Bradykinin analogues with ?-amino acid substitutions reveal subtle differences in substrate specificity between the endopeptidases EC 3.4.24.15 and EC 3.4.24.16

DPP‐4 Cleaves α/β‐Peptide Bonds: Substrate Specificity and Half‐Lives

“…[9,10] Besides pure β-peptides, it has been shown that α/βpeptides in which the β-amino acid replacement does not change the relative orientation of side chains around the scissile bond (Scheme 1), thus mimicking α-peptide environment, are still stable against a wide range of proteases. [9] This was confirmed by many examples of peptide analogs where a strategic replacement of a single P1' amino acid by a βhomolog (Scheme 1b) abolished the activity of pepsin, chymotrypsin, plasmepsin II, [11] metalloproteinase EP24.15 and EP24.16, [12] neprylisin, [13] pronase E [14] or ACE-2, [15] and improved the stability of the whole peptide. With this kind of substitution, minimal modifications are possible in order to provide peptide stability, while maintaining α-peptide structural and spatial characteristics required for interaction with receptor.…”

DPP‐4 Cleaves α/β‐Peptide Bonds: Substrate Specificity and Half‐Lives

“…[9,10] Besides pure β-peptides, it has been shown that α/βpeptides in which the β-amino acid replacement does not change the relative orientation of side chains around the scissile bond (Scheme 1), thus mimicking α-peptide environment, are still stable against a wide range of proteases. [9] This was confirmed by many examples of peptide analogs where a strategic replacement of a single P1' amino acid by a βhomolog (Scheme 1b) abolished the activity of pepsin, chymotrypsin, plasmepsin II, [11] metalloproteinase EP24.15 and EP24.16, [12] neprylisin, [13] pronase E [14] or ACE-2, [15] and improved the stability of the whole peptide. With this kind of substitution, minimal modifications are possible in order to provide peptide stability, while maintaining α-peptide structural and spatial characteristics required for interaction with receptor.…”

Thimet Oligopeptidase Biochemical and Biological Significances: Past, Present, and Future Directions

“…This cleavage by NEP generates a potent inhibitor of angiotensin converting enzyme 1 (ACE1), thereby limiting the use of CFP for in vivo studies. Additional inhibitors of THOP1 based on the CFP were developed such as N-[1-(R, S)-carboxy-3-phenylpropyl]-A-Aib-Y-p-aminobenzoate (JA2), which was resistant to in vivo proteolysis after incorporation of beta-amino acids [117,127,160]. JA2 is a mixed inhibitor of THOP1 and the structurally homologous enzyme neurolysin (EC3.4.24.16; Nln), which was resistant to cleavage by the related metalloendopeptidase NEP, in contrast to the precursor CFP inhibitor [117,127,160].…”

“…Additional inhibitors of THOP1 based on the CFP were developed such as N-[1-(R, S)-carboxy-3-phenylpropyl]-A-Aib-Y-p-aminobenzoate (JA2), which was resistant to in vivo proteolysis after incorporation of beta-amino acids [117,127,160]. JA2 is a mixed inhibitor of THOP1 and the structurally homologous enzyme neurolysin (EC3.4.24.16; Nln), which was resistant to cleavage by the related metalloendopeptidase NEP, in contrast to the precursor CFP inhibitor [117,127,160]. The potent pharmacological in vivo effects of JA2 was seen in the hypotension induced by bradykinin, which did not diminish even 4 h after JA2 injection [127].…”

Inhibitors of Metalloendopeptidase EC 3.4.24.15 and EC 3.4.24.16 Stabilized against Proteolysis by the Incorporation of β-Amino Acids

“…β-Amino acids have recently reemerged as a potential peptidomimetic tool in the design of metabolically stable bioactive peptides (28)(29)(30)(31)(32)(33)(34). We recently reported the synthesis of a selection of CFP analogues incorporating β-amino acids which retained inhibitory activity, demonstrating that β-amino acid-containing peptides are able to bind to target proteolytic enzymes (30).…”

DPP‐4 Cleaves α/β‐Peptide Bonds: Substrate Specificity and Half‐Lives

“…[9,10] Besides pure β-peptides, it has been shown that α/βpeptides in which the β-amino acid replacement does not change the relative orientation of side chains around the scissile bond (Scheme 1), thus mimicking α-peptide environment, are still stable against a wide range of proteases. [9] This was confirmed by many examples of peptide analogs where a strategic replacement of a single P1' amino acid by a βhomolog (Scheme 1b) abolished the activity of pepsin, chymotrypsin, plasmepsin II, [11] metalloproteinase EP24.15 and EP24.16, [12] neprylisin, [13] pronase E [14] or ACE-2, [15] and improved the stability of the whole peptide. With this kind of substitution, minimal modifications are possible in order to provide peptide stability, while maintaining α-peptide structural and spatial characteristics required for interaction with receptor.…”

Thimet Oligopeptidase Biochemical and Biological Significances: Past, Present, and Future Directions

