Transcript BioTech at Regeneron: Leveraging Basic Science to Create

Biotech & Pharma
The Science,
The Jobs, & Skills for Success
Neil Stahl Ph.D.
Regeneron Pharmaceuticals
Overview
• Science at a Company vs. Academia
• Attributes for Success at a Company
• Biotech vs. Big Pharma
• Biotech : Innovation and Risk
• Drug Development 101
• Job Opportunities Outside of “Research”
• Getting Hired
My Experience
•
BS Zoology Duke 1978
 Duke Marine Lab 1977, 78, 79 brought me to Science
•
PhD Biochemistry Brandeis 1979-1985
 Quantitative fundamentals of equilibria, kinetics,
& how to make a conclusion
•
Post-Doc at UCSF with Stan Prusiner - Scrapie Prions
 Learned many fields ranging from protein chemistry, cell biology,
transgenics, human genetics
•
Regeneron Discovery, 1991 (65 employees)
 Explored mechanisms of how cytokine receptors are activated and activate
cytoplasmic signaling pathways
 Figured out a way to make tight binding cytokine blockers based on the
mechanism of cytokine activation, using multiple cytokine receptor
components in a recombinant fusion protein = “Cytokine Trap”
•
Regeneron Drug Development (1999-2005; now 565 employees)
 Established preclinical development group - put 3 Traps into clinical trials
 Joined Senior Management - reorganized program management, clinical
project teams, jointly manage all aspects of Research and Clinical
Development, present to investors, analysts, potential partners
Science at a Company
• Scientific endeavor on a project can be carried out at a scale
that is very rare in a University setting
• Teams of competent people aligned toward a common goal can
accomplish more than any individual scientist
• Discoveries can be translated into therapeutic opportunities with
the potential to create new drugs and technologies
 Understand molecular and cellular pathways defining a
particular biology and how it goes wrong in disease
 Create a drug to impact those pathways
 Explore how that drug works in animals and humans
 Design Clinical Program to prove that the drug is safe &
effective
 Register the drug with the FDA and Rest of World
Differences Between Academia & Industry
• You will have access to far more resources, equipment, core
facilities, and collaborative colleagues to advance your project
•
You will be required to work on projects of the company’s
choosing
• You may be asked to switch to (or add on) new projects
• Although you will report to one person, you will interact with
many Scientists instead of a single PI
• Participate and present in cross-functional meetings where data
is vetted and the future directions of a project are established by
discussion and consensus
 More heads are better than 1!
• You are likely to publish and attend scientific conferences
Some Myths of Industry
•
You have failed if you don’t pursue an academic position
 That’s what some told me, but there are many, many incredibly
competent people doing Science & Drug Discovery in Industry
•
The working day is 9-5
 Hard, effective work is expected and rewarded!
•
Compensation is dramatically better than academia
 Entry level scientist positions (3-5 year postdoc) are compensated
similarly to Assistant Professors, but much better than post-docs,
and there are stock options!
 However, opportunity for advancement is more frequent and more
rapid than Academia
•
You never get to publish
 I published more rapidly at Regeneron than anywhere else! Also,
compensation is based on contributions beyond publishing
•
You can’t move from Industry to Academia
 More and more, Universities value Industry experience and
perspective, making a reverse move more likely
Attributes for Success at a Company
• Team player who can collaborate effectively with others
• Ability to become interested in a wide variety of different
scientific areas - learning is a continuous Life-long experience!
• Superb analytical, communication, and presentation skills
• All of us have particular skills that make us good Scientists,
although my exact skill set may not be the same as yours
• Contribute your particular talent and expertise toward the
common goal
• Success means that your project grows so that hundreds of
people work on it!
Biotech
vs
Big Pharma
Often more innovative, high-risk scientific
approaches
Typically more traditional small molecule
Drug Discovery, unless partnered with
Biotech
More informal working environment, with a
“we’re all in this together” spirit.
Typically more hierarchical
A “do what it takes to get the job done”
attitude that may provide more variety
Employees can become pigeon-holed in a
particular function.
