Download or read book Peptide Conjugation to Enhance Oligonucleotide Delivery written by Justin Mahoney Wolfe and published by . This book was released on 2018 with total page 419 pages. Available in PDF, EPUB and Kindle. Book excerpt: The intracellular delivery of functional macromolecules remains an outstanding challenge in biomedicine. While small molecules can diffuse through the plasma membrane, many large therapeutic molecules are not internalized to an appreciable extent. One strategy to improve cell uptake involves linking the molecule of interest to a cell-penetrating peptide (CPP). CPPs are widely employed to enhance macromolecule delivery, with hundreds of different peptides and modifications reported to improve cellular uptake. In this thesis, CPPs were systematically investigated and chemically altered to facilitate the delivery of antisense oligonucleotides. To accurately compare the existing CPPs, 64 CPP sequences were synthesized, conjugated to oligonucleotides, and assayed for delivery. These CPPs showed a range of effectiveness, with some CPPs hindering the delivery of oligonucleotide cargo and others leading to a 10-fold increase in oligonucleotide activity. To help identify which CPPs might be valuable for oligonucleotide delivery specifically, a computational model was developed to predict, de novo, whether or not a CPP will be effective. When experimentally validated, this model successfully predicted which sequences would improve oligonucleotide delivery greater than 3-fold. Multiple strategies were employed to improve CPP effectiveness. First, arginine-rich CPPs were chemically modified with perfluoroarenes. Cyclic and bicyclic CPPs were synthesized by linking multiple cysteine residues together with a perfluoroarene. After oligonucleotide conjugation, these peptides led to a 14-fold increase in delivery. Second, two different CPPs were combined into one long chimeric sequence. The CPP chimeras were highly active, leading to a 20-fold increase in oligonucleotide delivery. Third, the idea of combining multiple CPPs led to the development of a method for the rapid synthesis combinatorial peptide conjugates. Using the judicious choice of bioconjugation chemistry, highly-active modular constructs were synthesized that contain three peptides linked to one oligonucleotide. In addition to CPPs for oligonucleotide delivery, one section of this thesis employed perfluoroaryl macrocyclic peptides to address the challenge of peptide delivery across the blood-brain barrier. An additional section developed a new peptide conjugation strategy that uses palladium-peptide oxidative addition complexes as solid, storable, and water-soluble reagents for bioconjugation.
Download or read book Therapeutic Oligonucleotides written by Jens Kurreck and published by Royal Society of Chemistry. This book was released on 2008 with total page 362 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book provides a compelling overall update on current status of RNA interference
Download or read book Novel Approaches to Oligonucleotide Conjugation to a Cell Penetrating Tat Peptide written by Victoria Steven and published by . This book was released on 2009 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Modifed oligonucleotides are routinely employed as bioanalytical probes for use in diagnostics; however, a major limiting factor in oligonucleotide-based diagnostics is poor cellular uptake. This can be overcome by covalent attachment of a cell penetrating peptide. Diels-Alder cycloaddition is an attractive methodology for oligonucleotide peptide conjugation; the reaction is fast and chemoselective and the reaction rate is greatly enhanced in aqueous media. An oligonucleotide sequence has been derivatised with a series of dienes at the 5'- terminus, through chemical synthesis of a series of unique dienyl modified phosphoramidites. Investigation into the effect of diene type on the efficiency of conjugation to a maleimido derivatised Tat peptide derivative, via the Diels-Alder cycloaddition, has been performed. The optimal diene for biomolecule conjugation was found to be cyclohexadiene in the conjugation of oligonucleotides to a cell penetrating peptide via Diels-Alder cycloaddition for the first time. An oligonucleotide sequence has also been derivatised with cyclohexadiene internally; Diels-Alder cycloadditions of these oligonucleotides to a maleimido derivatised Tat peptide derivative were also successful. However, oligonucleotide conjugation to Tat peptide via Diels-Alder cycloaddition of fluorescently labelled oligonucleotides for visualisation in cell studies has not been as successful as for unlabelled oligonucleotides. Generation of a biocatalytic DNA sequence for the acceleration of Diels-Alder cycloadditions has been attempted. Through a SELEX-type process, a DNA sequence has been isolated for experimental determination of its potential as a biological catalyst for Diels-Alder cycloadditions. Oligonucleotide conjugation to Tat peptide via gold nanoparticles has been achieved. Chemical synthesis of a dithiolated, long-chain PEGylated ligand allowed covalent attachment of Tat peptide to gold nanoparticles. Bifunctionalisation of gold nanoparticles with oligonucleotides and this ligand, followed by covalent attachment of Tat peptide generated the desired conjugate. The conjugates have been shown to hybridise successfully with the complementary oligonucleotide sequence, thereby displaying potential for their use as bioanalytical probes. Methods for quantification of both oligonucleotides and Tat peptide conjugated to gold nanoparticles, by enzyme hydrolysis, have been developed.
