Metal Species forAmide Hydrolysis

Literature Seminar, 20130511
Kiyomichi SHINODA (M1)1

Table of Contents§1 Introduction§2 Residue- or Sequence-Selective Hydrolysis of Amides§3 Protein-Selective Hydrolysis of Amides2§1 Introduction3

§1 Introduction

Characteristics of Amide Bond
● stability toward hydrolytic cleavage
O
Me

N
H

H
N

O

O

t1/2 = 500 years
OH

O

pH = 6.8, 25 °C

Me

N
H

OH +
H2N
O

O
OH

A. Radzicka, R. Wolfenden, J. Am. Chem. Soc. 1996, 118, 6105-6109.

● This stability derives from resonance effect.
resonance
effect

O
R1

N
H

R2

inactive toward
nucleophilic attack

OH
R1

N

R2

4

§1 Introduction

Characteristics of Amide Bond
● planar structure due to the resonance effectresonance
effect

O
R1

N
H

R2

R1

OH
R1

N
N

R2

H

O
R2

○ twist angle distribution of
tertiary amides(A. J. Kirby et al., J. Chem. Soc., Perkin Trans. 2 2001, 522.)5§1 Introduction

Destabilizing Amide Bond
● activation by Lewis acidLA
O
R1

Lewis acid
R2

N
H

δ+

O

R1

N
H

R2

O

cleavage
R1+ H2N R2OH

● activation by bond distortion
R1

O

bond
distortion

O

N

R2

N

R2
H

O

R2

N
H
R1
6

H

N
R1

H

≡

≡
N

R2

R1

H

O
R1

cleavage

δ+

H

O
+
HO

R2

§1 Introduction

Precedents: Activation by Lewis Acid
● hydrolysis by cerium(IV)-cyclodextrin complex
O

H
N

H2N

Ce(NH4)2(NO3)6 (1 eq.)γ-CD (5 eq.)
OH

O

Ph

O
OH

H2N

0.1 M Tris buffer (0.01 M)
pH 8.0, 60 °C, 24 h

H2N

+

OH

O
Ph
39% conversion

M. Komiyama et al., J. Chem. Soc., Chem. Commun. 1994, 1758.

● Zinc-catalyzed solvolysis

Cbz

H
N

OH

O
N
H

Zn(OTf)2 (5 mol%)diethylcarbonate (2 eq.)

OMeO

n-BuOH (2 M)
reflux, 45 h

Cbz

H
N

O
On-Bu

84% yield

K. Mashima et al., Angew. Chem. Int. Ed. 2012, 51, 5723.
7

§1 Introduction

Precedents: Activation by Bond Twisting● molecules containing twisted amide
NO

total synthesis of○ the former: K. Tani & B. M. Stoltz,
Nature 2006, 441, 731.
(tetrafluoroborate salt)○ the latter: A. J. Kirby et al., J. Chem. Soc.,
Perkin Trans. 2, 2001, 522.

NO2-quinoclidone

1-aza-2-adamantanone(Kirby's Amide)○ characters

N
N

O

BF4

O
N
H
O

twist angle

2.5°

90.5°

90.9°

bond lengthC-N

1.325 Å1.475 Å1.526 Å8

(Values are cited from the above 2 papers .)§1 Introduction

Precedents: Activation by Bond Twisting● hydrolysis of twisted amides
CO2D
BF4
N
H

