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1.
Eur J Pharm Biopharm ; 163: 198-211, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33852968

RESUMO

Oral inhalation is the preferred route for delivery of small molecules to the lungs, because high tissue levels can be achieved shortly after application. Biologics are mainly administered by intravenous injection but inhalation might be beneficial for the treatment of lung diseases (e.g. asthma). This review discusses biological and pharmaceutical challenges for delivery of biologics and describes promising candidates. Insufficient stability of the proteins during aerosolization and the biological environment of the lung are the main obstacles for pulmonary delivery of biologics. Novel nebulizers will improve delivery by inducing less shear stress and administration as dry powder appears suitable for delivery of biologics. Other promising strategies include pegylation and development of antibody fragments, while carrier-encapsulated systems currently play no major role in pulmonary delivery of biologics for lung disease. While development of various biologics has been halted or has shown little effects, AIR DNase, alpha1-proteinase inhibitor, recombinant neuraminidase, and heparin are currently being evaluated in phase III trials. Several biologics are being tested for the treatment of coronavirus disease (COVID)-19, and it is expected that these trials will lead to improvements in pulmonary delivery of biologics.


Assuntos
Pneumopatias/tratamento farmacológico , Pulmão/efeitos dos fármacos , Peptídeos/administração & dosagem , Proteínas/administração & dosagem , Administração por Inalação , Administração Oral , Animais , Produtos Biológicos/administração & dosagem , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Humanos , Nebulizadores e Vaporizadores , Pós/administração & dosagem
2.
Nutrients ; 13(2)2021 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-33530344

RESUMO

Hepatic encephalopathy (HE) is a common neurological consequence in patients with cirrhosis and has a healthcare burden of USD 5370 to 50,120 per patient annually. HE significantly hampers the quality of life and is a major cause of morbidity and mortality. Patients with cirrhosis are at a high risk for protein-calorie malnutrition due to altered metabolism. Current evidence has changed the old belief of protein restriction in patients with cirrhosis and now 1.2 to 1.5 g/kg/day protein intake is recommended. Case series and studies with small numbers of participants showed that a vegetarian protein diet decreases the symptoms of HE when compared to a meat-based diet, but the evidence is limited and requires further larger randomized controlled trials. However, vegetable or milk-based protein diets are good substitutes for patients averse to meat intake. Branch chain amino acids (BCAA) (leucine, isoleucine and valine) have also been shown to be effective in alleviating symptoms of HE and are recommended as an alternative therapy in patients with cirrhosis for the treatment of HE. In this review, we provide an overview of current literature evaluating the role of protein intake in the management of HE in cirrhosis.


Assuntos
Dieta Vegetariana , Proteínas na Dieta , Encefalopatia Hepática , Carne , Proteínas/administração & dosagem , Aminoácidos de Cadeia Ramificada , Animais , Bases de Dados Factuais , Fibrose , Encefalopatia Hepática/terapia , Humanos , Proteínas de Vegetais Comestíveis , Desnutrição Proteico-Calórica , Qualidade de Vida , Vegetarianos
3.
J Clin Pharmacol ; 60(10): 1275-1293, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32779201

RESUMO

The coronavirus disease 2019 (COVID-19) pandemic caused by infection with SARS-CoV-2 has led to more than 600 000 deaths worldwide. Patients with severe disease often experience acute respiratory distress characterized by upregulation of multiple cytokines. Immunomodulatory biological therapies are being evaluated in clinical trials for the management of the systemic inflammatory response and pulmonary complications in patients with advanced stages of COVID-19. In this review, we summarize the clinical pharmacology considerations in the development of immunomodulatory therapeutic proteins for mitigating the heightened inflammatory response identified in COVID-19.


Assuntos
Infecções por Coronavirus/tratamento farmacológico , Fatores Imunológicos/administração & dosagem , Pneumonia Viral/tratamento farmacológico , Proteínas/administração & dosagem , Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/imunologia , Desenvolvimento de Medicamentos , Humanos , Fatores Imunológicos/farmacologia , Pandemias , Pneumonia Viral/epidemiologia , Pneumonia Viral/imunologia , Proteínas/imunologia , Proteínas/farmacologia
4.
Adv Exp Med Biol ; 1250: 35-48, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32601936

RESUMO

Injectable in situ-forming hydrogels have been used clinically in diverse biomedical applications. These hydrogels have distinct advantages such as easy management and minimal invasiveness. The hydrogels are aqueous formulations, and a simple injection at the target site replaces a traditional surgical procedure. Here, we review injectable in situ-forming hydrogels that are formulated by physical and chemical methods to deliver proteins and peptides. Prospects for using in situ-forming hydrogels for several specific applications are also discussed.