Download or read book Nanoscale Graphitic Carbon Nitride written by Alagarsamy Pandikumar and published by Elsevier. This book was released on 2021-09-09 with total page 570 pages. Available in PDF, EPUB and Kindle. Book excerpt: Nanoscale Graphitic Carbon Nitride focuses on multi-functional applications including energy conversion, storage and healthcare. Polymeric graphitic carbon nitride materials have attracted much attention in recent years because of their similarity to graphene. They are composed of carbon, nitrogen and some minor hydrogen content. In contrast to graphene, g-Graphitic carbon nitride is a medium band-gap semiconductor and in that role an effective photocatalyst and chemical catalyst for a broad variety of reactions and applications. This book covers the fundamentals and applications of graphitic carbon nitride (g-C3N4) in different sectors. It also covers the application of graphitic carbon nitride-based composites with metal, metal oxides, metal sulphide and carbon-based materials. This is an important resource for researchers in the fields of materials science, engineering, energy storage and chemical engineering who want to understand how nanoscale graphitic carbon nitride is being used for a range of industrial applications and processes. Outlines the major properties of nanoscale graphitic carbon nitride, along with their major application areas Assesses the challenges of manufacturing graphitic carbon nitride on a mass scale Explains major synthesis methods for nanoscale graphitic carbon nitride
Download or read book Simple and Double Conjugates of Peptides and Oligonucleotides written by Jordi Agramunt Pi and published by . This book was released on 2019 with total page 344 pages. Available in PDF, EPUB and Kindle. Book excerpt: "2,2-Disubstituted cyclopent-4-ene-1,3-diones (CPDs), which have been as non-hydrolysable maleimide analogs, have been found to possess an unexpected reactivity with N-terminal cysteines. Whilst maleimides react in an irreversible manner with all types of thiol nucleophiles, the Michael-type addition between CPDs and N-terminal cysteines, furnish a stable product with a mass 20 Da lower than the expected Michael-type adduct. This newly found reactivity can be applied for different purposes. In first instance, CPD-derivatized peptides can be used to synthesize conjugates by reaction with cystine-derivatized biomolecules such as peptides, peptide nucleic acids (PNAs) or oligonucleotides. In addition, the combination of the CPD-Cys reaction with other "click" reactions has been explored. In the first instance, the CPD-Cys reaction was paired the thia-Michael addition of thiols to maleimides using peptides containing two cysteines (one at the N-terminus). By first performing a CPD-Cys reaction at the N-terminus and then the addition of a maleimide, double conjugation could be easily achieved. In another case, combinations with oxime ligation and copper(I) catalyzed azide-alkyne (CuAAC) cycloadditions have been accomplished with a different degree of success. In the latter case, the CPD-Cys reaction has to be strictly performed before the CuAAC. In a further examination why this was the case, it was found out that CPDs react with azides to furnish two stable compounds one is the product of a 1,3-dipolar cycloaddition and the other results from nitrogen loss. Secondly, the bioconjugation of a splice-switching enhancing molecule known as "Retro-1" to an oligonucleotide with splice-switching activity described as 623 has been explored. In this part of the work, the complete synthesis of described Retro-1 has been accomplished in addition to that of other several analogues containing either a 1,3-diene, a thiol or a phosphoramidite group appending from two positions, nitrogen 4 or carbon 3. For this purpose, two synthetic strategies were followed one making use of valuable intermediates obtained from the Retro-1 synthesis and the other utilizing a properly protected lysine analogue to construct the Retro scaffold and introduce a reactive group at carbon 3. Additionally to all these reactants two oligonucleotides were synthesized: one with the 623 sequence and another with the same nucleobases but different order coined "scrambled". Both oligonucleotides were appended at the 5' position with a protected