Upconversion Nanotheranostic Agent Activated by Hypoxia

Combined with NIR Irradiation for Selective Hypoxia

Imaging and Tumour Therapy

Hongliang Li,† Weiyan Lei,† Jianong Wu, Shenghui Li, Guoqiang Zhou,* Dandan Liu, Xinjian Yang, Shuxiang Wang,* Zhenhua Li, and Jinchao Zhang*College of Chemistry & Environmental Science, Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of Ministry of Education, Chemical Biology Key Laboratory of Hebei Province, Hebei University, Baoding, 071002, China

zhougq1982@163.com, (tel)++86 312 507 9359, (fax) ++86 312 507 9359; wsx@hbu.edu.cn; (tel) ++86 312 507 9359, (fax) ++86 312 507 9359; jczhang6970@163.com, (tel) ++86 312 507 9525; (fax) ++86 312 507 9525

Supplementary Information

Electronic Supplementary Material (ESI) for Journal of Materials Chemistry B.This journal is © The Royal Society of Chemistry 2018

1. The characterization of FDU-CAE-NO2 and UCNP-CAE-FDU/NO2

2-(4-Nitrobenzyloxy)-4-(prop-2-ynyloxy)benzaldehyde (2) : A yellow solid, Mp:

CHO

O

ONO2

163.3-164.8 oC; 1H-NMR (600 MHz, DMSO-d6): δ 10.31 (s, 1 H), 8.28 (d, 2 H, J = 8.4 Hz, ), 7.80 (d, 2 H, J =

8.4 Hz), 7.73 (d, 1 H, J = 8.4 Hz), 6.86 (d, 1 H, J = 2.4 Hz), 6.76 (d, 1 H, J = 8.4 Hz), 5.46 (s, 2 H), 4.94 (d, 2 H, J

= 2.4 Hz), 3.64 (s, 1 H). 13C NMR (DMSO-d6, 150 MHz, ppm): δ 187.42, 163.66, 161.66, 147.10, 144.15,

130.18, 128.19, 123.68, 118.99, 107.70, 100.66, 78.90, 78.38, 68.67, 56.04. HRMS (ESI) m/z calcd for

C17H13NO5 [M+H]+ 312.08665, found 312.08673.

(E)-tert-Butyl-3-(2-(4-nitrobenzyloxy)-4-(prop-2-ynyloxy)phenyl) acrylate (4) : A

OO

NO2

O

O

white solid. Mp: 145.7-148.3 oC; 1H-NMR (DMSO-d6, 600 MHz, ppm): δ 8.29 (d, 2H, J = 9.0 Hz), 7.83 (d, 1 H,

J = 15.6 Hz), 7.74 (d, 2 H, J = 9.0 Hz), 7.72 (d, 1H, J = 9.0 Hz), 6.76 (d, 1H, J = 2.4 Hz), 6.67 (dd, 1H, J = 9.0,

2.4 Hz,) 6.42 (d, 1H, J = 15.6 Hz) , 5.39 (s, 2H), 4.86 (d, 2H, J = 2.4 Hz), 3.60 (s, 1H) , 1.47 (s, 9H). 13C NMR

(DMSO-d6, 150 MHz, ppm): δ 166.00, 160.20, 157.53, 147.11, 144.40, 137.70, 129.67, 128.22, 123.67,

117.71, 116.37, 107.27, 100.73, 79.46, 78.74, 78.51, 68.64, 55.73, 27.81. HRMS (ESI) m/z calcd for

C23H23NO6 [M+H]+ 410.15981, found 410.16003.

(E)-3-(2-(4-Nitrobenzyloxy)-4-(prop-2-ynyloxy)phenyl)acrylic acid (5) : A white solid,

OO

OH

O

NO2

Mp: 197.6-199.5 oC. 1H-NMR (DMSO-d6, 600 MHz, ppm): δ 8.28 (d, 2 H, J = 8.4 Hz), 7.73 (d, 2 H, J = 8.4 Hz),

7.50 (d, 1 H, J = 8.4 Hz), 7.47 (d, 1 H, J = 15.6 Hz), 6.69 (d, 1 H, J = 2.4 Hz), 6.60 (dd, 1 H, J = 9.0, 2.4 Hz),

6.30 (d, 1 H, J = 15.6 Hz) , 5.34 (s, 2 H), 4.79 (d, 2 H, J = 2.4 Hz), 3.56 (s, 1 H). 13C NMR (DMSO-d6, 150 MHz,

ppm): δ 167.99, 160.14, 157.54, 147.18, 144.35, 138.22, 128.39, 123.73, 117.22, 116.48, 107.29, 100.76,

78.76, 78.51, 68.64, 55.74. HRMS (ESI) m/z calcd for C19H15NO6 [M+H]+ 354.09721, found 354.09723.

