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Current Drug Delivery, Volume 1, No. 1, 2004

 

Contents

 

ATP-Containing Immunoliposomes Specific for Cardiac Myosin Pp.1-7

Wei Liang, Tatyana Levchenko, Ban-An Khaw and Vladimir Torchilin

[Abstract] [Full text article]

 

Biphasic Lipid Vesicles (BiphasixTM) Enhance the Adjuvanticity of CpG Oligonucleotides Following Systemic and Mucosal Administration Pp.9-15

Shawn Babiuk, Maria E. Baca-Estrada, Dorothy M. Middleton, Rolf Hecker, Lorne A. Babiuk  and Marianna Foldvari

[Abstract] [Full text article]

 

Formulation and Characterisation of Tetracycline-Containing Bioadhesive Polymer Networks Designed for the Treatment of Periodontal Disease Pp.17-25

David S. Jones, Michelle S. Lawlor and A. David Woolfson

[Abstract] [Full text article]

 

The Role of ABC Transporters in Drug Resistance, Metabolism and Toxicity Pp.27-42

Hristos Glavinas, Peter Krajcsi, Judit Cserepes  and Balazs Sarkadi

[Abstract] [Full text article]

 

An MDR-EGFP Gene Fusion Allows for Direct Cellular Localization, Function and Stability Assessment of P-Glycoprotein Pp.43-56

Jordi Petriz, Michael M. Gottesman  and Josep M. Aran

[Abstract] [Full text article]

 

Pain Control Via Opioid Analgesic- Local Anesthetic Loaded IPNs Pp.57-64

Dilek Sendil Keskin, Donald L.Wise  and Vasif Hasirci

[Abstract] [Full text article]

 

Development of Topically Effective Formulations of Acetazolamide Using HP-β-CD-Polymer Co-Complexes Pp.65-72

Indu Pal Kaur, Mona Kapil, R. Smitha and Deepika Aggarwal

[Abstract] [Full text article]

 

Challenges in Current Drug Delivery from the Potential Application of Pharmacogenomics and Personalized Medicine in Clinical Practice Pp.73-80

Ioannis S. Vizirianakis

[Abstract] [Full text article]

 

Physical Enhancement of Transdermal Drug Application: Is Delivery Technology Keeping up with Pharmaceutical Development? Pp.81-92

S.E. Cross and M.S. Roberts

[Abstract] [Full text article]

 

Abstracts

 

[Back to top] ATP-Containing Immunoliposomes Specific for Cardiac Myosin

Wei Liang, Tatyana Levchenko, Ban-An Khaw and Vladimir Torchilin
[Full text article]

 

The application of ATP-loaded liposomes has been shown effective against ischemic damage in several tissues. In this study, we have prepared ATP-containing liposomes capable of specific recognition of component (myosin) specific for ischemic myocardium. ATP-containing immunoliposomes specific towards cardiac myosin were obtained by the attachment of the monoclonal anti-cardiac myosin 2G4 antibody to the surface of ATP-containing PEGylated liposomes prepared by the freezing-thawing method. Since intracellular myosin is exposed only in the areas containing ischemically compromised cells with damaged plasmic membranes, such liposomes are expected to target these areas both in vitro and in vivo. The attachment of the antibody did not provoke their ATP release from the liposomes and only minimally influenced liposome size and size distribution. Liposome-attached anti-myosin 2G4 antibody preserved its specific activity; and anti-myosin antibody-bearing, ATP-loaded liposomes bound efficiently to the monolayer of myosin in ELISA. The preparation of myosin-specific ATP-loaded immunoliposomes represented an important step in the development of targeted delivery systems capable of providing energy support to ischemic myocardium in vivo.

 

[Back to top] Biphasic Lipid Vesicles (BiphasixTM) Enhance the Adjuvanticity of CpG Oligonucleotides Following Systemic and Mucosal  Administration

Shawn Babiuk, Maria E. Baca-Estrada, Dorothy M. Middleton, Rolf Hecker, Lorne A. Babiuk  and Marianna Foldvari

[Full text article]

 

CpG oligonucleotides (ODNs) are potent mucosal and systemic adjuvants. For practical applications however, improvements in delivery need to be developed. A mouse model was used to determine if the biological activity of CpG ODNs could be enhanced using a novel delivery system of biphasic lipid vesicles (BiphasixTM Vaccine-Targeting Adjuvant; VTA).

