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beTherapy
Enteropeptidase: a novel targets for
obesity & type II diabetes treatment
http://www.obetherapy.com
Strategy for obesity drug development
-CNS (appetite, satiety) -Increasing energy expenditure
-Peripheral target
2
Targets for obesity treatment past and present in R&D
Appetite
NPY
Orexin
PTP-1B
AGRP
MG
CNTF
DPP IV
CB1
(rimonabant)
Satiety
OB
DB
CCK
POMC
MCHR1
Tubby
CART
gherlin
FAT
GLP-1
MC4R
PYY (3-36)
5-HT receptors
Serotonin
(Meridia)
Energy
thermogenesis
Fat and Carb.
metabolisms
UCP1
PPAR-γ
UCP2
SREBP1
UCP3
C/EBPs
PKA-IIb
β-Adrenergic R
α-Adrenergic R
ACC-2
Others
SIRT1
H1 receptors
FATPs
PRMD16
BMP-7
SGLT1
PL (xenical)
SGLT2
MTP
Adiponectin
PTP1B
PTP1B
Glucokinase
Histamin
S6K1
3
Molecules withdrawn from Market due to major side effects:
- Thyroid hormone withdrawn 1920 increasing metabolism
- Dinitrophenol (DNP) withdrawn 1930 UCP
- Dexfenfluramine (Redux) (amphetamine) withdrawn at early 1990
- Fen-phen and dexfenfluramine withdrawn from the market in September 1997
- Ephedra was removed from the US market in 2004 over concerns that it raises
blood pressure and could lead to strokes and death
- Rimonabant (Acomplia) It is cannabinoid (CB1) receptor antagonist that acts
centrally on the brain thus decreasing appetite, withdrawn 2008
-Meridia (reductil): Abbott withdraw its obesity drug from the U.S. 2010
-Mediator Servier lab. withdrawn between 2004-2012 in all EU countries.
ObeTherapy approach
Consist of looking for a genotype associated with
inefficiency in energy absorption.
Can be found in rare individuals with a
“lean phenotype” or “starvation phenotype”.
«non variable phenotype»
Congenital Enteropeptidase deficiency
“starvation phenotype”. Cascade of Biochemical events starting with proenteropeptidase
Proteins
Proenteropeptidase
Enteropeptidase
Pepsin
Trypsinogen
Trypsin
Large peptides
Chymotrypsinogen
Chymotrypsin
Proelastase
Elastase
Procarboxypeptidases A and B
Pancreatic prolipase
Carboxypeptidases A and B
Pancreatic lipase
Small peptides
Aminopeptidases
Dipeptidases
Tripeptidases
Free Amino acids
+
Triglycerides
Congenital enteropeptidase deficiency
starvation Phenotype
The pancreas produces proteases, protein-digesting enzymes
that are activated once they enter the duodenum
Pancreas
Membrane-bound
enteropeptidase
Inactive
trypsinogen
Other inactive
proteases
Figure 41.20
Trypsin
Active proteases
Lumen of duodenum
14
Addition of amino acids to the diet
overcomes the problem of growth
retardation.
Polonovsky Et al 1970 Arch. Franç. Péd.
All patients respond favorably to
Addition of Pancreatic enzymes.
Gastroenterology. 1983 Sep;85(3):727-31.
An Esp Pediatr. 1978 Mar;11(3):219-26. Arch Dis Child. 1975 Apr;50(4):277-82.
Enteropeptidase specificities
Trypsinogen sequence recognized by enteropeptidase
Nt-…..-Asp-Asp-Asp-Asp-Lys-Ile-Val-Gly-Gly-….-Ct
Human enteropeptidase model of the catalytic domain (made by MEDIT)
Virtual screening and docking with
potential compounds
11
Rational Drug Design for Enteropeptidase
Serine protease
Common fold
(catalytic domain)
Specific subsites
Crystallographic data
(bovine origin)
Catalytic mechanism
17
Possible sequences of enteropeptidase inhibitors
Structure-function relationship of human enteropeptidase (Biochemistry (Moscow),
2006, 71(2), 113-119).
