International Journal of Pharmaceutics (v.314, #2)

Local controlled drug delivery to the brain by Juergen Siepmann (99-100).

The mass transport mechanisms involved in the controlled delivery of drugs to living brain tissue are complex and yet not fully understood. Often the drug is embedded within a polymeric or lipidic matrix, which is directly administered into the brain tissue, that is, intracranially. Different types of systems, including microparticles and disc- or rod-shaped implants are used to control the release rate and, thus, to optimize the drug concentrations at the site of action in the brain over prolonged periods of time. Most of these dosage forms are biodegradable to avoid the need for the removal of empty remnants after drug exhaustion. Various physical and chemical processes are involved in the control of drug release from these systems, including water penetration, drug dissolution, degradation of the matrix and drug diffusion. Once the drug has been released from the delivery system, it has to be transported through the living brain tissue to the target site(s). Again, a variety of phenomena, including diffusion, drug metabolism and degradation, passive or active uptake into CNS tissue and convection can be of importance for the fate of the drug. An overview is given of the current knowledge of the nature of barriers to free access of drug to tumour sites within the brain and the state of the art of: (i) mathematical modeling approaches describing the physical transport processes and chemical reactions which can occur in different types of intracranially administered drug delivery systems, and of (ii) theories quantifying the mass transport phenomena occurring after drug release in the living tissue. Both, simplified as well as complex mathematical models are presented and their major advantages and shortcomings discussed. Interestingly, there is a significant lack of mechanistically realistic, comprehensive theories describing both parts in detail, namely, drug transport in the dosage form and in the living brain tissue. High quality experimental data on drug concentrations in the brain tissue are difficult to obtain, hence this is itself an issue in testing mathematical approaches. As a future perspective, the potential benefits and limitations of these mathematical theories aiming to facilitate the design of advanced intracranial drug delivery systems and to improve the efficiency of the respective pharmacotherapies are discussed.
Keywords: Controlled release; Brain; Mathematical modeling; Transport mechanisms; Diffusion;

Several compounds that exhibit a therapeutic effect in experimental models of neurodegenerative diseases have been identified over recent years. Safe and effective drug delivery to the central nervous system is still one of the main obstacles in translating these experimental strategies into clinical therapies. Different approaches have been developed to enable drug delivery in close proximity to the desired site of action. In this review, we describe biodegradable polymeric systems as drug carriers in models of neurodegenerative diseases. Biomaterials described for intracerebral drug delivery are well tolerated by the host tissue and do not exhibit cytotoxic, immunologic, carcinogenic or teratogenic effects even after chronic exposure. Behavioral improvement and normalization of brain morphology have been observed following treatment using such biomaterials in animal models of Parkinson's, Alzheimer's and Huntington's diseases. Application of these devices for neuroactive drugs is still restricted due to the relatively small volume of tissue exposed to active compound. Further development of polymeric drug delivery systems will require that larger volumes of brain tissue are targeted, with a controlled and sustained drug release that is carefully controlled so it does not cause damage to the surrounding tissue.
Keywords: Biomaterials; Biocompatibility; Microspheres; Nanoparticles; Rods; Neurodegenerative diseases;

Paclitaxel-loaded microparticles and implants for the treatment of brain cancer: Preparation and physicochemical characterization by K. Elkharraz; N. Faisant; C. Guse; F. Siepmann; B. Arica-Yegin; J.M. Oger; R. Gust; A. Goepferich; J.P. Benoit; J. Siepmann (127-136).
The aim of this study was to prepare different types of paclitaxel-loaded, PLGA-based microparticles and lipidic implants, which can directly be injected into the brain tissue. Releasing the drug in a time-controlled manner over several weeks, these systems are intended to optimize the treatment of brain tumors. The latter is particularly difficult because of the blood–brain barrier (BBB), hindering most drugs to reach the target tissue upon systemic administration. Especially paclitaxel (being effective for the treatment of ovarian, breast, lung and other cancers) is not able to cross the BBB to a notable extent since it is a substrate of the efflux transporter P-glycoprotein. Both, biodegradable microparticles as well as small, cylindrical, glycerol tripalmitate-based implants (which can be injected using standard needles) were prepared with different paclitaxel loadings. The effects of several formulation and processing parameters on the resulting drug release kinetics were investigated in phosphate buffer pH 7.4 as well as in a diethylnicotinamide (DENA)/phosphate buffer mixture. Using DSC, SEM, SEC and optical microscopy deeper insight into the underlying drug release mechanisms could be gained. The presence of DENA in the release medium significantly increased the solubility of paclitaxel, accelerated PLGA degradation, increased the mobility of the polymer and drug molecules and fundamentally altered the geometry of the systems, resulting in increased paclitaxel release rates.
Keywords: Paclitaxel; Microparticle; Implant; Controlled release; Brain tumor;

