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BOP Mecanismo PDF
BOP Mecanismo PDF
BOP Mecanismo PDF
Keith Proinsias
8th February 2010
Introduction
Amide bond and basic amide synthesis
Solution phase peptide synthesis
Protecting groups required for peptide synthesis
Coupling reagents used in peptide synthesis
Solid phase peptide synthesis
O
N
H
N
H
COOH
H2N
R'
-H20
H
N
R
R
O
Basic Synthesis
At the beginning acid chlorides where used.
O
R
OH
SO2Cl
+ H2N
R
Cl
R'
base
N
H
R'
+ HCl
COOH
Acid Protection
R
H2N
COOPG
R'
Coupling
PGHN
COOH
R'
R'
H
N
PGHN
COOPG
H
N
H2N
O
Dimer
COOH
R
Deprotection
R'
H
N
H2N
O
COOPG
R
R'
Coupling
PGHN
COOH
O
PGHN
R
R'
N
H
H
N
O
COOPG
R
H2N
R
R'
N
H
O
Triemer
H
N
COOH
R
Protecting Groups
Acid Protection
R
OH
H 2N
O
Protecting Groups
The most common acid protecting group used is the methyl ester.
It is stable in most coupling reaction and deprotection reaction
conditions.
Difficult to selectively remove.
H2N
Depending on what type of coupling reaction and with what amino acids
will be used other acid protecting groups can be used, such as the allyl
ester.
O
H2N
O
Protecting Groups
Amine Protection
R
OH
H 2N
O
Protecting Groups
There are two standard types of N-protecting groups used, the
Boc and Fmoc group.
O H C
3
O
CH3
N-Boc cleavage
CH3
BocHN
H 2N
COOH
COOH
Glycine
N-Fmoc cleavage
O
O
FmocHN
COOH
H2N
COOH
Glycine
Protecting Groups
Other protecting groups that can be used are Cbz and more recently
the Nosyl group.
O
O
CBZ
Leggio, A.; Gioia, M.L.D.; Perri, F.; Liguori, A. Tetrahedron, 2007, 63, 8164-8173
Protecting Groups
Protection of the R-group
R
OH
H 2N
O
Protecting Groups
Some of the different R-groups that must be protected before coupling
are hydroxyl groups (Ser), thiol groups (Cys), amines (Lys) and
carboxylic acids (Asp).
COOH
HO
COOH
HS
NH2
NH2
Serine (Ser)
H2N
Cysteine (Cys)
COOH
COOH
HOOC
NH2
Lysine (Lys)
NH2
Protecting Groups
Base Sensitive Protecting Groups
Used in N-Boc protected peptide synthesis.
Ph
COOH
Si
Ph
TBDPS
COOH
NH2
NH2
Fm
O
O
Fmoc
COOH
NH2
COOH
O
NH2
Fm
Greene, T.W.; Wuts, P.G.M. Protecting groups in organic synthesis, Fourth edition, Wiley-interscience, New York, 2006.
Protecting Groups
Acid Sensitive Protecting Groups
Used in N-Fmoc protected peptide synthesis.
Normally the t-Butyl, Boc or Trityl group is used.
Cleaved using 5-50% TFA in DCM.
Ph
COOH
COOH
Ph
O
Ph
t-Butyl
Trityl
COOH
O
t-Butyl
NH2
NH2
H
N
NH2
COOH
NH2
Boc
Greene, T.W.; Wuts, P.G.M. Protecting groups in organic synthesis, Fourth edition, Wiley-interscience, New York, 2006.
Protecting Groups
Other Protecting Groups
O
Allyl
Alloc
Cleavage: Pd(PPh3)4
O
NO2
O
S
O
Nosyl
Cleavage: Thiophenol
PMB
Greene, T.W.; Wuts, P.G.M. Protecting groups in organic synthesis, Fourth edition, Wiley-interscience, New York, 2006.
Protecting Groups
There are some exception were an unprotected amino
acid, such as serine, can be used without being
protected.
Rothman, D.M.; Vazquez, M.E.; Vogel, E.M.; Imperiali, B., Org. Lett., 2002, 4, 2865-2868
Coupling Reagents
N
N
N,N-Dicyclohexylcarbodimide (DCC)
N
N
N-(3-Dimethylaminopropyl)-N-ethylcarbonate (EDC)
DMAP
Coupling Reagents
Coupling Reagents
Racemisation of an activated amino acid
H
PG
PG
PG
N
R
O
X
O
O
O
base
X= Activator
PG
PG
PG
N
R
O
O
N
R
O
O
O
O
Coupling Reagents
N
N
N
N
N
OH
HOBt
HOAt
PF6 (BF4 )
OH
Me2N
NMe2
N
PF6
BOP
N P
PF6
Br
N P
O
N
PF6
N
PyBrOP
PyBOP
PF6
NMe2
Me2N
HATU
HBTU (TBTU)
N P
Coupling Reagents
Mechanism of HBTU Coupling
N
N
N
O
N
R
NMe2
Me2N
NMe2
NMe2
N
N
N
O
N
H
R' +
HO
N
N
R'-NH
N
N
+
Me2N
NMe2
Coupling reagents
BOP coupling reagent
Coupling reagents
PyBrOP coupling reagent
Coupling reagents
Synthesis of new coupling reagents
Coupling reagents
Synthesis of new coupling reagents
Vs
Solution Phase
Advantages
Fast production of long peptides by
increasing the amount of reactant.
Quick purification by filtration.
Automated or manual option.
Disadvantages
Expensive resin and can require
specialised equipment.
Limited scale-up.
Chan, W.; White, P. Fmoc Solid Phase Peptide Synthesis, Oxford, New York, 2000.
The resin is not completely spherical, with the reaction only occur on
the surface of the resin.
There are cavities were the coupling takes place and is the reason
why swelling is very important before coupling can occur.
Types of Resin.
You buy commercially available pre-loaded resins.
Different resins can be cleaved under basic (N-Boc) or acidic
(N-Fmoc) conditions.
Depending on the resin the final peptide can have an amide or
acid C-terminal.
Vacuum
1.
2.
3.
4.
5.
6.
7.
8.
Chan, W.; White, P. Fmoc Solid Phase Peptide Synthesis, Oxford, New York, 2000.