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1.
Proteins ; 92(9): 1070-1084, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38497314

RESUMO

Human islet amyloid polypeptide (amylin or hIAPP) is a 37 residue hormone co-secreted with insulin from ß cells of the pancreas. In patients suffering from type-2 diabetes, amylin self-assembles into amyloid fibrils, ultimately leading to the death of the pancreatic cells. However, a research gap exists in preventing and treating such amyloidosis. Plumbagin, a natural compound, has previously been demonstrated to have inhibitory potential against insulin amyloidosis. Our investigation unveils collapsible regions within hIAPP that, upon collapse, facilitates hydrophobic and pi-pi interactions, ultimately leading to aggregation. Intriguingly plumbagin exhibits the ability to bind these specific collapsible regions, thereby impeding the aforementioned interactions that would otherwise drive hIAPP aggregation. We have used atomistic molecular dynamics approach to determine secondary structural changes. MSM shows metastable states forming native like hIAPP structure in presence of PGN. Our in silico results concur with in vitro results. The ThT assay revealed a striking 50% decrease in fluorescence intensity at a 1:1 ratio of hIAPP to Plumbagin. This finding suggests a significant inhibition of amyloid fibril formation by plumbagin, as ThT fluorescence directly correlates with the presence of these fibrils. Further TEM images revealed disappearance of hIAPP fibrils in plumbagin pre-treated hIAPP samples. Also, we have shown that plumbagin disrupts the intermolecular hydrogen bonding in hIAPP fibrils leading to an increase in the average beta strand spacing, thereby causing disaggregation of pre-formed fibrils demonstrating overall disruption of the aggregation machinery of hIAPP. Our work is the first to report a detailed atomistic simulation of 22 µs for hIAPP. Overall, our studies put plumbagin as a potential candidate for both preventive and therapeutic candidate for hIAPP amyloidosis.


Assuntos
Polipeptídeo Amiloide das Ilhotas Pancreáticas , Simulação de Dinâmica Molecular , Naftoquinonas , Naftoquinonas/química , Naftoquinonas/farmacologia , Polipeptídeo Amiloide das Ilhotas Pancreáticas/química , Polipeptídeo Amiloide das Ilhotas Pancreáticas/metabolismo , Polipeptídeo Amiloide das Ilhotas Pancreáticas/antagonistas & inibidores , Humanos , Cadeias de Markov , Ligação Proteica , Interações Hidrofóbicas e Hidrofílicas , Agregados Proteicos/efeitos dos fármacos , Ligação de Hidrogênio
2.
J Phys Chem B ; 124(9): 1637-1652, 2020 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-32045248

RESUMO

Insulin, which is a small protein hormone consisting of 51 amino acids, rapidly fibrillates under stressogenic conditions. This biotechnological/medical problematic reaction quickly accelerates in the presence of some particles, while there are several other particles that slow down the kinetic process. To address the unexplored demand of the particles that modulate protein fibrillation, we have synthesized two amino-based particles and a chitosan-coated mesoporous silica particle (MS-NH2, MS-3NH2, and MS-chitosan) to investigate insulin fibrillation. While these particles were fairly similar in size, they are differ in their net positive charge and surface hydrophobicity. To monitor the exact role of the hydrophobic interaction between the protein and MS-chitosan during the fibrillation, we have also co- and preincubated insulin with cholesterol and the particles under stressogenic conditions. The results indicate that MS-NH2 and MS-3NH2, due to their high positive charges and lack of surface hydrophobicity, repel the positively charged unfolded insulins at pH 2.0. Moreover, MS-chitosan with 25% surface hydrophobicity stacks partially unfolded insulins to its surface and induces some α-helix to ß-sheet structural transitions to the protein. Consequently, both amino- and chitosan-based particles slow down the kinetics of the fibrillation. We also showed that cholesterol can structurally participate in insulin fibril architecture as a hydrophobic bridge, and extraction of this molecule from the preformed fibrils may disrupt the fibril structure.


