Preparation and characterization of anticancer niosomal withaferin–A formulation for improved delivery to cancer cells: In vitro, in vivo, and in silico evaluation

AbstractWithaferin-A is a withanolide, predominantly present in Ashwagandha (Withania somnifera). It has been shown to possess anticancer activity in a variety of human cancer cells in vitro and in vivo. Molecular mechanism of such cytotoxicity has not yet been completely understood. Withaferin-A and Withanone were earlier shown to activate p53 tumor suppressor and oxidative stress pathways in cancer cells. 2,3-dihydro-3β-methoxy analogue of Withaferin-A (3βmWi-A) was shown to lack cytotoxicity and well tolerated at higher concentrations. It, on the other hand, protected normal cells against oxidative, chemical and UV stresses through induction of anti-stress and pro-survival signaling. We, in the present study, investigated the effect of Wi-A and 3βmWi-A on cell migration and metastasis signaling. Whereas Wi-A binds to vimentin and heterogeneous nuclear ribonucleoprotein K (hnRNP-K) with high efficacy and downregulates its effector proteins, MMPs and VEGF, involved in cancer cell metastasis, 3βmWi-A was ineffective. Consistently, Wi-A, and not 3βmWi-A, caused reduction in cytoskeleton proteins (Vimentin, N-Cadherin) and active protease (u-PA) that are essential for three key steps of cancer cell metastasis (EMT, increase in cell migration and invasion).

Download Full-text

In-silico structural, functional and immunogenic characterization of Taenia solium TS14 protein

Indian Journal of Animal Research ◽

10.18805/ijar.b-3313 ◽

2017 ◽

Author(s):

Chitra Joshi ◽

Siddharth Gautam

Keyword(s):

Amino Acid ◽

In Silico ◽

Mutational Analysis ◽

Solvent Accessibility ◽

Taenia Solium ◽

Secretory Protein ◽

Single Amino Acid

TS14, a Cysticercosis cellulosae derived protein, has been exploited for immunodiagnosis of cysticercosis in humans and pigs. However, the information on structure, function, stability and immunogenicity of TS14 derived from different isolates is primarily lacking. The present study deals with in-silico characterization of six TS14 isolates. High thermostability and an isoelectric point of 9.41 were recorded. Based on N-terminal amino acid residues, high resistance to intracellular proteases with extended in-vivo and in-vitro half-lives was predicted. TS14 is foreseen as a secretory protein with a signal peptide and an extracellular localization. Structural analysis of TS14 exhibited the dominance of helices in the secondary structure (92% coverage) with majority of residues showing high and medium solvent accessibility. High lysine content and presence of multiple nucleotide binding sites in TS14 suggests interaction with RNA/DNA and a role in their metabolism. Immunogenic profiling predicted presence of four distinct B-cell epitopes. Mutational analysis based on the single amino acid substitutions among six TS14 isolates demonstrated minor variations in structural stability; however, all the substitutions were well tolerated. Moreover, all the isolates revealed almost identical immunogenic profile with an equivocal potential to elicit the antibody-mediated immune response.

Download Full-text

Preparation and characterization of withaferin A loaded pegylated nanoliposomal formulation with high loading efficacy: In vitro and in vivo anti-tumor study

Materials Science and Engineering C ◽

10.1016/j.msec.2021.112335 ◽

2021 ◽

pp. 112335

Author(s):

Prathapan Abeesh ◽

Walsan Kalarikkal Vishnu ◽

Chandrasekharan Guruvayoorappan

Keyword(s):

High Loading ◽

Withaferin A ◽

Tumor Study

Download Full-text

Corrigendum to “Preparation and characterization of withaferin A loaded pegylated nanoliposomal formulation with high loading efficacy: In vitro and in vivo anti-tumor study” [Mater. Sci. Eng. C 128 (2021) 112335]

Materials Science and Engineering C ◽

10.1016/j.msec.2021.112432 ◽

2021 ◽

Vol 130 ◽

pp. 112432

Author(s):

Prathapan Abeesh ◽

Walsan Kalarikkal Vishnu ◽

Chandrasekharan Guruvayoorappan

Keyword(s):

High Loading ◽

Withaferin A ◽

Tumor Study

Download Full-text

In vitro-in vivo-in silico approach in biopharmaceutical characterization of ibuprofen IR and SR tablets

European Journal of Pharmaceutical Sciences ◽

10.1016/j.ejps.2015.03.027 ◽

2015 ◽

Vol 75 ◽

pp. 151-159 ◽

Cited By ~ 4

Author(s):

Sofija Beloica ◽

Sandra Cvijić ◽

Marija Bogataj ◽

Jelena Parojčić

Keyword(s):

In Silico

Download Full-text

In-vitro, in-vivo, and in-silico assessment of radical scavenging and cytotoxic activities of Oliveria decumbens essential oil and its main components

Scientific Reports ◽

10.1038/s41598-021-93535-8 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Tahereh Jamali ◽

Gholamreza Kavoosi ◽

Yousef Jamali ◽

Saeed Mortezazadeh ◽

Susan K. Ardestani

Keyword(s):

