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Supporting Information

Materials and general methods:

Chemicals: Fmoc-amino acids were obtained from GL Biochem (Shanghai, China). Phenothiazine was

obtained from A&K (Beijing, China). Naphthalene acetic acid was obtained from Aladdin (Shanghai,

China). 2-Cl-trityl chloride resin was obtained from Nankai University resin Co. Ltd. All the other

starting materials were obtained from Alfa. Commercially available reagents were used without further

purification, unless noted otherwise. All other chemicals were reagent grade or better.

General methods: The synthesized compounds were characterized by 1H NMR (Bruker ARX 300) using

DMSO-d6 as the solvent and ESI-MS spectrometric analyses were performed at the Thermo Finnigan

LCQ AD System. HPLC was conducted at LUMTECH HPLC (Germany) system using a C18 RP column

with MeOH (0.1% of TFA) and water (0.1% of TFA) as the eluents, LC-MS was conducted at the

LCMS-20AD (Shimadzu) system, and rheology was performed on an AR 2000ex (TA instrument) system

using a parallel plates (40 mm) at the gap of 500 μm. TEM was done on a Tecnai G2 F20 system,

operating at 200 kV. Confocal microscopy images were obtained on a Leica TCS SP5 system (Germany).

Fourier transform infrared (FT-IR) spectra were scanned on a bio-rad FTS6000 FT-IR spectrophotometer

(America). Circular dichroism (CD) data were collected on a Jasco J-715 CD spectrometer (Japan).

Syntheses and characterizations:

3-(Phenothiazin-10-yl)acetic acid synthesis: procedure was modified from the literature1. Phenothiazine

(1) (509 mg, 2.56 mmol), potassium hydroxide (171 mg, 2.61 mmol, 85%, freshly ground), and sodium

iodide (39 mg, 0.26 mmol) were combined under argon in 10 mL dry DMF. After 5 min stirring at rt, the

ethyl bromoacetate (7.7 mmol) was added dropwise. The reaction mixture was heated at 50 ºC for 48 h.

After the reaction mixture had cooled at rt for 30 min, it was diluted with DCM (15 mL) and washed with

water (3 × 10 mL). The combined organic layers were dried over magnesium sulfate, filtered, and rotary-

evaporated to give a yellow liquid (1.97 g). The crude product was purified on a flash silica column with

EtOAc/hexanes (4% to 8% to 15%) as eluent to give the pure compound of 2. The ester 2 was then

hydrolyzed in EtOH/MeOH/4M-KOH (25:25:8, 116 mL) at 60 oC for 30 min. After rotary-evaporating,

the reaction mixture was poured into water (100 mL) and acidified by slow addition of 10 % citric acid

until the salt was converted completely to the free acid. The resulting crystal was filtered and washed with

cold water. The crude acid was re-crystallized from ethanol to afford the final product of 3. 1H NMR

(300MHz, DMSO-d6) δ7.08-7.17 (m, 4H), 6.90-6.96 (t, 2H), 6.68-6.72 (d,J=8.12Hz, 2H), 4.55 (s, 2H).

Electronic Supplementary Material (ESI) for Chemical CommunicationsThis journal is © The Royal Society of Chemistry 2013

Scheme S-1. Synthetic pathway for the preparation of 3-(Phenothiazin-10-yl)propionic acid

ppm (f1)0.01.02.03.04.05.06.07.0

0

5000

7.1

62

7.1

44

7.1

23

7.1

10

7.0

94

6.9

49

6.9

31

6.9

12

6.7

09

6.6

89

4.5

49

2.0

0

2.0

1

2.1

0

4.2

0

Fig. S-1. 1H-NMR of 3-(Phenothiazin-10-yl)acetic acid

0.0

0.5

1.0

1.5

2.0

mV(x1,000)CH1 (220nm)

Fig. S-2. LC-MS of 3-(Phenothiazin-10-yl)acetic acid

258.70


Peptide systhesis: The peptide derivative was prepared by solid phase peptide synthesis (SPPS) using 2-

chlorotrityl chloride resin and the corresponding N-Fmoc protected amino acids with side chains properly

protected by a tert-butyl group. The first amino acid was loaded on the resin at the C-terminal with the

loading efficiency about 0.5 mmol/g. 20% piperidine in anhydrous N,N’-dimethylformamide (DMF) was

used during deprotection of Fmoc group. Then the next Fmoc-protected amino acid was coupled to the

free amino group using O-(Benzotriazol-1-yl)-N,N,N’,N’-tetramethyluroniumhexafluorophosphate

(HBTU) as the coupling reagent. The growth of the peptide chain was according to the established Fmoc

SPPS protocol. At the final step, Naphthalene acetic acid, 3-(Phenothiazin-10-yl)propionic acid or

benzyloxycarbony were used to attach on the peptide. After the last coupling step, excessive reagents

were removed by a single DMF wash for 5 minutes (5 mL per gram of resin), followed by five steps of

washing using DCM for 2 min (5 mL per gram of resin). The peptide derivative was cleaved using 95%

of trifluoroacetic acid with 2.5% of TMS and 2.5% of H2O for 30 minutes. After rotary-evaporate process,

20 mL per gram of resin of ice-cold diethylether was then added to cleavage reagent. Afterward the

supernatant was decanted and the resulting solid was dissolved in DMSO for HPLC separation.

