ch cn and 0.1% trifluoroacetic acid (tfa) and buffer b (5m ...microwave assisted peptoid syntheses...

Supplemental Information

Solid-Phase Synthesis of Peptoid-like Oligomers Containing Diverse Diketopiperazine Units

Sujit Suwal and Thomas Kodadek*

Materials and Instruments:Amino acid esters were purchased from either from Aldrich, Acros, Advanced chemtech and Astatech. (S) and (R)-2-bromo-propionic acid were purchased from Aldrich. Mono-N-tboc-ethylenediamine was bought from Astatech Inc. Primay amines were purchased from Advanced Chemtech, Aldrich, Acros, TCI. Rest of the other chemicals and reagents were purchased either from Aldrich or Fisher Scientific. Rink amide MBHA resin (0.5 mmol/g) and Tentagel macrobead-NH2 (160 µg, 0.4 mmol/g) resin were obtained from Chemprep and rapp-polymere, respectively. Disposable fritted columns (5 mL, 50 mL) for peptoid syntheses were bought from Intavis AG. Microwave assisted peptoid syntheses were accomplished utilizing 10% of 1550 W household microwave (GE model JE 1860 BH04). HPLC purification of crude peptoids after acid cleavage from Rink amide resin was carried out in Water 1525 binary HPLC pumps and a 2487 dual absorbance detector, or a 2998 photodiode array detector. Buffer A (H2O with 5% CH3CN and 0.1% trifluoroacetic acid (TFA) and buffer B (5m, 250 x 4.6 mm, Alltech) and Apollo C-18 5µ preparative column. Purities of the compounds were assessed by analytical HPLC under UV (214), otherwise mentioned in description of the spectrum. MS and MS/MS were recorded in MALDI -TOF (4800 Proteomics Analyser, Applied Biosystems) or ESI-MS (Waters). In MS/MS, the signal-to-noise ratio threshold set to 10. Cyano-4-hydroxycinnamic acid (HCCA) or charcoal was used as MALDI matrices (Aldrich). 1H and 13C NMR spectra were recorded in Bruker 400 instrument operating at 400 MHz for 1H and 100 MHz for 13C NMR using Deuterated-methanol (CD3OD) or Deuterated-Dimethylsulfoxide (DMSO) (Cambridge) as a solvent.

Synthesis of 2, 5-DKP (4 a-f)

HN

Br

O

HN

NH

O

O

OR"R

HN

N

O

O

OR"R

O

NH

H2NN

O

O

NR

O

R'

Scheme 1Amino acid esterDMF, 37oC, ON

1. BAA, DIC 37oC, 2x 30 min2. Primary amine DMF, 37oC, 1 h

1. 50 - 60oC, ON

R'

1 2 3

4 a-f

Where,

2. TFA:DCM:TIS (60:38:2)

a. R = (S)-Me, R' = Benzyl

e. R = (S)-Me, R' =OH

b. R = (S)-4-hydroxybenzyl, R'= 2- methoxy ethylamine

R"= CH3 or C2H5

c. R = (S)-Me, R' = Ph

d. R = (R)-Me, R' = Ph f. R = (R)-Me, R' = OH

1

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

N

N

O

O

H2N

O

4a

Rink amide MBHA resin (1 g, 0.7 mmol) was swelled in DMF. Fmoc group was deprotected with 20% piperidine in DMF (2X, 15 mins). The resultant primary amine was coupled with bromoacetic acid (BAA, 2M) activated with diisopropylcarbodiimide (DIC, 2 M) in N,N’-dimethylformamide (DMF). Subsequently, the bromide was displaced with desired HCl salt of amine acid ester in DMF (2 M) along with DIEA (1.5X with respect to ester). The secondary amine was further coupled with bromoacetic acid (2M), activated with DIC (3M), 37°C (2X, 15 min). The bromide was displaced with a solution of desired primary amine (viz benzyl amine in this case) in DMF at 37oC, 1 h. At this stage, the reaction mixture further incubated at 50oC overnight so that the secondary amine thus generated after SN2 reaction undergo addition elimination at the ester thus generating six-membered 2,5 diketopiperazine. Finally, the solid support was washed with DMF, DCM and ethanol, and subjected to cleave with 50% TFA in DCM along with 2% triisopropylsilane. A crude DKP obtained after the solvent was evaporated under Argon and ether precipitation. A crude dried compound finally was subjected to NMR study without further HPLC purification by dissolving in d-methanol. 1HNMR (400 MHZ, D6-DMSO) δ 7.4-7.3 (m, 5H), 4.7 (d, 1H, J=14.7), 4.5 (d, 1H, J=14.7), 4.3 (d, 1H, J=16.7), 4.12 (q, 1H), 4.0 (d, 1H, J=17.7), 3.9 (d, 1H, J=16.7), 3.82 (d, 1H, J= 17.5), 1.5 (d, 3H, Me, J=7.2); 13CNMR (100 MHz, DMSO) δ 172.3, 169.2, 166.5, 136.8, 129.9, 59.19, 50.3, 49.7, 47.8, 18.1; MALDI-TOF MS (Obs.=MNa+) 298.0681 (298.1270 expected).

