1. TWEEZER PEPTIDE MIMICS
OF THE ESTROGEN
RECEPTOR FOR
DETERMINATION OF
ENDOCRINE DISRUPTOR
CHEMICALS
Rosa Romeralo-Tapia
Supervisor: Prof. Dr. Johan Van der Eycken
Laboratory for Organic and Bioorganic SynthesisThesis submitted to obtain the degree of
Doctor in Sciences: Chemistry
13 October 2011
4. • Challenges in endocrine disrupting chemicals
research: EST-SENDICHEM project
• Design artificial receptor
• Synthesis type A, B and C tweezer peptide
mimics
4/75
Overview
5. • Challenges in endocrine disrupting chemicals
research: EST-SENDICHEM project
• Design artificial receptor
• Synthesis type A, B and C tweezer peptide
mimics
• Structure analysis by NMR spectroscopy
5/75
Overview
6. • Challenges in endocrine disrupting chemicals
research: EST-SENDICHEM project
• Design artificial receptor
• Synthesis type A, B and C tweezer peptide
mimics
• Structure analysis by NMR spectroscopy
• Screening tweezer peptide mimics by ACE6/75
Overview
8. “Endocrine disruptors are defined as exogenous substances or
mixtures that alter the function(s) of the endocrine system and
consequently cause adverse health effects in intact organisms, or their
progeny or (sub) population”*
Definition of EDCs
NATURAL and SYNTHETIC
HORMONES
COOLANTS, PESTICIDES PLASTICISERS
HO
OH
OH
Cl
Cl
Cl
Cl
Cl
O
O
O
O
ClCl
HO
OH
Cl
Cl Cl
Estradiol (E2) Polychlorinated bisphenols
(PCBs)
Bisphenol-A (BPA)
Ethinylestradiol (EE) 1,1,1-Trichloro-2,2-bis-
(4'-chlorophenyl)ethane (DDT)
Di-n-butylphthalate
(DBP)
HO
*Definition given by International Programme on Chemical Safety
8/75
10. Challenges in EDC Research
• Establish cause-effect
relationships
• Obtain reliable exposure data
• Identify which chemicals cause
endocrine disruption
10/75
11. Challenges in EDC Research
• Establish cause-effect
relationships
• Obtain reliable exposure data
• Identify which chemicals cause
endocrine disruption
11/75
12. NEW SORBENT TO
SELECTIVELY
RETAiN EDCs FROM WATER
EDC-CONTAMINATED
WATER
Solid phase affinity extraction
Prof. dr. Annemieke Madder
(Coordinator)
Organic and Biomimetic Chemistry
Prof. dr. P. Sandra
Separation Sciences
Prof. dr. F. Du Prez
Polymer Chemistry
Prof. dr. J. Van der Eycken
Organic and Bioorganic Synthesis
Prof. dr. J. Martins
NMR & Structure Analysis
12/75
13. Mode of action of
EDCs
Hormone-related
mechanisms
mone synthesis (1)
mone transport (2, 3, 11)
mone metabolism (4)
www.medscape.com Source: Expert Rev Of Obstet Gynacol 2008
13/75
14. Mode of action of
EDCs
