Development and investigation of implantable tablets for wound healing modulation after trabeculectomy

Modulation of wound healing is required to inhibit scar formation after trabeculectomy. Mitomycin C and 5-fluorouracil (5-FU) are the anti-proliferative drugs currently used in the clinic. Their effectiveness is limited due to their toxicity and suboptimal local tissue pharmacokinetics caused by rapid clearance within the subconjunctival space. Therefore, prolonged release of anti-scarring agents is needed. Ilomastat is a matrix metalloproteinase inhibitor that has been shown to moderate favorably the wound healing response after surgery. However, due to rapid local tissue clearance, several injections are necessary in a clinically relevant animal model. The thesis describes the development and investigation of implantable ocular tablets for wound healing modulation after trabeculectomy. A tablet would be placed in the subconjunctival space called a bleb, which is caused by the outflow through the fistula made during surgery. While the bleb volume varies, the tablet is thought to dissolve in non-sink conditions. Tablet dissolution was evaluated in vitro using a flow chamber that mimics the liquid outflow conditions of the bleb. The in vitro release studies suggest that tablet dissolution would act to prolong drug release in comparison with injections. Tablet dissolution in the subconjunctival space is affected by many factors including the physical/chemical properties of the drug and tablet, temperature, the volume and flow rate of the dissolution media, and the surface area of the tablet. The efficacy of an excipient-free ilomastat tablet was evaluated using a clinically validated rabbit model. The tablet dissolution was examined using molecular dynamics simulations to consider the molecular interactions of ilomastat and 5-FU undergoing dissolution in non-sink conditions. A mathematical model was also developed to describe the dissolution of excipient-free tablets in the non-sink conditions with 94% confidence. In conclusion, we have established methods to investigate and simulate dissolution of excipientless tablets in non-sink conditions within the subconjunctival space

LC-MS analysis to determine the biodistribution of a polymer coated ilomastat ocular implant

Ilomastat is a matrix metalloproteinase inhibitor (MMPi) that has shown the potential to inhibit scarring (fibrosis) by mediating healing after injury or surgery. A long lasting ocular implantable pharmaceutical formulation of ilomastat is being developed to mediate the healing process to prevent scarring after glaucoma filtration surgery. The ilomastat implant was coated with water permeable and biocompatible phosphoryl choline polymer (PC1059) displayed extended slow release of ilomastat in vitro and in vivo. The ocular distribution of ilomastat from the implant in rabbits at day 30 post surgery was determined by the extraction of ilomastat and its internal standard marimastat from the ocular tissues, plasma, aqueous humour and vitreous fluid followed by capillary-flow liquid chromatography (cap-LC), the column effluent was directed into a triple quadrupole mass spectrometer operating in product scan mode. The lower limits of quantification (LLOQs) were 0.3 pg/μL for ocular fluids and plasma, and 3 pg/mg for ocular tissues. The extraction recoveries were 90-95% for ilomastat and its internal standard from ocular tissues. Ilomastat was found in ocular fluids and tissues at day 30 after surgery. The level of ilomastat was 18 times higher in the aqueous humour than vitreous humour. The concentration ranking of ilomastat in the ocular tissues was sclera > bleb conjunctiva > conjunctiva (rest of the eye) > cornea. Mass spectrometry analysis to confirm the presence of ilomastat in the ocular tissues and fluids at day 30 post-surgery establishes the extended release of ilomastat can be achieved in vivo, which is crucial information for optimisation of the ilomastat coated implant

Integration of advanced methods and models to study drug absorption and related processes : An UNGAP perspective