Figure S1: 1HNMR of compound 4a

2

Figure S2: COSY of 4a

Figure S3: 13CNMR of 4a

Figure S4: Dept of 4a

3

N

N

O

O

H2N

O

Exact Mass: 275.13

Fig S5: MALDI-MS of compound 4a

N

N

O

O

H2N

O

4bOH

O

Spectral data for compound 4b:A crude dried compound was dissolved in in d-DMSO. 1HNMR (400 MHZ, D6-DMSO) δ 6.83 (d, 2H, J=8.3), 6.66 (d, 2H, J=8.3) 4.35 (d, 1H, J=16.4), 4.15 (t, 1H), 3.6 (d, 1H, J=16.4), 3.32 (m, 1H, J=17.1), 3.5 (d, 1H), 3.35 (m, 1H), 3.3 (m, 1H), 3.2 (s, 3H), 3.15 (m, 1H), 3.0 (dd, 1H, J=14), 2.9 (dd, 1H, J=14), 2.6 (d, 1H, J=17.1); 13CNMR (100 MHz, DMSO) δ 169.2, 165.3, 164.0, 156.5, 130.7, 124.9, 115.0, 69.2, 61.7, 57.8, 49.5, 45.8, 44.9, 35.5; ESI-MS(Obs. MNa+) =358.1398 (Expected=358.1481).

Figure S6: 1HNMR of 4b

4

Figure S7: 13CNMR of compound 4b

Figure S8: 13C-DEPT of compound 4b

Figure S9: COSY of compound 4b

5

N

N

O

O

H2N

O

OH

O

Exact Mass: 335.15

Fig S10: MALDI-MS of compound 4b

Spectral data for compound 4c and 4d:

For compound 4d:

A HPLC purified compound was dissolved in in d-chloroform. 1HNMR (400 MHZ) δ 7.3-7.2 (m, 5H), 4.03-3.97 (m, 2H), 3.90 (d, 1H), 3.60 (d, 1H), 3.35 (d, 1H), 3.23 (m, 1H), 3.02 (m, 1H), 1.45 (d, 3H), 1.29 (d, 3H); 13CNMR (100 MHz, DMSO) δ 172.6,169.6, 166.9, 63.1, 59.2, 52.3, 47.7, 45.5, 17.7, 13.4; MALDI-TOF MS (Obs. MNa+) 326.0962 (Expected = 326.1304 expected).

For compound 4c:

A HPLC purified compound was dissolved in in d-chloroform. 1HNMR (400 MHZ) δ 7.3-7.2 (m, 5H), 4.0 (d, 1H), 3.95-3.84 (m, 3H), 3.83 (d, 1H), 3.61 (d, 1H), 3.40-3.35 (m, 1H), 3.15-310 (m, 1H), 1.3 (d, 3H), 1.2 (d, 3H); MALDI-TOF MS (Obs. MNa+) 326.0962 (Expected = 326.1304 expected).

