Hydrogen exchange-mass spectrometry measures stapled peptide conformational dynamics and predicts pharmacokinetic properties.
Peptide drugs have traditionally suffered from poor pharmacokinetic properties due to their conformational flexibility and the interaction of proteases with backbone amide bonds. "Stapled Peptides" are cyclized using an all-hydrocarbon cross-linking strategy to reinforce their α-helical conformation, yielding improved protease resistance and drug-like properties. Here we demonstrate that hydrogen exchange-mass spectrometry (HX-MS) effectively probes the conformational dynamics of Stapled Peptides derived from the survivin-borealin protein-protein interface and predicts their susceptibility to proteolytic degradation. In Stapled Peptides, amide exchange was reduced by over five orders-of-magnitude versus the native peptide sequence depending on staple placement. Furthermore, deuteration kinetics correlated directly with rates of proteolysis to reveal the optimal staple placement for improved drug properties.
Aileron Therapeutics, Inc., 281 Albany Street, Cambridge, Massachusetts 02139, United States.
Xiangguo Eric
XE
Shi
Thomas E
TE
Wales
Carl
C
Elkin
Noriyuki
N
Kawahata
John R
JR
Engen
D Allen
DA
Annis
D000595:Amino Acid Sequence; D041961:Deuterium Exchange Measurement / Q000379:methods*; D005544:Forecasting; D006859:Hydrogen / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry* / Q000493:pharmacokinetics; D010599:Pharmacokinetics; D011487:Protein Conformation; D017434:Protein Structure, Tertiary
D010446:Peptide Fragments; D006859:Hydrogen