ABSTRACT
Macromolecular prodrugs are developed as multiarmed agents against diverse viral pathogens. Lead candidates inhibit infectivity and replication of HIV, Ebola, influenza, measles, RSV, etc-thus being broad-spectrum antiviral agents.
Subject(s)
Antiviral Agents/pharmacology , Macromolecular Substances/chemistry , Polymers/chemistry , Prodrugs/pharmacology , Ribavirin/pharmacology , A549 Cells , Animals , Antiviral Agents/chemistry , Chick Embryo , HeLa Cells , Humans , Influenza A Virus, H1N1 Subtype/drug effects , Polyelectrolytes , Prodrugs/chemistry , Ribavirin/chemistry , Rous sarcoma virus/drug effectsABSTRACT
We report the first successful implementation of transfection agents to facilitate the delivery of non-nucleic acid based anti-inflammatory and anti-viral drugs. In doing so, we illustrate a new paradigm in the intracellular delivery of polyanionic drugs and also extend the scope and utility of successful tools of gene transfer into a new area of biomedical research.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/administration & dosage , Antiviral Agents/administration & dosage , Drug Delivery Systems/methods , Gene Transfer Techniques , Ribavirin/administration & dosage , Acrylic Resins/chemistry , Acrylic Resins/pharmacology , Anti-Inflammatory Agents, Non-Steroidal/chemistry , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Antiviral Agents/chemistry , Antiviral Agents/pharmacology , Cell Survival/drug effects , Humans , Lipopolysaccharides/antagonists & inhibitors , Lipopolysaccharides/pharmacology , Macrophages/drug effects , Macrophages/metabolism , Nitric Oxide/antagonists & inhibitors , Nitric Oxide/biosynthesis , Ribavirin/chemistry , Ribavirin/pharmacology , TransfectionABSTRACT
Efficacious, potent, and at the same time nontoxic macromolecular prodrugs of ribavirin are designed taking advantage over prodrug activation by the intracellular milieu. Activity of these prodrugs is illustrated in the cells hosting hepatitis C virus replication and also in the cells implicated in the inflammatory response to the viral infection.
Subject(s)
Antiviral Agents , Gene Expression Regulation, Viral/drug effects , Genome, Viral , Hepacivirus/metabolism , Hepatitis C/drug therapy , Polymethacrylic Acids , Prodrugs , Ribavirin , Antiviral Agents/chemical synthesis , Antiviral Agents/chemistry , Antiviral Agents/pharmacology , Cell Line , Hepacivirus/genetics , Hepatitis C/genetics , Hepatitis C/metabolism , Hepatitis C/pathology , Humans , Polymethacrylic Acids/chemical synthesis , Polymethacrylic Acids/chemistry , Polymethacrylic Acids/pharmacology , Prodrugs/chemical synthesis , Prodrugs/chemistry , Prodrugs/pharmacology , Ribavirin/analogs & derivatives , Ribavirin/chemical synthesis , Ribavirin/chemistry , Ribavirin/pharmacologyABSTRACT
Human immunodeficiency virus (HIV) and hepatitis C virus (HCV) represent tremendous healthcare burdens with a large proportion of patients hosting the two viruses at the same time. An altered hepatic function and immunity as well as cross-interference of drugs make treatment of co-infection increasingly challenging. Herein we report the first design of macromolecular prodrugs (MP) with concurrent success in fighting HIV and alleviating hepatitis (liver inflammation). To achieve this, polymer compositions were systematically screened in a broad range of molar mass and content of ribavirin - a broad spectrum antiviral agent. For the first time, we report that ribavirin is efficacious in fighting HIV and in the form of MP, the treatment is safe, both in terms of lack of association of ribavirin with red blood cells and lack of toxicity upon cellular internalization. The lead polymer compositions were also potent in anti-inflammatory assays with relevance to viral hepatitis - thus making up formulations with potential for treatment of co-infection with HIV and HCV.
