Novel approaches for oral delivery of macromolecules

Traditional forms of administrations of nonabsorbable drugs and peptides often rely on their parenteral injection, since the intestinal epithelium is poorly permeable to these therapeutical agents. A number of innovative drug delivery approaches have been recently developed, including the drug entrapment within small vesicles or their passage through the intestinal paracellular pathway. Zonula occludens toxin, a recently discovered protein elaborated by Vibrio cholerae, provided tools to gain more insights on the pathophysiology of the regulation of intestinal permeability through the paracellular pathway and to develop alternative approaches for the oral delivery of drugs and macromolecules normally not absorbed through the intestine.     

Novel delivery systems for coagulation proteins

Interferon-alpha (IFN-alpha) is an important molecule in the antiviral response, but cells from HIV-1-infected individuals show a reduced ability to secrete IFN-alpha. We investigated an association between an imbalance of type 1/type2 cytokines and the production of IFN-alpha in HIV-1 infection. We used whole blood culture to study the cytokine production profile, interferon-gamma (IFN-gamma) and interleukin-4 (IL-4), in response to HIV-1 antigens and to study the Sendai Virus and HSV-1-induced-production of IFN-alpha in seven HIV-1-infected patients. An impaired synthesis of IFN-alpha was obtained in patients with a predominant IL-4 production (IL-4 > IFN-gamma), and we found a positive correlation between the ex vivo production of IFN-alpha and the IFN-gamma/IL-4 ratio but not with the HIV RNA copy number in plasma. We investigated the role of T-cell-derived cytokines in the in vitro production of IFN-alpha by PBMC from eight healthy donors, activated with Sendai Virus or HSV-1. Whereas type 2 cytokines (IL-4, IL-13) inhibited virus-induced IFN-alpha synthesis, on the contrary, type 1 cytokines (IL-2, IFN-gamma) enhanced it. A disarray in the T-cell-derived cytokine response may play a role in the defect of IFN-alpha production in HIV-1-infected individuals. Further investigations are needed to explore this hypothesis.     

Reversible skin permeabilization for transdermal delivery of macromolecules

Great advances in transdermal delivery of macromolecules have been made within the last few years using ultradeformable liposomes, electroporation, and low-frequency ultrasound, each of which has been shown to deliver macromolecules at clinically useful rates. Until recently, delivery of high-molecular-weight compounds across the skin was not a realistic option, since the skin's great barrier properties prevent transport of macromolecules across human skin at therapeutically relevant rates. An overview of chemical (liposomes and chemical enhancers) and physical (iontophoresis, electroporation, and ultrasound) methods of enhancement is presented, with emphasis on work published within the last two years. Key experimental results and their mechanistic interpretation are provided for each enhancement technique.     

Convection-enhanced delivery of macromolecules in the brain

For many compounds (neurotrophic factors, antibodies, growth factors, genetic vectors, enzymes) slow diffusion in the brain severely limits drug distribution and effect after direct drug administration into brain parenchyma. We investigated convection as a means to enhance the distribution of the large and small molecules 111In-labeled transferrin (111In-Tf; M(r), 80,000) and [14C]sucrose (M(r), 359) over centimeter distances by maintaining a pressure gradient during interstitial infusion into white matter to generate bulk flow through the brain interstitium. The volume of distribution (Vd) containing > or = 1% concentration of infusion solution increased linearly with the infusion volume (Vi) for 111In-Tf(Vd/Vi, 6:1) and [14C]sucrose (Vd/Vi, 13:1). Twenty-four hours after infusion, the distribution of 111In-Tf was increased and more homogeneous, and penetration into gray matter had occurred. By using convection to supplement simple diffusion, enhanced distribution of large and small molecules can be obtained in the brain while achieving drug concentrations orders of magnitude greater than systemic levels.     

Polymeric controlled delivery for immunization

Current vaccine technology has limitations; for example, most vaccines require repeat administration for long-term protection, and immunity at mucosal surfaces is difficult to achieve. In animal models, polymeric controlled-release systems provide long-lasting systemic or mucosal immunoprotection, often after a single administration. Polymeric devices that deliver a controlled amount of antibody can provide passive immunity against genital herpes infections in mice; orally administered polymeric-microsphere-based vaccines produce enhanced immune responses in rodents and primates. These new delivery technologies have many desirable features, and so their use in humans could have a substantial impact on worldwide public health.     

Development of drug delivery systems for macromolecular drugs

With a rapid progress in biotechnology, a variety of endogenous macromolecular substances have become a novel class of therapeutic agents. This review will focus on the development of delivery systems for macromolecular drugs. Current status and future perspectives in this research field are reviewed mainly based on the results obtained in our laboratory. First of all, we studied pharmacokinetic characteristics of macromolecules in relation to their physicochemical properties such as molecular weight and electric charge. Based on this information, we first developed macromolecular prodrugs as a delivery system for low molecular weight drugs. An antitumor antibiotic, mitomycin C (MMC) were covalently conjugated with dextran and various types of macromolecular prodrug of MMC were developed for tumor targeting. Secondly, delivery systems for protein drugs such as soybean trypsin inhibitor, uricase, and recombinant superoxide dismutase (SOD) were developed. In particular, successful targeting of SOD to the liver, kidney and blood circulation was achieved by chemical modification of the protein drug. Finally, we have been trying to develop delivery systems for nucleic acid drugs involving antisense oligonucleotides and plasmid DNA. Prior to the development of delivery systems, we found that the pharmacokinetics of the nucleic acid drugs are decided by their physicochemical properties as polyanions even if these materials contain genetic information. Several approaches were tested to control the in vivo behavior of the oligonucleotides and plasmid DNA based on the finding. Thus, we have established the strategy for rational design of delivery systems for various types of macromolecular drugs based on the pharmacokinetic considerations. This methodology can be a formidable tool for the development of clinically applicable macromolecular drugs.     

