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1.
J Virol ; 96(22): e0150222, 2022 11 23.
Artigo em Inglês | MEDLINE | ID: mdl-36314826

RESUMO

Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract infections in children. In humans, natural infection with RSV affords only partial long-term protection from reinfection, and there is no licensed RSV vaccine currently available. We have developed a new vaccine candidate, termed RSVNanoVax, composed of polyanhydride nanoparticles encapsulating the RSV prefusion F protein and a CpG 1668 oligodeoxynucleotide adjuvant. We recently reported that vaccination of inbred BALB/c mice with RSVNanoVax induced both RSV-specific cellular and humoral immunity, which provided protection from viral replication and RSV-induced disease. To further assess the efficacy of RSVNanoVax, here, we utilized outbred Swiss Webster mice to examine vaccine efficacy in a more genetically diverse population. Following intranasal prime-boost vaccination with RSVNanoVax, Swiss Webster mice exhibited robust titers of systemic RSV F-directed IgG antibodies and RSV F-directed IgA within the lungs and nasal passages that were sustained out to at least 1 year post-vaccination. Serum antibodies maintained robust neutralizing activity against both RSV A and B strains. Following RSV challenge, vaccinated Swiss Webster mice exhibited rapid viral clearance from the lungs. Overall, our results indicate that RSVNanoVax represents a promising RSV vaccine candidate capable of providing long-term protection and immunity in a genetically diverse population. IMPORTANCE Respiratory syncytial virus (RSV) infection causes thousands of infections and deaths in children and elderly adults each year. Research in this field is of great importance as there remains no licensed vaccine to prevent RSV infections. We developed a novel vaccine candidate, RSVNanoVax, utilizing the RSV prefusion F protein encapsulated in polyanhydride nanoparticles. Here, we show that the intranasal delivery of RSVNanoVax protected outbred mice from viral replication within the lungs when challenged with RSV out to 1 year post-vaccination. Additionally, RSV-specific antibody responses were generated in both the serum and lung tissue and sustained long-term. These results demonstrate that our vaccine is an encouraging candidate for driving long-term protection in the lungs in a genetically diverse population.


Assuntos
Infecções por Vírus Respiratório Sincicial , Vacinas contra Vírus Sincicial Respiratório , Animais , Humanos , Camundongos , Anticorpos Antivirais/sangue , Modelos Animais de Doenças , Imunoglobulina G/sangue , Camundongos Endogâmicos BALB C , Polianidridos , Infecções por Vírus Respiratório Sincicial/prevenção & controle , Vacinas contra Vírus Sincicial Respiratório/imunologia , Vírus Sincicial Respiratório Humano , Proteínas Virais de Fusão , Anticorpos Neutralizantes/sangue , Nanopartículas , Administração Intranasal
2.
Int J Mol Sci ; 23(19)2022 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-36232762

RESUMO

Polyanhydrides based on betulin are promising materials for use in controlled drug delivery systems. Due to the broad biological activity of betulin derivatives and lack of toxicity in vitro and in vivo, these polymers can be used both as polymeric prodrug and as carriers of other biologically active compounds. In this study, we develop a novel amphiphilic branched polyanhydrides synthesized by the two-step melt polycondensation of betulin disuccinate (DBB) and a tricarboxylic derivative of poly(ethylene glycol) (PEG_COOH). DBB and PEG_COOH were used as the hydrophobic and hydrophilic segments, respectively. The content of DBB in copolymers was from 10 to 95 wt%. Copolymers were assessed for their cytostatic activity against various cancer cell lines. Compared to linear DBB and PEG-based polyanhydrides, the branched polyanhydrides exhibited higher anticancer activity. The obtained polymers were able to self-assemble in water to form micelles with hydrodynamic diameters from 144.8 to 561.8 nm. and are stable over a concentration range from 12.5 µg/mL to 6.8 mg/mL. The formed micelles were found to be spherical in shape using a scanning electron microscope. It was found that the structure and composition of polyanhydrides affected the hydrodynamic diameter of the micelles. The branched betulin-based polyanhydrides have the potential to serve as biodegradable polymer prodrugs or carriers for other bioactive compounds.


Assuntos
Citostáticos , Nanopartículas , Polianidridos , Pró-Fármacos , Portadores de Fármacos/química , Micelas , Nanopartículas/química , Polietilenoglicóis/química , Triterpenos , Água
3.
Biomacromolecules ; 23(8): 3417-3428, 2022 08 08.
Artigo em Inglês | MEDLINE | ID: mdl-35881559

