Co-formulations of adalimumab with hyaluronic acid/polyvinylpyrrolidone to combine intraarticular drug delivery and viscosupplementation.

Polymer-based formulations present an attractive strategy in intraarticular drug-delivery to refrain biologicals from early leakage from the joint. In this study, co-formulations of hyaluronic acid and polyvinylpyrrolidone were investigated for their potential as viscosupplements and their influence on the transsynovial loss of adalimumab. For this purpose, polymer mixtures were evaluated for their viscosity and elasticity behavior while their influence on the permeation of adalimumab across a porcine ex-vivo synovial membrane was determined. Hyaluronic acid showed strong shear thinning behavior and exhibited high viscosity and elasticity at low motions, while combinations with polyvinylpyrrolidone provided absorption and stiffness at high mechanical stress, so that they can potentially restore the rheological properties of the synovial fluid over the range of joint motion. In addition, the formulations showed significant influence on transsynovial permeation kinetics of adalimumab and hyaluronic acid, which could be decelerated up to 5- and 3-fold, respectively. Besides viscosity effects, adalimumab was retained primarily by an electrostatic interaction with hyaluronic acid, as detected by isothermal calibration calorimetry. Furthermore, polymer-mediated stabilization of the antibody activity was detected. In summary, hyaluronic acid - polyvinylpyrrolidone combinations can be efficiently used to prolong the residence of adalimumab in the joint cavity while simultaneously supplying viscosupplementation.

Hyaluronic Acid Increases Anti-Inflammatory Efficacy of Rectal 5-Amino Salicylic Acid Administration in a Murine Colitis Model.

5-amino salicylic acid (5-ASA) is a standard therapy for the treatment of mild to moderate forms of inflammatory bowel diseases (IBD) whereas more severe forms involve the use of steroids and immunosuppressive drugs. Hyaluronic acid (HA) is a naturally occurring non-sulfated glycosaminoglycan that has shown epithelium protective effects in experimental colitis recently. In this study, both 5-ASA (30 mg/kg) and HA (15 mg/kg or 30 mg/kg) were administered rectally and investigated for their potential complementary therapeutic effects in moderate or severe murine colitis models. Intrarectal treatment of moderate and severe colitis with 5-ASA alone or HA alone at a dose of 30 mg/kg led to a significant decrease in clinical activity and histology scores, myeloperoxidase activity (MPO), TNF-α, IL-6 and IL-1ß in colitis mice compared to untreated animals. The combination of HA (30 mg/kg) and 5-ASA in severe colitis led to a significant improvement of colitis compared to 5-ASA alone. Combined rectal therapy with HA and 5-ASA could be a treatment alternative for severe cases of IBD as it was the only treatment tested that was not significantly different from the healthy control group. This study further underlines the benefit of searching for yet unexplored drug combinations that show therapeutic potential in IBD without the need of designing completely new drug entities.

Anti-inflammatory effects of acacia and guar gum in 5-amino salicylic acid formulations in experimental colitis.

Findings from recent studies revealed a significant anti-inflammatory effect of polysaccharide-based excipients when formulated with classical drugs in experimental inflammatory bowel disease models. In this study, acacia and guar gum were investigated beyond their typical functionality for a possible additive anti-inflammatory effect when administered with 5-amino salicylic acid (5ASA) in murine experimental colitis. Anti-inflammatory effects of acacia and guar gum-based aqueous suspensions of 5ASA were evaluated in a murine experimental colitis. Acacia or guar gum (30 or 300 mg/kg) were administered via rectal administration alone or in combination with 5ASA (30 mg/kg). Disease activity, myeloperoxidase activity (MPO) and intratissue concentrations of various cytokines were assessed. Both acacia and guar gum separately showed significant effects in reducing the inflammatory markers in murine colitis model in vivo. When combined with the anti-inflammatory drug 5ASA, acacia showed a stronger therapeutic effect than guar gum, especially at the higher dose of acacia (300 mg/kg) which significantly reduced the inflammation in vivo compared to 5ASA alone (MPO, 5ASA: 5743 ± 1334, 5ASA + 30 mg/kg acacia: 3762 ± 2342; 5ASA + 30 mg/kg guar gum: 7373 ± 2115, 5ASA + 300 mg/kg acacia: 3131 ± 1012, 5ASA + 300 mg/kg guar gum: 6358 ± 2379; all U/g tissue). Acacia and guar gum separately showed significant anti-inflammatory effects in murine colitis, and furthermore, high dose acacia led to an additional therapeutic benefit when co-administered with 5ASA. These results indicate that further investigations are surely warranted in the search of better colitis therapy.

