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1.
J Chem Inf Model ; 61(1): 525-534, 2021 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-33426873

RESUMO

Blood-brain barrier peptides (BBPs) have a large range of biomedical applications since they can cross the blood-brain barrier based on different mechanisms. As experimental methods for the identification of BBPs are laborious and expensive, computational approaches are necessary to be developed for predicting BBPs. In this work, we describe a computational method, BBPpred (blood-brain barrier peptides prediction), that can efficiently identify BBPs using logistic regression. We investigate a wide variety of features from amino acid sequence information, and then a feature learning method is adopted to represent the informative features. To improve the prediction performance, seven informative features are selected for classification by eliminating redundant and irrelevant features. In addition, we specifically create two benchmark data sets (training and independent test), which contain a total of 119 BBPs from public databases and the literature. On the training data set, BBPpred shows promising performances with an AUC score of 0.8764 and an AUPR score of 0.8757 using the 10-fold cross-validation. We also test our new method on the independent test data set and obtain a favorable performance. We envision that BBPpred will be a useful tool for identifying, annotating, and characterizing BBPs. BBPpred is freely available at http://BBPpred.xialab.info.

2.
Environ Pollut ; : 116106, 2020 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-33272795

RESUMO

This study performs an environmental risk assessment (ERA) of the anthelmintic medicine albendazole (ABZ) in the eastern African region. A systematic literature search strategy was applied to obtain quantitative information on the physicochemical characteristics, the metabolization-fate, the ecotoxicity and the environmental occurrence in different countries worldwide serving as model regions. In addition, insilico tools were employed to obtain data on physicochemical characteristics and toxic hazards of ABZ and its metabolites. Moreover, ERA models were used to predict environmental concentrations in different compartments and compare them with the measured environmental concentrations. Finally, the environmental risk of ABZ in the eastern Africa was estimated by calculating the risk quotient (RQ), and its uncertainty estimated by Monte Carlo simulation. The predicted environmental concentrations of ABZ in surface water in the model region based on consumption (1.6-267 ng/L) were within the range of values obtained from the measured environmental concentrations of the same region (0.05-101,000 ng/L). Using these models with adapted input variables for eastern Africa, the predicted surface water concentration in that region was 19,600 ± 150 ng/L (95% CI). The calculated soil concentrations of ABZ in the model regions and the eastern Africa were found to be 0.057 ± 0.0 µg/kg and 0.022 ± 0.0 µg/kg, respectively. The environmental risk expressed as risk quotient of ABZ in eastern Africa estimated for the aquatic compartment (146 ± 1) indicated a significant environmental risk calling on appropriate actions from the competent authorities to reduce this risk in this region.

3.
Front Pharmacol ; 11: 1336, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32982746

RESUMO

Upregulation of the RAS-RAF-MEK-ERK-MAPK pathway is involved in the development of several human tumors, aortic aneurysms, atherosclerosis, and cardiomyopathy. Refametinib, a highly selective MEK-inhibitor, has already shown antineoplastic activity in phase II trials. Furthermore, it showed potency to attenuate aortic root growth in murine models. Current formulations of this drug however necessitate oral gavage as a delivery method for long-term studies, which is labor-intensive and induces stress and occasional injury, potentially confounding results. Therefore, we developed a novel oral administration method for refametinib. A 2-hydroxypropyl-beta-cyclodextrin (HPBCD) based drinking water preparation of refametinib was formulated, for which a selective, analytical UHPLC-UV method was developed to assess the in-use stability. Next, 16 week old male wild-type C57Bl/6J mice received either a daily dose of 50 or 75 mg/kg/day refametinib or were given regular drinking water during 7 days. In both dosage groups the refametinib plasma levels were measured (n = 10 or 7, respectively). Furthermore, pERK/total ERK protein levels were calculated in the myocardial and aortic tissue of mice receiving a daily dose of 50 mg/kg/day refametinib and untreated mice (n = 4/group). After 7 days no significant degradation of refametinib was observed when dissolved in drinking water provided that drinking bottles were protected from UV/visible light. Furthermore, a dose-dependent increase in refametinib plasma levels was found whereby active plasma levels (> 1.2 µg/mL) were obtained even in the lowest dose-group of 50 mg/kg/day. A significant reduction of pERK/total ERK protein levels compared to untreated mice was observed in aortic and myocardial tissue of mice receiving a daily dose of 50 mg/kg/day refametinib. Importantly, a relatively high mortality rate was noted in the highest dose group (n = 5). This approach provides a valid alternative oral administration method for refametinib with a reduced risk of complications due to animal manipulation and without loss of functionality, which can be implemented in future research regarding the malignant upregulation of the RAS-RAF-MEK-ERK-MAPK pathway. However, care must be taken not to exceed the toxic dose.