RESUMO

The alternating architecture and hydrophobic side chains hinder hydrolytic cleavage and anhydride interchange in poly(sebacic acid-ricinoleic acid) (P(SA-RA)), which provides stable polyanhydrides at room temperature. In this report, a series of polyanhydrides were designed to investigate the effect of ester bonds, hydrophobic side chains, phenyl moieties, and their distance from anhydride bonds on their stability and properties. Polyanhydrides with alternating architecture are constructed by the polymerization of ester-diacids prepared from ricinoleic or other hydroxy acids with anhydrides such as succinic, maleic, and phthalic anhydrides. The hydrophobic side chains are designed closer to anhydride bonds to investigate hindrance to hydrolytic cleavage and anhydride interchange. Polyanhydrides were obtained by the activation of ester-diacid using acetic anhydride followed by melt condensation. The reactions were monitored by NMR, Fourier transform infrared (FTIR), and gel permeation chromatography (GPC). The synthesized poly(ester-anhydride)s with a shorter chain length compared to P(SA-RA) were stable at room temperature. The hydrolytic degradation studies reveal that the phenyl moiety present in poly(ricinoleic acid phthalate) (PRAP) and poly(hydroxystearic acid phthalate) (PHSAP) reduces the hydrolysis of anhydride bonds. Poly(hydroxyoctanoic acid succinate) (PHOAS) demonstrates the highest molecular weight of all tested polymers. The results reveal that the presence of hydrophobic side chains, phenyl moieties, and their distance from anhydride bonds significantly improves the stability. These stable polyanhydrides can provide convenience to use in control drug-delivery applications. The in vitro drug release study using ibuprofen shows that polymers with aromatic units such as PRAP and PHSAP establish sustained release, which presents more than 50 and 40% of ibuprofen over a period of 28 days.


Assuntos
Anidridos , Polianidridos , Anidridos/química , Ésteres/química , Hidroxiácidos , Ibuprofeno , Ácidos Ricinoleicos/química
4.
ACS Biomater Sci Eng ; 8(6): 2500-2507, 2022 06 13.
Artigo em Inglês | MEDLINE | ID: mdl-35604784

RESUMO

Seasonal influenza A virus infections present substantial costs to both health and economic resources each year. Current seasonal influenza vaccines provide suboptimal protection and require annual reformulation to match circulating strains. In this work, a recombinant equine H3N8 hemagglutinin trimer (rH33) known to generate cross-protective antibodies and protect animals against sublethal, heterologous virus challenge was used as a candidate vaccine antigen. Nanoadjuvants such as polyanhydride nanoparticles and pentablock copolymer hydrogels have been shown to be effective adjuvants, inducing both rapid and long-lived protective immunity against influenza A virus. In this work, polyanhydride nanoparticles and pentablock copolymer hydrogels were used to provide sustained release of the novel rH33 while also facilitating the retention of its structure and antigenicity. These studies lay the groundwork for the development of a novel universal influenza A virus nanovaccine by combining the equine H3N8 rH33 and polymeric nanoadjuvant platforms.


Assuntos
Vírus da Influenza A Subtipo H3N8 , Vírus da Influenza A , Nanopartículas , Polianidridos , Animais , Anticorpos Antivirais , Hemaglutininas , Cavalos , Hidrogéis , Nanopartículas/química , Polianidridos/química
5.
J Biomed Mater Res B Appl Biomater ; 110(2): 450-459, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34312984

RESUMO

Parkinson's disease (PD) is a devastating neurodegenerative disease affecting a large proportion of older adults. Exposure to pesticides like rotenone is a leading cause for PD. To reduce disease progression and prolong life expectancy, it is important to target disease mechanisms that contribute to dopaminergic neuronal atrophy, including mitochondrial dysfunction. Achieving targeted mitochondrial delivery is difficult for many therapeutics by themselves, necessitating higher therapeutic doses that could lead to toxicity. To minimize this adverse effect, targeted nano-carriers such as polyanhydride nanoparticles (NPs) can protect therapeutics from degradation and provide sustained release, enabling fewer administrations and lower therapeutic dose. This work expands upon the use of the polyanhydride NP platform for targeted drug delivery by functionalizing the polymer with a derivative of triphenylphosphonium called (3-carboxypropyl) triphenylphosphonium (CPTP) using a novel method that enables longer CPTP persistence on the NPs. The extent to which neurons internalized both nonfunctionalized and functionalized NPs was tested. Next, the efficacy of these nanoformulations in treating rotenone-induced mitochondrial dysfunction in the same cell line was evaluated using a novel neuroprotective drug, mito-metformin. CPTP functionalization significantly improved NP internalization by neuronal cells. This was correlated with significant protection by CPTP-functionalized, mito-metformin encapsulated NPs against rotenone-induced mitochondrial dysfunction. However, nonfunctionalized, mito-metformin encapsulated NPs and soluble mito-metformin administered at the same dose did not significantly protect cells from rotenone-induced toxicity. These results indicate that the targeted NP platform can provide enhanced dose-sparing and potentially reduce the occurrence of systemic side-effects for PD therapeutics.