Preparation of Fmoc-GFFY: 1H NMR (300MHz, DMSO-d6) δ 9.21 (s, 1H), 8.15-8.27 (m, 2H), 7.84-

7.95 (m, 3H), 7.64-7.71 (d, J=7.37Hz, 2H), 7.44-7.51 (t, 1H), 7.35-7.43 (t, 2H), 7.26-7.33 (t, 2H), 7.18-

7.24 (d, 4H), 7.09-7.17 (m, 6H), 6.98-7.04 (d, J=8.40, 2H), 6.60-6.68 (d, J=8.42, 2H), 4.44-4.59 (m, 2H),

4.32-4.41 (m, 1H), 4.15-4.27 (m, 3H), 3.42-3.65 (m, 2H), 2.63-3.05 (m, 6H). MS: calc. M+ = 754.3,

obsvd. (M+1)+ = 755.3. HR-MS: (M+1)

+ = 755.3070.


ppm (t1)0.01.02.03.04.05.06.07.08.09.0

0

100

200

300

400

500

9.2

11

8.2

41

8.2

15

8.1

72

7.9

29

7.9

02

7.8

84

7.8

59

7.6

90

7.6

66

7.4

77

7.4

56

7.4

17

7.3

93

7.3

70

7.3

20

7.2

95

7.2

71

7.2

25

7.2

10

7.1

34

7.0

24

6.9

96

6.6

55

6.6

27

4.5

52

4.5

08

4.4

79

4.3

75

4.3

56

4.2

43

4.2

25

4.1

95

3.6

27

3.6

07

3.5

71

3.5

51

3.5

03

3.4

83

3.4

27

3.0

24

2.9

52

2.9

07

2.8

55

2.8

29

2.7

19

2.6

41

2.0

0

2.2

3

0.9

3

1.8

5

2.8

7

1.7

8

0.8

42

.02

1.9

4

5.7

94

.03

1.9

30

.97

2.7

3

1.9

6

6.1

1

Fig. S-3. 1H-NMR of Fmoc-GFFY

Fig. S-4. HR-MS spectrum of Fmoc-GFFY


Preparation of Nap-GFFY:1H NMR (300MHz, DMSO-d6) δ 9.22 (s, 1H), 8.14-8.28 (m, 3H), 7.97-8.04

(d, J=8.41Hz, 1H), 7.77-7.88 (m, 3H), 7.73 (s, 1H), 7.36-7.50 (m, 3H), 7.26-7.33 (t, 2H), 7.18-7.23 (d,

4H), 7.10-7.17 (m, 6H), 6.98-7.05 (d, J=8.45, 2H), 6.61-6.68 (d, J=8.42, 2H), 4.43-4.60 (m, 2H), 4.31-

4.41 (m, 1H), 3.50-3.76 (m, 4H), 2.60-3.05 (m, 6H). MS: calc. M+ = 700.3, obsvd. (M+1)

+ = 701.3. HR-

MS: (M+1)+ = 701.2978.

ppm (f1)0.01.02.03.04.05.06.07.08.09.0

0

100

200

300

400

500

9.2

16

8.2

38

8.1

83

8.1

55

8.0

23

7.9

95

7.8

28

7.8

13

7.7

85

7.7

26

7.4

64

7.4

52

7.4

40

7.4

07

7.3

79

7.2

14

7.2

00

7.1

40

7.0

29

7.0

01

6.6

60

6.6

32

4.5

47

4.5

20

4.4

91

4.4

79

4.4

48

4.3

77

4.3

58

3.7

47

3.7

28

3.6

91

3.6

72

3.6

11

3.5

73

3.5

17

3.0

17

2.9

72

2.9

58

2.9

11

2.8

57

2.8

30

2.7

66

2.7

22

2.6

89

2.6

56

2.0

0

2.0

45

.96

3.9

3

2.9

7

0.9

82

.96

0.9

0

2.8

9

0.8

7

1.9

00

.96

3.9

6

6.1

6

Fig. S-5. 1H-NMR of Nap-GFFY


Fig. S-6. HR-MS spectrum of Nap-GFFFY

Preparation of PTZ-GFFY: 1H NMR (300MHz, DMSO-d6) δ 9.23 (s, 1H), 8.00-8.48 (m, 4H), 7.20-7.26

(d, J=4.19Hz, 4H), 7.11-7.19 (m, 6H), 6.99-7.09 (m, 6H), 6.83-6.92 (t, 2H), 6.60-6.73 (m, 4H), 4.48-4.63

(m, 2H), 4.30-4.46 (m, 3H), 3.73-3.87 (m, 1H), 3.57-3.70 (m, 1H), 2.90-3.07 (m, 3H), 2.62-2.89 (m, 3H).