Assuntos
Hidrogéis , Peptídeos , Proteínas , Sistemas de Liberação de Medicamentos , Humanos , Hidrogéis/administração & dosagem , Hidrogéis/química , Injeções , Peptídeos/administração & dosagem , Proteínas/administração & dosagem
5.
J Pediatr ; 223: 57-63.e5, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32389719

RESUMO

OBJECTIVE: To assess the effect of early life nutrition on structural brain development in 2 cohorts of extremely preterm infants, before and after the implementation of a nutrition regimen containing more protein and lipid. STUDY DESIGN: We included 178 infants retrospectively (median gestational age, 26.6 weeks; IQR, 25.9-27.3), of whom 99 received the old nutrition regimen (cohort A, 2011-2013) and 79 the new nutrition regimen (cohort B, 2013-2015). Intake of protein, lipids, and calories was calculated for the first 28 postnatal days. Brain magnetic resonance imaging (MRI) was performed at 30 weeks postmenstrual age (IQR, 30.3-31.4) and term-equivalent age (IQR, 40.9-41.4). Volumes of 42 (left + right) brain structures were calculated. RESULTS: Mean protein and caloric intake in cohort B (3.4 g/kg per day [P < .001] and 109 kcal/kg per day [P = .038]) was higher than in cohort A (2.7 g/kg per day; 104 kcal/kg per day). At 30 weeks, 22 regions were significantly larger in cohort B compared with cohort A, whereas at term-equivalent age, only the caudate nucleus was significantly larger in cohort B compared with cohort A. CONCLUSIONS: An optimized nutrition protocol in the first 28 days of life is associated with temporarily improved early life brain volumes.


Assuntos
Encéfalo/crescimento & desenvolvimento , Ingestão de Energia , Fenômenos Fisiológicos da Nutrição do Lactente , Lactente Extremamente Prematuro/crescimento & desenvolvimento , Encéfalo/diagnóstico por imagem , Estudos Controlados Antes e Depois , Feminino , Humanos , Recém-Nascido , Lipídeos/administração & dosagem , Imagem por Ressonância Magnética , Masculino , Proteínas/administração & dosagem , Estudos Retrospectivos
6.
Nature ; 579(7799): 421-426, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32188939

RESUMO

Bioorthogonal chemistry capable of operating in live animals is needed to investigate biological processes such as cell death and immunity. Recent studies have identified a gasdermin family of pore-forming proteins that executes inflammasome-dependent and -independent pyroptosis1-5. Pyroptosis is proinflammatory, but its effect on antitumour immunity is unknown. Here we establish a bioorthogonal chemical system, in which a cancer-imaging probe phenylalanine trifluoroborate (Phe-BF3) that can enter cells desilylates and 'cleaves' a designed linker that contains a silyl ether. This system enabled the controlled release of a drug from an antibody-drug conjugate in mice. When combined with nanoparticle-mediated delivery, desilylation catalysed by Phe-BF3 could release a client protein-including an active gasdermin-from a nanoparticle conjugate, selectively into tumour cells in mice. We applied this bioorthogonal system to gasdermin, which revealed that pyroptosis of less than 15% of tumour cells was sufficient to clear the entire 4T1 mammary tumour graft. The tumour regression was absent in immune-deficient mice or upon T cell depletion, and was correlated with augmented antitumour immune responses. The injection of a reduced, ineffective dose of nanoparticle-conjugated gasdermin along with Phe-BF3 sensitized 4T1 tumours to anti-PD1 therapy. Our bioorthogonal system based on Phe-BF3 desilylation is therefore a powerful tool for chemical biology; our application of this system suggests that pyroptosis-induced inflammation triggers robust antitumour immunity and can synergize with checkpoint blockade.


Assuntos
Preparações de Ação Retardada/administração & dosagem , Neoplasias Mamárias Experimentais/imunologia , Piroptose/imunologia , Animais , Cumarínicos/administração & dosagem , Cumarínicos/química , Cumarínicos/metabolismo , Cumarínicos/farmacocinética , Preparações de Ação Retardada/química , Preparações de Ação Retardada/metabolismo , Preparações de Ação Retardada/farmacocinética , Feminino , Proteínas de Fluorescência Verde/administração & dosagem , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/metabolismo , Proteínas de Fluorescência Verde/farmacocinética , Células HeLa , Humanos , Imunoconjugados/administração & dosagem , Imunoconjugados/química , Imunoconjugados/metabolismo , Imunoconjugados/farmacocinética , Inflamassomos/imunologia , Inflamação/imunologia , Inflamação/metabolismo , Inflamação/patologia , Neoplasias Mamárias Experimentais/metabolismo , Neoplasias Mamárias Experimentais/patologia , Camundongos , Camundongos Endogâmicos BALB C , Oligopeptídeos/administração & dosagem , Oligopeptídeos/química , Oligopeptídeos/metabolismo , Oligopeptídeos/farmacocinética , Receptor de Morte Celular Programada 1/antagonistas & inibidores , Proteínas/administração & dosagem , Proteínas/química , Proteínas/metabolismo , Proteínas/farmacocinética , Silanos/administração & dosagem , Silanos/química , Silanos/metabolismo , Silanos/farmacocinética , Linfócitos T/imunologia , Trastuzumab/administração & dosagem , Trastuzumab/química , Trastuzumab/metabolismo , Trastuzumab/farmacocinética , Ensaios Antitumorais Modelo de Xenoenxerto
7.
PLoS Pathog ; 16(3): e1008339, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32163523