4.
J Biol Inorg Chem ; 25(6): 875-885, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32719971

RESUMO

L-ascorbic acid 2-phosphate magnesium (APMg) salt is a vitamin C derivative frequently used as a raw material in cell and tissue therapy. APMg is not only used as a replacement of the unstable ascorbate, but also shows additional cell-biological functionalities. However, its unknown structural characteristics hamper the mechanistic elucidation of its biological role. Therefore, different techniques were applied for APMg structure characterization. Firstly, the stoichiometric composition was characterized by its solvent, ligand and magnesium content. No crystals of APMg could be obtained; however, a single crystal of APNa, the sodium salt of l-ascorbic acid 2-phosphate, was successfully obtained and its crystal structure was elucidated. FT-IR was applied to further clarify the structure of solid APMg. Finally, the structure of APMg in aqueous solution was explored by potentiometric titration as well as FT-IR.

5.
ACS Omega ; 5(26): 16120-16127, 2020 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-32656434

RESUMO

Finding adequate biomarkers for rapid and accurate disease detection, prognosis, and therapy is increasingly important. Quorum-sensing peptides are herein a new emerging group, produced by bacteria, fungi, protozoa, and viruses, with blood being the most straightforward sample type to detect/quantitate them. However, detailed information about suitable blood sample collection methods and storage conditions for measuring these quorum-sensing peptides hampers further clinical research and development. Here, we first tested the time-dependent stability of a set of chemically diverse quorum-sensing peptides, spiked in blood at different temperatures (4, 21, and 37 °C) in four different ethylenediamine tetraacetic acid (EDTA)-containing plasma tubes (with different protein-stabilizing additives) over a period of up to 7.5 h. Next, we determined the storage stability of these quorum-sensing peptides in plasma at different temperatures (4, -35, and -80 °C). UPLC/MS-MS was used to selectively detect and quantify the spiked quorum-sensing peptides. The results of this study indicate that a cost-effective tube, designed for traditional proteomics and stored at 4 °C, is the preferred collection condition when quorum-sensing peptides need to be detected/quantified in human plasma. When the tubes are handled at room temperature (21 °C), a more specialized tube is required. Long-term storage of plasma samples, even under low-temperature conditions (-80 °C), indicates rapid degradation of certain quorum-sensing peptides.