Assuntos
Nanopartículas , Doenças Neurodegenerativas , Polianidridos , Idoso , Humanos , Mitocôndrias/metabolismo , Doenças Neurodegenerativas/metabolismo , Polianidridos/metabolismo , Polianidridos/farmacologia , Rotenona/metabolismo , Rotenona/toxicidade
6.
J Vis Exp ; (172)2021 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-34251369

RESUMO

Cytokine therapy is a promising immunotherapeutic strategy that can produce robust antitumor immune responses in cancer patients. The proinflammatory cytokine interleukin-1 alpha (IL-1α) has been evaluated as an anticancer agent in several preclinical and clinical studies. However, dose-limiting toxicities, including flu-like symptoms and hypotension, have dampened the enthusiasm for this therapeutic strategy. Polyanhydride nanoparticle (NP)-based delivery of IL-1α would represent an effective approach in this context since this may allow for a slow and controlled release of IL-1α systemically while reducing toxic side effects. Here an analysis of the antitumor activity of IL-1α-loaded polyanhydride NPs in a head and neck squamous cell carcinoma (HNSCC) syngeneic mouse model is described. Murine oropharyngeal epithelial cells stably expressing HPV16 E6/E7 together with hRAS and luciferase (mEERL) cells were injected subcutaneously into the right flank of C57BL/6J mice. Once tumors reached 3-4 mm in any direction, a 1.5% IL-1a - loaded 20:80 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane:1,6-bis(p-carboxyphenoxy)hexane (CPTEG: CPH) nanoparticle (IL-1α-NP) formulation was administered to mice intraperitoneally. Tumor size and body weight were continuously measured until tumor size or weight loss reached euthanasia criteria. Blood samples were taken to evaluate antitumor immune responses by submandibular venipuncture, and inflammatory cytokines were measured through cytokine multiplex assays. Tumor and inguinal lymph nodes were resected and homogenized into a single-cell suspension to analyze various immune cells through multicolor flow cytometry. These standard methods will allow investigators to study the antitumor immune response and potential mechanism of immunostimulatory NPs and other immunotherapy agents for cancer treatment.


Assuntos
Neoplasias de Cabeça e Pescoço , Nanopartículas , Polianidridos , Animais , Humanos , Interleucina-1alfa , Camundongos , Camundongos Endogâmicos C57BL
7.
J Biomater Sci Polym Ed ; 32(10): 1312-1336, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33874849

RESUMO

The present study focuses on the synthesis and characterization of hydroxyapatite-collagen nanoparticles incorporated polyanhydride paste and investigating its bone regeneration capacity in vitro. Photocrosslinkable polyanhydride paste was prepared after synthesizing methacrylate derivatives of 1,6-bis(p-carboxyphenoxy)hexane (MCPH) and sebacic acid dimethacrylate (MSA). These multifunctional monomers, namely 45 wt% MSA, 45 wt% MCPH in addition to 10 wt% poly(ethylene glycol)diacrylate (PEGDA) were photopolymerized under ultraviolet light (365 nm) to produce highly crosslinked polyanhydride networks using camphroquinone (CQ)/ethyl 4-(dimethylamino)benzoate [4-EDMAB] for light initiated crosslinking and benzoyl peroxide (BPO)/dimethyl toludine (DMT) for chemically initiated crosslinking. Separately, using the co-precipitation process, (1 wt%) Si, (1 wt%) Sr, and (0.5 + 0.5) wt% Si/Sr was doped into hydroxyapatite-collagen nanoparticles in size range between 50 and 70 nm. Si, Sr, and both Si/Sr doped hydroxyapatite-collagen nanoparticles to the extent 10 wt% were added to polyanhydride monomer mixture containing 40 wt% MSA, 40 wt% MCPH and 10 wt% PEGDA and subsequently photopolymerized as previously mentioned. Incorporation of hydroxyapatite-collagen nanoparticles to the extent of 10 wt% into polyanhydride matrix enhanced compressive strength of the hardened paste from 30 to 49 MPa. Mesenchymal stem cells obtained from the human umbilical cord were cultured onto pure polyanhydride and hydroxyapatite-collagen added scaffold to assess their cellular proliferation and differentiation capacity to bone cell. MTT assay showed that mesenchymal stem cell proliferation was significantly higher in Si/Sr binary doped hydroxyapatite-collagen-polyanhydride sample as compared to other samples. Again from immunocytochemistry study using confocal images suggested that expression of osteocalcin, a marker indicating differentiation into osteoblast, was the highest in Si/Sr binary doped hydroxyapatite-collagen-polyanhydride sample against the other samples studied in this case. This study thus summarizes the development of photocurable biocomposites containing polyanhydride and Si, Sr doped hydroxyapatite-collagen nanoparticles that exhibited tremendous promise to regenerate bone tissues in complex-shaped musculoskeletal defect sites.


Assuntos
Substitutos Ósseos , Nanopartículas , Polianidridos , Osso e Ossos , Colágeno , Durapatita , Humanos
8.
J Immunol ; 206(9): 2122-2134, 2021 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-33827894

RESUMO

Respiratory syncytial virus (RSV) is a leading cause of lower respiratory tract infection in both young children and in older adults. Despite the morbidity, mortality, and high economic burden caused by RSV worldwide, no licensed vaccine is currently available. We have developed a novel RSV vaccine composed of a prefusion-stabilized variant of the fusion (F) protein (DS-Cav1) and a CpG oligodeoxynucleotide adjuvant encapsulated within polyanhydride nanoparticles, termed RSVNanoVax. A prime-boost intranasal administration of RSVNanoVax in BALB/c mice significantly alleviated weight loss and pulmonary dysfunction in response to an RSV challenge, with protection maintained up to at least 6 mo postvaccination. In addition, vaccinated mice exhibited rapid viral clearance in the lungs as early as 2 d after RSV infection in both inbred and outbred populations. Vaccination induced tissue-resident memory CD4 and CD8 T cells in the lungs, as well as RSV F-directed neutralizing Abs. Based on the robust immune response elicited and the high level of durable protection observed, our prefusion RSV F nanovaccine is a promising new RSV vaccine candidate.


