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Supplementary materials - Cancer Chemotherapy and Pharmacology
Computed determination of the in vitro optimal chemocombinations of
sphaeropsidin A with chemotherapeutic agents to combat melanomas
Aude Ingels1, Carina Dinhof2,3, Abhishek D. Garg4 , Lucia Maddau5, Marco Masi6, Antonio
Evidente6, Walter Berger 2,3 , Bieke Dejaegher7 and Véronique Mathieu1
1
Laboratoire de Cancérologie et Toxicologie Expérimentale, Université Libre de Bruxelles,
Boulevard du Triomphe, Accès 2, 1050 Ixelles, Belgium.
2
Department of Medicine I, Institute of Cancer Research, Medical University Vienna,
Spitalgasse 23, 1090 Vienna, Austria
3
Comprehensive Cancer Center, Medical University Vienna, Spitalgasse 23, 1090 Vienna,
Austria
4
Laboratory for Cell Death Research and Therapy (CDRT), Department of Cellular and
Molecular Medicine, KU Leuven University, Leuven, Belgium.
5
Dipartimento di Agraria, Sezione di Patologia vegetale ed Entomologia, Università degli
Studi di Sassari, Viale Italia 39, 07100, Sassari, Italy.
6
Dipartimento di Scienze Chimiche, Universita’ di Napoli Federico II, Complesso
Universitario Monte S. Angelo, Via Cintia 4, 80126 Napoli, Italy.
7
Laboratoire d’Analyse Instrumentale et de Bioélectrochimie, Université Libre de Bruxelles,
Boulevard du Triomphe, Accès 2, 1050 Ixelles, Belgium
Correspondence to:
Véronique Mathieu, MD, PhD
Laboratoire de Cancérologie et Toxicologie Expérimentale – Faculté de Pharmacie –
Université Libre de Bruxelles (ULB)
Campus de la Plaine – Boulevard du Triomphe – 1050 Brussels – Belgium.
Tel: +32 478 317 388 E-mail: [email protected]
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Supplementary materials - Cancer Chemotherapy and Pharmacology
Concentrations (µM)
Cisplatin
Sph A
0
0
N°Exp
1-3
Sph A
0
TMZ
0
4-6
6
0
6
0
7-9
0
100
0
1000
10-12
6
100
6
1000
13-15
0
50
0
500
16-18
6
50
6
500
19-21
3
0
3
0
22-24
3
100
3
1000
25-30
3
50
3
500
31
2
35
2
350
32
4
40
4
400
33
3
75
3
750
Table 1: Factorial experimental design performed to establish the model for
sphaeropsidin A (Sph A)/ cisplatin and Sph A/ temozolomide (TMZ) combinations and
tested on each cell line. 9 combinations of concentrations (µM) including the central point
and 3 test points permit to build the model.
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Supplementary materials - Cancer Chemotherapy and Pharmacology
A
B
C
Mean
B16F10
ln(B16F10)
16.045
2.021
Standard
Deviation
27.592
1.085
Minimum
Maximum
Centrum
2.076
0.731
100.000
4.605
51.038
2.668
Figs 1: Example of dissymmetric distribution of B16F10 dataset for sphaeropsidin A and
cisplatin. Boxplot for B16F10 data before (A) and after logarithm transformation (B). Table
C includes all absolute data visualized above.
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Supplementary materials - Cancer Chemotherapy and Pharmacology
Sph A and cisplatin
Sph A and temozolomide
Sph A and cisplatin
Sph A and temozolomide
Pre-treatment
Y = 0.618 + 0.201 * X1 + 0.149 * X2 - 0.125 * (X1*X1) 0.135 * (X2*X2) - 0.038 * (X1*X2)
Y= 0.319 + 0.251 * X1 + 0.096 * X2 - 0.010 * (X1*X1) 0.009 * (X2*X2) + 0.031 * (X1*X2)
Co-treatment
Y= 0.740 + 0.148 * X1 + 0.189 * X2 - 0.106 * (X1*X1) 0.156 * (X2*X2) - 0.105 * (X1*X2)
Y= 0.631 + 0.252 * X1 + 0.093 * X2 - 0.212 * (X1*X1) 0.055 * (X2*X2) - 0.033 * (X1*X2)
Table 2: The second-order polynomial equations built from experimental data to predict
relationship between concentrations of each drug and their combined response. X1 and
X2 are the coded values for [sphaeropsidin A (Sph A)] and [Cisplatin/Temozolomide], b0 is
the intercept coefficient, b1/ b2 are the linear coefficients, b1-1/ b2-2 are the quadratic
coefficients, and b1-2 is the factor interaction coefficient.
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Supplementary materials - Cancer Chemotherapy and Pharmacology
Pre-treatment
Cisplatin Y optimal
75
0.7247
70
0.7245
80
0.7223
65
0.7215
70
0.7172
85
0.7172
75
0.7162
60
0.7159
65
0.7155
80
0.7126
B
Co-treatment
Sph A Cisplatin Y optimal
1
4
75
0.8149
2
5
70
0.8139
3
4
70
0.8138
4
5
65
0.8131
5
4
80
0.8128
6
5
75
0.8114
7
4
65
0.8096
8
5
60
0.8092
9
4
85
0.8076
10
5
80
0.8058
Pre-treatment
Sph A
TMZ
Y optimal
1
6
1000
0.68
2
6
950
0.67
3
6
900
0.66
4
6
850
0.65
5
6
800
0.63
6
6
750
0.62
7
6
700
0.61
8
6
650
0.60
9
5
1000
0.59
10
6
600
0.59
D
Co-treatment
TMZ
Y optimal
850
0.73
800
0.73
900
0.73
750
0.73
950
0.72
700
0.72
850
0.72
900
0.72
1000
0.72
650
0.72
A
Sph A
1
5
2
5
3
5
4
5
5
6
6
5
7
6
8
5
9
6
10
6
C
Sph A
1
5
2
5
3
5
4
5
5
5
6
5
7
4
8
4
9
5
10
5
Tables 3: Summary tables of the ten best combinations of compounds (in µM) ranked
according to the best predicted response (Y optimal). Combinations of sphaeropsidin A
(Sph A) with cisplatin in pre-treatment and co-treatment conditions are presented in A and B
respectively. Combinations of Sph A with temozolomide (TMZ) in pre-treatment and cotreatment conditions are presented in C and D respectively.
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Supplementary materials - Cancer Chemotherapy and Pharmacology
Co-treatment with Sph A
Pre-treatment with Sph A
B
Cisplatin
A
D
Temozolomide
C
Figs 2: 2D contour plots obtained showing the isobole lines as a function of the levels of
two factors: sphaeropsidin A (Sph A) and cisplatin (A-B) or temozolomide (C-D)
concentrations. Two modes of sensitization are presented: pre-treatment (A-C) and cotreatment (B-D).
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