Supporting Information. Amide -Cyclodextrin by Photoirradiation

Transcription

1 Supporting Information Switching between Supramolecular Dimer and Non- Threaded Supramolecular Self-Assembly of Stilbene Amide -Cyclodextrin by Photoirradiation Kazuhiro Yamauchi, Yoshinori Takashima, Akihito Hashidzume, Hiroyasu Yamaguchi and Akira Harada* Department of Macromolecular Science, Graduate School of Science, Osaka University, Toyonaka, Osaka , Japan. * Corresponding author. Telephone ; Fax

2 Page content 4 Materials, Measurements, Determination of Diffusion Coefficient 6 Preparation of mono-3-trans-stilbene amide--cd (3-trans-Sti--CD) and mono-3-cisstilbene amide--cd (3-cis-Sti--CD). (a) Preparation of 4-stilbene carboxylic acid. 7 (b) Preparation of 3-trans-Sti--CD. 8 (c) Preparation of 3-cis-Sti--CD. 9 Figure. Chromatogram of the mixture of 3-trans-Sti--CD and 3-cis-Sti--CD. 10 Figure S2. The 1 H-NMR spectrum of 3-trans-Sti--CD in DMSO-d 6 at 30 o C. Figure S3. The 1 H-NMR spectrum of 3-cis-Sti--CD in DMSO-d 6 at 30 o C. 11 Figure S4. The 1 H-NMR spectra of 3-cis-Sti--CDs in D 2 O at 30 o C a) after and b) before irradiation at 254nm 12 Figure S5. The circular dichroism spectra of 3-trans-Sti--CD in an aqueous solution at 30 o C. (1.0 mm) 13 Figure S6. The circular dichroism spectra of 3-cis-Sti--CD in an aqueous solution (black line), 3-cis-Sti--CD in MeOH (red line), and 3-cis-Sti--CD with -CD in an aqueous solution (blue line) at 30 o C. 13 Figure S7. The 1 H-NMR spectra of various concentrations of 3-cis-Sti--CDs in D 2 O at 30 o C. Figure S8. Concentration dependence of of 3-cis-Sti--CD at 30 o C. 14 Fgure S9. The 1 H-NMR spectra of 3-cis-Sti--CDs in D 2 O at 30 o C.(5.0 mm) a) before and b) after adding -CD(50 mm) 15 Figure 0. Concentration dependence of absorption maximum around 290 nm of 3-cis-Sti- -CD in aqueous solutions. S2

3 16 Figure 1. Stejskal-Tanner plots ln(i/i 0 ) vs. g 2 of 3-CiO--CD with increased in the concentration of 3-cis-Sti--CD (1 mm (a), 10 mm (b)25 mm, (c)50 mm (d), and 100 mm (e))at 30 o C in D 2 O. 19 Figure 2. Concentration dependence of diffusion coefficient values (D) of supramolecular complexes formed 2-CiO--CD (rhombus) and 3-cis-Sti--CD (circle) in D 2 O at 30 C by NMR 20 Figure 3. Turbo-ion spray mass spectrum of 3-cis-Sti--CD in an aqueous solution. 21 Table. Crystal data and collection parameters for 3-cis-Sti--CD.. S3

4 Materials -Cyclodextrin (-CD) was obtained from Junsei Chemical Co., Ltd. Triphenylphosphine was obtained from Wako Pure Chemical Industries, Ltd. 4-Iodobenzoic acid and paradium( ) acetate was obtained from Tokyo Chemical Industry Co., Ltd. Styrene, triethylamine, 1-hydroxybenzotriazole acid, and N, N -dicyclohexylcarbodiimide were obtained from Nacalai Tesque Inc. Measurements The 1 H-NMR spectra were recorded at 500 MHz with a JEOL JNM LA-500 NMR spectrometer. Chemical shifts were referenced to the solvent values ( 2.49 ppm for DMSO and 4.65 ppm for HOD). 2D NMR (COSY, TOCSY, ROESY) experiments were performed with D 2 O at 30 o C at 500 MHz with a JEOL JNM LA-500 NMR spectrometer. The preparative reversed phase chromatography was carried out with a Waters Delta 600 system (column: SunFireTM Prep C mm). Positive-ion matrix assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry experiments were performed using a Shimadzu / KRATOS Axima CFR Ver mass spectrometer calibrated by - cyano-4-hydroxycinnamic acid and insulin. Turbo ionization spray mass spectrometry measurements were performed by using a Applied Biosystems Q-Star XL mass spectrometer. S4