FDU-CAE-NO2 : A white solid. Mp: 143.4-146.3 oC. 1H NMR (DMSO-d6, 600

OO

O

O

NO2

NHN

OO

HO

O

F

MHz, ppm): δ 11.88 (s, 1 H), 8.28 (d, 2 H, J = 8.7 Hz), 7.92 (d, 1 H, J = 16.0 Hz), 7.87 (d, 1 H, J = 6.8 Hz), 7.75

(d, 1 H, J = 8.8 Hz), 7.72 (d, 2 H, J = 8.7 Hz), 6.77 (d, 1 H, J = 2.0 Hz), 6.69 (dd, 1 H, J = 8.8, 2.0 Hz), 6.60 (d, 1

H, J = 16.0 Hz), 6.14 (t, 1 H, J = 6.4 Hz), 5.51 (d, 1 H, J = 4.4 Hz), 5.40 (d, 2 H, J = 3.2 Hz), 4.87 (d, 2 H, J = 2.0

Hz), 4.40-4.28 (m, 3 H), 4.01 (s, 1 H), 3.61 (s, 1 H), 2.24-2.09 (m, 2 H). 13C NMR (DMSO-d6, 150 MHz, ppm):

δ 166.47, 160.54, 157.03, 157.03, 156.77, 148.86, 147.09, 144.29, 141.12, 139.43, 138.83, 130.21, 128.26,

124.53, 124.19, 123.71, 116.12, 115.53, 107.34, 100.73, 84.55, 83.97, 78.71, 78.59, 70.00, 68.65, 63.67,

55.77. HRMS (ESI) m/z calcd for C28H24N3FO10 [M+H]+ 582.15185, found 582.15216.

7-Propargyloxycoumarin (CM) : A white solid yield, Mp: 70.3-71.8 oC. 1H NMR (CDCl3, O OO

600 MHz, ppm): δ 7.64 (d, 1 H, J = 9.6 Hz), 7.41 (d, 1 H, J = 8.4 Hz), 6.94 (s, 1 H), 6.92 (d, 1 H, J = 8.4 Hz),

6.29 (d, 1 H, J = 9.6 Hz), 4.77 (d, 2 H, J = 2.4 Hz), 2.56 (s, 1 H). 13C NMR (CDCl3, 150 MHz, ppm): δ 160.98,

160.55, 155.66, 143.27, 128.84, 113.67, 113.20, 113.05, 102.16, 77.38, 76.57, 56.22. HRMS (ESI) m/z calcd

for C12H8O3 [M+H]+ 201.05462, found 201.05461.

2. The size of UCNP-CAE-FDU/NO2 and UCNP analyzed by DLS and TEM

The samples for TEM and DLS were prepared by dispersing UCNP-CAE-FDU/NO2 (200 μg/mL) in PBS

(25 mmol/L, pH 7.4) with intermittent ultrasonic by a needle type ultrasonic instrument. The samples of

UCNP were dispersed in cyclohexane (20 μg/mL). The TEM sample was prepared by dropping on the

surface of a copper grid and negative staining for 30 s by a droplet of phosphotungstic acid. The DLS of the

samples were measured using a Nano-ZS system in disposable cuvettes.

3. HPLC and HRMS analysis

Figure S1. HPLC profiles of a) FDU; b) FDU-CAE-NO2; c) CM; d) solution of FDU-CAE-NO2 incubated with

Na2S2O4 and then illuminated by UV light at 365 nm; e) UCNP-CAE-FDU/NO2 incubated with Na2S2O4 and

then illuminated by NIR light at 980 nm

Figure S2. HRMS of the solution of FDU-CAE-NO2 incubated with Na2S2O4 and then illuminated by UV-light at 365 nm

[M+H]+(C12H9O3): 201.05462

O OO

[M+H]+(C9H12FN2O5): 247.07248

NH

N

OO

HO

HO O

F

OOO

O

NO2

NHN

OO

HO

O

F

[M+H]+(C28H25FN3O10): 582.15185

Figure S3. HRMS of the mixture of UCNP-CAE-FDU/NO2 with Na2S2O4 and then illuminated by NIR-light at 980 nm

4. NMR, IR and HRMS spectra

CHO

OO

NO2

0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.5f1 (ppm)

0.88

2.08

2.21

1.05

1.00

1.09

2.11

2.22

1.00

3.64

4.93

4.94

5.45

6.75

6.76

6.86

6.86

7.72

7.74

7.81

8.27

8.28

10.3

1

Figure S4. 1H NMR of 2-(4-nitrobenzyloxy)-4-(prop-2-ynyloxy)benzaldehyde (2)

[M+H]+(C9H12FN2O5): 247.07248

NH

N

OO

HO

HO O

F

0102030405060708090100110120130140150160170180190f1 (ppm)

56.0

41

68.6

65

78.3

7878

.904

100.

655

107.

697

118.

991

123.

679

128.

186

130.

183

144.

148

147.

100

161.

655

163.

664

187.

