TARGET VALIDATION Target validation: Bottleneck

Blue – fluid screening, to do discovery

program, can buy compound lively

How do you identify a target? Target identification – considered fairly

easy, non-problematic for target validation
- Target: the naturally existing cellular or
molecular structure involve in the Due to its non-problematic and easy it is
disease that the drug-in-development is hard “to pick a good idea” – difficult
meant to act on decision to make and evaluate
- Microscopic molecular level. What do Success rate in the Clinic
you want your compound that
physically finds you and have an effect
- so you need to:
o understand the molecules
mechanism of the disease of
interest
o identify a therapeutic target in
this mechanism (Enzyme, gene,
receptor, channel, etc.)
- the more precise you get in that
process or pathway identify exactly
what your target is (so If you protein
you want, your group to bind to or
inhibit/activate, important that the Success rate in the clinic, a different stage
protein would that be will review) the of discovering development when you go
better your off with subsequent later preclinical to phase 1
stages objectives start discovering
developments - 64% of compounds make it to humans
- so understand the basic research in - 44% make it to the phase 2 – evaluate
pathophysiologic in exquisite detail the drug if it will treat a disease – 5-1
- comes out of basic research will make it possible

Prepared by: Fringle

Typical Large Pharma Numbers
• Start 100 new screening programs/assays
¤ New targets ¨
• About 50 find acceptable leads or
chemotypes to pursue ¨
• 20-25 advance into late stage lead
optimization ¨
• 10 programs advance into phase I ¨
Attrition: action or process of gradually reducing
the strength or effectiveness of someone or
something through sustained attack or pressure.
(google)
Why the attrition?
Why Compounds Fail
• Pharmacokinetics
→ ¤ Human ADME
Uses vitro models/testing to test the
pharmacokinetic of the drug
→ ¤ Rodent vs dog vs monkey vs human
Compare the behavior of drug in different
species. Which one resembles the human the
best in terms of pharmacokinetic
• Tox not predicted by animal studies
• Commercial reasons
(Example: Pfizer shut down all new generations
drug that they just experimented due to
commercial issues)
• Lack of efficacy
Most common reasons why compound fails
→ ¤ Biological rationale is incorrect
→ ¤ Relevance of animal models
o Oncology
o Alzheimer’s
European Drug Target Review 2014
Three (3) pillars that will increase the likelihood
of candidate in Phase II trials:
• Deep understanding of the drug
exposure at the site of action
• Target binding of the drug
• Clear expression of functional
pharmacological activity
An in-dept biological understanding of a
molecular target as one of the very early steps
in the entire drug discovery and development
process which can determine later success or
failure of the emerging drug candidate is
required
Lesson learned from the fate of AstraZeneca
Drug pipeline: a Five (5) dimensional
framework
The 5 R’s
• Right target
• Right Patient
• Right Tissue
• Right Safety
• Right Commercial Potential – what’s the
added value beyond what already in the
market; know also the competitive edge
Prepared by: Fringle
Common Rationale Academics
• Target in the literature for 15 years
• Multiple publications: looks like there is
validation
• No one has found a drug…
• start a program!
Things to thing about:
❖ What type of modulator do you need?
➢ Agonist? Antagonist? Partial agonist?
Agonist – drug that occupy receptors and
activate them
Antagonist – drugs that occupy receptors but
do not activate the. Blocks receptor activation
by agonist
To know your target, you must know:
• What type of modulator do you need?
¤ Agonist? Antagonist? Partial agonist?
• Where is the target?
• How will you screen? What is your readout?
Direct measure of target activity or proxy
• What are your secondary assays?
Primary assays use robots for the screen,
assay that how to validate the results came
from the robot
-predict orthology
• Are there species differences?
-test to the homology of different species
• What animal models are available?
¤ Are there standards/known
compounds as a control?
¤ Predictability?
¤ Biomarker?
How many targets are there?
Example of Targets
• Receptors and enzymes
Either intracellular or on membrane surface
¤ Examples: GPCRs, kinases, proteases,
phosphatases
• Transcription factors
• Nuclear receptors
• Ion channels, transporters
• Protein-protein interactions
• Phenotypic screens can be difficult: what is
the target?
Sometimes people propose phenotypic
screens
Phenotypic screens: easy to set up but on the
back end it is quite troublesome
Prepared by: Fringle
What do current drug target?
GPCR’s (Rhodopsin-like) – Majority
Where Do Targets Come from?
• Historically: natural products
¤ Herbal medicine, snake venom –
result physiological response
• Clinical observations
Collaboration with clinicians
¤ Side-effects known drugs: could be
useful in other indication
• Rational approaches based on
biochemistry/biology
• Understanding genetic mutations in people
• Screening/systems biology
-Omics
• New platform technologies
¤ Genomics, proteomics,
pharmacogenomics
¤ RNAi screening
¤ Pathway analysis
¤ Transgenic animals
All these technologies may give hints to pursue
targets
¨ Key question always remains: how to validate?
Validated Target Examples (?)
• Genetic mutations associated with disease
This would give confidence
¤ Alzheimer’s: amyloid precursor
protein (APP) (genetic mutation) and
secretase (enzymes put some APP in
results)
Phase 3 is expensive trials in Alzheimer
¤ HER2 in breast cancer
• Kinases
-Oncology area
• Upregulation in disease process
¤ Inflammation and COX2
Sometimes in disease processes you know
certain enzyme are activated
COX2: Validated?
• Classical COX inhibitors (NSAIDs): not
selective: side effects
¤ Irritation GI mucosa based on
mechanism, drug properties
• ‘90s: discovery of isoforms
¤ COX1: present in most tissues, incluse
GI tract, protective
¤ COX2: upregulated in inflammation
¤ Concept: inhibit COX2 but not COX2
• COX2 inhibitors: Celecoxib (Celebrex,
Pfizer), Rofecoxib (Vioxx, Merck)
¤ Rofecoxib voluntarily withdrawn
2004: increased chance of heart attack
¤ 2005: advisory panels: Merck did not
put it back on market
¤ Now: possible return? Tremeau
Pharmaceuticals: joint pain in
hemophilia; single phase III study
Prepared by: Fringle
Forbes Magazine 2016

When do you consider a target validated?

• Mechanistic studies?
• Analysis in cell culture?
• Analysis in animal models?
• Clinical data (protein or gene expression)?
• Therapeutic intervention: phase II studies?
(shows efficacy)
• FDA approval?

Expert Musing

Target Evaluation Criteria

• Druggable?
→ What evidence is there to support a
drug discovery program?
ACS Med Chem Letters 2015 → Are there small molecule inhibitors
for this class of targets?
→ Availability and predictive value of
animal models?
→ Are there species differences?
→ Biomarker available?
• Structural Information?
The more information the better
Excerpts and Blog Commentary ¤ Protein crystal structure, NMR
• IP (Intellectual Property) Position
¤ Competition?

Prepared by: Fringle

ADDA examples
• Phenotypic screens
→ ¤ HO-1, 14-3-3 theta, TXNIP
→ ¤ What is the target?
→ ¤ Will not be accepted in the ADDA
program anymore
• Defined target examples
→ ¤ CD38
→ ¤ DPY30
→ ¤ LRRK2
→ ¤ Tau-Fyn (protein-protein interaction
project)

Prepared by: Fringle