6.
J Am Med Dir Assoc ; 21(7): 909-914.e2, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32674818

RESUMO

OBJECTIVES: Angiotensin-converting enzyme inhibitors (ACEi), angiotensin II receptor blockers (ARBs), and HMG-CoA reductase inhibitors ("statins") have been hypothesized to affect COVID-19 severity. However, up to now, no studies investigating this association have been conducted in the most vulnerable and affected population groups (ie, older adults residing in nursing homes). The objective of this study was to explore the association of ACEi/ARB and/or statins with clinical manifestations in COVID-19-infected older adults residing in nursing homes. DESIGN: We undertook a retrospective multicenter cohort study to analyze the association between ACEi/ARB and/or statin use with clinical outcome of COVID-19. The outcomes were (1) serious COVID-19 defined as long-stay hospital admission or death within 14 days of disease onset, and (2) asymptomatic (ie, no disease symptoms in the whole study period while still being diagnosed by polymerase chain reaction). SETTING AND PARTICIPANTS: A total of 154 COVID-19-positive subjects were identified, residing in 1 of 2 Belgian nursing homes that experienced similar COVID-19 outbreaks. MEASURES: Logistic regression models were applied with age, sex, functional status, diabetes, and hypertension as covariates. RESULTS: We found a statistically significant association between statin intake and the absence of symptoms during COVID-19 (odds ratio [OR] 2.91; confidence interval [CI] 1.27-6.71), which remained statistically significant after adjusting for covariates (OR 2.65; CI 1.13-6.68). Although the effects of statin intake on serious clinical outcome were in the same beneficial direction, these were not statistically significant (OR 0.75; CI 0.24-1.87). There was also no statistically significant association between ACEi/ARB and asymptomatic status (OR 2.72; CI 0.59-25.1) or serious clinical outcome (OR 0.48; CI 0.10-1.97). CONCLUSIONS AND IMPLICATIONS: Our data indicate that statin intake in older, frail adults could be associated with a considerable beneficial effect on COVID-19 clinical symptoms. The role of statins and renin-angiotensin system drugs needs to be further explored in larger observational studies as well as randomized clinical trials.


Assuntos
Antagonistas de Receptores de Angiotensina/administração & dosagem , Inibidores da Enzima Conversora de Angiotensina/administração & dosagem , Infecções por Coronavirus/tratamento farmacológico , Infecções por Coronavirus/epidemiologia , Inibidores de Hidroximetilglutaril-CoA Redutases/administração & dosagem , Pneumonia Viral/tratamento farmacológico , Pneumonia Viral/epidemiologia , Idoso , Idoso de 80 Anos ou mais , Bélgica/epidemiologia , Causas de Morte , Estudos de Coortes , Feminino , Avaliação Geriátrica , Instituição de Longa Permanência para Idosos/estatística & dados numéricos , Humanos , Modelos Logísticos , Masculino , Casas de Saúde/estatística & dados numéricos , Razão de Chances , Pandemias/estatística & dados numéricos , Estudos Retrospectivos , Medição de Risco , Índice de Gravidade de Doença , Taxa de Sobrevida , Resultado do Tratamento
7.
Malar J ; 19(1): 139, 2020 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-32264882

RESUMO

BACKGROUND: Dissolution of artemether (ART) and lumefantrine (LUM) active pharmaceutical ingredients (APIs) in fixed dose combination (FDC) ART/LUM tablets is one of the critical quality attributes. Thus, the verification of the release profile of ART and LUM from FDC ART/LUM tablets using a robust and discriminatory dissolution method is crucial. Therefore, the aim of this study was to develop and validate an appropriate dissolution method for quality control of FDC ART/LUM tablets. METHODS: The dissolution medium was selected based on saturation solubility data and sink conditions. The effect of agitation speed, pH and surfactant concentration on the release of ART and LUM was evaluated by employing a two-level factorial experiment. The resulting final method was validated for linearity, precision, robustness and API stability. In addition, the discriminatory power of the method was evaluated using expired and unexpired FDC ART/LUM products. RESULTS: A suitable dissolution profile of FDC ART/LUM tablets was obtained in 900 ml HCl (0.025 N, pH 1.6) with 1%Myrj 52 using paddle method at 100 rpm and 37 °C. ART and LUM were analysed using a HPLC method with UV detection at wavelengths of 210 and 335 nm, respectively. The results from the stability study showed that ART and LUM were sufficiently stable in HCl (0.025 N, pH 1.6) with 1%Myrj 52 at 37 °C. The method was linear (r2 = 0.999) over the concentration range of 6.25-100 µg/ml. The results for precision were within the acceptance limit (%RSD < 2). The percent relative standard deviation (< 2%) and statistically non-significant (p > 0.05) difference in release of ART and LUM observed between deliberately changed dissolution method settings (pH = 1.6 ± 0.2 or agitation speed = 100 ± 2) and optimized dissolution conditions revealed the robustness of the dissolution method. The method was capable to discriminate among different FDC ART/LUM products with different quality. CONCLUSIONS: The developed dissolution method is robust and discriminatory. It can be used in the quality evaluation of FDC ART/LUM tablets.


