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 PharmaOften 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

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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”

R R

CytokineKd = 5 nM Kd = 10 pM

RR

Fc

Light Chain

Heavy Chain

Fc

(Drives Dimerization)

Antibody Structure

In-Line Heteromeric TrapsRRRR RR• 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

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ssss

Original Trap DesignR R

hIgGFcssss

Current T :rap Design Single ChainApproachR

RhIgGFc

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: -Fc, -Fc, -Fc-, -Fc-), different fusion position in receptor sequence + linkers to increase flexibility

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

-Fc with no extraneous linkers

IL1R-AcP

IL1R Type 1

hIgG Fc

s ss s

IL1 Single Chain Trap

Extracellular Domains

Something Old, Something NewWays to Isolate Over-Expressing Cell Lines

How to isolate clones after transfection:

FACSGOIELISA

FASTR : Isolation based on expression / characteristic of secreted protein

DilutioncloneELISA

Pickclones

GOIELISA

Traditional: Random isolation of clones

FASTR Cell Line Selection

• Flow cytometry-based Autologous Secretion Trap

• 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

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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 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

0

1

10

100

1000

10000

100000

0 50 100 150 200 250

Time (hr)

IL-1 Trap (ng/mL)Group 4: 3 mg/kg IV

Group 5: 3 mg/kg SC

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

26 30 34 38 42 46 500

1

2

3

4

Day

Trap 10 mg/kg

Trap 31 mg/kg

Vehicle

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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…