Funding Information: AI acknowledges the support of projects icp009 (ALKOOL) of PRACE-ICEI (grant agreement 800858) for awarding access to Piz Daint, at the Swiss National Supercomputing Centre (CSCS), Switzerland and BG05M2OP001–1.001–0004 (UNITe) of the Bulgarian Ministry of Education and Science. For further details on points raised in this article, please contact [email protected]. Funding Information: Acknowledgements. JAGH is supported by the Biocenter Finland, the Helsinki Institute of Life Sciences, and the Faculty of Pharmacy, University of Helsinki. Publisher Copyright: © 2021 The AuthorsThis collection of contributions from the European Network on Understanding Gastrointestinal Absorption-related Processes (UNGAP) community assembly aims to provide information on some of the current and newer methods employed to study the behaviour of medicines. It is the product of interactions in the immediate pre-Covid period when UNGAP members were able to meet and set up workshops and to discuss progress across the disciplines. UNGAP activities are divided into work packages that cover special treatment populations, absorption processes in different regions of the gut, the development of advanced formulations and the integration of food and pharmaceutical scientists in the food-drug interface. This involves both new and established technical approaches in which we have attempted to define best practice and highlight areas where further research is needed. Over the last months we have been able to reflect on some of the key innovative approaches which we were tasked with mapping, including theoretical, in silico, in vitro, in vivo and ex vivo, preclinical and clinical approaches. This is the product of some of us in a snapshot of where UNGAP has travelled and what aspects of innovative technologies are important. It is not a comprehensive review of all methods used in research to study drug dissolution and absorption, but provides an ample panorama of current and advanced methods generally and potentially useful in this area. This collection starts from a consideration of advances in a priori approaches: an understanding of the molecular properties of the compound to predict biological characteristics relevant to absorption. The next four sections discuss a major activity in the UNGAP initiative, the pursuit of more representative conditions to study lumenal dissolution of drug formulations developed independently by academic teams. They are important because they illustrate examples of in vitro simulation systems that have begun to provide a useful understanding of formulation behaviour in the upper GI tract for industry. The Leuven team highlights the importance of the physiology of the digestive tract, as they describe the relevance of gastric and intestinal fluids on the behaviour of drugs along the tract. This provides the introduction to microdosing as an early tool to study drug disposition. Microdosing in oncology is starting to use gamma-emitting tracers, which provides a link through SPECT to the next section on nuclear medicine. The last two papers link the modelling approaches used by the pharmaceutical industry, in silico to Pop-PK linking to Darwich and Aarons, who provide discussion on pharmacometric modelling, completing the loop of molecule to man.Peer reviewe

Degrees of order : A comparison of nanocrystal and amorphous solids for poorly soluble drugs

Poor aqueous solubility is currently a prevalent issue in the development of small molecule pharmaceuticals. Several methods are possible for improving the solubility, dissolution rate and bioavailability of Biopharmaceutics Classification System (BCS) class II and class IV drugs. Two solid state approaches, which rely on reductions in order, and can theoretically be applied to all molecules without any specific chemical prerequisites (compared with e.g. ionizable or co-former groups, or sufficient lipophilicity), are the use of the amorphous form and nanocrystals. Research involving these two approaches is relatively extensive and commercial products are now available based on these technologies. Nevertheless, their formulation remains more challenging than with conventional dosage forms. This article describes these two technologies from both theoretical and practical perspectives by briefly discussing the physicochemical backgrounds behind these approaches, as well as the resulting practical implications, both positive and negative. Case studies demonstrating the benefits and challenges of these two techniques are presented.Peer reviewe