Larger organizations usually have more
More likely to participate in decision-making rules!
process
Much larger experience base
More resources than Academia, but often
partners with Pharma for expensive late
stage clinical programs
Can bring huge resources to bear on a
project, although there is always internal
competition for resources
Can be acquired, have layoffs, or slowly go
out of business
Can be acquired, or have periodic layoffs
More opportunities for advancement than
Academia or Pharma if company grows
Base compensation often higher than
Biotech, but usually doesn’t have as large a
stock option upside
Stock options can provide financial windfall
if company successful
Promotion may occur more slowly
Biotech : Innovation & Risk
•
Biotech companies have traditionally been founded to exploit cutting edge ideas
and technology. Examples include:
 Using our own cytokines, growth factors, and enzymes as drugs
 Engineering human fusion proteins, combining functionalities to achieve
new properties
 Creating Humanized and Human Monoclonals as drugs
 Transcriptional control
 siRNA
 Ribozymes
 Aptamers
 Gene Therapy
•
Many Biotech ventures are unsuccessful, often because there is not a realistic
business plan of how to create an income-generating product before their ability
to raise money runs out
•
You need to assess whether the company’s scientific and business plan makes
sense, their history and future potential of raising capital, partnering deals they
have closed, and how soon they will generate revenue
The Promise of the Human Genome Sequence
• The Hype:
 Drug Development will be revolutionized following the
identification of novel genes in “druggable” classes
• The Fact:
 Identifying novel genes is the first baby step.
 Understanding their biology and creating therapeutics
against them is the difficult step that, in many instances, can
take decades
 Accelerating these steps is the key to creating novel
therapeutic opportunities
Target Validation : Velocigene Allows Rapid Creation of Mutant
Mice, and Detailed Visualization of Expression
1 Nature Biotechnology paper described
10% of KO’s ever made!
Huge Opportunities in Protein-Based Therapeutics
• Good Drug Targets are hard to come by
 Many companies make “Me Too” drugs against targets for
which drugs already exist
• Many Interesting Targets are large proteins (eg Cytokines and
Growth Factors) that drive broad biological responses
• These pathways cause disease if inappropriately stimulated
 These targets are usually not amenable to small molecule
approaches
 Current successful approaches include monoclonal
antibodies that block cytokine action, and receptor - Fc
fusion proteins
• As we learn more about Biology, we will uncover an ever
growing number of Targets that will require protein-based
interventional approaches
Protein Therapeutics - Examples
Success Stories
•
Insulin - First administered to humans in 1922
•
Interferons
•
Erythropoietin - 1989
•
Growth Hormone
•
Enbrel - a receptor-Fc fusion protein
•
Antibodies - eg Herceptin, Rituxan, Remicade, Humira,
Avastin
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Less Successful Stories
•
Mouse immunoglobulins - antigenicity
•
Thrombopoietin (Tpo)- efficacy, immunogenicity
•
Lenercept - Receptor-Fc fusion - immunogenicity
•
Leptin - misunderstood mechanism - efficacy
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Protein Therapeutics Strengths & Weaknesses
Strengths
•
high specificity compared to small molecules
•
Little off-target toxicity - less likely to fail in early trials
•
Block Targets not amenable to small molecules - eg growth factors &
receptors
Weaknesses
•
More difficult to manufacture
•
Potential immunogenicity even from fully human proteins
 Low abundance proteins that don’t circulate
 Protein variants (aggregates, oxidation, deamidation) that break
tolerance
•
More difficult to evaluate toxicology
•
injectable
Regeneron Technology: Heteromeric Soluble
Receptors Form Tight-Binding “Traps”
Antibody Structure
Light
Chain
Ra
Heavy
Chain
Rb
Fc
(Drives
Dimerization)
Fc
Cytokine
Kd = 5 nM
Ra
Kd = 10 pM
Rb
In-Line Heteromeric Traps
Ra
Ra
Ra
Rb
Rb
Rb
•
Must Overexpress 2 cDNAs
•
Must Purify Heterodimer away from
Homodimers
•
Waste Cell’s Production Capacity with Unwanted
Homodimers
•
In-Line fusion of 2 receptors without intervening
linkers creates simple homodimer with very high
affinity
Making a Proof of Concept into a Drug
• Make T-shirts
