Rose Bengal (RB) is a photosensitizer dye used for eradicating cancer and microbial cells. Intralesional 10% RB (PV-10®) demonstrated to kill locoregional and metastatic melanoma, in the absence of light, by direct tumor ablation and indirect tumor death thanks to a specific immune response [1]. Nevertheless, RB is only employed in ophthalmic diagnostic because the biopharmaceutical profile limits its therapeutic use [2,3]. Since formulating RB delivery systems has proven to overcome its limits [4], this work focuses on the development of RB lipid-based nanocarriers to treat melanoma via topical application in the absence of light. Solid lipid nanoparticles (SLNs), obtained by solvent emulsification-evaporation, and transfersomes (TFs) obtained by modified reverse evaporation (REV), were prepared both blank and RB-loaded. Dimensional properties, physical stability and the interaction between RB and lipids were evaluated. For blank SLNs, 90 s of sonication at 70% US amplitude allowed to obtain a homogenous polydispersity index (PI=0.225) and stable formulation without phase separation or aggregation. About blank TFs, we obtained the best dimensional stability by REV technique: after three months, size increased from 244 nm to 497.35 nm while TFs prepared by another technique increased from 236.23 nm to 747.4 nm. Moreover, leader TFs were prepared employing ethanol instead of an initial toxic solvent mixture obtaining a safe and more homogeneous system (PI=0,166 vs PI=0.43). Dimensional studies on loaded nanosystems revealed SLNs were 130 nm-sized and TFs 230 nm, both were homogeneous (PI<0.25) and stable after one month. UV-spectrophotometer analysis showed RB maximum absorption wavelength shifted from 549 nm in water to 562 nm when it is formulated in carriers indicating an association with the lipid phase [5]. Cytotoxicity studies on melanoma cells are ongoing; further characterizations and permeation studies are planned. We obtained two stable RB formulations potentially active in eradicating melanoma via dermal delivery.

1. Liu, H.; Innamarato, P.P.; Kodumudi, K.; Weber, A.; Nemoto, S.; Robinson, J.L.; Crago, G.; McCardle, T.; Royster, E.; Sarnaik, A.A.; et al. Intralesional rose bengal in melanoma elicits tumor immunity via activation of dendritic cells by the release of high mobility group box 1. Oncotarget 2016, 7, 37893–37905, doi:10.18632/oncotarget.9247.
2. Klaassen, C.D. Pharmacokinetics of rose bengal in the rat, rabbit, dog, and guinea pig. Toxicol. Appl. Pharmacol.1976, 38, 85–100.
3. Pellosi, D.S.; Estevão, B.M.; Semensato, J.; Severino, D.; Baptista, M.S.; Politi, M.J.; Hioka, N.; Caetano, W. Photophysical properties and interactions of xanthene dyes in aqueous micelles. J. Photochem. Photobiol. Chem.2012, 247, 8–15, doi:10.1016/j.jphotochem.2012.07.009.
4. Loya-Castro, M.F.; Sánchez-Mejía, M.; Sánchez-Ramírez, D.R.; Domínguez-Ríos, R.; Escareño, N.; Oceguera-Basurto, P.E.; Figueroa-Ochoa, É.B.; Quintero, A.; del Toro-Arreola, A.; Topete, A.; et al. Preparation of PLGA/Rose Bengal colloidal particles by double emulsion and layer-by-layer for breast cancer treatment. J. Colloid Interface Sci.2018, 518, 122–129, doi:10.1016/j.jcis.2018.02.013.
5. Hugo, E.; Abuin, E.; Lissi, E.; Alarcón, E.; Edwards, A.M. Effect of temperature on the photobehavior of Rose Bengal associated with dipalmitoylphosphatidyl choline liposomes. J. Lumin. 2011, 131, 2468–2472, doi:10.1016/j.jlumin.2011.06.021.