B) Receptor-mediated
mechanisms
Hormone-related
mechanisms
mone synthesis (1)
mone transport (2, 3, 11)
mone metabolism (4)
www.medscape.com Source: Expert Rev Of Obstet Gynacol 2008
14/75
15. ://whqlibdoc.who.int
B) Receptor mediated EDCs GENISTEIN (natural hormone)
ETHINYLESTRADIOL, DIETHYLSTILBESTROL (synthetic
hormones)
DDT (pesticide), BISPHENOL-A (plasticiser)
• Agonist: mimicking the natural hormone ER
recognizes
• Antagonist: blocking the hormone receptor positions EDCs as
ligands
Mode of action of
EDCs
Natural
hormone
EDC
ER
ER Estrogen
Receptor
15/75
16. NEW SORBENT TO
SELECTIVELY
RETAiN EDCs FROM WATER
(based on receptor-mediated
mechanism)
EDC-CONTAMINATED
WATER
Solid phase affinity extraction
Van der Plas SE et al. Eur. J. Org. Chem. 2009, 11 (Sp. Iss. SI), 1796-1805
Figaroli S, Madder A. Tetrahedron, 2010, 66, 6912-6918
Solid support (Polymer)
Molecule with affinity to
EDCs:
MIMIC OF
ESTROGEN
RECEPTOR
16/75
20. Histidine 524 (H11), Arginine 394 (H5), Glutamic acid 353 (H3)
hERα-E2
Hydrophilic interactions
Design HBD-hERα mimic
Amino acids
OH
HO
Estradiol (E2)
Tanenbaum D.M et al. Proc. Natl. Acad. Sci. USA 1998, 95, 5998-6003
20/75
21. A-B rings: Ala 350 (H3), Leu 387 (H5), Leu 391 (H5), Phe 404
(βturn), Met 388 (H5) D ring: Leu 525 (H11), Ile 424 (H7), Gly 521
(H11)
hERα-E2
Hydrophobic residues
Design HBD-hERα mimic
Amino acids
Tanenbaum D.M et al. Proc. Natl. Acad. Sci. USA 1998, 95, 5998-6003
21/75
22. DES
Hydrophilic
contacts
Glu 353
Arg 394
His 524
Hydrophobi
c
contacts
Ala 350
Leu 386
Leu 387
Raloxifene
Hydrophilic
contacts
Glu 353
Arg 394
His 524
Hydrophobi
c
contacts
Ala 350
Leu 354
Leu 387
E2
Hydrophilic
contacts
Glu 353
Arg 394
His 524
Hydrophobi
c
contacts
Ala 350
Leu 387
Met 388
Leu 391
Design HBD-hERα mimic
Amino acids
Glu
Arg
His
Ala
Leu
Met
Phe
Data obtained from Protein Data Bank http:// www.pbd.org
DES: diethylstilbestrol E2: 17Beta-estradiol
22/75
23. .H1H2..MGLLTNLADREL..H4..LEILMIGLVWR.. βturn
..H6H7H8H9H10..GMEHL..
Helix 3 Helix 5
Helix 11
H12..C terminal
His 524
(H11)
Arg 394 (H5)
Glu 353 (H3)
Ala 350 (H3)
Leu 387 (H5)
Leu 391 (H5)
Phe 404 (βturn)
Met 388 (H5)
Leu 525 (H11)
Ile 424 (H7)
Gly 521 (H11)
N
C
H11
H3
H5
H7
Design HBD-hERα mimic
Tertiary structure
N C
C
23/75
34. O
O
NO2
NHFmoc
O
O
NO2
HN
O
NHFmoc
Wang
resin
l = 1.10 mmol/g
(i) 20% piperidine/DMF
(ii) Fmoc-Gly-OH, DIC, DMAP,
CH2Cl2 (x2)
Preactivation
reagents
(1st coupling)
Preactivation
reagents
(2nd
coupling)
Loading
(mmol/g)