Drug release from lipid-based implants: Elucidation of the underlying mass transport mechanisms by C. Guse; S. Koennings; F. Kreye; F. Siepmann; A. Goepferich; J. Siepmann (137-144).
The aim of this study was to better understand the mass transport mechanisms involved in the control of drug release from lipid-based implants. Different types of triglyceride-based cylinders were prepared by compression. Glycerol-trilaurate, -trimyristate, -tripalmitate and -tristearate were used as model lipids, lysozyme and pyranine as model drugs. The effects of several formulation and processing parameters on the resulting drug release kinetics in phosphate buffer pH 7.4 were studied and the obtained results analyzed using Fick's second law of diffusion. Interestingly, lysozyme release from implants prepared by compression of a lyophilized emulsion (containing dissolved drug and lipid) was found to be purely diffusion-controlled, irrespective of the type of triglyceride. In contrast, the dominating release mechanism depended on the type of lipid in the case of pyranine-loaded implants prepared by compression of a lyophilized lipid-drug solution: with glycerol-trilaurate and -tristearate the systems were found to be purely diffusion-controlled, whereas also other mass transport phenomena are of importance in glycerol-trimyristate and -tripalmitate-based devices. Similarly, changes in the size of the compressed lipid-drug particles, drug loading and compression force significantly affected the underlying release mechanisms. The addition of a drug-free, poly(lactic-co-glycolic acid) (PLGA)-based coating around the implants delayed the onset of pyranine release for about 20 days. Interestingly, the subsequent drug release was purely diffusion-controlled, irrespective of the type of triglyceride. Also the addition of different amounts (and particle size fractions) of saccharose to pyranine-loaded implants led to purely diffusion-controlled drug release.
Keywords: Lipid; Implant; Controlled release; Release mechanism; Mathematical modeling;

In vitro investigation of lipid implants as a controlled release system for interleukin-18 by S. Koennings; E. Garcion; N. Faisant; P. Menei; J.P. Benoit; A. Goepferich (145-152).
Operating on the inductive and effective phases of an anti-tumor immune response and uncovering pivotal functions that may reduce cancer cell growth, interleukin-18 (IL-18) appears to be an attractive candidate for the sustained local adjuvant immunotherapeutic treatment of brain gliomas. The objective of this work was to develop IL-18 loaded lipid implants as a controlled delivery system. For the preparation of protein loaded triglyceride matrix material, a solid-in-oil (s/o) dispersion technique was chosen for which protein particles in the micrometer range were first prepared by co-lyophilization with polyethylene glycol (PEG). Implants of 1 mm diameter, 1.8 mm height and 1.8 mg weight were manufactured by compression of the powder mixture in a specially designed powder compacting tool. The in vitro release behavior of 125I-Bolton-Hunter-radiolabeled IL-18 was assessed in a continuous-flow system. A cell culture assay was established for the determination of bioactivity of released IL-18. Implants showed a continuous release of 10–100 ng IL-18 per day for 12 days. A progressive integrity loss was observed with ongoing release, which would be related to protein degradation during incubation. The initially released fraction proved complete retention of bioactivity, indicating that the manufacturing procedure had no detrimental effects on protein stability.
Keywords: Lipid implants; Protein release; Polyethylene glycol; Interleukin-18; Biological activity assay;