Nitric Oxide ◽

Flow Cytometry ◽

Essential Oil ◽

Cancer Cells ◽

In Silico ◽

Antioxidant Properties ◽

Radical Scavenging ◽

Main Components

AbstractWe aimed to explore and compare new insights on the pharmacological potential of Oliveria decumbence essential oil (OEO) and its main components highlighting their antioxidant activity in-vitro, in-vivo, and in-silico and also cytotoxic effects of OEO against A549 lung cancer cells. At first, based on GC–MS analysis, thymol, carvacrol, p-cymene, and γ-terpinene were introduced as basic ingredients of OEO and their in-vitro antioxidant capacity was considered by standard methods. Collectively, OEO exhibited strong antioxidant properties even more than its components. In LPS-stimulated macrophages treated with OEO, the reduction of ROS (Reactive-oxygen-species) and NO (nitric-oxide) and down-regulation of iNOS (inducible nitric-oxide-synthase) and NOX (NADPH-oxidase) mRNA expression was observed and compared with that of OEO components. According to the results, OEO, thymol, and carvacrol exhibited the highest radical scavenging potency compared to p-cymene, and γ-terpinene. In-silico Molecular-Docking and Molecular-Dynamics simulation indicated that thymol and carvacrol but no p-cymene and γ-terpinene may establish coordinative bonds in iNOS active site and thereby inhibit iNOS. However, they did not show any evidence for NOX inhibition. In the following, MTT assay showed that OEO induces cytotoxicity in A549 cancer cells despite having a limited effect on L929 normal cells. Apoptotic death and its dependence on caspase-3 activity and Bax/Bcl2 ratio in OEO-treated cells were established by fluorescence microscopy, flow cytometry, colorimetric assay, and western blot analysis. Additionally, flow cytometry studies demonstrated increased levels of ROS in OEO-treated cells. Therefore, OEO, despite showing antioxidant properties, induces apoptosis in cancer cells by increasing ROS levels. Collectively, our results provided new insight into the usage of OEO and main components, thymol, and carvacrol, into the development of novel antioxidant and anti-cancer agents.

Download Full-text

Withaferin A—A Promising Phytochemical Compound with Multiple Results in Dermatological Diseases

Molecules ◽

10.3390/molecules26092407 ◽

2021 ◽

Vol 26 (9) ◽

pp. 2407

Author(s):

Simona Bungau ◽

Cosmin Mihai Vesa ◽

Areha Abid ◽

Tapan Behl ◽

Delia Mirela Tit ◽

...

Keyword(s):

Mechanism Of Action ◽

In Silico ◽

Withania Somnifera ◽

In Vivo Studies ◽

Withaferin A ◽

Biological Targets ◽

Dermatological Diseases ◽

Anticancer Effects

Withaferin A (WFA) was identified as the most active phytocompound of the plant Withania somnifera (WS) and as having multiple therapeutic/ameliorating properties (anticancer, antiangiogenic, anti-invasive, anti-inflammatory, proapoptotic, etc.) in case of various diseases. In drug chemistry, WFA in silico approaches have identified favorite biological targets, stimulating and accelerating research to evaluate its pharmacological activity—numerous anticancer effects manifested in various organs (breast, pancreas, skin, colon, etc.), antivirals, anti-infective, etc., which are not yet sufficiently explored. This paper is a synthesis of the most relevant specialized papers in the field that are focused on the use of WFA in dermatological diseases, describing its mechanism of action while providing, at the same time, details about the results of its testing in in vitro/in vivo studies.

Download Full-text

Applying Bioinformatic Platforms, In Vitro, and In Vivo Functional Assays in the Characterization of Genetic Variants in the GH/IGF Pathway Affecting Growth and Development

Cells ◽

10.3390/cells10082063 ◽

2021 ◽

Vol 10 (8) ◽

pp. 2063

Author(s):

Sabina Domené ◽

Paula A. Scaglia ◽

Mariana L. Gutiérrez ◽

Horacio M. Domené

Keyword(s):

In Silico ◽

Growth And Development ◽

Genetic Variant ◽

Modeling Tools ◽

In Vivo Models ◽

Bioinformatic Tools ◽

Functional Assays

Heritability accounts for over 80% of adult human height, indicating that genetic variability is the main determinant of stature. The rapid technological development of Next-Generation Sequencing (NGS), particularly Whole Exome Sequencing (WES), has resulted in the characterization of several genetic conditions affecting growth and development. The greatest challenge of NGS remains the high number of candidate variants identified. In silico bioinformatic tools represent the first approach for classifying these variants. However, solving the complicated problem of variant interpretation requires the use of experimental approaches such as in vitro and, when needed, in vivo functional assays. In this review, we will discuss a rational approach to apply to the gene variants identified in children with growth and developmental defects including: (i) bioinformatic tools; (ii) in silico modeling tools; (iii) in vitro functional assays; and (iv) the development of in vivo models. While bioinformatic tools are useful for a preliminary selection of potentially pathogenic variants, in vitro—and sometimes also in vivo—functional assays are further required to unequivocally determine the pathogenicity of a novel genetic variant. This long, time-consuming, and expensive process is the only scientifically proven method to determine causality between a genetic variant and a human genetic disease.

Download Full-text