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Supplementary Information Section

Meso-mono-[4-(1,4,7-triazacyclononanyl)]-tri(phenyl)]porphyrin and respective

zinc(II)-complex: complete characterization and biomolecules binding abilities

Bruna L. Auras,a Vanessa A. Oliveira,c Hernán Terenzic, Ademir Nevesa and Bernardo A. Iglesias*a,b

aDepartamento de Química, Universidade Federal de Santa Catarina, Florianópolis, SC 88040-900, Brazil.

bDepartamento de Química, Universidade Federal de Santa Maria, Santa Maria, RS 97105-900, Brazil.

cCentro de Biologia Molecular Estrutural, Universidade Federal de Santa Catarina, Florianópolis, SC

88040-900, Brazil.

Corresponding Author e-mail address: B.A. Iglesias (bernardopgq@gmail.com)

Contents

1. Experimental details…………………………………………………………………….………..……………….………… 2

1.1. Synthesis of free-base porphyrin 4………………………………………………………………….……….………….. 2

1.2. Synthesis of zinc(II)-porphyrin 5……………………………………………………………………….…………………. 3

2. Spectral profile…………………………………………………………………………………….……….…………………... 4

2.1. ESI-MS and HRMS-ESI analysis………………………………………………………………………………….………….. 4

2.2. 1H and 2D COSY NMR spectra………………………………………………………………….…………..…….………… 7

2.3. FT-IR analysis……………………………………………………………………..……….………………………….…………… 10

2.4. Electrochemical analysis……………………………………………………………….…………………………………….. 10

2.5. Spectroelectrochemistry data………………………………………………………………………………….……..……. 12

2.6. Photostability assay…………………………………………………………………………………………………..…..…….. 13

2.7. UV-vis and emission ct-DNA titration data………………………………………………….………..…..…………. 14

2.8. References………………………………………………………………………………………………………………..………… 16

Electronic Supplementary Material (ESI) for Photochemical & Photobiological Sciences.This journal is © The Royal Society of Chemistry and Owner Societies 2016

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1. Experimental details

1.1. Synthesis of meso-mono-[4-(1,4,7-triazacyclononanyl)]-tri(phenyl)]porphyrin 41

To a solution of 5-(4-aminophenyl)-10,15,20-triphenylporphyrin 12 (0.05 g; 0.079

mmol; 1.0 equiv.) and Cmff-TACN 2 (0.043 g; 0.120 mmol; 1.51 equiv.) in toluene (10.0

mL), La(OTf)3 (0.009 g; 0.016 mmol; 4.9 equiv.) was added at room temperature. The

reaction mixture was stirred under reflux for 10 h. The reaction mixture was allowed to

cool at 25 ºC and filtered through a celite plug. The filtrate was evaporated under

reduce pressure to produce a purple solid corresponding to the iminoporphyrin 3. To a

solution of iminoporphyrin 3 in chloroform (10.0 mL), a solution of NaBH4 (0.015 g;

0.190 mmol; 2.4 equiv.) in methanol (1.0 mL) was added, and the reaction mixture was

stirred at room temperature for 20 min. After completation of the reaction, the

mixture was added with water and organic layer was dried over anhydrous Na2SO4.

The organic layer was evaporated under reduced pressure. Thus, the crude product

obtained was purified over silica gel column using chloroform/methanol mixture (4:1)

as eluent. Porphyrin 4 was recrystallizated with chloroform/methanol mixture (1:1)

affords a sufficient pure solid.

Porphyrin 4: Purple solid; yield: 44 %; M.p. > 300ºC (decomp.); UV-vis max (CH2Cl2; in

M-1 cm-1): 418 (234.422), 516 (16.218), 553 (10.715), 592 (8.128) and 647 (7.943); em

(CH2Cl2; exc = 420 nm): 652 and 720 nm; IR (max; cm-1): 3500 (external NH), 3022 (NH

porphyrin), 1558 (C-H) and 820 (C-N); 1H NMR (400 MHz, CDCl3) in ppm: -2,76 (bs,

NH, 2H); 8,93 (d, J = 4,0 Hz, -H, 2H); 8,82 (bs, -H, 6H); 8,22 (d, J = 8,0 Hz, o-HPh, 6H);

8,02 (m, m-HPh, 2H); 7,96 (s, H-Ph Cmff, 1H); 7,76 (m, p- e m-HPh, 9H); 7,10 (m, o-HPh,

2H); 7,00 (s, H-Ph Cmff, 1H); 4,59 (s, CH2-NH-Ar, 1H); 3,86 (s, CH2-Ar, 2H); 2,95 - 2,31

(m, CH3-Ar e CH2-TACN, 17H); 1,25 (s, CH3isoprop-TACN, 12H); HRMS-ESI: m/z =

3

calculated 974.5354, found 974.5474 [M]+; Anal. Calc. for C65H66N8O•2H2O: C, 77.20; H,

6.98; N, 11.08; found: C, 77.06; H, 7.06; N, 11.04.

1.2. Synthesis of meso-mono-[4-(1,4,7-triazacyclononanyl)]-

tri(phenyl)]porphyrinate zinc(II) 5

To a solution of the corresponding free-base porphyrin 4 (0.025 g; 0.0395 mmol; 1.0

equiv.) in chloroform (5.0 mL), a solution of zinc(II) acetate dehydrate (0.026 g; 0.118

mmol; 3.0 equiv.) in methanol (2.0 mL) was added. The reaction mixture was stirred at

room temperature for 1 h. After complexation of the free-base porphyrin, the mixture

was washed with water, and the CHCl3 layer was dried over anhydrous Na2SO4. The

organic layer was evaporated under reduced pressure to afford the crude product,

which was purified by column chromatography in silica gel using chloroform as eluent,

following by recrystallization in chloroform/methanol (1:1) mixture.