t1/2 < 15 s
D2O, r.t.
N
H

O
N

O

NH2
COO

H2O
over a few minutes

K. Tani & B. M. Stoltz, Nature 2006, 441, 731.
A. J. Kirby et al., J. Am. Chem. Soc., 1998, 120, 7101.

9§1 Introduction

In This Seminar...
● activation by Lewis acidLA
O
R1

Lewis acid
R2

N
H

δ+

O

R1

N
H

R2

O

cleavage
R1+ H2N R2OH

● activation by bond distortion
R1

O

bond
distortion

O

N

R2

N

R2
H

O

R2

N
H
R1
10

H

N
R1

H

≡

≡
N

R2

R1

H

O
R1

cleavage

δ+

H

O
+
HO

R2

§2 Residue- or Sequence-Selective Hydrolysis of Amides

11§2 Residue- or Sequence-Selective Hydrolysis of Amides

Palladium(II) Complexes
H2O
H2O

Pd

H2O OH2
Pd
H2N
NH2

OH2
OH2

[Pd(H2O)4]2+

H2O OH2
β-CD
Pd
MeS
S

[Pd(en)(H2O)2]2+
OH
O

HO

O

β-CD =

OHOHO

O
OH
O

HO
O
OH

OH

HO

O
OH
O
OH

OH
O
OH

OH

O

OH
OH O

OH
OOH

HO

O
OH

12

O

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(H2O)4]2+● substrate: Met-peptide
Ac-Ala-Lys-Tyr-Gly-Gly-Met-Ala-Ala-Arg-Ala-OH● hydrolysis condition
Met-peptide[Pd(H2O)4]2+ (1 eq.)fragments

aqueous media (1 mM)
pH 2.3, 60 °C
24 h

N. M. Kostić et al., J. Am. Chem. Soc. 2002, 124, 4759.
13

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(H2O)4]2+● experiment procedure
60 °C
5 min

color change

1h

12 h

22 h

24 h

HPLC sample HPLC sample HPLC sample○ 5 min: [Pd(H2O)4]2+ was bound to the peptide.
(checked by HPLC and MALDI MS)○ 24 h: two components were observed.
N. M. Kostić et al., J. Am. Chem. Soc. 2002, 124, 4759.
14§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(H2O)4]2+● HPLC charts

60 °C
5 min

1h

color change

12 h

22 h

24 h

HPLC sample HPLC sample HPLC sample

2 peaks were separated by HPLC
and MALDI mass spectroscopic
identification was conducted.

N. M. Kostić et al., J. Am. Chem. Soc. 2002, 124, 4759.
15§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(H2O)4]2+● composition of the fractions

H-Gly-Met-Ala-Ala-Arg-Ala-OH
Ac-Ala-Lys-Tyr-Gly-OH!
Ac-Ala-Lys-Tyr-Gly-Gly-Met-Ala-Ala-Arg-Ala-OH
N. M. Kostić et al., J. Am. Chem. Soc. 2002, 124, 4759.
16

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(H2O)4]2+● TOCSY 1H NMR spectrum

before additionof Pd complex

5 min after
the addition

N. M. Kostić et al., J. Am. Chem. Soc. 2002, 124, 4759.

○ SMe singlet: 2.12 ppm → 2.45 ppm＝ Pd(II) complex was bound to the Met6 side chain.
17§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(H2O)4]2+● TOCSY 1H NMR spectrum

before additionof Pd complex

5 min after
the addition

N. M. Kostić et al., J. Am. Chem. Soc. 2002, 124, 4759.

○ Met6NH-CH signal disappeared: deprotonated＝ complete coordination of amide to the Pd(II) ion
18§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(H2O)4]2+● TOCSY 1H NMR spectrum

24 h after
the addition(reaction finished)

before additionof Pd complex

N. M. Kostić et al., J. Am. Chem. Soc.
2002, 124, 4759.

○ Signals derived from Gly5 and Met6 disappeared.
＝ Amide cleavage occurred at Gly4-Gly5.
19

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(H2O)4]2+● equilibrium among Pd(II) complexes

activeinactive○ 2 and 3 exists at pH 2.3,
and 3 is major.

N. M. Kostić et al., J. Am. Chem. Soc. 2002, 124, 4759.
20§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(H2O)4]2+● 2 is hydrolytically active, but 3 is hydrolytically inactive.
O

O
N

N
H

HN
Pd

O

S

H
N
O

OH
O

hydrolysis

NH

OH2
NH

O

O
N

N
O

Pd

N
H
S

O

HN Pd

O

H
N

N
H

O

S

OH2

H
N

O
OH

hydrolysis

O

OH2
NH

N

+

O

2

O

O

O

O
N

+

N

Pd

N
H
S

H
N
O

O

3

NH2

coordination of 2 amide nitrogen atoms→ Lewis acidity of Pd(II) ion was quenched.
21

N. M. Kostić et al., J. Am. Chem. Soc.
2002, 124, 4759.

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(en)(H2O)2]2+● substrate: oxidized chain B of bovine insulin
H-Phe-Val-Asn-Gln-His-Leu-CysOX-Gly-Ser-His-Leu-Val-Glu-Ala-Leu-Tyr-Leu-Val-CysOX-Gly-Glu-Arg-Gly-Phe-Phe-Tyr-Thr-Pro-Lys-Ala-OH● hydrolysis condition
oxidized chain Bof bovine insulin[Pd(en)(H2O)2]2+ (5 eq.)fragments

aqueous media (1 mM)
pH 2.0, 60 °C
4 daysN. M. Kostić et al., Inorg. Chem. 2002, 41, 7053.
22

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(en)(H2O)2]2+● cleavage site!

