aposense molecular nanomotors (mnms) · 5 additional continuation-in-part applications (cips) are...
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Aposense Molecular NanoMotors (MNMs)The first platform, energized by the membrane
dipole potential, for systemic delivery of Nucleic Acids
Aposense Ltd. 5-7 Odem St., P.O.Box 7119, Petach Tikva 49170, Israel Tel: 972-73-2397-600 Fax: 972-3-9215714 www.aposense.com
September 2017
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CAUTIONARY NOTE REGARDING FORWARD-LOOKING STATEMENTS
The following is a presentation of our Company’s business and operations. This presentation does not constitute an offer to sell or
the solicitation of an offer to subscribe for or buy or sell any of our securities. The information contained in this presentation does
not constitute a basis for making an investment decision and does not constitute a recommendation or opinion, nor is it a
substitute for the discretion of a potential investor. Readers are urged to carefully review and consider the various disclosures
made throughout our immediate and interim reports as filed with the distribution systems of the Israeli Securities Authority
(https://www.magna.isa.gov.il) and with the Tel Aviv Stock Exchange Ltd. (http://maya.tase.co.il/reports/company), which are
designed to advise interested parties of the risks and factors that may affect our business, financial condition, results of operations
and prospects. Certain information included or incorporated by reference in the following presentation may be deemed to be
“forward-looking statements” within the meaning of the Israeli Securities Law, 5728-1968. Forward-looking statements are often
characterized by the use of forward-looking terminology such as “may,” “will,” “expect,” “anticipate,” “estimate,” “continue,”
“believe,” “should,” “intend,” “project” or other similar words, but are not the only way these statements are identified. The following
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changes in regulatory, legislative and/or international standards environment; (-) our success in implementing our business-
development, sales, marketing and/or manufacturing plans; (-) the level of adoption of our future products by medical practitioners;
(-) the emergence or existence of other products that may make our products obsolete; (-) protection and validity of our patents
and other intellectual property rights; (-) the effect of competition by other companies and technologies; (-) the ability to obtain
reimbursement for our products from government and commercial payers; and (-) our ability to raise sufficient funds for completion
of products research and development and/or our ability to partner with strategic partners and/or investors. You should not rely
upon forward-looking statements as predictions of future events. We cannot guarantee future results, levels of activity,
performance, or achievements reflected in the forward-looking statements. You should not put undue reliance on any forward-
looking statements. Any forward-looking statements in this presentation are made as of the date hereof, and we undertake no
obligation to report events or to report the occurrence of unanticipated events that may lead to the actual events, results,
performance, circumstances or achievements concerning our business and operations being different than as envisaged by such
forward looking statements and/or to publicly update or revise any forward-looking statements, whether as a result of new
information, future events or otherwise, except as required by law. Date of presentation: September 2017.
Milestones in the Evolution of siRNA Therapeutics: From Discovery to the Systemic Delivery Barrier
Discovery of gene silencing through injection of dsRNA to worms1998
Discovery of the RNA Induced Silencing Complex (RISC)2000
Discovery of the Dicer endonuclease2001
The Power of siRNA is demonstrated in numerous applications across
numerous fields of medicine; Huge investments in the field by Pharma
Companies
2001-2006
Nobel Prize to Andrew Z. Fire and Craig C. Mello; "for their discovery of RNA
interference - gene silencing by double-stranded RNA”
2006
The Systemic Delivery Barrier: Large withdrawal from the field: “siRNA is
probably non-druggable”
2006 - 2014
First demonstration of clinical benefit of siRNA targeted to the liver (ATTR
amyloidosis)
2014
Renewed interest in siRNA; Strategic alliances
Only for liver-related indications; or for topical administration
2014 -
• Recognition that siRNA may hold a great therapeutic promise
• Pending on solving The Systemic Delivery Barrier
2014 -
Lack of efficacious and Universal Tools for Systemic Delivery of siRNA; Widely-Recognized as The Major Factor that Holds-Back Clinical Implementation of siRNA
“The most intractable problem with RNAi therapeutics has to do with systemic delivery. Nucleic acids such as RNAi are relatively large and carry a negative charge, preventing absorption through the cell membrane….
Considerable efforts have been expended breaking down the delivery barrier. Yet even the combined resources of both Big Pharma and small biotechs have not been enough to solve the problem, at least not yet”.
