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1.
J Enzyme Inhib Med Chem ; 36(1): 727-736, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33685335

RESUMEN

The novel coronavirus disease COVID-19, caused by the virus SARS CoV-2, has exerted a significant unprecedented economic and medical crisis, in addition to its impact on the daily life and health care systems all over the world. Regrettably, no vaccines or drugs are currently available for this new critical emerging human disease. Joining the global fight against COVID-19, in this study we aim at identifying a potential novel inhibitor for SARS COV-2 2'-O-methyltransferase (nsp16) which is one of the most attractive targets in the virus life cycle, responsible for the viral RNA protection via a cap formation process. Firstly, nsp16 enzyme bound to Sinefungin was retrieved from the protein data bank (PDB ID: 6WKQ), then, a 3D pharmacophore model was constructed to be applied to screen 48 Million drug-like compounds of the Zinc database. This resulted in only 24 compounds which were subsequently docked into the enzyme. The best four score-ordered hits from the docking outcome exhibited better scores compared to Sinefungin. Finally, three molecular dynamics (MD) simulation experiments for 150 ns were carried out as a refinement step for our proposed approach. The MD and MM-PBSA outputs revealed compound 11 as the best potential nsp16 inhibitor herein identified, as it displayed a better stability and average binding free energy for the ligand-enzyme complex compared to Sinefungin.


Asunto(s)
Antivirales/química , Inhibidores Enzimáticos/química , Proteínas no Estructurales Virales/química , Adenosina/análogos & derivados , Adenosina/química , Adenosina/metabolismo , Antivirales/metabolismo , Sitios de Unión , Cristalografía por Rayos X , Bases de Datos Farmacéuticas , Bases de Datos de Proteínas , Estabilidad de Medicamentos , Inhibidores Enzimáticos/metabolismo , Ensayos Analíticos de Alto Rendimiento , Humanos , Cinética , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Unión Proteica , Conformación Proteica en Hélice alfa , Conformación Proteica en Lámina beta , Dominios y Motivos de Interacción de Proteínas , Termodinámica , Proteínas no Estructurales Virales/antagonistas & inhibidores
2.
Viruses ; 13(2)2021 02 17.
Artículo en Inglés | MEDLINE | ID: mdl-33671334

RESUMEN

MASP-2, mannose-binding protein-associated serine protease 2, is a key enzyme in the lectin pathway of complement activation. Hyperactivation of this protein by human coronaviruses SARS-CoV, MERS-CoV and SARS-CoV-2 has been found to contribute to aberrant complement activation in patients, leading to aggravated lung injury with potentially fatal consequences. This hyperactivation is triggered in the lungs through a conserved, direct interaction between MASP-2 and coronavirus nucleocapsid (N) proteins. Blocking this interaction with monoclonal antibodies and interfering directly with the catalytic activity of MASP-2, have been found to alleviate coronavirus-induced lung injury both in vitro and in vivo. In this study, a virtual library of 8736 licensed drugs and clinical agents has been screened in silico according to two parallel strategies. The first strategy aims at identifying direct inhibitors of MASP-2 catalytic activity, while the second strategy focusses on finding protein-protein interaction inhibitors (PPIs) of MASP-2 and coronaviral N proteins. Such agents could represent promising support treatment options to prevent lung injury and reduce mortality rates of infections caused by both present and future-emerging coronaviruses. Forty-six drug repurposing candidates were purchased and, for the ones selected as potential direct inhibitors of MASP-2, a preliminary in vitro assay was conducted to assess their interference with the lectin pathway of complement activation. Some of the tested agents displayed a dose-response inhibitory activity of the lectin pathway, potentially providing the basis for a viable support strategy to prevent the severe complications of coronavirus infections.


Asunto(s)
Inhibidores Enzimáticos/química , Serina Proteasas Asociadas a la Proteína de Unión a la Manosa/metabolismo , Unión Proteica/efectos de los fármacos , Infecciones por Coronavirus/tratamiento farmacológico , /metabolismo , Reposicionamiento de Medicamentos , Humanos , Relación Estructura-Actividad
3.
Molecules ; 26(4)2021 Feb 11.
Artículo en Inglés | MEDLINE | ID: mdl-33670416

