SUPPORTING INFORMATION
A Straightforward Strategy to Substitute Amide Bonds by 1,2,3 Triazoles in peptaibols
using Aib[Tz]-Xaa dipeptides.
Khoubaib Ben Haj Salah,1 Baptiste Legrand,2 Sanjit Das,1 Jean Martinez, 2 Nicolas
Inguimbert1
1
Université de Perpignan Via Domitia, Centre de Recherche Insulaire et Observatoire de
l’Environnement (CRIOBE) USR CNRS 3278, centre de phytopharmacie, batiment T, 58
avenue P. Alduy, 66860 Perpignan, France.
2
Institut des Biomolécules Max Mousseron (IBMM), UMR 5247 CNRS, Université de
Montpellier, 15 avenue Charles Flahault, BP 14491, 34093 Montpellier Cedex 5
1
H and 13C NMR spectra. HPLC chromatograms, ESI Mass spectra.
S1
Table of Contents of the Supporting Information
Title, authors, and address
S1
Table of contents
S2
1
S3
13
H, C NMR of compound 2
Analytical LC-MS of compound 2
S4
1
S5
13
H, C NMR of compound 3a
Analytical HPLC-ELSD and LC-MS of compound 3a
S6
1
S7
13
H, C NMR of compound 3b
Enlargement of 13C spectra for compound 3b
S8
Analytical HPLC-ELSD and LC-MS of compound 3b
S9
1
S10
13
H, C NMR of compound 3c
Analytical HPLC-ELSD and LC-MS of compound 3c
S11
1
S12
H, 13C NMR of compound 3d
Analytical HPLC-ELSD and LC-MS of compound 3d
S13
1
S14
13
H, C NMR of compound 3e
Analytical HPLC-ELSD and LC-MS of compound 3e
S15
1
S16
H, 13C NMR of compound 3f
Analytical HPLC-ELSD and LC-MS of compound 3f
H, C NMR of compound 3f’
1
13
S17
S18
Analytical HPLC-ELSD and LC-MS of compound 3f’
S19
1
S20
Analytical HPLC-ELSD and LC-MS of compound 3g
S21
Analytical HPLC-ELSD and MS of compound 4b
S22
Analytical HPLC-ELSD and MS of compound 4d
S23
Analytical HPLC-ELSD and MS of compound 4e
S24
Analytical HPLC-ELSD and MS of compound 4f
S25
1
S26
H, 13C NMR of compound 3g
13
H, C NMR of compound 4f
Analytical HPLC-ELSD and MS of compound 4f’
H, 13C NMR of compound 4f’
S27
1
S28
Analytical HPLC-ELSD and MS of compound 4g
S29
Analytical HPLC-ELSD and MS of crude compound 6a
S30
Analytical HPLC-ELSD and MS of compound 6b
S31
Analytical HPLC-ELSD coinjection compounds 6a and 6b
S32
Analytical HPLC-ELSD and MS of compound 6c
S33
Analytical HPLC-ELSD and MS of compound 6d
S34
Analytical HPLC-ELSD and MS of compound 7
S35
Table S1 1H NMR chemical shifts for compound 7 and 6d
S36
Table S2 Interresidue NOEs for compound 6d and 7
1
H NMR spectra of compound 7 and 6b in CD3OH at 293 K.