Assuntos
Insulina/química , Agregados Proteicos/efeitos dos fármacos , Dióxido de Silício/química , Dióxido de Silício/farmacologia , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Tamanho da Partícula , Porosidade
3.
Protein Pept Lett ; 27(3): 243-250, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31738131

RESUMO

BACKGROUND: Thrombosis represents as the prime contributor to the burden of diseases, worldwide. Conventional anticoagulants for thrombosis therapy have a common bleeding side effect. Bioactive peptides are studied to be an effective alternative for currently available therapeutic drugs. OBJECTIVE: In this study, VITPOR AI peptide, a previously reported coagulation FXIIa inhibitor from Nori (Porphyra yezoensis), was assessed for its inhibitory activity against FXIIa and its in vivo mode of action. METHODS: In vivo efficacy as well as the antithrombotic property of the peptide was evaluated in mice model by ex vivo activated Partial Thromboplastin Time assay, tail transection model and whole blood clotting time. The enzyme kinetics was studied using chromogenic substrate assay. RESULTS: The kinetic behaviour of VITPOR AI showed that the peptide is a competitive inhibitor of FXIIa. Peptide showed significant inhibition of platelet adhesion and aggregation. VITPOR AI exhibited significant antithrombotic activity. Furthermore, ex vivo activated Partial Thromboplastin Time assay revealed that VITPOR AI exhibited potent anticoagulant activity in vivo. Tail bleeding assay revealed that the peptide did not prolong bleeding time in mice even at a higher dose of 5 mg/kg. Cytotoxicity studies of the peptide against human blood leukocytes indicated the safety of the peptide. CONCLUSION: VITPOR AI could be prospected as a potent anticoagulant with Factor XIIa inhibition, antiplatelet aggregation and antithrombotic activity. It was also studied to have no bleeding side effect.


Assuntos
Proteínas de Algas/química , Proteínas Sanguíneas/farmacologia , Peptídeos/farmacologia , Porphyra/metabolismo , Animais , Coagulação Sanguínea/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Feminino , Humanos , Masculino , Camundongos , Tempo de Tromboplastina Parcial , Agregados Proteicos/efeitos dos fármacos
4.
Biochemistry ; 57(18): 2606-2610, 2018 05 08.
Artigo em Inglês | MEDLINE | ID: mdl-29638118

RESUMO

Antimicrobial peptides (AMPs) are a promising alternative to antibiotics for mitigating bacterial infections, in light of increasing bacterial resistance to antibiotics. However, predicting, understanding, and controlling the antibacterial activity of AMPs remain a significant challenge. While peptide intramolecular interactions are known to modulate AMP antimicrobial activity, peptide intermolecular interactions remain elusive in their impact on peptide bioactivity. Herein, we test the relationship between AMP intermolecular interactions and antibacterial efficacy by controlling AMP intermolecular hydrophobic and hydrogen bonding interactions. Molecular dynamics simulations and Gibbs free energy calculations in concert with experimental assays show that increasing intermolecular interactions via interpeptide aggregation increases the energy cost for the peptide to embed into the bacterial cell membrane, which in turn decreases the AMP antibacterial activity. Our findings provide a route for predicting and controlling the antibacterial activity of AMPs against Gram-negative bacteria via reductions of intermolecular AMP interactions.


Assuntos
Peptídeos Catiônicos Antimicrobianos/química , Metabolismo Energético/efeitos dos fármacos , Agregados Proteicos/efeitos dos fármacos , Peptídeos Catiônicos Antimicrobianos/metabolismo , Peptídeos Catiônicos Antimicrobianos/farmacologia , Membrana Celular/química , Membrana Celular/efeitos dos fármacos , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/patogenicidade , Humanos , Simulação de Dinâmica Molecular
5.
J Alzheimers Dis ; 60(s1): S59-S68, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28453488