Assuntos
Antimaláricos/química , Combinação Arteméter e Lumefantrina/química , Liberação Controlada de Fármacos , Antimaláricos/análise , Combinação Arteméter e Lumefantrina/análise , Controle de Qualidade , Solubilidade , Solventes , Comprimidos
8.
Drug Test Anal ; 12(1): 67-77, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31471998

RESUMO

Thermal sensitivity, as a practical measure of thermostability, is an interesting quality attribute that can be used in the quality control (QC) release of biopharmaceuticals. This article investigates circular dichroism (CD) spectroscopy and nano-differential scanning fluorimetry (nano-DSF) to evaluate the thermal stability of E.coli L-asparaginase (L-ASNase) for QC purposes. In CD, molar ellipticity as a function of temperature (from 20 to 80°C) was measured at 222 nm. Different L-ASNase samples dissolved in different diluents were investigated by determining the melting temperature (Tm ) from the first derivative curve as well as the slope of the fitted sigmoidal function of the temperature gradient CD data. The obtained Tm values could be correlated with the L-ASNase sample origin as well as with the pH of the diluent. The Tm values obtained from the CD data were moreover consistent with the Tm values determined by nano-DSF, confirming their reliability. Next to the Tm value, also the slope of the fitted sigmoidal CD-function was able to differentiate different L-ASNase samples, including unstressed from stressed protein. By using both the Tm and the curve slope, the thermal stability of L-ASNase was investigated, demonstrating and recommending the use of this heat-stress characteristic as a QC quality attribute of proteins, which can be applied to detect substandard and falsified proteins.


Assuntos
Antineoplásicos/química , Asparaginase/química , Escherichia coli/enzimologia , Dicroísmo Circular , Estabilidade de Medicamentos , Estabilidade Enzimática , Escherichia coli/química , Fluorometria , Concentração de Íons de Hidrogênio , Cinética , Controle de Qualidade , Temperatura
9.
Biochim Biophys Acta Mol Basis Dis ; 1866(3): 165646, 2020 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-31870715

RESUMO

Skeletal muscle makes up the largest part of human body mass and a good maintenance of this organ is essential for general health. In accordance, muscle wasting, a frequent phenomenon in many diseases, is associated with functional decline and a decrease in quality of life. Unfortunately, due to a lack of knowledge of the underlying pathophysiology, no targeted therapies exist today to encounter muscle wasting. Recent studies suggest a role for the gut microbiome in muscle wasting, without the mediators of this gut-muscle axis being identified. Here we evaluated the possible effects of 75 quorum sensing molecules (QSM), traditionally only seen as intra-bacterial communication molecules, on C2C12 muscle cells, studying viability, differentiation, inflammation, mitochondrial changes and protein degradation as biological outcomes. The responses were evaluated using different approaches: median absolute deviation, quartiles, strictly standardized mean difference and robust strictly standardized mean difference. This study resulted in 30 QSM, with effects observed on C2C12 cells. Known producers of the 27 peptide QSM belong to species of the genus Staphylococcus, Streptococcus, Enterococcus, Bacillus, Lactobacillus and Escherichia, while the 3 non-peptide QSM are produced by a broad range of Gram-positive and Gram-negative bacteria. Altogether, these proof-of-concept findings provide the first evidence that QSM produced by microbiota play a role in the gut-muscle axis, opening new perspectives for diagnostic and therapeutic targets in muscle wasting diseases.