 

Immunization studies were performed to evaluate the potential of VTA formulations to enhance the immunoadjuvant activity of CpG ODNs following systemic or mucosal administration with gD. Immune responses following immunization were assessed by protection from HSV-1 viral challenge and characterization of serum gD-specific antibody responses using ELISA.

 

VTA formulations in combination with CpG and glycoprotein D (gD) were able to increase gD-specific IgG in serum compared to gD alone, and protect from a lethal HSV-1 challenge following subcutaneous immunization. Following mucosal immunization, VTA formulations in combination with CpG and antigen enhanced mucosal IgA responses compared to CpG and antigen administered in PBS.

 

[Back to top] Formulation and Characterisation of Tetracycline-Containing Bioadhesive Polymer Networks Designed for the Treatment of Periodontal Disease

David S. Jones, Michelle S. Lawlor and A. David Woolfson

[Full text article]

 

This study described the drug release, rheological (dynamic and flow) and textural/mechanical properties of a series of formulations composed of 15% w/w polymethylvinylether-co-maleic anhydride (PMVE-MA), 0–9% w/w polyvinylpyrrolidone (PVP) and containing 1–5% w/w tetracycline hydrochloride, designed for the treatment of periodontal disease. All formulations exhibited pseudoplastic flow with minimal thixotropy. Increasing the concentration of PVP sequentially increased the zero-rate viscosity (derived from the Cross model) and the hardness and compressibility of the formulations (derived from texture profile analysis). These affects may be accredited to increased polymer entanglement and, in light of the observed synergy between the two polymers with respect to their textural and rheological properties, interaction between PVP and PMVEMA. Increasing the concentration of PVP increased the storage and loss moduli yet decreased the loss tangent of all formulations, indicative of increased elastic behaviour. Synergy between the two polymers with respect to their viscoelastic properties was observed. Increased adhesiveness, associated with increased concentrations of PVP was ascribed to the increasing bioadhesion and tack of the formulations. The effect of increasing drug concentration on therheological and textural properties was dependent on PVP concentration. At lower concentrations (0, 3% w/w) no effect was observed whereas, in the presence of 9% w/w PVP, increasing drug concentration increased formulation elasticity, zero rate viscosity, hardness and compressibility. These observations were ascribed to the greater mass of suspended drug in formulations containing the highest concentration of PVP. Drug release from formulations containing 6 and 9% PVP (and 5% w/w drug) was prolonged and swelling/diffusion controlled. Based on the drug release, rheological and textural properties, it is suggested that the formulation containing 15% w/w PMVE-MA, 6% w/w PVP and tetracycline hydrochloride (5% w/w) may be useful for the treatment of periodontal disease.

 

[Back to top] The Role of ABC Transporters in Drug Resistance, Metabolism and Toxicity

Hristos Glavinas, Peter Krajcsi, Judit Cserepes  and Balazs Sarkadi

[Full text article]

 

ATP Binding Cassette (ABC) transporters form a special family of membrane proteins, characterized by homologous ATP-binding, and large, multispanning transmembrane domains. Several members of this family are primary active transporters, which significantly modulate the absorption, metabolism, cellular effectivity and toxicity of pharmacological agents. This review provides a general overview of the human ABC transporters, their expression, localization and basic mechanism of action. Then we shortly deal with the human ABC transporters as targets of therapeutic interventions in medicine, including cancer drug resistance, lipid and other metabolic disorders, and even gene therapy applications. We place a special emphasis on the three major groups of ABC transporters involved in cancer multidrug resistance (MDR). These are the classical P-glycoprotein (MDR1, ABCB1), the multidrug resistance associated proteins (MRPs, in the ABCC subfamily), and the ABCG2 protein, an ABC half-transporter. All these proteins catalyze an ATP-dependent active transport of chemically unrelated compounds, including anticancer drugs. MDR1 (P-glycoprotein) and ABCG2 preferentially extrude large hydrophobic, positively charged molecules, while the members of the MRP family can extrude both hydrophobic uncharged molecules and water-soluble anionic compounds. Based on the physiological expression and role of these transporters, we provide examples for their role in Absorption-Distribution- Metabolism-Excretion (ADME) and toxicology, and describe several basic assays which can be applied for screening drug interactions with ABC transporters in the course of drug research and development.

