Kohei Iijima, Paula Lusarreta Parga, Utsa Karmakar, Ferran Nadal-Bufi, Alejandro J. Brenes, Iris Ponton, Alex von Kriegsheim, Kazuya Kikuchi, Marc Vendrell. “Activity-guided identification of pro-inflammatory neutrophils with dual enzyme- and metabolite-activatable fluorescent probes.” ChemRxiv (). https://doi.org/10.26434/chemrxiv.15007821/v1

4-Iodoresorcinol
¹H NMR (500 MHz, CDCl₃) δ 7.45 (d, J = 8.6 Hz, 1H), 6.54 (d, J = 2.7 Hz, 1H), 6.27 (dd, J = 8.6, 2.7 Hz, 1H), 5.30 (bs, 2H)
¹³C NMR (125 MHz, CDCl₃) δ 157.7 155.8, 138.5, 110.6, 102.8, 74.7
HRMS (ESI) m/z calcd. for C₆H₄IO₂ [M − H]⁻: 234.9262; found: 234.9269
NMR spectra Image data were not transcribed
SI, pp. 3, 43–44
7-(dimethylamino)-2-iodo-3H-phenoxazin-3-one
Identifier 1
¹H NMR (500 MHz, MeOD-d₄) δ 7.77 (d, J = 9.5 Hz, 1H), 7.69 (d, J = 9.2 Hz, 1H), 7.34 (dd, J = 9.6, 2.6 Hz, 1H), 7.03 (d, J = 2.7 Hz, 1H), 6.98 (d, J = 2.7 Hz, 1H), 3.41 (s, 6H)
¹³C NMR (125 MHz, CDCl₃) δ 179.5, 154.3, 150.7, 146.9, 142.9, 140.8, 132.2, 126.9, 111.5, 111.5, 103.4, 96.7, 40.7
HRMS (ESI) m/z calcd. for C₁₄H₁₁IN₂O₂Na [M + Na]⁺: 388.9757; found: 388.9773
NMR spectra Image data were not transcribed
SI, pp. 4, 45–46
Fmoc-Trp-oxazone
Identifier 2
¹H NMR (500 MHz, acetone-d₆) δ 10.87 (s, 1H), 7.82 – 7.76 (m, 4H), 7.57 (dd, J = 7.4, 4.7 Hz, 2H), 7.49 (t, J = 8.8 Hz, 2H), 7.36 – 7.29 (m, 3H), 7.25 – 7.15 (m, 3H), 7.08 (t, J = 7.5 Hz, 1H), 6.88 (dd, J = 9.2, 2.5 Hz, 1H), 6.61 (d, J = 2.5 Hz, 1H), 6.33 (s, 1H), 4.76 (m, 1H), 4.15 – 4.05 (m, 3H), 3.64 (dd, J = 14.8, 5.0 Hz, 1H), 3.49 (dd, J = 14.7, 9.2 Hz, 1H), 3.22 (s, 6H)
¹³C NMR (125 MHz, acetone-d₆) δ 184.5, 173.9, 157.0, 155.4, 150.8, 147.5, 145.2, 145.0, 142.0, 140.9, 137.6, 137.4, 133.6, 132.6, 129.6, 128.7, 128.5, 128.0, 126.4, 126.3, 123.5, 120.7, 120.2, 120, 112.8, 112.4, 106.6, 97.3, 67.3, 55.5, 47.9, 41.3 40.8
HRMS (ESI) m/z calcd. for C₄₀H₃₃N₄O₆ [M + H]⁺: 665.2395; found: 665.2370
NMR spectra Image data were not transcribed
Absorption spectrum (5 µM, PBS, pH 7.4) Image data were not transcribed
SI, pp. 5, 23, 47–48
tert-butyl 4-iodo-3-methylbenzoate
Identifier 3
¹H NMR (500 MHz, CDCl₃) δ 7.86 (d, J = 8.2 Hz, 1H), 7.81 (d, J = 1.9 Hz, 1H), 7.45 (dd, J = 8.2, 1.9 Hz, 1H), 2.47 (s, 3H), 1.58 (s, 9H)
¹³C NMR (125 MHz, CDCl₃) δ 165.6, 152.3, 141.7, 139.1, 132.2, 130.4, 127.9, 106.9, 81.2, 28.3
HRMS (EI) m/z calcd. for C₁₂H₁₅IO₂ [M]⁺: 318.0111; found: 318.0117
NMR spectra Image data were not transcribed
SI, pp. 6, 49–50
tert-butyl 3-((tert-butylthio)methyl)-4-iodobenzoate
Identifier 5
¹H NMR (500 MHz, CDCl₃) δ 8.00 (d, J = 2.1 Hz, 1H), 7.86 (d, J = 8.5 Hz, 1H), 7.58 (dd, J = 8.5, 2.1 Hz, 1H), 3.90 (s, 2H), 1.58 (s, 9H), 1.43 (s, 9H)