!

H-Phe-Val-Asn-Gln-His-Leu-CysOX-Gly-Ser-His-Leu-Val-Glu-Ala-Leu-Tyr-Leu-Val-CysOX-Gly-Glu-Arg-Gly-Phe-Phe-Tyr-Thr-Pro-Lys-Ala-OH○ Pd(II) reagent promoted cleavage of the second bond
upstream from the His5 and His10 anchor.
(characterized by MALDI mass spectroscopic identification
and amino acid analysis: N-terminal Edman degradation)
N. M. Kostić et al., Inorg. Chem. 2002, 41, 7053.
23

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Acidic Hydrolysis with [Pd(en)(H2O)2]2+● equilibrium among Pd(II) complexes
R1
N
H

O

H
N
O

R1

O
N
H
OH2

R2

N
H

O

H
N

N
R2

O

H2N PdH2N Pd N

N

NH2

NH

NH2

O

NH
[Pd(en)(H2O)2]2+

1○ 2 and 3 exists at pH 2.0[Pd(en)2]2+ + 3H3O+
O

O

O
H
N
R1
HN

O

N

R2
O
H2O Pd
OH2

R2

NH

O

N
N Pd

O

N

O

R2

N

OH2

NH

HN

3

active

24

N Pd

O

R1

2

N

inactive

N

N
NH

R1

4

N. M. Kostić et al., Inorg. Chem. 2002, 41, 7053.

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Neutral Hydrolysis with [Pd(H2O)4]2+!

● substrates & cleavage site

GlyMet-peptide: Ac-Ala-Lys-Tyr-Gly-Gly-Met-Ala-Ala-Arg-Ala-OH!

SarMet-peptide: Ac-Lys-Gly-Gly-Ala-Gly-Sar-Met-Ala-Ala-Arg-Gly-OH!

ProMet-peptide: Ac-Lys-Gly-Gly-Ala-Gly-Pro-Met-Ala-Ala-Arg-Gly-OH

25

N. M. Kostić et al., J. Am. Chem. Soc. 2003, 125, 781.

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Neutral Hydrolysis with [Pd(H2O)4]2+● dependence on pH of the 1st-order-rate const.
○ reaction:

peptide[Pd(H2O)4]2+ (1 eq.)fragments

aqueous media (1 mM)
60 °C

Why?
ProMet > SarMet > GlyMet

ProMet- and SarMet-peptide can be cleaved but GlyMet-peptide can’t.

Why?
26

N. M. Kostić et al., J. Am. Chem. Soc. 2003, 125, 781.

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Neutral Hydrolysis with [Pd(H2O)4]2+● difference between GlyMet- (left) and ProMet- (right)
peptide in equilibrium among Pd(II) complexes
R
N
H

O

H
N

O
N
H
OH2

O

H2O Pd S

O

O
H
N
O

H2O PdHN

active (minor*)

1

S

1Pro

N Pd

S

O

N

OH2
R

R

N

NH

O

S

4

inactive (major*)

activeH2O Pd S
OH2

3
27

O
N

HN

R

(* at pH 2.3)

O

N

N

O

OH2

O
O

N Pd

H2O Pd S

2

O
O

N
H
OH2

O

OH2

OH2

O

O
N

N
H

N

R

R

2Pro

N. M. Kostić et al., J. Am. Chem. Soc. 2003, 125, 781.

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Neutral Hydrolysis with [Pd(H2O)4]2+● TOCSY 1H NMR spectrum (ProMet-peptide)
before additionof Pd complex○ signals derived from
Met7NH disappeared.
↓
Met7NH group
coordinates to Pd(II) ion

10 min after
addition of 1 eq.
Pd complex○ NH signals of otherresidues persisted.
↓
Other amide groups were
not bound either.

