0.46 0.54 0.4 0.6 0.57 ± 0.03 0.46 0.54 0.3 0.7 0.58 ± 0.04 0.47 0.53 0.2 0.8 0.57 ± 0.05 0.47 0.53 0. 0.9 0.56 ± 0.03 0.47 0.53 -.0 0.5 ± 0.0 0.47 0.53.0-0.5 ± 0.0 0.45 0.54 0.9 0. 0.56 ± 0.03 0.45 0.55 0.8 0.2 0.55 ± 0.05 0.45 0.55 3

Etry Mxture C a b /EC 50, C 2 /EC 50,2 P c ± SE P -,d P,e 0.7 0.3 0.57 ± 0.04 0.45 0.55 0.6 0.4 0.56 ± 0.02 0.45 0.55 0.5 0.5 0.58 ± 0.04 0.45 0.55 0.4 0.6 0.56 ± 0.02 0.45 0.55 0.3 0.7 0.58 ± 0.03 0.44 0.55 0.2 0.8 0.54 ± 0.0 0.44 0.55 0. 0.9 0.57 ± 0.02 0.44 0.55 -.0 0.52 ± 0.03 0.44 0.55 6 [C 8 Chol]Br ( ad.0-0.52 ± 0.05 0.48 0.52 [C 6 H 5 CH 2 Chol]Br (2 0.9 0. 0.5 ± 0.0 0.48 0.52 0.8 0.2 0.46 ± 0.02 0.48 0.52 0.7 0.3 0.47 ± 0.02 0.48 0.52 0.6 0.4 0.48 ± 0.02 0.47 0.53 0.5 0.5 0.47 ± 0.0 0.47 0.53 0.4 0.6 0.48 ± 0.03 0.47 0.53 0.3 0.7 0.47 ± 0.03 0.47 0.53 0.2 0.8 0.47 ± 0.0 0.47 0.53 0. 0.9 0.47 ± 0.02 0.46 0.53 -.0 0.49 ± 0.0 0.46 0.54 7 [C 2 Py]Br ( ad.0-0.50 ± 0.03 0.48 0.52 [C 4 Py]Br (2 0.9 0. 0.5 ± 0.02 0.48 0.52 0.8 0.2 0.5 ± 0.05 0.47 0.52 0.7 0.3 0.54 ± 0.02 0.47 0.53 0.6 0.4 0.56 ± 0.03 0.47 0.53 0.5 0.5 0.53 ± 0.03 0.47 0.53 0.4 0.6 0.50 ± 0.05 0.47 0.53 0.3 0.7 0.55 ± 0.04 0.47 0.53 0.2 0.8 0.55 ± 0.08 0.46 0.53 0. 0.9 0.47 ± 0.05 0.46 0.53 8 [C 2 Py]Br ( ad [C 8 Py]Br (2 9 [C 2 Py]Br ( ad [C 6 H 5 CH 2 Py]Br (2 -.0 0.50 ± 0.05 0.46 0.53.0-0.50 ± 0.04 0.47 0.53 0.9 0. 0.6 ± 0.02 0.47 0.53 0.8 0.2 0.66 ± 0.4 0.47 0.53 0.7 0.3 0.64 ± 0.04 0.47 0.53 0.6 0.4 0.64 ± 0.07 0.47 0.52 0.5 0.5 0.63 ± 0.03 0.47 0.52 0.4 0.6 0.5 ± 0.02 0.47 0.52 0.3 0.7 0.56 ± 0.03 0.47 0.52 0.2 0.8 0.58 ± 0.08 0.47 0.52 0. 0.9 0.65 ± 0.0 0.47 0.52 -.0 0.50 ± 0.04 0.47 0.52.0-0.50 ± 0.05 0.47 0.52 0.9 0. 0.54 ± 0.06 0.47 0.52 0.8 0.2 0.56 ± 0.04 0.47 0.52 0.7 0.3 0.57 ± 0.0 0.47 0.52 0.6 0.4 0.56 ± 0.02 0.47 0.53 0.5 0.5 0.59 ± 0.04 0.47 0.53 0.4 0.6 0.63 ± 0.02 0.47 0.53 0.3 0.7 0.62 ± 0.04 0.47 0.53 0.2 0.8 0.60 ± 0.02 0.47 0.53 0. 0.9 0.52 ± 0.04 0.47 0.53 -.0 0.50 ± 0.0 0.47 0.53 0 [C 4 Py]Br ( ad.0-0.50 ± 0.05 0.46 0.53 4