Yield
(%)
Fmoc-Gly-OH,
DIC
Fmoc-Gly-OH,
DIC, HOBt
0.58 57%
Fmoc-Gly-OH,
DIC, HOBt
Fmoc-Gly-OH,
DIC, DMAP
0.90 89%
Fmoc-Gly-OH,
DIC, DMAP
Fmoc-Gly-OH,
DIC, DMAP
1.00 Quantitative
Type A model: Linear
SPPS
34/75
35. O
O
NO2
HN
O
N
H
O
H
N
N
H
O
O
H
N O
O
O
NO2
HN
O
NHFmoc
1. 20% piperidine/DMF
2. Fmoc-Ala-OH, HBTU, DIPEA, DMF
3. 20% piperidine/DMF
4. Fmoc-Leu-OH, HBTU, DIPEA, DMF
5. 20% piperidine/DMF
6. Fmoc-Phe-OH, HBTU, DIPEA, DMF
7. 20% piperidine/DMF
8. DIPEA/Ac2O/CH2Cl2 (1/1/3)
Type A model: Linear
SPPS
35/75
36. O
O
NH2
HN
O
N
H
O
H
N
N
H
O
O
H
N O
O
O
NO2
HN
O
N
H
O
H
N
N
H
O
O
H
N O
V.2
V.3
SnCl2. 2H2O
DMF
Crude cleaved product
Crude cleaved product
min0 5 10 15 20
mAU
0
50
100
150
200
250
300
350
11.259 min
min0 5 10 15 20
mAU
0
200
400
600
800
1000
1200
DAD1 B, Sig=254,20 Ref=off (F:07-11-06056-1201.D)
10.449 min
min0 5 10 15 20
mAU
0
200
400
600
800
1000
1200
)
10.449 min
Type A model: Linear
SPPS
36/75
Solvent A: 5mM NH4OAc/H2O Solvent B: CH3CN 0-100% B in 15 minutes (λ = 214 nm )
37. O
O
O
NH2
HN
O
N
H
O
H
N
N
H
O
O
H
N
1. Fmoc-Gly-OH, DIC, DMAP, CH2Cl2 (x 2)
2. 20% piperidine/DMF
3. Fmoc-Glu(tBu)-OH, HBTU, DIPEA, DMF (x 2)
4. 20% piperidine/DMF
Repeat 3,4
5. Fmoc-Ala-OH, HBTU, DIPEA, NMP
Repeat 4
6. DIPEA/Ac2O/CH2Cl2 (1/1/3) (x 2)
7. 95%TFA/H20
HO
O
NH
O
H
N
NH
N
H
H
N
N
H
N
H
H
N
O
O
O
O
O
H
N
N
H
O
O
OHO
O
O
Type A model: Linear
SPPS
37/75
38. Type B and C
Model Peptide Mimics
based on Convergent
Cu-Catalyzed 1,3 Dipolar
Cycloaddition Reactions
38/75
40. Franke R et al. Tetrahedron Lett. 2005, 46, 4479-4482
CuAAC
Cu-alkyne azide 1,3-cycloaddition
H2N
R1
N
N
N
CO2H
R2
H2N
N
O
R1
CO2H
R2
1
23
4
H
H acceptor H acceptor
H
H donor H donor
AMIDE
BOND
1,4-SUBSTITUTED
TRIAZOLE
RING
i) N3CH2CO-TSKYREG-OH,
CuI
ii) Cleavage
1
4
40/75
45. Synthesis
Azidopeptides
O
O
N
H
O
NH2 O
O
N
H
O H
N
N3
O
R1
R2HBTU, DIPEA,
DMF (x 2)
2-ClTrt.
resin
R1
R2
HBTU-mediated
Fmoc SPPS
AcOH/ TFE/ CH2Cl2
O
N3
HO
23-70% crude purity
HO
O
N
H
O H
N
N3
O
R1
R2
Azido acetic acid
coupling to free amine
min0 5 10 15 20
mAU
0
200
400
600
800
08-12-11 -
10.502
10.941
11.524
min0 5 10 15 20
mAU
0
200
400
600
800
- -
10.502
10.941
N3CH2CO-Leu-Phe
Leu-Phe
HOBt
Synthesis N CH CO H:Franke R et al. Tetrahedron Lett. 2005, 46, 4479-4482
45/75
46. Imidazole-1-sulfonyl azide hydrochloride: Goddard-Borger ED et al. Org. Lett. 2007, 9, 3793-3800
Devulder V, Backaert F, Van der Eycken J. Ghent University. 2010
O
O
N
H
O H
N O
O
N
H
O H
N
N3
O
R1
R22-Cl Trt.
