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C. Frank Bennett Ph.D. Senior Vice President of Research
ISIS-DMPKRx: An Antisense Drug for The Treatment of Myotonic Dystrophy Type 1
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n Stages of Drug Development
n Drug Development Strategies for Myotonic Dystrophy
n Introduction to Antisense Technology
n ISIS-DMPKRx: ¤ Why we are excited about the drug ¤ Where we are in the development process
Agenda
2
Advances in Technology_SCrooke_070212 Isis Pharmaceuticals │ Confidential
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The Stages of Drug Development
ISIS-DMPKRx
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n Identify the therapeutic target → The Gene
n Determine the therapeutic strategy ¤ What to target? ¤ How to target? ¤ When to target?
n Screen to identify the drug candidate ¤ Activity ¤ Safety
Drug Discovery Steps
4
Drug Development
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Federal, Public & Company Roles in Research & Development
Clinical Research
Basic Research
Translational Research
Clinical Research
Basic Research
Translational Research
Government (e.g. NIH) and Public (e.g. MDF) Supported Research
Government, Foundation Supported, Pharmaceutical Industry Research are Complementary
Pharmaceutical Research
There is an ecosystem of science and biotechnology. Public organizations, patient organizations, universities, Congress, FDA, all of this is an ecosystem that is envied in the rest of the world.
– E. Zerhouni,
Director of NIH
“
“5
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1st Clue → What to Target? DM is Caused by a 3 Nucleotide Expansion in the DMPK Gene
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2nd Clue → Nucleotide Expansion Sequesters Proteins A Gain of Function Toxic RNA
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6
Chloride ChannelInsulin ReceptorCardiac Troponin TTau ProteinMyotubularin???
MyotoniaInsulin Insensitivity?Cardiac Arrhythmia?CNS Effects?Myopathic EffectsCataractsTesticular FailureHypogammaglobulinemia
MBNL
CUG-BP
DNA
pre-mRNA
mRNA
DMWD DMPK SIX5
RIBONUCLEAR FOCI RNA BINDING PROTEINDYSREGULATION
ABERRANT mRNASPLICING
DM1 CLINICALFEATURE
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How to Target? Antisense Drugs
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Gene mRNA
DISEASE Transcription Translation
Disease-Causing Protein
How Genetic Information Flows From in DNA to Protein The “Central Dogma” of Molecular Biology
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Antisense Drugs Target RNA, not Proteins
Gene (DNA) mRNA
Transcription
↓ DISEASE
Antisense Drug Inhibition of RNA function
X X
Antisense Drugs can also inhibit the function of a disease causing RNA that doesn’t make a protein (a non-coding RNA) This is nearly intractable for other platforms
Translation
Disease-Causing Protein
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Antisense Drug RNA Target
The Antisense Drug-Receptor Interaction
p ~16-20 base pairs required for specificity
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Antisense Drugs Chemical Modifications Produce Desired Effects in the Body
12 12
Commonly Used Chemical Modifications for Antisense Drugs
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ISIS-DMPKRx: Promotes Degradation of the Toxic DMPK RNA An RNase H Mechanism of Action
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13 13
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Quad
Gastro
c TA0
2
4
6
PBS190403445236445238
HS
A-LR
mR
NA
Leve
l(N
orm
aliz
ed to
18S
RN
A)
Wheeler et al., Nature 488: 111, 2012
Serca-1
PBS 190403 445238 445236
wild
-type
neg.
con
trol
pos.