maleimide phosphoramidite (to obtain the corresponding maleimido-oligonucleotide and allow a Diels-Alder or thia-Michael addition to be performed) or the Retro-phosphoramidite. Finally, in this work the possibility of using 7-oxanorbornenes as dienophiles in the inverse electron-demanding Diels-Alder cycloaddition (IEDDA) with 3,6-disbustituted 1,2,4,5-tetrazines has been explored. The oxanorbornene moiety has been appended at the N-terminus and internal positions in peptides, and at the 5' and 3' positions of oligonucleotides utilizing solid-phase protocols. For the tetrazine counterpart optimization processes have been followed for the synthesis of N-terminal tetrazine peptides and for derivatization with biologically relevant molecules in reasonably good success. Once oxanorbornene- and tetrazine-containing derivatives were obtained it was observed that the IEDDA reaction took place satisfactorily, allowing for the synthesis of peptide and oligonucleotide conjugates with small molecules. In second term, the possibility of double conjugation involving at least one IEDDA reaction has been explored. The IEDDA has been combined with the Diels-Alder cycloaddition between a maleimide and a 1,3-diene, the thia-Michael between a thiol and a maleimide, the CPD-Cys reaction (utilizing CPDs and N-terminal cysteines as previously commented) and the aromatic nucleophilic substitution between a thiol and a chlorotetrazines. In every combination tested the double conjugate target compound was obtained, usually performing both reactions simultaneously, but sometimes in a "one-pot" fashion, with the sequential addition of reactants due to side reactions. In one case, it was necessary to isolate the intermediate monoconjugate because of difficulties related to undesired interferences between reactants." -- TDX.
Download or read book Bioconjugate Techniques written by Greg T. Hermanson and published by Academic Press. This book was released on 2010-07-26 with total page 1233 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bioconjugate Techniques, 2nd Edition, is the essential guide to the modification and cross linking of biomolecules for use in research, diagnostics, and therapeutics. It provides highly detailed information on the chemistry, reagent systems, and practical applications for creating labeled or conjugate molecules. It also describes dozens of reactions with details on hundreds of commercially available reagents and the use of these reagents for modifying or cross linking peptides and proteins, sugars and polysaccharides, nucleic acids and oligonucleotides, lipids, and synthetic polymers. A one-stop source for proven methods and protocols for synthesizing bioconjugates in the lab Step-by-step presentation makes the book an ideal source for researchers who are less familiar with the synthesis of bioconjugates More than 600 figures that visually describe the complex reactions associated with the synthesis of bioconjugates Includes entirely new chapters on the latest areas in the field of bioconjugation as follows: Microparticles and nanoparticlesSilane coupling agentsDendrimers and dendronsChemoselective ligationQuantum dotsLanthanide chelatesCyanine dyesDiscrete PEG compoundsBuckyballs,fullerenes, and carbon nanotubesMass tags and isotope tagsBioconjugation in the study of protein interactions
Download or read book Oligonucleotide Based Drugs and Therapeutics written by Nicolay Ferrari and published by John Wiley & Sons. This book was released on 2018-07-31 with total page 576 pages. Available in PDF, EPUB and Kindle. Book excerpt: A comprehensive review of contemporary antisense oligonucleotides drugs and therapeutic principles, methods, applications, and research Oligonucleotide-based drugs, in particular antisense oligonucleotides, are part of a growing number of pharmaceutical and biotech programs progressing to treat a wide range of indications including cancer, cardiovascular, neurodegenerative, neuromuscular, and respiratory diseases, as well as other severe and rare diseases. Reviewing fundamentals and offering guidelines for drug discovery and development, this book is a practical guide covering all key aspects of this increasingly popular area of pharmacology and biotech and pharma research, from the basic science behind antisense