resin
R1
R2
HBTU-mediated
Fmoc SPPS
AcOH/ TFE/ CH2Cl2
CuSO4. 5H2O
THF/H2O
R3
O
NH2
R3
N
N
S
O
N3
O
HCl
OH
O
N
H
O
H
N
N3
O
R1
R2
R3
58-76% crude purity
Synthesis
AzidopeptidesDiazo transfer reaction
min0 5 10 15 20
mAU
0
200
400
600
800
1000
N3Leu-Leu-Leu-Gly-Leu-Phe-OH
N3Leu-Leu-Gly-Leu-Phe-OH
Leu-Leu-Leu-Gly-Leu-Phe-OH
46/75
48. O
NHFmoc
O H
N
O
N
N N
O
H
N
O
N
H
O
OH
i) 20%piperidine/DMF
ii) 4-pentynoic acid,
HBTU, DIPEA, DMF
O
NHFmoc
O H
N
O
HO
O
N
H
O H
N
N3
O
CuI, Ascorbic acid, DIPEA,
2,6-lutidine/DMF
NH
NH
HO
O
O N
N
H
N
H
N
O
O
H
N
O
OH
ON N
O
N
H
O
OH
ONN
HO
O
N
H
O H
N
N3
O
CuI, Ascorbic acid, DIPEA,
2,6-lutidine/DMF
i)
ii) TFA,H2O (95%)
NH
NH
O
O
O N
N
H
O
O
H
N
O
OH
ON N
Type B model: Sequential Click
48/75
New scaffold
49. O
NHFmoc
O H
N
O
N
N N
O
H
N
O
N
H
O
OH
i) 20%piperidine/DMF
ii) 4-pentynoic acid,
HBTU, DIPEA, DMF
O
NHFmoc
O H
N
O
HO
O
N
H
O H
N
N3
O
CuI, Ascorbic acid, DIPEA,
2,6-lutidine/DMF
NH
NH
HO
O
O N
N
H
N
H
N
O
O
H
N
O
OH
ON N
O
N
H
O
OH
ONN
HO
O
N
H
O H
N
N3
O
CuI, Ascorbic acid, DIPEA,
2,6-lutidine/DMF
i)
ii) TFA,H2O (95%)
NH
NH
O
O
O N
N
H
O
O
H
N
O
OH
ON N
Type B model: Sequential Click
49/75
50. O
NHFmoc
O H
N
O
N
N N
O
H
N
O
N
H
O
OH
i) 20%piperidine/DMF
ii) 4-pentynoic acid,
HBTU, DIPEA, DMF
O
NHFmoc
O H
N
O
HO
O
N
H
O H
N
N3
O
CuI, Ascorbic acid, DIPEA,
2,6-lutidine/DMF
NH
NH
HO
O
O N
N
H
N
H
N
O
O
H
N
O
OH
ON N
O
N
H
O
OH
ONN
HO
O
N
H
O H
N
N3
O
CuI, Ascorbic acid, DIPEA,
2,6-lutidine/DMF
i)
ii) TFA,H2O (95%)
NH
NH
O
O
O N
N
H
O
O
H
N
O
OH
ON N
Type B model: Sequential Click
50/75
51. O
NHFmoc
O H
N
O
N
N N
O
H
N
O
N
H
O
OH
i) 20%piperidine/DMF
ii) 4-pentynoic acid,
HBTU, DIPEA, DMF
O
NHFmoc
O H
N
O
HO
O
N
H
O H
N
N3
O
CuI, Ascorbic acid, DIPEA,
2,6-lutidine/DMF
NH
NH
HO
O
O N
N
H
N
H
N
O
O
H
N
O
OH
ON N
O
N
H
O
OH
ONN
HO
O
N
H
O H
N
N3
O
CuI, Ascorbic acid, DIPEA,
2,6-lutidine/DMF
i)
ii) TFA/ H2O (95%)
NH
NH
O
O
O N
N
H
O
O
H
N
O
OH
ON N
Type B model: Sequential Click
51/75
52. NH
NH
HO
O
O N
N
H
N
H
N
O
O
H
N
O
OH
ON N
O
N
H
O
OH
ONN
Crude product ( 7 steps)
min0 5 10 15 20
mAU
0
200
400
600
800
1000
DAD1 A, Sig=214,20 Ref=off (F:09-02-09080-1201.D)
9.225
9.686
10.149
13.340
min0 5 10 15 20
mAU
-100
-50
0
50
100
150
200
250
300
