Supporting Information. Designing porphyrinic covalent organic frameworks for the photodynamic inactivation of bacteria Řež, Czech Republic

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1 Supporting Information Designing porphyrinic covalent organic frameworks for the photodynamic inactivation of bacteria Jan Hynek, a,b Jaroslav Zelenka, c Jiří Rathouský, d Pavel Kubát, d Tomáš Ruml, c Jan Demel, a and Kamil Lang a * a Institute of Inorganic Chemistry of the Czech Academy of Sciences, Husinec-Řež 11, Řež, Czech Republic b Department of Inorganic Chemistry, Faculty of Science, Charles University, Hlavova 23, Praha 2, Czech Republic c Department of Biochemistry and Microbiology, University of Chemistry and Technology Prague, Technická 5, Praha 6, Czech Republic d J. Heyrovský Institute of Physical Chemistry of the Czech Academy of Sciences, Dolejškova 3, Praha 8, Czech Republic *Corresponding author, lang@iic.cas.cz S-1

2 Content Syntheses of precursors Figure S1. 1 H NMR and 13 C NMR spectra of porphyrin 1. Figure S2. Mass spectra of 1. Figure S3. 1 H NMR and 13 C NMR spectra of porphyrin 2. Figure S4. Mass spectra of 2. Figure S5. Powder X-ray diffraction patterns of COFs. Figure S6. 13 C CPMAS-NMR spectra of COFs. Figure S7. FTIR of COFs. Figure S8. TGA/DTA curves of COFs. Figure S9. Nitrogen adsorption isotherms at 77K. Figure S1. Normalized fluorescence emission spectra in acetonitrile. Figure S11. Transient absorption decay curves of the triplet states in acetonitrile. Figure S12. Kinetics of O 2 ( 1 g ) luminescence generated by COFs in oxygen-saturated acetonitrile. Figure S13. Reaction of O 2 ( 1 g ) produced by 3D-PdTPP with 9,1-diphenylanthracene in acetonitrile. Figure S14. Microscopic fields with P. aeruginosa inspected by a confocal microscope after fixation and staining with Hoechst. Figure S15. Antimicrobial function of the COF coatings under a halogen lamp. Figure S16. Function of the 3D-TPP antibacterial coating irradiated with 46 nm light, 1 mw cm -2. S-2

3 Syntheses of precursors Preparation of 5,15-bis(4-formylphenyl)-1,2-diphenylporphyrin (1). A Schlenk tube was charged with 14 mg (225 μmol) of 5,15-dibromo-1,2-diphenylporphyrin, 75 mg (.5 mmol) of 4-formylbenzeneboronic acid, and 3.5 ml of.4m K 2 CO 3 (1.4 mmol). The tube was three times evacuated and purged with argon. The solution of 25 mg of Pd(PPh 3 ) 4 (22 μmol) in 6 ml of 1,4-dioxane was added. The reaction mixture was treated with 3 freezepump-thaw cycles. The solution was heated up to 1 C for 24 h. After cooling down, the solvent was evaporated, the remaining solid was dissolved in CH 2 Cl 2, and the mixture was washed with saturated water solution of NaCl and NaHCO 3. The organic fraction was dried over MgSO 4. The drying agent was filtered off, the solvent was evaporated, and the mixture was chromatographed using silica with CH 2 Cl 2 as an eluent. Yield: 125 mg, 83 %. 1 H NMR ( MHz, CDCl 3 ): δ (s, 2H); (m, 6 H); 8.22 (d, 3 J HH = 6.4 Hz, 4H); 8.28 (d, 3 J HH = 7.8 Hz, 4H); 8.4 (d, 3 J HH = 7.8 Hz, 4H); 8.8 (d, 3 J HH = 4.6 Hz, 4H); 8.9 (d, 3 J HH = 4.6 Hz, 4H; 1.38 (s, 2H) (Figure S1). 13 C NMR (1.59 MHz, CDCl 3 ): δ 118.6; 12.8; 126.8; 127.9; 134.5; 135.1; 135.6; 141.8; 148.5; (Figure S1). HRMS: calculated for C 46 H 31 O 2 N , found (Figure S2). Preparation of Pd(II)-5,15-bis(4-formylphenyl)-1,2-diphenylporphyrin (2). A roundbottom flask was charged with 85 mg (.13 mmol) of 1 and 135 mg (.45 mmol) of Pd(acac) 2, which were dissolved in 2 ml of toluene. The mixture was refluxed at 11 C for 48 h. The solvent was evaporated, the solid was dissolved in CH 2 Cl 2, and the solution was filtered. The filtrate was concentrated and chromatographed on a silica column with CH 2 Cl 2 as an eluent. After evaporation, the crude product was washed with MeOH several times to remove the excess of Pd(acac) 2. The progress of metalation was checked by luminescence spectroscopy. Yield: 79 mg, 8 %. 1 H NMR (6.17 MHz, CDCl 3 ): δ (m, 6 H); 8.15 (d, 3 J HH = 7.2 Hz, 4H); 8.26 (d, 3 J HH = 8. Hz, 4H); 8.34 (d, 3 J HH = 8. Hz, 4H); 8.74 (d, 3 J HH = 4.8 Hz, 4H); 8.84 (d, 3 J HH = 4.8 Hz, 4H); 1.36 (s, 2H) (Figure S3). 13 C NMR (15.91 MHz, CDCl 3 ): δ 12.4; 122.5; 126.9; 128.1; 13.7; 131.8; 134.2; 134.8; 135.8; 141.; 141.5; 141.9; 148.2; (Figure S3). HRMS: calculated for C 46 H 29 O 2 N 4 Pd , found (Figure S4). S-3