ABSTRACT
Macromolecular prodrugs (MPs) are a powerful tool to alleviate side-effects and improve the efficacy of the broad-spectrum antiviral agent ribavirin. In this work, we sought an understanding of what makes an optimal formulation within the macromolecular parameter space--nature of the polymer carrier, average molar mass, drug loading, or a good combination thereof. A panel of MPs based on biocompatible synthetic vinylic and (meth)acrylic polymers was tested in an anti-inflammatory assay with relevance to alleviating inflammation in the liver during hepatitis C infection. Pristine polymer carriers proved to have a pronounced anti-inflammatory activity, a notion which may prove significant in developing MPs for antiviral and anticancer treatments. With conjugated ribavirin, MPs revealed enhanced activity but also higher toxicity. Therapeutic windows and therapeutic indices were determined and discussed to reveal the most potent formulation and those with optimized safety. Polymers were also tested as inhibitors of replication of the hepatitis C viral RNA using a subgenomic viral replicon system. For the first time, negatively charged polymers are revealed to have an intracellular activity against hepatitis C virus replication. Concerted activity of the polymer and ribavirin afforded MPs which significantly increased the therapeutic index of ribavirin-based treatment. Taken together, the systematic investigation of the macromolecular space identified lead candidates with high efficacy and concurrent direct activity against the hepatitis C virus and inflammation.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Antiviral Agents/pharmacology , Hepacivirus/drug effects , Hepatitis C/drug therapy , Macromolecular Substances/pharmacology , Prodrugs/pharmacology , Animals , Chemistry, Pharmaceutical , Drug Carriers , Hepatitis C/pathology , Hepatitis C/virology , Hepatocytes/drug effects , Hepatocytes/virology , In Vitro Techniques , Macrophages/drug effects , Mice , RNA, Viral/biosynthesis , Ribavirin/administration & dosage , Ribavirin/pharmacokinetics , Ribavirin/toxicity , Structure-Activity Relationship , Virus Replication/drug effectsABSTRACT
The release of azidothymidine from macromolecular prodrugs was designed to respond to the intracellular disulfide reshuffling. This drug has no thiol groups, and a response to this trigger was engineered using a self-immolative linker. The resulting formulations were fast-acting, efficacious, and highly potent with regards to suppressing the infectivity of the virus.
Subject(s)
Anti-HIV Agents/chemistry , Anti-HIV Agents/pharmacology , Disulfides/chemistry , HIV Infections/drug therapy , HIV-1/drug effects , Prodrugs/chemistry , Prodrugs/pharmacology , Zidovudine/pharmacology , Cell Line , Dose-Response Relationship, Drug , Humans , Macromolecular Substances/chemistry , Macromolecular Substances/pharmacology , Microbial Sensitivity Tests , Molecular Structure , Structure-Activity Relationship , Time Factors , Zidovudine/chemistryABSTRACT
Polymers in tune. Automated parallel polymer synthesis is developed to obtain libraries of macromolecular prodrugs of ribavirin, a broad-spectrum antiviral agent. As many as 10 identified lead polymer conjugates exhibit therapeutic efficacy matching that of the pristine drug and at the same time suppressed the origin of the main side effect of ribavirin.
Subject(s)
Prodrugs/chemistry , Prodrugs/pharmacology , Ribavirin/chemistry , Ribavirin/pharmacology , Animals , Antiviral Agents/chemistry , Antiviral Agents/pharmacology , Cell Line , Chromatography, Gel , Drug Discovery , Macrophages/drug effects , Macrophages/metabolism , Magnetic Resonance Spectroscopy , Mice , Molecular Weight , Nitric Oxide/analysis , Nitric Oxide/metabolismABSTRACT
Ribavirin (RBV)-containing polymers are synthesized based on poly(N-vinylpyrrolidone) and poly(acrylic acid), two polymers with extensive characterization in biomedicine. The copolymers are shown to exhibit a minor to negligible degree of association with erythrocytes, thus effectively eliminating the origin of the main side effects of RBV. The therapeutic benefit of macromolecular RBV prodrugs is illustrated by matched efficacy in suppressing production of nitric oxide by stimulated cultured macrophages as compared to pristine RBV with no associated cytotoxicity, which is in stark contrast to an RBV-based treatment which results in a significant decrease in cell viability. These results contribute to the development of antiviral polymer therapeutics and delivery of RBV in particular.
Subject(s)
Antiviral Agents/administration & dosage , Prodrugs/chemistry , Prodrugs/pharmacology , Ribavirin/administration & dosage , Acrylic Resins/chemistry , Cell Survival/drug effects , Chemistry Techniques, Synthetic , Dose-Response Relationship, Drug , Erythrocytes/drug effects , Hep G2 Cells/drug effects , Humans , Macrophages/drug effects , Nitric Oxide/metabolism , Polyvinyls/chemistry , Prodrugs/chemical synthesis , Pyrrolidinones/chemistryABSTRACT
Ribavirin (RBV), a broad-spectrum antiviral agent, is a standard medication against hepatitis C virus (HCV). However, despite the decades of clinical success, the mechanism of action of this drug against HCV remains a subject of debate. Furthermore, the appeal of this therapeutic agent is considerably lessened by unfavorable pharmacokinetics. This interdisciplinary study contributes to the understanding of intracellular effects exerted by RBV and presents a successful design of macromolecular prodrugs of RBV to achieve a safer treatment. Specifically, we demonstrate that RBV exhibits a pronounced anti-inflammatory activity in cultured macrophages as is evidenced by a 2-fold decrease in the levels of produced nitric oxide achieved using a clinically relevant concentration of this drug. However, this effect was characterized by a rather narrow therapeutic window with experimental values of EC50 and IC50 being 7 and 19 µM, respectively. Macromolecular prodrugs were obtained using an acrylate derivative of RBV, RAFT polymerization technique, and N-vinyl pyrrolidone as a partner monomer. The synthesized polymers were characterized with uniform molecular weights, relatively narrow polydispersities, and gradually increasing content of RBV. The resulting polymer therapeutics were effective in delivering their payload to the cultured macrophages and afforded a significantly wider therapeutic window, as much as >1000 µM (18-fold in relative values). Taken together, this work contributes significantly to the development of safer methods for delivery of RBV, as well as understanding the mechanism of action and origins of the side effects of this broad-spectrum antiviral agent.