con
trol
TA
Quad
Gastroc
+ ex22- ex22
+ ex22- ex22
+ ex22- ex22
Proof of Concept Studies: Systemic Delivery of Antisense Drug Reverses Myotonia and Spliceopathy in a Mouse Model of DM1 (HSA-LR)
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Next Step: Demonstrate Reduction of DMPK RNA in Muscle of Different Species
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Proof of Concept: Marked Reduction of Endogenous mDMPK RNA Levels in Mice by an RNase H Antisense Drug
16
0
20
40
60
80
100
120
PBS 486178 (25 mpk/wk)
486178 (50 mpk/wk)
486178 (100 mpk/wk)
mR
NA
Expr
essi
on (%
PB
S)
Tibialis Anterior
0
20
40
60
80
100
120
PBS 486178 (25 mpk/wk)
486178 (50 mpk/wk)
486178 (100 mpk/wk)
mR
NA
Expr
essi
on (%
PB
S)
Diaphragm
0
20
40
60
80
100
120
PBS 486178 (25 mpk/wk)
486178 (50 mpk/wk)
486178 (100 mpk/wk)
mR
NA
Expr
essi
on (%
PB
S)
Heart
0
20
40
60
80
100
120
140
PBS 486178 (25 mpk/wk)
486178 (50 mpk/wk)
486178 (100 mpk/wk)
mR
NA
Expr
essi
on (%
PB
S)
Gastrocnemius
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0
20
40
60
80
100
120
140
Liver Tibialis Anterior Quadriceps Gastrocnemius Kidney Heart
mRN
A Ex
pres
sion
(%
PBS
)
mRNA expression from several monkey tissues
PBS
ISIS 486178
Dose and duration: 40 mg/kg BW; day 1,3,5,7 and then once weekly for the next 12 weeks (total of 16 doses)
Proof of Concept: Reduction of DMPK RNA Levels in Non-human Primates (Cynomolgus Monkey) by an Antisense Drug
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DMPK Drug
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Proof of Concept: Prolong Duration of Effect with a DMPK Targeting Antisense Drug in Monkey Muscle
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DMPK Drug 7 weeks
PBS DMPK Drug 13 weeks
DMPK Drug 19 weeks
DMPK Drug 26 weeks
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The Screening Process The Discovery of ISIS-DMPKRx
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n Generation 2.5 antisense drug
n Promotes degradation of the mutant DMPK transcript by the RNase H mechanism of action
n Delivered by subcutaneous injection ~once per week
ISIS-DMPKRx
20
Constrained ethyl nucleotide 2’-MOE (methoxyethyl)
Partnered with
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The Stages of Drug Development
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n Characterize potential adverse drug effects ¤ Define potential organ or tissue damage induced by the drug ¤ Define reversibility of toxicity
n Characterize pharmacokinetics ¤ Drug levels in blood
n Characterize beneficial pharmacodynamic effects ¤ Does the drug do anything to the target in the body
n Guide safe use in human clinical studies ¤ Determine safe and reasonable starting does ¤ Provide monitoring guidelines for the clinical study
n Provide sufficient data to conclude that patients are not exposed to unreasonable risks
Preclinical Phase Before Testing in Humans
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n Mouse ¤ 13 Week repeat-dose study with 13-week recovery
n Monkey ¤ 13-week repeat dose study and a 13-week recovery
n Standard genetic toxicology and safety pharmacology studies
n Results from toxicology study support continued development of ISIS-DMPKRx
ISIS-DMPKRx IND Enabling Toxicology Summary
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Study Objective: to assess the safety and tolerability of
ISIS-DMPKRx in healthy volunteers n Blinded, randomized, placebo-controlled single ascending dose
n 4 single dose cohorts (randomized 3 active: 1 placebo);
n Doses: 50 mg, 100 mg, 200 mg, 400 mg
n Single subcutaneous injection on Day 1, and subjects are followed for 28 days
n Primary Endpoints: ¤ Safety and Tolerability ¤ Pharmacokinetic measures (plasma drug levels)
ISIS-DMPKRx Phase 1 CS1 Experimental Design - Single Dose
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n Phase 1 SAD in Healthy Volunteers → completed ¤ No safety or tolerability concerns identified ¤ Up to 400 mg single dose tested
ISIS-DMPKRx Phase 1 CS1 Single Ascending Dose Results
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n Planned to start late this year in the United States
n Drug will be given as a subcutaneous injection
n Primary Goals: ¤ Safety & Tolerability, and PK
n In addition, markers of biological activity and clinical biomarkers will be collected
ISIS-DMPKRx Phase 2 Multiple Dose Study in DM1 Patients
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n Antisense drugs demonstrate selectivity for nuclear retained RNAs
n Systemic delivery of Gen 2.5 ASOs profoundly inhibited mouse DMPK RNA levels in normal mice and monkeys: treatment was well tolerated
n Generation 2.5 antisense drugs have long duration of action in skeletal muscle
n ISIS-DMPKRx Phase 1 single dose study in normal volunteers has been completed
n ISIS-DMPKRx Phase 2 multiple dose study in DM1 patients to start late this year
ISIS-DMPKRx Summary
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Isis Pharmaceuticals Sanjay Pandey
Robert Macleod Kathie Bishop Laury Mignon
Kristina Bowyer (Lemonidis)
Husam Younis Scott Henry Dan Norris Viola Kam
Gene Hung Matt Buck
Acknowledgements 28
Dr. Charles Thornton - Univ. of Rochester
Dr. Thurman Wheeler - Univ. of Rochester/ Mass General
Dr. Richard Moxley - Univ. of Rochester Dr. Jack Puymirat - Laval University