In vitro myotoxicity of selected cationic macromolecules used in non-viral gene delivery

PURPOSE: Cationic lipid/DNA complexes have been proposed as a method of in vivo gene delivery via intravenous or intramuscular injection. A concern with using these polycationic molecules is whether they are associated with tissue toxicity at the injection site. Therefore, the objective of these studies was to investigate the myotoxic potential of selected non-viral gene delivery macromolecules (e.g., cationic lipids and polymers) with and without plasmid DNA (pDNA) in vitro. METHODS: Myotoxicity was assessed by the cumulative release of creatine kinase (CK) over 90 minutes from the isolated rodent extensor digitorum longus muscle into a carbogenated balanced salt solution (BBS, pH 7.4, 37 degrees C) following a 15 microL injection of the test formulation. Phenytoin (Dilantin) and normal saline served as positive and negative controls, respectively. RESULTS: The myotoxicity of plasmid DNA (pDNA, approximately 5000bp, 1 mg/ml) was not statistically different from normal saline. However, the myotoxicity of Dilantin was 16-times higher than either normal saline or pDNA (p < 0.05). Cationic liposomes were found to be less myotoxic than polylysine and PAMAM dendrimers. Polylysine's myotoxicity was found to be dependent upon concentration and molecular weight. The myotoxicity of formulations of cationic liposomes(s), lower molecular weight polylysine (25,000) and higher concentration of PAMAM dendrimers with pDNA were found to be statistically less significant than those formulations without pDNA. CONCLUSIONS: The cationic liposomes were less myotoxic compared to the dendrimers and polylysine. Myotoxicity was dependent upon the type of cationic lipid macromolecule, concentration, molecular weight and the presence of pDNA. A possible explanation for this reduced tissue damage in cationic lipids complexed with pDNA is that the formation of complex reduces the overall positive charge of the injectable system resulting in less damage.     

Chitosan microcapsules as controlled release systems for insulin

This study analyses the use of chitosan for the production of surface crosslinked microparticulate systems containing insulin. The microcapsules were prepared by an innovative technique based on interfacial crosslinkage of different amounts of chitosan solubilized in the inner phase of a water/oil emulsion by means of ascorbyl palmitate. The correlation between the main formulation parameters and the functional properties of the microcapsules were analysed. Insulin is incorporated with high efficiency. The peptide release is constant for appreciable periods of time. The content of chitosan modulates the main physico-chemical properties of the matrix.     

Drug delivery systems for cyclosporine: achievements and complications

The review deals with the preparation, properties, and analysis of different kinds of cyclosporine delivery systems, such as solid formulations, liposomes, emulsions and microemulsions and targeted cyclosporine formulations. The review points out a key role of delivery systems in increasing the therapeutic effectiveness of cyclosporine. Comparative studies of the prior marketed formulation, Sandimmune, with a new microemulsion formulation, Neoral, are discussed including some data on clinical development of Neoral.     

Novel high performance,dimensionally controlled PM steels for sinter-hardening

Abstract

Recent trends in the technology, production and applications of diamond tooling are reviewed. The continuing fall in the price of synthetic diamond as production volumes increase provides a strong incentive for tooling companies to reduce the cost of matrix materials and of their manufacturing processes, with an accompanying need for research in these areas. Diamond wires for the cutting of granite and other rock continues to grow in application and provides another focus for materials and technology advances.     

Novel methods of insulin delivery: an update

Conventional insulin therapies involve multiple daily subcutaneous injections. However, the resultant disposition of insulin and blood glucose differs considerably from that following the physiological secretion of pancreatic insulin. A variety of alternative routes/methods have been investigated to improve systemic insulin delivery. Peroral and nasal insulin administration have demonstrated good potential for the treatment of diabetes. Facilitated transdermal delivery has also enjoyed success in promoting the systemic delivery of insulin. In addition, pulmonary, buccal, and ocular insulin administration have been shown to decrease serum glucose concentrations. Other methods that have been investigated for their potential in systemic insulin delivery include rectal, vaginal, and uterine routes.     

Drug delivery systems in gene therapy

The use of nonviral vectors is an attractive in vivo gene delivery strategy that is simpler and lacks some risks inherent in viral systems. Liposomes and receptor-mediated polycation systems are promising carriers for delivery and expression of plasmid DNA encoding genes into the target cells. Many barriers need to be overcome for successful in vivo DNA delivery using these carrier systems. Various factors such as the extent of DNA condensation, particle size of the DNA complex, route of administration, stability against nucleases, target sites, in vivo disposition, binding to cell surface receptor and internalization, intracellular trafficking affect the in vivo gene delivery and expression. This chapter will focus on the current status and perspectives of the plasmid DNA delivery systems for in vivo gene therapy.     

Ocular-specific chemical delivery systems of betaxolol for safe local treatment of glaucoma

This paper examines the social origins of the rise in adult mortality in Russia and selected Eastern European countries. Three explanations for this trend are considered: (1) Soviet health policy, (2) social stress, and (3) health lifestyles. The socialist states were generally characterized by a persistently poor mortality performance as part of a long-term process of deterioration, with particularly negative outcomes for the life expectancy of middle-aged, male manual workers. Soviet-style health policy was ineffective in dealing with the crisis, and stress per se does not seem to be the primary cause of the rise in mortality. Although more research is needed, the suggestion is made that poor health lifestyles--reflected especially in heavy alcohol consumption, and also in smoking, lack of exercise, and high-fat diets--are the major social determinant of the upturn in deaths.