Etry Mxture C /EC 50, C 2 /EC 50,2 P ± SE P -,d P,e [C 8 Py]Br (2 0.9 0. 0.56 ± 0.06 0.46 0.54 0.8 0.2 0.60 ± 0.05 0.46 0.53 0.7 0.3 0.53 ± 0.06 0.46 0.53 0.6 0.4 0.55 ± 0.03 0.46 0.53 0.5 0.5 0.59 ± 0.07 0.47 0.53 0.4 0.6 0.55 ± 0.04 0.47 0.53 0.3 0.7 0.60 ± 0.05 0.47 0.53 0.2 0.8 0.6 ± 0.05 0.47 0.52 0. 0.9 0.50 ± 0.04 0.48 0.52 -.0 0.50 ± 0.05 0.48 0.52 [C 4 Py]Br ( ad.0-0.50 ± 0.03 0.47 0.53 [C 6 H 5 CH 2 Py]Br (2 0.9 0. 0.58 ± 0.05 0.47 0.53 0.8 0.2 0.57 ± 0.07 0.47 0.53 0.7 0.3 0.52 ± 0.06 0.47 0.53 0.6 0.4 0.5 ± 0.05 0.47 0.53 0.5 0.5 0.48 ± 0.06 0.47 0.53 0.4 0.6 0.48 ± 0.06 0.47 0.53 0.3 0.7 0.55 ± 0.05 0.47 0.53 0.2 0.8 0.54 ± 0.06 0.47 0.52 0. 0.9 0.5 ± 0.04 0.47 0.52 2 [C 8 Py]Br ( ad [C 6 H 5 CH 2 Py]Br (2 3 [C 2 mm]br ( ad [C 4 mmbr] (2 4 [C 2 mm]br ( ad [C 8 mm]br (2 -.0 0.50 ± 0.03 0.47 0.52.0-0.50 ± 0.02 0.48 0.52 0.9 0. 0.50 ± 0.0 0.48 0.52 0.8 0.2 0.50 ± 0.04 0.48 0.52 0.7 0.3 0.49 ± 0.03 0.47 0.52 0.6 0.4 0.52 ± 0.02 0.47 0.52 0.5 0.5 0.49 ± 0.0 0.47 0.52 0.4 0.6 0.47 ± 0.0 0.47 0.53 0.3 0.7 0.47 ± 0.03 0.47 0.53 0.2 0.8 0.49 ± 0.05 0.47 0.53 0. 0.9 0.5 ± 0.06 0.47 0.53 -.0 0.50 ± 0.02 0.47 0.53.0-0.50 ± 0.04 0.48 0.52 0.9 0. 0.53 ± 0.05 0.48 0.52 0.8 0.2 0.56 ± 0.06 0.47 0.52 0.7 0.3 0.5 ± 0.04 0.47 0.52 0.6 0.4 0.55 ± 0.03 0.47 0.53 0.5 0.5 0.56 ± 0.04 0.47 0.53 0.4 0.6 0.53 ± 0.06 0.47 0.53 0.3 0.7 0.48 ± 0.02 0.46 0.53 0.2 0.8 0.47 ± 0.03 0.46 0.54 0. 0.9 0.45 ± 0.0 0.46 0.54 -.0 0.5 ± 0.0 0.46 0.54.0-0.54 ± 0.05 0.48 0.52 0.9 0. 0.6 ± 0.04 0.48 0.52 0.8 0.2 0.63 ± 0.02 0.48 0.52 0.7 0.3 0.64 ± 0.02 0.48 0.52 0.6 0.4 0.60 ± 0.03 0.48 0.52 0.5 0.5 0.64 ± 0.03 0.48 0.52 0.4 0.6 0.64 ± 0.05 0.48 0.52 0.3 0.7 0.68 ± 0.03 0.47 0.52 0.2 0.8 0.68 ± 0.04 0.47 0.53 0. 0.9 0.68 ± 0.05 0.47 0.53 5