RESUMO

Despite the success of antiretroviral therapy (ART) to halt viral replication and slow disease progression, this treatment is not curative and there remains an urgent need to develop approaches to clear the latent HIV reservoir. The human IL-15 superagonist N-803 (formerly ALT-803) is a promising anti-cancer biologic with potent immunostimulatory properties that has been extended into the field of HIV as a potential "shock and kill" therapeutic for HIV cure. However, the ability of N-803 to reactivate latent virus and modulate anti-viral immunity in vivo under the cover of ART remains undefined. Here, we show that in ART-suppressed, simian-human immunodeficiency virus (SHIV)SF162P3-infected rhesus macaques, subcutaneous administration of N-803 activates and mobilizes both NK cells and SHIV-specific CD8+ T cells from the peripheral blood to lymph node B cell follicles, a sanctuary site for latent virus that normally excludes such effector cells. We observed minimal activation of memory CD4+ T cells and no increase in viral RNA content in lymph node resident CD4+ T cells post N-803 administration. Accordingly, we found no difference in the number or magnitude of plasma viremia timepoints between treated and untreated animals during the N-803 administration period, and no difference in the size of the viral DNA cell-associated reservoir post N-803 treatment. These results substantiate N-803 as a potent immunotherapeutic candidate capable of activating and directing effector CD8+ T and NK cells to the B cell follicle during full ART suppression, and suggest N-803 must be paired with a bona fide latency reversing agent in vivo to facilitate immune-mediated modulation of the latent viral reservoir.


Assuntos
Antirretrovirais/administração & dosagem , Linfócitos B/efeitos dos fármacos , Linfócitos T CD8-Positivos/efeitos dos fármacos , Infecções por HIV/tratamento farmacológico , Interleucina-15/antagonistas & inibidores , Células Matadoras Naturais/efeitos dos fármacos , Proteínas/administração & dosagem , Animais , Linfócitos B/citologia , Linfócitos B/imunologia , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/imunologia , Movimento Celular/efeitos dos fármacos , Modelos Animais de Doenças , Infecções por HIV/genética , Infecções por HIV/imunologia , Infecções por HIV/fisiopatologia , HIV-1/efeitos dos fármacos , HIV-1/fisiologia , Humanos , Interleucina-15/genética , Interleucina-15/imunologia , Células Matadoras Naturais/citologia , Células Matadoras Naturais/imunologia , Linfonodos/efeitos dos fármacos , Linfonodos/imunologia , Macaca mulatta , Proteínas Recombinantes de Fusão , Síndrome de Imunodeficiência Adquirida dos Símios/tratamento farmacológico , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/fisiopatologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/efeitos dos fármacos , Vírus da Imunodeficiência Símia/fisiologia , Latência Viral/efeitos dos fármacos
8.
Annu Rev Pharmacol Toxicol ; 60: 391-415, 2020 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-31914898

RESUMO

The concept of engineering robust protein scaffolds for novel binding functions emerged 20 years ago, one decade after the advent of recombinant antibody technology. Early examples were the Affibody, Monobody (Adnectin), and Anticalin proteins, which were derived from fragments of streptococcal protein A, from the tenth type III domain of human fibronectin, and from natural lipocalin proteins, respectively. Since then, this concept has expanded considerably, including many other protein templates. In fact, engineered protein scaffolds with useful binding specificities, mostly directed against targets of biomedical relevance, constitute an area of active research today, which has yielded versatile reagents as laboratory tools. However, despite strong interest from basic science, only a handful of those protein scaffolds have undergone biopharmaceutical development up to the clinical stage. This includes the abovementioned pioneering examples as well as designed ankyrin repeat proteins (DARPins). Here we review the current state and clinical validation of these next-generation therapeutics.


