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1. Occurrence and Functions of Peptides in Nature and Every Day Life …hormones, neurotransmitters, therapeutics, artificial sweetener, … 2. Peptide Synthesis a) Aspartam: Properties of amino acids; nomenclature; solution phase synthesis b) Glucagon: Solid phase peptide synthesis c) Fuzeon, an anti HIV drug: Solution and solid phase peptide synthesis 3. Peptide Structures – Nanostructured Materials – Self-assembly a) Fuzeon: α-helices and coiled-coil structures b) Amyloids: β-sheets c) Collagen: PPII helices 4. Applications of Peptides in Chemistry, Biology and Material Sciences a) Peptide-based materials b) Therapeutically active peptides b) Cyclic peptides and cancer imaging c) Asymmetric Catalysis with Peptides – Combinatorial chemistry
Syllabus
13 Prof. H. Wennemers
Solid Phase Peptide Synthesis (SPPS) – Fmoc/tBu Strategie in the example: Wang Linker
O
ClR1
CO2FmocHN Cs LinkerOFmocHNO
R1
LinkerOH2NO
R1
LinkerONH O
R1OFmocHN
R2
DMF
Piperidine:DMF 1:4 (1x 3 min, 1x 7 min)
R2
CO2HFmocHN3 equiv.
3 eq. PyBOP, 3 eq. iPr2NEt
repeat Fmoc deprotection/AA couplings to desired peptide length
LinkerONH O
R1OHN
R2OH2N
RnOHN
H O
R1´OHN
R2´OH3N
Rn´95% TFA
(scavengers)
side-chain deprotected peptide
Wang Linker
14 Prof. H. Wennemers
Kupplungsreagenzien
N C NR
R
Carbodiimide R = R´ = cyclohexyl: N,N´-Di-Cyclohexyl Carbodiimid (DCC) R = R´ = iso-propyl: N,N´-Di-Isopropyl Carbodiimid (DIC) R = Et, R´= CH2CH2CH2NMe2 : N-Ethyl-N´-(3-Dimethylaminopropyl) Carbodiimid (EDC)
Phosphonium Reagenzien Guanidinium/Uronium Reagenzien
NN
NO P
N
NN
PF6
NN
NO P
NMe2
NMe2NMe2
PF6
NN
N
OO
P OEt
O
OEt
NN
N PF6
NN
ON N
NN PF6
NN
O
NN
N BF4
NN
O
HBTU HATU
TBTU
BOP (Castro`s Reagent)
PyBOP
DEPBT
15 Prof. H. Wennemers
Commonly used acid sensitive protecting groups for the functional groups of the amino acid side chains
Fmoc: α-NH group Boc: Lysine, Tryptophan, Histidine Pbf: Arginine Trt: Cysteine, Aspartic acid, Glutamic acid Acm: Cysteine tBu: Serine, Threonine, Hydroxyproline, Tyrosine
Prof. H. Wennemers 16
Linker für die Festphasensynthese
O
OR
O
O
OR
O OMe
O
Ph
R
O
Cl
O
HN
R
OMeO
MeO
NH
OOR
O
NO2
O
O
R
O
HN
O
S NH
O
NH
R
O OO
Wang SASRIN® Chlorotrityl
Rink Amide Hycram
o-Nitrobenzyl Kenner Safety Catch
17 Prof. H. Wennemers
Peptidsynthese an fester Phase (Nobelpreis für Chemie 1984 für Bruce Merrifield)
Peptidsynthesizer 1966 heute
18 Prof. H. Wennemers
BACHEM Holding AG │ Company Presentation Seite 19 │ September 2012
Insulin (Insulin Superfamily) inac*ve hexameric form – bound to Zn2+:
Insulin and Glucagon
Glucagon (Secre3n Family)
19
BACHEM Holding AG │ Company Presentation Seite 20 │ September 2012
Comparison: Solution vs. Solid Phase
Calcitonin Glucagon - Produced since 1986 1996 - Synthetic steps 104 56 - Purification CCD
two-dim. HPLC three-dim. HPLC - Derivatives needed 158 kg 29 kg
for the production of 1kg - Time for completion 2.5 years 0.6 years
Human Parathyroid Hormone (1-84)
Parathyroid Hormone (PTH)
T. Michels, R. Dölling, U. Haberkorn, W. Mier, Org. Le(., 2012 , 14, 5218-‐5221. R. Balling, R. G. Erben, Nature Medicine, 2000, 6, 860-‐861.
PTH fragment (46-‐84): AGSQRPRKKEDNVLVESHEKSLGEADKADVNVLTKAKSQ-‐NH2
addition of 5% formic acid to the piperidine
21
Aspartimid Formation
D. Housset, C. Habersetzer-‐Rochat, J.-‐P. As3er, J. C. Fontecilla-‐Camps, J. Mol. Biol., 1994, 238, 88–103.M. Mergler, F. Dick, B. Sax, P. Weiler, T. Vorherr, J. Pep>de Sci. 2003, 9, 36–46. T. Michels, R. Dölling, U. Haberkorn, W. Mier, Org. Le(., 2012 , 14, 5218-‐5221.
R NH
HN
NH
O
O
O
O
R1
R2
OtBu
O
R NH
NNH
O
O
OO
R1
R2
O
R NH
HN
NH
O
O
O
O
R1
R2
OH
O
R NH
HN
NH
O
O
O
O
R1
R2
NO
HN
NHO
O
O
R1
R2
HN
O
R
OHO
HN
NHO
O
O
R1
R2
HN
O
R
NO
Acid or Base(epimerisation)
Hydrolysis(epimerisation)
20% Piperidine/DMF(epimerisation)
α-PeptidePiperidide of α-Peptide
22
Side-reactions of Met
Y. S. Kim, J. A. Moss, K. D. Janda, J. Org. Chem. 2004, 69, 7776-7778.D. Picone et al, ChemBioChem 2006, 7, 257 – 267.T. Lührs, C. Ritter, M. Adrian, D. Riek-Loher, B. Bohrmann, H. Döbeli, D. Schubert, R. Riek, PNAS 2005, 102, 48, 17342–17347.
Aqueous Solution Structure of the Alzheimer's Disease Aβ Peptide 3D Structure of Alzheimer's Aβ(1-42) fibrils
23
