Hyaluronan-conjugated liposomes encapsulating gemcitabine for breast cancer stem cells (original) (raw)
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Biochimica et Biophysica Acta (BBA) - Biomembranes, 2013
Pancreatic adenocarcinoma is often diagnosed when metastatic events have occurred. The early spread of circulating cancer cells expressing the CD44 receptor may play a crucial role in this process. In this study, we have investigated the cellular delivery ability and both in vitro and in vivo anti-tumoral activity of liposomes conjugated with two different low molecular weight hyaluronic acids (HA 4.8 kDa and HA 12 kDa), the primary ligand of CD44, and containing a lipophilic gemcitabine (GEM) pro-drug. By confocal microscopy and flow cytometry analyses, we demonstrate that the cellular uptake into a highly CD44-expressing pancreatic adenocarcinoma cell line is higher with HA-conjugated (12 kDa > 4.8 kDa) than non-conjugated liposomes. Consistently, in vitro cytotoxic assays display an increased sensitivity towards GEM containing HA-liposomes, compared to non-conjugated liposomes. Conversely, CD44 non-expressing normal cells show a similar uptake and in vitro cytotoxicity with both HA-conjugated and non-conjugated liposomes. Furthermore, we demonstrate that the HA-liposomes are taken up into the cells via lipid raft-mediated endocytosis. All the liposome formulations containing GEM show a higher antitumoral activity than free GEM in a mouse xenograft tumor model of human pancreatic adenocarcinoma. The 12 kDa HA-liposomes have the strongest efficiency, while non-conjugated liposomes and the 4.8 kDa HA-liposomes are similarly active. Taken together, our results provide a strong rationale for further development of HA-conjugated liposomes to treat pancreatic adenocarcinoma.
Hyaluronic acid-coated liposomes for active targeting of gemcitabine
European Journal of Pharmaceutics and Biopharmaceutics, 2013
The aim of this work was the preparation, characterization, and preliminary evaluation of the targeting ability toward pancreatic adenocarcinoma cells of liposomes containing the gemcitabine lipophilic prodrug [4-(N)-lauroyl-gemcitabine, C12GEM]. Hyaluronic acid (HA) was selected as targeting agent since it is biodegradable, biocompatible, and can be chemically modified and its cell surface receptor CD44 is overexpressed on various tumors.
Journal of drug targeting, 2018
Multi Drug Resistance (MDR) of cancer cells is a constant threat to the clinically used drugs as well as new drug development. In present, work we aimed to assess in-vitro as well as in-vivo efficacy of the developed Imatinib loaded liposomes in MDR cancer. An array of tests was also carried out to comprehensively understand the bio-mechanism that enable these nanocarriers to modulate P-gp activity. Hyaluronic acid coated, Imatinib mesylate containing lipsomes (HA-LIPO-IM) were analysed through in-vitro and in-vivo studies in MDR cancer cells and tumour models. Effect of developed hyaluronated liposomes on various biomolecular mechanisms was also evaluated. Around 3.5 times lower IC50 for HA-LIPO-IM in comparison to drug solution in HT-29 and Colo-320 cells proved the enhanced action of the drug in MDR cells. HA-LIPO formulation were demonstrated to have inhibitory effect on ATPase enzyme. Molecular masking of Imatinib mesylate and CD-44 mediated endocytosis were also found responsi...
Expert Opinion on Drug Delivery, 2020
Background: Phytosterols significantly reduce the risk of cancer by directly inhibiting tumor growth, inducing apoptosis, and inhibiting tumor metastasis. Stigmasterol (STS), a phytosterol, exhibits anticancer effects against various cancers, including breast cancer. Chemotherapeutics, including doxorubicin (DOX), might act synergistically with phytosterol against the proliferation and metastasis of breast cancer. Although such compounds can show potential anticancer activity, their combined effect with suitable formulation has not investigated yet. Methods: Hyaluronic acid (HA)-modified PEGylated DOX-STS loaded phyto-liposome was fabricated via a thin-film hydration method. The prepared phyto-liposome was optimized with regards to its physicochemical and other properties. Further, in vitro and in vivo study was carried out in breast cancer cells expressing a different level of CD44 receptors. Results: The particle size of prepared HA-DOX-STS-lipo was 173.9 ± 2.4 nm, and showed pH-depended DOX release, favoring the effective tumor targetability. The in vitro anticancer activity of HA-DOX-STS-lipo was significantly enhanced in MDA-MB-231, CD44-overexpressing cells relative to MCF-7 cells demonstrating HA-mediated targeting effect. HA-DOX-STS-lipo accumulated more and increased antitumor efficacy in the MDA-MB-231 xenograft tumor model expressing high levels of CD44, suggesting the potential of carrier system towards CD44-overexpressing tumors.
