Ph-responsive liposomal systems for site-specific pulmonary delivery of anti-tubercular drugs

Tuberculosis (TB) is an infectious disease that has been reported to be the ninth leading cause of death worldwide, even though mostly considered as a poverty related disease. Despite the existence of potent anti-tubercular drugs (ATBDs), such as rifampicin (RIF) and isoniazid (INH), TB remains the major killer among many microbial diseases over the last five years. Although several factors are to be blamed for this deadly status, the most crucial issues encompass both the self-defensiveness of the causative agent (Mycobacterium tuberculosis), including its intra-macrophage location that compromises ATBDs accessibility, and the widespread/off target distribution of ATBDs. The need for novel drug delivery strategies therefore arises to provide selective distribution of ATBDs at the infected site. Among the drug vehicles explored in this field, liposomes have been reported to be the most suitable drug carriers due to their rapid uptake by alveolar macrophages, where M. tuberculosis often resides. Since liposomes experience media of different pH throughout the cell uptake process (endocytosis/phagocytosis), the use of pH change as a stimulus for controlled release looks promising for enhancing intra-macrophage delivery and minimizing premature ‘off-target’ release of ATBDs. However, the costly status of liposome technology, due to the use of sophisticated procedures and expensive materials (especially for pH-dependent delivery, where special lipids are required), may preclude wider developments of liposomal products, especially for the developing world. This study aimed at investigating liposomal encapsulation of pH-sensitive and fluorescent hydrazone derivatives of INH using crude soybean lecithin, as a cost-effective option for site-specific delivery combined with potential bio-imaging features. Another objective was to explore encapsulation of INH hydrazone derivatives with and without RIF in liposomes using a simple and organic solvent-free preparation method. Initially, INH was coupled with 4-hydroxy-benzaldehyde to yield a conjugate (INH-HB) that was encapsulated in liposomes using film hydration method with acceptable encapsulation efficiency (î), about 89 %. The prepared INH-HB loaded liposomes (IHL) were characterized by means of dynamic light scattering (DLS), transmission electron microscopy (TEM), differential scanning calorimetry (DSC) and X-ray diffraction (XRD). The release of INH from IHL was evaluated over 12 hours in media of different pH using dialysis. As hypothesized, pH dependent release of INH from IHL was observed with 22, 69, 83 and 100 % release obtained in media of pH 7.4, 6.4, 5.4 and 4.4, respectively. From this experimental trial, further development was undertaken by conjugating INH to a hydrophobic fluorescent tag, zinc (II) phthalocyanine (PC), through hydrazone linkages. The obtained conjugate (PC-INH) was loaded into liposomes (PIL) that were characterized using various spectroscopic techniques, including UV-Vis absorption and energy dispersive X-ray spectroscopy, which suggested the presence of PC-INH within the lipid bilayers. The release study performed in different pH media revealed 22, 41, 97 and 100 % of INH, respectively released at pH 7.4, 6.4, 5.4 and 4.4. This confirmed the potential of pH-triggered drug release from liposomes loaded with hydrazone drug derivatives. In addition, successful encapsulation of PC-INH using crude soybean lecithin inspired a new opening towards development of multimodal liposomes that could achieve controlled drug release with added benefits of image-guided biological tracking. However, the hydrophobic nature of PC-INH requires an effective strategy that could improve its solubility and favour extensive development. In this context, the tetra-substituted structure of PC-INH brought up the hypothesis that cyclodextrin (CD) complexation would facilitate PC-INH encapsulation in liposomes using an organic solvent-free method, called here the “heating method” (HM). Inclusion complexes of PC-INH with various CDs were therefore investigated, with gamma-CD complex (CP) giving the best results. These complexes were prepared in both solution and solid-state and further comprehensively characterized using UV-Vis spectroscopy, magnetic circular dichroism, NMR spectroscopy, diffusion ordered spectroscopy, DSC, XRD and Fourier transform infrared spectroscopy. CP-loaded liposomes prepared using HM exhibited greater î than film hydration liposomes, about 70 % versus 56 %, respectively. The HM-liposomal system (CPL) exhibited potentially useful nano particulate characteristics (i.e. mean particle size 240 nm and Zeta potential –57 mV), which remained unchanged over 5 weeks of stability study at 4 °C, and pH-dependent INH release behaviour alike PIL. Furthermore, CP was co-encapsulated with rifampicin (RIF) in liposomes using HM to investigate the possibility for future combination therapy. 1H-NMR spectroscopy, DSC, XRD and photophysical studies were performed for molecular assessment of the cargo in CP-RIF co-loaded liposomes (CPRL). The mean particle size, Zeta potential and î of CPRL were respectively 594 nm, –50 mV, 58 % for CP and 86 % for RIF. CPRL exhibited much higher release rates for both INH and RIF at pH 6.4, compared to those tested at pH 7.4. In addition, there was no cytotoxicity on HeLa cells, but attractive lung fibroblasts and epithelial cells uptake and viability. Hence, CPRL are promising for targeted ATBD delivery to alveolar macrophages following pulmonary administration. Overall, the developed pH-responsive liposomal system holds the promise for new openings towards wider developments of multifunctional liposomes for site-specific controlled pulmonary delivery of antimicrobials drugs

