Table Of ContentDELIVERY
STRATEGIES
for
ANTISENSE
OLIGONUCLEOTIDE
THERAPEUTICS
DELIVERY
STRATEGIES
for
ANTISENSE
OLIGONUCLEOTIDE
THERAPEUTICS
Edited by
Saghir Akhtar
First published 1995 by CRC Press
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Reissued 2018 by CRC Press
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PREFACE
Antisense nucleic acids (oligonucleotides) can act sequence specifically to modulate gene
expression in living cells and thus are being considered for therapeutic application in
diseases where the genetic target and its sequence have been identified. Indeed, the
concept of antisense oligonucleotide therapeutics has rapidly developed from being a
purely cell culture phenomenon to one that is now undergoing clinical trial evaluations
for the treatment of leukemias and viral infections such as HIV, herpes simplex, and
human papilloma virus.
A major issue in the development of antisense nucleic acids as potential therapeutic
agents has been their effective delivery to target cells. The large molecular weights and
often polar nature of oligonucleotides coupled with their sensitivity to nuclease degrada
tion have posed new challenges for biopharmaceutical scientists in developing suitable
drug delivery systems. With contributions from leading experts in the antisense field, it
is the aim of this volume to highlight the major hurdles to effective delivery and then to
discuss the state-of-the-art strategies currently being developed to improve the delivery
and targeting of oligonucleotides to the desired sites of action in the body.
An overview of the concepts involved in antisense oligonucleotide therapeutics (which
includes the antisense, antigene, and ribozyme strategies) can be acquired from the
introductory chapters by A. Gewirtz and colleagues, N. Chaudhary et al., and by D. Elkins
and J. Rossi in Section 1. The in vivo pharmacokinetic behavior of oligonucleotides will
be an important consideration in the design of a suitable formulation or delivery system
and this area is therefore covered in Section 2 with contributions from the groups of L.
Neckers and V. Vlassov.
The important issue of improving the biological stability of nucleic acids with a view
to improving delivery to target sites is covered in Section 3. This section includes a review
by E. Wickstrom on the general strategies for modifying nucleic acid structure to improve
nuclease resistance followed by chapters reviewing important developments in the use of
self-stabilized oligonucleotides (Agrawal et al.), circular oligonucleotides (E. Kool),
peptide nucleic acids (J. Hanvey and L. Babbiss), and stabilized RNA for antisense and
ribozyme applications (F. Eckstein and colleagues).
Section 4 addresses the major issues of improving membrane transport, manipulating
subcellular distribution, and improving delivery by cell-specific targeting of antisense
oligonucleotide therapeutics. This section begins with chapters by A. Krieg and by R.
Juliano’s group reviewing the mechanisms by which oligonucleotide analogs enter living
cells. A general opinion emerging from these (and other) studies is that uptake of
oligonucleotides is reduced by some form of endocytosis with subsequent accumulation
of these macromolecules into acidic endosomal/lysosomal compartments. This is fol
lowed by the eventual escape of oligonucleotides into the cytosol and/or efflux out of the
cell by exocytosis. Strategies which may bypass endocytosis and thus improve the
intracellular bioavailability of oligonucleotides include the use of cationic and other
liposome formulations, and these are covered in chapters by F. Bennet and by A. Thierry
and G.B. Takle. Alternative strategies aimed at enhancing endosomal exit of oligonucle
otides that have entered cells by endocytosis include the use of pH-sensitive liposomes
(C. Malvy and colleagues) and pH-sensitive viral fusion peptides, a strategy being
developed for the delivery of DNA in gene therapy, but which has obvious implications
in oligonucleotide therapy (E. Wagner and colleagues). The final two chapters highlight
the progress being made in achieving cell-specific delivery of oligonucleotides by either
forming antibody-oligonucleotide conjugates (C. Gooden and A. Epenetos) or by conju
gating oligonucleotides to cell-specific ligands to promote receptor-mediated endocytosis
(E. Carmichael et al.).
I hope that this volume, the first to focus solely on the delivery of these interesting
macromolecules, will serve both to provide an overview of the current strategies being
pursued in the effective delivery of antisense oligonucleotide therapeutics, and to stimu
late new contributions to this exciting and rapidly developing field.
Saghir Akhtar
Birmingham, U.K.
CONTRIBUTORS
Sudhir Agrawal, Ph.D. Christine Chavany, Ph.D.
Senior Vice President Visiting Fellow
Department of Discovery Clinical Pharmacology Branch
Hybridon, Inc. NIH
Worcester, Massachusetts Bethesda, Maryland
Henry C. Chiou, Ph.D.
Saghir Akhtar, Ph.D.
Staff Scientist
Lecturer
Department of Gene Therapy
Pharmaceutical and Biological Sciences
The Immune Response Corporation
Aston University
Carlsbad, California
Birmingham, UK
P. Dan Cook
Helle Aurup, Ph.D. ISIS Pharmaceuticals, Inc.
Professor Carlsbad, California
Max-Planck-Institute Fur
Patrick Couvreur, Ph.D.
Experimental Medizin
Full Professor
Gottingen, Germany
URA CNRS 1218
University of Paris XI
Anna Avroutskaya
Chatenay-Malabry, France
Research Technician
Department of Radiation Oncology Fritz Eckstein, Ph.D.
University of North Carolina at Chapel Professor
Hill Max-Planck-Institute Fur
Chapel Hill, North Carolina Experimental Medizin
Gottingen, Germany
Lee E. Babiss
David A. Elkins, B.S.