6

N

N

H2N

O

4d

O

O

Me

Meb

c

N

N

H2N

O

4c

O

O

Me

Meb

a

Figure S11: 1HNMR of compound 4c and 4d

N

N

H2N

O

4d

O

O

Me

Meb

c

N

N

H2N

O

4c

O

O

Me

Meb

a

Figure S12: 13CNMR of compound 4c and 4d

7

N

N

H2N

O

4d

O

O

Me

Meb

c

N

N

H2N

O

4c

O

O

Me

Meb

a


N

N

H2N

O

4c

O

O

Me

Meb

a

N

N

H2N

O

4c

O

O

Me

Meb

a


8

N

N

H2N

O

4d

O

O

Me

Meb

c

N

N

H2N

O

4c

O

O

Me

Meb

a

Figure S15: HPLC and ESI-MS of compound 4c and 4d

N

N

O

O

H2N

O

4eOH

Spectral data for compound 4e:A crude dried compound was dissolved in in d4-methanol. 1HNMR (400 MHZ, D4- MeOH) 4.6 (m, 1H), 4.3 (d, 1H), 4.14 (d, 1H), 4.06 (q, 1H), 4.02 (d, 1H), 3.9 (d, 1H), 3.64 (m, 1H), 1.49 (d, 1H), 1.18 (d, 1H); 13CNMR (100 MHz, DMSO) δ 172.6,169.6, 166.9, 63.1, 59.2, 52.3, 47.7, 45.5, 17.7, 13.4; MALDI-TOF MS (Obs. MNa+) 266.0499 (Expected =266.1219).

Figure S16: 1HNMR of compound 4e

9

Figure S17: Cosy of compound 4e

Figure S18: 13CNMR of compound 4e

Fig S19: MALDI-MS of compound 4e

10

N

N

O

O

H2N

O

4fOH

Spectral data for compound 4f:A crude dried compound was dissolved in D6-DMSO. 1HNMR (400 MHZ, D6-DMSO) 4.6 (m, 1H), 4.3 (d, 1H), 4.14 (d, 1H), 4.06 (q, 1H), 4.02 (d, 1H), 3.9 (d, 1H), 3.64 (m, 1H), 1.49 (d, 1H), 1.18 (d, 1H); 13CNMR (100 MHz, DMSO) δ 169.3, 166.8, 164.2, 61.6, 59.7, 43.8, 40.0, 45.5, 17.2, 13.1; MALDI-TOF MS (Obs. MNa+) 266.0122 (266.1219 expected).

Figure S20: 1H NMR of compound 4f

11

Figure S21: COSY of compound 4f

Figure S22: 13C NMR of compound 4f

12

Figure S23: DEPT of compound 4f

Figure S24: MALDI-TOF MS of compound 4f

Figure S25: HPLC of compound 4f

13

Solid phase synthesis protocol for 2,5-DKP containing analog utilized for purity test:

R' =

NNH

O

O

O

NH

1. BAA, DIC, DMF, 37oC, O.N.2. Amino acid ester, DMF, 37-50oC

NN

NH

O

O

O

OR'

O

O

NH H2N

NN

NO

O

O

OR'

N

O

O

1. BAA, DIC, DMF 30 min, 37oC

2. 50-60oC, O.N.3. TFA:DCM:TIS (60:38:2), 2 h.

Ph

NH

N

OH

NH

NH2

H , , ,, ,

, , ,

5 6 7

a b c d e

f g h i

Cl

Cl

Cl

Cl

OScheme 2

Cl

Cl

Rink amide MBHA resin (100 mg, 0.07 mmol) was swelled in DMF. After the fmoc group deprotection with 20% piperidine in DMF (2X, 15 mins), a dimer peptoid was synthesized by conventional peptoid synthesis protocol. Onto the resultant dimer additional bromoacetic acid was added and bromide was displaced by amino acid ester. The secondary amine was further coupled with DIC activated bromoacetic acid at 37°C (2X, 15 min). The bromide was displaced primary amine (2 M) in DMF at 37oC, 1 h. To afford DKP, the reaction mixture was further incubated at 50oC overnight. Completion of the reaction was confirmed by Chloranil test. The compound thus obtained was further attempted for chain extension with DIC activated BAA followed by displacement with primary amine. At this stage, the bead was acid-cleaved and subjected for HPLC to test the purity of DKP formation. HPLC fractions were subjected for MALDI-TOF analysis to confirm the presence of desired molecule.

DKP-Gly:

Figure S26: HPLC of 7a showing retention time at 5 min. with MALDI-TOF (in the middle) showing the molecular mass after the purification.

14

DKP-Ala:

Figure S27: HPLC of 7b showing retention time at 5 min. with MALDI-TOF (in the middle) showing the molecular mass after the purification.

DKP-Ileu:

Figure S28: HPLC of 7c showing retention time at 5 min. with MALDI-TOF (in the middle) showing the molecular mass after the purification.

DKP-Leu:

Figure S29: HPLC of 7d showing retention time at 5 min. with MALDI-TOF (in the middle) showing the molecular mass after the purification.

15

DKP-PhGly:

Figure S30: HPLC of 7e showing retention time at 35 min. with MALDI-TOF (in the middle) showing the molecular mass after the purification.

