Transcript What Physiologists Working in Industry Do

What Physiologists Working in
Industry Do
Physiologists in Industry Committee (PIC)
American Physiological Society
Objective
The purpose of this presentation is to 1) inform you
about the responsibilities and types of work a Physiologist
performs as a member of the pharmaceutical, biotech, or
nutritional industry; and 2) describe some of the primary
personal attributes necessary to succeed in this
environment.
Science and Drug Discovery
• The drug discovery process requires concerted efforts of scientists
across many disciplines (Biochemists, Chemists, Molecular Biologists,
Pharmacologists, Physiologists, Technologists, etc…) to advance a
project from initial scientific principles (an idea or hypothesis) to a
clinical candidate, and ultimately a drug to treat human disease.
• This extremely challenging undertaking is pursued under strict
guidelines regulated by federal (i.e. FDA, USDA) and international
(EMEA) agencies. Successful drug discovery requires critical
thinking, organizational skills, creativity, as well as flexibility and
resourcefulness.
• In short, success in drug discovery requires well-trained, disciplined,
and rigorous scientists. The process is one in which Physiologists
can assume many critical roles. Do you fit these criteria?
Scope of Scientific Activities of Physiologists
in Drug Discovery
Activities are listed from first discovery principles (hypothesis generation and
testing) to clinical trials and submission of an NDA (New Drug Application)
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Target discovery and validation
Proof-of-concept studies
Development of in vitro efficacy models
Mechanism of compound action
Development of in vivo models (normal function and disease)
Pharmacokinetic studies
Pharmacodynamic studies
Assay development
Ex vivo functional studies
Disease efficacy testing
Development and utilization of biomarkers
Safety pharmacology
Interpretation of clinical results
Exploration of additional indications
Regulatory submission for product and claim approval
Description of Bench Science Activities of
Physiologists in Industry (Part I)
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Target Discovery and Validation: Studies of disease mechanisms
using molecular and genomic approaches including genetic
association studies in humans, knockout and transgenic animals,
engineered tool compounds to identify or confirm a biochemical
target
Proof-of-Concept Studies: Studies are conducted to address the
questions: does the enzyme, receptor, channel, etc… play a role in
the physiological or disease process? Is the target of interest be
activated/inactivated in this process?
In Vitro Efficacy Testing: Develop and use assays to test the
hypothesis in simple systems such as isolated proteins and/or
cell-based systems. These assays may often use high-throughput
or multiplexed (multiple readouts) platforms.
Examples of Bench Science Activities of
Physiologists in Industry (Part II)
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Mechanism of Compound Action: Studies are designed to address
questions such as does the compound affect enzyme, ion channel,
or receptor activity? Does the compound affect a specific signaling
pathway? Does the compound affect genetic regulation (expression)
of the target?
Development of Disease Models: Studies are designed to address
questions such as: does the cell, tissue, or animal model resemble
the human condition? Is the model valid? (i.e. does the
pathophysiology respond to current pharmacotherapies?)
Examples of Bench Science Activities of
Physiologists in Industry (Part III)
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Pharmacokinetic (PK) Studies: generally considered in terms of “what
the organism does to drug” and studies are designed to address
questions such as: what is the maximal plasma concentration of drug
after dosing and when does this occur? how much drug gets to the
tissue of interest and how widely is the drug distributed in the body?
how quickly is the drug cleared after dosing?
Pharmacodynamic (PD) Studies: generally considered in terms of
“what the compound does to the organism” and studies are designed
to address questions such as: is the biochemical target affected by the
compound and to what extent? Is the tissue of interest affected?
Examples of Bench Science Activities of
Physiologists in Industry (Part IV)
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Ex vivo functional studies: Evaluation of integrated tissue or whole
organ function with ability to carefully control dose, duration, and
exposure to compound (e.g. isolated working heart, isolated perfused
kidney, isolated blood vessel, brain slice, etc…)
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Disease efficacy testing: Acute and/or chronic studies are conducted
to address questions such as: does the compound modify disease
progression in vivo (i.e. prevention model)? Can the compound
regress the disease (treatment model)? How does the efficacy of
the compound compare to existing pharmacotherapies or potential
co-therapies?
