Esomeprazole and pantoprazole enhance the antiproliferative effects of cisplatin on the human neuroblastoma SH-SY5Y cell line

Background: Proton pump inhibitors (PPIs) largely used a drug to treat gastroesophageal disease such as gastric ulcers. Moreover, in recent years, several studies suggest that PPIs have an important anti-cancer effect in monotherapy and or combination with chemotherapy. The aim of this study was to investigate whether esomeprazole and pantoprazole exhibit anti-cancer effect alone or could enhance chemosensitivity on the human neuroblastoma cell line SH-SY5Y to cisplatin.Methods: The human neuroblastoma SH-SY5Y cells were cultured and treated with different concentrations of esomeprazole, pantoprazole, and cisplatin alone. Also, these cells exposed to cisplatin+ esomeprazole and cisplatin + pantoprazole combinations, respectively and incubated 24 h. The antiproliferative activities of the (PPIs) alone or in a combination of cisplatin was evaluated using the XTT colorimetric assay.Results: According to experimental data, neither PPIs showed no cytotoxicity on the human neuroblastoma cell line SH-SY5Y at all concentrations. However, when combined with cisplatin separately, they were found to have significant antiproliferative effects on the human neuroblastoma SH-SY5Y cell lines when compared to cell lines treated with cisplatin alone (p<0.05).Conclusions: Taken together, the inhibition of V-ATPase via esomeprazole and pantoprazole might enhance the chemosensitivity of cisplatin on the human neuroblastoma cell line SH-SY5Y. However, further studies are needed to be able to utilize PPIs in human neuroblastoma cells

Synthesis and Charecterisation of Methylene Blue Derivatives

Since its discovery, the fluorescent dye known as methylene blue has been employed in a variety of scientific disciplines. Methylene blue (MB) for intraoperative fluorescence imaging has only lately been employed. 10-N-carbamoyl linkage modification of Methylene blue helps us in developing compounds with off-to-on fluorescence switch in the presence of near infrared (NIR). Due to its naturally induced photodamage to normal tissues and deeper penetration of lesion regions when compared to UV and visible light, near infrared (NIR) light is considered favorable among the broad radiation range. The previously synthesized derivatives are not completely soluble in water, they require other co-solvents, acetone etc. So in this study branched polyethyleneimine which is a water soluble polymer is used to conjugate with methylene blue to effect the 10-N-Carbamoyl linkage. The branched structure of polyethyleneimine has several primary amines, so the amount of methylene blue that can be conjugated can also be varied. The carbamoyl linkage with methylene blue is also sensitive to hydroxyl radical. Using this hypothesis the activity was verified using hemoproteins such as hemoglobin. Subsequently this property can be used in the lysis of the malaria parasite. MB and PEI conjugate was synthesized and tested in vitro for the cleavage of the 10-N-Carbamoyl linkage. As the result, PEI-conjugated MB successfully demonstrated the cleavage and also sensitivity to ROS-induce cleavage to release intact MB

pH-Responsive Poly(Ethylene Glycol)- block -Polylactide Micelles for Tumor-Targeted Drug Delivery

This document is the Accepted Manuscript version of a Published Work that appeared in final form in Biomacromolecules, copyright © American Chemical Society after peer review and technical editing by publisher. To access the final edited and published work see: http://dx.doi.org/10.1021/acs.biomac.7b00509A biodegradable micellar drug delivery system with a pH-responsive sheddable PEG shell was developed using an acetal-linked poly(ethylene glycol)-block-polylactide (PEG-a-PLA) copolymer and applied to the tumoral release of paclitaxel (PTX). The micelles with a diameter of ∼100 nm were stable in PBS at pH 7.4, started shedding the shell and aggregating slowly at pH 6.5, and decomposed faster at pH 5.5. PTX-loaded micelles (M-PTX) with a drug loading of 6.9 wt % exhibited pH-triggered PTX release in simulated tumoral acidic environments corresponding to the extracellular and intracellular spaces. In vitro experiments showed that the micelles were noncytotoxic to different cell lines, while M-PTX inhibited the proliferation and promoted the apoptosis of Hela cells. An in vivo study with Hela tumor-bearing mice indicated that M-PTX efficiently inhibited tumor growth. Because of these properties, the PEG-a-PLA micellar system appears to have bright prospects as a tumor-targeting drug carrier.China Postdoctoral Science Foundation || General Program, No. 18 2015M580640 National Natural Science Foundation of China || General Program, No.19 2157405

Nanotransportadores híbridos para el tratamiento selectivo de enfermedades complejas