Assuntos
Imunidade Celular/imunologia , Polianidridos/química , Vacinas contra Vírus Sincicial Respiratório/imunologia , Vírus Sincicial Respiratório Humano/imunologia , Animais , Feminino , Camundongos , Camundongos Endogâmicos BALB C
9.
Int J Mol Sci ; 22(3)2021 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-33499242

RESUMO

In the course of this study, a series of novel, biodegradable polyanhydrides based on betulin disuccinate and dicarboxylic derivatives of poly(ethylene glycol) were prepared by two-step polycondensation. These copolymers can be used as carriers in drug delivery systems, in the form of microspheres. Betulin and its derivatives exhibit a broad spectrum of biological activity, including cytotoxic activity, which makes them promising substances for use as therapeutic agents. Microspheres that were prepared from betulin based polyanhydrides show promising properties for use in application in drug delivery systems, including inhalation systems. The obtained copolymers release the active substance-betulin disuccinate-as a result of hydrolysis under physiological conditions. The use of a poly(ethylene glycol) derivative as a co-monomer increases the solubility and bioavailability of the obtained compounds. Microspheres with diameters in the range of 0.5-25 µm were prepared by emulsion solvent evaporation method and their physicochemical and aerodynamic properties were analyzed. The morphological characteristics of the microspheres depended on the presence of poly(ethylene glycol) (PEG) segment within the structure of polyanhydrides. The porosity of the particles depended on the amount and molecular weight of the PEG used and also on the speed of homogenization. The most porous particles were obtained from polyanhydrides containing 20% wt. of PEG 600 by using a homogenization speed of 18,000 rpm.


Assuntos
Sistemas de Liberação de Medicamentos , Compostos Fitoquímicos/química , Polianidridos/química , Polietilenoglicóis/química , Triterpenos/química , Células A549 , Aerossóis , Linhagem Celular Tumoral , Portadores de Fármacos/química , Células HeLa , Células Hep G2 , Humanos , Concentração de Íons de Hidrogênio , Hidrólise , Células MCF-7 , Espectroscopia de Ressonância Magnética , Microesferas , Peso Molecular , Tamanho da Partícula , Polímeros/química , Porosidade , Solventes
10.
J Biomed Mater Res A ; 109(6): 893-902, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-32776461

RESUMO

Pancreatic cancer (PC) is one of the most lethal malignancies and represents an increasing and challenging threat, especially with an aging population. The identification of immunogenic PC-specific upregulated antigens and an enhanced understanding of the immunosuppressive tumor microenvironment have provided opportunities to enable the immune system to recognize cancer cells. Due to its differential upregulation and functional role in PC, the transmembrane mucin MUC4 is an attractive target for immunotherapy. In the current study we characterized the antigen stability, antigenicity and release kinetics of a MUC4ß-nanovaccine to guide further optimization and, in vivo evaluation. Amphiphilic polyanhydride copolymers based on 20 mol % 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane and 80 mol % 1,6-bis(p-carboxyphenoxy)hexane were used to synthesize nanoparticles. Structurally stable MUC4ß protein was released from the particles in a sustained manner and characterized by gel electrophoresis and fluorescence spectroscopy. Modest levels of protein degradation were observed upon release. The released protein was also analyzed by MUC4ß-specific monoclonal antibodies using ELISA and showed no significant loss of epitope availability. Further, mice immunized with multiple formulations of combination vaccines containing MUC4ß-loaded nanoparticles generated MUC4ß-specific antibody responses. These results indicate that polyanhydride nanoparticles are viable MUC4ß vaccine carriers, laying the foundation for evaluation of this platform for PC immunotherapy.


Assuntos
Antígenos de Neoplasias/química , Vacinas Anticâncer/química , Mucina-4/química , Nanopartículas , Neoplasias Pancreáticas/química , Polianidridos/química , Animais , Portadores de Fármacos , Epitopos , Humanos , Camundongos
11.
Int J Nanomedicine ; 15: 6579-6592, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32982219