DAD1 A, Sig=214,20 Ref=off (F:09-07-02093-0501.D)
10.285
After RP-HPLC
Purity 82%
Type B model: Sequential Click
52/75
Solvent A: 5mM NH4OAc/H2O Solvent B: CH3CN 0-100% B in 15 minutes (λ = 214 nm )
54. O
NHFmoc
O H
N
O
Wang
resin
O
O
HN
O
H
N
O
N
H
O
H
N
N
H
O
O
NHFmoc
O OtBu
i) DBU/4-methylpiperidine/DMF (3/17/80)
ii)Fmoc-AA-OH, HATU, DIPEA, DMF
x 4 AA
i)
CuI, ascorbic acid, DIPEA,
2.6-lutidine/DMF, 80ºC, MW
ii) DBU/4-methylpiperidine/DMF (3/17/80)
iii) TIS/TFA/H2O
NH
NH
HO
O
O N
N
H
H
N
N
H
H
N
O
O
H
N
O
OH
ON N
NH2
O
HO O
O
O
HO
O
N
H
O H
N
N3
O
Type C model: SPPS-Click
54/75
55. O
NHFmoc
O H
N
O
Wang
resin
O
O
HN
O
H
N
O
N
H
O
H
N
N
H
O
O
NHFmoc
O OtBu
i) DBU/4-methylpiperidine/DMF (3/17/80)
ii) Fmoc-AA-OH, HATU, DIPEA, DMF
x 4 AA
i)
CuI, ascorbic acid, DIPEA,
2.6-lutidine/DMF, 80ºC, MW
ii) DBU/4-methylpiperidine/DMF (3/17/80)
iii) TIS/TFA/H2O
NH
NH
HO
O
O N
N
H
H
N
N
H
H
N
O
O
H
N
O
OH
ON N
NH2
O
HO O
O
O
HO
O
N
H
O H
N
N3
O
Type C model: SPPS-Click
55/75
56. O
NHFmoc
O H
N
O
Wang
resin
O
O
HN
O
H
N
O
N
H
O
H
N
N
H
O
O
NHFmoc
O OtBu
i) DBU/4-methylpiperidine/DMF (3/17/80)
ii)Fmoc-AA-OH, HATU, DIPEA, DMF
x 4 AA
i)
CuI, ascorbic acid, DIPEA,
2.6-lutidine/DMF, 80ºC, MW (80W)
ii) DBU/4-methylpiperidine/DMF (3/17/80)
iii) TIS/TFA/H2O
NH
NH
HO
O
O N
N
H
H
N
N
H
H
N
O
O
H
N
O
OH
ON N
NH2
O
HO O
O
O
HO
O
N
H
O H
N
N3
O
Type C model: SPPS-Click
56/75
57. O
NHFmoc
O H
N
O
Wang
resin
O
O
HN
O
H
N
O
N
H
O
H
N
N
H
O
O
NHFmoc
O OtBu
i) DBU/4-methylpiperidine/DMF (3/17/80)
ii)Fmoc-AA-OH, HATU, DIPEA, DMF
x 4 AA
i)
CuI, ascorbic acid, DIPEA,
2.6-lutidine/DMF, 80ºC, MW
ii) DBU/4-methylpiperidine/DMF (3/17/80)
iii) TIS/TFA/H2O
NH
NH
HO
O
O N
N
H
H
N
N
H
H
N
O
O
H
N
O
OH
ON N
NH2
O
HO O
O
O
HO
O
N
H
O H
N
N3
O
Purity 80%
min0 5 10 15 20
mAU
-100
-50
0
50
100
150
200
250
300
DAD1 A, Sig=214,20 Ref=off (10-01-25074-3301.D)
7.999
8.869
Type C model: SPPS-Click
57/75
Solvent A: 5mM NH4OAc/H2O Solvent B: CH3CN 0-100% B in 15 minutes (λ = 214 nm )
58. STRUCTURE ANALYSIS
BY
NMR
SPECTROSCOPY
“The biological function of peptides and
proteins is defined by their ability to adopt
well-defined conformations that complement
those of their binding partner […]”
Pedersen DS, Andrew Abell Eur. J. Org. Chem. 2011, 2399-2411