 

[Back to top] An MDR-EGFP Gene Fusion Allows for Direct Cellular Localization, Function and Stability Assessment of P-Glycoprotein

Jordi Petriz, Michael M. Gottesman  and Josep M. Aran

[Full text article]

 

In cancer and AIDS, overexpression of the MDR1 gene has important implications in the design of chemotherapy protocols because of the ability of its product, the ATPdependent drug efflux pump P-glycoprotein (Pgp), to confer selective advantage to tumor and HIV-infected cells in the form of multidrug resistance. To study Pgp expression and physiology, we designed a translational fusion between the MDR1 and enhanced green fluorescent protein (EGFP) genes. The chimeric protein, Pgp-EGFP, was concentrated mainly in the plasma membrane and in the Golgi when expressed in drug-sensitive KB- 3-1 cells. Doxorubicin, daunorubicin and rhodamine-123 efflux assays confirmed function of the chimeric pump. Also, at the single-cell level, an inverse relationship between Pgp-EGFP expression and nuclear doxorubicin accumulation was demonstrated. Polarized Pgp expression on the apical cell surface was confirmed by transfection of the MDR-EGFP fusion gene into MDCK cells. However, after colchicine selection, Pgp-EGFP was also detectable in the lateral domain of the transfected MDCK monolayers. These results indicate that drug selection affects not only expression, but cellular localization of Pgp. Furthermore, using a tet-based inducible expression system for Pgp-EGFP, we confirmed the stable nature of Pgp (t1/2 total Pgp-EGFP= 2.2 days), but revealed that surface-Pgp acquires extra stability as an active pump (t1/2 surface Pgp-EGFP= 3.7 days).

 

[Back to top] Pain Control Via Opioid Analgesic- Local Anesthetic Loaded IPNs

Dilek Sendil Keskin, Donald L.Wise  and Vasif Hasirci

[Full text article]

 

Relief of chronic pain is an important clinical problem requiring special care and approaches. The present study was designed for the construction of a controlled release system for local application of analgesics (hydromorphone (HM), morphine (M), and codeine (C)) and a local anesthetic, bupivacaine (BP). An interpenetrating network (IPN) drug release system was prepared by using a biocompatible, biodegradable copolyester, poly(3-hydroxybutyrate-co-3- hydroxyvalerate) (PHBV) and another biocompatible but synthetic, nondegradable polymer, poly (2- hydroxyethyl methacrylate), (PHEMA). In situ release kinetics of the IPN system was first order for BP but could not be fitted to any known equation for the other drugs. Complete release from the IPNs occurred within a considerably short time (24 h for 80 % of the drugs) most probably due to the significant hydrophilicity of PHEMA. In order to slow down the release rate these IPNs were coated with PHBV. Release from these coated IPNs (cIPN) resulted in rates that could be described by Higuchi’s equations. In vivo measurement of antinociceptive efficacy was carried out in rats with tail flick and paw-withdrawal tests after inducing chronic pain created by sciatic nerve ligation at the right side. Control groups received placebo implants. In vivo studies showed potent, prolonged (2-3 days) antinociception at the site of injury (right paw) for strong opioids (HM and M) and about 2 days for the weak opioid (C) and local anesthetic (BP). In all cases the release rate was found to be as important as the antinociceptive potency. The weakest opioid analgesic of those evaluated (C) had a higher first day antinociception than its stronger counterpart M, probably due to its higher initial concentration that was expected from its faster release rate in the in situ experiments.