Assuntos
Bactérias/metabolismo , Microbiota/fisiologia , Células Musculares/metabolismo , Percepção de Quorum/fisiologia , Animais , Diferenciação Celular/fisiologia , Linhagem Celular , Sobrevivência Celular/fisiologia , Microbioma Gastrointestinal/fisiologia , Inflamação/metabolismo , Camundongos , Mitocôndrias/metabolismo , Músculo Esquelético/metabolismo , Qualidade de Vida
10.
Anal Chem ; 92(2): 1712-1719, 2020 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-31874035

RESUMO

Analytical method development for peptides often proves challenging since these molecules can adsorb to the plastic or glass consumables used in the analysis. This adsorption causes considerable loss and unreliable results, especially in the lower concentration range. Therefore, a variety of antiadsorption strategies have previously been developed to cope with this adsorption, often however incompatible with direct liquid chromatography-mass spectrometry (LC-MS) analysis. Here, a novel antiadsorption diluent is introduced, based on controlled hydrolysis and precipitation of bovine serum albumin. This diluent considerably decreases the adsorption of certain peptides to glass. Moreover, it is LC-MS compatible and can also be used in combination with formic acid and/or acetonitrile addition.

11.
Q J Nucl Med Mol Imaging ; 64(1): 105-114, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29697217

RESUMO

BACKGROUND: Chemical modifications such as PEG, polyamine and radiolabeling on proteins can alter their pharmacokinetic behavior and their blood-brain barrier (BBB) transport characteristics. NOTA, i.e. 1,4,7-triazacyclononane-1,4,7-triacetic acid, is a bifunctional chelating agent that has attracted the interest of the scientific community for its high complexation constant with metals like gallium. Until now, the comparative BBB transport characteristics of NOTA-modified proteins versus unmodified proteins are not yet described. METHODS: Somatropin (i.e. recombinant human growth hormone), NOTA-conjugated somatropin and gallium-labelled NOTA-conjugated somatropin were investigated for their brain penetration characteristics (multiple time regression and capillary depletion [CD]) in an in vivo mice model to determine the blood-brain transfer properties. RESULTS: The three compounds showed comparable initial brain influx, with Kin=0.38±0.14 µL/(g×min), 0.36±0.16 µL/(g×min) and 0.28±0.18 µL/(g×min), respectively. CD indicated that more than 80% of the influxed compounds reached the brain parenchyma. All three compounds were in vivo stable in serum and brain during the time frame of the experiments. CONCLUSIONS: Our results show that modification of NOTA as well as gallium chelation onto proteins, in casu somatropin, does not lead to a significantly changed pharmacokinetic profile at the blood-brain barrier.

12.
Biochim Biophys Acta Mol Basis Dis ; : 165585, 2019 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-31678164

RESUMO

This article has been withdrawn at the request of the author for administrative reasons. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at https://www.elsevier.com/about/our-business/policies/article-withdrawal.