Inhalable particulate systems for anti-tubercular drug delivery

Tuberculosis (TB) is a deadly infectious microbial disease that is currently dominating public health concerns. Among the pharmacological issues in the management of TB are the poor bioavailability of some anti-TB drugs, mostly due to the fast first-pass metabolism, and high drug load needed for combination therapy. These result in a lengthy treatment with several adverse effects causing decreased patient compliance. These factors often lead to the therapeutic failure and promote the development of drug resistant strains, justifying the urgent need for new therapeutic strategies. Liposomes are lipid-based particulate vehicles known to be the most clinically appointed drug carriers currently. Liposomal systems are reported to be rapidly engulfed by macrophages - where the mycobacterium often resides. This makes liposomes appropriate vehicles for targeted anti-TB drug delivery. Many research groups have reported the potential of liposomes systems to deliver anti-TB drugs. However, the costly formulation status of liposomes, due the use of expensive synthetic or highly purified natural phospholipids, is a limitation to the treatment of a poverty related infectious disease like TB. The aim of this study was to design and develop liposomes for pulmonary delivery of anti-TB drugs using crude soybean lecithin (CL) and its purirified version. CL is an FDA- approved naturally occurring phospholipid mixture that is quite cheap and readily available. Various liposome batches were prepared using a film hydration method and characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM). Liposomes composed of CL and cholesterol (Chol) in a 3:1 mass ratio were selected for drug encapsulation based on the following characteristics: polydispersity index (PDI, 0.28), mean particles sizes (PS, 502 nm) and zeta potential (ZP, -56 mV). Isoniazid (INH) was encapsulated as a model drug using a freeze-thaw loading technique and an HPLC method was validated for quantitative analysis. The physicochemical properties of INH-loaded liposomes were comprehensively investigated using thermal, microscopy and spectroscopic techniques. This formulation showed a high encapsulation efficiency (%EE) of 78%, much better than the liposomes made from purified lecithin, 20%. Other characteristics of INH- loaded liposomes, which make them attractive for pulmonary TB therapy, are presented in this dissertation. These include a controlled release of 50% of the encapsulated INH over 12 hours. Finally, rifampicin (RIF) was added as a hydrophobic model drug and several evaluations were conducted on these dual drug-loaded liposomes. Of particular interest, it was noted that the dual drug-loaded liposomes made of CL alone showed the highest %EE (59% for INH and 90% for RIF) compared to those containing Chol or those made of purified lecithin. Surprisingly, the average PS of the dual CL-based liposomes (1114 nm) was in the size range reported for optimum deep lung deposition and macrophage uptake. In addition, the mean ZP of these liposomes (-63 mV) seems to be favourable for their shelf stability and internalization by macrophages. Overall, these findings show that the dual CL-based liposomes developed would be promising for macrophage-targeting pulmonary delivery of anti-TB drugs

General Perception of Liposomes: Formation, Manufacturing and Applications

Liposomes are currently part of the most reputed carriers for various molecular species, from small and simple to large and complex molecules. Since their discovery, liposomes have been subject to extensive evolution, in terms of composition, manufacturing and applications, which led to several openings in both basic and applied life sciences. However, most of the advances in liposome research have been more devoted to launching new developments than improving the existing technology for potential implementation. For instance, the evolution of the conventional lipid hydration methods to novel microfluidic technologies has permitted upscale production, but with increase in manufacturing cost and persistent use of organic solvents. This chapter intends to present general concepts in liposome technology, highlighting some longstanding bottlenecks that remain challenging to the preparation, characterization and applications of liposomal systems. This would enhance the understanding of the gaps in the field and, hence, provide directions for future research and developments

Preparation and characterization of isoniazid-loaded crude soybean lecithin liposomes

Unexpected substituent-dependent regioselectivty challenges in the synthesis of C-benzylated (N-arylcarbamoyl)phosphonate esters have been resolved. The C-benzylated N-furfurylcarbamoyl derivative showed low micromolar PfLDH inhibition, while one of the C-benzylated N-arylcarbamoyl analogues was active against Nagana Trypanosoma brucei parasites which are responsible for African trypanosomiasis in cattle

