CohortTreatment groups DoseMode of administration Number of animals DT01 + OXA + 5-FU sacrificed day 22 Vehicle (NaCl + glucose) 0.9% 5% IP 9 OXA 5-FU 6mg/kg 25mg/kg IP 10 DT015mgIP10 DT01 OXA 5-FU 2h 5mg 6mg/kg 25mg/kg IP 10 DT01 OXA 5-FU 4h 5mg 6mg/kg 25mg/kg IP 10 DT01 + OXA + 5-FU termination guidelines DT01 OXA 5-FU 4h 5mg 6mg/kg 25mg/kg IP 10 Table S1: Treatment group allocation of mice post intrahepatic grafting

Figure S1 Vehicle DT01 OXA DT01 + OXA Vehicle DT01 5-FU DT FU Figure S1: DT01 in combination with single agent chemotherapy Mean liver tumor volume (mm 3 ) after treatment with DT01 in association with a single chemotherapy agent (A) oxaliplatin (OXA) or (B) 5-fluorouracil (5-FU). AB

Score1234 Vehicle47% (7)53% (8)00 DT0150% (5) 00 CT40% (4)60% (6)00 DT01 + CT 2h25% (2)13% (1)62% (5)0 DT01 + CT 4h0038% (3)62% (5) Figure S2 DT CT h 4h Figure S2: Proportion of apoptotic nuclei in HES sections The degree of apoptosis in each treatment group was estimated by histological evaluation of HES sections from representative non-necrotic fields at high power. The degree of apoptosis was allocated scores between 1-4; where 1 represents weak (<5%), 2: moderate (5-10%), 3: significant (10-20%) and 4: highly significant (20-50%). For ease of assessment, scores 3 and 4 were combined in the histogram. In the histogram, weak, moderate and significant apoptosis are illustrated in white, grey or black bars, respectively.

Figure S3 Figure S3: Average body weight (g) of mice during treatment Body weight fluctuations in the HT29 CRC metastatic model treated with DT01 in association with (A) oxaliplatin and 5-fluorouracil and (B) doxorubicin. No significant fluctuations in weight were noted prior, during or after treatment in these mice.

NT DT01 DT01+OXA/5-FU ASTALTALP AMYTBIL Figure S4 Figure S4: Liver function assessment Blood samples were obtained through submandibular bleeding in lithium heparin tubes (Sarstedt) on days 0, 4 and 18 post treatment. Plasma alanine transaminase (ALT), aspartate aminotransferase (ASAT), alkaline phosphatase (ALP), glutamyl transpeptidase (GGT), amylase (AMYL) and total bilirubin (TBIL) were measured using an MS-Scan II.

A B Figure S5 Figure S5: Average body and liver weights (g) of mice during treatment with escalating doses of DT01 in association with OXA or 5-FU (A) Average body weight and (B) liver weight fluctuations in mice treated with two cycles of DT01 in association with oxalipaltin (OXA) or 5-fluorouracil (5-FU) in a NMRI NU/NU mouse model.

Treatment CRC liver metastasis model (HT29) Liver tumour volume (mm3) Necrosis (%) Proliferation Ki67 (%) Micro- vascularization CD31 (vessel count) Vehicle only 1086 ± ± 177 ± 238 ± 2 DT ± ± 280 ± 138 ± 2 CT 872 ± ± 280 ± 136 ± 2 DT01 + CT 2h 526 ± 8946 ± 1156 ± 1324 ± 1 DT01 + CT 4h501 ± 9386 ± 24 ± 18 ± 1 Table S2: Summary of main findings Note: Results are expressed as an average ± SEM.