semisynthetic enzymes for organic synthesis 赵峰 2014.3.22 1
TRANSCRIPT
Semisynthetic Enzymes for Organic Synthesis
赵峰
2014.3.22
1
Introduction
Homogeneous Enzymatic
Enantiomers easy access challenging
Solvent tolerance mostly organic mostly aqueous
Substrate scope large narrow
Optimization chemical genetic
Turnover number limited large
Metals involved any metal limited (biorelevant)
Features of Homogeneous and Enzymatic Catalysis
Ward, T. R. Acc. Chem. Res. 2011, 44, 47-57 2
Combination——Semisynthetic Enzymes
Mcat
M=metal M=molecue
Mcat
For Selectivity Mcat For Catalytic Activity
3
Content
• Artificial Metalloenzymes– Covalent Strategies and applications
– Supramolecular Strategies and applications
• Nonmetal-Containing Systems– Covalent Strategies and applications
• Conclusion
4
Content
• Artificial Metalloenzymes– Covalent Strategies and applications
– Supramolecular Strategies and applications
• Nonmetal-Containing Systems– Covalent Strategies and applications
• Conclusion
5
Mcat
First coordination sphere:defines Mcat reactivity
Scend coordination sphere:substrate binding, steric, electronic, hydrophobic,H-bonding, and othereffects conveyed to Mcat
Vacant space forsubstrates andcatalyst (Mcat)
M=metal
XY
Mcat Mcat
Colvalent anchoring Supramolecular anchoring
protect Mcat,scaffold binding, etc.
General Structure of Artificial Metalloenzymes
6
Covalent Anchoring Strategies
R SH
cysteine
X Y X Y
R' Br
R'X
O
R'S
SOO
NO
O
R'
N
N
R'
O
N
O
O
NHNH2
O
then
R' CHO
R S
R S
R S
R S
R S
R S
R'
R'
O
SR'
NO
O
R'
R'
O
N
O
O
NHN
O
R'2
2
H2NHC C OH
CH2
SH
O
Lewis, J. C. ACS Catal. 2013, 3, 2954-2975
XY
McatX
YMcat
Conditions
7
Covalent Anchoring Strategies
R NH2
lysine
X Y X Y
R OH
RN3
serine
p-azidophenylalanine
S NH2then R'
X
OR
HN
S
NH2
R'
O
3
R'P pNP
O
OAk/pNP
R OP
O
R'OAk/pNP
R' R'N
NN
R
H2NOH
O
NH2
HO OH
O
NH2
Ph OH
O
NH2
Lewis, J. C. ACS Catal. 2013, 3, 2954-29758
Representative Cofactors
N N
HNI
O
N
O
O
O
O
2
N N
t-Bu
t-Bu
O OMn
Cl
N
N
NM
OO
N N
O OMn
Cl
SX
O
XS
X=SO2Me
N N
Ru
NH
Br
MesMes
i-PrO
ClCl
O
Ph2PN
P
O O
pNPpNP32
P
O
EtOPNp
L
L
ML MNMe2 PtClSPh PdBrSMe PdBr
N
N
NM
O O
OH
HO
O
O
OO
OO
Rh Rh
O
O
OO
9
Reactions by Covalent Anchoring AME
10
N
N
NH
OI
ALBP-SH
N
N
NH
OSALBP
85%-95%
ALBP-Phen
H2O
PIPES buffer, PH 6.1, 25C
ALBP-Phen-Cu(II)
ROR'
O
NH2
ROH
O
NH2
+ R' OH
Distefano,M. D. J. Am. Chem. Soc. 1997 , 119, 11643-11652
N
NCuH2N
OR
OR'HO-
N
NCuH2N
OR
O+ R'OH
Reactions by Covalent Anchoring AME
11Hilvert, D. Chem. Commun., 2011, 47, 12068–12070
N
Ts catalyst
45C, 12hN Ts
12
Reactions by Covalent Anchoring AME
Roelfes, G. Angew. Chem. Int. Ed. 2012, 51, 7472 –7475
N
O
R
LmrR-1-Cu (3mol%)
NO
R