Department of Cell Biology
Graduate Student
GLAXO
Center for Molecular Biology and Gene
Research Triangle Park, North Carolina
Therapy
Loma Linda University School of
C. Frank Bennett, Ph.D.
Medicine
Director Loma Linda, California
Department of Molecular Pharmacology
ISIS Pharmaceuticals Agamemnon A. Epenetos, Ph.D.
Carlsbad, California Professor
Department of Clinical Oncology
Jeffery S. Bishop Hammersmith Hospital
Department of Biology London, UK
Triplex Pharmaceutical Corporation
The Woodlands, Texas Mark A. Findeis, Ph.D.
Staff Scientist
Ellen P. Carmichael, Ph.D.
Department of Chemistry
Staff Scientist
Pharmaceutical Peptides, Inc.
The Immune Response Corporation
Cambridge, Massachusetts
Carlsbad, California
Nilabh Chaudhary, Ph.D. Daniel A. Geselowitz, Ph.D.
Senior Research Scientist Chemist
Department of Biology Department of Nuclear Medicine
Triplex Pharmaceutical Corporation NIH
The Woodlands, Texas Bethesda, Maryland
Alan M. Gewirtz, M.D. Eric T. Kool, Ph.D.
Associate Professor Assistant Professor
Department of Pathology and Internal Department of Chemistry
Medicine University of Rochester
University of Pennsylvania School of Rochester, New York
Medicine
David Kregenow, B.S.
Philadelphia, Pennsylvania
Medical Student
Calvin Stephen Roy Gooden, B.Sc. University of Pennsylvania School of
Imperial Cancer Research Fund Medicine
Oncology Unit Philadelphia, Pennsylvania
Hammersmith Hospital
Arthur M. Krieg, M.D.
London, UK
Assistant Professor
Charles J. Guinosso Department of Internal Medicine
ISIS Pharmaceuticals, Inc. University of Iowa
Carlsbad, California Iowa City, Iowa
Judith K. Guy-Caffey, Ph.D. and
Research Scientist Staff Physician
Department of Biology Department of Internal Medicine
Triplex Pharmaceutical Corporation Veterans Administration Medical Center
The Woodlands, Texas Iowa City, Iowa
Jeffery C. Hanvey Claude Malvy, Ph.D.
Department of Cell Biology Director of Research
GLAXO UR A 147
Research Triangle Park, North Carolina CNRS
Villejuif, France
Olaf Heidenreich, Ph.D.
Professor June Ray Merwin, Ph.D.
Max-Planck-Institute Fur Department of Cell Biology
Experimental Medizin The West Company
Gottingen, Germany Lionville, Pennsylvania
Jeffery Hughes, Ph.D. Marina Nechaeva
Assistant Professor Researcher
Department of Pharmaceutics Institute of Bioorganic Chemistry
University of Florida Novosibirsk, Russia
Gainesville, Florida
Leonard M. Neckers, Ph.D.
Krishna Jayaraman, Ph.D.
Head, Tumor Cell Section
Director of Oligonucleotide Chemistry
Clinical Pharmacology Branch
Department of Biology
NIH
Triplex Pharmaceutical Corporation
Bethesda, Maryland
The Woodlands, Texas
R.L. Juliano, Ph.D. Ludmila Pautova, Ph.D.
Professor and Chair Senior Researcher
Department of Pharmacology Institute of Bioorganic Chemistry
University of North Carolina at Chapel Novosibirsk, Russia
Hill
Mariusz Z. Ratajezak, Ph.D.
Chapel Hill, North Carolina
Senior Research Scientist
Valery Karamyshev, Ph.D. Department of Pathology
Researcher University of Pennsylvania School of
Institute of Bioorganic Chemistry Medicine
Novosibirsk, Russia Philadelphia, Pennsylvania
Catherine Ropert, Ph.D. Jamal Temsamani, Ph.D.
Research Scientist Associate Director
Department of Pharmaceutical Science Department of Discovery
Rhone Roulene Rorer Hybridon, Inc.
Vitry Cedex, France Worcester, Massachusetts
John J. Rossi, Ph.D.
Alain R. Thierry, Ph.D.
Associate Director
Visiting Scientist
Center for Molecular Biology and Gene
Lab Tumor Cell Biology
Therapy
NCI/NIH
Loma Linda University School of
Bethesda, Maryland
Medicine
and
Loma Linda, California
Adjunct Assistant Professor
Elena Rykova Department of Radiation Medicine
Researcher Georgetown University
Institute of Bioorganic Chemistry Washington, D.C.
Novosibirsk, Russia
Valentin V. Vlassov, Ph.D.
Henri M. Sasmor Professor
ISIS Pharmaceuticals, Inc. Institute of Bioorganic Chemistry
Carlsbad, California Novosibirsk, Russia
George L. Spitalny, Ph.D. Ernst Wagner, Ph.D.
Chief Scientific Officer Group Leader
The Immune Response Corporation Institute of Molecular Pathology
Carlsbad, California Vienna, Austria
and
Garry B. Takle, Ph.D.
Senior Scientist
Senior Scientist
Department of Therapeutic Vaccines
Innovir Labs, Inc.
Bender and Co.
New York, New York
Vienna, Austria
and
Adjunct Faculty Eric Wickstrom, Ph.D.
Department of Molecular Parasitology Professor
Rockefeller University Department of Pharmacology
New York, New York Thomas Jefferson University
Philadelphia, Pennsylvania
Jinyan Tang, Ph.D.
Director L.A.Yakubov, Ph.D.
Department of Process Chemistry Senior Researcher
Hybridon, Inc. Institute of Bioorganic Chemistry
Worcester, Massachusetts Novosibirsk, Russia