5 Determination of Diffusion Coefficient: The pulse field gradient spin-echo (PFGSE) NMR spectra were recorded at 600 MHz in D 2 O on a VARIAN INOVA NMR spectrometer at 30.0 C. The bipolar pulse pair stimulated echo (BPPSTE) sequence was applied for PFGSE NMR measurement. The pulsed gradients strength was increased from to 43.1 (gauss/cm). The time separation of pulsed field gradients and their duration were 0.10 and 1.0 K10-3 (s). The sample was not spin and the airflow was disconnected. The shape of the gradient pulse was rectangular, and its strength varied automatically during the course of the experiments. The D values were determined from the slope of the regression line ln (I / I o ) versus G 2, according to the Stejskal and Tanner s Equation (1). ln ( I / I 0 ) = - 2 G 2 2 ( - / 3- / 2 ) D (1) I / I 0 : observed spin echo intensity / intensity without gradients, G : gradient strength, : delay between the midpoints of the gradients, D : diffusion coefficient, : gradient length, : 90 o -180 o pluse distance. The calibration of the gradients was carried out by a diffusion measurement of H 2 O (D H2O = m 2 s -1 ) at 25 o C. According to the Stejskal and Tanner s reports, when ln I / I 0 versus g 2 were plotted, where I and g are the echo intensity and (pulsed) gradient strength, respectively, the slope of the line given by D/( / 3) 2 2. References (1) Stibs, P. Prog. NMR Spectrosc. 1987, 19, (2) Stejskal, E. O.; Tanner, J. E. J. Chem. Phys. 1965, 42, S5

6 Preparation of mono-3-trans-stilbene amide- -CD (3-trans-Sti- -CD) and 3-cis-stilbene amide-cd (3-cis-Sti- -CD) (a) Preparation of 4-stillbene carboxylic acid Styrene (1.68 g, 16.1 mmol) and 4-iodobenzoic acid (4.00 g, 11.3 mmol) were dissolved in N,Ndimethylformamide (DMF; 20 ml) and triethylamine (20 ml). The solution was refluxed in the presence of triphenylphosphine ( g, mmol) and palladium (II) acetate ( g, mmol) for 24 hours. After the removal of the solvent, ethyl acetate (200 ml) was added, and the soluble part was washed with three portions of water (60 ml). The organic layer was reprecipitated in hexane (400mL) to give 4-stilbene carboxylic acid. (pale brown powder, 1.58 g, 43.8%). 1 H NMR (500 MHz, DMSO-d6) H 12.8 (br s, 1H, OH), (d, J = 7.8 ዊዊ,2H, 3-position of stilbene), (d, J = 7.8ዊዊዊ 2H, 2-position of stilbene), (d, J = 7.8Hz, 2H, 2 position of stilbene), (t, J = 8.1Hz, 2H, 3 -position of stilbene), (d, J = 16Hz, 2H, olefin of stilbene), (m, J = 7.3 Hz, 1H, 4 -position of stilbene).. TLC (ethyl acetate/hexane 1:1): Rf = 0.30 (relative to the solvent front). IR (KBr), (O-H), 1680 (C=O), 1425 (O-H), 1290 (C-O) cm-1. FAB-MS, m/z, 224 (M+), 207, 178, 154, 135, 107. Mp oc. Anal. calced for C15H12O2ዊ0.17H2O: C,79.26; H,5.47. Found: C,79.27; H,5.49. S6

7 (b) Preparation of 3-trans-Sti- -CD. 4-Stilbene carboxylic acid (112 mg, mmol), dicyclohexyl carbodiimide (DCC; 320 mg, 1.55 mmol) and 1-hydroxybenzotriazole (HOBt; 97.0 mg, mmol) were added to dried DMF (50 ml) at 0 oc. After an hour, a solution of 3-NH2- -CD (490 mg, mmol) in dried DMF (50 ml) was added to the reaction mixture, and the mixture was stirred at 0 oc for two hours. It was allowed to warm to room temperature and stirred for 60 hours. The mixture was poured into acetone (500 ml). The precipitate was collected, and dissolved in water. The solution was applied to a DIAION HP-20 column. The column was flushed with water, then eluted with a water/methanol = 20/80 (v/v). The fraction was concentrated under reduced pressure to give pure 3-trans-sti- -CD (white powder, 195 mg, 33.2 %). 1 H NMR (500 MHz, DMSO-d6) H (d, J = 8.62Hz, 1H, amido), (d, J = 7.8Hz, 2H, 3-position of stilbene), (d, J = 7.9Hz, 2H, 2-position of stilbene), (d, J = 7.8Hz, 2H, 2 -position of stilbene), (m, J = 7.8Hz, 3H, 3 -position), (d, J = 17Hz, 2H, olefin of stilbene), (t, J = 7.2Hz, 2H, 4 -position), (m, 12H, O2,3H of CD), (m, 6H, C1H of CD), (m, 5H, O6H of CD), (m, overlaps with HOD, others of CD). MALDI TOF-MS, m/z (%), (100) [M+Na]+ Mp oc dec. S7