RESUMEN

Growing scientific evidence indicates that Achillea biebersteinii is a valuable source of active ingredients with potential cosmetic applications. However, the data on its composition and pharmacological properties are still insufficient. This study aims to optimize the extraction procedure of the plant material, evaluate its phytochemical composition, and compare anti-tyrosinase potential of A. biebersteinii extracts obtained by various methods. In order to identify compounds responsible for the tyrosinase inhibitory activity of A. biebersteinii, the most active anti-tyrosinase extract was fractionated by column chromatography. The fractions were examined for their skin lightening potential by mushroom and murine tyrosinase inhibitory assays and melanin release assay. HPLC-ESI-Q-TOF-MS/MS analysis of the total extract revealed the presence of several phenolic acids, flavonoids, flavonoid glucosides, and carboxylic acid. Among them, fraxetin-8-O-glucoside, quercetin-O-glucopyranose, schaftoside/isoschaftoside, gmelinin B, 1,3-dicaffeoylquinic acid (1,3-DCQA), and ferulic acid were found in the fractions with the highest skin lightening potential. Based on obtained qualitative and quantitative analysis of the fractions, it was assumed that the caffeoylquinic acid derivatives and dicaffeoylquinic acid derivatives are more likely responsible for mushroom tyrosinase inhibitory activity of A. biebersteinii extracts and fractions. Ferulic acid was proposed as the most active murine tyrosinase inhibitor, responsible also for the reduced melanin release from B16F10 murine melanoma cells.


Asunto(s)
Achillea/química , Agaricales/enzimología , Inhibidores Enzimáticos/aislamiento & purificación , Monofenol Monooxigenasa/química , Animales , Antioxidantes/química , Inhibidores Enzimáticos/química , Flavonoides/química , Ratones , Monofenol Monooxigenasa/antagonistas & inhibidores , Fitoquímicos/química , Extractos Vegetales/química , Espectrometría de Masas en Tándem
4.
Molecules ; 26(4)2021 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-33673088

RESUMEN

The dysregulation of Notch signaling is associated with a wide variety of different human cancers. Notch signaling activation mostly relies on the activity of the γ-secretase enzyme that cleaves the Notch receptors and releases the active intracellular domain. It is well-documented that γ-secretase inhibitors (GSIs) block the Notch activity, mainly by inhibiting the oncogenic activity of this pathway. To date, several GSIs have been introduced clinically for the treatment of various diseases, such as Alzheimer's disease and various cancers, and their impacts on Notch inhibition have been found to be promising. Therefore, GSIs are of great interest for cancer therapy. The objective of this review is to provide a systematic review of in vitro and in vivo studies for investigating the effect of GSIs on various cancer stem cells (CSCs), mainly by modulation of the Notch signaling pathway. Various scholarly electronic databases were searched and relevant studies published in the English language were collected up to February 2020. Herein, we conclude that GSIs can be potential candidates for CSC-targeting therapy. The outcome of our study also indicates that GSIs in combination with anticancer drugs have a greater inhibitory effect on CSCs.


Asunto(s)
Secretasas de la Proteína Precursora del Amiloide/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Neoplasias/tratamiento farmacológico , Células Madre Neoplásicas/efectos de los fármacos , Secretasas de la Proteína Precursora del Amiloide/genética , Antineoplásicos/química , Antineoplásicos/uso terapéutico , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Inhibidores Enzimáticos/uso terapéutico , Humanos , Células Madre Neoplásicas/patología , Receptores Notch/antagonistas & inhibidores , Receptores Notch/genética
5.
Molecules ; 26(4)2021 Feb 22.
Artículo en Inglés | MEDLINE | ID: mdl-33671811

RESUMEN

Marine algae are a promising source of potent bioactive agents against oxidative stress, diabetes, and inflammation. However, the possible therapeutic effects of many algal metabolites have not been exploited yet. In this regard, we explored the therapeutic potential of Enteromorpha intestinalis extracts obtained from methanol, ethanol, and hexane, in contrasting oxidative stress. The total phenolic (TPC) and flavonoids (TFC) content were quantified in all extracts, with ethanol yielding the best values (about 60 and 625 mg of gallic acid and rutin equivalents per gram of extract, respectively). Their antioxidant potential was also assessed through DPPH•, hydroxyl radical, hydrogen peroxide, and superoxide anion scavenging assays, showing a concentration-dependent activity which was greater in the extracts from protic and more polar solvents. The α-amylase and α-glucosidase activities were estimated for checking the antidiabetic capacity, with IC50 values of about 3.8 µg/mL for the methanolic extract, almost as low as those obtained with acarbose (about 2.8 and 3.3 µg/mL, respectively). The same extract also showed remarkable anti-inflammatory effect, as determined by hemolysis, protein denaturation, proteinase and lipoxygenase activity assays, with respectable IC50 values (about 11, 4, 6, and 5 µg/mL, respectively), also in comparison to commercially used drugs, such as acetylsalicylic acid.