oligonucleotides chemistry, toxicology, manufacturing, to safety assessments, the design of therapeutic protocols, to clinical experience. Antisense oligonucleotides are single strands of DNA or RNA that are complementary to a chosen sequence. While the idea of antisense oligonucleotides to target single genes dates back to the 1970's, most advances have taken place in recent years. The increasing number of antisense oligonucleotide programs in clinical development is a testament to the progress and understanding of pharmacologic, pharmacokinetic, and toxicologic properties as well as improvement in the delivery of oligonucleotides. This valuable book reviews the fundamentals of oligonucleotides, with a focus on antisense oligonucleotide drugs, and reports on the latest research underway worldwide. • Helps readers understand antisense molecules and their targets, biochemistry, and toxicity mechanisms, roles in disease, and applications for safety and therapeutics • Examines the principles, practices, and tools for scientists in both pre-clinical and clinical settings and how to apply them to antisense oligonucleotides • Provides guidelines for scientists in drug design and discovery to help improve efficiency, assessment, and the success of drug candidates • Includes interdisciplinary perspectives, from academia, industry, regulatory and from the fields of pharmacology, toxicology, biology, and medicinal chemistry Oligonucleotide-Based Drugs and Therapeutics belongs on the reference shelves of chemists, pharmaceutical scientists, chemical biologists, toxicologists and other scientists working in the pharmaceutical and biotechnology industries. It will also be a valuable resource for regulatory specialists and safety assessment professionals and an important reference for academic researchers and post-graduates interested in therapeutics, antisense therapy, and oligonucleotides.
Download or read book Innovations for Next Generation Antibody Drug Conjugates written by Marc Damelin and published by Springer. This book was released on 2018-05-29 with total page 358 pages. Available in PDF, EPUB and Kindle. Book excerpt: Antibody-drug conjugates (ADCs) stand at the verge of a transformation. Scores of clinical programs have yielded only a few regulatory approvals, but a wave of technological innovation now empowers us to overcome past technical challenges. This volume focuses on the next generation of ADCs and the innovations that will enable them. The book inspires the future by integrating the field’s history with novel strategies and cutting-edge technologies. While the book primarily addresses ADCs for solid tumors, the last chapter explores the emerging interest in using ADCs to treat other diseases. The therapeutic rationale of ADCs is strong: to direct small molecules to the desired site of action (and away from normal tissues) by conjugation to antibodies or other targeting moieties. However, the combination of small and large molecules imposes deep complexity to lead optimization, pharmacokinetics, toxicology, analytics and manufacturing. The field has made significant advances in all of these areas by improving target selection, ADC design, manufacturing methods and clinical strategies. These innovations will inspire and educate scientists who are designing next-generation ADCs with the potential to transform the lives of patients.
Download or read book Design and Delivery of SiRNA Therapeutics written by Henrik J. Ditzel and published by . This book was released on 2021 with total page 469 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume details protocols on rationale design of therapeutic siRNA molecules and its encapsulation with smart vehicles to overcome the barriers to an effective administration in vivo. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Design and Delivery of SiRNA Therapeutics aims to ensure successful results in the further study of this vital field. This volume details protocols on rationale design of therapeutic siRNA molecules and its encapsulation with smart vehicles to overcome the barriers to an effective administration in vivo. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Design and Delivery of SiRNA Therapeutics aims to ensure successful results in the further study of this vital field.