60. NH
NH
HO
O
O N
N
H
N
H
NO
O
H
N
O
OH
ON N
O
N
H O
OH
ONN
Hα
Hβ
Hβ
Hβ
γH
δH
δH
Hα
Type B peptide mimic : H-H TOCSY
700 MHz, 25°C,
CD3CN/H2O
ppm
7.27.47.67.88.08.2 ppm
9
8
7
6
5
4
3
2
1
Leu NH
Leu Hα
Leu Hβ
Leu Hγ
Leu Hδ
Leu Hδ
Phe NH
Phe Hα
Phe Hβ
Phe Hβ
In collaboration with K Gheysen, Prof dr J Martins
60/75
61. NH
NH
HO
O
O N
N
H
H
N
N
H
H
N
O
O
H
N
O
OH
ON N
NH2
O
HO O
O
CH3 O
Hα Hα
Hα
Hα
Hα
Hα
Hβ
Hβ
Gly1
Gly2
β
700 MHz, 25°C, CD3CN/H2O
Type C peptide mimic : H-H TOCSY
ppm
7.27.47.67.88.08.2 ppm
9
8
7
6
5
4
3
2
1
Phe Hβ
Phe Hβ
Phe Hα
Phe NH
Gly1 Hα
Gly1 Hα
Gly1 NH
Gly2 Hα
Gly2 Hα
Gly2 NH
Ala NH
Ala Hβ
Ala Hα
In collaboration with K Haustraete, K Gheysen, Prof dr J
61/75
62. Type A peptide mimic : H-H TOCSY
H (NH) signals
700 MHz, 25°C, CD3CN/H2O
HO
O
NH
O
H
N
NH
N
H
H
N
N
H
N
H
H
N
O
O
O
O
O
H
N
N
H
O
O
OHO
O
O
In collaboration with K Gheysen, Prof dr J Martins
62/75
64. .
1H-1H nOeSY (RED)
600 ms mixing time,
700 MHz, 25°C, CD3CN/H2O (5/1)
HO
O
NH
O
H
N
NH
N
H
H
N
N
H
N
H
H
N
O
O
O
O
O
H
N
N
H
O
O
OHO
O
O
nOe contact
NH H alpha
N to C direction
ppm
7.27.47.67.88.08.28.48.6 ppm
9
8
7
6
5
4
3
2
1
H alpha
Phe
NH Leu
nOe contact
Type A peptide mimic: H-H NOESY
In collaboration with K Gheysen, Prof dr J Martins
64/75
66. Type A peptide mimic: H/D exchange
BLACK: CH3CN/H2O RED: CH3CN/D2O
In collaboration with K Gheysen, Prof dr J Martins
66/75
DO
O
ND
O
D
N
ND
N
D
D
N
N
D
N
D
D
N
O
O
O
O
O
D
N
N
D
O
O
ODO
O
O
NH Phe
(7.65 ppm)
67. 7.11 ppm
H orto (Phe)
7.18 ppm
H meta (Phe)
1.35 ppm
Hβ (Leu)
nOe
contacts
Type B peptide mimic: H-H NOESY
< 5Ǻ
H alpha NH
N to C direction
NH
NH
HO
O
O N
N
H
N
H
N
O
O H
N
O
OH
ON N
R
O
N
H O
OH
ONN
H
H H nOe cont act
BETWEEN
STRANDS
H
H
H
600 ms mixing time
700 MHz, 25°C
CD3CN/H2O (5/1)
In collaboration with K Gheysen, Prof dr J Martins
67/75
68. Type C peptide mimic: H-H ROESY
ROESY: Off-resonance
H alpha NH
NH H alpha
In collaboration with K Gheysen, Prof dr J Martins
NH
NH
HO
O
O N
N
H
H
N
N
H
H
N
O
O
H
N
O
OH
ON N
NH2
O
O
O
O OH
nOe contact
non adjacent
residues
(1-8)
H
H N
H
N
O
O
N
NN
OH
O
TURN-like-STRUCTURE
4.97 ppm
triazole-CH2-CO
7.54 ppm
NH Gly2
4.97 pp
“Hα” triazole-C
nOe
crosspeak
7.54 ppm
NH Gly2
4.97 pp
“Hα” triazole-C
7.54 ppm