Development and utilization of biomarkers: Studies are conducted to
investigate whether there a quantifiable blood or urinary biochemical
marker that predicts severity or progression of the disease? Can the
marker serve as a surrogate for long-term efficacy of the compound?
Examples of Bench Science Activities of
Physiologists in Industry (pt V)
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Exploration of additional indications: Evaluate experimental results
and literature to determine whether additional scientific opportunities
exist (i.e. does the mechanism of action or signaling pathway play a
role in other conditions other than the primary indication?)
Interpretation of clinical results: mechanistic and/or theoretical
evaluation of unexpected clinical events (utilization and identification of
appropriate biomarkers to track mechanistic or pathophysiological
responses to agent).
Preclinical safety pharmacology: Studies are designed to address
questions such as: what is the therapeutic index of the compound?
What is the incidence of adverse events in major organ systems (e.g.
cardiovascular, renal, gastrointestinal, respiratory, CNS) at multiples
(3x, 10x, 30x) of therapeutic plasma concentrations of the compound?
Non-Bench Science Activities of
Physiologists in Industry
Industry scientists have many responsibilities beyond benchwork…
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Training and supervision of technical staff
Generation of novel scientific and technical hypotheses
Rigorous design, analysis, and interpretation of experiments
Presentation of scientific concepts and business applications of
research to various professionals to enable business and scientific
decisions
Participation in project team and strategic planning meetings
Writing of technical reports and scientific manuscripts
Documentation of all scientific observations and submitting patents
Planning facility development and resource (people, space, equipment)
deployment
Participation in the preparation and submission of documents (IND:
Investigational New Drug; NDA: New Drug Application) to the Food &
Drug Administration (FDA)
Personal Attributes of Successful Industry Scientists
(Part I)
To be successful in discovering and developing a new drug, you must
participate in a process that requires concerted efforts by many people
across many departments and disciplines. Specific attributes are required
to succeed in this fast-paced environment…
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Critical Thinking: Industry scientists identify key issues and deliver
timely scientific responses to discovery projects and business
development opportunities
Team and Collaborative Behavior: Industry scientists network with
internal/external scientists to assure access to current and innovative
technologies and scientific advances.
Interpersonal Skills: The drug discovery and development process is a
huge undertaking, successful industry scientists foster a cooperative
spirit, and work well with other scientific, regulatory, clinical, and
business professionals.
Personal Attributes of Successful Industry Scientists
(Part II)
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Strong Communication Skills: Effective communication of ideas
whether one on one, in small groups, or through formal presentations.
Writing is clear, well-organized and logically developed; the audience
is taken into consideration.
Efficiency: The demands on an industry scientist are many (various
experimental models to run and multiple compounds to test). Thus,
industry scientists are constantly challenged to implement new
processes
to streamline procedures while maintaining rigorous standards.
Flexibility: Effectively initiates change as needed and adapts to
necessary changes in operations or strategies; also initiates new ways
of accomplishing work.
Leadership: Industry scientists drive operational plans and develop
and implement tactics to deliver results by set timelines. Leads by
appropriate actions and behaviors; inspires and guides others to
achieve corporate and personal goals.
Physiologists in Industry:
Target Discovery and Validation
Target discovery studies investigate disease mechanisms using molecular and genomic
approaches in knockout and transgenic animals. Physiologists use these approaches to identify
or confirm a role for biochemical targets in organ and organism function and dysfunction.
Below: gene knockout of PKC improves cardiac performance following pressure overload
(TAC), while transgenic overexpression of PKC impairs cardiac performance.
PKC knockout (Prkca -/-) mice have
enhanced cardiac ventricular performance
Overexpression of PKC (Prkca) in transgenic mice
reduces cardiac ventricular performance
Reprinted with permission from Braz et al. Nature Med 10(3), 2004
Physiologists in Industry:
Proof-of Concept Studies
Proof-of-concept studies address whether a biochemical target plays a role in a disease process
(i.e. is the target of interest activated, inhibited or differentially expressed in disease?)