Download or read book Nucleic Acid Drugs written by Akira Murakami and published by Springer. This book was released on 2012-07-04 with total page 189 pages. Available in PDF, EPUB and Kindle. Book excerpt: New Antisense Strategies: Chemical Synthesis of RNA Oligomers, by Junichi Yano und Gerald E. Smyth Development and Modification of Decoy Oligodeoxynucleotides for Clinical Application, by Mariana Kiomy Osako, Hironori Nakagami und Ryuichi Morishita Modulation of Endosomal Toll-Like Receptor-Mediated Immune Responses by Synthetic Oligonucleotides, by Ekambar R. Kandimalla und Sudhir Agrawal Delivery of Nucleic Acid Drugs, by Yan Lee und Kazunori Kataoka Aptamer: Biology to Applications, by Yoshikazu Nakamura Development and Clinical Applications of Nucleic Acid Therapeutics, by Veenu Aishwarya, Anna Kalota und Alan M. Gewirtz
Download or read book Peptide Nucleic Acids written by Peter E. Nielsen and published by Humana Press. This book was released on 2002-07-23 with total page 274 pages. Available in PDF, EPUB and Kindle. Book excerpt: Peptide nucleic acids (PNAs) have now existed for slightly more than ten years, with the interest in and applications of this pseudopeptide DNA mimic steadily increasing during the entire period. PNAs have rapidly attracted the attention of scientists from a diversity of fields ranging from (bio)organic and biophysical chemistry to prebiotic evolution, and from molecular biology to genetic diagnostics and drug development. Many of the applications take advantage of the unique properties of PNA—an uncharged pseudopeptide—that distinguish this DNA mimic from more traditional DNA analogs. Rather than trying to create a comprehensive collection of all published methods and protocols involving PNA—many of which have not yet been validated— I have decided to concentrate on select protocols that are either very well established by several groups around the world, such as PCR-clamping and in situ hybridization, or on new methods that may have broader future impact. Basic methods for PNA oligomer synthesis and analyses have also been included. I am very grateful to those friends and colleagues who have enthusiastically contributed their work, discussions, and writing, and thereby made this book possible. Peter E. Nielsen v Contents Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix IINTRODUCTION 1 PNA Technology Peter E. Nielsen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 II CHEMISTRY 2 Solid Phase Synthesis of PNA Oligomers Frederik Beck. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 3 Synthesis of PNA-Peptide Conjugates Satish Kumar Awasthi and Peter E. Nielsen. . . . . . . . . . . . . . . . . . 43 4 Parallel Synthesis of PNA-Peptide Conjugate Libraries Satish Kumar Awasthi and Peter E. Nielsen. . . . . . . . . . . . . . . . . .
Download or read book Advanced Healthcare Materials written by Ashutosh Tiwari and published by John Wiley & Sons. This book was released on 2014-05-09 with total page 421 pages. Available in PDF, EPUB and Kindle. Book excerpt: Offers a comprehensive and interdisciplinary view of cutting-edge research on advanced materials for healthcare technology and applications Advanced healthcare materials are attracting strong interest in fundamental as well as applied medical science and technology. This book summarizes the current state of knowledge in the field of advanced materials for functional therapeutics, point-of-care diagnostics, translational materials, and up-and-coming bioengineering devices. Advanced Healthcare Materials highlights the key features that enable the design of stimuli-responsive smart nanoparticles, novel biomaterials, and nano/micro devices for either diagnosis or therapy, or both, called theranostics. It also presents the latest advancements in healthcare materials and medical technology. The senior researchers from global knowledge centers have written topics including: State-of-the-art of biomaterials for human health Micro- and nanoparticles and their application in biosensors The role of immunoassays Stimuli-responsive smart nanoparticles Diagnosis and treatment of cancer Advanced materials for biomedical application and drug delivery Nanoparticles for diagnosis and/or treatment of Alzheimers disease Hierarchical modelling of elastic behavior of human dental tissue Biodegradable porous hydrogels Hydrogels in tissue engineering, drug delivery, and wound care Modified natural zeolites Supramolecular hydrogels based on cyclodextrin poly(pseudo)rotaxane Polyhydroxyalkanoate-based biomaterials Biomimetic molecularly imprinted polymers