Nanocrystal formulation for poorly soluble drugs

Poorly soluble drugs are often a challenging problem in drug formulation. Reducing the particle size of the drug to a nano-scale leads to an increased surface area-to-volume ratio, increased dissolution velocity and adhesiveness, and improved in vivo performance of poorly soluble drugs. Wet media milling is one of the most popular techniques to prepare the nanocrystals. The aim of this thesis was to optimize the preparation conditions and characterization methods of nanosuspensions for poorly water-soluble drug compounds, then formulate the nanosuspensions into suitable pharmaceutical dosage forms, and finally test the efficacy in in vivo. Firstly, basic research on the preparation and characterization of nanosuspensions was done using the wet milling technique and poorly water-soluble drugs as model compounds. The various milling parameters, including the milling time, diameter of milling balls, stabilizer type and concentrations, were investigated. The particle size, size distribution and stability of nanosuspensions were considered as the important parameters to evaluate the milling conditions. The drug nanocrystals manufactured at the optimal milling environment exhibited significantly increased dissolution rates, good stability and kept a crystalline state. Secondly, the importance of particle size on dissolution was determined. The conventional dissolution method can effectively discriminate the milled and unmilled samples, but it fails to discriminate the dissolution profiles of nanosuspensions with different particle sizes. A novel condition, i.e. the sample concentration in the dissolution medium is close to the apparent saturation solubility of the drug, were found to discriminate the dissolution profiles of nanosuspensions. Thirdly, nanos-in-micros structure for suitable pharmaceutical formulations, e.g. tablet or pulmonary products was set up. The structure is that drug nanocrystals are buried inside microparticle carriers, in which the mannitol and crystalline L-leucine were as matrix and outer layer formers, respectively. Nanos-in-micros structure avoids problems in further formulation of nanopowders, such as poor flowability and aggregation/sinter, meanwhile functionality of individual nanoparticles in microparticles are remained, such as fast dissolution. Finally, the efficacy of nanocrystal formulations in vivo was tested. Brinzolamide nanosuspensions were prepared in buffers and formulated for ocular administration in order to reduce the intraocular pressure in rats. The nanocrystal formulations exhibited a low cytotoxicity and significant reduction in intraocular pressure compared to the physiologic salt solution and untreated group. In conclusion, the nanosuspensions of poorly water-soluble drug can be easily produced by the wet milling technique when the choice of stabilizers and milling parameters are appropriate. The nanosuspensions were successfully formulated and exhibited a good in vivo performance. More efficient formulations based on drug nanosuspensions should be further researched and developed.Uusien lääkeainemolekyylien huono liukoisuus on tämän päivän lääkekehityksen suurimpia haasteita. Lääkeaineen partikkelikoon pienentäminen nanometrikokoluokkaan, nanokiteytys, on yksi tapa parantaa liukoisuusnopeutta. Partikkelikokoa voidaan pienentää esimerkiksi jauhamalla kiinteää lääkeainetta vesilietteessä, korkeapainehomogenisoinnilla tai erilaisilla saostaus ja sumutustekniikoilla. Tässä väitöskirjatyössä valmistettiin lääkeainenanokiteitä kosteajauhatusprosessilla. Aluksi optimoitiin jauhatusprosessia siten, että saatiin tuotettua mahdollisimman hyvät liukoisuusominaisuudet omaavia ja hyvin säilyviä nanokiteitä. Myös nanokiteiden stabiloinnissa käytettäviä erilaisia apuaineita ja niiden tehokkuutta lääkeainenanokiteiden stabiloinnissa tutkittiin. Nanokiteiden tarkkaa liukoisuustutkimusta varten kehitettiin olemassa olevia liukoisuuskoetestejä herkempi analysointitekniikka. Pulmonaarista ja oraalista annostelua varten nanokiteistä tehtiin nanorakenteisia mikropartikkeleita uudella tekniikalla. Mikropartikkeleissa ja niistä tehdyissä lääkevalmisteissa saatiin säilytettyä nanokiteiden hyvät liukoisuusominaisuudet. Väitöstyön viimeisessä osakokonaisuudessa nanokiteistä valmistettiin silmälääkevalmisteita, ja valmisteiden toimivuus varmistettiin in vivo -kokeilla. In vivo -kokeissa kaikkien brinzolamidinanokideformulaatioiden todettiin laskevan tehokkaasti silmänpainetta. Väitöskirjatutkimuksen tuloksena saatiin uutta tietoa jauhatusprosessin parametrien ja apuainevalintojen tekemiseen lääkeainenanokiteiden valmistuksessa. Lisäksi työssä kehitettiin uusi nanorakenteisten mikropartikkelien valmistustapa sekä tarkempaa analytiikkaa. In vivo -kokeiden perusteella saatiin vahvistettua valmisteiden toimivuus myös koe-eläimillä

Pharmaceutical Particulates and Membranes for Delivery of Drugs and Bioactive Molecules