RESUMO

Alzheimer's disease (AD) is the most common neurodegenerative disorder affecting the elderly population worldwide. Brain inflammation plays a key role in the progression of AD. Deposition of senile plaques in the brain stimulates an inflammatory response with the overexpression of pro-inflammatory mediators, such as the neuroinflammatory cytokine. interleukin-6. Curcumin has been revealed to be a potential agent for treating AD following different neuroprotective mechanisms, such as inhibition of aggregation and decrease in brain inflammation. We synthesized new curcumin derivatives with the aim of providing good anti-aggregation capacity but also improved anti-inflammatory activity. Nine curcumin derivatives were synthesized by etherification and esterification of the aromatic region. From these derivatives, compound 8 exhibited an anti-inflammatory effect similar to curcumin, while compounds 3, 4, and 10 were more potent. Moreover, when the anti-aggregation activity is considered, compounds 3, 4, 5, 6, and 10 showed biological activity in vitro. Compound 4 exhibited a strong anti-aggregation effect higher than curcumin. Monofunctionalized curcumin derivatives showed better bioactivity than difunctionalized compounds. Moreover, the presence of bulky groups in the chemical structure of curcumin derivatives decreased bioactivity.


Assuntos
Peptídeos beta-Amiloides/metabolismo , Anti-Inflamatórios/farmacologia , Curcumina/síntese química , Curcumina/farmacologia , Citocinas/metabolismo , Animais , Anti-Inflamatórios/química , Células Cultivadas , Curcumina/química , Ciclo-Oxigenase 1/metabolismo , Relação Dose-Resposta a Droga , Feminino , Lipopolissacarídeos/toxicidade , Macrófagos/efeitos dos fármacos , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Óxido Nítrico Sintase Tipo II/metabolismo , Agregados Proteicos/efeitos dos fármacos
6.
Eur J Med Chem ; 121: 785-792, 2016 Oct 04.
Artigo em Inglês | MEDLINE | ID: mdl-26608003

RESUMO

The aggregation of a large variety of amyloidogenic proteins is linked to the onset of devastating human disorders. Therefore, there is an urgent need for effective molecules able to modulate the aggregative properties of these polypeptides in their natural environment, in order to prevent, delay or halt the progression of such diseases. On the one hand, the complexity and cost of animal models make them inefficient at early stages of drug discovery, where large chemical libraries are usually screened. On the other hand, in vitro aggregation assays in aqueous solutions hardly reproduce (patho)physiological conditions. In this context, because the formation of insoluble aggregates in bacteria shares mechanistic and functional properties with amyloid self-assembly in higher organisms, they have emerged as a promising system to model aggregation in the cell. Here we show that bacteria provide a powerful and cost-effective system to screen for amyloid inhibitors using fluorescence spectroscopy and flow cytometry, thanks to the ability of the novel red fluorescent ProteoStat dye to detect specifically intracellular amyloid-like aggregates. We validated the approach using the Alzheimer's linked Aß40 and Aß42 peptides and tacrine- and huprine-based aggregation inhibitors. Overall, the present method bears the potential to replace classical in vitro anti-aggregation assays.


Assuntos
Peptídeos beta-Amiloides/química , Avaliação Pré-Clínica de Medicamentos/métodos , Escherichia coli/citologia , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Fragmentos de Peptídeos/química , Agregados Proteicos/efeitos dos fármacos , Amiloide , Análise Custo-Benefício , Avaliação Pré-Clínica de Medicamentos/economia , Escherichia coli/efeitos dos fármacos , Corpos de Inclusão/efeitos dos fármacos , Corpos de Inclusão/metabolismo , Tacrina/química , Tacrina/farmacologia , Fatores de Tempo
8.
Mem. Inst. Oswaldo Cruz ; 110(1): 86-94, 03/02/2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-741617

RESUMO

Trypanosoma cruzi strains from distinct geographic areas show differences in drug resistance and association between parasites genetic and treatment response has been observed. Considering that benznidazole (BZ) can reduce the parasite burden and tissues damage, even in not cured animals and individuals, the goal is to assess the drug response to BZ of T. cruzi II strains isolated from children of the Jequitinhonha Valley, state of Minas Gerais, Brazil, before treatment. Mice infected and treated with BZ in both phases of infection were compared with the untreated and evaluated by fresh blood examination, haemoculture, polymerase chain reaction, conventional (ELISA) and non-conventional (FC-ALTA) serologies. In mice treated in the acute phase, a significant decrease in parasitaemia was observed for all strains. Positive parasitological and/or serological tests in animals treated during the acute and chronic (95.1-100%) phases showed that most of the strains were BZ resistant. However, beneficial effect was demonstrated because significant reduction (p < 0.05%) and/or suppression of parasitaemia was observed in mice infected with all strains (acute phase), associated to reduction/elimination of inflammation and fibrosis for two/eight strains. BZ offered some benefit, even in not cured animals, what suggest that BZ use may be recommended at least for recent chronic infection of the studied region.