Assuntos
Descoberta de Drogas/métodos , Engenharia de Proteínas/métodos , Proteínas/administração & dosagem , Animais , Repetição de Anquirina , Humanos , Ligação Proteica , Proteínas/metabolismo , Proteínas/farmacologia
9.
AAPS PharmSciTech ; 21(3): 78, 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31970547

RESUMO

Protein drugs were considered to be the first choice to treat many human diseases, but their clinical application was usually limited by their short half-life and lack of validated targeted therapy. Here, a series of folate-functionalized poly(ethylene glycol)-b-(poly(2-aminoethyl-L-glutamate)-g-poly(L-glutamic acid))s (FA-PEG-b-(PELG-g-PLGA)s) were designed as tumor-targeted carriers for cationic protein delivery. Compared with traditional copolymers consisting of PEG and linear charged hydrophilic blocks, FA-PEG-b-(PELG-g-PLGA) with brush-like polyelectrolyte segments were beneficial to improving their electrostatic interactions with loading protein molecules, thus increasing drug-loading stability and protecting encapsulated proteins from degradation. The designed polymer brushes could efficiently encapsulate cytochrome C (CytC), a cationic model protein, to form polyion complex (PIC) micelles with an average particle size of approximately 200 nm. An in vitro drug release study showed that the drug-loading stability of the formed PIC micelles was largely improved. The functionalization of the block copolymer carriers with a targeting folate group enhanced the tumor cell growth inhibition and total apoptotic rates induced by CytC. Our results shed light on the unique advantages of brush-like polymer carriers in delivering cationic proteins, and the poly(L-glutamic acid)-based linear-brush diblock copolymers could be applied as a versatile delivery platform for molecular targeting in cancer therapy.


Assuntos
Sistemas de Liberação de Medicamentos/métodos , Ácido Glutâmico/síntese química , Poliésteres/síntese química , Polietilenoglicóis/síntese química , Proteínas/síntese química , Animais , Cátions , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Relação Dose-Resposta a Droga , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/síntese química , Portadores de Fármacos/metabolismo , Liberação Controlada de Fármacos , Ácido Glutâmico/administração & dosagem , Ácido Glutâmico/metabolismo , Células HeLa , Humanos , Interações Hidrofóbicas e Hidrofílicas , Camundongos , Células NIH 3T3 , Tamanho da Partícula , Poliésteres/administração & dosagem , Poliésteres/metabolismo , Polietilenoglicóis/administração & dosagem , Polietilenoglicóis/metabolismo , Polímeros/administração & dosagem , Polímeros/síntese química , Polímeros/metabolismo , Proteínas/administração & dosagem , Proteínas/metabolismo
10.
Neurobiol Aging ; 86: 81-91, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31837910

RESUMO

A promising intervention for Alzheimer's disease (AD) would ideally target key pathological factors that are involved in AD pathogenesis. Soluble factors produced by engrafted mesenchymal stem cells (MSCs) mediate potential therapeutic effects in AD. However, these therapeutic benefits are largely hampered by the limited paracrine capacity of MSCs. In this study, we used adenovirus-mediated gene transduction of bone marrow MSCs to deliver exogenous proteins into the brain of APPswe/PSEN1dE9 (APP/PS1) mice in the early stage of impairment. We observed that engrafted MSCs carrying exogenous (C-X3-C motif) ligand 1 (CX3CL1) alone reduced the production of the inflammatory cytokine TNF-ɑ and improved synapse-related protein expression but not cognitive function. Transplantation of MSCs carrying CX3CL1 and Wnt3a (CX3CL1-Wnt3a-MSC) significantly attenuated the learning and memory impairment when compared with a control group. The improvement of neurobehavioral functions in APP/PS1 mice treated with CX3CL1-Wnt3a-MSC was related to the inhibition of microglial neurotoxicity and promotion of hippocampal neurogenesis. Transplantation of CX3CL1-Wnt3a-MSC also regulated phosphoinositide 3-kinase/activated protein kinase B (PI3K/AKT) signaling to inhibit the activity of glycogen synthase kinase 3 beta (GSK3ß). Taken together, these results indicate that the delivery of exogenous proteins via MSCs can modulate microglial function and enhance neurogenesis, thereby providing new insights into AD intervention.