Asunto(s)
Antiinflamatorios/farmacología , Antioxidantes/farmacología , Inhibidores Enzimáticos/farmacología , Hipoglucemiantes/farmacología , Extractos Vegetales/farmacología , Ulva/química , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Compuestos de Bifenilo/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/aislamiento & purificación , Eritrocitos/efectos de los fármacos , Eritrocitos/metabolismo , Flavonoides/química , Flavonoides/aislamiento & purificación , Flavonoides/farmacología , Humanos , Radical Hidroxilo/antagonistas & inhibidores , Hipoglucemiantes/química , Hipoglucemiantes/aislamiento & purificación , Lipooxigenasas/metabolismo , Masculino , Péptido Hidrolasas/metabolismo , Fenoles/química , Fenoles/aislamiento & purificación , Fenoles/farmacología , Picratos/antagonistas & inhibidores , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Superóxidos/antagonistas & inhibidores
6.
Molecules ; 26(4)2021 Feb 13.
Artículo en Inglés | MEDLINE | ID: mdl-33668468

RESUMEN

Nicotinamide-N-methyltransferase (NNMT) is a cytosolic enzyme catalyzing the transfer of a methyl group from S-adenosyl-methionine (SAM) to nicotinamide (Nam). It is expressed in many tissues including the liver, adipose tissue, and skeletal muscle. Its expression in several cancer cell lines has been widely discussed in the literature, and recent work established a link between NNMT expression and metabolic diseases. Here we describe our approach to identify potent small molecule inhibitors of NNMT featuring different binding modes as elucidated by X-ray crystallographic studies.


Asunto(s)
Inhibidores Enzimáticos/uso terapéutico , Enfermedades Metabólicas/tratamiento farmacológico , Enfermedades Metabólicas/enzimología , Nicotinamida N-Metiltransferasa/antagonistas & inhibidores , Animales , Sitios de Unión , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Ensayos Analíticos de Alto Rendimiento , Humanos , Ligandos , Ratones , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/enzimología , Modelos Moleculares , Niacinamida/metabolismo , Nicotinamida N-Metiltransferasa/metabolismo , Ratas , Especificidad por Sustrato/efectos de los fármacos
7.
Food Chem ; 351: 129338, 2021 Jul 30.
Artículo en Inglés | MEDLINE | ID: mdl-33647700

RESUMEN

Hyperstable cysteine-rich peptides (CRPs) represent an underexplored superfamily of bioactives in functional foods. An example is wolfberry of the Lycium barbarum family. Previously, we discovered a CRP, designated α-lybatide, from L. barbarum bark. Herein, we report the discovery of ß-lybatide, a novel carboxypeptidase inhibitor belonging to a different CRP family from the wolfberry plant. Proteomic and transcriptomic analyses showed that ß-lybatide contains 36 amino acids with six cysteine residues. NMR spectroscopy revealed that ß-lybatide displays a knottin-like structure that renders it highly resistant to thermal, chemical and enzymatic degradation, conditions important for keeping its structural integrity in gastrointestinal tract. Biochemical assays showed that ß-lybatide is a potent carboxypeptidase inhibitor which could contribute to the wolfberry biological activities. Bioinformatics analysis revealed an additional 49 ß-lybatide-like plant carboxypeptidase inhibitors. Together, our results show that ß-lybatide is the first and the smallest plant-derived hyperstable carboxypeptidase inhibitor discovered from a functional food.


Asunto(s)
Carboxipeptidasas/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Lycium/química , Péptidos/farmacología , Cisteína , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/metabolismo , Lycium/metabolismo , Péptidos/química , Péptidos/metabolismo , Proteómica
8.
Nat Commun ; 12(1): 1782, 2021 03 19.
Artículo en Inglés | MEDLINE | ID: mdl-33741963

RESUMEN

Pharmacological inhibition of vacuolar-type H+-ATPase (V-ATPase) by its specific inhibitor can abrogate tumor metastasis, prevent autophagy, and reduce cellular signaling responses. Bafilomycin A1, a member of macrolide antibiotics and an autophagy inhibitor, serves as a specific and potent V-ATPases inhibitor. Although there are many V-ATPase structures reported, the molecular basis of specific inhibitors on V-ATPase remains unknown. Here, we report the cryo-EM structure of bafilomycin A1 bound intact bovine V-ATPase at an overall resolution of 3.6-Å. The structure reveals six bafilomycin A1 molecules bound to the c-ring. One bafilomycin A1 molecule engages with two c subunits and disrupts the interactions between the c-ring and subunit a, thereby preventing proton translocation. Structural and sequence analyses demonstrate that the bafilomycin A1-binding residues are conserved in yeast and mammalian species and the 7'-hydroxyl group of bafilomycin A1 acts as a unique feature recognized by subunit c.