13.
Malar J ; 18(1): 236, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31307475

RESUMO

BACKGROUND: Malaria caused by Plasmodium vivax and Plasmodium falciparum is among the major public health problems in most endemic areas of the world. Artemisinin-based combination therapy (ACT) has been recommended as a first-line treatment for uncomplicated Plasmodium falciparum malaria almost in all endemic regions. Since ineffectively regulated medicines in resource limited settings could favour infiltration of poor quality anti-malarial medicines into pharmaceutical supply chain and jeopardize a positive treatment outcome, regular monitoring of the quality of anti-malarial medicines is critical. Thus, the aim of this study was to assess the quality of fixed dose combination (FDC) artemether (ART)/lumefantrine (LUM) tablets available in Jimma zone, Ethiopia. METHODS: This study was conducted in Jimma zone, Ethiopia. A total of 74 samples of FDC ART/LUM (20 mg ART/120 mg LUM) tablets were collected from 27 public facilities. All samples were subjected to visual inspection and the relevant information was recorded. The samples were transported to Jimma University Laboratory of Drug Quality (JuLaDQ) and stored at ambient temperature (20 °C to 25 °C) until analysis. The Pharmacopoeial conform/non-conform methods and the risk-based Derringer's desirability function approach were employed to assess the pharmaceutical quality of the investigated products. RESULTS: The visual inspection results revealed that there were no signs of falsified in the investigated products. Identification test results of samples indicated that all samples contained the stated active pharmaceutical ingredients (APIs). The results of uniformity of mass indicated that all samples complied with International Pharmacopoeial specification limits. The assay results, expressed as percent label claim (%lc) of ART (89.8 to 108.8%, mean ± SD = 99.1 ± 3.9%) and LUM (90.0 to 111.9%, mean ± SD = 98.2 ± 3.8%) revealed that, all samples complied with International Pharmacopoeia acceptance specification limits (i.e. 90-110%lc), except one generic product (IPCA Laboratories Ltd., India) which contains excessive LUM (111.9 ± 1.7%lc). The risk priority number (RPN) results revealed that assay (RPN = 392) is relatively the most critical quality attribute followed by identity (RPN = 280) and mass uniformity (40). Quality evaluation based on psycho-physical Harrington's scale revealed that more than 96% of samples were within the acceptable ranges (D ≥ 0.7-1.0). CONCLUSIONS: Both Pharmacopoeial and risk-based desirability function approaches to quality evaluation applied to the investigated products revealed that above 96% FDC ART/LUM tablets circulating in public settings of Jimma zone are of good quality.


Assuntos
Antimaláricos/análise , Combinação Arteméter e Lumefantrina/análise , Malária Falciparum/tratamento farmacológico , Etiópia
14.
Clin Epigenetics ; 11(1): 101, 2019 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-31300053

RESUMO

Peptides originating from different sources (endogenous, food derived, environmental, and synthetic) are able to influence different aspects of epigenetic regulation. Endogenous short peptides, resulting from proteolytic cleavage of proteins or upon translation of non-annotated out of frame transcripts, can block DNA methylation and hereby regulate gene expression. Peptides entering the body by digestion of food-related proteins can modulate DNA methylation and/or histone acetylation while environmental peptides, synthesized by bacteria, fungi, and marine sponges, mainly inhibit histone deacetylation. In addition, synthetic peptides that reverse or inhibit different epigenetic modifications of both histones and the DNA can be developed as well. Next to these DNA and histone modifications, peptides can also influence the expression of non-coding RNAs such as lncRNAs and the maturation of miRNAs.Seen the advantages over small molecules, the development of peptide therapeutics is an interesting approach to treat diseases with a strong epigenetic basis like cancer and Alzheimer's disease. To date, only a limited number of drugs with a proven epigenetic mechanism of action have been approved by the FDA of which two (romidepsin and nesiritide) are peptides. A large knowledge gap concerning epigenetic effects of peptides is present, and this class of molecules deserves more attention in the development as epigenetic modulators. In addition, none of the currently approved peptide drugs are under investigation for their potential effects on epigenetics, hampering drug repositioning of these peptides to other indications with an epigenetic etiology.


Assuntos
Epigênese Genética/efeitos dos fármacos , Peptídeos/farmacologia , Acetilação/efeitos dos fármacos , Metilação de DNA/efeitos dos fármacos , Histonas/metabolismo , Humanos
15.
Talanta ; 203: 9-15, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31202354

RESUMO

Biosensors are rising technologies in the pharmaceutical field for medicine discovery, development and Quality Control (QC) stages. Surface acoustic wave (SAW) biosensor employs acoustic waves generated by oscillating a piezoelectric crystal quartz plate to meas. mass and viscosity, and allows to detect and quantify binding events between the analyte and an immobilized interacting ligand. We present here a SAW biosensor based approach for the functional quantification of Escherichia colil-asparaginase (E. colil-ASNase), using polyclonal antibody (pAb) as the interaction partner immobilized on the chip. Different immobilization strategies of pAb were initially evaluated, resulting in the BS3 activated amide coupling via protein G strategy as the final immobilization method. The method was validated by evaluating the selectivity, linearity, as well as accuracy (a recovery of 102.4%) and precision (RSD of 8.5%). The application of the validated method on different samples encompassing different lots of E. colil-ASNase, deamidated E. colil-ASNase and dry-heated E. colil-ASNase (80 °C, 10 min) indicated the suitability of the developed SAW method to quantify E. colil-ASNase. We suggest this SAW method can be adopted as a pharmaceutical QC method.