RESUMO

INTRODUCTION: Adjuvants and immunotherapies designed to activate adaptive immunity to eliminate infectious disease and tumors have become an area of interest aimed at providing a safe and effective strategy to prevent or eliminate disease. Existing approaches would benefit from the development of immunization regimens capable of inducing efficacious cell-mediated immunity directed toward CD8+ T cell-specific antigens. This goal is critically dependent upon appropriate activation of antigen-presenting cells (APCs) most notably dendritic cells (DCs). In this regard, polyanhydride particles have been shown to be effectively internalized by APCs and induce activation. METHODS: Here, a prophylactic vaccine regimen designed as a single-dose polyanhydride nanovaccine encapsulating antigen is evaluated for the induction of CD8+ T cell memory in a model system where antigen-specific protection is restricted to CD8+ T cells. Bone marrow-derived dendritic cells (BMDCs) are used as an in vitro model system to evaluate the magnitude and phenotype of APC activation. Primary DCs, particularly those with described ability to activate CD8+ T cells, are also evaluated for their in vitro responses to polyanhydride nanoparticles. RESULTS: Herein, polyanhydride nanoparticles are shown to induce potent in vitro upregulation of costimulatory molecules on the cell surface of BMDCs. In contrast to the classically used TLR agonists, nanoparticles did not induce large amounts of pro-inflammatory cytokines, did not induce characteristic metabolic response of DCs, nor produce innate antimicrobial effector molecules, such as nitric oxide (NO). The polyanhydride nanovaccine results in protective CD8+ T cell responses as measured by inhibition of tumor progression and survival. DISCUSSION: Together, these results suggest that the use of a polyanhydride-based nanovaccine can be an effective approach to inducing antigen-specific CD8+ T cell memory by providing antigen delivery and DC activation while avoiding overt inflammatory responses typically associated with traditional adjuvants.


Assuntos
Linfócitos T CD8-Positivos/efeitos dos fármacos , Vacinas Anticâncer/imunologia , Células Dendríticas/efeitos dos fármacos , Nanopartículas/administração & dosagem , Polianidridos/farmacologia , Adjuvantes Imunológicos , Animais , Apresentação de Antígeno/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Vacinas Anticâncer/administração & dosagem , Vacinas Anticâncer/farmacologia , Citocinas/metabolismo , Células Dendríticas/imunologia , Feminino , Imunidade Celular/efeitos dos fármacos , Memória Imunológica/efeitos dos fármacos , Imunoterapia/métodos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Nanopartículas/química , Neoplasias Experimentais/imunologia , Neoplasias Experimentais/terapia , Polianidridos/administração & dosagem
12.
PLoS Negl Trop Dis ; 14(9): e0008365, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32898130

RESUMO

Insecticide resistance poses a significant threat to the control of arthropods that transmit disease agents. Nanoparticle carriers offer exciting opportunities to expand the armamentarium of insecticides available for public health and other pests. Most chemical insecticides are delivered by contact or feeding, and from there must penetrate various biological membranes to reach target organs and kill the pest organism. Nanoparticles have been shown to improve bioactive compound navigation of such barriers in vertebrates, but have not been well-explored in arthropods. In this study, we explored the potential of polyanhydride micro- and nanoparticles (250 nm- 3 µm), labeled with rhodamine B to associate with and/or transit across insect biological barriers, including the cuticle, epithelium, midgut and ovaries, in female Ae. aeygpti mosquitoes. Mosquitoes were exposed using conditions to mimic surface contact with a residual spray or paint, topical exposure to mimic contact with aerosolized insecticide, or per os in a sugar meal. In surface contact experiments, microparticles were sometimes observed in association with the exterior of the insect cuticle. Nanoparticles were more uniformly distributed across exterior tissues and present at higher concentrations. Furthermore, by surface contact, topical exposure, or per os, particles were detected in internal organs. In every experiment, amphiphilic polyanhydride nanoparticles associated with internal tissues to a higher degree than hydrophobic nanoparticles. In vitro, nanoparticles associated with Aedes aegypti Aag2 cells within two hours of exposure, and particles were evident in the cytoplasm. Further studies demonstrated that particle uptake is dependent on caveolae-mediated endocytosis. The propensity of these nanoparticles to cross biological barriers including the cuticle, to localize in target tissue sites of interest, and to reach the cytoplasm of cells, provides great promise for targeted delivery of insecticidal candidates that cannot otherwise reach these cellular and subcellular locations.


Assuntos
Aedes/fisiologia , Nanopartículas , Polianidridos , Aedes/citologia , Animais , Linhagem Celular , Endocitose , Feminino , Controle de Mosquitos/métodos , Rodaminas/química , Distribuição Tecidual
13.
ACS Comb Sci ; 22(4): 172-183, 2020 04 13.
Artigo em Inglês | MEDLINE | ID: mdl-32125826

RESUMO

Combinatorial techniques can accelerate the discovery and development of polymeric nanodelivery devices by pairing high-throughput synthesis with rapid materials characterization. Biodegradable polyanhydrides demonstrate tunable release, high cellular internalization, and dose sparing properties when used as nanodelivery devices. This nanoparticle platform shows promising potential for small molecule drug delivery, but the pace of understanding and rational design of these nanomedicines is limited by the low throughput of conventional characterization. This study reports the use of a high-throughput method to synthesize libraries of a newly synthesized, rapidly eroding polyanhydride copolymer based on 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG) and sebacic acid (SA) monomers. The high-throughput method enabled efficient screening of copolymer microstructure, revealing weak block-type and alternating architectures. The high-throughput method was adapted to synthesize nanoparticle libraries encapsulating hydrophobic model drugs. Drug release from these nanoparticles was rapid, with a majority of the payload released within 3 days. Drug release was dramatically slowed at acidic pH, which could be useful for oral drug delivery. Rhodamine B (RhoB) release kinetics generally followed patterns of polymer erosion kinetics, while Coomassie brilliant blue (CBB) released the fastest from the slowest degrading polymer chemistry and vice versa. These differences in trends between copolymer chemistry and release kinetics were hypothesized to arise from differences in mixing thermodynamics. A high-throughput method was developed to synthesize polymer-drug film libraries and characterize mixing thermodynamics by melting point depression. Rhodamine B had a negative χ for all copolymers with <30 mol % CPTEG tested, indicating a tendency toward miscibility. By contrast, CBB χ increased, eventually becoming positive near 15:85 CPTEG:SA, with increasing CPTEG content. This indicates an increasing tendency toward phase separation in CPTEG-rich copolymers. These in vitro results screening polymer-drug interactions showed good agreement with in silico predictions from Hansen solubility parameter estimation and were able to explain the observed differences in model drug release trends.