Asunto(s)
Macrólidos/farmacología , ATPasas de Translocación de Protón Vacuolares/antagonistas & inhibidores , Secuencia de Aminoácidos , Animales , Sitios de Unión , Biocatálisis/efectos de los fármacos , Bovinos , Microscopía por Crioelectrón , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/metabolismo , Inhibidores Enzimáticos/farmacología , Macrólidos/química , Macrólidos/metabolismo , Modelos Moleculares , Estructura Molecular , Unión Proteica , Dominios Proteicos , Homología de Secuencia de Aminoácido , ATPasas de Translocación de Protón Vacuolares/química , ATPasas de Translocación de Protón Vacuolares/ultraestructura
9.
Nat Struct Mol Biol ; 28(3): 319-325, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33674802

RESUMEN

The COVID-19 pandemic caused by nonstop infections of SARS-CoV-2 has continued to ravage many countries worldwide. Here we report that suramin, a 100-year-old drug, is a potent inhibitor of the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) and acts by blocking the binding of RNA to the enzyme. In biochemical assays, suramin and its derivatives are at least 20-fold more potent than remdesivir, the currently approved nucleotide drug for treatment of COVID-19. The 2.6 Å cryo-electron microscopy structure of the viral RdRp bound to suramin reveals two binding sites. One site directly blocks the binding of the RNA template strand and the other site clashes with the RNA primer strand near the RdRp catalytic site, thus inhibiting RdRp activity. Suramin blocks viral replication in Vero E6 cells, although the reasons underlying this effect are likely various. Our results provide a structural mechanism for a nonnucleotide inhibitor of the SARS-CoV-2 RdRp.


Asunto(s)
Antivirales/farmacología , /química , Inhibidores Enzimáticos/farmacología , Suramina/farmacología , Animales , Antivirales/química , Antivirales/metabolismo , Sitios de Unión , Dominio Catalítico , Chlorocebus aethiops , Microscopía por Crioelectrón , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/metabolismo , Conformación Proteica , ARN Viral/química , ARN Viral/metabolismo , Suramina/química , Suramina/metabolismo , Células Vero , Replicación Viral/efectos de los fármacos
10.
Commun Biol ; 4(1): 193, 2021 02 09.
Artículo en Inglés | MEDLINE | ID: mdl-33564093

RESUMEN

SARS-CoV-2 Nsp15 is a uridine-specific endoribonuclease with C-terminal catalytic domain belonging to the EndoU family that is highly conserved in coronaviruses. As endoribonuclease activity seems to be responsible for the interference with the innate immune response, Nsp15 emerges as an attractive target for therapeutic intervention. Here we report the first structures with bound nucleotides and show how the enzyme specifically recognizes uridine moiety. In addition to a uridine site we present evidence for a second base binding site that can accommodate any base. The structure with a transition state analog, uridine vanadate, confirms interactions key to catalytic mechanisms. In the presence of manganese ions, the enzyme cleaves unpaired RNAs. This acquired knowledge was instrumental in identifying Tipiracil, an FDA approved drug that is used in the treatment of colorectal cancer, as a potential anti-COVID-19 drug. Using crystallography, biochemical, and whole-cell assays, we demonstrate that Tipiracil inhibits SARS-CoV-2 Nsp15 by interacting with the uridine binding pocket in the enzyme's active site. Our findings provide new insights for the development of uracil scaffold-based drugs.