Ala-Phe-Arg-Boronic
Ala-Phe-Lys-Boronic
13
Acute Effect of OBE lead on Triglycerides absorption
n=3 each group
Acute experiment with C14 radioactivity
n=3 each group
15
Effect of OBE lead on weight DIO of treated mice
n=10 each group
19
Effect of OBE lead on food intake of mice
n=10
Pharmacokinetic of OBE lead compound in mouse
Area
(relative units)
iv, PO route normalized to 1mg/kg
Time (hour)
IV route: OBE lead compound rapidly distributed and eliminated to an undetectable level with an half-life of 21 minutes
PO route: Negligible absorption with a calculated biovaliability 2-3%.
Ex vitro (in plasma), degradation of OBE lead compound was extremely rapid, generating two major metabolites
under limit of quantification after 2 hours. Those two metabolites were not detectable in vivo.
Measurements were done with LC/MS/MS
In conclusion
Due to negligible absorption; fast elimination and rapid degradation in plasma, systemic
exposition to OBE lead compound is negligible after oral absorption.
Advantage of Enteropeptidase Target:
• Peripheral target (intestine), therefore less side effect as
compared to a systemic target.
• No absorption (PK study), no side effect.
• Linked to lean «starvation» phenotype in humans
• Tissue specific
• Non redundant
22
Scientific Advisory Board
Prof. M. Radman, Hôpital Necker, Paris, France; "Académie des Sciences de l'Institut de France".
Prof. A. Levy, The Weizmann Institute of Science, Rehovot, Israel
Prof. T. Baasov, Chemistry Faculty Technion, Haifa, Israel;
Prof. B. Peault, Institute for Regenerative Medicine, UCLA, USA; specialist in cellular models and
biotechnology, pioneer of Systemix.
Dr Y. Champey, former Senior Vice President and Executive Medical Director at Rhône-Poulenc Rorer.
Prof. D. Bensimon, Ecole Normale Supérieure, Paris, France
Prof. M. Rubinstein, Weizmann Institute, Israel;
Prof. D. Ricquier, PhD, Director of CEREMOD/CNRS Unit 9078, Hôpital Necker-Enfants Malades,
France, member of the "Académie des Sciences de l'Institut de France".
Prof. N. Stern Director, Institute of Endocrinology, Metabolism and Hypertension, Tel AvivSourasky Medical Center, Israel.
Arieh Warshal (Nobel prize laureate 2013 in chemistry),University of Southern California US
17
Publications:
- Jolivet, G, Braud, B., DaSilva, B. Gautier, T. , Lagrost, L., Houdebine, L-M., Harosh I* (2014) Validation of APOBEC1
as a new target for obesity treatment using RNAi transgenesis strategy published 12 Sep 2014 | PLOS ONE 10.1371/
journal.pone.0106655
- Harosh, I (2014) Rare Genetic Diseases with Human Lean and/or Starvation Phenotypes Open New Avenues for the
treatment of Obesity and Type II Diabetes (submitted: Current Pharmaceutical Biotechnology).
- Braud S., Ciufolini M., Harosh I* (2012), Enteropeptidase: A Gene Associated with Starvation Human phenotype a Novel
Target for Obesity Treatment. PLOS ONE 7(11): e49612. doi:10.1371/journal.pone.0049612.
- Braud S., Ciufolini M., Harosh I*. (2010), Energy expenditure genes or energy absorption genes: a new target for the
treatment of obesity and type II diabetes, Future Medicinal Chemistry Volume 2, Number 12, December 2010 , pp.
1777-1783(7).
- Harosh I, Braud S and Ciufolini: (2010) Boropeptide inhibitors of enteropeptidase and their uses in treatment of obesity,
overweight and/or diseases associated with an abnormal fat metabolism. US Patent Application No: 2010/0311,690.
Lean genes allow you
to eat without gaining weight...
Nature did it, ObeTherapy can reproduce it.