Tesis de la Universidad Complutense de Madrid, Facultad de Farmacia, leída el 19-07-2018Esta tesis doctoral se centra en el diseño, síntesis y evaluación de nanosistemas inteligentes para el tratamiento de enfermedades complejas como el cáncer y la fibrosis. El cáncer es una enfermedad maligna y multifacética con una incidencia significativa en la población mundial. Tradicionalmente se ha tratado con cirugía, radioterapia y quimioterapia. Sin embargo, la extirpación total de tumores mediante cirugía no siempre es posible y la radioterapia y la quimioterapia no presentan selectividad hacia las masas tumorales. Por lo tanto, estos tratamientos no suelen ser eficaces para la eliminación completa de los tumores y su eficacia se ve a menudo limitada por la aparición de efectos secundarios graves. La nanomedicina es un subcampo emergente de la nanotecnología que trata de proporcionar un tratamiento más eficaz de enfermedades complejas. El uso de nanomateriales en el cáncer se basa en su capacidad de acumularse en masas tumorales debido a las características particulares de estos tejidos malignos. Este efecto recibe el nombre de Enhance Permeation and Retention (EPR) effect, y su descubrimiento en 1986 supuso el punto de partida para la nanomedicina aplicada a la terapia oncológica. De esta manera, el uso de nanopartículas como portadores de fármacos permite transportar una alta cantidad de fármacos antitumorales de forma selectiva y controlada al tumor,reduciendo la aparición de los efectos secundarios asociados a la alta toxicidad de estos fármacos...This doctoral thesis is focused on the design, synthesis and evaluation of smart nanosystems for the treatment of complex diseases such as cancer and fibrosis. Cancer is a malignant and multifaceted disease with a significant incidence in the world population. Surgery, radiotherapy and chemotherapy have been traditionally employedfor cancer treatment. However, the complete eradication of tumors simply by surgical operations is not always possible, while radiotherapy and chemotherapy affects equally to both, healthy and tumoral cells. Therefore, these treatments are not usually effective for the definitive elimination of tumors being their effectiveness limited by the appearance of severe side effects, because of their lack of selectivity. Nanomedicine is an emergent and potent sub-field of nanotechnology which tries to provide more efficient treatment for complex diseases. The use of nanomaterials in cancer is based on their capacity to be accumulated in tumoral masses due to the particular characteristics of malignant tissues. This effect is the so called Enhance Permeation and Retention (EPR) effect, and its discovery in 1986 supposed the starting point for the nanomedicine applied to oncological therapy...Fac. de FarmaciaTRUEunpu

Annexin A5–methotrexate: a novel targeted therapeutic against breast cancer and leukemia

One of the most challenging issues of chemotherapy in cancer treatments relies on specifically targeting and carrying the drug to the tumor cells. While tumor ablation surgery can be efficient, metastatic cancers are more challenging to treat because of the spread distribution of tumor cells in the body. Methotrexate is an FDA approved antimetabolite drug for the treatment of breast cancer and acute lymphoblastic leukemia. However, despite several conjugation attempts to optimize its delivery to cancer cells, methotrexate still does not offer a reliable enough therapeutic effect in the clinic. We have investigated the conjugation of methotrexate to annexin A5, a human protein that specifically binds to the cancer cell marker phosphatidylserine. Once conjugated to annexin A5, methotrexate’s cytostatic effect would be specifically delivered to cancer cells without causing off-target damage to healthy cells. This novel drug conjugate has been designed as a response to the currently urgent clinical needs for a targeted and specific chemotherapeutic treatment for triple negative breast cancer and lymphoblastic leukemia. The successful conjugation of methotrexate to annexin A5 was confirmed by measuring the final concentration of annexin A5 and methotrexate in the conjugate. The increase in size of the conjugate was confirmed by SDS-PAGE. Cell viability assays of free and conjugated methotrexate were conducted on both tumor and non-tumor cells. The conjugate induced a toxicity that was statistically equivalent to the one delivered by the free drug on the triple negative breast cancer 4T1 and EMT6 cell lines, and on the lymphoblastic leukemia P388 cell line. However, while free methotrexate induces a cell mortality in healthy endothelial HUVEC and HAAE-1 cells, the conjugate does not cause any significant decrease of the cell viability, confirming its specificity towards tumor cells

Smart and Functional Polymers

This book is based on the Special Issue of the journal Molecules on “Smart and Functional Polymers”. The collected research and review articles focus on the synthesis and characterization of advanced functional polymers, polymers with specific structures and performances, current improvements in advanced polymer-based materials for various applications, and the opportunities and challenges in the future. The topics cover the emerging synthesis and characterization technology of smart polymers, core?shell structure polymers, stimuli-responsive polymers, anhydrous electrorheological materials fabricated from conducting polymers, reversible polymerization systems, and biomedical polymers for drug delivery and disease theranostics. In summary, this book provides a comprehensive overview of the latest synthesis approaches, representative structures and performances, and various applications of smart and functional polymers. It will serve as a useful reference for all researchers and readers interested in polymer sciences and technologies

Carbon nanotube and graphene cellular impact towards their possible use as nanovectors for anticancer therapy and cell targeting