“…This cleavage by NEP generates a potent inhibitor of angiotensin converting enzyme 1 (ACE1), thereby limiting the use of CFP for in vivo studies. Additional inhibitors of THOP1 based on the CFP were developed such as N-[1-(R, S)-carboxy-3-phenylpropyl]-A-Aib-Y-p-aminobenzoate (JA2), which was resistant to in vivo proteolysis after incorporation of beta-amino acids [117,127,160]. JA2 is a mixed inhibitor of THOP1 and the structurally homologous enzyme neurolysin (EC3.4.24.16; Nln), which was resistant to cleavage by the related metalloendopeptidase NEP, in contrast to the precursor CFP inhibitor [117,127,160].…”

“…Additional inhibitors of THOP1 based on the CFP were developed such as N-[1-(R, S)-carboxy-3-phenylpropyl]-A-Aib-Y-p-aminobenzoate (JA2), which was resistant to in vivo proteolysis after incorporation of beta-amino acids [117,127,160]. JA2 is a mixed inhibitor of THOP1 and the structurally homologous enzyme neurolysin (EC3.4.24.16; Nln), which was resistant to cleavage by the related metalloendopeptidase NEP, in contrast to the precursor CFP inhibitor [117,127,160]. The potent pharmacological in vivo effects of JA2 was seen in the hypotension induced by bradykinin, which did not diminish even 4 h after JA2 injection [127].…”

Inhibitors of Metalloendopeptidase EC 3.4.24.15 and EC 3.4.24.16 Stabilized against Proteolysis by the Incorporation of β-Amino Acids

“…β-Amino acids have recently reemerged as a potential peptidomimetic tool in the design of metabolically stable bioactive peptides (28)(29)(30)(31)(32)(33)(34). We recently reported the synthesis of a selection of CFP analogues incorporating β-amino acids which retained inhibitory activity, demonstrating that β-amino acid-containing peptides are able to bind to target proteolytic enzymes (30).…”

DPP‐4 Cleaves α/β‐Peptide Bonds: Substrate Specificity and Half‐Lives

“…[9,10] Besides pure β-peptides, it has been shown that α/βpeptides in which the β-amino acid replacement does not change the relative orientation of side chains around the scissile bond (Scheme 1), thus mimicking α-peptide environment, are still stable against a wide range of proteases. [9] This was confirmed by many examples of peptide analogs where a strategic replacement of a single P1' amino acid by a βhomolog (Scheme 1b) abolished the activity of pepsin, chymotrypsin, plasmepsin II, [11] metalloproteinase EP24.15 and EP24.16, [12] neprylisin, [13] pronase E [14] or ACE-2, [15] and improved the stability of the whole peptide. With this kind of substitution, minimal modifications are possible in order to provide peptide stability, while maintaining α-peptide structural and spatial characteristics required for interaction with receptor.…”

Thimet Oligopeptidase Biochemical and Biological Significances: Past, Present, and Future Directions

“…This cleavage by NEP generates a potent inhibitor of angiotensin converting enzyme 1 (ACE1), thereby limiting the use of CFP for in vivo studies. Additional inhibitors of THOP1 based on the CFP were developed such as N-[1-(R, S)-carboxy-3-phenylpropyl]-A-Aib-Y-p-aminobenzoate (JA2), which was resistant to in vivo proteolysis after incorporation of beta-amino acids [117,127,160]. JA2 is a mixed inhibitor of THOP1 and the structurally homologous enzyme neurolysin (EC3.4.24.16; Nln), which was resistant to cleavage by the related metalloendopeptidase NEP, in contrast to the precursor CFP inhibitor [117,127,160].…”

“…Additional inhibitors of THOP1 based on the CFP were developed such as N-[1-(R, S)-carboxy-3-phenylpropyl]-A-Aib-Y-p-aminobenzoate (JA2), which was resistant to in vivo proteolysis after incorporation of beta-amino acids [117,127,160]. JA2 is a mixed inhibitor of THOP1 and the structurally homologous enzyme neurolysin (EC3.4.24.16; Nln), which was resistant to cleavage by the related metalloendopeptidase NEP, in contrast to the precursor CFP inhibitor [117,127,160]. The potent pharmacological in vivo effects of JA2 was seen in the hypotension induced by bradykinin, which did not diminish even 4 h after JA2 injection [127].…”

Inhibitors of Metalloendopeptidase EC 3.4.24.15 and EC 3.4.24.16 Stabilized against Proteolysis by the Incorporation of β-Amino Acids

“…β-Amino acids have recently reemerged as a potential peptidomimetic tool in the design of metabolically stable bioactive peptides (28)(29)(30)(31)(32)(33)(34). We recently reported the synthesis of a selection of CFP analogues incorporating β-amino acids which retained inhibitory activity, demonstrating that β-amino acid-containing peptides are able to bind to target proteolytic enzymes (30).…”