Supporting information 1

Supporting information 1a. 1H-NMR and 13C-NMR spectra of 1-(Phenylsulfonyl)-1H-pyrrole (1).

Supporting information 1b. 1H-NMR and 13C-NMR spectra of 3-(tert-Butyl)-1-(phenylsulfonyl) -1H-pyrrole (2).

Supporting information 1c. 1H-NMR and 13C-NMR spectra of 4,4'-di-tert-butyl-1,1'- bis(phenylsulfonyl)-1H,1'H-2,2'-bipyrrole (3).

Supporting information 1d. 1H-NMR and 13C-NMR spectra of 4,4'-di-tert-butyl-1H,1'H-2,2'- bipyrrole (4).

Supporting information 1e. 1H-NMR and 13C-NMR spectra of 4,4'-di-tert-butyl-1H,1'H- [2,2'-bipyrrole]-5,5'-dicarbaldehyde (5).

Supporting information 1f. 1H-NMR and 13C-NMR spectra of 2,7,12,17-tetra-tert- butylporphycene (6).

Supporting information 1g. 1H-NMR, 13C-NMR and HRMS spectra of 7,12,17-Tri-tert-butylporphycene- 3-sulfonyl chloride (7, TBPoS-Cl).

Supplementary Figure 1h. 1H-NMR, 13C-NMR and HRMS spectra of 7,17-Di-tert- butylporphycene-3,13-disulfonyl chloride (8, TBPoS-2Cl).

Supplementary Figure 1i. 1H-NMR, 13C-NMR and HRMS spectra and HPLC analysis of 7,12,17-Tri-tert- butylporphycene-3-N-(2-(2-hydroxyethoxy) ethyl) sulfonamide (9, TBPoS-OH).

Supplementary Figure 1j. 1H-NMR, 13C-NMR and HRMS spectra and HPLC analysis of 7,17-Di-tert- butylporphycene-3,13-di-N-(2-(2-hydroxyethoxy) ethyl) sulfonamide (10, TBPoS-2OH).

Supporting information 2. The crystals of TBPoS-OH and TBPoS-2OH

Supplementary Figure 2. The properties of the crystals of TBPoS-OH and TBPoS-2OH. ORTEP drawings of TBPoS-OH (a) and TBPoS-2OH (b) with thermal ellipsoids at 50% probability. Top: perspective view. Bottom: macrocycle plane view. For clarity, only N atoms and selected C and S atoms have been numbered; hydrogen atoms have been omitted. Selected C-C and C-N bond distances and crystallographically determined internal cavity dimensions (in pm) of TBPoS-OH (c) and TBPoS-2OH (d). Perspective view of crystal packing in the lattice of TBPoS-OH (e) and TBPoS-2OH (f) viewing from b-axis.

Note: The X-ray data showed that the molecular structure of TBPoS-2OH is a nearly planar structure and comparable to normal porphycenes. The pyrrole-pyrrole dihedral angles are 1.74˚ (N1-C1-C20-N4), 1.63˚ (N1-C4-C7-N2), 0.67˚ (N2-C10-C11-N3), and 3.71˚ (N3-C14-C17-N4), respectively. In addition, the bond distances of C-C and C-N within the macrocycle plane are summarized in Supplementary Figure 2c. We emphasize that the two sulfonamide linkages of TBPoS-2OH significantly affected the dimensions of the N4 core, which leads to a more rectangular cavity than the H-tetra-substituted porphycene derivative [1]. The size of the cavity remains similar to that of the EtNBS [2].

On the other hand, the TBPoS-OH prepared with only one sulfonamide linkage distorted its inner N4 cavity significantly with the following pyrrole-pyrrole dihedral angles: 5.20˚ (N1-C1-C20-N2), 9.95˚ (N1-C4-C7-N4), 1.04˚ (N4-C10-C11-N3), and 4.84˚(N3-C14-C17-N2) as shown in Supplementary Figure 2a. The dimensions of the N4 core are summarized in Supplementary Figure 2d, which indicated that it is a distorted rectangular cavity due to the presence of the single high steric hindered sulfonamide linkage. This slight structural distortion in the inner N4 cavity between TBPoS-2OH and TBPoS-OH can also be reflected by their corresponding molecular packing within the lattice (Supplementary Figure 2e, f). We emphasize that TBPoS-2OH is a rather loosely face-to-face packed system within the lattice relative to the closely zig-zag packing found in the crystal lattice of TBPoS-OH.