R LmrR variant conv(%) ee(%) endo/exoPh M89C, V15A 89 97 96:4m-MeOPh M89C 56 93 96:4Me M89C 97 <5
13
Reactions by Covalent Anchoring AME
N
O
R
LmrR-1-Cu (3mol%)
N
O
R LmrR variant conv(%) ee(%) i-Pr M89C 67 77 t-Bu M89C 80 84 n-pent M89C 57 67i-Pr M89C,D100A 28 <5
R
OH
Roelfes, G . Chem. Sci., 2013, 4, 3578–3582
Content
• Metal-Containing Systems– Covalent Strategies and applications
– Supramolecular Strategies and applications
• Nonmetal-Containing Systems– Covalent Strategies and applications
• Conclusion
14
Supramolecular Anchoring Strategies
15
Mcat
Mcat
binding to substituent
OH
NHO
H
HN
H S
O
Bitotin
Take advantage of the remarkable affinity of biotin for either avidin or streptavidin！
Represetative Biotinylated Cofactors
NH
O
HHNH
SR
O
N PPh2
RhL2
Ph2P
Ph2P
PPh2
Rh(COD)+
NH O
NH
PPh2
PPh2
Rh(COD)+
N PPh2
Rh(COD)+
Ph2P
O
NH
NH
SN M
NH
OO
Cl
n-CnRn n-CnRn
Ir 5-C5R5
Rh 5-C5R5
Ru 6-C6R6
Ru 6-p-cymene
N
N
Ru
Mes
MesOi-Pr
ClCl
NH
L=NBD, COD
16
COOH
NHCOCH3
1.5 atm H2, Avidin, Rh-complex
CH3CHCOOH
NHCOCH3
44%ee(S)
0.1 M Na2PO4 buffer, pH=70C, 48h
Whitesides, G. M. J. Am. Chem. Soc. 1978, 100, 306–307 17
NH
O
HHNH
SCN
OPh2P
PPh2
Rh
Tf
Rh-complex
Protein TONee%
Polarimetric NMR
None 475 <2 <2
Lysozyme 450 <1 <2
CA 50 <10
BSA 150 <5
Avidin(1equiv)
>500 41 44
Avidin.biotin 200 <4
Reactions by AME Bsed on Biotin-Avidin Technology
18
Reactions by AME Bsed on Biotin-Avidin Technology
COOH
NHCOCH3
1.5 atm H2, Rh-complex,CH3CH
COOH
NHCOCH3
(Strept)avidin 92%ee(R)(Strept)avidin S112G 96%ee(R)
0.1 M Na2PO4 buffer, pH=70C, 48h
Ward, T. R. J. Am. Chem. Soc. 2003, 125, 9030-9031
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Reactions by AME Bsed on Biotin-Avidin Technology
NH
O
HHNH
S
HN
O
S
HN
OO
H2N
n-(CnRn)MCl]
NEt3, i-PrOH, refluxNH
O
HHNH
S
HN
O
SN
OO
NH2MCl
n-(CnRn)
Biot-q-LH n-(CnRn)M(Biot-q-L)Cl
n-(CnRn)
Ward, T. R. J. Am. Chem. Soc. 2006, 128, 8320-8328
20
Reactions by AME Bsed on Biotin-Avidin Technology
Ward, T. R. Angew. Chem. Int. Ed. 2011, 50, 3026 –3029
21
Reactions by AME Bsed on Biotin-Avidin Technology
Ward, T. R. Angew. Chem., Int. Ed. 2008, 47, 701–705
22
Reactions by AME Bsed on Biotin-Avidin Technology
Ward, T. R. Chem. Commun. 2011, 47, 12065-12067
Content
• Artificial Metalloenzymes– Covalent Strategies and applications
– Supramolecular Strategies and applications
• Nonmetal-Containing Systems– Covalent Strategies and applications
• Conclusion
23
Scend coordination sphere:substrate binding, steric, electronic, hydrophobic,H-bonding, and othereffects conveyed to Mcat
Vacant space forsubstrates andcatalyst (Mcat)
XY
Mcat Mcat
Colvalent anchoring Supramolecular anchoring(few reports)
protect Mcat,scaffold binding, etc.