Asunto(s)
Antivirales/farmacología , /virología , Endorribonucleasas/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Pirrolidinas/farmacología , /enzimología , Timina/farmacología , Proteínas no Estructurales Virales/antagonistas & inhibidores , Células A549 , Antivirales/química , Antivirales/farmacocinética , Dominio Catalítico , Cristalografía por Rayos X , Endorribonucleasas/química , Endorribonucleasas/metabolismo , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacocinética , Humanos , Ligandos , Modelos Moleculares , Conformación Proteica , Pirrolidinas/química , Pirrolidinas/farmacocinética , Timina/química , Timina/farmacocinética , Uridina/metabolismo , Proteínas no Estructurales Virales/química , Proteínas no Estructurales Virales/metabolismo
11.
SAR QSAR Environ Res ; 32(2): 85-110, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33517778

RESUMEN

Tyrosinase is a key rate-limiting enzyme in the process of melanin synthesis, which is closely related to human pigmentation disorders. Tyrosinase inhibitors can down-regulate tyrosinase to effectively reduce melanin synthesis. In this work, we conducted structure-activity relationship (SAR) study on 1097 diverse mushroom tyrosinase inhibitors. We applied five kinds of machine learning methods to develop 15 classification models. Model 5B built by fully connected neural networks and ECFP4 fingerprints achieved the highest prediction accuracy of 91.36% and Matthews correlation coefficient (MCC) of 0.81 on the test set. The applicability domains (AD) of classification models were defined by d S T D - P R O method. Moreover, we clustered the 1097 inhibitors into eight subsets by K-Means to figure out inhibitors' structural features. In addition, 10 quantitative structure-activity relationship (QSAR) models were constructed by four machine learning methods based on 813 inhibitors. Model 6 J, the best QSAR model, was developed by fully connected neural networks with 50 RDKit descriptors. It resulted in a coefficient of determination (r 2) of 0.770 and a root mean squared error (RMSE) of 0.482 on the test set. The AD of Model 6 J was visualized by Williams plot. The models built in this study can be obtained from the authors.


Asunto(s)
Inhibidores Enzimáticos/química , Aprendizaje Automático , Monofenol Monooxigenasa/antagonistas & inhibidores , Redes Neurales de la Computación , Relación Estructura-Actividad , Relación Estructura-Actividad Cuantitativa
12.
Proc Natl Acad Sci U S A ; 118(7)2021 02 16.
Artículo en Inglés | MEDLINE | ID: mdl-33526596

RESUMEN

The RNA polymerase inhibitor favipiravir is currently in clinical trials as a treatment for infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), despite limited information about the molecular basis for its activity. Here we report the structure of favipiravir ribonucleoside triphosphate (favipiravir-RTP) in complex with the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) bound to a template:primer RNA duplex, determined by electron cryomicroscopy (cryoEM) to a resolution of 2.5 Å. The structure shows clear evidence for the inhibitor at the catalytic site of the enzyme, and resolves the conformation of key side chains and ions surrounding the binding pocket. Polymerase activity assays indicate that the inhibitor is weakly incorporated into the RNA primer strand, and suppresses RNA replication in the presence of natural nucleotides. The structure reveals an unusual, nonproductive binding mode of favipiravir-RTP at the catalytic site of SARS-CoV-2 RdRp, which explains its low rate of incorporation into the RNA primer strand. Together, these findings inform current and future efforts to develop polymerase inhibitors for SARS coronaviruses.


Asunto(s)
Amidas/farmacología , Inhibidores Enzimáticos/farmacología , Pirazinas/farmacología , /ultraestructura , Amidas/química , /química , Microscopía por Crioelectrón/métodos , Inhibidores Enzimáticos/química , Pirazinas/química , Ribonucleótidos/química , /enzimología , Imagen Individual de Molécula/métodos
13.
Eur J Med Chem ; 213: 113201, 2021 Mar 05.
Artículo en Inglés | MEDLINE | ID: mdl-33524687

RESUMEN

The widespread nature of several viruses is greatly credited to their rapidly altering RNA genomes that enable the infection to persist despite challenges presented by host cells. Within the RNA genome of infections is RNA-dependent RNA polymerase (RdRp), which is an essential enzyme that helps in RNA synthesis by catalysing the RNA template-dependent development of phosphodiester bonds. Therefore, RdRp is an important therapeutic target in RNA virus-caused diseases, including SARS-CoV-2. In this review, we describe the promising RdRp inhibitors that have been launched or are currently in clinical studies for the treatment of RNA virus infections. Structurally, nucleoside inhibitors (NIs) bind to the RdRp protein at the enzyme active site, and nonnucleoside inhibitors (NNIs) bind to the RdRp protein at allosteric sites. By reviewing these inhibitors, more precise guidelines for the development of more promising anti-RNA virus drugs should be set, and due to the current health emergency, they will eventually be used for COVID-19 treatment.