Assuntos
Asparaginase/análise , Escherichia coli/enzimologia , Animais , Anticorpos Imobilizados/imunologia , Asparaginase/imunologia , Técnicas Biossensoriais/métodos , Ensaios Enzimáticos/métodos , Ouro/química , Limite de Detecção , Coelhos , Som
16.
Protein Pept Lett ; 26(9): 691-701, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31215364

RESUMO

BACKGROUND: Both biodegradable and non-biodegradable peptide-loaded implants are already developed for the long-term treatment of patients, thereby reducing the frequency of drug administration. To further improve peptide formulation, extending the scope of implant-based drug delivery systems towards other polymers and processing techniques is highly interesting. OBJECTIVE: In this study, as a proof-of-principle, the feasibility of hot-melt processing of a peptide active pharmaceutical ingredient was assessed by developing a non-biodegradable poly(ethylenevinyl acetate) (33% VA) implant loaded with 20% (w/w) buserelin acetate. METHODS: Cross-sectional implant characterization was performed by Raman microscopy. The stability of buserelin acetate in the polymeric matrix was evaluated for 3 months under ICH stability conditions and the quantity as well as the degradation products analyzed using LC-UV methods. An in vitro dissolution study was performed as well and buserelin acetate and its degradants analyzed using the same chromatographic methods. RESULTS: No significant quantities of buserelin acetate-related degradation products were formed during the hot-melt preparation as well as during the stability study. Together with the consistent buserelin acetate assay values over time, chemical peptide stability was thus demonstrated. The in vitro buserelin acetate release from the implant was found to be diffusion-controlled after an initial burst release, with stable release profiles in the stability study, demonstrating the functional stability of the peptide implant. CONCLUSION: These results indicate the feasibility of preparing non-biodegradable peptide-loaded implants using the hot-melt production method and may act as a proof of principle concept for further innovation in peptide medicinal formulations.


Assuntos
Busserrelina/química , Portadores de Fármacos/química , Implantes de Medicamento/química , Polivinil/química , Busserrelina/administração & dosagem , Composição de Medicamentos/métodos , Implantes de Medicamento/administração & dosagem , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Estudos de Viabilidade , Temperatura Alta , Estudo de Prova de Conceito , Solubilidade
17.
Talanta ; 201: 259-265, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31122421

RESUMO

Medicines are meant to help people and treat their conditions and to promote general well-being of all members of the society. Unfortunately, this is being compromised by the distribution and sale of poor-quality medicines around the world, being a consequence of non-GMP manufacturing. In this study, the contamination of the outer primary packaging with active pharmaceutical ingredient (API, i.e. artemether) is investigated as a possible and objective, quantifiable marker for GMP-compliance. First, an analytical UPLC-MS method was developed and verified for artemether, with emphasis on the quantification in the lower concentration range. Second, a swabbing procedure for the outer surface of plastic bottles (powders for suspension) was developed, including a swabbing recovery of the API from the bottle surfaces. Finally, twenty antimalarial samples were investigated. All of them showed some degree of outer contamination; however, large differences in the amount of API contamination between the different samples was observed, ranging between 4 and 144 ng/cm2. A positive correlation was found between the amount of artemether on the packaging and the number of information elements missing on the packaging or leaflet, which was used as one of the tools to evaluate the GMP status of the manufacturer.