28

N. M. Kostić et al., J. Am. Chem. Soc. 2003, 125, 781.

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Neutral Hydrolysis with [Pd(H2O)4]2+● external attack vs internal delivery○ in this case...

29

N. M. Kostić et al., J. Am. Chem. Soc. 2003, 125, 781.

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Neutral Hydrolysis with [Pd(H2O)4]2+● external attack vs internal delivery
O
H

O
H

N

N

O

O

H

H2O Pd S
HN

H

OH2

O
NH

cis-Gly-Pro

N

N

ROESY

O

Pd S
OH2

trans-Gly-Pro○ ROESY 1H NMR spectrum showed the cross peak
between the Gly5α-CH and Pro6δ-CH→ external attack (trans-Gly-Pro)
30

N. M. Kostić et al., J. Am. Chem. Soc. 2003, 125, 781.

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Neutral Hydrolysis with Pd(II)-β-CD complex● substrate: Met-peptide
Ac-Lys-Gly-Gly-Phe-Ser-Pro-Phe-Phe-Ala-Ala-Arg-Ala-OH● hydrolysis condition
H2O OH2β-CD
Pd(10 eq.)
MeS
S
peptidefragments
aqueous media (1 mM)
pH 7.5, 60 °C
24 h

31

N. M. Kostić et al., J. Am. Chem. Soc. 2004, 126, 696.

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Neutral Hydrolysis with Pd(II)-β-CD complex● cleavage site!

Ac-Lys-Gly-Gly-Phe-Ser-Pro-Phe-Ala-Ala-Arg-Ala-OH○ Pd(II)-β-CD complex promoted cleavage of thefirst one upstream from the Pro6-Phe7 sequence.
(characterized by MALDI mass spectroscopic identification)

32

N. M. Kostić et al., J. Am. Chem. Soc. 2004, 126, 696.

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Neutral Hydrolysis with Pd(II)-β-CD complex● possible interaction toward sequence selectivity○ CD cavities bind hydrophobic substrates in aqueous media.
33

N. M. Kostić et al., J. Am. Chem. Soc. 2004, 126, 696.

§2 Residue- or Sequence-Selective Hydrolysis of Amides

Summary of This Section● residue-selectivehydrolysis● sequence-selectivehydrolysis

N. M. Kostić et al., J. Am. Chem. Soc. 2003, 125, 781.
N. M. Kostić et al., J. Am. Chem. Soc. 2004, 126, 696.
34

§3 Protein-Selective Hydrolysis of Amides35

§3 Protein-Selective Hydrolysis of Amides

Mb-Selective Artificial Protease● catalyst design○ catalyst site
: Cyc-metal complex○ binding site
: peptide nucleic acid (PNA)

J. Suh et al., Bioorg. Med. Chem. 2003, 11, 2901.
36

§3 Protein-Selective Hydrolysis of Amides

Mb-Selective Artificial Protease● Based on this design, the library of Cyc-containing
PNA oligomers was constructed.
○ n=7, 8: no activities* toward Mb (Cu(II)-complex)○ n=9: active* toward Mb (Cu(II)-complex)(* checked by SDS-PAGE)

J. Suh et al., Bioorg. Med. Chem. 2003, 11, 2901.
37

§3 Protein-Selective Hydrolysis of Amides

Mb-Selective Artificial Protease● their own combinatorial library○ Cu(II)1 was the best.
J. Suh et al., Bioorg. Med. Chem. 2003, 11, 2901.
38

§3 Protein-Selective Hydrolysis of Amides

Mb-Selective Artificial Protease● changing metal center from Cu(II) to Co(III)○ Metal transfer to metal-abstracting materials in living bodyshould be slower for Co(III) complexes because of theexchange-inertness of Co(III)

Cu(II) 2.0 μM○ Furthermore, Co(III)1 showshigh reactivity than Cu(II)1.
k0(Cu) = 5.3☓10-3 h-1
k0(Co) = 9.4☓10-3 h-1

Co(III) 0.47 μM(pseudo-1st-order kinetics)
decrease of [Mb]
(pH 7.5, 37 ℃)

J. Suh et al., Bioorg. Med. Chem. 2003, 11, 2901.
39

§3 Protein-Selective Hydrolysis of Amides

Mb-Selective Artificial Protease● affinity of Co(III)1 complex to Mb○ Two straight lines intersect
at C0 = [Mb]0 = 4.7 μM.
↓[Mb]0 = 4.7 μM
pH= 7.5, 37 ℃

○ Km < 4.7 μM < 5.0 μM(cf. Km of fumarase = 5.0 μM)[Mb}0→ Affinity of Co(III)1 complex to Mb was high enough.
→ kcat (not kcat/Km) should be considered!!