420

CHO

OO

NO2

Figure S5. 13C NMR of 2-(4-nitrobenzyloxy)-4-(prop-2-ynyloxy)benzaldehyde (2)

CHO

OO

NO2

Figure S6. HRMS of 2-(4-nitrobenzyloxy)-4-(prop-2-ynyloxy)benzaldehyde (2)

Figure S7. IR of 4-nitrobenzyloxy-4-diethylaminobenzaldehyde (2)

0.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f1 (ppm)

9.41

0.92

2.08

2.02

1.00

1.22

1.01

1.13

2.02

1.01

2.13

1.47

1

3.59

5

4.85

74.

860

5.39

3

6.40

76.

434

6.66

16.

665

6.67

6

6.75

87.

708

7.72

27.

730

7.74

57.

814

8.28

18.

296

OO

NO2

OO

Figure S8. 1H NMR of (E)-tert-butyl-3-(2-(4-nitrobenzyloxy)-4-(prop-2-ynyloxy)phenyl)acrylate (4)

0102030405060708090100110120130140150160170f1 (ppm)

27.8

10

55.7

25

68.6

43

78.5

0578

.741

79.4

56

100.

734

107.

271

116.

369

117.

714

123.

658

128.

215

129.

661

137.

695

144.

397

147.

114

157.

525

160.

195

166.

001

OO

NO2

OO

Figure S9. 13C NMR of (E)-tert-butyl-3-(2-(4-nitrobenzyloxy)-4-(prop-2-ynyloxy)phenyl)acrylate (4)

OO

NO2

OO

Figure S10. HRMS of (E)-tert-butyl-3-(2-(4-nitrobenzyloxy)-4-(prop-2-ynyloxy)phenyl)acrylate (4)

Figure S11. IR of (E)-tert-butyl-3-(2-(4-nitrobenzyloxy)-4-(prop-2-ynyloxy)phenyl)acrylate (4)

OOOH

O

NO2

2.02.53.03.54.04.55.05.56.06.57.07.58.08.5f1 (ppm)

1.04

2.00

2.00

1.00

1.00

0.98

0.94

1.03

2.01

2.01

3.55

6

4.78

74.

791

5.34

2

6.29

16.

317

6.59

26.

595

6.60

6

6.68

47.

453

7.47

97.

493

7.72

27.

736

8.26

78.

281

Figure S12. 1H NMR of (E)-3-(2-(4-nitrobenzyloxy)-4-(prop-2-ynyloxy)phenyl)acrylic acid (5)

0102030405060708090100110120130140150160170180f1 (ppm)

55.7

36

68.6

38

78.5

1378

.756

100.

758

107.

293

116.

481

117.

222

123.

729

128.

390

138.

222

144.

351

147.

177

157.

536

160.

143

167.

990

OOOH

O

NO2

Figure S13. 13C NMR of (E)-3-(2-(4-nitrobenzyloxy)-4-(prop-2-ynyloxy)phenyl)acrylic acid (5)

OO

NO2

OHO

Figure S14. HRMS of (E)-3-(2-(4-nitrobenzyloxy)-4-(prop-2-ynyloxy)phenyl)acrylic acid (5)

0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.511.5f1 (ppm)

2.06

1.03

1.00

3.09

2.00

2.01

1.00

1.04

1.07

0.97

1.01

3.00

1.01

1.03

2.07

1.01

2.09

42.

244

3.60

7

4.01

04.

282

4.87

0

5.50

65.

517

6.12

9

6.57

76.

617

6.77

07.

707

8.26

68.

287

11.8

65

OOO

O

NO2

NHNO

O

HO

O

F

Figure S15. 1H NMR of FDU-CAE-NO2

OOO

O

NO2

NHN

OO

HO

O

F

0102030405060708090100110120130140150160f1 (ppm)

55.7

68

63.6

73

68.6

5470

.004

78.5

90

83.9

7184

.545

100.

730

107.

341

115.

332

116.

118

123.

707

128.

257

130.

208

138.

825

139.

436

141.

123

144.

294

156.

774

157.

034

160.

544

166.

465

Figure S16. 13C NMR of FDU-CAE-NO2

OOO

O

NO2

NHNO

O

HOOF

Figure S17. HRMS of FDU-CAE-NO2

Figure S18. IR of FDU-CAE-NO2

0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0f1 (ppm)

1.00

2.23

1.07

1.03

1.01

1.08

1.08

2.58

5

4.76

64.

770

6.27

86.

294

6.91

26.

926

6.94

3

7.40

1

7.64

77.

663

O OO

Figure S19. 1H NMR of CM

0102030405060708090100110120130140150160f1 (ppm)

56.2

20

76.5

6777

.379

102.

159

113.

049

113.

201

113.

671

128.

841

143.

268

155.

655

160.

552

160.

981

O OO

Figure S20. 13C NMR of CM

O OO

Figure S21. HRMS of CM

Figure S22. IR of CM

Figure S23. IR of UCNPs

Figure S24. IR of UCNP-CAE-FDU/NO2