¹³C NMR (125 MHz, CDCl₃) δ 165.2, 141.4, 139.9, 132.6, 131.0, 129.3, 106.1, 81.6, 43.6, 38.7, 31.0, 28.3
HRMS (ESI) m/z calcd. for C₁₆H₂₄O₂IS [M + H]⁺: 407.0536; found, 407.0529
NMR spectra Image data were not transcribed
SI, pp. 7, 51–52
9-oxo-9H-xanthene-3,6-diyl bis(trifluoromethanesulfonate)
Identifier 6
¹H NMR (500 MHz, CDCl₃) δ 8.45 (d, J = 8.8 Hz, 2H), 7.49 (d, J = 2.3 Hz, 2H), 7.35 (dd, J = 8.8, 2.3 Hz, 2H)
¹³C NMR (125 MHz, CDCl₃) δ 174.4, 156.5, 153.3, 129.5, 121.3, 118.6 (q, JC-F = 320 Hz), 118.1, 111.3
HRMS (ESI) m/z calcd. for C₁₅H₆F₆O₈S₂Cl⁻ [M + Cl]⁻: 526.9102; found: 526.9118
NMR spectra Image data were not transcribed
SI, pp. 8, 53–54
3,6-bis((diphenylmethylene)amino)-9H-xanthen-9-one
Identifier 7
¹H NMR (500 MHz, CDCl₃) δ 8.05 (d, J = 8.4 Hz, 2H), 7.77 (bs, 4H), 7.51 (bs, 2H), 7.44 (bs, 4H), 7.29 (bs, 6H), 7.16 (bs, 4H), 6.75 (d, J = 1.7 Hz, 2H), 6.66 (dd, J = 8.4, 1.7 Hz, 2H)
¹³C NMR (125 MHz, CDCl₃) δ 176.1, 169.3, 157.6, 157.0, 138.9, 135.6, 132.5, 131.5, 130.2, 129.7, 129.4, 128.5, 128.4, 128.3, 127.3, 117.7, 117.6, 108.4
HRMS (ESI) m/z calcd. for C₃₉H₂₇N₂O₂ [M + H]⁺: 555.2067; found: 555.2078
NMR spectra Image data were not transcribed
SI, pp. 9, 55–56
6-amino-9-(4-carboxy-2-methylphenyl)-3H-xanthen-3-iminium
Identifier Rh-Green
¹H NMR (500 MHz, MeOD-d₄) δ 8.15 (s, 1H), 8.09 (d, J = 7.9 Hz, 1H), 7.38 (d, J = 7.9 Hz, 1H), 7.07 (d, J = 9.0 Hz, 2H), 6.86 (d, J = 2.1 Hz 1H), 6.84 (m, 3H) 2.12 (s, 3H)
¹³C NMR (125 MHz, MeOD-d₄) δ 169.0, 161.7, 159.9, 158.9, 137.9, 137.8, 133.7, 132.9, 130.4, 128.4, 118.4, 114.4, 98.6, 19
HRMS (ESI) m/z calcd. for C₂₁H₁₇N₂O₃⁺ [M]⁺: 345.1234; found: 345.1229
NMR spectra Image data were not transcribed
Photophysical properties in H₂O Image data were not transcribed
SI, pp. 10, 26, 59–60
N-(9-(2-((2-carboxyethyl)(methyl)carbamoyl)phenyl)-6-(diethylamino)-3H-xanthen-3-ylidene)-N-ethylethanaminium
Identifier Rh-Red
¹H NMR (500 MHz, MeOD-d₄) δ 7.77 (m, 2H), 7.67 (m, 1H), 7.53 (m, 1H), 7.30 (d, J = 9.5 Hz, 2H), 7.07 (dd, J = 9.6, 2.5 Hz, 2H), 6.96 (d, J = 2.5 Hz, 2H), 3.69 (q, J = 7.2 Hz, 8H), 3.40 (t, J = 6.9 Hz, 2H), 2.91 (s, 3H), 2.02 (t, J = 6.9 Hz, 2H), 1.33 (t, J = 7.2 Hz, 12H)
¹³C NMR (125 MHz, MeOD-d₄) δ 174.6, 170.7, 159.3, 157.2, 156.9, 137.6, 133.3, 131.8, 131.7, 131.4, 131.0, 128.9, 115.3, 114.7, 97.5, 46.9, 44.8, 39.1, 31.9, 12.8
HRMS (ESI) m/z calcd. for C₃₂H₃₄N₃O₄⁺ [M]⁺: 528.2857; found: 528.2860
NMR spectra Image data were not transcribed
Photophysical properties in H₂O Image data were not transcribed
SI, pp. 11, 26, 61–62
4,4'-methylenebis(3-bromo-N,N-dimethylaniline)
Identifier 8
¹H NMR (500 MHz, CDCl₃) δ 6.97 (d, J = 2.7 Hz, 2H), 6.88 (d, J = 8.6 Hz, 2H), 6.61 (dd, J = 8.6, 2.7 Hz, 2H), 4.03 (s, 2H), 2.94 (s, 12H)