Etry Mxture C /EC 50, C 2 /EC 50,2 P ± SE P -,d P,e 5 [C 2 mm]br ( ad [C 6 H 5 CH 2 mm]br (2 6 [C 4 mm]br ( ad [C 8 mm]br (2 7 [C 4 mm]br ( ad [C 6 H 5 CH 2 mm]br (2 8 [C 8 mm]br ( ad [C 6 H 5 CH 2 mm]br (2 9 [C 2 Chol]Br ( ad [C 2 Py]Br (2 -.0 0.55 ± 0.02 0.47 0.53.0-0.5 ± 0.0 0.48 0.52 0.9 0. 0.46 ± 0.03 0.48 0.52 0.8 0.2 0.53 ± 0.05 0.48 0.52 0.7 0.3 0.5 ± 0.0 0.48 0.52 0.6 0.4 0.53 ± 0.02 0.48 0.52 0.5 0.5 0.64 ± 0.08 0.48 0.52 0.4 0.6 0.66 ± 0.06 0.48 0.52 0.3 0.7 0.68 ± 0.0 0.48 0.52 0.2 0.8 0.55 ± 0.05 0.48 0.52 0. 0.9 0.54 ± 0.09 0.48 0.52 -.0 0.50 ± 0.0 0.48 0.52.0-0.48 ± 0.05 0.48 0.52 0.9 0. 0.54 ± 0.02 0.48 0.52 0.8 0.2 0.5 ± 0.03 0.48 0.52 0.7 0.3 0.5 ± 0.0 0.48 0.52 0.6 0.4 0.49 ± 0.07 0.48 0.52 0.5 0.5 0.54 ± 0.03 0.48 0.52 0.4 0.6 0.6 ± 0.0 0.48 0.52 0.3 0.7 0.6 ± 0.04 0.48 0.52 0.2 0.8 0.58 ± 0.03 0.48 0.52 0. 0.9 0.56 ± 0.02 0.48 0.5 -.0 0.50 ± 0.0 0.49 0.5.0-0.5 ± 0.0 0.48 0.52 0.9 0. 0.53 ± 0.04 0.48 0.52 0.8 0.2 0.53 ± 0.0 0.48 0.52 0.7 0.3 0.53 ± 0.0 0.48 0.52 0.6 0.4 0.55 ± 0.02 0.48 0.5 0.5 0.5 0.57 ± 0.02 0.49 0.5 0.4 0.6 0.52 ± 0.02 0.49 0.5 0.3 0.7 0.53 ± 0.0 0.49 0.5 0.2 0.8 0.53 ± 0.02 0.49 0.5 0. 0.9 0.57 ± 0.02 0.49 0.5 -.0 0.50 ± 0.02 0.49 0.5.0-0.52 ± 0.0 0.49 0.5 0.9 0. 0.55 ± 0.03 0.49 0.5 0.8 0.2 0.55 ± 0.05 0.49 0.5 0.7 0.3 0.50 ± 0.04 0.49 0.5 0.6 0.4 0.50 ± 0.04 0.49 0.5 0.5 0.5 0.47 ± 0.04 0.49 0.5 0.4 0.6 0.54 ± 0.03 0.49 0.5 0.3 0.7 0.55 ± 0.02 0.49 0.5 0.2 0.8 0.47 ± 0.0 0.49 0.5 0. 0.9 0.47 ± 0.04 0.49 0.5 -.0 0.49 ± 0.02 0.49 0.5.0-0.5 ± 0.0 0.48 0.52 0.9 0. 0.49 ± 0.0 0.47 0.52 0.8 0.2 0.50 ± 0.02 0.47 0.53 0.7 0.3 0.48 ± 0.0 0.47 0.53 0.6 0.4 0.5 ± 0.02 0.46 0.54 0.5 0.5 0.5 ± 0.04 0.46 0.54 0.4 0.6 0.50 ± 0.0 0.45 0.54 0.3 0.7 0.49 ± 0.02 0.45 0.55 6

Etry Mxture C /EC 50, C 2 /EC 50,2 P ± SE P -,d P,e 20 [C 2 Chol]Br ( ad [C 2 mm]br (2 2 [C 2 Py]Br ( ad [C 2 mm]br (2 22 [C 4 Chol]Br ( ad [C 4 Py]Br (2 23 [C 4 Chol]Br ( ad [C 4 mm]br (2 24 [C 4 Py]Br ( ad [C 4 mm]br (2 0.2 0.8 0.50 ± 0.03 0.44 0.55 0. 0.9 0.52 ± 0.02 0.44 0.56 -.0 0.49 ± 0.02 0.43 0.56.0-0.50 ± 0.02 0.47 0.53 0.9 0. 0.49 ± 0.0 0.47 0.53 0.8 0.2 0.52 ± 0.04 0.47 0.53 0.7 0.3 0.50 ± 0.03 0.47 0.53 0.6 0.4 0.49 ± 0.0 0.47 0.53 0.5 0.5 0.53 ± 0.0 0.47 0.53 0.4 0.6 0.54 ± 0.02 0.47 0.53 0.3 0.7 0.54 ± 0.03 0.47 0.53 0.2 0.8 0.56 ± 0.04 0.46 0.53 0. 