Subject(s)
Antiviral Agents/pharmacology , Nitric Oxide/biosynthesis , Prodrugs/pharmacology , Ribavirin/pharmacology , Animals , Antiviral Agents/chemical synthesis , Antiviral Agents/chemistry , Cell Survival/drug effects , Cells, Cultured , Dose-Response Relationship, Drug , Drug Design , Hep G2 Cells , Humans , Macromolecular Substances/chemical synthesis , Macromolecular Substances/chemistry , Macromolecular Substances/pharmacology , Macrophages/drug effects , Macrophages/metabolism , Mice , Molecular Structure , Prodrugs/chemical synthesis , Prodrugs/chemistry , Ribavirin/chemistry , Structure-Activity Relationship , Time FactorsABSTRACT
Hydrogel nanoparticles (HNP) are an emerging tool of biomedicine with unique materials characteristics, scope, and utility. These hydrated, soft colloidal carriers can penetrate through voids with dimensions narrower than the size of the particle, provide stabilization for fragile biological cargo and allow diffusion and exchange of solutes with external phase. However, techniques to assemble HNP are few; solitary examples exist of biocompatible polymers being formulated into HNP; and knowledge on the biomedical properties of HNP remains rather cursory. In this work, we investigate assembly of HNP based on a polymer with decades of prominence in the biomedical field, poly(vinyl alcohol), PVA. We develop a novel method for production of PVA HNP through nanoprecipitation-based assembly of polymer nanoparticles and subsequent physical hydrogelation of the polymer. Polymer nanoparticles and HNP were visualized using scanning electron microscopy and fluorescence imaging, and characterized using dynamic light scattering and zeta potential measurements. Interaction of PVA HNP with mammalian cells was investigated using flow cytometry, viability screening, and measurements of nitric oxide production by cultured macrophages. The latter analyses revealed that PVA administered as a polymer solution or in the form of HNP resulted in no measurable increase in production of the inflammation marker. Unexpectedly, PVA HNP exerted a pronounced inhibition of NO synthesis by stimulated macrophages, that is, had an anti-inflammatory activity. This effect was accomplished with a negligible change in the cell viability and was not observed when PVA was administered as a polymer solution. To the best of our knowledge, this is the first observation of inhibition of NO synthesis in macrophages by administered nanoparticles and specifically hydrogel nanoparticles. Taken together, our results present PVA HNP as promising colloidal hydrogel nanocarriers for biomedical applications, specifically drug delivery and assembly of intracellular biosensors.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/chemistry , Drug Carriers/chemistry , Macrophages/drug effects , Nanoparticles/chemistry , Nitric Oxide/antagonists & inhibitors , Polyvinyl Alcohol/chemistry , Animals , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Cell Line , Cell Survival/drug effects , Drug Carriers/pharmacology , Hep G2 Cells , Humans , Hydrogels , Lipopolysaccharides/pharmacology , Macrophages/cytology , Macrophages/metabolism , Mice , Nanoparticles/ultrastructure , Nitric Oxide/biosynthesis , Particle Size , Polyvinyl Alcohol/pharmacologyABSTRACT
Chemi-enzymatic synthesis of ribavirin acrylate and subsequent RAFT co-polymerization with acrylic acid afforded a formulation of a broad spectrum antiviral drug which avoids accumulation in erythrocytes, the origin of the main side effect of ribavirin. In cultured macrophages the macromolecular prodrugs exhibited decreased toxicity while maintaining the anti-inflammatory action of ribavirin.
Subject(s)
Acrylates/pharmacology , Macrophages/drug effects , Prodrugs/pharmacology , Ribavirin/pharmacology , Acrylates/chemical synthesis , Acrylates/chemistry , Animals , Cells, Cultured , Dose-Response Relationship, Drug , Macromolecular Substances/chemical synthesis , Macromolecular Substances/chemistry , Macromolecular Substances/pharmacology , Macrophages/metabolism , Mice , Molecular Conformation , Nitric Oxide/antagonists & inhibitors , Nitric Oxide/biosynthesis , Prodrugs/chemical synthesis , Prodrugs/chemistry , Ribavirin/chemical synthesis , Ribavirin/chemistry , Structure-Activity RelationshipABSTRACT
Due to its versatility and ease of use the layer-by-layer (LbL) assembly technique has been under intensive investigation for drug and gene delivery applications. Especially the development of responsive LbL materials has advanced significantly in recent years. Responsiveness plays an important role in many delivery applications, either for loading of therapeutics or controlled and triggered release. In general four basic mechanisms within responsive LbL films have been identified: disruption of layer interactions, degradation of the LbL film, multilayer destruction via physical stimuli, and phase transitions or polymer rearrangements within the LbL film. This review will outline these different mechanisms and highlight recent advances in these fields.