Osteoarthritis (OA) is one of the most frequent degenerative joint disease characterized by joint pain and stiffness traditionally treated with symptomatic drugs like oral NSAIDs and, in extreme cases, with intra-articular corticoids. However, both these drugs are not exempt from adverse effects. Curcumin (Cur) has proven its anti-inflammatory properties and its potential as anti-osteoarthritic drug. However, its low solubility hinders its usage and limits its therapeutic efficacy. To overcome this issue, drug-in-cyclodextrin double-loaded liposomes (DCL-DL) were developed. These liposomes contained free drug in the lipid bilayer and drug-cyclodextrin complex in the aqueous compartment. The aim of this work was to evaluate the actual effectiveness of Cur-DCL-DL formulations in the OA treatment by intra-articular treatment. For this purpose, the monoiodoacetate (MIA) model of OA pain in rats was used. A single dose of samples containing Cur as DCL-DL, conventional liposomes (SL) and empty liposomes (EL, as control) were injected once intra-articularly. Paw-pressure, beam-balance and incapacitation tests were performed to evaluate OA progression at 7 and 14 days. After ending the assay, animals were sacrificed, and histological evaluation of the ankle-joint tissue was performed. Results showed that DCL-DL significantly reduced pain and ameliorated balance and gait of rats over the 14 days compared to SL. Histological tests showed that DCL-DL had protective properties in some aspects of OA.

Carbamazepine, (CBZ) is a sodium channel blocker recommended for the treatment of epilepsy and trigeminal neuralgia for over 40 years^1-3. Cyclodextrins are the most suitable candidate for the inclusion of CBZ due to the dimension of the internal cavity in which the active drug can fit properly. The aim of the current study is to evaluate the ability of β - cyclodextrin to include CBZ. The kneading method of complexation in solid state (1:1 molar ratio) was used to obtain the inclusion complex. For comparison a simple physical mixture was prepared. Physical and chemical characterization of raw materials, physical mixture and the inclusion complex were made using Fourier transform – infrared spectroscopy, X-ray diffraction, scanning electron microscopy and simultaneous thermal analysis. The results obtained using all these analytical techniques, proved that carbamazepine forms stable complexes with β-cyclodextrin in 1:1 molar ratio, and the complexation was almost complete.

[1] Bauer, J, Monika, BM., Reuber, M. Treatment strategies for focal epilepsy. Expert Opin Pharmacother. 2009;10:743–753.

[2] Goodman, LS., Gilman, AG., Hardman, JG., Limbird, LE. Gilman’s the Pharmacological Basis of Therapeutics, 10th ed. Goodman & McGraw-Hill Book Co., New York; 2001.

[3] Kwan, P., Sills, GJ., Brodie, MJ. The mechanisms of action of commonly used antiepileptic drugs. Pharmacol Ther. 2001; 90: 21-34.

Acknowledgements: This contribution was elaborated within the „Green Chemistry” research program of the “Ilie Murgulescu” Institute of Physical Chemistry of the Romanian Academy. Support of the EU (ERDF) and Romanian Government that allowed for acquisition of the research infrastructure under POS-CCEO 2.2.1 project INFRANANOCHEM – Nr. 19/01.03.2009 is gratefully acknowledged.

Chitosan is getting growing attention to increase the bioavailability of active pharmaceutical ingredients (APIs) and achieve controlled drug delivery. Through the nasal epithelium, the API absorption is rapid, which is beneficial in relieving acute pain, so intranasally administered meloxicam (MEL) could attain a rapid analgesic effect. The aims of our work were to develop spray-dried cross-linked and non-cross-linked chitosan-based drug delivery systems administrated for the intranasal route, and to optimize the spray-drying process parameters and the composition. The appropriate process parameters (inlet air temperature, pump rate) were determined based on the particle size distribution and morphology of drug-free chitosan particles. MEL-containing samples were prepared using different amounts of sodium tripolyphosphate (TPP). The micrometric properties, structural characterization and in vitro drug release were studied. Spray drying resulted in micronized chitosan particles regardless of the process parameters. The particle size of API-containing samples suited the requirements of intranasal powders, showed nearly spherical habit and MEL was present in a molecularly dispersed state in them. The highest MEL amount dissolved from the non-cross-linked MEL-containing sample. Our results indicate that spray-dried MEL-containing chitosan microparticles may be recommended for the development of a novel drug delivery system to decrease acute pain or enhance analgesia.