In the last two decades, lots of progresses have been made in the application of nanomaterials in biology, giving rise to a new revolutionary field of the biomedical research that is called nanomedicine. Carbon nanotubes (CNTs) and graphene are among the most attractive candidates under investigation in this context. Such carbon-based nanomaterials possess outstanding chemico-physical properties and their high surface area provides multiple attachment sites for almost all the molecules of biological interest. In fact, chemical strategies allow for their conjugation with DNA, proteins, peptides and small drugs. The so-functionalized materials have shown great promises in several biomedical contexts, from diagnosis to tissue regeneration. In addition, the ability of CNTs and graphene to enter and accumulate inside the cells makes them good candidates as nanovectors for drugs. In vitro and in vivo studies showed how these nanomaterials could enhance the drug accumulation inside the cells and its bioavailability. CNTs and graphene are also able to passively target tumor tissues exploiting the so-called enhanced permeability and retention effect, and actively, following the conjugation with targeting moieties. On the other hand, the use of these materials in nanomedicine will be approved only after the demonstration of their safety in terms of tissue damage, carcinogenicity and proinflammatory response. Different approaches have been used to make these materials biocompatible and recently, it has been demonstrated that functionalized CNTs and graphene oxide can be degraded by oxidative enzymes. However, the results obtained on CNT and graphene toxicity are often conflicting; thus, further investigations are needed. In the first introductive chapter of this thesis, an overview of the general features of nanomaterials is given. The focus goes then on CNTs and graphene. First, we will see how their biocompatibility, biodegradability and in vivo fate strictly depend on several factors. The mechanisms by which the two nanomaterials are able to enter the cells are also illustrated. In the second part of the introduction, the potential of CNTs and graphene in targeted drug delivery is described, focusing on anticancer therapy. The three following chapters illustrate the results obtained by three related studies carried out during my PhD internship. Results described in the second chapter shed more light on the impact of CNTs on living cells. CNTs having single walls (SWCNTs) were dispersed with the biocompatible protein bovine serum albumin (BSA). BSA protein showed to be a good dispersant agent for the CNTs and was able to enhance their biocompatibility. The impact of the protein-coated materials on cell vital parameters, such as viability, activation and interaction/internalization mechanisms, is described. In addition the effect of nanotubes on the plasma membrane dielectric characteristics and ion flux, very poorly 2 Abstract investigated up to date, is presented. In the following chapter, a cisplatin prodrug was encapsulated within the inner cavity of two types of multi-walled CNTs (MWCNTs) having different diameters, in order to allow a controlled release inside the cells. The efficacy of the complexes was investigated on human cervix cancer cells and compared to murine macrophages. The latter were also used to evaluate the possibility of a proinflammatory effect. Results on cell viability, cell activation and cell uptake show that CNTs are promising nanocarriers to improve the accumulation of a chemotherapeutic drug inside the cells, without inducing a high proinflammatory response. In addition, by tuning CNT diameter it is possible to control the time of release of the drug prolonging its anticancer efficacy. The fourth chapter gives more insights about the impact of different types of graphene on cells, focusing on macrophages. Results evidenced a specific cytotoxic effect of graphene oxide (GO) towards this cell population among the other murine immune cells. The mechanisms by which macrophages internalize GO are also elucidated. In addition, GO was conjugated with lysozyme protein and was tested for its ability to selectively target a B cell model overexpressing a lysozyme-specific B cell receptor. Results showed that graphene is able to target B cells in a selective manner, thus suggesting its possible application in therapies where the specific elimination of B lymphocytes is required. Finally, a conclusive chapter summarizes the main findings obtained during my doctoral work, focusing, in particular, on future perspectives

Stealth properties of poly(ethylene oxide)-based triblock copolymer micelles: A prerequisite for a pH-triggered targeting system

Evaluation of the biocompatibility of pH-triggered targeting micelles was performed with the goal of studying the effect of a poly(ethylene oxide) (PEO) coating on micelle stealth properties. Upon protonation under acidic conditions, pH-sensitive poly(2-vinylpyridine) (P2VP) blocks were stretched, exhibiting positive charges at the periphery of the micelles as well as being a model targeting unit. The polymer micelles were based on two different macromolecular architectures, an ABC miktoarm star terpolymer and an ABC linear triblock copolymer, which combined three different polymer blocks, i.e. hydrophobic poly(E-caprolactone), PEO and P2VP. Neutral polymer micelles were formed at physiological pH. These systems were tested for their ability to avoid macrophage uptake, their complement activation and their pharmacological behavior after systemic injection in mice, as a function of their conformation (neutral or protonated). After protonation, complement activation and macrophage uptake were up to twofold higher than for neutral systems. By contrast, when P2VP blocks and the targeting unit were buried by the PEO shell at physiological pH, micelle stealth properties were improved, allowing their future systemic injection with an expected long circulation in blood. Smart systems responsive to pH were thus developed which therefore hold great promise for targeted drug delivery to an acidic tumoral environment