Download or read book Drug Delivery written by Binghe Wang and published by John Wiley & Sons. This book was released on 2016-04-18 with total page 740 pages. Available in PDF, EPUB and Kindle. Book excerpt: Following its successful predecessor, this book covers the fundamentals, delivery routes and vehicles, and practical applications of drug delivery. In the 2nd edition, almost all chapters from the previous are retained and updated and several new chapters added to make a more complete resource and reference. • Helps readers understand progress in drug delivery research and applications • Updates and expands coverage to reflect advances in materials for delivery vehicles, drug delivery approaches, and therapeutics • Covers recent developments including transdermal and mucosal delivery, lymphatic system delivery, theranostics • Adds new chapters on nanoparticles, controlled drug release systems, theranostics, protein and peptide drugs, and biologics delivery
Download or read book Oligonucleotides and Analogues written by Fritz Eckstein and published by Oxford University Press, USA. This book was released on 1991 with total page 346 pages. Available in PDF, EPUB and Kindle. Book excerpt: Interest in oligonucleotide synthesis has expanded dramatically since the publication of M. Gait's book in this series in 1984. The process has been automated, new reagents have been developed, and the range of uses for the compounds produced has increased and continues to grow. This volume provides practical guidance on oligonucleotide synthesis and methods for introducing modifications into these molecules. State- of-the-art techniques for automated synthesis are covered in two chapters on the production of oligodeoxynucleotides and oligoribonucleotides. Synthesis of modified oligodeoxynucleotides is also detailed, including modification of the phosphate backbone to produce phosphorothioates, phosphorodithiolates and methyl phosphonates, all of which are of considerable importance due to thier potential therapeutic applications. Other chapters decsribe production of sugar-modified ilgodeoxynucleotides and the attachment of various reported groups; these techniques are useful to those interested in non-radioactive probes for hybridization and for the study of DNA-DNA and DNA protein interactions. This book will be of interest to academic and industrial researchers, enabling both chemists and non-chemists to synthesize oligonucleotides and analogues for a wide variety of experimental purposes.
Download or read book Modified Nucleic Acids written by Kazuhiko Nakatani and published by Springer. This book was released on 2016-04-04 with total page 280 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book spans diverse aspects of modified nucleic acids, from chemical synthesis and spectroscopy to in vivo applications, and highlights studies on chemical modifications of the backbone and nucleobases. Topics discussed include fluorescent pyrimidine and purine analogs, enzymatic approaches to the preparation of modified nucleic acids, emission and electron paramagnetic resonance (EPR) spectroscopy for studying nucleic acid structure and dynamics, non-covalent binding of low- and high-MW ligands to nucleic acids and the design of unnatural base pairs. This unique book addresses new developments and is designed for graduate level and professional research purposes.
Download or read book Galanin written by Tomas Hökfelt and published by Springer Science & Business Media. This book was released on 2010-08-18 with total page 285 pages. Available in PDF, EPUB and Kindle. Book excerpt: Galanin is a neuropeptide found both in the central and peripheral nervous system. The 29-amino acid peptide (named after its N-terminal glycine and C-terminal alanine) was identified in 1983 by its C-terminal amidation. This 'reverse' approach, that is to discover a substance through a distinct chemical feature, and only subsequently to characterize its biological activity, was novel and has been successful in the identification of several other peptides. After the structure of galanin was determined in 1983, functional studies were performed with material purified from natural sources until the synthetic form of the peptide became available. Galanin can act as transmitter, modulator and trophic factor, and is involved in a number of physiological processes such as hormone secretion, cardiovascular mechanisms, feeding and cognition. This peptide may also be of significance for a number of pathological processes/disorders including pain, depression, Alzheimer's disease, epilepsy, addiction and cancer. This wide diversity of actions is mediated by three galanin receptor subtypes. The studies reviewed in this volume give a fairly complete overview of the spectrum of the biological actions and functions of galanin and its receptors and on possible therapeutic applications in a number of pathological conditions.
Download or read book Novel Methodology for the Linking of Oligonucleotide peptide Conjugates written by Jason Robert Epstein and published by . This book was released on 1999 with total page 136 pages. Available in PDF, EPUB and Kindle. Book excerpt:
