Sustained Release Ocular Bevacizumab for the treatment of wet acute macular degeneration This feasibility study is to verify the technical and economic viability of a sustained release bevacizumab product (SR Bevacizumab) to improve the safety, cost-effectiveness and patient acceptability for the treatment of wet age-related macular degeneration (AMD). AMD is the leading cause of blindness in those aged over 50 and causes blurring, distortion and loss of central vision and almost always affects quality of life. Prototype SR Bevacizumab products will be manufactured using CriticalMix, an advanced manufacturing and processing technology developed by Critical Pharmaceuticals, which will encapsulate bevacizumab in biodegradable polymers suitable for sustained release of the drug in the eye.

The project will evaluate a novel absorption promoter system - CriticalSorb for the nasal delivery of drugs. The project will focus on four key areas: evaluation of the physico-chemical properties of the formulations and identify essential characteristics, investigate its mechanisms of action using appropriate cell culture models, identify synergistic effects with other known absorption promoters and the final part of the project will look at developing a robust in vitro cell culture model in order to replace animal models when evaluating the nasal absorption of drugs. Background The nasal route of delivery can be exploited for the systemic delivery of drugs such as small molecular weight polar drug, peptides and proteins that are not easily administered via other routes than by injection. Despite the large surface area of the nasal cavity and extensive blood supply absorption of polar molecules, peptides and proteins is low but can be greatly improved if administered in conjunction with an absorption promoting agent. The enhancer systems work by a number of mechanisms however; animal studies have shown that most often there is a direct correlation between the absorption enhancing effect obtained and the damage caused to the nasal membrane. It is therefore important to discover and evaluate new absorption promoter systems. Critical Pharmaceuticals is a small specialty pharmaceutical company and have developed a nasal drug delivery platform based on an absorption promoter called CriticalSorb which has shown to be non-toxic to the nasal membrane in repeat dosing studies Task1. Physico-chemical characterisation of the formulations The first stage of the project will evaluate at the physico-chemical characterization of formulations containing CriticalSorb and a model drug in the form of a peptides or a protein. The CriticalSorb aqueous solutions form micelles and experiments will be carried out to determine the position of drug within such the micellar solutions, as well as the micellar properties. It will apply a combination of routine and state of the art 'nanotechnology' analytical techniques including: particle size and distribution, dye incorporation, morphology (cryo-TEM). Task 2. Evaluation of CriticalSorb mechanism of action The second stage of the project will study and describe the mechanism of action of the absorption enhancing effect of CriticalSorb and its principal components. This will focus on cell culture monolayers of either Calu-3 cells or HBEC (human bronchial epithelial) cells lines and will assess the mechanism of transport across the mucosal membrane, potentially transcellular or paracellular route. Specific mechanism of action will be investigated, such as the tight junction opening, clathrin or calveolin cellular pathway and effect on the 170 kDa membrane bound P-glycoprotein. Task 3. Potential synergistic effects of CriticalSorb with other known absorption enhancers CriticalSorb will be combined with other known absorption enhancers with alternative modes of action such as the typically investigated bioadhesive polysaccharide Chitosan to determine potential synergistic effects between the enhancers. Task 4. In vitro - in vivo correlation of absorption enhancing effect The final stage of the project will include comparisons of absorption enhancing effect from in vitro cell culture models with the in vivo data presently available, and those that will be created in animal studies by Critical Pharmaceuticals through another project. Wagner-Nelson modeling of the data will be carried out to determine the in vitro-in vivo correlation and the suitability of the in vitro cell culture model.

To exert their effect biopharmaceuticals require the delivery of the native biologically active molecule to the site of action. However their labile nature and poor bioavailability often presents considerable challenges to their formulation into medicines and can obstruct development and commercialisation. Furthermore, their frequently unfavourable pharmacokinetics means that they are usually administered by frequent injections which is inconvenient for patients and can effect their compliance with the therapeutic regimen. One strategy to overcome some of these issues is to develop longer acting versions of the drug. Encapsulation of the protein within a matrix which slowly releases the drug is an attractive strategy to achieve this since this does not involve any chemical modification of the drug. However to date the manufacturing processes used to achieve this have suffered a number of draw backs when applied to biopharmaceuticals. The high temperatures used in various polymer melt processes are known to denature proteins causing them to lose their activity. Alternatively, emulsion processes can generate aqueous-organic interfaces with a very large surface area at which proteins can also denature. Furthermore, these emulsion processes are associated with the economic and environmental cost of solvent disposal. To date Nutropin Depot (a controlled release version of human growth hormone (hGH)) has been the only protein encapsulated within a polymeric matrix to successfully reach the market. This was manufactured using a proprietary emulsion process but was eventually withdrawn because it was reported that the manufacturing process was 'too resource intensive'. Critical Pharmaceuticals is a small specialty pharmaceutical company, with an innovative encapsulation technology using supercritical fluids (SCF). We have previously demonstrated the potential of SCF processing to encapsulate drugs within polymer microparticles in a single step process. This technology has the advantage that it operates at ambient temperatures and in the absence of solvents, permitting the formulation of biopharmaceuticals without adversely affecting their activity. We are currently developing the technology with the aim of gaining regulatory acceptance and are on track to initiate a phase 1 clinical trial in 2008 for our lead product, a controlled release protein formulation. The product properties need to be tailored to the specific application in order to successfully take these products to market, and aid downstream processing. For example, depending upon the application the particle size, shape and flow properties of the products needs to be tightly defined e.g. in order to aid injectability or vial filling. With this CASE award we hope to build on recent advances made both at the University of Nottingham and in Critical Pharmaceuticals' laboratories that have optimised the supercritical apparatus to allow the reproducible manufacture of polymer microparticles containing encapsulated biopharmaceuticals. These products have been shown to control the release of the drug both in vitro and in vivo. We have discovered that the nature of the products produced is influenced not only by the polymeric material itself and conditions that it is processed at, but also that the addition of excipients can impact upon particle size, morphology and performance. We have therefore identified a complex interplay between the SCF process and the formulation which impacts upon the products produced - their properties, performance and applicability. In this project we propose to apply experimental design to determine the key processing and formulation parameters which impact upon particle size, shape, surface area, flow, porosity and performance, with the aim of gaining a greater understanding how each of the variables interact. This will lead to the production of protein formulations with enhanced performance and ultimately help improve patient lives.

Supercritical fluid technology for gastroretention formulations Brief description of proposed project: The project applies supercritical fluid technology to the design of highly porous micron-sized particles capable of gastric retention. The process parameters and microparticles properties required to achieve gastroretention, and improve oral delivery of 'problematic' drugs, will be investigated. Background Drugs with an absorption window in the upper small intestine have limited bioavailability when applied as conventional oral dosage forms, as the residence time of these formulations in the stomach and upper intestine is short, hence limiting extend of the drug absorption. To increase the bioavailability of the 'absorption window' drugs, formulations which the prolong residence time have been suggested, including (i) systems that adhere to the mucosal surface, (ii) systems that rapidly increase in size and thus cannot leave the stomach and (iii) systems with optimized density to float on gastric fluids (1-2). These systems are usually based on the 'one unit' approach but their reliability (due to such a design) has been questioned and application of 'multiple unit systems' (3) such as floating capsules with granules of gas generating agents (3) or hollow microparticles (4) has been suggested. We believe that the application of supercritical fluid (SCF) technology will make a step change in the hollow particles technology, compared to the current double emulsion (eg. 1,4) as, via changes in the kinetics of carbon dioxide