Current Drug Delivery, 2007
The effects of lipid composition and preparation conditions on the physicochemical and technological properties of gemcitabine-loaded liposomes, as well as the in vitro anti-tumoral activity of various liposome formulations were investigated. Three liposome formulations were investigated: DPPC/Chol/Oleic acid (8:3:1 molar ratio, liposomes A), DPPC/Chol/DPPS (6:3:1 molar ratio, liposomes B) and DPPC/Chol/DSPE-MPEG (6:3:1 molar ratio, liposomes C). Multilamellar liposomes were prepared by using the TLE, FAT and DRV methods, while small unilamellar liposomes were obtained by extrusion through polycarbonate filters. Light scattering techniques were used to characterize liposome formulations. Loading capacity and release profiles of gemcitabine from various liposome formulations were also investigated. Caco-2 cells were used to evaluate in vitro the antitumoral activity of gemcitabine-loaded liposomes with respect to the free drug and also the intracellular drug uptake. Preparation methods and liposome lipid composition influenced both physicochemical parameters and drug delivery features. Liposomes with a size ranging from 200 nm to 7 m were obtained. The gemcitabine entrapment was higher than that expected probably due to an interaction with the liposome lipid components. The following decreasing loading capacity order was observed: liposome B>liposome C>liposome A. Gemcitabine release from various liposome formulations is modulated by two different processes, i.e. desorption from and permeation through liposomal bilayers. MTT assay showed a greater cytotoxic effect of gemcitabine-loaded liposomes with respect to the free drug. The following decreasing anticancer activity order was observed between the various liposome formulations: liposome C>liposome A>liposome B. The increased anticancer activity is correlated to the ability of the colloidal carrier to increase the intracellular drug uptake. Due to the encouraging results and to the high liposome modularity various applications of potential therapeutic relevance can be envisaged for liposomes.
Gemcitabine-loaded liposomes: rationale, potentialities and future perspectives
International Journal of Nanomedicine, 2012
This review describes the strategies used in recent years to improve the biopharmaceutical properties of gemcitabine, a nucleoside analog deoxycytidine antimetabolite characterized by activity against many kinds of tumors, by means of liposomal devices. The main limitation of using this active compound is the rapid inactivation of deoxycytidine deaminase following administration in vivo. Consequently, different strategies based on its encapsulation/complexation in innovative vesicular colloidal carriers have been investigated, with interesting results in terms of increased pharmacological activity, plasma half-life, and tumor localization, in addition to decreased side effects. This review focuses on the specific approaches used, based on the encapsulation of gemcitabine in liposomes, with particular attention to the results obtained during the last 5 years. These approaches represent a valid starting point in the attempt to obtain a novel, commercializable drug formulation as already achieved for liposomal doxorubicin (Doxil ® , Caelyx ®).
Lipid-Based Nanovectors for Targeting of CD44-Overexpressing Tumor Cells
Journal of drug delivery, 2013
Hyaluronic acid (HA) is a naturally occurring glycosaminoglycan that exists in living systems, and it is a major component of the extracellular matrix. The hyaluronic acid receptor CD44 is found at low levels on the surface of epithelial, haematopoietic, and neuronal cells and is overexpressed in many cancer cells particularly in tumour initiating cells. HA has been therefore used as ligand attached to HA-lipid-based nanovectors for the active targeting of small or large active molecules for the treatment of cancer. This paper describes the different approaches employed for the preparation, characterization, and evaluation of these potent delivery systems.