DKP-Tyr:

Figure S31: HPLC of 7f showing retention time at 5 min. with MALDI-TOF (in the middle) showing the molecular mass after the purification.

DKP-His:

Figure S32: HPLC of 7g showing retention time at 26 min. with MALDI-TOF (in the middle) showing the molecular mass after the purification.

16

DKP-Trp

Figure S33: HPLC of 7h showing retention time at 35 min. with MALDI-TOF (in the middle) showing the molecular mass after the purification

DKP-Lys

Figure S34: HPLC of 7i showing retention time at 26 min. with MALDI-TOF (in the middle) showing the molecular mass after the purification.

HPLC and MALD-TOF with MS/MS data of few compounds that are used to test the feasibility and purity for chain extension purpose. In these particular examples alloc group was utilized as a protecting group instead of tboc- group. The alloc-group was deprotected as explained in our previous paper (Org. Biomol. Chem., 2013, 11, 2088-209).

NH

NN

NN

N

O

O

O

O

O N

O

O

HN

O

N

O

NH

O

H2N

O

NH

S

N

N

Exact Mass: 1258.54

HS 378 / 882

531 / 729

857 / 403

1094 / 167

Figure S35: Top: MALDI-MS data showing parent peak (left) of peptoid (right). Bottom: MALDI-TOF MS/MS of the compound.

17

NH

NN

NN

N

O

O

O

O

O N

O

O

HN

O

N

OHN

O

H2N

HS

O

S

N

N

HN

ONO

HNN

SN

NHO

Exact Mass: 1423.63

378/ 1047

520/ 905

167/ 1259

1106/ 319

319/ 1106

1259/ 167


NH

NN

NN

O

O

O

OO

H2N

O

Exact Mass: 1301.52

HSO

N

HN

N

O

N

N

O

Cl

ClO

O

O

NH

HN

NH

O

HN O

378 / 925

520 / 783 1049 / 254

873 / 430

1102 / 201


18

NH

NN

NN

O

O

O

OO

H2N

O

HSO

N

HN

N

O

N

N

O

O

O

O

NHN

378 / 853

O

OO

Exact Mass: 1229.65

569/ 662

947 / 284 1060/ 171

952 / 279


HPLC and MALD-TOF with MS/MS data of few compounds that resembles DKP-hybrid compound library.

H2N

O

N

O

N

N

O

O

Ph

CF3

O

N

O

CF3

N

Exact Mass: 948.33

CF3

583 / 388

368 / 626

254 / 739

Figure S39: Top: MALDI-MS data showing parent peak (left) of peptoid (right). Bottom: MALDI-TOF MS/MS of the compound

19

NH2O

N

O

N

NO

O

O

O

N

NH2

CF3

O

CF3

O

O

N

O

Exact Mass: 1035.36

675 / 384

484 / 598

388 / 693

924 / 135


H2N

O

N

O

N

N

O

O

CF3

O

N

N

O

N

CF3

OO

H2N

Exact Mass: 946.38

610/ 384

395/ 599

252/ 709


Protocol for peptoid library construction:

DKP-Backbone library:

Tentagel MB NH2 (0.5 g, initial loading 0.4 mmol/g) was soaked in DMF for 30 min, RT. The beads were coupled with fmoc-methionine (5X) activated with N,N,N′,N′-Tetramethyl-O-(1H- benzotriazol-1-yl)uronium hexafluorophosphate (HBTU) (5X), N-Hydroxybenzotriazole (HOBt) (8X) and Diisopropyl ethylamine (DIEA) (10x) in DMF. Mixture was incubated 3 h at room temperature. Fmoc-group was deprotected using 20% piperidine in DMF. Resultant primary amine was treated with BAA/DIC and the bromide was displaced with methoxyethylamine under microwave condition. Likewise, methylamine was added as additional two monomers to represent a linker region. The

20

main chain 2,5-DKP containing peptoid oligomer library (~ 40 K diversity) was created utilizing 12 different commercially available primary amines along with 8 different 2,5-DKP moieties. As a first unit of peptoid library, BAA/DIC was coupled to the secondary amine from the linker. The solid support was split into 12 different fractions after the beads were washed with DMF (5X) and DCM (5X). Each portion of the beads was separately treated with different primary amines (2M) to displace the bromide under microwave condition. The beads were washed as explained above. Resultant secondary amines were pooled and coupled with BAA/DIC. The beads were washed and split into 8 different vessels and treated them separately with amino acid esters. After additional round of BAA/DIC coupling, tboc-protected ethylenediamine was used to displace the bromide. The reaction mixture was incubated 50-60oC, 24h to afford 2,5-DKP formation. T-boc group was deprotected by 50% TFA in DCM. The resultant primary amine was separated into two halves and coupled with DIC-activated BAA or (S)-2-bromopropionic acid separately. The bromides were pooled and split into 12 different fractions and displaced individually by primary amines (R’). Finally a peptoid oligomer library with 2,5-DKP in the main-chain was finally completed by final round to BAA/DIC coupling and bromide displacement with different sets of 12 primary amines (R”, Scheme Sch2).

NN

NNH

O

O

O

O

O

N

R

O HN

O

N

R

O

NH

R"

O

O

A

R'

H2N H2N

O

H2N

NH

O

H2N

H2N

NH2

CF3

H2N

N

O

NH2

O

O

NH2

Ph

H2N

SNH2

O O

SH2N NH2

F

R :

H2N

NH2

H2N

HN

O

H2N

NH2

CF3

H2N

N

O

NH2

O

O

NH2

Ph

H2N

SNH2

O O

SH2N

H2N

NH

N

H2N

OH

H2N

NH

R"

N

N

O

O

N

N

O

O

N

N

O

O

N

N

O

O

N

N

O

O

H2N

N

N

O

O

N

HN

N

N

O

O

NH

N

N

O

O

OH

A

DKP-Side-chain library:

The linker region and the first diversity unit in DKP-side-chain library were created as in the case of DKP-backbone library. After the first diversity element, the chain extension was carried out coupling with BAA/DIC followed by mono-N-tboc-ethylenediamine, as a later point of chain-extension as a side-chain. The secondary amine on the other hand was coupled with BAA and the solid support was split into 5 different reaction vessels. Each vessel was separately treated with amino acid ester. Beads were pooled and split into 8

21

fractions and treated individually with 8 different primary amines. At this stage the reaction mixture was subjected for 2,5-DKP ring formation under basic thermal condition as explained above. Once the cyclization was completed, the t-boc group was deprotected utilizing 50% TFA in DCM. Two additional peptoid monomers were added from the resultant primary amine utilizing conventional peptoid synthesis method via split and pool method.

H2N H2N

O

H2NH2N

NH2

CF3

H2N

N

O

NH2

Ph

H2N

SNH2

O O

SH2N NH2

F

R :

H2N

Cl Cl

O

O

NH2

H2N

Cl Cl

H2N

O

H2N

HN

O

H2N

NH2

CF3

H2N

N

O

H2N

SNH2

O O

SH2N

H2N

NH

N

H2N

OH

H2N

NH

R"

H2N

N

N

O

O

N

N

O

O

N

N

O

O

N

HN

N

N

O

O

N

N

O

O

OH

A

NN

NNH

O

O

O

O

O

N

R

O

AN

HN

O

R'

O

N

R

O

NH

R

HN

S

O

DKP-terminated hybrid peptidomimetic library:The linker region and the first diversity unit in DKP-side-chain library were created as in the case of DKP-backbone library. The solid support was divided into 16 different vessels. In order to incorporate variety of diversity element at the second diversity element, following processes were executed,

1. Out of 16, two of the fractions were separately coupled with DIC activated (S)-2-bromopropionic acid and separately treated with either mono-N-tboc-piperazine or (S)-4-N-Boc-2-methylpiperazine at 50oC, O.N. The t-boc group was deprotected using 50% TFA in DCM prior to third diversity element was incorporated.

2. One portion out of remaining fractions was coupled with BAA/DIC and bromide was displaced with N-mono-t-boc-ethylenediamine. The secondary amine was coupled with DIC activated (S)-2-bromo-3-phenyl-propionic acid. t-boc group was acid-deprotected and the reaction mixture was incubated under 10% DIEA overnight to afford cyclization to afford 2-oxopiperazine unit.

3. Remaining portion of the beads were divided into two fractions after pooling. Each

22

fraction was equilibrated in DCM and treated directly either with 3-chloromethyl benzoyl chloride (10X) or with 4-bromomethyl phenylsulfonyl chloride in DCM (5 mL). A solution of triethylamine (12X) in DCM (5 mL) was slowly added in the acid chloride solution at 0oC. The reaction mixture was brought into the room temperature and allowed to stand for 3 hr. Each vessel was separately washed with DCM several times, pooled together and split into 13 different fractions. The halides were incubated with 13 different amines (R’) separately at 37oC, 3 hr.