Physiologists use various approaches to learn whether a biochemical target is involved in disease.
Lower left panel: over-expression of the cardiac enzyme, calcineurin (CN) transgenic mouse,
induces cardiac hypertrophy; lower right panel: aortic-banding increases cardiac CN expression
(A) and activity (B), treatment with CsA, a CN, inhibitor prevents aortic-banding induced cardiac
hypertrophy (C).
C
Reprinted with permission from Molkentin Circ Res 87, 2000
Reprinted with permission from Lim et al. Circulation 101, 2000
Physiologists in Industry:
In Vitro Efficacy Testing
In vitro efficacy studies employ assays utilizing simple systems (e.g. isolated proteins and/or
cell-based systems). These preparations allow Physiologists to carefully control experimental
conditions and compound concentrations while measuring and comparing responses and
behaviors of large numbers of compounds. Below: a classical competition curve utilizes
radioligand binding techniques to evaluate the ability of compound Y to compete for receptor
subtype binding, in turn generating affinity and potency data.
[H3] Ligand Bound (% control)
Compound Y Competition vs. Receptor Subtypes 1 and 2
100
75
50
IC50 (nM)
25
0
-13
subtype 1
0.24  0.04
subtype 2
0.30  0.04
Cmpnd Y vs 1
170  92
Cmpnd Y vs 2
40  21
-12
-11
-10
-9
-8
[Compound], M
-7
-6
-5
Physiologists in Industry:
Mechanism(s) of Compound Action
100
response to Angiotensin I bolus
% inhibition of the mean arterial pressure
Enalapril blocks the blood pressure response to Angiotensin I: The
mechanism of action is the antagonism of angiotensin converting enzyme
80
60
40
20
0
-120
-60
Enalapril
(10 mg/kg)
0
60
Angiotensin I
(300 ng/kg, IV)
120
180
Time (min)
240
300
360
Physiologists in Industry:
Development of Disease Models
Ultimately, drugs developed through the discovery process must be able to modify
disease progression and outcome. Physiologists develop models of disease for drug
discovery, and must have an thorough understanding of normal and pathologic ranges
of functional parameters. A valid disease model must involve many of the critical
biochemical pathways and display many clinical findings of the human disease state.
Below: Surgical instrumentation and induction of chronic pacing-induced heart failure.
Surgical Instrumentation
Pacing-Induced HF and Recovery
1.
2.
3.
Cardiac function and coronary flow are
measured in the conscious state by chronic
instrumentation.
Heart failure (elevated end diastolic
pressure, reduced ejection fraction and
cardiac reserve) is induced by right
ventricular pacing for 3-4 weeks.
Recovery from heart failure is allowed by
the termination of pacing for 5-6 weeks
after developed heart failure.
Physiologists in Industry:
Pharmacokinetic Studies
Pharmacokinetic studies characterize how the body handles a compound. Physiologists
work alongside Medicinal and Bioanalytical Chemists to determine if a compound is orally
bioavailable and will achieve adequate plasma and tissue exposure and duration prior to
undertaking an efficacy or chronic disease modification study. Below: Oral administration of
Compound X (10 mpk) exhibited good fractional bioavailability (F% = 54%) and plasma halflife (t1/2 = 6 hours).
Pharmacokinetic Characterization of
Compound X
Plasma Concentration (pg/ml)
1500
iv dose (10 mpk)
oral dose (10 mpk)
1250
i.v. dose Cmax = 1004 pg/ml
1000
oral dose Cmax = 535 pg/ml; Tmax = 3 hrs, F = 53%
750
500
t1/2 = ~6 hrs
250
0
0
4
8
12
16
Time (hrs. post-dosing)
20
24
Physiologists in Industry:
Pharmacodynamic Studies
Pharmacodynamic studies characterize the effects of a compound on the body. Certain
enzymes are activated (e.g. phosphorylated) or modified (e.g. glycosylated) in disease
processes. For example, to determine if a compound will inhibit an enzyme of interest,
Physiologists determine whether enzyme phosphorylation and kinase activity are suitable
pharmacodynamic indices for compound activity. Below: intraperitoneal administration of a
physiological stimulus dose-dependently increased enzyme phosphorylation and kinase
activity, providing a model to assay compound activity against this enzyme’s activity.