Pharmaceutical Development and Technology, 2017
The aim of this study is to formulate and compare the physicochemical properties of negatively-charged liposomes and poly-lactide-co-glycolide (PLGA) nanoparticles loaded with gemcitabine hydrochloride. The influence of the formulation variables on the liposome and nanoparticle properties on particle size, zeta potential, encapsulation efficiency and drug release were evaluated. Although the PEGylated nanoparticles and PEGylated liposomes were of the same size (~200 nm), the encapsulation efficiency was 1.4 times higher for PEGylated liposomes than for PEGylated nanoparticles. The optimized formulation of PEGylated liposomes and PEGylated nanoparticles had 26.1 ± 0.18 and 18.8 ± 1.52% encapsulation efficiency, respectively. The release of drug from the PEGylated liposomes and PEGylated nanoparticles exhibited a biphasic pattern that was characterized by a fast initial release during the first 2 h followed by a slower continuous release. Transmission electron microscopy (TEM) images identified separate circular structures of the liposomes and nanoparticles. The in vitro cytotoxicity of the optimized formulations was assessed in MCF-7 and MDA-MB-231 cells, the results showed that the cytotoxicity effect of the gemcitabine hydrochloride loaded liposomes and nanoparticles were more than commercial product Gemko ® and gemcitabine hydrochloride solution.
Enhanced Tumor Targeting and Antitumor Activity of Gemcitabine Encapsulated Stealth Liposome’s
Indian Journal of Pharmaceutical Education and Research, 2015
Introduction Cancer is a term used for diseases in which abnormal cells divide without control and are able to invade other tissues. Cancer cells can spread to other parts of the body through the blood and lymph systems. Cancer is not just one disease but many diseases. There are more than 200 different types of cancer [1-4]. For instance, although there are numerous anticancer agents that are highly cytotoxic to tumor cells in vitro, the lack of selective antitumor effect in vivo precludes their use in clinic. One of the major limitations of antineoplastic drugs is their low therapeutic index (TI), i.e. the dose required to produce anti-tumor effect is toxic to normal tissues. Liposomes are spherical vesicles composed of lipid bilayers arranged around a central aqueous core. The particle size of liposomes ranges from 20 nm to 10 μm in diameter. They can be composed of natural constituents such as phospholipids and may mimic naturally occurring cell membranes. Liposomes have the ability to incorporate lipophilic and hydrophilic drugs within their phospholipid membrane or they can encapsulate hydrophilic compounds within the aqueous core [6]. Gemcitabine is new cytotoxic drug but some of limitations while its use likes it suppress the activity of bone marrow i.e. effect on blood forming cells. Higher water solubility needs to improve encapsulation efficiency for better therapeutics effect. Stealth liposomes by pH gradient technology lower half life-7-18 min, unable to deliver by oral and other route. Higher dose-1000-1250 mg/m2 require against malignancies are effective against various solid tumor like colon, lungs, breast etc [7, 8]. Sterically stabilized liposomes can be formulated by incorporating hydrophilic long-chain polymers (PEG) in the bilayer which can form a coat on the liposome surface and repel opsonin penetration and adsorption. Reduction in 'marking' by opsonins leads to slower uptake of these liposomes (LCL) by the cells of reticuloendothelial system (RES) [9]. In present investigation focuses on to perform innovative research work is to avoid the problem associated with gemcitabine use and effective against solid tumor with minimum toxic effect by incorporating it in stealth liposomes. Materials and Methods Materials Gemcitabine was obtained as gift sample from Sun Pharma Pvt Ltd, Vadodara, (DPPC) 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine, Soya PC, (DSPE-MPEG-2000) 1,2-Distearoyl-sn-glycero-3phosphoethanolamine-methyl-polyethyleneglycol conjugate-2000 Na+ salt, Cholesterol was obtained from Lipoid GmbH, Ludwigshafen, Germany, Chloroform, Methanol, and other chemical was purchased from Loba Chemicals, Mumbai. All other solvent and reagents were of analytical grade.
Journal of Nanoscience and Nanotechnology, 2008
Anaplastic thyroid carcinoma is one of the most aggressive and lethal solid carcinomas affecting humans. A major limit of the chemotherapeutic agents is represented by their low therapeutic index. In this work, we investigated the possibility of improving the anti-tumoral activity of gemcitabine by using pegylated unilamellar liposomes. Liposomes were made up of 1,2-dipalmitoyl-sn-glycero-3-phospocholine monohydrate/cholesterol/N-(carbonyl-methoxypolyethylene glycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (6:3:1 molar ratio) and they were prepared with a pH gradient to improve the gemcitabine loading capacity. The anti-tumoral efficacy of the liposomal formulation was tested in vitro on human anaplastic thyroid carcinoma cells (ARO) in culture, comparing the effects with those of the free drug. Gemcitabine-loaded unilamellar liposomes had a mean size ∼200 nm with a zeta potential ∼–2mV. The liposomal carrier noticeably improved the anti-tumoral activity of gemcitabine ...