Biocompatibility and erosion behavior of implants made of triglycerides and blends with cholesterol and phospholipids by C. Guse; S. Koennings; A. Maschke; M. Hacker; C. Becker; S. Schreiner; T. Blunk; T. Spruss; A. Goepferich (153-160).
Triglycerides are a promising class of material for the parenteral delivery of drugs and have become the focus of tremendous research efforts in recent years. The aim of this study was to investigate the biocompatibility of glyceroltripalmitate as well as the influence of cholesterol and distearoyl-phosphatidyl-choline (DSPC) on the erosion behavior of the lipid. For these investigations, two in vivo studies were carried out, in which cylindrical matrices of 2 mm diameter were manufactured and subcutaneously implanted in immunocompetent NMRI-mice. After excision of the implants, tissue reactions of the animals as well as changes in the weight, shape and microstructure of the implants were investigated. The triglyceride and cholesterol showed good biocompatibility, as indicated by their minimal encapsulation in connective tissue and the absence of inflammatory reactions. Increasing the levels of phospholipid in the implants, however, led to an increased inflammatory reaction. In contrast to cholesterol, which did not affect erosion, the incorporation of DSPC into the triglyceride matrices led to clearly visible signs of degradation.
Keywords: Implants; Biocompatibility; Triglyceride; Phospholipid; Cholesterol;

Programmable implants—From pulsatile to controlled release by C. Guse; S. Koennings; T. Blunk; J. Siepmann; A. Goepferich (161-169).
The aim of this study was to develop programmable implants with a reproducible delayed onset of release followed by several weeks of controlled release. For this purpose, a drug-loaded core was embedded into a drug-free bulk-eroding poly(d,l lactic-co-glycolic acid) or poly(d,l lactic acid) mantle. The manufacturing procedure was established and optimized for three mantle materials, which showed delay times ranging from 7 to 83 days. Triglycerides with fatty acid chain lengths from C12 to C18 were investigated as core materials, producing release periods from 2 to 16 weeks. Concomitantly, applying a convolution/deconvolution model showed the possibility of theoretical prediction of the resulting release profiles.
Keywords: Implants; Controlled release; Pulsatile release; Triglyceride; Convolution;

Lipidic implants for controlled release of bioactive insulin: Effects on cartilage engineered in vitro by B. Appel; A. Maschke; B. Weiser; H. Sarhan; C. Englert; P. Angele; T. Blunk; A. Göpferich (170-178).
Controlled release systems for growth factors and morphogens are potentially powerful tools for the engineering or the treatment of living tissues. However, due to possible instabilities of the protein during manufacture, storage, and release, in the development of new release systems it is paramount to investigate into the maintenance of bioactivity of the protein. Within this study, recently developed protein releasing lipid matrix cylinders of 2 mm diameter and 2 mm height made from glycerol tripalmitate were manufactured in a compression process without further additives. Insulin in different concentrations (0.2%, 1%, and 2%) served as model protein. The bioactivity of the protein released from the matrices was investigated in a long-term cartilage engineering culture for up to four weeks; additionally, the release profiles were determined using ELISA. Insulin released from the matrices increased the wet weights of the cartilaginous cell-polymer constructs (up to 3.2-fold), the amount of GAG and collagen in the constructs (up to 2.4-fold and 3.2-fold, respectively) and the GAG and collagen content per cell (1.8-fold and 2.5-fold, respectively), compared to the control. The dose-dependent effects on tissue development correlated well with release profiles from the matrices with different insulin loading. In conclusion, the lipid matrices, preserving the bioactivity of incorporated and released protein, are suggested as a suitable carrier system for use in tissue engineering or for the localized treatment of tissues with highly potent protein drugs such as used in the therapy of brain cancer or neurodegenerative CNS diseases.
Keywords: Protein; Insulin; Controlled release; Lipid; Bioactivity; Cartilage; Tissue engineering;