Western Blot – Phosphorylated Enzyme
vehicle
3 mpk
Immunoprecipitation Kinase Assay
10 mpk
35000
p-TXXX
*
p < 0.01 vs. vehicle
*
p-TXXX / Total enzyme
0.12
*
*
p < 0.01 vs. vehicle
0.10
*
0.08
0.06
0.04
Counts per Minute
30000
Total
Enzyme
*
25000
20000
15000
10000
5000
0.02
0
Vehicle
0.00
vehicle
3 mpk
10 mpk
3 mpk
10 mpk
Physiologists in Industry:
Ex Vivo Functional Studies
Ex vivo studies evaluate integrated organs and organ systems from normal and diseased
organisms. These preparations allow Physiologists to carefully control experimental
conditions and organ compound exposures while measuring and comparing functional
responses in normal and diseased organs. Below: an isolated working heart preparation
allows careful control of preload, afterload, heart rate, and compound exposure.
Cardiac Systolic Performance Endpoints
LV Systolic Pressure
mmHg
LV End Diastolic Pressure
mmHg
Left Atrial Pressure
mmHg
Aortic Mean Pressure
mmHg
Max +/-dP/dt
mmHg/sec
Peak Pressure
mmHg
Time to Peak Press (TPP)
msec
TPP/PP
msec/mmHg
Diastolic Function Endpoints
tau
msec
1/2 Relaxation Press (1/2 RP)
mmHg
1/2 Relaxation Time (RT1/2)
msec
(RT1/2) / (1/2 RP)
msec/mmHg
Physiologists in Industry:
Disease Efficacy Studies
Drugs developed through the discovery process must show efficacy in modifying disease
progression and outcome. Physiologists develop models of disease for drug discovery, and are
required to have a thorough understanding of normal function as well as the pathology of
disease states. Last, models of human diseases for drug discovery must be amenable to
standard pharmacotherapies. Below: Effects of varying delay (7 day vs. 30 days post-MI) of
ACE inhibitor treatment on a) survival, b) cardiac hemodynamics, and c) morphology after
myocardial infarction (MI).
b
a
untreated
ACEi 30d post-MI
ACEi 7d post-MI
Reprinted with permission from Mulder et al. Circulation 95, 1997
c
Physiologists in Industry:
Development and Utilization of Biomarkers
Blood and/or urinary biochemical markers that are correlated to disease severity allow tracking of
disease progression and regression following drug therapy. Physiologists a) develop disease
models that have biomarker profiles similar to those observed in human disease, b) evaluate
biomarker responses to standard and novel therapeutic agents, and c) develop assays to quantify
biomarkers. Below: Kaplan-Meier survival curve for mortality and morbidity in human heart
failure patients based on plasma brain natriuretic peptide (BNP) concentrations. Thus, plasma
BNP is associated with disease severity and may be used in place of more expensive and timeconsuming assays.
Reprinted with permission from Anand et al. Circulation 107, 2003
Physiologists in Industry:
Preclinical Safety Pharmacology
• Studies can be designed to address specific questions regarding safety
of a compound with respect to a) acute plasma concentrations of drug
(does increasing the plasma concentration of a drug 10-fold above
the therapeutic level induce cardiac electrophysiological abnormalities?)
or b) following chronic dosing. For example:
• Determine the effects of increasing drug concentrations after dosing for
7 or 28 days various organ function, morphology, and histology
• Determine “No Adverse Events Level” (NOEL) for compound
• Federally required data in 2 species before First Time in Human (FIH)
testing