Asunto(s)
Antivirales/uso terapéutico , /antagonistas & inhibidores , Reposicionamiento de Medicamentos , Inhibidores Enzimáticos/uso terapéutico , Animales , Antivirales/química , Inhibidores Enzimáticos/química , Humanos , Nucleósidos/química , Nucleósidos/uso terapéutico , Pandemias , /enzimología
14.
Food Chem ; 347: 129068, 2021 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-33486365

RESUMEN

This study aimed to isolate and investigate the potential of the peptide alanine-cysteine-glutamic acid-cysteine-aspartic acid (ACECD), a novel xanthine oxidase inhibitory (XODI) peptide derived from Skipjack tuna hydrolysate (HS). Ultrafiltration membranes were used to obtain HS-based peptides as successive ultrafiltration fractions (of decreasing molecular weight) of UF-1, UF-2, UF-3, and UF-4. Their antioxidant and xanthine oxidase (XOD) inhibitory activities were determined and further characterized by affinity-ultrafiltration coupled with HPLC-MALDI-TOF/TOF-MS and in silico techniques. The results showed that peptides with a molecular weight (MW) cutoff of 600-1000 Da (UF-2) exhibited the highest antioxidant and XODI activities. A novel XODI peptide (ACECD) was identified with an IC50 value of 13.40 mmol/L, which decreased by 21.24% and 51.40% compared to those of UF-2 and HS, respectively. Molecular docking indicated that ACECD inserted into the active center of Mo atoms in XOD, which led to competitive attachment with XOD and caused inhibition. The study findings indicated that the ACECD peptide could be useful as a safe XODI substance to alleviate hyperuricemia.


Asunto(s)
Inhibidores Enzimáticos/química , Péptidos/química , Atún/metabolismo , Xantina Oxidasa/antagonistas & inhibidores , Secuencia de Aminoácidos , Animales , Antioxidantes/química , Sitios de Unión , Cromatografía Líquida de Alta Presión , Inhibidores Enzimáticos/aislamiento & purificación , Inhibidores Enzimáticos/metabolismo , Simulación del Acoplamiento Molecular , Péptidos/aislamiento & purificación , Péptidos/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Ultrafiltración , Xantina Oxidasa/metabolismo
15.
Arch Biochem Biophys ; 700: 108771, 2021 03 30.
Artículo en Inglés | MEDLINE | ID: mdl-33485847

RESUMEN

In the current study, a structure-based virtual screening paradigm was used to screen a small molecular database against the Non-structural protein 15 (Nsp15) endoribonuclease of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). The SARS-CoV-2 is the causative agent of the recent outbreak of coronavirus disease 2019 (COVID-19) which left the entire world locked down inside the home. A multi-step molecular docking study was performed against antiviral specific compounds (~8722) collected from the Asinex antiviral database. The less or non-interacting molecules were wiped out sequentially in the molecular docking. Further, MM-GBSA based binding free energy was estimated for 26 compounds which shows a high affinity towards the Nsp15. The drug-likeness and pharmacokinetic parameters of all 26 compounds were explored, and five molecules were found to have an acceptable pharmacokinetic profile. Overall, the Glide-XP docking score and Prime-MM-GBSA binding free energy of the selected molecules were explained strong interaction potentiality towards the Nsp15 endoribonuclease. The dynamic behavior of each molecule with Nsp15 was assessed using conventional molecular dynamics (MD) simulation. The MD simulation information was strongly favors the Nsp15 and each identified ligand stability in dynamic condition. Finally, from the MD simulation trajectories, the binding free energy was estimated using the MM-PBSA method. Hence, the proposed final five molecules might be considered as potential Nsp15 modulators for SARS-CoV-2 inhibition.


Asunto(s)
Antivirales/farmacología , /virología , Endorribonucleasas/antagonistas & inhibidores , /enzimología , Proteínas no Estructurales Virales/antagonistas & inhibidores , Antivirales/química , Antivirales/farmacocinética , Bases de Datos de Compuestos Químicos , Evaluación Preclínica de Medicamentos , Endorribonucleasas/química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacocinética , Inhibidores Enzimáticos/farmacología , Humanos , Técnicas In Vitro , Ligandos , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Estructura Molecular , Interfaz Usuario-Computador , Proteínas no Estructurales Virales/química
16.
Molecules ; 26(2)2021 Jan 17.
Artículo en Inglés | MEDLINE | ID: mdl-33477281

