Table 1.
Sequence, helicity, cellular activity (EC50), and Mdm2 binding (Kd) of MP-292 and derivatives. Ratio refers to the fold-shift between the binding affinity and cellular activity. Section A) Alanine scan of MP-292 highlights F3, W7 and Cba10 as key residues for Mdm2 binding and cellular activity. Section B) D-amino acid scan of MP-292 shows that most variants maintain Mdm2 binding and cellular activity. Section C) D-amino acids substitutions to MP-189, a linear equivalent of stapled peptide MP-292, disrupt secondary structure and binding to Mdm2.
| Table section | Peptide | N-term | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | C-term | Helicity (%) | Cellular (µM) | Mdm2 (nM) | Ratio |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| A | MP-292 | Ac- | L | T | F | R8 | E | Y | W | Q | L | Cba | S5 | -SAA-amide | 32.8 | 0.54 | 18.6 | 29 |
| L1A | Ac- | A | T | F | R8 | E | Y | W | Q | L | Cba | S5 | -SAA-amide | 53.3 | 0.50 | 17.6 | 28 | |
| T2A | Ac- | L | A | F | R8 | E | Y | W | Q | L | Cba | S5 | -SAA-amide | 36.2 | 1.0 | 21.3 | 47 | |
| F3A | Ac- | L | T | A | R8 | E | Y | W | Q | L | Cba | S5 | -SAA-amide | 48.4 | 37 | 3600 | 10 | |
| E5A | Ac- | L | T | F | R8 | A | Y | W | Q | L | Cba | S5 | -SAA-amide | 27.6 | 0.18 | 29.8 | 6 | |
| Y6A | Ac- | L | T | F | R8 | E | A | W | Q | L | Cba | S5 | -SAA-amide | 50.3 | 0.55 | 38.9 | 14 | |
| W7A | Ac- | L | T | F | R8 | E | Y | A | Q | L | Cba | S5 | -SAA-amide | ND | 8.8 | 5093 | 2 | |
| Q8A | Ac- | L | T | F | R8 | E | Y | W | A | L | Cba | S5 | -SAA-amide | 33.4 | 0.10 | 26.5 | 4 | |
| L9A | Ac- | L | T | F | R8 | E | Y | W | Q | A | Cba | S5 | -SAA-amide | 48.7 | 0.16 | 12.0 | 13 | |
| Cba10A | Ac- | L | T | F | R8 | E | Y | W | Q | L | A | S5 | -SAA-amide | ND | 11 | 176 | 61 | |
| S12A | Ac- | L | T | F | R8 | E | Y | W | Q | L | Cba | S5 | -AAA-amide | 36.5 | 0.35 | 20.4 | 17 | |
| B | L1(D-Leu) | Ac- | D-Leu | T | F | R8 | E | Y | W | Q | L | Cba | S5 | -SAA-amide | 44.1 | 0.6 | 31 | 18 |
| T2(D-Thr) | Ac- | L | D-Thr | F | R8 | E | Y | W | Q | L | Cba | S5 | -SAA-amide | 40.7 | 3.4 | 136 | 25 | |
| F3(D-Phe) | Ac- | L | T | D-Phe | R8 | E | Y | W | Q | L | Cba | S5 | -SAA-amide | 51.8 | >50 | 6485 | >8 | |
| E5(D-Glu) | Ac- | L | T | F | R8 | D-Glu | Y | W | Q | L | Cba | S5 | -SAA-amide | 38.2 | 1.0 | 32 | 30 | |
| Y6 (D-Tyr) | Ac- | L | T | F | R8 | E | D-Tyr | W | Q | L | Cba | S5 | -SAA-amide | 43.2 | 3.2 | 286 | 11 | |
| W6(D-Trp) (MP-384) | Ac- | L | T | F | R8 | E | Y | D-Trp | Q | L | Cba | S5 | -SAA-amide | 41.0 | 0.6 | 30 | 20 | |
| Q8(D-Gln) | Ac- | L | T | F | R8 | E | Y | W | D-Gln | L | Cba | S5 | -SAA-amide | 27.1 | 2.1 | 22 | 95 | |
| L9(D-Leu) | Ac- | L | T | F | R8 | E | Y | W | Q | D-Leu | Cba | S5 | -SAA-amide | 39.0 | 1.1 | 30 | 37 | |
| Cba10(D-Cba) | Ac- | L | T | F | R8 | E | Y | W | Q | L | D-Cba | S5 | -SAA-amide | ND | 8.1 | 1650 | 5 | |
| S12(D-Ser) | Ac- | L | T | F | R8 | E | Y | W | Q | L | Cba | S5 | -(D-Ser)-AA-amide | 41.2 | 1.2 | 8.6 | 140 | |
| A13(D-Ala) | Ac- | L | T | F | R8 | E | Y | W | Q | L | Cba | S5 | -S-(D-Ala)-A-amide | 41.8 | 0.6 | 17 | 35 | |
| A14(D-Ala) | Ac- | L | T | F | R8 | E | Y | W | Q | L | Cba | S5 | -SA-(D-Ala)-amide | 34.6 | 0.5 | 18 | 28 | |
| C | Linear Parent (MP-189) | Ac- | L | T | F | Aib | E | Y | W | Q | L | Cba | Aib | -SAA-amide | 35.3 | 33.5 | 43.1 | 777 |
| Linear L1(D-Leu) | Ac- | D-Leu | T | F | Aib | E | Y | W | Q | L | Cba | Aib | -SAA-amide | 37.2 | >50 | 277 | >180 | |
| Linear E5(D-Glu) | Ac- | L | T | F | Aib | D-Glu | Y | W | Q | L | Cba | Aib | -SAA-amide | 21.4 | >50 | 135 | >370 | |
| Linear Q8(D-Gln) | Ac- | L | T | F | Aib | E | Y | W | D-Gln | L | Cba | Aib | -SAA-amide | 21.8 | >50 | 119 | >370 | |
| Linear L9(D-Leu) | Ac- | L | T | F | Aib | E | Y | W | Q | D-Leu | Cba | Aib | -SAA-amide | 19.7 | >50 | 88 | >568 |