Assuntos
Doença de Alzheimer/terapia , Quimiocina CX3CL1/administração & dosagem , Transplante de Células-Tronco Mesenquimais , Proteínas/administração & dosagem , Proteína Wnt3A/administração & dosagem , Proteína Wnt3A/metabolismo , Adenoviridae , Doença de Alzheimer/metabolismo , Doença de Alzheimer/fisiopatologia , Animais , Células da Medula Óssea , Quimiocina CX3CL1/metabolismo , Cognição , Modelos Animais de Doenças , Células-Tronco Mesenquimais/metabolismo , Camundongos Transgênicos , Neurogênese , Comunicação Parácrina , Transdução Genética , Fator de Necrose Tumoral alfa/metabolismo
11.
Mol Pharm ; 17(1): 284-300, 2020 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-31794223

RESUMO

Therapeutic protein depots have limited clinical success because of the presence of critical preparation barriers such as low encapsulation, uncontrolled release, and activity loss during processing and storage. In the present study, we used our novel protein-nanoencapsulation (into sugar-glass nanoparticle; SGnP) platform to prepare a protein depot to overcome the abovementioned formidable challenges. The SGnP-mediated microparticle protein depot has been validated using four model proteins (bovine serum albumin, horseradish peroxidase, fibroblastic growth factor, and epidermal growth factor) and model biodegradable poly(lactic-co-glycolic acid) polymer system. The results show that our protein-nanoencapsulation-mediated platform provides a new generic platform to prepare a protein depot through the conventional emulsion method of any polymer and single/multiple protein systems. This protein depot has the required pharmaceutical properties such as high encapsulation efficiency, burst-free sustained release, and protein preservation during processing and storage, making it suitable for off-the-shelf use in therapeutic protein delivery and tissue engineering applications.


Assuntos
Portadores de Fármacos/química , Nanopartículas/química , Proteínas/administração & dosagem , Preparações de Ação Retardada/química , Composição de Medicamentos/métodos , Emulsões , Fator de Crescimento Epidérmico/química , Fator de Crescimento Epidérmico/metabolismo , Fatores de Crescimento de Fibroblastos/química , Fatores de Crescimento de Fibroblastos/metabolismo , Vidro/química , Peroxidase do Rábano Silvestre/química , Peroxidase do Rábano Silvestre/metabolismo , Humanos , Células MCF-7 , Microscopia Eletrônica de Varredura , Nanopartículas/ultraestrutura , Tamanho da Partícula , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Proteínas/química , Soroalbumina Bovina/química , Soroalbumina Bovina/metabolismo , Esferoides Celulares/efeitos dos fármacos , Açúcares
12.
Exp Parasitol ; 208: 107802, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31730782

RESUMO

In insects, diet plays an important role in growth and development. Insects can vary their diet composition based on their physiological needs. In this study we tested the influence of diet composition involving varying concentrations of macronutrients and zinc on the immune-tolerance following parasite and pathogen exposure in Spodoptera litura larvae. We also tested the insecticidal potential of Mesorhabditis belari, Enterobacter hormaechei and its secondary metabolites on Spodoptera litura larvae. The results shows macronutrient composition does not directly affect the larval tolerance to nematode infection. However, Zinc supplemented diet improved the immune tolerance. While larvae exposed to bacterial infection performed better on carbohydrate rich diet. Secondary metabolites from bacteria produced an immune response in dose dependent mortality. The study shows that the larvae maintained on different diet composition show varied immune tolerance which is based on the type of infection.


Assuntos
Enterobacter/fisiologia , Controle Biológico de Vetores , Rhabditoidea/fisiologia , Spodoptera/imunologia , Análise de Variância , Animais , Bioensaio , Carboidratos/administração & dosagem , Cromatografia Líquida de Alta Pressão , Dieta , Enterobacter/imunologia , Enterobacter/patogenicidade , Cromatografia Gasosa-Espectrometria de Massas , Tolerância Imunológica , Larva/imunologia , Dose Letal Mediana , Proteínas/administração & dosagem , Rhabditoidea/imunologia , Rhabditoidea/patogenicidade , Espectroscopia de Infravermelho com Transformada de Fourier , Spodoptera/fisiologia , Simbiose , Virulência , Zinco/administração & dosagem
13.
Int J Pharm ; 573: 118722, 2020 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-31705976

RESUMO

Technology such as the use of microfluidics to generate liposomes has been well researched, yet the stabilisation of liposomal formulations is a major challenge to their greater implementation. To the best of our knowledge, this is the first study investigating the use of 96 well plates to freeze-dry ovalbumin (OVA) loaded neutral (DMPC:Chol and DSPC:Chol), anionic (DSPC:Chol:PS) and cationic (DSPC:Chol:DOTAP) liposomes. Through the use of high throughput screening, a freeze drying cycle was optimised; ramp freezing from from 4 °C to -45 °C, followed by primary drying at -30 °C and secondary drying at 30 °C under a vacuum of 0.1 mBar. These parameters maintained liposome physicochemical properties, with the liposomes remaining below 100 nm and were homogenous (polydispersity index of less than 0.2 post rehydration). Minimal leakage of the OVA protein was observed, with almost 100% OVA remaining encapsulated post rehydration of the formulations. Here we have identified a simple method that allows for the rapid screening and freeze-drying of a range of liposomal formulations.