¹³C NMR (125 MHz, CDCl₃) δ 150.2, 131.1, 127.3, 125.8, 116.7, 112.0, 40.7, 39.8
HRMS (ESI) m/z calcd. for C₁₇H₂₁N₂Br₂ [M + H]⁺: 411.0066; found: 411.0065
NMR spectra Image data were not transcribed
SI, pp. 12, 57–58
3,7-bis(dimethylamino)-5,5-dimethyldibenzo[b,e]silin-10(5H)-one
Identifier Si-xanthone
¹H NMR (500 MHz, CDCl₃) δ 8.40 (d, J = 8.9 Hz, 2H), 6.84 (dd, J = 8.9, 2.6 Hz, 2H), 6.80 (d, J = 2.6 Hz, 2H), 3.09 (s, 12H), 0.48 (s, 6H)
¹³C NMR (125 MHz, CDCl₃) δ 185.4, 151.6, 140.6, 131.7, 129.8 114.4, 113.3, 40.2, −0.9
HRMS (ESI) m/z calcd. for C₁₉H₂₄N₂OSiNa [M + Na]⁺: 347.1550; found, 347.1537
NMR spectra Image data were not transcribed
SI, pp. 13, 63–64
N-(10-(4-carboxy-2-methylphenyl)-7-(dimethylamino)-5,5-dimethyldibenzo[b,e]silin-3(5H)-ylidene)-N-methylmethanaminium
Identifier Rh-NIR
¹H NMR (500 MHz, MeOD-d₄) δ 8.04 (d, J = 1.1 Hz, 1H), 8.01 (dd, J = 7.9, 1.1 Hz 1H), 7.37 (d, J = 2.9 Hz, 2H), 7.22 (d, J = 7.9 Hz, 1H), 7.04 (d, J = 9.7 Hz, 2H), 6.77 (dd, J = 9.7, 2.9 Hz, 2H), 3.35 (s, 12H), 2.09 (s, 3H), 0.62 (s, 3H), 0.60 (s, 3H)
¹³C NMR (125 MHz, MeOD-d₄) δ 170.4, 169.6, 155.8, 149.5, 144.2, 142.0, 137.4, 134.2, 132.4, 130.4, 128.1, 128.0, 122.3, 115.4, 40.9, 19.4, −1.1, −1.3
HRMS (ESI) m/z calcd. for C₂₇H₃₁N₂O₂Si⁺ [M]⁺: 443.2149; found: 443.2151
NMR spectra Image data were not transcribed
Photophysical properties in H₂O Image data were not transcribed
SI, pp. 14–15, 26, 65–66
3',7'-bis(dimethylamino)-5',5'-dimethyl-3H,5'H-spiro[benzo[c]thiophene-1,10'-dibenzo[b,e]siline]-5-carboxylic acid
Identifier cSiOCl
¹H NMR (500 MHz, MeOD-d₄) δ 8.10 (d, J = 1.2 Hz, 1H), 7.91 (dd, J = 8.1, 1.2 Hz, 1H), 6.98 (d, J = 8.1 Hz, 1H), 6.96 (t, J = 1.7 Hz, 2H), 6.62 (d, J = 1.7 Hz, 4H), 4.34 (s, 2H), 2.91 (s, 12H), 0.60 (s, 3H), 0.44 (s, 3H)
¹³C NMR (125 MHz, MeOD-d₄) δ 169.7, 156.1, 149.7, 144.7, 142.0, 136.0, 131.2, 130.8, 129.7, 128.7, 127.2, 117.5, 115.7, 71.0, 40.8, 37.9, 1.3, −2.5
HRMS (ESI) m/z calcd. for C₂₇H₃₁N₂O₂SiS [M + H]⁺: 475.1870; found: 475.1855
NMR spectra Image data were not transcribed
Fluorescence spectra (2 µM, PBS, pH 7.4; HOCl 0.1–3.0 µM; excitation 630 nm) Image data were not transcribed
SI, pp. 16–17, 34, 67–68
CatG-Green
Photophysical properties in H₂O Image data were not transcribed
HPLC-HRMS Calc. m/z: 1845.7220; image data were not transcribed
SI, pp. 26, 69
CatG-Red
Photophysical properties in H₂O Image data were not transcribed
HPLC-HRMS Calc. m/z: 2028.8843; image data were not transcribed
SI, pp. 26, 70
CatG-NIR
Photophysical properties in H₂O Image data were not transcribed
HPLC-HRMS Calc. m/z: 1943.8136; image data were not transcribed
SI, pp. 26, 71
CatG-cSiOCl
HPLC-HRMS Calc. m/z: 1975.7856; image data were not transcribed