BioTech at Regeneron: Leveraging Basic Science to Create ... - UMBC

mutebabiesBiotechnology

Dec 6, 2012 (4 years and 10 months ago)

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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.

A “do what it takes to get the job done”
attitude that may provide more variety

More likely to participate in decision
-
making
process

Typically more hierarchical

Employees can become pigeon
-
holed in a
particular function.

Larger organizations usually have more
rules!

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

Stock options can provide financial windfall
if company successful

Base compensation often higher than
Biotech, but usually doesn’t have as large a
stock option upside

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

Less Successful Stories


Mouse immunoglobulins
-

antigenicity


Thrombopoietin (Tpo)
-

efficacy, immunogenicity


Lenercept
-

Receptor
-
Fc fusion
-

immunogenicity


Leptin
-

misunderstood mechanism
-

efficacy






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”

R
a

R
b

Cytokine

Kd = 5 nM

Kd = 10 pM

R
b

R
a

Fc

Light
Chain

Heavy
Chain

Fc

(Drives
Dimerization)

Antibody Structure

In
-
Line Heteromeric Traps


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


Create Trap candidates with different
receptor order (eg:
ab
-
Fc,
ba
-
Fc,
a
-
Fc
-
b
,
b
-
Fc
-
a
), different fusion
position in receptor sequence
+

linkers to increase flexibility


Evaluate for bioactivity, high
expression level from CHO cells,
clean folding


a
-
Fc with no extraneous linkers


Something Old, Something New

Ways to Isolate Over
-
Expressing Cell Lines

How to isolate clones after transfection:

FACS

GOI

ELISA

FASTR
:
Isolation based on expression / characteristic of secreted protein

Dilution

clone

ELISA

Pick

clones

GOI

ELISA

Traditional:
Random isolation of clones

FASTR Cell Line Selection


F
low cytometry
-
based
A
utologous
S
ecretion
Tr
ap


CHO Parental cell with doxycyline
-
inducible 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

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 shelf
-
life


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


Immunoassays to measure
Trap and their complexes
with target cytokines in
plasma


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

IV & SC pharmacokinetics in Monkeys

Pharmacology: Murine Model of

Collagen
-
Induced Arthritis


CIA model in dba
-
1 mice is the
most widely accepted model of
rheumatoid arthritis


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

Trap 10 mg/kg

Trap 31 mg/kg

Vehicle

Arthritis Severity Index

Trap

Vehicle

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…