Assuntos
Sistemas de Liberação de Medicamentos , Microfluídica , Ovalbumina/administração & dosagem , Proteínas/administração & dosagem , Colesterol/química , Dimiristoilfosfatidilcolina/química , Ácidos Graxos Monoinsaturados/química , Liofilização , Ensaios de Triagem em Larga Escala , Lipossomos , Ovalbumina/química , Fosfatidilcolinas/química , Proteínas/química , Compostos de Amônio Quaternário/química , Tecnologia Farmacêutica
14.
Biomater Sci ; 8(4): 1020-1044, 2020 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-31621709

RESUMO

Oral drug delivery remains the most preferred approach due to its multiple advantages. Recently there has been increasing interest in the development of advanced vehicles for oral delivery of different therapeutics. Among them, biomimetic and bioinspired strategies are emerging as novel approaches that are promising for addressing biological barriers encountered by traditional drug delivery systems. Herein we provide a state-of-the-art review on the current progress of biomimetic particulate oral delivery systems. Different biomimetic nanoparticles used for oral drug delivery are first discussed, mainly including ligand/antibody-functionalized nanoparticles, transporter-mediated nanoplatforms, and nanoscale extracellular vesicles. Then we describe bacteria-derived biomimetic systems, with respect to oral delivery of therapeutic proteins or antigens. Subsequently, yeast-derived oral delivery systems, based on either chemical engineering or bioengineering approaches are discussed, with emphasis on the treatment of inflammatory diseases and cancer as well as oral vaccination. Finally, bioengineered plant cells are introduced for oral delivery of biological agents. A future perspective is also provided to highlight the existing challenges and possible resolution toward clinical translation of currently developed biomimetic oral therapies.


Assuntos
Antígenos/administração & dosagem , Biomimética/métodos , Proteínas/administração & dosagem , Administração Oral , Antígenos/química , Bioengenharia , Sistemas de Liberação de Medicamentos , Humanos , Nanopartículas/química , Proteínas/química
15.
Zygote ; 28(1): 32-36, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31601279

RESUMO

The present study evaluated the effect of knockout serum replacement (KSR), fetal bovine serum (FBS) and bovine serum albumin (BSA) on the viability and growth of bovine secondary follicles cultured in vitro for 12 days. To this end, secondary follicles were isolated (185-202 µm) and cultured in vitro in TCM-199+ medium supplemented with KSR (5% and 10%), FBS (5% and 10%) or BSA (3 mg/ml) at 38.5°C with 5% CO2 in air. Follicular diameters were evaluated on days 0, 4, 8 and 12. After 12 days of culture, follicular survival analysis was performing by using calcein-AM and ethidium homodimer. Before and after culture, follicles were fixed in paraformaldehyde for histological evaluation. Follicular diameter at different days of culture were compared using the Kruskal-Wallis test, while the percentages of viable follicles were analyzed by chi-squared test (P < 0.05). Results showed that follicles cultured in the presence of KSR at both concentrations presented higher follicular survival rates than those cultured in control medium alone or supplemented with FBS or BSA. Conversely, the presence of KSR, BSA or FBS did not increase follicular diameter after 12 days of culture. Histology analysis showed that, among the tested treatments, follicles cultured in the presence of KSR had preserved rounded oocytes, juxtaposed granulosa cells and intact basal membrane. In conclusion, supplementation of culture medium with KSR increases the follicular survival of bovine secondary follicles cultured in vitro.


Assuntos
Meios de Cultura/farmacologia , Técnicas de Maturação in Vitro de Oócitos , Oócitos/citologia , Folículo Ovariano/citologia , Proteínas/administração & dosagem , Soroalbumina Bovina/administração & dosagem , Animais , Bovinos , Feminino , Células da Granulosa/citologia , Células da Granulosa/efeitos dos fármacos , Células da Granulosa/metabolismo , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/metabolismo
16.
Aging Clin Exp Res ; 32(4): 605-616, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31183750

