Transcript Cyclophilin

Group Meeting Literature Report
Brief Introduction on
Mechanism of Cyclosporin
Speaker ：Geng Hao
Supervisor：Prof. Yun-Dong Wu
Nov.22th 2013
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Outline
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The Discovery of Cyclosporin A and Cyclophilin
Calcineurin and CsA-CyP complex
The mechanism of immunosuppressant-CsA
Cyclosporin analogs
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The Discovery of Cyclosporin A
Cyclosporin A (often shortened to CsA) was
initially isolated from the fungus Tolypocladium
inflatum (多孔木霉), found in a soil sample
obtained in 1969 from Mountain Hardangervidda,
Norway by Dr. Hans Peter Frey, a Sandoz biologist.
Cyclosporin A
Mountain Hardangervidda(哈当厄高原)
No antibacterial activity. The immunosuppressive
effect of cyclosporin was discovered on 31 January
1972 by employees of Sandoz (now Novartis) in
Basel, Switzerland, in a screening test on immune
suppression designed and implemented by
Hartmann F. Stähelin, M.D.
Tolypocladium inflatum (多孔木霉)
Svarstad, 2000,From Norway to Novartis: Cyclosporin from Tolypocladium inflatum in an open access bioprospecting regime.
Biodiversity and Conservation 9 (11): 1521–1541.
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The Discovery of Cyclosporin A
Most peptides are synthesized
by ribosomes, but cyclosporin is
a cyclic nonribosomal peptide of
11 amino acids.
7 N-Methylations
3 uncommon amino acid
(contains a single D-amino acid)
at 1, 2, 8 position respectively
Very famous immunosuppresant!
The first patient, on 9 March 1980, was a 28-year-old woman. Cyclosporin was
subsequently approved for use in 1983. Since then, it has been used to prevent
and treat graft-versus-host reactions in bone-marrow transplantation and to
prevent rejection of kidney, heart, and liver transplants. And it raised the survival
rate from 50% to 70% after internal organ transplants.
Borel JF, 2002. History of the discovery of cyclosporin and of its early pharmacological development. Wien. Klin.
Wochenschr. 114 (12): 433–7
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The Discovery of Cyclophilin
What’s the target of CsA ?
1984
1991
1993
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The Discovery of Cyclophilin
Cyclophilins(Cyp) are a family of proteins that bind
to cyclosporine. Originally discovered as a specific
ligand of Cyclosporin and named based on this.
CypA is the most famous one which has a beta
barrel structure with two alpha helices and a betasheet.
Cyclophilin is not born to bind to CsA
Cyclophilin A is a cytosolic and abundant protein
(0.1% of total cellular protein).
Possesses a peptidyl prolyl isomerase(PPIase)
activity, which catalyzes the isomerization of
peptide bonds from trans form to cis form at
proline residues and facilitates protein folding.
Has been proposed to act as a chaperone(分
子伴侣) in protein trafficking. Play important
role for some virus replication.(HIV,HCV)
Watashi K, et a1. , 2003, 38:1282-1288
Fischer G, Aumuller T; Rev Physiol Biochem Pharmaeol, 2003, 148: 105—150
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Calcineurin and CsA-CyP complex
What happened after CsA bind to Cyclophilin A ?
1984
2002
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The mechanism of immunosuppressant-CsA
Calcineurin (CN) is a enzyme
involved in the T-cell activation.
Known as calcium-dependent
serine-threonine phosphatase,
Calcineurin activates nuclear
factor of activated T
cell(NFATc), a transcription
factor, by dephosphorylating it.
The activated NFATc is then
translocated into the nucleus,
where it upregulates the
expression of interleukin 2 (IL2), which, in turn, stimulates
the growth and differentiation
of T cell response.
interleukin 2
Majed M. Hamawy, et al .Transplantation Review, 2003, 17, 165-171
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The mechanism of immunosuppressant-CsA
Cyclosporine (CsA) binds to cyclophylin
(CpN), forming a complex. The CsA–CpN
complex binds and blocks the function
of the enzyme calcineurin (CaN), As a
result, CaN fails to dephosphorylate the
nuclear factor of activated T cells (NFATc), and thereby blocks the transport of
NF-ATc to the nucleus and the binding of
NF-ATc to the its partner(NF-ATn). No
complex bind to the promoter of the
interleukin 2 (IL-2) gene and initiate IL-2
production. Consequently, T cells do not
produce IL-2, which is necessary for full
T-cell activation. Therefore the CsA
inhibits T cell activation.
