Table 2.
Sequence, helicity, cellular activity (EC50), and Mdm2 binding (Kd) of MP-081 and derivatives. A variety of putative helix inducers and helix-breakers are accommodated in the context of stapled peptide MP-081 with most variants maintaining helical structure, Mdm2 binding and cellular activity. Ratio refers to the fold-shift between the binding affinity and cellular activity.
| Peptide | N-term | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | C-term | Helicity (%) | Cellular (µM) | Mdm2 (nM) | Ratio |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| MP-081 | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -SAA-amide | 49.6 | 0.42 | 19.0 | 22 |
| L1(Aib) | Ac-K(N3)-(βA)- | Aib | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -SAA-amide | 42.7 | 0.49 | 29.5 | 17 |
| T2(Aib) | Ac-K(N3)-(βA)- | L | Aib | F | R8 | E | Y | W | A | Q | Cba | S5 | -SAA-amide | 38.3 | 2.9 | 1706 | 2 |
| A8(Aib) | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | Aib | Q | Cba | S5 | -SAA-amide | 37.5 | 1.5 | 119.4 | 12 |
| Q9(Aib) | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | A | Aib | Cba | S5 | -SAA-amide | 28 | 0.25 | 33.1 | 8 |
| S12(Aib) | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -(Aib)-AA-amide | 37.3 | 0.25 | 24.4 | 10 |
| A13(Aib) | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -A-(Aib)-A-amide | 34.4 | 0.59 | 25.7 | 23 |
| L1(N-methyl L-Leu) | Ac-K(N3)-(βA)- | NMe-L | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -SAA-amide | 32.6 | 0.46 | 23.5 | 20 |
| L1(N-methyl L-Ala) | Ac-K(N3)-(βA)- | NMe-A | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -SAA-amide | 47.2 | 1.49 | 15.4 | 97 |
| T2(N-methyl L-Thr) | Ac-K(N3)-(βA)- | L | NMe-T | F | R8 | E | Y | W | A | Q | Cba | S5 | -SAA-amide | 38.2 | 7.7 | 352.2 | 22 |
| A8(N-methyl L-Ala) | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | NMe-A | Q | Cba | S5 | -SAA-amide | 16.9 | >50 | 671.3 | >75 |
| Q9(N-methyl L-Gln) | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | A | NMe-Q | Cba | S5 | -SAA-amide | 20.2 | >50 | 211.2 | >237 |
| A13(N-methyl L-Ala) | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -S-(Nme-A)-A-amide | 38.6 | 1.6 | 10.0 | 158 |
| A14(N-methyl L-Ala) | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -SA-(Nme-A)-amide | 47.5 | 0.71 | 5.9 | 120 |
| L1P | Ac-K(N3)-(βA)- | P | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -SAA-amide | 41.6 | 0.75 | 21.9 | 34 |
| Q9P | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | A | P | Cba | S5 | -SAA-amide | 27.4 | 3.1 | 77.1 | 41 |
| S12P | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -PAA-amide | 30.8 | 3.7 | 45.7 | 80 |
| L1G | Ac-K(N3)-(βA)- | G | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -SAA-amide | 44.6 | 1.6 | 22.1 | 73 |
| E5G | Ac-K(N3)-(βA)- | L | T | F | R8 | G | Y | W | A | Q | Cba | S5 | -SAA-amide | 28.6 | 0.26 | 61.8 | 4 |
| Q9G | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | A | G | Cba | S5 | -SAA-amide | 38.8 | 0.68 | 42.0 | 16 |
| S12G | Ac-K(N3)-(βA)- | L | T | F | R8 | E | Y | W | A | Q | Cba | S5 | -GAA-amide | 46.5 | 0.65 | 18.9 | 34 |
| L1G, E5G, A8G | Ac-K(N3)-(βA)- | G | T | F | R8 | G | Y | W | G | Q | Cba | S5 | -SAA-amide | 20.4 | 1.7 | 49.8 | 34 |