• Do Preclinical Work
• File Investigational New
Drug Application with FDA
• Clinical Trials
• File Biologics License
Application
PreClinical Development Checklist
•
BioMolecular Engineering
•
Cell line Development - FASTR
•
Process Development
•
Formulation
•
Assay Development
•
Pharmacology
•
Pharmacokinetics
•
Toxicology
•
Regulatory - IND = Investigational New Drug Application
BioMolecular Engineering
IL1 Receptor Complex
• Create Trap candidates
with different
receptor order (eg: ab-Fc, ba-Fc,
a-Fc-b, b-Fc-a), different fusion
IL1R Type 1
IL1R-AcP
IL-1
position in receptor sequence +
linkers to increase flexibility
IL1 Single Chain Trap
IL1R-AcP
Extracellular
Domains
• Evaluate for bioactivity, high
Cell Membrane
expression level from CHO cells,
Cytoplasmic
clean folding
Domains
Extracellular
Domains
IL1R Type 1
s s
s s
CH 2
 a-Fc with no extraneous linkers
hIgG Fc
CH 3
Something Old, Something New
Ways to Isolate Over-Expressing Cell Lines
How to isolate clones after transfection:
Traditional:
Random isolation of clones
GOI
FASTR :
Pick
clones
Dilution
clone
ELISA
Isolation based on expression / characteristic of secreted protein
GOI
FACS
ELISA
ELISA
FASTR Cell Line Selection
• Flow cytometry-based Autologous
Secretion Trap
•
CHO Parental cell with doxycylineinducible expression of FcR
•
Binds Trap internally and displays
on cell surface
•
Whole population of transfected
cells can be sorted by FACS with
fluorescent anti-Fc
•
Allows selection of highest
expresser from amongst millions of
transfected cells
•
Turn off expression of FcR after
selection to allow unhindered
secretion for manufacturing
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Process Development
•
Goal is to have protein secreted
from CHO (Chinese Hamster Ovary
cells) which have low viral burden
and make human carbohydrate
structures
•
Batch-Fed Bioreactor Process yield is 1-3 g/L after 10-12 days of
culture
•
Start at 2L scale, eventually to
10,000 L, which yields 10 kg at
expression of 1 g/L
•
3 step purification process (Protein
A, ion exchange, hydrophobic
interaction chromatography) with up
to 70% yield
Formulation
•
Desire high concentration with adequate stability to give > 2 year shelflife
•
Add GRAS (Generally Regarded as Safe) excipients to stabilize protein
from aggregation, deamidation, oxidation, fragmentation
 Polysorbate, sucrose, amino acids, PEG
•
IV formulations generally <10 mg/ml
•
Subcutaneous (SC) - 25-100 mg/ml
•
IL1 Trap: liquid at 50 mg/ml or lyophilized at 80 mg/ml
Assay Development/Pharmacokinetics
• Assays to detect formation of
antibodies against the Trap
• Use to measure PK - how
the blood levels change over
time, which often guides
dosing frequency and active
dose levels
100000
10000
IL -1 Tra p (n g /m L )
• Immunoassays to measure
Trap and their complexes
with target cytokines in
plasma
1000
100
10
Group 4: 3 mg/kg IV
Group 5: 3 mg/kg SC
1
0
0
50
100
150
200
250
Time (hr)
IV & SC pharmacokinetics in Monkeys
•
Injection of bovine collagen II
induces immune response that
results in progressive
autoimmune joint destruction
Injection of zymosan IP at day
30 gives more robust and
synchronous arthritis response
•
Arthritis severity index grades
inflammation, swelling, and
deformity
•
IL1 Trap blocks cartilage
erosion, as well as joint swelling
and deformity
4
Vehicle
3
Trap 10 mg/kg
2
Trap 31 mg/kg
1
0
26
30
34
38
Day
Vehicle
•
CIA model in dba-1 mice is the
most widely accepted model of
rheumatoid arthritis
Trap
•
Arthritis Severity Index
Pharmacology: Murine Model of
Collagen-Induced Arthritis
42
46
50
Toxicology
•
Usually, new drugs are tested at high doses in 2 animal species to identify
NOAEL (No Adverse Event Level) and MTD (Maximum Tolerated Dose)
•
Test drugs at >10x higher doses than expected human dose
•
Many protein therapeutics have strict species specificity, and can only be tested
in primates, but often KO data in animals is predictive of safety issues
•
IL1RI KO shows no adverse phenotype except increased susceptibility to some
types of bacterial infections
•
Moreover, human proteins are often immunogenic in animals
 Immunogenicity in animals not predictive of Ab response in humans
•
IL1 Trap only binds primate IL1
•
6 week toxicology study in monkeys showed no evidence of toxicity, but an
antibody response was observed after a few weeks that resulted in clearance of
Trap from circulation
•
No MTD observed, adequate safety to proceed to clinical trials!