Assuntos
Técnicas de Química Combinatória , Ensaios de Triagem em Larga Escala , Nanopartículas/química , Polianidridos/química , Polianidridos/síntese química , Benzenossulfonatos/química , Liberação Controlada de Fármacos , Cinética , Tamanho da Partícula , Rodaminas/química , Propriedades de Superfície
14.
ACS Biomater Sci Eng ; 6(1): 265-276, 2020 01 13.
Artigo em Inglês | MEDLINE | ID: mdl-33463223

RESUMO

There is a currently a need to develop adjuvants that are best suited to simultaneously enhance immune responses, induce immunologic memory, improve patient compliance (i.e., reduce doses and inflammation), and provide vaccine shelf stability for stockpiling and global deployment to challenging environments. Biodegradable polyanhydrides have been investigated extensively to overcome such challenges. It has been shown that controlling copolymer composition can result in chemistry-dependent immunomodulatory capabilities. These studies have revealed that copolymers rich in sebacic acid (SA) are highly internalized by antigen presenting cells and confer improved shelf stability of encapsulated proteins, while copolymers rich in 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG) also exhibit enhanced internalization by and activation of antigen presenting cells (APCs), in addition to providing superior retention of protein stability following encapsulation and release. However, to date, CPTEG:SA copolymers have not been synthesized and described. In this work, we hypothesized that new copolymers composed of CPTEG and SA would combine the advantages of both monomers in terms of enhanced thermal properties, maintaining antigenicity of encapsulated proteins following nanoparticle synthesis, and superior cellular internalization and activation by APCs, demonstrated by the upregulation of costimulatory markers CD80, CD86, and CD40, as well as the secretion of proinflammatory cytokines IL-6, IL-1ß, and TNF-α. Herein, we describe the synthesis and design of novel CPTEG:SA nanoparticles with improved thermal properties, payload stability, and internalization by antigen presenting cells for applications in vaccine delivery. The performance of these new CPTEG:SA formulations was compared to that of traditional polyanhydride copolymers.


Assuntos
Adjuvantes Imunológicos , Nanopartículas , Polianidridos , Vacinas , Adjuvantes Imunológicos/farmacologia , Sistemas de Liberação de Medicamentos , Humanos
15.
Vet Microbiol ; 237: 108401, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31585639

RESUMO

Influenza results in significant economic loss in the swine industry each year. A broadly protective swine influenza vaccine would have the dual benefit of protecting pigs from influenza A viruses (IAVs) and limiting their possible zoonotic transmission to humans. In this study, we developed polyanhydride nanoparticles-based swine influenza vaccine (KAg + CpG-nanovaccine) co-encapsulating inacticated/killed soluble antigen (KAg) and Toll-like receptor (TLR)-9 agonist (CpG-ODN). The immunogenicity and protective efficacy of KAg + CpG-nanovaccine was compared with KAg vaccine containing five-times greater quantity of antigens following heterologous virus challenge. Prime-boost intranasally delivered KAg + CpG-nanovaccine induced significantly higher levels of cross-reactive antigen-specific IgA antibody responses in the nasal cavity, greater lymphoproliferative response in peripheral blood mononuclear cells (PBMCs), and higher IFN-γ secretion during antigen-induced recall responses of PBMCs and tracheobronchial lymph nodes cells compared to those immunized with KAg alone. Importantly, KAg + CpG-nanovaccine provided better protective efficacy through a significant reduction in influenza-induced fever, 16-fold reduction of nasal virus shedding and 80-fold reduction in lung virus titers compared to those immunized with soluble KAg. Our results indicated that CpG-ODN-adjuvanted polyanhydride nanovaccine can induce higher mucosal antibody and cellular immune responses in pigs; and provide better protection as compared with intranasally delivered soluble KAg.