References:

1. Vogel E, Köcher M, Schmickler H, Lex J. Porphycene—a Novel Porphin Isomer. Chem. Int. Ed. Engl. 1986; 25: 257-9.

2. Waluk J. ChemInform Abstract: Structure, Spectroscopy, Photophysics, and Tautomerism of Free-Base Porphycenes and Other Porphyrin Isomers, World Scientific Publishing Co. Pte. Ltd.: 2010; p 359-435.

Supporting information 3. Absorption and Photophysical Data of the TBPoS-OH (OH) and TBPoS-2OH (2OH).

Comp

Abs[a]

Em[b]

ΦF[c]

τF[d]

ΦΔ[e]

logPh

OH

650(3.87)

614 (2.76)

570 (2.92)

374 (9.60)

666

0.35

8.12

0.45[f]

0.56[g]

5.6

2OH

644 (3.58)

614 (3.18)

575 (2.75)

378 (8.47)

661

0.16

5.49

0.56f

0.68g

2.71

[a] Absorption abs/nm (ε×10-4/ M-1cm-1). [b] emission (λem / nm). [c] fluorescence quantum yield (λem=550-800 nm, λex =365 nm). [d] fluorescence lifetime (in ns) measured in CHCl3. [e] 1O2 quantum yield, measured in terms of its phosphorescence emission intensity at 1270 nm relative to using the following reference compounds, meso-tetraphenylporphyrin, H2TPP (f, ΦΔ = 0.55 ± 0.11, λex = 400 nm, CHCl3) and Zn(II) tetraphenylporphyrin, ZnTPP (g, ΦΔ = 0.73 ± 0.11, λex = 355 nm, toluene). [h] Obtained by using the ALOGPS 2.1 software (www.vcclab.org/lab/alogps/).

Supporting information 4. Comparison of the Oxidation and Reduction Potentials (volt vs. SCE) of Different Porphycene Derivative.

Compound

Oxidation

Reduction

ΔE=E3- E1

E1/2(3)

E1/2(4)

E1/2(1)

E1/2(2)

t-Bu4Po

+1.02

+1.43

–0.95

–1.39

1.97

TBPoS-OH

+1.18

+1.58a

–0.64

–0.97

1.82

TBPoS-2OH

+1.52a

-----

–0.43

–0.75

1.95

Pob

+1.0

-----

–0.73

–1.07

1.73

TPrPob

+0.97

+1.44

–0.90

–1.27

1.87

TPPob

+1.08

-----

–0.73

–0.98

1.81

TPPb

+0.95

+1.28

–1.05

–1.47

2.0

aPeak potential (vs. Ag|AgCl), bData from reference 28.

Supporting information 5. Comparison of the Dark Cytotoxicity (Dark LD50) and the Photo-Cytotoxicity (LD50) of TBPoS-OH (OH) and TBPoS-2OH (2OH) Towards Different Cell Lines under Hypoxic (H) and Normoxic (N) Conditions.

Dark LD50 (μM)

Light LD50 (nM)

PTIa (HeLa)

HeLa

KYSE 70

A375

B16

HeLa

KYSE 70

A375

B16

Dark

N

H

N

H

N

H

N

H

N

H

OHb

>100

>100

>100

>100

15

49

31

51

192

324

323

360

> 6666

> 2040

2OH

24

28

28

57

69

329

5

24

23

57

168

370

348

73

aPTI = Dark LD50 / Light LD50

bThe maximum solubility of TBPoS-OH was ca. 100 μM in 1‰ DMSO-PBS.

Supporting information 6

Hypoxic PDT Setup used in this study

Supporting information 6a. Photograph of our Hypoxic PDT setup.

Evaluation of our Hypoxic PDT Setup and Hypoxia Protocol

Supporting information 6b. Change of HIF-1α level in HeLa cells incubated for 3 and 4 h under normoxic and hypoxic conditions. C = Control (Normoxic); T = Treatment (Hypoxic).

Supporting information 7

Supporting information 7a. HPLC analysis of TBPoS-OH buffer containing dimethyl sulphoxide (1.0 mg/mL).

Supporting information 7b. HPLC analysis of TBPoS-OH in phosphate buffer saline.

Supporting information 7c. HPLC analysis of TBPoS-2OH buffer containing dimethyl sulphoxide (1.0 mg/mL).

Supporting information 7d. HPLC analysis of TBPoS-2OH in phosphate buffer saline.