RESUMO

BACKGROUND: Adaptation to strength training in very old mobility-limited individuals is not fully characterized. Therefore, the aim of this study was to perform a thorough investigation of the adaptation to a lower body strength training regime in this population, with particular emphasis on the relationship between changes in selected variables. METHODS: Twenty-two mobility-limited older men and women (85 ± 6 years) were randomized to either a group performing 30 min of heavy-load strength training three times a week, with daily protein supplementation, for 10 weeks (ST), or a control group. End points were leg lean mass assessed by DXA, muscle thickness assessed by ultrasound, isometric and dynamic strength, rate of torque development, and functional capacity. RESULTS: Leg lean mass increased from baseline in ST (0.7 ± 0.3 kg), along with increased thickness of vastus lateralis (4.4 ± 3.2%), rectus femoris (6.7 ± 5.1%), and vastus intermedius (5.8 ± 5.9%). The hypertrophy was accompanied by improved knee extensor strength (20-23%) and functional performance (7-11%). In ST, neither the change in leg lean mass nor muscle thickness correlated with changes in muscle strength. However, a strong correlation was observed between the change in isometric strength and gait velocity (r = 0.70). CONCLUSIONS: The mismatch between gains in muscle size and strength suggests that muscle quality-related adaptations contributed to the increases in strength. The correlations observed between improvements in strength and function suggests that interventions eliciting large improvements in strength may also be superior in terms of functional gains in this population.


Assuntos
Suplementos Nutricionais , Força Muscular/fisiologia , Desempenho Físico Funcional , Proteínas/administração & dosagem , Treinamento de Resistência/métodos , Idoso , Idoso de 80 Anos ou mais , Feminino , Fragilidade/prevenção & controle , Humanos , Masculino , Limitação da Mobilidade
17.
Eur J Pharm Biopharm ; 147: 10-18, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31837391

RESUMO

Low volume aseptic filling of parenterals, particularly monoclonal antibodies is becoming increasingly important with the development of more and more intravitreal drugs and high concentrated formulations. Especially monoclonal antibodies are very delicate products to fill and the use of the right fill finish equipment plays an important role during process development. Protein aggregation can occur under conditions described in literature and can be influenced by the fill finish processing. The mechanism of product stress inside the filling systems is yet not fully understood. This study evaluated three different dosing systems to assess protein degradation caused by the shear rate during low volume filling of monoclonal antibodies. The newly developed quantitative liposomal shear stress model revealed the highest shear rate in the radial peristaltic pump, followed by the rotary piston pump and the linear peristaltic pump. In contrast to that, we found the highest sub-visible particle counts (>2 µm) in the rotary piston pump. We used computational fluid dynamics for a better and deeper understanding of filling processes inside the different dosing systems. Our results document that the rotary piston pump creates a recirculation zone inside the cylinder, where the protein formulation could be trapped and be exposed to the shear stress multiple times resulting in a cumulative shearing. This finding could serve as an explanation for the highest sub-particle counts in low volume filling using a rotary piston pump.


Assuntos
Anticorpos Monoclonais/química , Composição de Medicamentos/métodos , Proteínas/química , Tecnologia Farmacêutica/métodos , Anticorpos Monoclonais/administração & dosagem , Composição de Medicamentos/instrumentação , Desenho de Equipamento , Hidrodinâmica , Lipossomos , Proteínas/administração & dosagem , Proteínas/normas , Esterilização , Estresse Mecânico , Tecnologia Farmacêutica/instrumentação
18.
Drug Metab Dispos ; 47(12): 1443-1456, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31748266

RESUMO

For therapeutic proteins, the currently established standard development path generally does not foresee biotransformation studies by default because it is well known that the clearance of therapeutic proteins proceeds via degradation to small peptides and individual amino acids. In contrast to small molecules, there is no general need to identify enzymes involved in biotransformation because this information is not relevant for drug-drug interaction assessment and for understanding the clearance of a therapeutic protein. Nevertheless, there are good reasons to embark on biotransformation studies, especially for complex therapeutic proteins. Typical triggers are unexpected rapid clearance, species differences in clearance not following the typical allometric relationship, a mismatch in the pharmacokinetics/pharmacodynamics (PK/PD) relationship, and the need to understand observed differences between the results of multiple bioanalytical methods (e.g., total vs. target-binding competent antibody concentrations). Early on during compound optimization, knowledge on protein biotransformation may help to design more stable drug candidates with favorable in vivo PK properties. Understanding the biotransformation of a therapeutic protein may also support designing and understanding the bioanalytical assay and ultimately the PK/PD assessment. Especially in cases where biotransformation products are pharmacologically active, quantification and assessment of their contribution to the overall pharmacological effect can be important for establishing a PK/PD relationship and extrapolation to humans. With the increasing number of complex therapeutic protein formats, the need for understanding the biotransformation of therapeutic proteins becomes more urgent. This article provides an overview on biotransformation processes, proteases involved, strategic considerations, regulatory guidelines, literature examples for in vitro and in vivo biotransformation, and technical approaches to study protein biotransformation. SIGNIFICANCE STATEMENT: Understanding the biotransformation of complex therapeutic proteins can be crucial for establishing a pharmacokinetic/pharmacodynamic relationship. This article will highlight scientific, strategic, regulatory, and technological features of protein biotransformation.