Assuntos
Vacinas contra Influenza/imunologia , Oligodesoxirribonucleotídeos/farmacologia , Infecções por Orthomyxoviridae/veterinária , Doenças dos Suínos/prevenção & controle , Adjuvantes Imunológicos , Administração Intranasal , Animais , Anticorpos Antivirais , Antígenos Virais/imunologia , Feminino , Imunidade nas Mucosas , Imunoglobulina A/imunologia , Interferon gama/metabolismo , Leucócitos Mononucleares , Masculino , Nanoestruturas , Oligodesoxirribonucleotídeos/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Polianidridos , Suínos , Vacinas de Produtos Inativados/imunologia
16.
Nanomedicine ; 21: 102055, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31319179

RESUMO

The primary objective of this study was to enhance the antitumor efficacy of a model cancer vaccine through co-delivery of pentaerythritol lipid A (PELA), an immunological adjuvant, and a model tumor antigen, ovalbumin (OVA), separately loaded into polyanhydride particles (PA). In vitro experiments showed that encapsulation of PELA into PA (PA-PELA) significantly enhanced its stimulatory capacity on dendritic cells as evidenced by increased levels of the cell surface costimulatory molecules, CD80/CD86. In vivo experiments showed that PA-PELA, in combination with OVA-loaded PA (PA-OVA), significantly expanded the OVA-specific CD8+ T lymphocyte population compared to PA-OVA alone. Furthermore, OVA-specific serum antibody titers of mice vaccinated with PA-OVA/PA-PELA displayed a significantly stronger shift toward a Th1-biased immune response compared to PA-OVA alone, as evidenced by the substantially higher IgG2C:IgG1 ratios achieved by the former. Analysis of E.G7-OVA tumor growth curves showed that mice vaccinated with PA-OVA/PA-PELA had the slowest average tumor growth rate.


Assuntos
Vacinas Anticâncer/farmacologia , Células Dendríticas/efeitos dos fármacos , Imunidade Celular/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/farmacologia , Animais , Antígeno B7-1/imunologia , Antígeno B7-2/imunologia , Linfócitos T CD8-Positivos , Proliferação de Células/efeitos dos fármacos , Células Dendríticas/imunologia , Humanos , Imunidade Celular/imunologia , Imunoglobulina G/imunologia , Lipídeo A/química , Lipídeo A/farmacologia , Camundongos , Neoplasias/imunologia , Neoplasias/patologia , Polianidridos/química , Polianidridos/farmacologia , Propilenoglicóis/química , Propilenoglicóis/farmacologia , Receptores de IgG/imunologia , Células Th1/efeitos dos fármacos , Células Th1/imunologia
17.
Drug Dev Ind Pharm ; 45(10): 1635-1645, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31342792

RESUMO

Objective: The aim of the study was to formulate, cyclodextrin (CD)-polyanhydride (PA) nanoparticles (CPNs) with rosuvastatin calcium (RCa) in order to enhance the poor oral bioavailability. Methods: CPNs containing RCa/CD complexes were prepared by a modified solvent displacement method and morphological analyses, particle size (PS), polydispersity index (PDI), zeta potential (ZP), encapsulation efficiency (EE), DSC, FT-IR, XRD, 1H-NMR analyses were performed. In vitro release properties, release kinetics, cytotoxicity, in vitro permeability and pharmacokinetic studies were also studied. The stability of the formulations were evaluated during the storage period of 3 months. Results: The physicochemical studies showed that the RCa/CD complexes were well incorporated into CPNs resulted in nanosized particles (215.22 and 189.13 nm) with homogenous size distribution (PDI: 0.203 and 0.182) with relatively high incorporation capacity (76.11 and 68.18%) for the CPN1 and CPN2 formulations respectively. Sustained release of RCa from CPNs were achieved. The cytotoxicity values showed that the safety of the formulations. According to permeability studies, pure RCa had lowest permeability data (3.08 × 10-7 cm⋅s-1 Papp value) while CPNs gained higher permeability data (1.36 × 10-5 and 1.12 × 10-5 cm⋅s-1 Papp values) for the CPN1 and CPN2 formulations respectively. CPN2 formulation was selected for pharmacokinetic studies and analyses results demonstrated that approximately 8-fold relative oral bioavailability enhancement compared to the pure RCa was achieved. Conclusion: Considering the analyses results of the study, CPNs can be regarded as suitable, safe, functional oral delivery systems for RCa with enhanced oral bioavailability.


Assuntos
Ciclodextrinas/química , Ciclodextrinas/farmacocinética , Nanopartículas/química , Polianidridos/química , Polianidridos/farmacocinética , Rosuvastatina Cálcica/química , Rosuvastatina Cálcica/farmacocinética , Animais , Disponibilidade Biológica , Células CACO-2 , Linhagem Celular Tumoral , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacocinética , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Humanos , Masculino , Tamanho da Partícula , Permeabilidade/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley
18.
Mol Pharm ; 16(5): 1917-1928, 2019 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-30973741