J. Suh et al., Bioorg. Med. Chem. 2003, 11, 2901.

40

§3 Protein-Selective Hydrolysis of Amides

Mb-Selective Artificial Protease● kcat comparison between Co(III) complexes

Co(III)1 was the best!

● pH dependence of kcat for cleavage of
Mb by Co(III)1 complex→ most active at physiological pH!
J. Suh et al., Bioorg. Med. Chem. 2003, 11, 2901.

41

§3 Protein-Selective Hydrolysis of Amides

Mb-Selective Artificial Protease● possible cleavage mechanism○ This mechanism consistents
with the pH dependence ofCo(III)1 complex.
||○ The following two would
reduce the reactivity ofCo(III)1 complex.
✓ protonation of the hydroxo (T)or oxo (T’) ligand at low pH✓ deprotonation of the ammonium
ion (T’) at high pH
42

r.d.s.

J. Suh et al., J. Biol. Inorg. Chem. 2009, 14, 151.

§3 Protein-Selective Hydrolysis of Amides

Mb-Selective Artificial Protease● cleavage site characterization○ N-terminal sequencing (Edman degradation) was
conducted to product solution of Co(III)3.
→ Leu89-Ala90 was the cleavage site.
J. Suh et al., Bioorg. Med. Chem. 2003, 11, 2901.
43

§3 Protein-Selective Hydrolysis of Amides

Mb-Selective Artificial Protease● selectivity (control experiment)
albumin, γ-globulin, elongation factor P,
gelatin A, gelatin B○ Above 5 proteins were incubated with Cu(II)1 or Co(III)1,
but protein cleavage was not observed.

Target selectivity can be expected!
J. Suh et al., Bioorg. Med. Chem. 2003, 11, 2901.
44

§3 Protein-Selective Hydrolysis of Amides

PDF-Selective Artificial Protease● PDF: peptide deformylase○ involved in deformylation of the formyl-methionyl
derivative of proteins formed in the prokaryotic
translational systems.

○ Its inhibitors are searched as candidates for new
antibiotic drugs.

way toward “catalytic drugs”
J. Suh et al., J. Am. Chem. Soc. 2005, 127, 2396.
45§3 Protein-Selective Hydrolysis of Amides

PDF-Selective Artificial Protease● construction of a library of catalyst candidates○ synthesized by the Ugi reaction (product was racemic mixture)
O
+
R1

OH

H2N R2

R2

O
+

+
R3

H

R4

N

C

R1

O

N
O

R3

N
H

R4

● analysis of the result (PDF-cleaving activity)○ examined by MALDI-TOF mass spectrum

J. Suh et al., J. Am. Chem. Soc. 2005, 127, 2396.
46

§3 Protein-Selective Hydrolysis of Amides

PDF-Selective Artificial Protease● property of screening
PDF (5 µM)

cat. (1.5-3 µM)○ examined by
MALDI-TOF mass spectrum

fragments

overnight
pH 7.5, 37 °C● the result of screening

H2N
O○ most active complex:

NH N
N
N
H Co OH
2 O
N OH
2
H

N
H

O
OH

J. Suh et al., J. Am. Chem. Soc. 2005, 127, 2396.
47

§3 Protein-Selective Hydrolysis of Amides

PDF-Selective Artificial Protease● cleavage site
pH 7.5, 37 ℃

○ orange: PDF / green: fragment○ C-terminal sequencing by carboxy peptidase A→ Glu152-Arg153 was the cleavage site.
● optimum pH = 7.5 / lower limit of kcat = 0.05 h-1
J. Suh et al., J. Am. Chem. Soc. 2005, 127, 2396.
48