After completion of above explained syntheses, all 16 vessels were pooled and coupled with BAA/DIC. The beads were further split into 13 portion and treated with 13 primary amines separately (R’), 37oC, 1 h. Finally, library synthesis was accomplished by incorporating 2,5-DKP as a capping group. The formation of DKP was performed as explained above. At the terminal position, 56 different 2,5-DKPs as capping groups were generated with 7 different amino acid esters and 8 different primary amines.

23

N

N

O

O

RR"

O

DKP:

R= Side-chain from alanine, leucine,lysine, phenylglycine, phenylalanine, histidine, tryptophan

NH

O

DKPN

R'

O

A

Linker

O

NR'

NH

Linker

O

NH

R'

NH

Linker

O

N

R'

O

N

NH

NH

Linker

O

N

R'

O

N

NH

NH

Linker

O

N

R'

O NHR'

NH

Linker

O

N

R'

SO

NH

O

R'

NH

Linker

O

N

R'

O

N

NH

O Ph

1. (S)BPA, DIC, DMF

2. (S)Me-Piperazine, DMF

1. (S)BPA, DIC, DMF

2. Piperazine, DMF

Cl

O

Cl1.

DIEA, DCM, 0oC

2. R'-NH2 , DMF 50oC, 2 h

SCl

O1.

DIEA, DCM, 0oC

2. R'-NH2 , DMF 37oC, 30 min

Br

O1. BAA, DIC, DMF, 37oC, 15 min2. N-tboc-ethylenediamine DMF, 37oC, 30 min

HO

O

Br

Ph

DMF, 37oC, 2 h

3.

4. 50% TFA in DCM, RT, 30 min

5. 10% DIEA in DMF 2 hr, 37oC

a. b. c. d. e.

a.

b.

c.

d.

e.

1. BAA, DIC, DMF

2. R'NH2, DMF

3. DKP formation as shown in Scheme 1

DKP-hybrid library

O

N

O

O

N

O

NLinker =

24

NH2

Cl

NNH2

O

Cl

HN

NH2

O

NH2

SH2N

O O

NH2

F3C

NH2

O

NH2NH2 ONH2

NH2

OO

NH2O

F3C

NH2

NH2

S

R'

NH2

Cl

N

NH2

O

Cl

HN

NH2

O

NH2

SNH2

O

ONH2

CF3

NH2

O

NH2

NH2

O

O

R"

N

N

O

O

RR"

O

DKP:

R= Side-chain from alanine, leucine,lysine, phenylglycine, phenylalanine, histidine, tryptophan

Following are the structures of peptoids from DKP-backbone library, identifed after CNBr mediated cleavage from the resins and subsequent MALDI – MS and MS/MS analyses. Each figure has a peptoid structure showing the mass of corresponding y and b fragments at particular substituent observed during Collision-Induced Dissociation (CID) of the parent peak:

N

N

O

O

O

NN

O

O

HN

O

N

O

NH

NN

O

NH

OO

O

HN O SH2N O

O

O

O

Exact Mass: 1170.5016

501 / 671

712 / 460

966 / 206

359 / 813

O


25

N

N

O

O

O

NN

O

O

HN

O

N

NN

O

NH

OO

O

O

OS

H2NO

O

O

NH


OH

564/ 643

775 / 432

1029 / 178

359 / 848

O


N

N

O

O

O

NN

O

O

HN

O

N

NN

O

NH

OO

O

O

NH


NH2

Cl

Cl

359 / 656

430 / 583

683 / 330

793 / 230

288 / 727O


26

N

N

O

O

O

NN

O

O

HN

O

N

NN

O

NH

OO

O

O

NH

359 / 795

983 / 131

N

O

CF3

NHO


501 / 653

754 / 400

O


N

N

O

O

O

NN

O

O

HN

O

N

NN

O

NH

OO

O

O

NH


HN O

F

359 / 692

456 / 595

729 / 322

885 / 166O


27

N

N

O

O

O

N

O

NH

O

N

NN

O

NH

OO

O

O

NH

SO

H2NO

Ph


329/ 737

632 / 464

855 / 240

O


N

N

O

O

O

NN

O

O

HN

O

N

O

NH

NN

O

NH

OO

O

NHN

F3CS


NH

359 / 860

512 / 707

838 / 381

1053 / 166

O


28

N

N

O

O

O

NN

O

O

HN

O

N

O

NH

NN

O

NH

OO

O

NHN

Ph


S

512 / 628

751 / 389

974 / 167O


N

N

O

O

O

NN

O

O

HN

O

N

O

NH

NN

O

NH

OO

O

NHN

CF3

CF3

NH

N

359 / 887

574 / 672

851 / 395

1080 / 166

288 / 958


O


29

N

N

O

O

O

NN

O

O

HN

O

N

NN

O

NH

OO

O

O

NH

S O

OH2N

OH

NH2


359 / 715

599 / 475

902 / 172

1002 / 72

288 / 786O


N

N

O

O

O

NN

O

O

HN

O

N

NN

O

NH

OO

O

O

NH

S

359 / 757

733 / 383

900 / 175


CF3OH

430 / 686O


\

30

N

N

O

O

O

NN

O

O

HN

O

N

O

NH

NN

O

NH

OO

O

NHN

NH

O

N O

HN

HN

N

S

S


O

1074 / 333

1242 / 166

474 / 933

O


N

N

O

O

O

NN

O

O

HN

O

N

O

NH

NN

O

NH

OO

O

NHN

S

NH

O

N O

HN

HN

NS

456 / 933

1223 / 166

1056 / 333


O


31

NN

NNH

O

O

O

O

O

NO

O

O

N

N

O

O

OH

HN

O

N

O

NH

HN O

O

O

Exact Mass: 1093.51

359 / 736

501 / 594

804 / 291

1023 / 72

288 / 807


N

N

O

O

O

NN

O

O

HN

O

N

NN

O

NH

OO

O

O

NH

F

NH

S

524 / 627

359 / 792

795 / 354

950/ / 201


O


32

N

N

O

O

O

NN

O

O

HN

O

N

O

NH

NN

O

NH

OO

O

NHN

S O

OH2N

NH

O

NO

NH

HN

N

S

OO

NH2


O

358 / 1150

428 / 1079

1343 / 166

1102 / 406


DKP-side chain library:

NN

O

N

O

O

O

OONH

O

N

O

N

NH

O

N

O

O

N

O

N

O

HN

288 / 1114

359 / 1043

987 / 289

1210 / 192

O

O

O

SOO

H2N

Ph


599 / 803

1113 / 289


33

NN

O

N

O

O

O

OONH

O

N

O

N

NH

O

N

O

O

N

O

N

O

HN

288 / 1119

359 / 1048

1006 / 401

1230 / 178

588 / 819O

OH

Ph

1088 / 319


Cl

Cl

OH


NN

O

N

O

O

O

OONH

O

N

O

N

NH

O

N

O

O

N

O

N

O

HN

288 / 849

359 / 778

849 / 383

989 / 241


430 / 802

912 / 320 NH

S

NH2

OO

HN

O

NHN

O


34

NN

O

N

O

O

O

OONH

O

N

O

N

NH

O

N

O

O

N

O

N

O

NH

288 / 849

359 / 778

928 / 209

999 / 138


922 / 319O

O

O

550 / 587858 / 279


NN

O

N

O

O

O

OONH

O

N

O

N

NH

O

N

O

O

N

O

N

O

NH

OH

288 / 953

359 / 882

428 / 811

848 / 393

1063 / 178F3C


O

OH

922 / 319


35

NN

O

N

O

O

O

OONH

O

N

O

N

NH

O

N

O

O

N

O

N

O

NH

OH

F


O

O

O

OH

288 / 1023

359 / 952

524 / 787

942 / 369

1133/ 178

992 / 319


NN

O

N

O

O

O

OONH

O

N

O

N

NH

O

N

O

O

N

O

N

O

NH

288 / 903

980 / 211

O

1093 / 97

909 / 282

N

O

359/ 832


O

SO O

H2N

949 / 344


36

NN

O

N

O

O

O

OONH

O

N

O

N

NH

O

N

O

O

N

O

N

O

NH

288 / 903

980 / 211

599 / 592


S O

OH2N

1093 / 97

909 / 282N

O

359/ 832


NN

O

N

O

O

O

OONH

O

N

O

N

NH

O

N

O

O

N

O

N

O

NH

288 / 1040

359 / 969

1027 / 301

599 / 729


Cl

Cl

S O

OH2N

OH

1256 / 72

329 / 997


37

NN

O

N

O

O

O

OONH

O

N

O

N

NH

O

N

O

O

N

O

N

O

NH

HO

S

288 / 809

359 / 738

512 / 585

848 / 249

919 / 178

860 / 237Exact Mass: 1095.5059


NN

O

N

O

O

O

OONH

O

N

O

N

NH

O

N

O

O

N

288 / 751NH

HN

O

O

O

N

O

HN

359 / 680

501 / 538

854 / 185

967 / 71 785

254



38

Hybrid DKP-capped library:

NN

NNH

O

O

O

O

O

NO

O

Exact Mass: 1018.53

O

O

N

N O

NO

O

N

N

O

O

O

474 / 546

614 / 406

729 / 292359 / 661518502


NN

NNH

O

O

O

O

O

NO

O

O

N O

N

N

O

O

O

N

N

CF3

OO

Exact Mass: 1112.48456 / 658

596 / 518

789 / 327

359 / 755

288 / 826484 / 630


39

NN

NNH

O

O

O

O

O

NO

O

O

N

N

O

O

N O

N

O

N

Exact Mass: 1128.50

NH

OHN

N

SNH2

O O

288 / 842

359 / 771

430 / 700

570 / 560

712 / 418

458 /

672


NN

NNH

O

O

O

O

O

NO

O

O

N

N

O

O

O

N

O

Exact Mass: 1267.51 O CF3

N

N

HN

CF3

288 / 981

359 / 910

590 / 679

730 / 539

827 / 442

616 / 653


40

NN

N

HN

O

O

O

O

O

N

OO O

N

N

O

O

N

O

N

HN

O

Exact Mass: 1058.52

N

H2N

O

S

288 / 772

359 / 702

456 / 604

596 / 464

749 / 312

482 / 576


NN

NNH

O

O

O

O

O

NO

O

O

O

N O

N

N

O

O


N

NHO

N

O

359 / 710

456 / 613

572

645 / 424

787 / 282

288/ 781


41

NN

N

HN

O

O

O

O

O

N

OO N O

N

N

O

OHN

O

S NH2O

O

OO

O N

Exact Mass: 1441.51

OF3C

OH

O

372 / 893

879 / 588

1117 / 348381 / 1083

288 / 1177


SNN

N

HN

O

O

O

O

O

NO

O

O

O

N

O

N

N

O

O

O


HN

N

N

O

O

HN O

381 / 781

524 / 637846 / 315

731 / 430


42

SNN

N

HN

O

O

O

O

O

NO

O

O

O

N

O

N

N

O

O

Exact Mass: 1258.49

N

O

O

O

O

HOS

NH2

O

O

310 / 993

381 / 921

496 / 809634 / 668

724 / 579

837 / 464

560 / 724


NN

NNH

O

O

O

O

O

NO

O

O

O

N

O

N

N

O

O

Exact Mass: 1171.50

N

N

OO

NH

O

S

359 / 814

288 / 885

550 / 623

578 / 595

704 / 469

857 / 316


43

NN

NNH

O

O

O

O

O

NO

O

O

N O

N

N

O

O

Exact Mass: 1180.49

S

O

N

N

OH

CF3

288 / 894

359 / 823

512 / 670

666 / 516

763 / 419

642

540


NN

N

HN

O

O

O

O

O

N

OO O

N

N

O

O

O

Exact Mass: 1041.56

N

N

O

N

HN

N

HN

O

472 / 571

626 / 417

723 / 320

500 / 543

359 / 684

288 / 755


44

NN

NNH

O

O

O

O

O

NO

O

O

N

N

O

O

N O

N

O

SNH2

OO

Exact Mass: 1316.50 (H+) 1338.50 (Na+)

N

O Ph

S

NH2

O

O

HN

N

310 / 1052

381 / 1003

452 / 910 682 / 680

922 / 441


NN

N

HN

O

O

O

O

O

N

OO

O

N

N

O

O

Exact Mass: 1230.48

O

N

NO

PhO

N

NH

N

686 / 546

CF3

S

456 / 546

839 / 393

359 / 874


45

NN

N

HN

O

O

O

O

O

N

OO

O

N

O

N

N

O

O

OO

SH2N

O

O

NH

Exact Mass: 1358.56

O

N

NO

Ph


NN

N

HN

O

O

O

O

O

N

OO

ON O

N

N

O

O

Exact Mass: 1218.52

F3C

O

O

N

NO

PhO

310 / 954

381 / 883

826 / 437

1017 / 225


46

ch cn and 0.1% trifluoroacetic acid (tfa) and buffer b (5m ...microwave assisted peptoid syntheses...

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