Regulatory
•
FDA regulates testing of
experimental drugs in people
•
Must submit IND - Investigational
New Drug Application
•
Usually takes us ~1 year to
complete, and may involve ~100
people
•
Describes everything you know
about the manufacturing and
structure, PK, pharmacology,
formulation, stability, toxicology,
proposed clinical plan for Phase I
trials
•
FDA gets 30 days to respond,
allowing you to go forward, or
request more information, or to
tweak your clinical trial design…
Clinical Trial Overview
Phase I
•
Safety Dose Escalation in Volunteers or Patients
Phase II
•
Dose Ranging Efficacy Studies to decide on dose and interval
Phase III
•
Proof of Efficacy
•
Treat larger number and broader range of patients to evaluate overall
safety and look for less frequent adverse events (AEs)
 As few as 4 clinical studies (each one a single “experiment”) could
suffice to get a drug approved for use in humans!!
Entry Level Positions in Biotech
Research Post-Doctoral Scientist
• Analogous to Academia, except more resources and mentoring
available
• As in academic post-doc, a good publication record should allow
return to Assistant Professor route
Pharmaceutical Post-Doctoral Scientist
• Contribute to Clinical Development Projects or Core
Technologies in ways that may not result in high profile
publications
• Would lead to a career in Biotech/Pharma
Scientist
• 2-5 years post-doctoral experience
Staff Scientist
• 3 years experience following Post-Doc
Career Opportunities Outside of “Research”
•
Preclinical Development
 Immunoassays & Sample Analysis from Human Clinical Trials
 Formulation Development
 Pharmacology - Assessing Drugs in Animal Models
•
Protein Sciences
 Cell line generation to overexpress recombinant proteins
 Protein characterization
 New technology and assay development
 Protein Manufacturing Process Development
•
Program Coordination & Management
•
Core Facilities
 Methodology Oriented (DNA, in situ, FACS, Mass Spec, Biacore)
•
Clinical
•
Regulatory - understand FDA Guidance, liason for company to FDA, EU
•
Scientific Writing
•
Quality Control
•
Business Development
Getting Hired
Application & Hiring Process
• Typically, job descriptions are posted, applications
solicited
• Human Resource personnel (non-scientists) review
applications, winnowing down to those that match job
description, and pass on to Hiring Scientists
• Unsolicited applications to HR and Hiring Scientists
can sometimes hit paydirt and find an opening before
it’s even listed
CV & Cover Letter Essentials
•
Must communicate to multiple audiences
 Scientists - trying to figure out if you have the raw materials that
they can mold into a productive scientist and useful contributor
 Human Resources - non-scientists checking for a match between
your CV and a job description
•
Usually your First & Only Chance to make a positive impression
•
Should convey your
 Intelligence & ability to communicate (Clear Writing = Clear Mind!)
 Perspective of your field beyond your own project
 Accomplishments - aimed at a non-expert and placed in context of
the open questions in your field
 Skill set - techniques that you really know as well as those for which
you may have a passing knowledge and vocabulary
 Enthusiasm!
CV
•
Same CV can be used for all applications
•
Need not be 1 page - can be 3-4 or longer
•
Research summary
 explain in 1 paragraph your projects and conclusions
 aimed at someone who is not in your field
 Can also briefly describe rotation & graduate research
•
Clearly identify core skill sets
 Don’t exaggerate - you’ll get busted
 just because you have seen a mass spectrometer doesn’t mean
you should list it as a core competency!!!
•
Presentations
•
Awards/Grants
•
Initiatives that you’ve undertaken outside your core requirements
•
Publications - including submitted / in preparation
•
Supervisory & Collaborative experiences
Summary & Technical Skills
Cover Letter
• Ideally should be customized for each application
• Should connect your skill set and experience to the job you are
applying for so that it’s easy for HR to understand and pass on
to hiring scientist
• Should describe your project and findings in the broad context of
your field - often the best way to convey to the Hiring Scientist
that you were not just a skilled set of hands directed by your PI
• Rarely is an applicant “perfect” for the job - often we look for
someone that appears to be smart, communicates well, and can
grow into a job
• Therefore, it’s usually a stretch to say that you can “make
Regeneron a success…”
• More reasonable to emphasize your flexibility and ability to learn
quickly…