MS: calc. M+ = 771.3, obsvd. (M+1)

+ =772.3. HR-MS: (M+1)

+ = 772.2792.


ppm (t1)0.01.02.03.04.05.06.07.08.09.0

0

100

200

300

400

500

600

700

8009

.22

8

8.3

72

8.2

28

8.0

99

8.0

72

7.2

27

7.2

13

7.1

64

7.1

36

7.0

57

7.0

32

7.0

03

6.9

01

6.8

76

6.8

51

6.6

97

6.6

69

6.6

57

6.6

30

4.5

68

4.5

54

4.5

39

4.5

26

4.5

11

4.4

30

4.3

71

4.3

53

3.8

33

3.8

13

3.7

76

3.7

58

3.6

72

3.6

55

3.6

16

3.5

99

3.0

37

3.0

23

2.9

81

2.9

36

2.8

62

2.8

35

2.7

95

2.7

65

2.7

22

2.6

91

2.6

46

4.0

0

0.8

8

6.1

6

1.9

6

3.9

2

1.9

2

0.8

8

0.9

9

3.1

8

3.2

8

2.8

1

4.1

86

.10

Fig. S-7. 1H-NMR of PTZ-GFFY

Fig. S-8. HR-MS spectrum of PTZ-GFFY


Preparation of Cbz-GFFY: 1H NMR (300MHz, DMSO-d6) δ 9.22 (s, 1H), 8.15-8.27 (m, 2H), 7.86-7.93

(d, J=8.26Hz, 1H), 7.35-7.41 (t, 1H), 7.28-7.35 (m, 4H), 7.19-7.25 (d, 4H), 7.09-7.18 (m, 6H), 6.98-7.04

(d, J=8.42, 2H), 6.61-6.67 (d, J=8.41, 2H), 4.99 (s, 2H), 4.43-4.60 (m, 2H), 4.32-4.41 (m, 1H), 3.47-3.64

(m, 2H), 2.63-3.05 (m, 6H). MS: calc. M+ = 666.3, obsvd. (M+1)

+ =667.3. HR-MS: (M+1)

+ = 667.2766.

ppm (t1)0.01.02.03.04.05.06.07.08.09.0

0

500

9.2

15

8.2

46

8.2

21

8.2

06

8.1

78

7.9

08

7.8

81

7.3

99

7.3

79

7.3

59

7.3

20

7.3

08

7.2

30

7.2

16

7.1

38

7.0

25

6.9

97

6.6

58

6.6

30

4.9

94

4.5

53

4.5

40

4.5

10

4.4

88

4.4

75

4.4

03

4.3

78

4.3

59

4.3

33

3.6

24

3.6

04

3.5

68

3.5

48

3.5

04

3.4

83

3.0

25

2.9

76

2.9

53

2.9

04

2.8

56

2.8

29

2.7

99

2.7

84

2.7

67

2.7

22

2.6

91

2.6

76

2.6

45

2.0

0

2.0

16

.07

4.1

44

.10

0.9

7

0.9

3

1.9

1

0.8

1

1.7

7

2.0

00

.99

6.2

4

2.3

0

Fig. S-9. 1H NMR of Cbz-GFFY


Fig. S-10. HR-MS of Cbz-GFFY

Formation of the gels: The gelators with one equiv. of Na2CO3 (to neutralize the terminal carboxylic

acid on the peptide) were suspended in PBS buffer (pH = 7.4). The suspensions were then heated in a hot

water bath to make the peptides totally dissolved. Hydrogels would form after cooling back to room

temperature within 10 minutes.

Fig. S-11. The rheological measurement with the mode of dynamic strain sweep for the gel of Fmoc-

GFFY


Fig. S-12. The rheological measurement with the mode of dynamic strain sweep for the gel of Nap-GFFY

Fig. S-13. The rheological measurement with the mode of dynamic strain sweep for the gel of PTZ-GFFY


Fig. S-14. Rheological measurements in the dynamic frequency sweep mode for the gels containing 0.2

wt% of gelators at a strain of 0.5%


Fig. S-15. FT-IR of freeze-dried hydrogels

1. S. Fanni, T. E. Keyes, S. Campagna and J. G. Vos, Inorg. Chem., 1998, 37, 5933-5935; P. Hanson, W. J. Isham, R. J.

Lewis and W. A. Stockburn, J. Chem. Soc., Perkin Trans. 2, 1981, 1492-1500;


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