4 Figure S1. 1 H NMR (top) and 13 C NMR (bottom) spectra of porphyrin 1 in CDCl 3 measured on a Varian Mercury 4Plus Instrument. The chemical shifts are referred to the residual signals of CDCl 3 and are given in ppm. S-4

5 Figure S2. Electrospray ionization-mass spectrum of 1 in the positive mode (top) and highresolution mass spectrum (bottom). S-5

6 Figure S3. 1 H NMR (top) and 13 C NMR (bottom) spectra of porphyrin 2 in CDCl 3 measured on a JEOL 6 MHz NMR spectrometer. The chemical shifts are referred to the residual signals of CDCl 3 and are given in ppm. S-6

7 Figure S4. Electrospray ionization-mass spectrum of 2 in the positive mode (top) and highresolution mass spectrum (bottom). S-7

8 Intensity / a.u. Figure S5. Powder X-ray diffraction patterns of COFs. 3D-TPP 3D-PdTPP 2D-TPP Chemical shift / ppm Figure S6. 13 C CPMAS-NMR spectra of COFs. The spectra were recorded using a JEOL 6 MHz NMR spectrometer at 1 khz MAS rate; contact time was set to 6 ms and relaxation delay was 5 s. S-8

9 Transmittance Transmittance 3D-TPP b 3D-PdTPP b 2D-TPP b Wavenumber / cm -1 3D-TPP 3D-PdTPP a 2D-TPP a a Wavenumber / cm -1 Figure S7. FTIR of COFs: region 4-2 cm 1 was measured in KBr pellets and the region 2-4 cm 1 was measured with a diffuse reflection accessory; residual amino (a) and aldehyde (b) groups are labeled. S-9

10 512 C 641 C --> exo 154 C 54 C 639 C 387 C <+> <+> 35 C 41 C --> exo 11 C <+> m / % 352 C 41 C <+> <+> 512 C 518 C 67 C --> exo <+> 597 C 158 C 4 C 59 C TG A -3.5% (3-313 C) -89.3% ( C) DTG / (mg min -1 ) DTA / µv Temperature / C m / % TG DTG / (mg min -1 ) DTA / µv % (3-316 C) -75.3% ( C) -2 B Temperature / C m /% TG C -6.1% (3-338 C) DTG / (mg min -1 ) DTA / µv -89.3% ( C) Temperature / C Figure S8. TGA/DTA curves of 3D-TPP (A), 3D-PdTPP (B), and 2D-TPP (C). The measurements were performed in synthetic air atmosphere with a flow rate of 3 ml min -1 and a temperature gradient of 5 C min -1. S-1