0.9 0.57 ± 0.02 0.46 0.54 -.0 0.50 ± 0.03 0.46 0.54.0-0.50 ± 0.05 0.46 0.54 0.9 0. 0.5 ± 0.02 0.46 0.54 0.8 0.2 0.50 ± 0.0 0.46 0.54 0.7 0.3 0.49 ± 0.04 0.46 0.53 0.6 0.4 0.48 ± 0.02 0.46 0.53 0.5 0.5 0.50 ± 0.0 0.47 0.53 0.4 0.6 0.53 ± 0.03 0.47 0.53 0.3 0.7 0.52 ± 0.03 0.47 0.53 0.2 0.8 0.53 ± 0.04 0.47 0.52 0. 0.9 0.55 ± 0.03 0.48 0.52 -.0 0.50 ± 0.05 0.48 0.52.0-0.52 ± 0.06 0.46 0.54 0.9 0. 0.5 ± 0.07 0.45 0.54 0.8 0.2 0.5 ± 0.05 0.45 0.54 0.7 0.3 0.47 ± 0.06 0.45 0.55 0.6 0.4 0.44 ± 0.03 0.45 0.55 0.5 0.5 0.48 ± 0.04 0.45 0.55 0.4 0.6 0.52 ± 0.06 0.44 0.55 0.3 0.7 0.48 ± 0.05 0.44 0.55 0.2 0.8 0.49 ± 0.02 0.44 0.55 0. 0.9 0.49 ± 0.03 0.44 0.56 -.0 0.53 ± 0.02 0.43 0.56.0-0.49 ± 0.06 0.46 0.54 0.9 0. 0.45 ± 0.03 0.46 0.54 0.8 0.2 0.47 ± 0.03 0.46 0.54 0.7 0.3 0.47 ± 0.02 0.46 0.54 0.6 0.4 0.47 ± 0.03 0.46 0.54 0.5 0.5 0.50 ± 0.03 0.46 0.54 0.4 0.6 0.50 ± 0.07 0.46 0.54 0.3 0.7 0.5 ± 0.07 0.46 0.54 0.2 0.8 0.5 ± 0.07 0.46 0.54 0. 0.9 0.53 ± 0.04 0.46 0.54 -.0 0.52 ± 0.03 0.46 0.54.0-0.52 ± 0.04 0.46 0.53 0.9 0. 0.5 ± 0.05 0.46 0.53 0.8 0.2 0.48 ± 0.02 0.47 0.53 0.7 0.3 0.50 ± 0.02 0.47 0.53 0.6 0.4 0.50 ± 0.02 0.47 0.52 0.5 0.5 0.50 ± 0.02 0.47 0.52 7

Etry Mxture C /EC 50, C 2 /EC 50,2 P ± SE P -,d P,e 25 [C 8 Chol]Br ( ad [C 8 Py]Br (2 26 [C 8 Chol]Br ( ad [C 8 mm]br (2 27 [C 8 Py]Br ( ad [C 8 mm]br (2 28 [C 6 H 5 CH 2 Chol]Br ( ad [C 6 H 5 CH 2 Py]Br (2 29 [C 6 H 5 CH 2 Chol]Br ( ad [C 6 H 5 CH 2 mm]br (2 0.4 0.6 0.52 ± 0.02 0.47 0.52 0.3 0.7 0.53 ± 0.03 0.48 0.52 0.2 0.8 0.50 ± 0.03 0.48 0.52 0. 0.9 0.54 ± 0.02 0.48 0.52 -.0 0.50 ± 0.03 0.48 0.52.0-0.50 ± 0.0 0.48 0.52 0.9 0. 0.48 ± 0.07 0.48 0.52 0.8 0.2 0.47 ± 0.04 0.48 0.52 0.7 0.3 0.49 ± 0.06 0.48 0.52 0.6 0.4 0.48 ± 0.04 0.48 0.52 0.5 0.5 0.49 ± 0.02 0.48 0.52 0.4 0.6 0.48 ± 0.03 0.48 0.52 0.3 0.7 0.5 ± 0.03 0.48 0.52 0.2 0.8 0.47 ± 0.03 0.48 0.52 0. 0.9 0.49 ± 0.03 0.47 0.52 -.0 0.50 ± 0.03 0.47 0.52.0-0.52 ± 0.04 0.48 0.52 0.9 0. 