Electronic Supplementary Material (ESI) for Nanoscale. This journal is © The Royal Society of Chemistry 2021 Supporting information for Novel multifunctional nano-hybrid polyhedral oligomeric silsesquioxane-based molecules with high cell permeability: Molecular design and application for diagnosis and treatment of tumors Gang Wei1, Kezhen Zhang2, Yuanlong Gu3*, Shanyi Guang2 *, Jihong Feng 4, Hongyao Xu1* Dr. G. Wei，Prof. H. Y. Xu, 1State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, Research Center for Analysis and Measurement & College of Materials Science and Engineering, Donghua University, Shanghai 201620, China E-mail:[email protected] Prof. S. Y. Guang 2School of Chemistry, and Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620, China E-mail: [email protected] Dr. Y. L. Gu 3Department of Oncology, Taizhou Municipal Hospital, the Affiliated Hospital of Medical College of Taizhou University, Taizhou, Zhejiang, 318000, P. R. China E-mail: [email protected] Dr. J. H. Feng 4Department of oncology, Lishui People's Hospital, Sixth Affiliated Hospital of Wenzhou Medical University, Lishui, Zhejiang 323000, China 1 Targeting monomers (R1) and fluorescent monomers (R2) have been prepared according to the reported literature. [1,2] 1. Synthesis of acryl folic acid (R1) 8.828 g (0.01 mol) of folic acid was dissolved in 50 mL of dried DMSO with vigorous stirring at 30℃ to form a uniform solution under dark, then 6.0 mL triethylamine as acid binding agent was injected. In addition, 2.0 mL (0.024 mol) of acryloyl chloride in 20 mL DMSO was dropwise added into the above dissolution for 30 min. The reaction solution maintained at 30 °C for 24 h. The reaction solution was extracted with acetone and then three times with ethanol. Yield: 4.42 g (50 %) 1HNMR (600 MHz, CDCl3, 298 K, δ/ppm): δ 8.64 (s, 1 H), 7.96 (s, 1 H), 7.63 (s, 1 H), 6.93 (s, 1 H), 6.66 (s, 1 H), 4.49 (s, 1 H), 4.28 (s, 1 H), 2.27 (s, 1 H), 2.01 (s, 1 H). FTIR (KBr): v=3000, 1635 cm−1 (NeC), 1243 cm−1 (CeO), 1618 cm−1 (C]C), 1591, 1580, 1496, 1454 cm−1 (C6H6). H H2N N N O H N N H2C C C Cl N H H O N COOH Rt , 24h O COOH O H H H2C C C N N N H H N N N H O N COOH O COOH 2. Synthesis of fluorescent monomers (R2) 3.00 g (9.0×10−3 mol) fluorescein was dispersed in 20 mL dichloromethane. After dissolution, 3.0 mL triethylamine was added in the above solution. The mixture solution was kept in ice bath environment to obtain cool and uniformity mixture solution with magnetic stirring, then acryloyl chloride (0.90 mL, 0.011 mol) dissolved in 20 mL dichloromethane was slowly dropwise added for 30 min in ice baths. The mixture solution was kept at room temperature (25℃) for 24 h. After rotational evaporation, orange crystal raw product was obtained, which was further purified by 2 column chromatography machine with ethanol and chloroform (the volume ratio 1:30) as eluent. At last, allyl fluorescein with light yellow was obtained. Yield: 2.35 g (78.3 %) 1HNMR (600 MHz, CDCl3, 298 K, δ/ppm): δ 8.05 (d, 1 H), 7.68 (d, 1 H), 7.64 (d, 1 H), 7.29 (s, 1 H), 7.14 (s, 1 H), 6.82 (s, 1 H), 6.71 (s, 1 H), 6.65 (s, 1 H), 6.62 (s, 1 H), 6.55 (s, 1 H), 6.35 (s, 1 H), 6.10 (s, 1 H). FTIR (KBr): v =1768 cm−1 (C=O), 1744 cm−1 (C=O), 1618 cm−1 (C=C), 1591, 1580, 1496, 1454 cm−1 (C6H6), 1243 cm−1 (CeO), 1111 cm−1 (CeOeC). O O HO O OH H C C C O O OH H2C C C Cl 2 H H COOH Rt , 24h HOOC 3 Figure S1 The FTIR spectrum of R1 1 Figure S2 The H NMR of R1 Figure S3 The FTIR spectrum of R2 4 1 Figure S4 The H NMR spectrum of R2 Figure S5 The FTIR spectrum of R4 1 Figure S6 The H NMR of R4 5 Figure S7 The 1H NMR of PEG-Ac Figure S8 The FTIR spectrum of PEG-Ac Figure S9 29Si NMR of eight mercapto POSS 6 Figure S10 The 1H NMR of POSS Figure S11 The FTIR spectrum of POSS SH PPFFGH PPFFG PPFF PPF 4000 3500 3000 2500 2000 1500 1000 500 Wavenumber/cm-1 Figure S12 The FTIR spectrum of multifunctional molecules synthesis process 7 ) u . 700 (a) Ex Em a ( y t 600 i s n 500 e t n I 400 e c 300 n e c 200 s e r 100 o u l F 0 400 450 500 550 600 650 Wavelength / nm Figure S13 Excitation and emission spectra of H1 in Ethanol/H2O (1/1, v/v) ( λex =430 nm), ) 800 u 700 . ) u a . a 600 ( ( 700 y t i y 500 s t n i e t s 400 n I 600 n e c 300 e n e t c s 200 n 500 e I r o u 100 l e F c 400 0 n 0 20 40 60 80 100 e -6 c [PPFGH] 10 M s 300 e r o 200 u l F 100 0 500 550 600 650 700 Wavelength / nm Figure S14 The fluorescence emission spectra of H3 with different concentrations (0-100 µM) in EtOH/H2O (v/v, 1/1, λex=440 nm), Figure S15 The fluorescence emission spectra of H3 with different concentrations (0-100 µM) in EtOH/H2O (v/v, 1/1, λex=440 nm), 8 Figure S16 the MS of H1 Figure S17 the MS of H2 9 Figure S18 the MS of H3 References [1] S. Y. Guang, J. C. Tian, G. Wei, Z. Q. Yan, H. F. Pan, J. H. Feng, H. Y. Xu, talanta, 2017, 170, 89-96. [2] G. Wei, G. Zhao, N. B. Lin, S. Y. Guang, H.Y. Xu, Colloid. Surface. A, 2020, 599, 124863-124870. 10.