RESUMEN

A clear gap with respect to the potential biological properties of wheat flavonoids exists in the available literature. This information is crucial for breeding programs aiming to produce new varieties presenting improved health benefits. Accordingly, advanced breeding lines of whole durum wheat were evaluated in this contribution. The highest recovery of phenolics was achieved using aqueous acetone (50:50, v/v), as verified by multi-response optimization, thus showing that phenolics could be largely underestimated by employing an inappropriate extraction. The concentration of derivatives of apigenin, the main phenolics present, ranged from 63.5 to 80.7%, as evaluated by LC-ESI-QTOF-MS. Phenolics from the breeding line 98 exhibited the highest ability in scavenging peroxyl radicals, reducing power as well as in terms of inhibition of pancreatic lipase activity, a key enzyme regulating the absorption of triacylglycerols. In contrast, none of the samples exhibited a significant anti-diabetic potential. Despite their high concentration compared to that of phenolic acids, results of this work do not support a significant antioxidant and pancreatic lipase inhibitory effect of durum wheat flavonoids. Therefore, breeding programs and animal and/or human trials related to the effect of durum wheat flavonoids on oxidative stress and absorption of triacylglycerols are discouraged at this point.


Asunto(s)
Antioxidantes/química , Inhibidores Enzimáticos/química , Flavonoides/química , Triticum/química , Animales , Humanos , Estudios Prospectivos
17.
Molecules ; 26(2)2021 Jan 10.
Artículo en Inglés | MEDLINE | ID: mdl-33435264

RESUMEN

Diabetes mellitus (DM) is a complex disease which currently affects more than 460 million people and is one of the leading cause of death worldwide. Its development implies numerous metabolic dysfunctions and the onset of hyperglycaemia-induced chronic complications. Multiple ligands can be rationally designed for the treatment of multifactorial diseases, such as DM, with the precise aim of simultaneously controlling multiple pathogenic mechanisms related to the disease and providing a more effective and safer therapeutic treatment compared to combinations of selective drugs. Starting from our previous findings that highlighted the possibility to target both aldose reductase (AR) and protein tyrosine phosphatase 1B (PTP1B), two enzymes strictly implicated in the development of DM and its complications, we synthesised 3-(5-arylidene-4-oxothiazolidin-3-yl)propanoic acids and analogous 2-butenoic acid derivatives, with the aim of balancing the effectiveness of dual AR/PTP1B inhibitors which we had identified as designed multiple ligands (DMLs). Out of the tested compounds, 4f exhibited well-balanced AR/PTP1B inhibitory effects at low micromolar concentrations, along with interesting insulin-sensitizing activity in murine C2C12 cell cultures. The SARs here highlighted along with their rationalization by in silico docking experiments into both target enzymes provide further insights into this class of inhibitors for their development as potential DML antidiabetic candidates.


Asunto(s)
Aldehído Reductasa/antagonistas & inhibidores , Diabetes Mellitus/tratamiento farmacológico , Inhibidores Enzimáticos , Hipoglucemiantes , Proteína Tirosina Fosfatasa no Receptora Tipo 1/antagonistas & inhibidores , Aldehído Reductasa/metabolismo , Animales , Diabetes Mellitus/enzimología , Evaluación Preclínica de Medicamentos , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Células Hep G2 , Humanos , Hipoglucemiantes/química , Hipoglucemiantes/farmacología , Ligandos , Ratones , Proteína Tirosina Fosfatasa no Receptora Tipo 1/metabolismo , Relación Estructura-Actividad
18.
Food Chem ; 348: 129100, 2021 Jun 30.
Artículo en Inglés | MEDLINE | ID: mdl-33515954

RESUMEN

Salicylic acid is generally considered to combine with polyphenol oxidase (PPO) to inhibit activity and enzymatic browning, while its acidification effect on PPO activity was usually neglected. In this study, the inhibitory mechanism of salicylic acid on PPO was examined from acidification and binding effects by altering the buffer conditions. As the buffer concentration increased, contribution of acidification decreased while the binding effect became more predominant. Salicylic acid exhibited competitive inhibition on PPO, inducing the changes in secondary structure with a reduction in α-helix. Molecular docking results showed that salicylic acid interacted with residues HIS61, HIS85, HIS259, HIS263 and VAL283 through hydrogen bond and hydrophobic interaction. Furthermore, acidic pH enhanced the binding of salicylic acid to PPO with lower binding energy, additional hydrogen bond and electrostatic interactions. Therefore, both acidification and binding effects were important for salicylic acid on PPO inhibition and enzymatic browning control in fruit and vegetables.


