Skip to main content
. 2020 Nov 4;63(22):13709–13718. doi: 10.1021/acs.jmedchem.0c01202

Table 1. Phage Display Hits, Derivatives Thereof, and Their Prevention of α-Cbtx Inhibition of nAChRs.

peptide sequence fold prevention of α-Cbtx inhibitiona affinityb identified by
  no peptide (α-cbtx alone) 1.0    
1 Ac-HAKTYMWDGWYMPTSH-NH2 7.4 2 ± 2 μM phage displayc
2 Ac-HVNTYMWDGRYMRTSH-NH2 1.0   phage displayc
3 H-NWAPAQHGQKWYMQDYTSLE-NH2 3.8   phage displayd
4 H-TSNTTPWQTSWELMYAQQNY-NH2 1.6   phage displayd
5 H-KPHTYINDHNFIQYDNQQWL-NH2 not soluble   phage displayd
6 H-YVTHPWIDWWDNEPYM-NH2 not soluble   phage displayd
7 H-GHHHRVYMTDVTEHFRWMGK-NH2 0.6   phage displayd
8 H-GHHHRVYMTDVTEHFRWWVN-NH2 0.5   phage displayd
9 TAMRA-HAKTYMWDGWYMPTSH-NH2 not tested 52 ± 3 μM optimization
10 Ac-TYMWDGWYMPT-NH2 4.5   optimization
11 Ac-YMWDGWYM-NH2 6.9   optimization
12 [PEG]6-YMWDGWYM-NH2e 6.2 26 ± 4 μM optimization
13 [PEG]6-YMWDGW-OHe 0.3   optimization
14 [PEG]6-YMWDGW-NH2e 0.5   optimization
15 H-WDGWYM-[PEG]6e 0.3   optimization
16 Ac-WDGWYM-[PEG]6e 0.3   optimization
17 [PEG]6-YMWEGWYM-OHe 4.8   optimization
18 [PEG]6-YM(Nal)DGWYM-OHe,f 2.8   optimization
a

α-Cbtx (40 nM) inhibited ACh (100 μM) gated currents through nAChRs to 8 ± 2% their control level, which is here normalized to 1.0. In the presence of, e.g., peptide 1 (100 μM), α-Cbtx (40 nM) only inhibited ACh (100 μM) gated currents to 60 ± 18% of control, here normalized to 7.4-fold prevention of inhibition.

b

Kd value of 9 determined in fluorescence polarization (FP) saturation experiment (n = 2). Ki values for peptides 1 and 12 determined by inhibition of 9 binding to α-Cbtx in FP competition experiments (n = 6), only measured for two lead peptides.

c

After five rounds of selection by biopanning.

d

From deep sequencing of all phages.

e

PEG, polyethylene glycol.

f

Nal, 3-(2-naphthyl)-l-alanine.