0.5 ± 0.0 0.48 0.52 0.8 0.2 0.53 ± 0.03 0.48 0.52 0.7 0.3 0.52 ± 0.03 0.48 0.52 0.6 0.4 0.54 ± 0.03 0.48 0.52 0.5 0.5 0.50 ± 0.06 0.48 0.52 0.4 0.6 0.47 ± 0.04 0.48 0.52 0.3 0.7 0.46 ± 0.02 0.48 0.52 0.2 0.8 0.48 ± 0.05 0.48 0.52 0. 0.9 0.47 ± 0.03 0.48 0.52 -.0 0.54 ± 0.03 0.48 0.52.0-0.50 ± 0.03 0.48 0.52 0.9 0. 0.48 ± 0.04 0.48 0.52 0.8 0.2 0.5 ± 0.02 0.48 0.52 0.7 0.3 0.52 ± 0.05 0.48 0.52 0.6 0.4 0.52 ± 0.02 0.48 0.52 0.5 0.5 0.5 ± 0.04 0.48 0.52 0.4 0.6 0.49 ± 0.07 0.48 0.52 0.3 0.7 0.47 ± 0.05 0.48 0.52 0.2 0.8 0.5 ± 0.08 0.48 0.52 0. 0.9 0.50 ± 0.06 0.48 0.52 -.0 0.50 ± 0.02 0.48 0.52.0-0.50 ± 0.04 0.47 0.53 0.9 0. 0.5 ± 0.03 0.47 0.53 0.8 0.2 0.5 ± 0.06 0.47 0.53 0.7 0.3 0.52 ± 0.08 0.47 0.53 0.6 0.4 0.48 ± 0.06 0.47 0.53 0.5 0.5 0.47 ± 0.02 0.47 0.53 0.4 0.6 0.5 ± 0.07 0.47 0.53 0.3 0.7 0.49 ± 0.02 0.47 0.53 0.2 0.8 0.50 ± 0.04 0.47 0.53 0. 0.9 0.53 ± 0.03 0.47 0.53 -.0 0.50 ± 0.04 0.47 0.53.0-0.53 ± 0.07 0.47 0.53 0.9 0. 0.5 ± 0.06 0.47 0.53 0.8 0.2 0.49 ± 0.06 0.47 0.52 0.7 0.3 0.50 ± 0.05 0.48 0.52 8

Etry Mxture C /EC 50, C 2 /EC 50,2 P ± SE P -,d P,e 30 [C 6 H 5 CH 2 Py]Br ( ad [C 6 H 5 CH 2 mm]br (2 0.6 0.4 0.50 ± 0.05 0.48 0.52 0.5 0.5 0.50 ± 0.04 0.48 0.52 0.4 0.6 0.48 ± 0.08 0.48 0.52 0.3 0.7 0.49 ± 0.09 0.48 0.52 0.2 0.8 0.5 ± 0.07 0.49 0.5 0. 0.9 0.53 ± 0.09 0.49 0.5 -.0 0.50 ± 0.04 0.49 0.5.0-0.50 ± 0.04 0.47 0.52 0.9 0. 0.5 ± 0.03 0.48 0.52 0.8 0.2 0.52 ± 0.05 0.48 0.52 0.7 0.3 0.5 ± 0.08 0.48 0.52 0.6 0.4 0.48 ± 0.04 0.48 0.52 0.5 0.5 0.52 ± 0.04 0.48 0.52 0.4 0.6 0.47 ± 0.03 0.48 0.5 0.3 0.7 0.49 ± 0.05 0.49 0.5 0.2 0.8 0.48 ± 0.02 0.49 0.5 0. 0.9 0.48 ± 0.04 0.49 0.5 -.0 0.50 ± 0.04 0.49 0.5 a Cocetrato of compoud the mxture. b EC 50 of compoud. c Oberved mxture vablty. d See equato 3a the ma text. e See equato 3b the ma text. Table 4. Cocetrato addto volvg [C 4 Chol]Br (, [C 4 Py]Br (2 ad [C 4 mm]br (3. Cell cultured for 24 hour the abece of FBS. C /EC 50, C 2 /EC 50,2 C 3 /EC 50,3 P ± SE P - P.0 - - 0.45 ± 0.02 0.46 0.53 -.0-0.47 ± 0.0 0.45 0.55 - -.0 0.54 ± 0.03 0.46 0.53 0.5 0.5-0.53 ± 0.02 0.45 0.54 0.5-0.5 0.56 ± 0.0 0.46 0.53-0.5 0.5 0.49 ± 0.02 0.45 0.54 0.3 0.3 0.4 0.54 ± 0.06 0.46 0.54 0.4 0.3 0.3 0.55 ± 0.03 0.46 0.54 0.3 0.4 0.3 0.52 ± 0.05 0.46 0.54 9