S37
S38
S2
9H-fluoren-9-ylmethyl (2-methylbut-3-yn-2-yl)-carbamate (2)
1
H NMR (400 MHz, CDCl3)
13
C NMR (100 MHz, CDCl3)
S3
Analytical LC-MS gradient: 0.5 ml/min flow rate from 10% B to 100% B over 30 min and 10% B
over 5 min
S4
Fmoc-Aib[Tz]Aib-OH (3a)
1
H NMR (400 MHz, DMSO-d6)
13
C NMR (100 MHz, DMSO-d6)
S5
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
1800.00
1600.00
1400.00
1200.00
LSU
1000.00
800.00
600.00
400.00
200.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
Analytical LC-M gradient: 0.5 ml/min flow rate from 10% B to 100% B over 30 min and 10% B
over 7 min
S6
Fmoc-Aib [Tz]Gln(Trt)-OH (3b)
1
H NMR (400 MHz, DMSO-d6)
13
C NMR (100 MHz, DMSO-d6)
S7
S8
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
1800.00
1600.00
1400.00
1200.00
LSU
1000.00
800.00
600.00
400.00
200.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
Analytical LC-MS gradient: 0.5 ml/min flow rate from 10% B to 100% B over 30 min and 10% B
over 7 min
S9
Fmoc-Aib [Tz]Gly-OH (3c)
1
H NMR (400 MHz, DMSO-d6)
13
C NMR (100 MHz, DMSO-d6)
S10
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
2000.00
1800.00
1600.00
1400.00
LSU
1200.00
1000.00
800.00
600.00
400.00
200.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
Analytical LC-MS gradient: 0.5 ml/min flow rate from 10% B to 100% B over 30 min and 10% B
over 7 min
S11
Fmoc-Aib [Tz]Ala-OH (3d)
1
H NMR (400 MHz, DMSO-d6)
13
C NMR (100 MHz, DMSO-d6)
S12
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
1200.00
1000.00
LSU
800.00
600.00
400.00
200.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
Analytical LC-MS gradient: 0.5 ml/min flow rate from 10% B to 100% B over 30 min and 10% B
over 7 min
S13
Fmoc-Aib [Tz]Leu-OH (3e)
1
H NMR (400 MHz, DMSO-d6)
13
C NMR (100 MHz, DMSO-d6)
S14
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
2000.00
1800.00
1600.00
1400.00
LSU
1200.00
1000.00
800.00
600.00
400.00
200.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
Analytical LC-MS gradient: 0.5 ml/min flow rate from 10% B to 100% B over 30 min and 10% B
over 7 min
S15
Fmoc-Aib [Tz]Val-OH (3f)
1
H NMR (400 MHz, DMSO-d6)
13
C NMR (100 MHz, DMSO-d6)
S16
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
800.00
700.00
600.00
LSU
500.00
400.00
300.00
200.00
100.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
Analytical LC-MS gradient: 0.5 ml/min flow rate from 10% B to 100% B over 30 min and 10% B
over 7 min
S17
Fmoc-Aib [Tz]-D-Val-OH (3f’)
1
H NMR (400 MHz, DMSO-d6)
13C
NMR (100 MHz, DMSO-d6)
S18
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
800.00
700.00
600.00
LSU
500.00
400.00
300.00
200.00
100.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
Analytical LC-MS gradient: 0.5 ml/min flow rate from 10% B to 100% B over 30 min and 10% B
over 7 min
S19
Fmoc-Aib [Tz]Tyr(OMe)-OH (3g)
1H
13C
NMR (400 MHz, DMSO-d6)
NMR (100 MHz, DMSO-d6)
S20
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
1400.00
1200.00
LSU
1000.00
800.00
600.00
400.00
200.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
Analytical LC-MS gradient: 0.5 ml/min flow rate from 10% B to 100% B over 30 min and 10% B
over 7 min
S21
Fmoc-Aib [Tz]Gln(Trt)-Ile-OMe (4b)
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
1100.00
1000.00
900.00
800.00
LSU
700.00
600.00
500.00
400.00
300.00
200.00
100.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
ESI-MS
S22
Fmoc-Aib [Tz]Ala-Ile-OMe (4d)
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
2000.00
1800.00
1600.00
1400.00
LSU
1200.00
1000.00
800.00
600.00
400.00
200.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
ESI-MS
S23
Fmoc-Aib [Tz]Leu-Ile-OMe (4e)
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
2000.00
1800.00
1600.00
1400.00
LSU
1200.00
1000.00
800.00
600.00
400.00
200.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
ESI-MS
S24
Fmoc-Aib [Tz]Val-Ile-OMe (4f)
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
2000.00
1800.00
1600.00
1400.00
LSU
1200.00
1000.00
800.00
600.00
400.00
200.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
ESI-MS
S25
Fmoc-Aib [Tz]Val-Ile-OMe (4f)
1H
13C
NMR (400 MHz, DMSO-d6)
NMR (100 MHz, DMSO-d6)
S26
Fmoc-Aib [Tz]-D-Val-Ile-OMe (4f’)
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
800.00
700.00
600.00
LSU
500.00
400.00
300.00
200.00
100.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
ESI-MS
S27
Fmoc-Aib [Tz]-D-Val-Ile-OMe (4f’)
1H
13
NMR (400 MHz, DMSO-d6)
C NMR (100 MHz, DMSO-d6)
S28
Fmoc-Aib [Tz]Tyr(OMe)-Ile-OMe (4g)
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
1800.00
1600.00
1400.00
LSU
1200.00
1000.00
800.00
600.00
400.00
200.00
0.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
Minutes
28.00
30.00
32.00
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
ESI-MS
S29
Ac- Phe-Aib[Tz]Aib-Aib[Tz]Val-Aib[Tz]Gly-Gly-OH (6a)
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate 10% B for 10 min and to 30 % B over 40
min.