 

[Back to top] Development of Topically Effective Formulations of Acetazolamide Using HP-β-CD-Polymer Co-Complexes

Indu Pal Kaur, Mona Kapil, R. Smitha and Deepika Aggarwal

[Full text article]

 

The myriad of side effects induced by acetazolamide (oral use), the introduction of newer topical carbonic anhydrase inhibitors (CAI’s) and the advent of other antiglaucoma medications has led to a decreased interest in acetazolamide. The use of cyclodextrins to improve the solubility and bioavailability of poorly soluble drugs has however, rekindled an interest in acetazolamide (ACZ), because its poor solubility is one of the major factor responsible for its failure to show topical effectiveness. Since water soluble polymers have been reported to improve the complexing capabilities of b-cyclodextrins, in the present study water soluble polymers like polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), hydroxypropymethylcellulose (HPMC) and the mucoadhesive polymer Carbopol 934P were incorporated into aqueous 10% w/v 2HP-b-CD solution to improve the solubility of ACZ. The effect of these polymers on the corneal transport of 5 mg/ml (0.5% w/v) solution of ACZ in aqueous 10% 2HP-b-CD was evaluated. The inclusion of these polymers significantly increased the solubility of ACZ from 3.43 mg/ml in aqueous 10% 2HP-b-CD to 5.1 mg/ml (48.6%) in 0.05% PVP; 6.80 mg/ml (98.3%) in 0.05% PVA; and 6.74 mg/ml (96.5%) in 0.2% Carbopol 934P. From amongst the various polymers assessed in the study, PVA was deemed the best, based on the premise of better apparent permeability coefficient (Papp) upon in vitro corneal permeation studies. Inspite of a large enhancement in solubility produced by Carbopol 934P, surprisingly, it could not efficiently increase the Papp.

 

[Back to top] Challenges in Current Drug Delivery from the Potential Application of Pharmacogenomics and Personalized Medicine in Clinical Practice

Ioannis S. Vizirianakis

[Full text article]

 

The recent technological achievements in biotechnology and recombinant DNA technology have provided multiple new methods, molecular targets, and DNA-based diagnostics to pharmaceutical research that can be utilized in assays for screening and developing potential biotechnology-based drugs, as well as in biomedicine, health and pharmaceutical care. Furthermore, such advances opened up new opportunities by applying genetic information data in pharmacotherapy and drug delivery, thus ensuring better drug efficacy and safety in clinical practice. Now the concepts of personalized medicine and pharmacogenomics are likely improving the area of pharmacodynamics and pharmacokinetics, since they favor differentiation of the conventional clinical diagnosis and drug selection into separate molecular subtypes of individuals belonging within a group of patients suffering from the same disease. Genetic polymorphisms have already been detected and analyzed in genes encoding drug-metabolizing enzymes, transporters as well as targets (e.g. receptors). The potential of applying genotyping and haplotyping analysis in future pharmaceutical care could eventually lead to pharmacotyping, i.e. individualized drug delivery profiling based on genetic-bioinformatic data in routine patient care. However, the steps towards this direction of drug delivery in clinical practice still have a long way to go to be fully achieved; until then, the critical evaluation of all available clinical data including pharmacodynamic, pharmacokinetic and genomic must be assessed for ensuring drug efficacy and safety. In this way, there has been great progress in elucidating genetic determinants contributing to the observed interindividual differences in drug disposition and effects, thus implementing current drug delivery with molecular genetics and diagnostics.

 

[Back to top] Physical Enhancement of Transdermal Drug Application: Is Delivery Technology Keeping up with Pharmaceutical Development?

S.E. Cross and M.S. Roberts

[Full text article]

 

Advances in molecular biology have given us a wide range of protein and peptide-based drugs that are unsuitable for oral delivery because of their high degree of first-pass metabolism. Though parenteral delivery is the obvious answer, for the successful development of commercial chronic and self-administration usage formulations it is not the ideal choice. Transdermal delivery is emerging as the biggest application target for these agents, however, the skin is extremely efficient at keeping out such large molecular weight compounds and therapeutic levels are never going to be realistically achieved by passive absorption. Physical enhancement mechanisms including: iontophoresis, electroporation, ultrasound, photomechanical waves, microneedles and jet-propelled particles are emerging as solutions to this topical delivery dilemma. Adding proteins and peptides to the list of other large molecular weight drugs with insufficient passive transdermal fluxes to be therapeutically useful, we have a collection of pharmacological agents waiting for efficient delivery methods to be introduced. This article reviews the current state of physical transdermal delivery technology, assesses the pros and cons of each technique and summarises the evidence-base of their drug delivery capabilities.