This book is a collection of papers published in the Special Issue of Pharmaceutics, entitled "Pharmaceutical Particulates and Membranes for Delivery of Drugs and Bioactive Molecules". A drug release profile is a consequential factor for nanoparticle application, directly related to drug stability and therapeutic results, as well as formulation development. Pharmaceutical particulates of different sizes and shapes (e.g., liposomes, oil-in-water emulsions, polymeric nano- and microspheres, metallic nanoparticles (NPs) such as gold, silver and iron oxide crystals, and core-shell hybrid NPs) offer many diagnostic and therapeutic applications. Membranes are also extensively utilized in many applications. They are especially beneficial to the distribution of macromolecular drugs and biopharmaceutical drugs (peptides, proteins, antibodies, oligonucleotides, plasmids, and viruses) with physicochemical and pharmacokinetic vulnerability. The delivery of drugs and bioactive molecules using particulates and membranes has gained a great deal of attention for various applications, such as the treatment of secondary infections, cancer treatment, skin regeneration, orthopaedic applications, and antimicrobial drug delivery. In addition, several production techniques have been utilized for the fabrication of particulates and membranes in the last decade, which include lyophilisation, micro-emulsion, nano-spray dryer, nano-electrospinning, slip casting and 3D printers. Therefore, pharmaceutical particulates and membranes possess excellent prospects to deliver drugs and bioactive molecules with the potential to improve new delivery strategies like sustained and controlled release

In vitro and in vivo delivery of a sustained release nanocarrier-based formulation of an MRTF/SRF inhibitor in conjunctival fibrosis

Abstract Background Sustained drug delivery is a large unmet clinical need in glaucoma. Here, we incorporated a Myocardin-Related Transcription Factor/Serum Response Factor inhibitor, CCG-222740, into slow release large unilamellar vesicles derived from the liposomes DOTMA (1,2-di-O-octadecenyl-3-trimethylammonium propane) and DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine), and tested their effects in vitro and in vivo. Results The vesicles were spherical particles of around 130 nm and were strongly cationic. A large amount of inhibitor could be incorporated into the vesicles. We showed that the nanocarrier CCG-222740 formulation gradually released the inhibitor over 14 days using high performance liquid chromatography. Nanocarrier CCG-222740 significantly decreased ACTA2 gene expression and was not cytotoxic in human conjunctival fibroblasts. In vivo, nanocarrier CCG-222740 doubled the bleb survival from 11.0 ± 0.6 days to 22.0 ± 1.3 days (p = 0.001), decreased conjunctival scarring and did not have any local or systemic adverse effects in a rabbit model of glaucoma filtration surgery. Conclusions Our study demonstrates proof-of-concept that a nanocarrier-based formulation efficiently achieves a sustained release of a Myocardin-Related Transcription Factor/Serum Response Factor inhibitor and prevents conjunctival fibrosis in an established rabbit model of glaucoma filtration surgery.https://deepblue.lib.umich.edu/bitstream/2027.42/146540/1/12951_2018_Article_425.pd

An Ilomastat-CD Eye Drop Formulation to Treat Ocular Scarring

PURPOSE: The purpose of this study was to develop a topical matrix metalloproteinase inhibitor preparation for antiscarring therapy. METHODS: The broad spectrum matrix metalloproteinase inhibitor ilomastat was formulated using 2-hydroxypropyl-β-cyclodextrin in aqueous solution. In vitro activity of ilomastat-cyclodextrin (ilomastat-CD) was examined using fibroblasts seeded in collagen. Permeation of ilomastat-CD eye drop through pig eye conjunctiva was confirmed using Franz diffusion cells. Ilomastat-CD eye drop was applied to rabbit eyes in vivo, and the distribution of ilomastat in ocular tissues and fluids was determined by liquid chromatography-mass spectroscopy. RESULTS: The aqueous solubility of ilomastat-CD was ∼1000 μg/mL in water and 1400 μg/mL in PBS (pH 7.4), which is greater than ilomastat alone (140 and 160 μg/mL in water and PBS, respectively). The in vitro activity of ilomastat-CD to inhibit collagen contraction in the presence of human Tenon fibroblast cells was unchanged compared to uncomplexed ilomastat. Topically administered ilomastat-CD in vivo to rabbit eyes resulted in a therapeutic concentration of ilomastat being present in the sclera and conjunctiva and within the aqueous humor. CONCLUSIONS: Ilomastat-CD has the potential to be formulated as an eye drop for use as an antifibrotic, which may have implications for the prevention of scarring in many settings, for example glaucoma filtration surgery