11 Intensity / a.u. Nitrogen adsorption / cm 3 g -1, STP 2 3D-TPP 3D-PdTPP 2D-TPP P/P Figure S9. Nitrogen adsorption isotherms at 77K for 2D-TPP (blue), 3D-TPP (black) and 3D- PdTPP (red). 1. c b a Wavelength / nm Figure S1. Normalized fluorescence emission spectra of 2D-TPP (a) and 3D-TPP (b) dispersed in acetonitrile compared with that of TPP (c). S-11

12 A Absorbance.2 Oxygen T = 5 ns Air T = 216 ns Argon T = 68 s Time / s Time s Time s Absorbance B Oxygen T = 55 ns Air T = 23 ns Argon T = 49 s Time s Time s Time / s Absorbance.2 C Oxygen T = 43 ns Air T = 19 ns Argon T = 46 s Time s Time / s Time / s S-12

13 D Absorbance.3.2 Absorbance Wavelength / nm Wavelength / nm Figure S11. Transient absorption decay curves of the triplet states of 5,1,15,2- tetraphenylporphyrin (TPP) (A), 3D-TPP (B), and 2D-TPP (C) in oxygen-, air-, or argonsaturated acetonitrile. The traces were recorded at 46 nm; the samples were excitated by a 425 nm laser pulse; red lines represent corresponding monoexponential fits. D) Left panel: Transient absorption spectrum of TPP in argon-saturated acetonitrile ( exc = 425 nm, 2 s after excitation). Right panel: Transient absorption spectrum of 3D-TPP in airsaturated acetonitrile ( exc = 38 nm,.7 s after excitation). S-13

14 c/c / % Intensity / a.u Intensity / a.u. 4 3 TPP 4 3 3D-TPP D-TPP D-PdTPP Time / s Time / s Figure S12. Kinetics of O 2 ( 1 g ) luminescence at 127 nm generated by TPP and COFs in oxygen-saturated acetonitrile upon excitation by a 425 nm laser pulse. The signals were recorded for the dispersions with matched absorbance at the excitation wavelength. Red lines represent corresponding monoexponential fits. 1 8 First run Second run Third run Time / min Figure S13. Reaction of photosensitized O 2 ( 1 g ) produced by 3D-PdTPP in acetonitrile with 9,1-diphenylanthracene: first, second, and third run. S-14

15 Figure S14. Microscopic fields with P. aeruginosa inspected by a confocal microscope at 1 magnitude after fixation and staining with Hoechst. Scale bar represents 1 μm. The Hoechst staining was measured using 45 nm laser excitation. A) Blank (polymer coating without 3D-TPP) in the dark; B) Blank after irradiation with 46 nm light for 24 h. C) Fluorescence of the 3D-TPP coating; D) Bacterial biofilm on the 3D-TPP coating in the dark; E) Merging of two panels on the left. F) Fluorescence of the 3D-TPP coating after irradiation with 46 nm light for 24 h; G) Bacterial biofilm on the 3D-TPP coating after irradiation with 46 nm light for 24 h; H) Merging of two panels on the left. S-15

16 Figure S15. Antimicrobial function of the COF coatings on the density of P. aeruginosa (A) and E. faecalis (B) biofilms: 24 h incubation in the dark (black) or under a halogen lamp (red). The blank experiments were performed with the polymer coating without COF. In all cases, the amount of the biofilm is quantified as percentage of the surface covered with bacteria (y axis). The experiments were analyzed by the Student t-test and the results with p <.1 (labeled as *) are considered as significant. S-16

17 Figure S16. 3D-TPP antibacterial coating vs. blank coating, 46 nm light, 1 mw cm -2, biofilm of P. aeruginosa: A) Inhibition of biofilms under 46 nm light, 24 h. B) Biofilms were formed during 24 h incubation in the dark, followed by 4 h irradiation with 46 nm light. The experiments were analyzed by the Student t-test and the results with p <.1 (labeled as *) are considered as significant. S-17