NH Gly2
4.97 pp
“Hα” triazole-C
7.54 ppm
NH Gly2
4.97 pp
“Hα” triazole-C
4.97 pp
“Hα” triazole-C
nOe
crosspeak
7.94 ppm
NH Gly
68/75
72. Dissociation constants (Kd)
In collaboration with V Malanchin, Dr. F Lynen, Prof dr P Sandra
72/75
Strong interactions
range
High affinity ligands
Type A 128 μM
Type B 158 μM
Type C 124 μM
Weak interactions range
Low affinity ligands
(μRL-μR)
Matemathical model
applied
Corresponding equation
Bimolecular binding
curve (model 1) ( )
( )[ ]
[ ]LK
KL
d
dARL
RRL 1
1
max,
1 −
−
+
−
=−
µµ
µµ
Double reciprocal
(model 2)
( ) ( ) [ ] (bRRLRRL LK µµµµµ
+
−
=
− max,
111
X- reciprocal
(model 3)
( )
[ ]
( ) ( RbRRLb
RRL
KK
L
µµµ
µµ
+−−=
−
73. O
O
NO2
NHFmoc
HO
O
NO2
NHFmoc
Wang
Resin
V.1IV.1
Wang resin
MSNT, MeIm
DMF/ CH2Cl2
NH
NH
HO
O
O N
N
H
N
H
N
O
O
H
N
O
OH
ON N R1
R2
O
N
H
R3
O
OH
ONN R4
Type B
C-SPPS
Sequental
double click
Type C
C-SPPS
Linear-SPPS and click
NH
NH
HO
O
O N
N
H
H
N
N
H
H
N
O
O
H
N
O
OH
ON N R4
R3
NH2
O
R1 O
R2 O
R5
DO
O
ND
O
D
N
ND
N
D
D
N
N
D
N
D
D
N
O
O
O
O
O
D
N
N
D
O
O
ODO
O
O
General Conclusion
TYPE A MODEL PEPTIDE
SPPS: HBTU/DIPEA LONG REACTION TIMES
NMR: CAVITY LARGER THAN 5 Ǻ
ACE: WEAK INTERACTIONS 17β-ESTRADIOL
73/75
76. ACKNOWLEDGEMENTS
Prof. dr. A. Madder
Prof. dr. J. Martins
dr. Frederic Lynen
Prof. dr. F. Du Prez
Prof. dr. P. Sandra
Prof. dr. J. Van der Eycken
dr. Steven Van der Plas
dr. Els Van Hoeck
dr. Talha Gökmen
Sara Figaroli
Vivienne Malanchin
UGent people
Jurgen Caroen
Jan Goeman
dr. An Clemmen
Katelijne Gheysen
Katrien Haustraete
Tom Parveliet
UVA people
Prof. dr. Jose Martin
Marie-Curie Foundation
University of Ghent
77. TWEEZER PEPTIDE MIMICS
OF THE ESTROGEN
RECEPTOR FOR
DETERMINATION OF
ENDOCRINE DISRUPTOR
CHEMICALS
Rosa Romeralo-Tapia
Supervisor: Prof. Dr. Johan Van der Eycken
Laboratory for Organic and Bioorganic SynthesisThesis submitted to obtain the degree of
Doctor in Sciences: Chemistry
13 October 2011
Editor's Notes
CÓMO SE DIFERENCIAN (A PRIORI-ANTES DE NOESY) GLY1 DE GLY2?
Receptores artificilaes, ya con uso de esctructuras no proteicas,
quimica orgaica mas amplia
Uso
Tipo de sintesis
Receptores artificilaes, ya con uso de esctructuras no proteicas,
quimica orgaica mas amplia
Uso
Tipo de sintesis