RESUMO

Drug delivery vehicles can improve the functional efficacy of existing antimicrobial therapies by improving biodistribution and targeting. A critical property of such nanomedicine formulations is their ability to control the release kinetics of their payloads. The combination of (and interactions among) polymer, drug, and nanoparticle properties gives rise to nonlinear behavioral relationships and large data space. These factors complicate both first-principles modeling and screening of nanomedicine formulations. Predictive analytics may offer a more efficient approach toward the rational design of nanomedicines by identifying key descriptors and correlating them to nanoparticle release behavior. In this work, antibiotic release kinetics data were generated from polyanhydride nanoparticle formulations with varying copolymer compositions, encapsulated drug type, and drug loading. Four antibiotics, doxycycline, rifampicin, chloramphenicol, and pyrazinamide, were used. Linear manifold learning methods were used to relate drug release properties with polymer, drug, and nanoparticle properties, and key descriptors were identified that are highly correlated with release properties. However, these linear methods could not predict release behavior. Nonlinear multivariate modeling based on graph theory was then used to deconvolute the governing relationships between these properties, and predictive models were generated to rapidly screen lead nanomedicine formulations with desirable release properties with minimal nanoparticle characterization. Release kinetics predictions of two drugs containing atoms not included in the model showed good agreement with experimental results, validating the model and indicating its potential to virtually explore new polymer and drug pairs not included in the training data set. The models were shown to be robust after the inclusion of these new formulations, in that the new inclusions did not significantly change model regression. This approach provides the first step toward the development of a framework that can be used to rationally design nanomedicine formulations by selecting the appropriate carrier for a drug payload to program desirable release kinetics.


Assuntos
Ciência de Dados/métodos , Desenho de Fármacos , Liberação Controlada de Fármacos , Modelos Biológicos , Nanopartículas/química , Antibacterianos/farmacocinética , Antibacterianos/uso terapêutico , Infecções Bacterianas/tratamento farmacológico , Bases de Dados de Produtos Farmacêuticos , Composição de Medicamentos/métodos , Sistemas de Liberação de Medicamentos , Humanos , Nanomedicina , Polianidridos/química , Polímeros/química , Distribuição Tecidual
19.
Biomater Sci ; 7(3): 809-821, 2019 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-30663733

RESUMO

Immunosenescence poses a formidable challenge in designing effective influenza vaccines for aging populations. While approved vaccines against influenza viruses exist, their efficacy in older adults is significantly decreased due to the diminished capabilities of innate and adaptive immune responses. In this work, the ability of a combination nanovaccine containing both recombinant hemagglutinin and nucleoprotein to provide protection against seasonal influenza virus infection was examined in young and aged mice. Vaccine formulations combining two nanoadjuvants, polyanhydride nanoparticles and pentablock copolymer micelles, were shown to enhance protection against challenge compared to each adjuvant alone in young mice. Nanoparticles were shown to enhance in vitro activation of dendritic cells isolated from aged mice, while both nanoadjuvants did not induce proinflammatory cytokine secretion which may be detrimental in aged individuals. In addition, the combination nanovaccine platform was shown to induce demonstrable antibody titers in both young and aged mice that correlated with the maintenance of body weight post-challenge. Collectively, these data demonstrate that the combination nanovaccine platform is a promising technology for influenza vaccines for older adults.


Assuntos
Envelhecimento , Vacinas contra Influenza/imunologia , Nanopartículas/química , Infecções por Orthomyxoviridae/prevenção & controle , Adjuvantes Imunológicos/química , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Células Dendríticas/citologia , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Feminino , Vírus da Influenza A/patogenicidade , Vacinas contra Influenza/química , Pulmão/virologia , Camundongos , Camundongos Endogâmicos BALB C , Micelas , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/mortalidade , Polianidridos/química , Polímeros/química , Taxa de Sobrevida
20.
J Pharmacol Exp Ther ; 370(3): 855-863, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30361239

RESUMO

Many factors affect vaccine efficacy. One of the most salient is the frequency and intervals of vaccine administration. In this study, we assessed the vaccine administration modality for a recently reported polyanhydride-based vaccine formulation, shown to generate antitumor activity. Polyanhydride particles encapsulating ovalbumin (OVA) were prepared using a double-emulsion technique and subcutaneously delivered to mice either as a single-dose or as prime-boost vaccine regimens in which two different time intervals between prime and boost were assessed (7 or 21 days). This was followed by measurement of cellular and humoral immune responses, and subsequent challenge of the mice with a lethal dose of E.G7-OVA cells to evaluate tumor protection. Interestingly, a single dose of the polyanhydride particle-based formulation induced sustained OVA-specific cellular immune responses just as effectively as the prime-boost regimens. In addition, mice receiving single-dose vaccine had similar levels of protection against tumor challenge compared with mice administered prime-boosts. In contrast, measurements of OVA-specific IgG antibody titers indicated that a booster dose was required to stimulate strong humoral immune responses, since it was observed that mice administered a prime-boost vaccine had significantly higher OVA-specific IgG1 serum titers than mice administered a single dose. These findings indicate that the requirement for a booster dose using these particles appears unnecessary for the generation of effective cellular immunity.


Assuntos
Vacinas Anticâncer/administração & dosagem , Polianidridos , Animais , Composição de Medicamentos , Excipientes , Feminino , Imunidade Celular , Imunidade Humoral , Imunização Secundária , Imunoglobulina G/análise , Camundongos , Camundongos Endogâmicos C57BL , Nanopartículas , Neoplasias Experimentais/induzido quimicamente , Neoplasias Experimentais/prevenção & controle , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Suspensões
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