Assuntos
Antimaláricos/análise , Artemeter/análise , Cromatografia Líquida/métodos , Embalagem de Medicamentos/normas , Espectrometria de Massas/métodos
18.
J Pharm Biomed Anal ; 165: 338-345, 2019 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-30579234

RESUMO

l-ascorbic acid 2-phosphate magnesium (APMg) salt is a vitamin C derivative frequently used in cell culture media for research purposes. It is also used as a raw material in the GMP-manufacturing of gene-, cell- and tissue advanced therapy medicinal products (ATMPs). However, quality methods are currently lacking. Therefore, a LC method was developed, based on hydrophilic interaction (HILIC)-ion exchange (IE) mixed-mode liquid chromatography. The final method consisted of an isocratic system with 15 mM KH2PO4 buffer (pH 2.5 with HCl) acetonitrile (30:70, v/v) mobile phase on a zwitterionic HILIC column, containing an hydrophilic ligand embedded cation-exchange functionality and a surface anion-exchange group. A flow rate of 0.4 mL/min and UV detection at 240 nm was applied. The assay method of APMg was validated, obtaining adequate linearity (R2 = 0.999), precision (RSD of 0.49%) and accuracy (overall recovery of 100.4%). The developed method was successfully applied on five currently marketed products from different suppliers, showing different related substance impurity profiles. Using atomic absorption spectroscopy (AAS), magnesium was found to be bound on the stationary phase, requiring a strong mobile phase to rinse the column. Finally, related impurities were identified using MS/MS and high resolution MS, and found to be ascorbic acid as well as ethyl derivatives, which was further confirmed by NMR.


Assuntos
Ácido Ascórbico/análogos & derivados , Cromatografia Líquida/métodos , Espectrometria de Massas em Tandem/métodos , Ácido Ascórbico/análise , Ácido Ascórbico/química , Ácido Ascórbico/normas , Interações Hidrofóbicas e Hidrofílicas , Controle de Qualidade , Reprodutibilidade dos Testes , Espectrofotometria Atômica/métodos
19.
Nat Mater ; 17(11): 971-977, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30349030

RESUMO

Zika virus is a mosquito-borne virus that is associated with neurodegenerative diseases, including Guillain-Barré syndrome1 and congenital Zika syndrome2. As Zika virus targets the nervous system, there is an urgent need to develop therapeutic strategies that inhibit Zika virus infection in the brain. Here, we have engineered a brain-penetrating peptide that works against Zika virus and other mosquito-borne viruses. We evaluated the therapeutic efficacy of the peptide in a lethal Zika virus mouse model exhibiting systemic and brain infection. Therapeutic treatment protected against mortality and markedly reduced clinical symptoms, viral loads and neuroinflammation, as well as mitigated microgliosis, neurodegeneration and brain damage. In addition to controlling systemic infection, the peptide crossed the blood-brain barrier to reduce viral loads in the brain and protected against Zika-virus-induced blood-brain barrier injury. Our findings demonstrate how engineering strategies can be applied to develop peptide therapeutics and support the potential of a brain-penetrating peptide to treat neurotropic viral infections.


Assuntos
Antivirais/uso terapêutico , Encéfalo/metabolismo , Peptídeos/uso terapêutico , Infecção por Zika virus/tratamento farmacológico , Animais , Antivirais/farmacocinética , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Peptídeos/farmacocinética
20.
Regul Toxicol Pharmacol ; 98: 215-223, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30071240

RESUMO

N-alkylamides (NAAs) are secondary metabolites occurring in more than 25 plant families. Plants containing NAAs are traditionally used in food for flavouring, tingling, pungent and saliva-enhancing properties but also to treat various diseases. NAA containing products are abundantly available on the market as food, cosmetics, medical devices and medicinal products. However, no unambiguous legal product classification is applied for these products. In this study, the different health product classes from a European viewpoint are discussed in relation to the pharmacokinetic and pharmacodynamic properties of the NAAs, their applied dosage and claimed usage.


Assuntos
Amidas/classificação , Qualidade de Produtos para o Consumidor/legislação & jurisprudência , Amidas/farmacologia , Animais , Europa (Continente) , Regulamentação Governamental , Humanos
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