3. K-value calculato for lthum bromde ad chole bromde Table 5. Cocetrato addto of LBr ad chole bromde to [C 2 mm]br ad [C 8 mm]br ad cell cultured for 24 hour the abece of FBS. Etry Mxture C /EC 50, C 2 /EC 50,2 P ± SE P - P LBr ( ad [C 2 mm]br (2 2 LBr ( ad [C 8 mm]br (2 3 [C Chol]Br ( ad [C 2 mm]br (2 4 [C Chol]Br ( ad [C 8 mm]br (2.0-0.50 ± 0.0 0.48 0.52 0.9 0. 0.57 ± 0.02 0.48 0.52 0.8 0.2 0.60 ± 0.03 0.48 0.52 0.7 0.3 0.63 ± 0.04 0.48 0.52 0.6 0.4 0.62 ± 0.04 0.48 0.52 0.5 0.5 0.64 ± 0.05 0.48 0.52 0.4 0.6 0.63 ± 0.05 0.48 0.52 0.3 0.7 0.63 ± 0.04 0.47 0.53 0.2 0.8 0.60 ± 0.07 0.47 0.53 0. 0.9 0.55 ± 0.02 0.47 0.53 -.0 0.50 ± 0.03 0.47 0.53.0-0.49 ± 0.0 0.49 0.5 0.9 0. 0.49 ± 0.04 0.49 0.5 0.8 0.2 0.52 ± 0.06 0.48 0.52 0.7 0.3 0.52 ± 0.03 0.48 0.52 0.6 0.4 0.53 ± 0.04 0.48 0.52 0.5 0.5 0.57 ± 0.08 0.48 0.52 0.4 0.6 0.64 ± 0.03 0.48 0.52 0.3 0.7 0.64 ± 0.04 0.47 0.52 0.2 0.8 0.63 ± 0.04 0.47 0.53 0. 0.9 0.58 ± 0.04 0.47 0.53 -.0 0.53 ± 0.02 0.47 0.53.0-0.50 ± 0.03 0.47 0.53 0.9 0. 0.65 ± 0.04 0.47 0.53 0.8 0.2 0.66 ± 0.06 0.47 0.53 0.7 0.3 0.72 ± 0.03 0.47 0.53 0.6 0.4 0.78 ± 0.08 0.47 0.53 0.5 0.5 0.74 ± 0.06 0.47 0.53 0.4 0.6 0.65 ± 0.0 0.47 0.53 0.3 0.7 0.65 ± 0.07 0.47 0.53 0.2 0.8 0.63 ± 0.08 0.47 0.53 0. 0.9 0.63 ± 0.07 0.47 0.53 -.0 0.50 ± 0.04 0.47 0.53.0-0.5 ± 0.03 0.48 0.52 0.9 0. 0.63 ± 0.06 0.47 0.52 0.8 0.2 0.66 ± 0.05 0.47 0.53 0.7 0.3 0.72 ± 0.04 0.47 0.53 0.6 0.4 0.75 ± 0.08 0.47 0.53 0.5 0.5 0.76 ± 0.04 0.47 0.53 0.4 0.6 0.6 ± 0.04 0.47 0.53 0.3 0.7 0.53 ± 0.03 0.47 0.53 0.2 0.8 0.55 ± 0.05 0.46 0.53 0. 0.9 0.54 ± 0.03 0.46 0.54 -.0 0.50 ± 0.02 0.46 0.54 0