Pfizer
siRNA therapeutics; The promise and the unmet need
“The siRNA Technology is quite amazing, to be sure. These small pieces of nucleic acid are capable of knocking down genes with extremely high selectivity”
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siRNA: Molecular weight 14,000 Daltons42 negative charges
Classical drug
Genetic drugAspirin: Molecular weight
180 Daltons
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siRNA: The delivery challenge
Delivery of siRNA across cell membranes into cells requires a very large activation energy
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Apo-Si Molecular Nanomotor PlatformA disruptive technology
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A novel internal
powerhouse
Innovative
Molecular
NanoMotors
Breaking-down the membrane delivery
barrier
• Electric voltage measured between:Membrane / water interface (-) membrane center (+)
• Generated by the ordered carbonyl groups of the phospholipids
• Environment: very hydrophobic
• Dielectric constant: 2-4
• Due to hydrophobic environment, results in an enormously strong intra-membrane electric field of 108-109 V/m !!!
The membrane dipole potential
ΔΨ = the trans-membrane potentialΨs = the surface potential
Ψd = the dipole potentialε = the dielectric constant
8Wang L. Annual Review of Biochemistry 81: 615-635, 2012
A novel internal
powerhouse
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Transmission Unit
Detachment Unit
Electrical Unit
Point for “click attachment” to any Oligonucleotide sequence
(DNA or RNA)
The Apo-Si Molecular NanoMotor (MNM):Optimized molecular structure for utilizing
the membrane dipole potential for drug delivery
• Small-chemical Entity
• Proprietary; All in-house IP
• Non Toxic
• Modular
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Innovative
Molecular
NanoMotors
The MNM attachment module
• Standard building block of any oligonucleotide synthesis • The locations and number of moieties can be tailored per the
specific application
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Detachment Module
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MNM; MW ≈700 Daltons
dsiRNA; MW ≈14,000 Daltons
The dsiRNA-MNM construct
Prolonged residence
in body fluids
Trans-membrane
delivery to reach the
cytoplasm
Membrane
surface
Breaking-down the membrane delivery
barrier
Direct entryEndosome-mediatedentry Flip-flop
Endocytosis; Flip-flop; RNA detachment
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MNM:
• Detachment from RNA
• Excretion: Kidneys / Liver
RNA: Interaction with Dicer & RISC
Gene Silencing
Cytoplasm
Robust trans-membrane delivery of Apo-Si-MNM Conjugates; B16 melanoma cells
X 20
Cy3= Red fluorophore
Control: fluorescently-labeled dsiRNA without MNMs
Fluorescently-labeled dsiRNA, conjugated to MNMs
Control MNM-dsiRNA Conjugate
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1
64
78
90
97
0
10
20
30
40
50
60
70
80
90
100
control 15min 30min 1hour 2hours
% P
osit
ive E
ven
ts,
Cy3
Delivery of dsDNA, Attached to Two Apo-Si-MNMs
3T3 cells; Time course; 40nM; FACS
N=3; n=6
Mean±SE
Uptake of Apo-Si-MNM Conjugate is rapid and efficient; 3T3 Cells
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FACS analysis; Phloretin 750µM 3hr, Apo-Si-MNMs Conjugate 40nM, 2hr
Shutting-down the internal membrane electrical field inhibits delivery of Apo-Si-MNMs Conjugate
Complete inhibition by Phloretin
0%
EGFP fluorescence
Field ONNo MNMs
Apo-Si-MNMs
Conjugate
Field ON+ MNMs
Apo-Si-MNMs Conjugate +
Phloretin
1%
Ap
o-S
i D
elive
ry;
(Cy3
)
EGFP fluorescence
Field OFF+ MNMs
95%
EGFP fluorescence
Ap
o-S
i D
elive
ry;
(Cy3
)Control
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Ap
o-S
i D
elive
ry;
(Cy3
)
Shutting-down the membrane dipole potential inhibits delivery of Apo-Si-MNMs Conjugate; dose-related
N=3; n=6
Mean±SE
FACS analysis; 3T3 cells; Phloretin 3hr, Apo-Si-MNMs Conjugate 40nM 2hr
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Deletion of the Apo-Si-MNM negative pole blocked the delivery of oligonucleotides into 3T3-EGFP cells
40nM, 24hr, FACS Analysis
Increased fluorescence
Apo-Si Uptake; Cy3 fluorescence
Apo-Si-MNMControl;
Apo-Si-MNM; deletion
of negative pole
Control
Ce
ll C
ou
nts
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Robust down-regulation of EGFP expression, induced by Apo-Si-MNM-dsiRNA Conjugates
EGFP-3T3
X20
MNM-dsiRNAConjugate
Control:dsiRNA alone
Control:MNM+DNA