8 Anal. calced for C 51 H 71 O 30 N 8.9 H 2 O: C,45.74; H,6.76; N,1.05. Found: C,45.44; H,6.45; N,1.11. c) Preparation of mono-3-cis-stilbene amide--cd (3-cis-Sti--CD) 3-trans-sti--CD (100 mg, mmol) was dissolved in methanol (50mL). After the photoirradiation with visible light ( = 340 nm) for an hour, the solution was evaporated. The dried residue was separated by preparative reversed-phase high performance liquid chromatography to give pure 3-cis-sti--CD (white powder, 31 mg, 26.3%). Typical chromatogram of the mixture of two isomers was shown in Figure. The instrumental parameters are given below: Controll system Waters 600 Controler Gradient program Detector Waters 2487 Dual l Absorbancs Detector Time (min) H 2 O / acetonitrile (% v/v) pump Waters Delta 600 initial 75 / 25 Column SunFire C18 OBD mm 11 75/ 25 Recorder Flow rate Temperature Ditection SEKOINIC SS-250F 15ml / min 40 o C (ambient) UV@ 300 nm S8

9 Fraction 1 Fraction 2 Figure. Chromatogram of the mixture of 3-cis-Sti-CD and 3-trans-Sti-CD. Fraction 1 and Fraction 2 were identified as 3-cis-Sti-CD and 3-trans-Sti-CD, respectively, which were characterized by 1 H NMR (Figures S2 and S3). 1 H NMR (500 MHz, DMSO-d 6 ) H (d, J = 8.6Hz,1H, amido), (d, J = 8.32Hz, 2H, 3-position of stilbene), (m, 7H, 2,2 3,4 -position of stilbene), (d, J = 12.4Hz, 2H, olefin of stilbene), (m, 12H, O2,3H of CD), (m, 6H, C1H of CD), (m, 5H, O6H of CD), (m, overlaps with HOD, others of CD). MALDI TOF-MS, m/z (%), (100) [M+Na] +. Mp o C dec. Anal. calced for C 51 H 71 O 30 N 9.9 H 2 O: C:45.16 H: 6.75 N: Found C:44.95 H: 6.52 N: S9

10 Figure S2. The 1 H-NMR spectrum of 3-trans-Sti--CD in DMSO at 30 o C. Figure S3. The 1 H-NMR spectrum of 3-cis-Sti--CD in DMSO at 30 o C. 0

11 Figure S4. The 1 H-NMR spectra of 3-cis-Sti--CDs in D 2 O at 30 o C a) after and b) before irradiation at 254nm. The negative Cotton band around 320 nm was observed in na aqueous solution, corresponding to the 1 La transition band of a stilbene amide group. This Cotton band is assigned to the induced Cotton band between a stilbene amide group and -CD. Figure S5. The circular dichroism spectra of 3- trans-sti--cd in an aqueous solution at 30 o C. (1.0 mm) 1

12 Figure S6. The circular dichroism spectra of 3-cis-Sti--CD in an aqueous solution (black line), 3-cis- Sti--CD in MeOH (red line), and 3-cis-Sti--CD with -CD in an aqueous solution (blue line) at 30 o C. The negative-positive Cotton band around 300 nm of 3-cis-Sti--CD was observed in aqueous solutions, corresponding to the 1 La transition band of the stilbene amide group. This Cotton band is assigned to the exciton coupling interactions between stilbene amide groups. It is noted that two or more excitons are located in relatively close distance. On the contrary, 3-cis-Sti--CD did not exhibit Cotton band in methanol because of the dissociation of these supramolecular assemblies. The mixture of 3-cis-Sti--CD and -CD in an aqueous solution showed the negative Cotton band because the cisstilbene amide group is strongly included in the -CD cavity. These results indicate that 3-cis-Sti--CD formed supramolecular self-assembly by - stacking interactions between stilbene amide groups. 2