Asunto(s)
Catecol Oxidasa/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Ácido Salicílico/química , Ácido Salicílico/farmacología , Frutas/química , Concentración de Iones de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Oxidación-Reducción , Verduras/química
19.
Life Sci ; 271: 119115, 2021 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-33515565

RESUMEN

Human pancreatic lipase (triacylglycerol acyl hydrolase EC3.1.1.3) is the most widely studied member of the human lipase superfamily related to carboxyl esterase. It is secreted from the acinar cell of pancreas and has strong preference for triacylglycerides over cholesterol esters, phospholipids, and galactolipids. Apart from the hydrolysis of triacylglycerides, pancreatic lipase may cause the hydrolysis of retinyl esters in vivo. So, it is very much evidenced that pancreatic lipase with its cofactor colipase has prominent role in efficient digestion of dietary fat. Hence, the modulation of human pancreatic lipase may represent a new insight in the discovery of a number of therapeutics that can inhibit the absorption of fat in body and can be used in obesity and other related metabolic disorders. Even, the only Food and drug administration (FDA) approved antiobesity drug, orlistat, is also an inhibitor of pancreatic lipase. This review summarizes studies about structure, mechanistic approach of pancreatic lipase enzyme while emphasizing on the various synthetic pancreatic lipase inhibitors with their structure activity relationship (SAR).


Asunto(s)
Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Lipasa/antagonistas & inhibidores , Lipasa/química , Páncreas/enzimología , Animales , Fármacos Antiobesidad/farmacología , Grasas de la Dieta/antagonistas & inhibidores , Grasas de la Dieta/metabolismo , Humanos , Lipasa/metabolismo , Páncreas/efectos de los fármacos , Estructura Secundaria de Proteína , Relación Estructura-Actividad
20.
Nat Commun ; 12(1): 121, 2021 01 05.
Artículo en Inglés | MEDLINE | ID: mdl-33402676

RESUMEN

p97, also known as valosin-containing protein (VCP) or Cdc48, plays a central role in cellular protein homeostasis. Human p97 mutations are associated with several neurodegenerative diseases. Targeting p97 and its cofactors is a strategy for cancer drug development. Despite significant structural insights into the fungal homolog Cdc48, little is known about how human p97 interacts with its cofactors. Recently, the anti-alcohol abuse drug disulfiram was found to target cancer through Npl4, a cofactor of p97, but the molecular mechanism remains elusive. Here, using single-particle cryo-electron microscopy (cryo-EM), we uncovered three Npl4 conformational states in complex with human p97 before ATP hydrolysis. The motion of Npl4 results from its zinc finger motifs interacting with the N domain of p97, which is essential for the unfolding activity of p97. In vitro and cell-based assays showed that the disulfiram derivative bis-(diethyldithiocarbamate)-copper (CuET) can bypass the copper transporter system and inhibit the function of p97 in the cytoplasm by releasing cupric ions under oxidative conditions, which disrupt the zinc finger motifs of Npl4, locking the essential conformational switch of the complex.


Asunto(s)
Coenzimas/química , Ditiocarba/análogos & derivados , Péptidos y Proteínas de Señalización Intracelular/química , Proteínas Nucleares/química , Compuestos Organometálicos/química , Ubiquitina/química , Proteína que Contiene Valosina/química , Adenosina Trifosfato/análogos & derivados , Adenosina Trifosfato/química , Adenosina Trifosfato/metabolismo , Sitios de Unión , Clonación Molecular , Coenzimas/genética , Coenzimas/metabolismo , Microscopía por Crioelectrón , Disulfiram/química , Disulfiram/metabolismo , Ditiocarba/química , Ditiocarba/metabolismo , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Expresión Génica , Vectores Genéticos/química , Vectores Genéticos/metabolismo , Humanos , Péptidos y Proteínas de Señalización Intracelular/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Modelos Moleculares , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Compuestos Organometálicos/metabolismo , Unión Proteica , Conformación Proteica , Dominios y Motivos de Interacción de Proteínas , Desplegamiento Proteico , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidad por Sustrato , Ubiquitina/genética , Ubiquitina/metabolismo , Proteína que Contiene Valosina/antagonistas & inhibidores , Proteína que Contiene Valosina/genética , Proteína que Contiene Valosina/metabolismo , Dedos de Zinc
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