M=molecue
Mcat
General Structure of Nonmetal-Containing Systems
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Colvalent Anchoring Strategy
NH3+ SO3
-
NMe3+
ONH3
+
NH3+
NH3+
NH3+
COOH
HOOC COOH
COOH
COOHHOOC OHHO
H
H
OH
COOH
COOH
Subtilisin SH SCH3SO2SSR
Subtilisin S RpH 9.5
R=
Jones, J. B. J. Am. Chem. Soc. 1997, 119, 5265-5266
Jones, J. B. Bioorg. med. Chem.. 1999, 7, 1381 25
Colvalent Anchoring Strategy
N
SS
N
NH2
OH
ALBP SH +
NH
S
S
N
NH2
OH
ALBP S
+
Distefano, M. D. J. Am. Chem. Soc., 1996, 118, 10702-10706
N
N
NH
N
H3C
Br
O
O
R
Papain SH +
N
N
NH
N
H3C
O
O
R
Papain S
R=CH2(CHOAc)3CH2OAc, CH3
Kaiser, E. T. Biochem. Biophys. Res. Commun. 1977, 76, 64-70
Subtilisin OHPMSF
Subtilisin OSO2CH2PhNaSeH
Subtilisin SeH
Hilvert, D. J. Am. Chem. Soc. 1989, 111, 4513-4514
Papain SH + Papain S
R=C6H5CH2, CH3
N
S
BrR
N
S
R
Suckling, C. J. Bioorg. Med. Chem. 1993, 3, 531-534
26
N
N
NH
N
H3C
Br
O
O
R
Papain SH +
N
N
NH
N
H3C
O
O
R
Papain S
R=CH2(CHOAc)3CH2OAc, CH3
N
N
NH
N
O
O
CH3
N
N
NH
N
O
O
CH3
N
N
NH
N
O
O
CH3
Br
O
Br
O BrO
Kaiser, E. T. Biochem. Biophys. Res. Commun. 1977, 76, 64-7027
Reactions by Nonmetal Artificial Enzymes
N
R
HH
NH2
OFlavopapains
O2 N
R
NH2
OH
Reactions by Nonmetal Artificial Enzymes
Papain SH + Papain S
R=C6H5CH2, CH3
N
S
BrR
N
S
R
O O O+
OO
O OH
Artificial enzyme
28%yield
60%yield15%yield no enzyme
Suckling, C. J. Bioorg. Med. Chem. 1993, 3, 531-534
28
Reactions by Nonmetal Artificial Enzymes
Subtilisin OHPMSF
Subtilisin OSO2CH2PhNaSeH
Subtilisin SeH
Hilvert, D. J. Am. Chem. Soc. 1989, 111, 4513-451429
N
O
+ Subtilisin-SeH
Se-Subtilisin
O
H2Obutylamine
NH
O
C4H9
OH
O
Reactions by Nonmetal Artificial Enzymes
30
N
SS
N
NH2
OH
ALBP SH +
NH
S
S
N
NH2
OH
ALBP S
+
N
R'NH2
OH+
R
O
O
OH
N
R'O
OH+
R
NH2
O
OH
R'=Protein
0~94%ee
Distefano, M. D. J. Am. Chem. Soc., 1996, 118, 10702-10706
Content
• Artificial Metalloenzymes– Covalent Strategies and applications
– Supramolecular Strategies and applications
• Nonmetal-Containing Systems– Covalent Strategies and applications
• Conclusion
31
Conclusion• A lot of AMEs have been synthesized, and some of them have
been successfully applied to organic synthesis.
• Construction of AMEs can be complicated to get host Protein and guest motif ready for conjunction at the same time
• The reaction scope is still narrow, and selectivity is not very good
• Further research work is desired to enlarge the number of AME family and find new applications of them
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