The pharmacology of plant virus nanoparticles

The application of nanoparticles for medical purposes has made enormous strides in providing new solutions to health problems. The observation that plant virus-based nanoparticles (VNPs) can be repurposed and engineered as smart bio-vehicles for targeted drug delivery and imaging has launched extensive research for improving the therapeutic and diagnostic management of various diseases. There is evidence that VNPs are promising high value nanocarriers with potential for translational development. This is mainly due to their unique features, encompassing structural uniformity, ease of manufacture and functionalization by means of expression, chemical biology and self-assembly. While the development pipeline is moving rapidly, with many reports focusing on engineering and manufacturing aspects to tailor the properties and efficacy of VNPs, fewer studies have focused on gaining insights into the nanotoxicity of this novel platform nanotechnology. Herein, we discuss the pharmacology of VNPs as a function of formulation and route of administration. VNPs are reviewed in the context of their application as therapeutic adjuvants or nanocarrier excipients to initiate, enhance, attenuate or impede the formulation's toxicity. The summary of the data however also underlines the need for meticulous VNP structure-nanotoxicity studies to improve our understanding of their in vivo fates and pharmacological profiles to pave the way for translation of VNP-based formulations into the clinical setting

Synthesis of pH sensitive dual capped CdTe QDs: their optical properties and structural morphology

We herein report five different types of thiol dual capped cadmium tellurite quantum dots (CdTe QDs) namely glutathionemercapto-propanoic acid (QD 1), glutathione-thiolglycolic acid (QD 2), L-cysteine-mercapto-propanoic acid (QD 3), L-cysteinethiol-glycolic acid (QD 4) and mercapto-propanoic acid-thiol-glycolic (QD 5). Dual-capped CdTe QDs were prepared using a one pot synthetic method. Cadmium acetate and sodium tellurite were respectively used as cadmium and tellurium precursors. Photo-physical properties of the synthesized QDs were examined using UV-Vis and photoluminescence spectroscopy while structural characterization was performed by means of transmission electron microscopy (TEM) and Fourier transform infrared (FTIR) spectroscopy

The in vivo fate of tobacco mosaic virus nanoparticle theranostic agents modified by the addition of a polydopamine coat

Plant virus nanoparticles (VNPs) have multiple advantages over their synthetic counterparts including the cost-effective large-scale manufacturing of uniform particles that are easy to functionalize. Tobacco mosaic virus (TMV) is one of the most promising VNP scaffolds, reflecting its high aspect ratio and ability to carry and/or display multivalent therapeutic ligands and contrast agents. Here we investigated the circulation, protein corona, immunogenicity, and organ distribution/clearance of TMV particles internally co-labeled with cyanine 5 (Cy5) and chelated gadolinium (Gd) for dual tracking by fluorescence imaging and optical emission spectrometry, with or without an external coating of polydopamine (PDA) to confer photothermal and photoacoustic capabilities. The PDA-coated particles (Gd-Cy5-TMV-PDA) showed a shorter plasma circulation time and broader distribution to organs of the reticuloendothelial system (liver, lungs, and spleen) than uncoated Gd-Cy5-TMV particles (liver and spleen only). The Gd-Cy5-TMV-PDA particles were surrounded by 2-10-fold greater protein corona (containing mainly immunoglobulins) compared to Gd-Cy5-TMV particles. However, the enzyme-linked immunosorbent assay (ELISA) revealed that PDA-coated particles bind 2-fold lesser to anti-TMV antibodies elicited by particle injection than uncoated particles, suggesting that the PDA coat enables evasion from systemic antibody surveillance. Gd-Cy5-TMV-PDA particles were cleared from organs after 8 days compared to 5 days for the uncoated particles. The slower tissue clearance of the coated particles makes them ideal for theranostic applications by facilitating sustained local delivery in addition to multimodal imaging and photothermal capabilities. We have demonstrated the potential of PDA-coated proteinaceous nanoparticles for multiple biomedical applications

Clinically established biodegradable long acting injectables: An industry perspective

Long acting injectable formulations have been developed to sustain the action of drugs in the body over desired periods of time. These delivery platforms have been utilized for both systemic and local drug delivery applications. This review gives an overview of long acting injectable systems that are currently in clinical use. These products are categorized in three different groups: biodegradable polymeric systems, including microparticles and implants; micro and nanocrystal suspensions and oil-based formulations. Furthermore, the applications of these drug delivery platforms for the management of various chronic diseases are summarized. Finally, this review addresses industrial challenges regarding the development of long acting injectable formulations

Clinically established biodegradable long acting injectables: An industry perspective

Long acting injectable formulations have been developed to sustain the action of drugs in the body over desired periods of time. These delivery platforms have been utilized for both systemic and local drug delivery applications. This review gives an overview of long acting injectable systems that are currently in clinical use. These products are categorised in three different groups: biodegradable polymeric systems, including microparticles and implants; micro and nanocrystal suspensions and oil-based formulations. Furthermore, the applications of these drug delivery platforms for the management of various chronic diseases are summarized. Finally, this review addresses industrial challenges regarding the development of long acting injectable formulations