Assuntos
Preparações Farmacêuticas/metabolismo , Proteínas/farmacocinética , Bibliotecas de Moléculas Pequenas/farmacocinética , Animais , Biotransformação , Interações Medicamentosas , Humanos , Preparações Farmacêuticas/administração & dosagem , Proteínas/administração & dosagem , Proteínas/farmacologia , Bibliotecas de Moléculas Pequenas/administração & dosagem , Bibliotecas de Moléculas Pequenas/farmacologia
19.
Cells ; 8(12)2019 11 25.
Artigo em Inglês | MEDLINE | ID: mdl-31775322

RESUMO

Extracellular vesicles (EVs) are mediators of intercellular communication by transferring functional biomolecules from their originating cells to recipient cells. This intrinsic ability has gained EVs increased scientific interest in their use as a direct therapeutic in the field of regenerative medicine or as vehicles for drug delivery. EVs derived from stem cells or progenitor cells can act as paracrine mediators to promote repair and regeneration of damaged tissues. Despite substantial research efforts into EVs for various applications, their use remains limited by the lack of highly efficient and scalable production methods. Here, we present the biofabrication of cell-derived nanovesicles (NVs) as a scalable, efficient, and cost-effective production alternative to EVs. We demonstrate that NVs have a comparable size and morphology as EVs, but lack standard EV (surface) markers. Additionally, in vitro uptake experiments show that human fetal cardiac fibroblast, endothelial cells, and cardiomyocyte progenitor cells internalize NVs. We observed that cardiac progenitor cell-derived NVs and EVs are capable of activating mitogen-activated protein kinase 1/2 (MAPK1/2)-extracellular signal-regulated kinase, and that both NVs and EVs derived from A431 and HEK293 cells can functionally deliver Cre-recombinase mRNA or protein to other cells. These observations indicate that NVs may have similar functional properties as EVs. Therefore, NVs have the potential to be applied for therapeutic delivery and regenerative medicine purposes.


Assuntos
Portadores de Fármacos/farmacologia , Células Endoteliais/efeitos dos fármacos , Miócitos Cardíacos/efeitos dos fármacos , Nanopartículas/uso terapêutico , Células-Tronco/citologia , Células Endoteliais/citologia , Células HEK293 , Humanos , Miócitos Cardíacos/citologia , Proteínas/administração & dosagem , RNA Mensageiro/administração & dosagem , Medicina Regenerativa/métodos
20.
Int J Pharm ; 572: 118720, 2019 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-31715357

RESUMO

Therapeutic proteins are labile macromolecules that are prone to degradation during production, freeze-drying and storage. Recent studies showed that nanoparticles can enhance the stability and oral bioavailability of encapsulated proteins. Several conventional approaches (enzyme inhibitors, mucoadhesive polymers) and novel strategies (surface modification, ligand conjugation, flash nano-complexation, stimuli-responsive drug delivery systems) have been employed to improve the physiochemical properties of nanoparticles such as size, zeta potential, morphology, polydispersity index, drug release kinetics and cell-targeting capacity. However, clinical translation of protein-based nanoparticle is limited due to poor experimental design, protocol non-compliance and instrumentation set-up that do not reflect the physiological conditions, resulting in difficulties in mass production of nanoparticles and waste in research funding. In order to address the above concerns, we conducted a comprehensive review to examine the experimental designs and conditions for physical characterization of protein-based nanoparticles. Reliable and robust characterization is essential to verify the cellular interactions and therapeutic potential of protein-based nanoparticles. Importantly, there are a number of crucial factors, which include sample treatment, analytical method, dispersants, sampling grid, staining, quantification parameters, temperature, drug concentration and research materials, should be taken into careful consideration. Variations in research protocol and unreasonable conditions that are used in optimization of pharmaceutical formulations can have great impact in result interpretation. Last but not least, we reviewed all novel instrumentations and assays that are available to examine mucus diffusion capacity, stability and bioactivity of protein-based nanoparticles. These include circular dichroism, fourier transform infrared spectroscopy, X-ray diffractogram, UV spectroscopy, differential scanning calorimetry, fluorescence spectrum, Förster resonance energy transfer, NMR spectroscopy, Raman spectroscopy, cellular assays and animal models.


Assuntos
Nanopartículas/química , Proteínas/química , Administração Oral , Animais , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Humanos , Mucinas/química , Nanopartículas/administração & dosagem , Nanotecnologia , Tamanho da Partícula , Proteínas/administração & dosagem , Projetos de Pesquisa
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