Porphyrin 5: Purple solid; yield: 90 %; M.p. > 300ºC (decomp.); UV-vis max (CH2Cl2; in

M-1 cm-1): 425 (266.677), 558 (23.337) and 600 (17.477); em (CH2Cl2; exc = 420 nm):

602 and 650 nm; IR (max; cm-1): 3488 (external NH), 1502 (C-H) and 814 (C-N); 1H NMR

(400 MHz, CDCl3) in ppm: 8,90 (d, J = 4,0 Hz, -H, 2H); 8,79 (bs, -H, 6H); 8,21 (d, J =

8,0 Hz, o-HPh, 6H); 7,99 (m, m-HPh, 2H); 7,95 (s, H-Ph Cmff, 1H); 7,72 (m, p- e m-HPh,

9H); 7,08 (m, o-HPh, 2H); 7,01 (s, H-Ph Cmff, 1H); 4,57 (s, CH2-NH-Ar, 1H); 3,83 (s, CH2-

Ar, 2H); 2,92 - 2,29 (m, CH3-Ar e CH2-TACN, 17H); 1,23 (s, CH3isoprop-TACN, 12H); HRMS-

ESI: m/z = calculated 1037.4567, found 1037.4568 [M+H]+; Anal. Calc. for

C65H64N8OZn•7H2O: C, 67.03; H, 6.75; N, 9.62; found: C, 66.31; H, 6.24; N, 9.44.

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2. Spectral Profile

2.1. ESI-MS and HRMS-ESI analysis

Figure S1. ESI-(+)-MS and ESI-(+)-MS/MS spectrum of porphyrin 4.

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Figure S2. ESI-(+)-HRMS spectrum of porphyrin 4.

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Figure S3. ESI-(+)-HRMS spectrum of porphyrin 5.

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2.2. 1H and 2D COSY NMR spectra

Figure S4. 1H NMR spectrum of porphyrin 4 in CDCl3.

Figure S5. 1H NMR spectrum of porphyrin 5 in CDCl3.

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Figure S6. 1H-1H COSY 2D NMR spectrum of porphyrin 4 in CDCl3.

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Figure S7. 1H-1H COSY 2D NMR spectrum of porphyrin 5 in CDCl3.

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2.3. FT-IR analysis

Figure S8. IR analysis (KBr pellets) of porphyrin derivatives.

2.4. Electrochemical analysis

Figure S9. Cyclic voltammograms of Cmff-TACN 2 solution in DCM, 0.1 M TBAPF6, using glassy carbon

electrode, at scan rate 50 - 200 mV s-1.

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Figure S10. Cyclic voltammograms of porphyrin 4 solution in DCM, 0.1 M TBAPF6, using glassy carbon

electrode, at scan rate 50 - 200 mV s-1.

Figure S11. Cyclic voltammograms of porphyrin 5 solution in DCM, 0.1 M TBAPF6, using glassy carbon

electrode, at scan rate 50 - 200 mV s-1.

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2.5. Spectroelectrochemistry data

Figure S12. Spectroelectrochemistry of porphyrin 5 solution in DCM, 0.1 M TBAClO4, in the oxidation (a)

0.0 V to +1.10 V, (b) +1.10 V to +1.50 V and (c) +1.50 V to+ 1.95 V range. In the reduction (d) 0.0 V to -

1.20 V range, respectively.

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2.6. Photostability assay

Fig. S13 Photo-oxidation of DPA (50 μM) in DMF solution with or without porphyrin derivatives TPP,

ZnNH2TPP, 4 and 5 at 0.5 μM, after red light irradiation (LED array system) at a potency of 26 mW cm-2.

The maximum absorbance of DPA was recorded at 378 nm.

Table S1. Photostability of tris-HCl buffer solution of porphyrin derivatives, after irradiation with white

light (400-800 nm) at a fluence rate of 30.0 mW cm-2 for different periods of time (0 – 60 min).

Compounds Irradiation Time (min.) / %

0 10 20 30 40 50 60

TPP 100 96 92 90 84 84 75

ZnNH2TPP 100 86 76 63 62 59 55

Porphyrin 4 100 88 77 69 60 60 59

Porphyrin 5 100 89 89 81 74 66 64

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2.7. UV-vis and emission ct-DNA titration data

Figure S14. Electronic UV-vis absorption spectra of (a) TPP and (b) ZnNH2TPP, both [porphyrin] = 3.0

μM, monitoring the Soret band, with increasing ct-DNA concentrations ranging from 0.0 to 10.0 μM in

CH3CN-tris-HCl buffer mixture solution.

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Figure S15. Emission spectra of porphyrins (a) TPP and (b) ZnNH2TPP (1.0 – 2.0 μM range) with

increasing ct-DNA concentrations ranging from 0.0 to 10.0 μM CH3CN-tris-HCl buffer mixture solution

(λexc = 420 nm).

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Figure S16. Emission spectra of porphyrins (c) porphyrin 4 and (d) porphyrin 5 (1.0 – 2.0 μM range) with

increasing ct-DNA concentrations ranging from 0.0 to 10.0 μM CH3CN-tris-HCl buffer mixture solution

(λexc = 420 nm).

2.8. References

1 S. Sharma, N. Nath, Beilstein J. Org. Chem., 2013, 9, 496–502.

2 D. . alas e . Ma imo a . Rybic a- asin s a, A. Lipke, D. Gryko, J. Porphyrins

Phthalocyan., 2014, 18, 493–505.