§3 Protein-Selective Hydrolysis of Amides

PDF-Selective Artificial Protease● docking study result○ S-isomer of the catalyst with PDF
was more stable than with the
R-isomer.
○ Central acyclic chain of the catalyst
interacted with the C-terminal
α-helix, while the 3 aromatic tails
made contact with the helical andthe loop structures residing above
the active site.
J. Suh et al., J. Am. Chem. Soc. 2005, 127, 2396.
49§3 Protein-Selective Hydrolysis of Amides

PDF-Selective Artificial Protease● selectivity (control experiment)○ 15 other proteins were examined, but none ofthem were cleaved by the Co(III) complex.

H2N
O
NH N

N
H Co OH
2 O
N OH
2
H

N

N
H

O

Application to Drug is possible!

OH

list:

J. Suh et al., J. Am. Chem. Soc. 2005, 127, 2396.
50

§3 Protein-Selective Hydrolysis of Amides

AmPs-Selective Artificial Protease● AmPs: amyloidgenic peptides or proteins○ lacking active sites→ Conventional approaches can’t be applied.
○ related to diseases
Peptide-cleaving catalysts is suitable for their drug!
● target AmPs of this research○ amyloid β protein (Aβ): Alzheimer’s Disease○ human islet amyloid polypeptide (h-IAPP): type 2 diabetes
J. Suh et al., Chem. Soc. Rev. 2008, 38, 1949.
51

§3 Protein-Selective Hydrolysis of Amides

AmPs-Selective Artificial Protease● construction of a library of catalyst candidates○ synthesized according to the route shown below○ Aromatic moieties were
employed as auxiliary binding
components in the
combinatorial library.

J. Suh et al., Angew. Chem. 2007, 119, 7194.
52

§3 Protein-Selective Hydrolysis of Amides

AmPs-Selective Artificial Protease● property of screeningcat.
Aβ (4.0 µM)○ examined by
MALDI-TOF mass spectrum

fragments
pH 7.5, 37 °C
24 h● 4 complexes were selected.

Aβ40: active
Aβ42: active

Aβ40: active
Aβ42: active

Aβ40: inactive
Aβ42: active
53

Aβ40: inactive
Aβ42: active

J. Suh et al., Angew. Chem. 2007, 119, 7194.

§3 Protein-Selective Hydrolysis of Amides

AmPs-Selective Artificial Protease● property of screeningcat.
h-IAPP (4.0 µM)

fragments
pH 7.5, 37 °C
24 h

54

○ examined by
MALDI-TOF mass spectrum

J. Suh et al., J. Biol. Inorg. Chem. 2008, 13, 693.

§3 Protein-Selective Hydrolysis of Amides

AmPs-Selective Artificial Protease● 8 complexes were selected.
Aβ40: active
Aβ42: active
h-IAPP: active

Aβ40: active
Aβ42: active
h-IAPP: active

Aβ40: inactive
Aβ42: active
h-IAPP: active

Aβ40: inactive
Aβ42: inactive
h-IAPP: active

Aβ40: inactive
Aβ42: inactive
h-IAPP: active

Aβ40: inactive
Aβ42: inactive
h-IAPP: active

55

Aβ40: inactive
Aβ42: active
h-IAPP: active

Aβ40: inactive
Aβ42: inactive
h-IAPP: active

J. Suh et al., J. Biol. Inorg. Chem. 2008, 13, 693.

§3 Protein-Selective Hydrolysis of Amides

AmPs-Selective Artificial Protease● selectivity (control experiment)
horse heart myoglobin, bovine serum γ-globulin,
bovine serum albumin, human serum albumin,chicken egg white lysozyme, chicken egg ovalbumin,
bovine pancreas insulin○ Above 7 proteins were incubated with from A to H,
but protein cleavage was not observed.

Application to Drug is possible!
(Drug for all AmPs may be possible...?)
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J. Suh et al., J. Biol. Inorg. Chem. 2008, 13, 693.

§3 Protein-Selective Hydrolysis of Amides

Summary of This Section● Mb- or PDF-selective artificial protease● AmPs-selective artificial protease○ Conventional methods can’t get these fruits.

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J. Suh et al., J. Biol. Inorg. Chem. 2008, 13, 693.