Bright Field
High level of gene expression
High level of gene expression
Robust gene silencing (90%) 3T3 cells
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100
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1512
9
0
20
40
60
80
100
120
Untreated 10 nM 40 nM 150 nM 300 nM
% G
FP
IC50= 4.86 nM
Apo-Si-G dsiRNA, nM
% E
GF
P
0 50 100 150 200 250 3000
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40
60
80
100
Robust down-regulation of EGFP expression, induced by Apo-Si-MNM Conjugates in HeLa cells; ELISA,72 h
N=3; n=9Mean±SE
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MNM-dsiRNA
MNM-dsiRNA; nM
Apo-Si-MNM: Systemic bio-distribution of siRNAupon intravenous administration
SPECT imaging of I125- labeled MNM-dsiRNAConstruct ; 80-90 minutes after iv administration
0.0
0.5
1.0
1.5
2.0
2.5
3.0
Heart Lung Liver Spleen Kidney Small intestine Large intestine Lymph nodes
SU
V
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Apo-Si Molecular Nanomotor Platform;performance profile
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Favorable bio-distribution upon systemic administration in vivo rapid and robust distribution of siRNA into various organs (e.g., liver, heart, lung, spleen, kidneys, lymph nodes)
Gene Silencing
• Robust (>80%)
• Nanomolar concentrations
• Dose-responsive manner
• Silencing of both reporter and disease-related genes
• Cytoplasmatic enrichment mechanism
Delivery: • Energized by the membrane dipole
potential
• Universal
• Nanomolar concentrations
• Efficient, into all treated cells
• Rapid
• Dose-dependent manner
• Safe
• PCT and world-wide patent applications
• Novel Compounds, Conjugates & related formulations
• Mechanism of action
• Methods utilizing said Compounds and Conjugates for delivery of molecules and macromolecules, across biological membranes into cells.
• Methods for utilizing said Compounds and Conjugates, for the treatment of medical disorders
Patent Coverage:
• US Patent Application No. 14/830,799 was allowed on April 11, 2017.
• 5 Provisional Patent application were consolidated into a PCT Application, which is now in the National Phase, world-
wide.
• Corresponding applications are now pending in
Israel, Japan, Europe, Canada, China, India, Australia, Korea, Brazil, Mexico, Russia (filing date: 29-March-2015).
• 5 additional Continuation-In-Part applications (CIPs) are now pending in the United States.
• PCT (international) patent application No. PCT/IL2016/050893 is pending (filed: 17-August-2016) covering the same
aspects of the CIPs.
• A new US-patent, covering additional recent developments in the Project is now being submitted.
Patent Applications and status:
Apo-Si: Intellectual Property
Scientific Leadership
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Professor Roger Kornberg, Scientific President:, Nobel Prize Laureate in Chemistry for the year 2006, for his research and discoveries of the molecular basis of eukaryotic transcription (synthesis of RNA from a DNA template).
Professor Ilan Ziv, MD, CEO & CSO. Ilan is a co-founder of Aposense, and the Chief Scientist of the Company. He is an associate professor in Neurology at the SacklerSchool of Medicine, Tel-Aviv University. Ilan is the Inventor of the AposenseTechnology.
Dr. Hagit Grimberg, Ph.D, Senior VP for R&D, has been with Aposense for over 16 years. Hagit has an M.Sc. in Biochemistry, and Ph.D in Physiology & Pharmacology. Hagit has strong background in innovation, research and new product development.
Multiple, Parallel Joint ventures with Pharma Companies
Synergy: Apo-Si delivery platform disease-specific genetic drugs
Delivery modules for siRNA, ASO and beyond
Gene delivery beyond the liver
Potential Income: Upfront payments; milestone payments, profit sharing
MNM
Gene
Delivery
Platform
A Huge, Multi-billion $ profit potential
Apo-Si; Business Model as a Platform Technology Company
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The Vision To introduce an industry standard
for the trans-membrane delivery of genetic drugs
• The opportunityBoth the unmet need (delivery) and the source of energy(dipole potential) are co-localized in the cell membrane
• The solution
• Molecular NanoMotors, internally-energized, for efficacious delivery of genetic drugs
• Small-molecule “chip”, modular, versatile and “click-attached” to any Oligonucleotide sequence
Translating genetic therapeutics into a “druggable” reality 25
Thank you!