13 Figure S7. The 1 H-NMR spectra of various concentrations of 3-cis-Sti--CDs in D 2 O at 30 o C.(0.91 mm, 2.5 mm, 3.1 mm, 6.1 mm, 12 mm, 24 mm, 49 mm, and 98 mm) The protons of 3-cis-Sti--CD showed upfield shifts with an increase in the concentration, indicating the formation of the supramolecular assembly by - stacking interactions in aqueous solutions. Figure S8. Concentration dependence of of 3- cis-sti--cd at 30 o C.(0.91 mm, 2.5 mm, 3.1 mm, 6.1 mm, 12 mm, 24 mm, 49 mm, and 98 mm) 3

14 Figure S9. The 1 H-NMR spectra of 3-cis-Sti--CDs in D 2 O at 30 o C.(5.0 mm) a) before and b) after adding -CD(50 mm) To investigate the formation of supramolecular assembly from 3-cis-Sti--CD, -CD as a competitive host was added to the solution of 3-cis-Sti--CD. After addition of -CD, protons of a stilbene amide group shifted to downfield. This result indicates that the supramolecular assembly was decomposed by the formation of inclusion complexes between -CD and the stilbene amide group. 4

15 Figure 0. Concentration dependence of absorption maximum at 290nm of 3-cis-Sti--CD ( ) in aqueous solutions. ( mm, mm, mm, mm, 0.11 mm, 0.16 mm, 0.24 mm, 0.36 mm, and 0.54 mm) The absorption maxmum of 3-cis-Sti--CD shifted to longer wavelength with an increase in the concentration. This result indicates the formation of supramolecular assembly consisting of 3-cis-Sti-- CD. 5

16 a) 2 g 2 2 ( - /3 - /2) ln (I / I 0 ) b) 2 g 2 2 ( - /3 - /2) ln (I / I 0 ) Caption are shown in Page 18. 6

17 c) 2 g 2 2 ( - /3 - /2) ln (I / I 0 ) d) 2 g 2 2 ( - /3 - /2) ln (I / I 0 ) Caption are shown in Page 18. 7

18 e) 2 g 2 2 ( - /3 - /2) ln (I / I 0 ) Figure 1. Stejskal-Tanner plots ln(i/i 0 ) vs. g 2 of 3-cis-Sti--CD with increased in the concentration of 3-cis-Sti--CD (1 mm (a), 10 mm (b), 25 mm (c), 50 mm (d), and 100 mm (e))at 30 o C in D 2 O. 8

19 Figure 2. Concentration dependence of diffusion coefficient values (D) of supramolecular complexes formed 2-CiO--CD (open square) and 3-cis-Sti--CD (open circle) in D 2 O at 30 C by NMR. To estimate the molecular size of the supramolecular self-assembly from 3-cis-Sti--CD, self-diffusion coefficients (Ds) were determined by the pulsed field gradient spin-echo NMR measurement. We have chosen 2-cinnamoyl--CD (2-CiO--CD) as a reference compound because 2-CiO--CD formed the double threaded dimer. The Ds of 2-CiO--CD showed cm 2 s -1 in the lower concentration region (10 30 mm). The Ds of 3-cis-Sti--CD significantly decreased with an increase in concentration (< 100 mm) (Figure 1). It should be noted that the hydrodynamic radius of 3-cis-Sti-- CD is larger than that of 2-CiO--CD at the whole region of the concentration. These results suggest that the hydrodynamic volume of supramolecular self-assembly from 3-cis-Sti--CD showed larger than that of a double threaded dimer in aqueous solutions. 9

20 Figure 3. Turbo-ion spray mass spectrum of 3-cis-Sti--CD in an aqueous solution. S2 0

21 Table. Crystal data and collection parameters for 3-trans-Sti--CD. 3-trans-Sti--CD formula C 45 H 66 O 31 formula weight crystal system orthorhombic space group P (#96) a, Å (4) b, Å (4) c, Å (6), deg. -, deg. -, deg. - V, Å (4) Z 8 D calcd, g/cm F(000) µ [Cu K], cm temp, K min, 2 max, deg 3.2, 68.2 no. of refl. Measured (Total) 6177 no. of refl. measured (Unique) 4850 no. of variables 704 R wr GOF S2 1