biology services
TRANSCRIPT
Randy Jones, D.V.M., Ph.D.
Diplomate A.B.V.T. & A.B.T.
Vice President Biology Services
Ricerca Biosciences, LLC
May 23, 2007
Biology Services
Introduction
• Animal models screen drug candidates for potentialtherapeutic efficacy
• Confounded by species of animal, metabolism,pharmacokinetics, organ system anatomy, and physiology
• An initial opportunity to integrate biology and chemistry
• Anti-infective, oncology screening, and anti-inflammationmodels are likely to remain important for development ofdrug candidates for an aging population
• Established disease models require less time anddevelopment expense but may lack specificity
• Brief application of anti-infectivity models, in vivo anti-tumorassays, and anti-inflammatory models will be presented
• Typical biotech customer proceeds cautiously with one or twoprojects and moves to combinations of biology and chemistry
• Need to develop their “one-and-only” lead into an IND
• Cash Flow – “do or die”
• Smaller organizations – fewer layers
Biotech Business Model
Is the Bio-Entrepreneur more successful
than Pharma at drug development?
Overview
• Ethical considerations support the judicious use of animals
prior to first-in-human use
• Drugs are not used to treat normal people
• Drug candidates are tested for toxicity on physiologically normal,
juvenile animals (rodent & non-rodent)
• Pharmacology vs toxicology endpoints
• Mechanism of action – homology, specificity….
Animal models of human disease are used to screendrug candidates for potential therapeutic efficacyfocusing on pharmacology and mechanism of action
Overview (Continued)
The predictive nature of the model and its potential toextrapolate to a human disease is impacted by:
• Species of animal
• Metabolism – constitutive and inducible capacity
• Pharmacokinetics – drug-ability
• Organ system anatomy
• Physiology
Metabolite Profile
%Loss of Parent Compound
0
20
40
60
80
100
120
0 5 10 15 20 25 30 35
Incubation Time (min.)
% L
oss o
f P
are
nt
Dog Cyn Monkey Rh Monkey Human Mouse Rat
%Increase in Metabolite Formation
0
10
20
30
40
50
60
0 5 10 15 20 25 30 35
Time (min.)
% I
nc
rea
se
in
Me
tab
oli
te
Dog Cyn Monkey Rh Monkey Human Mouse Rat
Risk Management
! Cyn vs Rh
! Teenage athlete vs
Geriatric poly-pharmacy
! Therapeutic index
! Clinical Indication
! Bimodal or uniform
pharmacogenomics
! FDA/ICH guidelines
Pharmacokinetics
Rapid In Vivo screening
Pharmacokinetic Parameters
• AUC, volume of distribution,
half-life, Cmax, clearance,
bioavailability
Test Material Requirements• Limited amount
• Radiolabel not necessary
0
2000
4000
6000
8000
10000
0 2 4 6 8 10 12 14 16 18 20 22 24
Time (hr)
ng/
mL IV
oral
Parent (Pro-drug)
0
100
200
300
400
500
600
700
0 2 4 6 8 10 12 14 16 18 20 22 24
Time (hr)
ng/mL
IV
oral
Active Metabolite
Integration of Biology and Chemistry
• Saltability
• Crystallinity
- HS-PLM, XRD, DSC, TGA
• Hygroscopicity
- Hydration states
• Solubility
• Stability
• Polymorphism
• Powder Properties
Why is this a Problem?