Development and characterization of interleukin-18-loaded biodegradable microspheres by F. Lagarce; E. Garcion; N. Faisant; O. Thomas; P. Kanaujia; P. Menei; J.P. Benoit (179-188).
Immunostimulation represents a promising approach designed to specifically eradicate malignant cells. Since glioma tumour cells hole up in the central nervous system (CNS) in a particularly inauspicious milieu to antitumour immune reactions we here propose a new strategy to revert the properties of this microenvironment by administering an antitumour cytokine into the CNS tumour itself. Thus, biodegradable poly(d,l-lactide-co-glycolide) (PLGA) sustained-release microspheres for stereotaxic implantation loaded with interleukin-18 (IL-18), that is known to exert antitumour activity and trigger immune cell-mediated cytotoxicity, were developed. Different tests for assessing IL-18 bioactivity were set-up and evaluated. A specific bioassay was considered as the most reliable test. The stability and integrity of IL-18 was then verified during the encapsulation process. Consequently, two procedures of IL-18 encapsulation in PLGA microparticles (W/O/W and S/O/W) were investigated. As determined by radiolabelling studies using 125I-IL-18 and a continuous flow system, the in vitro release profile of IL-18 was optimum with S/O/W method with a moderate burst effect and a subsequent progressive discharge of 16.5 ± 8.4 ng/day during the next 21 days against 6.1 ± 4.2 ng/day with the W/O/W method. Considering analytical testing of IL-18 together with its preserved biological activity after release from microspheres, amounts of the active cytokine obtained with S/O/W method were relevant to plan in vivo evaluation to validate the therapeutic strategy.
Keywords: Cancer therapy; Adjuvants; Controlled drug delivery systems; Cytokine release; Poly(lactic-co-glycolic acid); Interleukin-18; Microspheres;

Effects of the type of release medium on drug release from PLGA-based microparticles: Experiment and theory by N. Faisant; J. Akiki; F. Siepmann; J.P. Benoit; J. Siepmann (189-197).
The major objectives of the present study were: (i) to prepare 5-fluorouracil (5-FU)-loaded, poly(lactic-co-glycolic acid) (PLGA)-based microparticles, which can be used for the treatment of brain tumors, (ii) to study the effects of the type of release medium on the resulting drug release kinetics, and (iii) to get further insight into the underlying drug release mechanisms. Spherical microparticles were prepared by a solvent extraction method and characterized using different techniques, including size exclusion chromatography (SEC), differential scanning calorimetry (DSC), scanning electron microscopy (SEM) and particle size analysis before and upon exposure to various release media. Interestingly, very different drug release patterns (including mono-, bi- and tri-phasic ones) were observed, depending on the pH, osmolarity and temperature of the release medium. An adequate mathematical theory was used to quantitatively describe the experimentally measured 5-FU release patterns. The model considers the limited solubility of the drug, polymer degradation as well as drug diffusion and allowed to determine system and release medium specific parameters, such as the diffusion coefficient of the drug. In particular, the pH and temperature of the release medium were found to be of major importance for the resulting release patterns. Based on the obtained knowledge the selection of an appropriate release medium for in vitro tests simulating in vivo conditions can be facilitated, and “stress tests” can be developed allowing to get rapid feedback on the release characteristics of a specific batch.
Keywords: Microparticle; Controlled release; PLGA; Release medium; Modeling;

How porosity and size affect the drug release mechanisms from PLGA-based microparticles by D. Klose; F. Siepmann; K. Elkharraz; S. Krenzlin; J. Siepmann (198-206).
Porous, poly(lactic-co-glycolic acid) (PLGA)-based microparticles were prepared using a water-in-oil-in-water (W/O/W) solvent extraction/evaporation technique. Lidocaine was used as a model drug and different-sized particle fractions were obtained by sieving. The physicochemical properties of the devices and changes thereof upon exposure to phosphate buffer pH 7.4 were studied using optical and scanning electron microscopy, size exclusion chromatography (SEC), differential scanning calorimetry (DSC), gravimetric analysis and in vitro drug release measurements. In contrast to non-porous microparticles of identical composition, the relative drug release rate was found to decrease with increasing system size. SEC, DSC and gravimetric analysis showed that the degradation rate of the polymer increased with increasing microparticle dimension, indicating that autocatalytic effects play an important role even in small and highly porous PLGA-based microparticles. However, these effects were much less pronounced than in non-porous devices. Importantly, they were overcompensated by the effects of the increasing diffusion pathway lengths with increasing system dimension. Thus, high initial microparticle porosities do not only lead to increased drug mobilities, but can also fundamentally alter the underlying mass transport mechanisms.
Keywords: Porosity; Autocatalysis; PLGA; Controlled release; Microparticle;

Noticeboard (207-208).