Most new low molecular weight chemical entities in pharmaceutical developments suffer from a low aqueous solubility, making oral delivery challenging. Despite the numerous formulation efforts that can be investigated, research especially on amorphous drugs and formulations appears to be a useful approach. Whilst few drugs can be converted to an amorphous form on their own, due to a too physical stability, the use of amorphous solid dispersions, i.e. the dissolution of drug molecules into (amorphous) polymers is increasingly used. However, certain shortcoming of these polymers based amorphous solid dispersions, such as a low drug load and a usually high hygroscopicity; still necessitate the investigation of alternative approaches. One such approach is the use of co-amorphous systems, i.e. to combination of initially crystalline low molecular weight drugs and excipients. Usually here a 1:1 molar ratio is used, but this may not be the optimal mixing ratio. In this presentation, work on investigating to optimal ratio between drug and co-former will be presented and critically discussed.

The Wasan laboratory at the University of British in conjunction with iCo Therapeutics Inc. has made significant strides toward the development of a lipid-based amphotericin B formulation for oral administration. Pre-clinical and clinical studies completed to date indicate that our oral formulations is highly efficacious and exhibits low toxicity within the dosage range required in treating diseases such as systemic fungal infections and leishmaniasis.

Each year in the Indian subcontinent alone, over 500,000 individuals play host to Leishmania donovani, an insidious parasite that invades macrophages, rapidly infiltrates the vital organs and ultimately leads to severe infection of the visceral reticuloendothelial system. Visceral leishmaniasis, also known as Kala-azar, is most prevalent in the weak and the young within a population. Left untreated, almost all infected individuals will die. The therapeutic arsenal against Leishmania is limited to a small number of parenterally administered agents, with daily injections of pentavalent antimony compound for 28 days being the usual course of action. Due to increasing resistance, antimonial drugs can no longer be used in many areas, including northeastern India where the incidence of Kala-azar is highest.

Amphotericin B is the current secondary treatment of choice against leishmaniasis and has a 97% cure rate with no reported resistance. However, therapy with the first-generation formulation (FungizoneR) involves IV administration over a period of 30 to 40 days and is associated with infusion and drug-related side-effects (infection of the indwelling catheter, patient chills and shaking due to RBC haemolysis, dose-dependent renal toxicity, fever, bone pain, thrombophlebitis).

Although lipid-based second-generation formulations exist (AbelcetR and AmBisomeR), which require a shorter course of therapy (3-5 days), are highly effective and exhibit lower toxicity when compared to FungizoneR, the cost of these formulations is a barrier to widespread use. Due to the difficult route of drug administration, toxicity issues and cost, amphotericin B is failing to reach the infected population and mortality rates continue to rise.

The development of an inexpensive, safe and effective oral treatment is paramount in order to address both early and late stages of this deadly disease and drug-resistant forms of VL. This talk will highlight our current findings and future goals.

Increasingly, the most potent emerging medicines are based not on small-molecule active ingredients but instead on large biomolecules such as peptides and proteins, or even on cells and sub-cellular components. These are difficult to process using traditional pharmaceutical processing approaches because their three-dimension tertiary structure is easily degraded, for instance by the application of heat in spray drying. Since electrospinning avoids the use of heat, it offers a potential route to develop stable and potent medicines from these active ingredients. This presentation will describe recent results in the production of electrospun formulations of protein drugs and extracellular vesicles.