Assuntos
Humanos , Descoberta de Drogas , Resíduos Industriais/análise , Nootrópicos/isolamento & purificação , Extratos Vegetais/química , Brotos de Planta/química , Estilbenos/isolamento & purificação , Vitis/química , Agricultura/economia , Peptídeos beta-Amiloides/antagonistas & inibidores , Peptídeos beta-Amiloides/metabolismo , Benzofuranos/análise , Benzofuranos/química , Benzofuranos/economia , Benzofuranos/isolamento & purificação , Cromatografia Líquida de Alta Pressão , França , Resíduos Industriais/economia , Estrutura Molecular , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/economia , Fármacos Neuroprotetores/isolamento & purificação , Fármacos Neuroprotetores/farmacologia , Nootrópicos/química , Nootrópicos/economia , Nootrópicos/farmacologia , Agregação Patológica de Proteínas , Fragmentos de Peptídeos/antagonistas & inibidores , Fragmentos de Peptídeos/metabolismo , Fenóis/química , Fenóis/economia , Extratos Vegetais/economia , Agregados Proteicos/efeitos dos fármacos , Espectrometria de Massas por Ionização por Electrospray , Estereoisomerismo , Estilbenos/análise , Estilbenos/química , Estilbenos/economia , Estilbenos/farmacologia
9.
J Sci Food Agric ; 94(5): 951-4, 2014 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-23929536

RESUMO

BACKGROUND: Viticultural residues from commercial viticultural activities represent a potentially important source of bioactive stilbenes such as resveratrol. The main aim of the present study was therefore to isolate, identify and perform biological assays against amyloid-ß peptide aggregation of original stilbenes from Vitis vinifera shoots. RESULTS: A new resveratrol oligomer, (Z)-cis-miyabenol C (3), was isolated from Vitis vinifera grapevine shoots together with two newly reported oligostilbenes from Vitis vinifera shoots, vitisinol C (1) and (E)-cis-miyabenol C (2), and six known compounds: piceatannol, resveratrol, (E)-ε-viniferin (trans-ε-viniferin), ω-viniferin, vitisinol C and (E)-miyabenol C. The structures of these resveratrol derivatives were established on the basis of detailed spectroscopic analysis including nuclear magnetic resonance experiments. All the newly reported compounds were tested for their anti-aggregative activity against amyloid-ß fibril formation. Vitisinol C was found to exert a significant activity against amyloid-ß aggregation. CONCLUSION: Vitis vinifera grapevine shoots are potentially interesting as a source of new bioactive stilbenes, such as vitisinol C.


Assuntos
Descoberta de Drogas , Resíduos Industriais/análise , Nootrópicos/isolamento & purificação , Extratos Vegetais/química , Brotos de Planta/química , Estilbenos/isolamento & purificação , Vitis/química , Agricultura/economia , Peptídeos beta-Amiloides/antagonistas & inibidores , Peptídeos beta-Amiloides/metabolismo , Benzofuranos/análise , Benzofuranos/química , Benzofuranos/economia , Benzofuranos/isolamento & purificação , Cromatografia Líquida de Alta Pressão , França , Humanos , Resíduos Industriais/economia , Estrutura Molecular , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/economia , Fármacos Neuroprotetores/isolamento & purificação , Fármacos Neuroprotetores/farmacologia , Nootrópicos/química , Nootrópicos/economia , Nootrópicos/farmacologia , Fragmentos de Peptídeos/antagonistas & inibidores , Fragmentos de Peptídeos/metabolismo , Fenóis/química , Fenóis/economia , Extratos Vegetais/economia , Agregados Proteicos/efeitos dos fármacos , Agregação Patológica de Proteínas , Espectrometria de Massas por Ionização por Electrospray , Estereoisomerismo , Estilbenos/análise , Estilbenos/química , Estilbenos/economia , Estilbenos/farmacologia , Estilbestrois
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