Scaffolded Multiple Cyclic Peptide libraries for Protein Mimics by Native Chemical Ligation

H. van de Langemheen,a M. van Hoeke,a H (Linda). C. Quarles van Ufford,a J. A. W. Kruijtzera and R. M. J. Liskampa,b

a Medicinal Chemistry & Chemical Biology, Utrecht Institute for Pharmaceutical Sciences, Department of Pharmaceutical Sciences, Faculty of Sciences, Utrecht

University, P.O. Box 80082, 3508 TB, Utrecht, The Netherlands.

b School of Chemistry, Joseph Black Building, University of Glasgow, University Avenue, Glasgow G12 8QQ, United Kingdom.

R.M.J.Liskamp@uu.nlRobert.Liskamp@glasgow.ac.uk

1

Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2014

(Boc-Cys(Trt))3-TAC(N(H)Me) 4

1H NMR at 373 K

13C NMR at 373 K

2

HSQC at 373 K

COSY at 373 K

3

(H-Cys)3-TAC(N(H)Me) 5

HPLC

Minutes0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0

Vol

ts

0.00

0.25

0.50

0.75

1.00

1.25

1.50

1.75

2.00

Cyclic thioester loop 1 (11a)

HPLC

Minutes0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32

Vol

ts

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

4

MALDI-TOF

0

10

20

30

40

50

60

70

80

90

100

%Int.

1020 1022 1024 1026 1028 1030 1032 1034 1036 1038 1040 1042 1044 1046 1048 1050 1052 1054 1056 1058 1060 1062 1064 1066 1068 1070 1072m/z

1[c].H22

473 mV[sum= 21270 mV] Profiles 1-45 Unsmoothed -Baseline 60

Data: loop1cyano0001.H22[c] 4 Jul 2012 15:38 Cal: P14R120704 4 Jul 2012 15:29 Shimadzu Biotech Axima CFR 2.9.3.20110624: Mode Reflectron, Power: 25, P.Ext. @ 1500 (bin 120)

1030.402

1031.409

1032.418

1052.3861033.425 1068.346

1054.4051039.8331031.831 1044.4721036.222

Cyclic thioester loop 2 (11b)

HPLC

Minutes0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32

Vol

ts

0.0

0.5

1.0

1.5

2.0

2.5

MALDI-TOF

0

10

20

30

40

50

60

70

80

90

100

%Int.

1463 1464 1465 1466 1467 1468 1469 1470 1471 1472 1473 1474 1475 1476 1477 1478 1479 1480 1481 1482 1483 1484 1485 1486m/z

1[c].H21

154 mV[sum= 8301 mV] Profiles 1-54 Unsmoothed -Baseline 60

Data: loop2cyano20001.H21[c] 4 Jul 2012 15:46 Cal: P14R120704 4 Jul 2012 15:29 Shimadzu Biotech Axima CFR 2.9.3.20110624: Mode Reflectron, Power: 30, P.Ext. @ 1500 (bin 120)

1473.607

1474.608

1475.627

1476.603

1473.028 1476.097 1477.641 1482.6991479.2361466.1441463.949 1468.474 1484.368

5

Cyclic thioester loop 3 (11c)

HPLC

Minutes0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32

Vol

ts

0.0

0.2

0.4

0.6

0.8

1.0

MALDI-TOF

0

10

20

30

40

50

60

70

80

90

100

%Int.

1170 1172 1174 1176 1178 1180 1182 1184 1186 1188 1190 1192 1194 1196 1198 1200 1202 1204 1206 1208 1210 1212 1214m/z

1[c].H20

156 mV[sum= 4985 mV] Profiles 17-48 Unsmoothed -Baseline 60

Data: loop3cyano0001.H20[c] 4 Jul 2012 15:32 Cal: P14R120704 4 Jul 2012 15:29 Shimadzu Biotech Axima CFR 2.9.3.20110624: Mode Reflectron, Power: 32, P.Ext. @ 1500 (bin 120)

1194.608

1195.591

1172.594

1173.784

1210.555

1196.630

1212.5441175.612 1197.838 1202.663 1209.3901178.721 1189.5061171.417 1192.4141185.9361182.142 1214.0391206.373

6

Triple-ligated product 12

HPLC

Minutes0 5 10 15 20 25 30 35 40 45 50

Vol

ts

0.0

0.5

1.0

1.5

2.0

2.5

MALDI-TOF

0

10

20

30

40

50

60

70

80

90

100

%Int.

3270 3275 3280 3285 3290 3295 3300 3305 3310m/z

1[c].N2

136 mV[sum= 3955 mV] Profiles 1-29 Unsmoothed -Baseline 60

Data: sinTACloop1io0002.N2[c] 28 Mar 2012 11:19 Cal: 120328IO 28 Mar 2012 11:27 Shimadzu Biotech Axima CFR 2.9.3.20110624: Mode Reflectron, Power: 40, P.Ext. @ 3285 (bin 178)

3287.46

3289.37

3286.493290.37

3291.37

3274.783293.38 3302.553284.583269.50 3279.82

7

Mixture 13

Mass

Triple-capped product 13 – [1-1-1] mimic

HPLC

Minutes0 5 10 15 20 25 30 35 40 45 50

Vol

ts

-0.1

0.0

0.1

0.2

0.3

0.4

0.5

0.6

8

mass

[1-1-3] mimic

HPLC

Minutes0 5 10 15 20 25 30 35 40 45 50

Vol

ts

-0.1

0.0

0.1

0.2

0.3

0.4

9

mass

[1-3-3] mimic

HPLC

Minutes0 5 10 15 20 25 30 35 40 45 50

Vol

ts

-0.100

-0.075

-0.050

-0.025

0.000

0.025

0.050

0.075

0.100

10

mass

[1-1-2] mimic

HPLC

Minutes0 5 10 15 20 25 30 35 40 45 50

Vol

ts

-0.10

-0.05

0.00

0.05

0.10

0.15

11

mass

[1-2-3] mimic

HPLC

Minutes0 5 10 15 20 25 30 35 40 45 50

Vol

ts

-0.1

0.0

0.1

0.2

0.3

0.4

0.5

0.6

12

mass

[2-3-3] mimic

HPLC

Minutes0 5 10 15 20 25 30 35 40 45 50

Vol

ts

-0.10

-0.05

0.00

0.05

0.10

0.15

13

mass

[1-2-2] mimic

HPLC

Minutes0 5 10 15 20 25 30 35 40 45 50

Vol

ts

-0.1

0.0

0.1

0.2

0.3

0.4

14

mass

[2-2-3] mimic

HPLC

Minutes0 5 10 15 20 25 30 35 40 45 50

Vol

ts

-0.1

0.0

0.1

0.2

0.3

0.4

mass

15

Triple-capped TAC scaffold (for HIV-1 gp120 capture ELISA)

HPLC

Minutes0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0

Vol

ts

-0.1

0.0

0.1

0.2

0.3

0.4

16