depressurisation, one can generate formulations of variable densities, sizes and morphologies. The technology is generic and potential applications are in eg. systemic therapy of Alzhemier disease (L-dopa), CNS therapy (diazepam), cardiovascular therapy (propranolol), pulmonary oedema (furosemid) or local therapy of eg Helicobacter pylori (5). Project plan Task1. Preparation of hollow microparticles by SCF technology Initially the project will select materials and conditions to produce microparticles with controlled size ranges, densities and morphologies. Materials will initially be based on lactic acid and polyethhylne oxide polymers and their combinations, as established within the company. Processing of other materials will also be investigated, such as combinations of phospholipids and fatty acids. Task 2. In vitro characterization and drug incorporation The in vitro characterization aims to establish a fundamental understanding of a correlation between the buoyancy of microparticles (in simulated gastric conditions) and their physicochemical properties: size, density (and internal structure) and morphology (eg roundness). The drug loading potential of the microparticles for model drug compounds (L-dopa, furosemid and indomethacin) will be investigated and correlated to the nature of the material and the processing conditions. The release profile of incorporated compounds will be determined and optimized to correlate with the potential gastric retention time. Task 3. In vivo performance A limited number of selected systems will be investigated for their in vivo performance. These will include a dosing of experimental animals (pig) to determine plasma concentration profiles (bioavailability). The studies will be conducted in collaboration Prof. J Wiseman, School of Biosciences, Univ. of Nottingham. There is a current established collaboration in animal experimentation on the mucosal delivery and a DTI Technology Programme project (D Gray) to which both J Wiseman and S Stolnik are contributors. References 1. Streubel A et al. Current Opinion in Pharmacology 6 (2006) 501 2. Brahma N et al. Journal Controlled Release 63 (2000) 235. 3 Shweta Arora et al. AAPS PharmSciTech 6 ( 2005) Art. 47 (http://www.aapspharmscitech.org). 4 Yasunori Satoa et al, Journal of Pharmaceutics and Biopharmaceutics 55 (2003) 297. 5 Bardonnet PL et al. Journal Controlled Release 111 (2006)

Regenerative medicine (RM) is a convergence of conventional pharmaceutical sciences, medical devices and surgical intervention employing novel cell and biomaterial based therapies. RM products replace or regenerate damaged or defective tissues such as skin, bone, and even more complex organs, to restore or establish normal function. They can also be used to improve drug testing and disease modelling. RM is an emerging industry with a unique opportunity to contribute to the health and wealth of the UK. It is a high value science-based manufacturing industry whose products will reduce the economic and social impact of an aging population and increasing chronic disease.The clinical and product opportunities for RM have become clear and a broad portfolio of products have now entered the translational pipeline from the science bench to commercialisation and clinical application. The primary current focus for firms introducing these products is first in man studies; however, success at this stage is followed by a requirement for a rapid expansion of delivery capability - the 'one-to-many' translation process. This demands increasing attention to regulatory pathways, product reimbursement and refinement of the business model, a point emphasised by recent regulatory decisions demanding more clarity in the criteria that define product performance, and regulator initiatives to improve control of manufacturing quality. The IMRC will reduce the attrition of businesses at this critical point in product development through an industry facing portfolio of business driven research activities focussed on these translational challenges. The IMRC will consist of a platform activity and two related research themes. The platform activity will incorporate studies designed to influence public policy, regulation and the value system; to explore highly speculative and high value ideas (particularly clinically driven studies); and manufacturing-led feasibility and pilot studies using state of the art production platforms and control. The research themes will focus on areas identified as particular bottlenecks in RM product translation. The first theme will explore the delivery, manufacturing and supply processes i.e. the end to end production of an RM product. Specifically this theme will explore using novel pharmaceutical technology to control the packaged environment of a living RM product during shipping, and the design of a modular solution for manufacturing different cell based therapies to the required quality in a clinical setting. The second research theme will apply quality by design methods to characterise the quality of highly complex RM products incorporating cells and carrier materials. In particular it will consider optical methods for non-invasive process and product quality control and physicochemical methods for process monitoring.The IMRC will be proactively managed under the direction of a Board and Liaison Group consisting of leading industrialists to ensure that the Centre delivers maximum value to the requirements of the business model and assisting the growth of this emerging industry.