gradient: 0.9 ml/min flow rate 20% B for 5 min and to 30 % B over 40 min.
gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
ESI-MS
S30
Ac- Phe-Aib[Tz]Aib-Aib[Tz]-D-Val-Aib[Tz]Gly-Gly-OH (6b)
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate 10% B for 10 min and to 30 % B over 40
min.
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate 20% B for 5 min and to 30 % B over 40
min.
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
ESI-MS
S31
co-injection of (6a) and (6b)
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate 20% B for 5 min and to 30 % B over 40
min.
S32
Ac-(Aib[Tz]Leu)6-NH2 (6c) Crude
Analytical HPLC-ELSD gradient: 0.9 ml/min flow rate from 10% B to 100% B over 50 min.
ESI-MS
S33
Ac-(Phe-Aib[Tz]Ala)3-NH2 (6d)
Analytical LC-MS gradient: 0.5 ml/min flow rate from 10% B to 100% B over 30 min and 10% B
over 5 min
S34
Ac-(Phe-Aib-Ala)3-NH2 (7)
Analytical LC-MS gradient: 0.5 ml/min flow rate from 10% B to 100% B over 30 min and 10% B
over 5 min
S35
1
H NMR chemical shifts for 7 and 6d in CD3OH
1
H NMR chemical shifts for 7 in CD3OH at 288 K
Residue
Phe 1
Aib 2
Ala 3
Phe 4
Aib 5
Ala 6
Phe 7
Aib 8
Ala 9
HN
8.36
8.41
7.70
8.01
7.93
7.71
7.97
7.77
7.60
H
4.43
1.32
4.39
1.27
4.35
1.44
H
3.08
1.43
3.12, 3.25
1.42
3.07, 3.25
1.42
-
Others
CH3 2.02
NH2 6.93, 7.32
1
H NMR chemical shifts for 6d in CD3OH at 288 K
Residue
Phe 1
Aib 2
Ala 3
Phe 4
Aib 5
Ala 6
Phe 7
Aib 8
Ala 9
HN
8.05
8.07
8.29
8.13
8.29
8.17
-
H
4.58
1.69
4.54
1.72
4.55
1.76
H
2.88, 3.02
1.53, 1.63
2.94, 3.04
1.54, 1.65
2.95, 3.05
1.54, 1.66
-
Others
CH3 2.02
HψTz 7.73
HTz 7.80
HψTz 7.84
NH2 7.27, 7.70
S36
Unambiguous inter-residue NOEs correlations observed in the ROESY spectrum of 7 and 6d
in CD3OH at 293 K. Strong “s” < 2.7 Å; medium “m” < 3.3 Å; weak “w” < 5.0 Å.
Name of the NOE
correlation
Ac-HN1
HN2-HA1
HN2-HB1
HN2-HN3/HTz(2-3)
HN3/HTz(3-4)-HN4
HN3/HB2
HN3/HTz(3-4)-HA1
HN4-HA3
HN4-HB3
HN4-HB2
HN5-HN6/HTz(5-6)
HN5-HA4
HN5-HB4
HN5-HA3
HN6/HTz(5-6)-HA4
HN6/HTz(5-6)-HB5
HN6/HTz(5-6)-HN7
HN7-HN8
HN7-HA6
HN7-HB6
HN8-HN9/HTz(8-9)
HN8-HA7
HN8-HB7
HN8-HA6
HN9/HTz(8-9)-HB8
HN9/HTz(8-9)-HA7
NOE
7
m
m
w
w
w
w
m
w
w
m
w
w
w
w
w
w
w
w
w
w
w
w
6d
m
s
w
w
s
m
w
s
w
w
s
m
w
s
w
w
Type
Sequential
Sequential
Sequential
Sequential
Sequential
Sequential
Medium
Sequential
Sequential
Medium
Sequential
Sequential
Sequential
Medium
Medium
Sequential
Sequential
Sequential
Sequential
Sequential
Sequential
Sequential
Sequential
Medium
Sequential
Medium
S37
1
H NMR spectra of compound 7 and 6b in CD3OH at 293 K.
S38