• Physical-chemical
properties of each
form are different
• The intermolecular
forces in a solid
contribute to the
properties of the solid
• Solubility
• Dissolution Rate
• Chemical Stability
• Physical Stability
• Processability
• Rate of Elimination
• Bioavailability
Animal Models
In Vivo Efficacy• Anti-infective
• Anti-cancer
• Anti-inflammation
• Others
• Obesity
• Diabetes
• Gene Therapy
Work with Clients to Customize Models
Dedicated BSL-2 Animal Rooms
Animal Models of Infection
(Anti-Infective)
Infectious agent introduced & the ability of the drug candidateto relieve the experimental disease process is evaluated
• Thigh Infection Model – bacterial agents (mouse or rat)
Neutropenic animal, end points and target tissues
Antimicrobial efficacy of the drug candidate – plate count data CFU/gramthigh tissue
Pharmacokinetics
Clinical pathology
• In Vitro Assay Support
• Minimum inhibitory concentration, minimum bactericidal concentration, time-kill kinetic assays
Animal Models of Infection
(Anti-Infective)
An infectious agent is introduced and the ability of
the drug candidate to relieve the experimental
disease process is evaluated
• Mouse Sepsis Model – Staphylococcus aureus (MSSA
and MRSA), S. pneumonia, E. Coli, P aeruginosa,
Candida albicans (anti-fungal)
– End points and target tissues
0
10
20
30
40
50
60
70
80
90
100
-1 5 11 17 23 29 35
Day
% s
urv
ival
Infected control
Vancomycin
REP0897 (solution)
REP0897 (suspension)
REP0318 (solution)
REP0318 (suspension)
---- Infected control
---- Vancomycin
---- TA-1 (solution)
---- TA-1 (suspension)---- TA-2 (solution)
---- TA-2 (suspension)
Oncology Screening Models
(Anti-cancer)
In Vivo Anti-tumor Assays (Xenograft models)
• Severe combined immunodeficient (SCID) mice, single subcutaneousinjection x 7 day for tumor induction followed by drug candidate dosingby applicable route and dose levels x 7 days.
• End points - tumor size, histopathology of the induced lesion, clinicalpathology
• Pharmacokinetics
Currently established tumor models at Ricerca:
Cell Line Species Cancer Type
C-33A human cervical
Ramos human B lymphocyte
PC-3 human prostate
A-549 human lung, non-small cell
HL-60 human leukemia, PML
B16-F0 mouse melanoma
Oncology Screening Models
(Anti-cancer)
In Vitro Assays
• Anti-proliferation
• Acute cytotoxicity – lethality or induction of apoptosis
-20
0
20
40
60
80
100
0.10 1.00 10.00 100.00
Conc (!M)
% In
hib
itio
n
-20
0
20
40
60
80
100
0.10 1.00 10.00 100.00 1000.00
Conc (!M)
% I
nh
ibit
ion
Anti-Inflammation Model
An acute efficacy screening model to evaluate impacton the inflammatory response:
LPS Induction of TNF! Release in Balb-c Mice
• Drug candidate administered orally, intraperitoneal, sub-cutaneously
• Lipopolysaccharide (LPS) dosed IP - optimized to provide maximal release of TNF!
• End points – serum/plasma TNF! by ELISA
– Pharmacokinetic satellite group
– Biomarkers
Effect on LPS Induced TNF! Release in Mice
by Single Oral Dose of Test Article
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
TA
1,1
0
TA
2, 10
TA
3,1
0
TA
4, 10
PC
,10
Test Article dosed and dose administered (mg/kg)
Perc
en
t re
du
cti
on
fro
m L
PS
co
ntr
ol
1 hour
between
oral dose
and LPS
dose
4 Hours
between
oral dose
and LPS
dose
Summary
• Animal models screen drug candidates for potentialtherapeutic efficacy
• Confounded by species of animal, metabolism,pharmacokinetics, organ system anatomy, andphysiology
• An initial opportunity to integrate biology andchemistry
• Anti-infectivity models
• Anti-tumor assays
• Anti-inflammatory models
Thank you!
Ricerca Contacts
Ann L. O’Leary, Ph.D.
Manager, Animal Models/Microbiology
440-357-3561
Prabu Devanesan, Ph.D.
Manager, In Vitro DMPK
440-357-3106
Andrea Hubbell
Scientist, In Vitro DMPK
440-357-3753
Randy Jones, D.V.M., Ph.D.
Diplomate A.B.V.T. & A.B.T.
Vice President Biology Services
Ricerca Biosciences, LLC
February 5, 2007
Biology Services