T Cell activation
Expression of IL-2 et al
cell factor
Proliferation and differentiat
ion of T cell
CsA-CypA complex
Steffan Ho, et al. Clinical Immunology and immunopathogy, 1996, 80, 40-45
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The mechanism of immunosuppressant-CsA
Cyclophilin A(CypA)
binding domain
Calcineurin binding domain
Qing Huai, Hengming Ke;PNAS, 2002, 99, 12037-12042
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The mechanism of immunosuppressant-CsA
The CyPA-CsA-CN structure
revealed many hydrogen bond
and hydrophobic interactions
between CypA-CsA complex and
CN. CyPA-CsA interacts with Arg122 at the active site of CN,
implying direct involvement of
CyPA-CsA in the regulation of CN
catalysis.
Interfacial interactions between CN and CsA-CyP. A total of 25 residues of CN are
involved in interaction with CyPA-CsA. Red balls represent residues interacting with
CyPA and green balls represent residues interacting with CsA (green sticks).
Qing Huai, Hengming Ke;PNAS, 2002, 99, 12037-12042
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Crystal structure of Cyclosporin A
1984
HR Loosli, H Kessler, H Oschkina,; HELV CHIM ACTA, 1985, 68, 682-704
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Crystal structure of CsA and stucture when binding with Cyclophilin
Cyclosporin A- Cyclophilin C Complex
Cyclosporin A -Cyclophilin D Complex
2-Val Cyclosporin A -Cyclophilin D Complex
Crystal Structure of human calcineurin complexed with
cyclosporin A and human cyclophilin
(U-13C)Cyclosporin A bound to Cyclophilin
[d-(cholinylester)ser8]-cyclosporin Cyclophilin B Complex
Cyclosporin A -Cyclophilin G Complex
Cyclosporin A- Cyclophilin J Complex
Cyclosporin A - murine cyclophilin C complexed
Overlay of Crystal structure of CsA from
different CsA-CyP complexes
2ESL
2Z6W
1CWF
1MF8
1CYA
1CYN
2WFJ
2OJU
2RMC
Amide nitrogen toward the inside direction
N-methylation toward the outside
Beneficial to pass through the membrane
Conformation of CsA in 1MF8
No intermolecular H-bond
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Calcineurin and CsA-CyP complex
x
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Non-Immunosuppressive Cyclosporin Analogs
Cyclosporin A
NIM-811
DEBIO-025
SCY-635
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Non-Immunosuppressive Cyclosporin Analogs
NIM 811
Inhibition of Human Immunodeficiency
VirusType1 Replication by SDZ NIM 811 , a
Nonimmunosuppressive Cyclosporine Analog.
ANTIMICROBLAL AGENTS AND
CHEMOTHERAPY,Aug. 1994,p. 1763-1772
Debio-025
In vitro blood distribution and protein
binding of a new anti-HIV drug, DEBIO-025.
DRUG METABOLISM REVIEWS,2005, 37,
p.29-29
SCY-635
Preclinical evaluation of SCY-635, a
cyclophilin inhibitor with potent anti-HCV
activity. HEPATOLOGY, 2006,44,p.534-535
Gallay PA, Clin. Liver Dis., 2009
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What are we interested in
• The effect of different substituents on the
conformation of Cyclosporin
• How the conformation affect the binding of
Cyclosporin and Cyclophilin /CsA-CyP-Calcinurin
• Design new modification for better different
kinds of activities.
First step : A good computational method to study it.
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Preview
Cyclophilin
What are the different roles they play.
How CypA bound to HIV-1 Capsid
Anti-HCV Mechanisms
of CypA
Welcome to the next literature report!
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