Fgure. K-value plot for LBr ad chole bromde ([C Chol]Br whe hypotoxc quatte are added to [C 2 mm]br (A ad [C 8 mm]br (B..60.20 A B.60.20 A B K-Value 0.80 0.40 K-Value 0.80 0.40 0.00 0.00 0.60 2.38 4.77 9.54 LBr (mm 0.00 0.00.36 2.72 5.43 0.87 [C Chol]Br (mm

4. Chage of mtochodral membrae potetal ad reactve oxyge pece HeLa cell cultured wth [C 2 mm][bf 4 ]. Fgure 2. Chage of mtochodral membrae potetal of HeLa cell cultured wth or wthout [C 2 mm][bf 4 ] for 48 hour detected by Rhodame 23. A: egatve cotrol, utaed cell; B: potve cotrol, taed ormal cell; C: treatmet, taed cell cultured wth [C 2 mm][bf 4 ] (6.3 mm; D: combato of B ad C. 2

Fgure 3. Reactve oxyge pece (ROS producto of HeLa cell cultured wth or wthout [C 2 mm][bf 4 ] for 48 hour A: egatve cotrol, utaed cell; B: potve cotrol, taed ormal cell; C: treatmet, taed cell cultured wth [C 2 mm][bf 4 ] (6.3 mm; D: combato of B ad C. 3

5. Dervato of equato 3a ad 3b the ma text. Equato 3a ad 3b were derved from equato 2 referece 33 the ma paper. The relatohp betwee P ad R ca be decrbed through equato S. P = R S. A ρ ad b deote the ame term, ad ρ beg the mple average of the ρ for each chemcal ( ρ the mxture, equato 2 from Rder ad LeBlac ca be wrtte a ρ ' = = P = R = ( = C EC 50, ρ ' = ( C EC = 50, ρ = = ( C EC = 50, b = S.2 If the cofdece terval of EC 50 ad b (at 95% cofdece level are take to accout, we get P = ( = EC 50, C.96SE ± EC 50, =.96SE b ± where SE deote tadard error ad the ample ze for compoud. Sce the cocetrato were choe o that C =, S.4 = EC50, the followg relatohp are oberved for SE EC50, >0 ad SE b, >0 C ( < S.5 =.96SEEC 50, EC50, ad ( C.96SE = EC50, EC50, o that >.96SEb,.96SEb, b b = = C C ( < ( SE S.7 =.96 EC = SE 50,.96 EC50, EC50, EC50, ad b, S.3 S.6 4

.96SE b,.96seb, b b = = C C ( > ( SE S.8 =.96 EC = SE 50.96, EC 50, EC50, EC50, The the upper ad lower lmt of the cofdece terval of P (P deotg the upper lmt, ad P - deotg the lower lmt wll be P = S.9.96SEb, b = C ( =.96SEEC50, EC50, = C.96SE ad P = S.0 ( EC 50, EC50,.96SEb, b = Equato S.9 ad S.0 are detcal to equato 3a ad 3b the ma text. 5