Poor aqueous solubility is a pressing problem especially for oral drug delivery. Amorphous drug delivery systems present one of the most promising approaches to overcome this challenge. The inherent physical instability of pure amorphous drugs usually requires the addition of stabilizing excipients. Here, polymeric materials have been widely explored in combination with the poorly soluble drugs resulting in so called polymeric amorphous solid dispersions (ASDs). Whilst some products using the ASD technology have reached the market, the technology is often limited by low drug loadings (≤30wt%) or insufficient solubility enhancement. Recently, proteins as excipients have been introduced as amorphous stabilizers and solubility enhancers in protein based amorphous formulations, so called Dispersomes®. In particular, whey proteins enabled the formulation of physically stable Dispersomes® at drug loadings of ≥50wt%, with improved dissolution, solubility and bioavailability compared to polymeric ASDs and the crystalline drug. The presentation will give an overview on the state-of-the-art of the Disperosme® technology.

One of the areas of pharmaceutical research into oral solid dosage forms that has seen a significant surge in interest lies in the optimisation of physical forms of active pharmaceutical ingredients (APIs). Alterations of the solid form of APIs aim at improving a drug’s solubility, permeability and therefore, bioavailability. Research has led to the development of various crystalline multicomponent API systems, such as salts and co-crystals, with the recent emphasis on the manufacture of non-crystalline or weakly crystalline materials such as amorphous solid dispersions or less traditional forms such as ionic liquids and deep eutectic mixtures. To design such systems, the principles of anti-crystal engineering might be applied, whereby instead of purposely forming a crystalline, multicomponent system based on specific synthon interactions, the opposite is accomplished and a mixture with a weak or no crystallisation potential develops. For pharmaceutical applications, such a mixture will have a very desirable property – non-existent or very low crystal lattice (energy) allowing the drug molecules to be readily available for transport and absorption.

This work will concentrate on the mechanochemical, green approach to anti-crystal engineering. The concept of amorphous polymeric salts will be presented, with ball mill facilitating the proton transfer between the API and the polymer leading to an increase in solubility and biological activity of the drug. Another example will show how mechanochemistry can facilitate the formation of ionic liquids and (deep) eutectic mixtures with studies on how the counterion/co-former can manipulate the properties of the resulting phases.

Introduction. Cyclodextrins are used in various areas due to their ability to form inclusion complexes and to modify some properties of the guest molecule. The characteristics that can be improved are related to solubility of poorly water-soluble drugs, stabilization of labile guests against the degradative effects of environment (oxidation, light and heat), bioavailability, decreasing of side effects, taste modification or odour elimination and controlling of drug release. Also, ibuprofen is a very used nonsteroidal anti-inflammatory drug in treating pain and inflammation, but the long-term use of this drug has been associated with gastrointestinal side effects and nephrotoxicity. These led to introducing of new compounds of ibuprofen with improved profile. Aim. The research project combine two actual topics in pharmaceutical area: developing new safer drugs and improving the pharmacokinetic and pharmacotoxicological profile through complexation with cyclodextrins. The main objective was to develop drug delivery systems based on cyclodextrins and new ibuprofen thiazolidin-4-ones as potential analgesic and anti-inflammatory drugs. Materials and methods. Thiazolidin-4-one derivatives of ibuprofen were included in β-cyclodextrin complexes by co-precipitation (1:1M) and lyophilization methods. The inclusion complexes were characterized using spectral methods such as infrared analysis (FTIR), NMR spectroscopy and phase solubility studies. The surface morphology was studied using scanning electron microscopy (SEM). Results and conclusions. There were obtained and characterized 4 inclusion complexes with β-cyclodextrin and new ibuprofen derivatives with thiazolidin-4-one structure. These can confirm the theoretical premises for an improved pharmacological and safety toxicological profile and can continue with future studies (in vivo biological evaluation of pharmacokinetic, analgesic and anti-inflammatory profile).