Synergistic Combination Chemotherapy of Camptothecin and Floxuridine through Self-assembly of Amphiphilic Drug-Drug Conjugate Minxi Hu, Ping Huang, Yao Wang, Yue Su, Linzhu Zhou, Xinyuan Zhu,* and Deyue Yan* School of Chemistry and Chemical Engineering, Shanghai Key Lab of Electrical Insulation and Thermal Aging, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, P. R. China * Correspondence to: [email protected]; [email protected]. Fax: +86-21-54741297 Table of Contents Supplementary Figures 1 Figure S1: H NMR spectra of CPT and CPT-COOH in DMSO-d6. Figure S2: FTIR spectra of CPT, FUDR, and CPT-FUDR conjugate. Figure S3: (A) UV/Vis spectra of CPT, FUDR and CPT-FUDR conjugate in acetonitrile. (B) Fluorescence emission spectra of CPT (λ ex = 360 nm, λ em = 430 nm) and CPT-FUDR conjugate (λ ex = 360 nm, λ em = 426 nm) in DMSO. Figure S4: The UV absorbance of DPH in an aqueous solution of CPT-FUDR nanoparticles with various concentrations. The CAC value of CPT-FUDR nanoparticles is about 15 µM. Figure S5: Influence of storage on (A) diameter (the diameter measured in Day 1 was set as control, and the data are presented as ratios to the control) and (B) PDI of CPT-FUDR nanoparticles with the extension of time in water. Figure S6: (A)Total ion chromatography (TIC) of CPT and CPT-FUDR . (B, C) Extracted ion chromatography (EIC) of CPT-FUDR (m/z = 693.1835, (M+H +)) and CPT (m/z = 349.1165, (M+H +)). The retention time of CPT-FUDR and CPT is 3.92 and 3.79 min, respectively. Figure S7: (A) Total ion chromatography (TIC) of the cell extracts. (B, C) Extracted ion chromatography (EIC) of CPT-FUDR (m/z = 693.1853, (M+H +)) and FUDR (m/z = 349.1205, (M+H +)). The retention time of CPT-FUDR and CPT is 3.93, and 3.80 min, respectively. Figure S8: Flow cytometry histogram profiles of HT-29 cells treated with CPT-FUDR nanoparticles containing core-encapsulated Nile red for 30 min, 2 h and 4 h. 1 Figure S1. H NMR spectra of CPT and CPT-COOH in DMSO-d6. Figure S2. FTIR spectra of CPT, FUDR, and CPT-FUDR conjugate. Figure S3 . (A) UV/Vis spectra of CPT, FUDR and CPT-FUDR conjugate in acetonitrile. (B) Fluorescence emission spectra of CPT (λ ex = 360 nm, λ em = 430 nm) and CPT-FUDR conjugate (λ ex = 360 nm, λ em = 426 nm) in DMSO. Figure S4. The UV absorbance of DPH in an aqueous solution of CPT-FUDR nanoparticles with various concentrations. The CAC value of CPT-FUDR nanoparticles is about 15 µM. Figure S5. Influence of storage on (A) diameter (the diameter measured in Day 1 was set as control, and the data are presented as ratios to the control) and (B) PDI of CPT-FUDR nanoparticles with the extension of time in water. Figure S6. (A)Total ion chromatography (TIC) of CPT and CPT-FUDR. (B, C) Extracted ion chromatography (EIC) of CPT-FUDR (m/z = 693.1835, (M+H +)) and CPT (m/z = 349.1165, (M+H +)). The retention time of CPT-FUDR and CPT is 3.92 and 3.79 min, respectively. Figure S7. (A) Total ion chromatography (TIC) of the cell extracts. (B, C) Extracted ion chromatography (EIC) of CPT-FUDR (m/z = 693.1853, (M+H +)) and FUDR (m/z = 349.1205, (M+H +)). The retention time of CPT-FUDR and CPT is 3.93, and 3.80 min, respectively. Figure S8. Flow cytometry histogram profiles of HT-29 cells treated with CPT-FUDR nanoparticles containing core-encapsulated Nile red for 30 min, 2 h and 4 h. .