An ex-vivo model for transsynovial drug permeation of intraarticular injectables in naive and arthritic synovium.

Estimation of joint residence time of a drug is a key requirement for rational development of intraarticular therapeutics. There is a great need for a predictive model to reduce the high number of animal experiments in early stage development. Here, a Franz-cell based porcine ex-vivo permeation model is proposed, and transsynovial permeation of fluorescently-labeled dextrans in the range of potential drug candidates (10-150 kDa), as well as a small molecule (fluorescein sodium) and charged dextran derivates, have been determined. In addition, a lipopolysaccharide (LPS) -induced synovitis model was assessed for inflammatory biomarker levels and its effect on permeation of the solutes. Size-dependent permeability was observed for the analytes, which distinctly differed from findings with an artificial polycarbonate membrane, which is a widely used model. LPS was found to successfully stimulate an inflammatory response and led to a reduced size selectivity of the synovial membrane. 150 kDa dextran flux was accelerated approximately 2.5-fold in the inflamed state, whereas the permeation of smaller molecules was little affected. Moreover, by varying the LPS concentrations, the ex-vivo model was shown to produce varying degrees of synovitis-like inflammation. A simple and highly relevant ex-vivo tool for investigation of transsynovial permeation was developed, offering the further advantage of mimicking synovitis-induced permeability changes. Thus, this model provides a promising method for formulation screening, while reducing the need for animal experiments.

Anti-Inflammatory Activity of Chitosan and 5-Amino Salicylic Acid Combinations in Experimental Colitis.

Chitosan is used in various drug delivery approaches as a pharmaceutical excipient. Although its potential as an immunomodulatory agent has been reported, its use in this capacity has not been fully explored. The efficacy of chitosan as an active pharmacological agent, particularly in anti-inflammatory therapy in inflammatory bowel diseases (IBD), was investigated in this study. The potential impact of the molecular weight (MW) and degree of deacetylation (DD) of chitosan was investigated together with 5-amino salicylic acid (5-ASA) for its efficacy in a combination anti-inflammatory therapy in murine experimental colitis. Such a combination would potentially be developed into novel dual strategies whereby chitosan acts as a mucoadhesive excipient as well as provide an additional anti-inflammatory benefit. Chitosan grades with different MW and DD were administered intrarectally alone or in combination with 5-ASA to colitis mice for 3 days. Myeloperoxidase (MPO) and alkaline phosphatase (ALP) activity and tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and nuclear factor kappa-B (NF-κB) levels were assessed from the colon. Intrarectal treatment of colitis with 30 mg/kg chitosan alone and with 30 mg/kg 5-ASA for 3 days led to a significant decrease in MPO, ALP, TNF-α, IL-6, IL-1ß and NF-κB in colitis mice compared to untreated mice. Surprisingly, the efficacy of chitosan as an anti-inflammatory polymer was relatively independent from its structural properties, namely DD and MW. However, combinations of chitosan with 5-ASA showed a significant pharmacological improvement, whereby the additive anti-inflammatory efficacy observed shows the possibility of finetuning chitosan by combining it with anti-inflammatory agents to optimize its anti-inflammatory potential.

Viscosupplementation may preserve tibial cartilage and collagen in osteoarthritis: findings from a preclinical model of osteoarthritis.

PURPOSE: Intraarticular (IA) hyaluronic acid (HA) injection is used to reduce pain and improve mobility in knee osteoarthritis (OA). Little is known about histopathological changes underlying HA efficacy. This study investigated dose-related effects of 1% sodium hyaluronate (BioHA) on knee joint histopathology and pain responses in a medial meniscal tear (MMT) rat model of OA. METHODS: Following MMT surgery, rats were randomized into treatment groups: single IA injection of vehicle, BioHA, or an avian-derived hyaluronic acid (hylan G-F 20) on Day 7; or 3 weekly injections of vehicle or BioHA on Days 7, 14, and 21. On Day 35, joints were evaluated by microscopic histopathology for cartilage degeneration, collagen degeneration, synovitis, and cytokine expression (tumor necrosis factor α, transforming growth factor ß). RESULTS: Joint pathology for control animals was consistent with that expected for the MMT model. Rats treated with 3 injections of IA-BioHA had significantly reduced collagen degeneration (21%) relative to control animals. No significant change in collagen degeneration was observed for rats given a single injection of hylan G-F 20 or IA-BioHA compared to control animals. HA treatment did not affect cytokine expression. CONCLUSIONS: IA-BioHA viscosupplementation in a rat MMT model of OA showed preservation of joint cartilage and collagen. This effect was most pronounced on tibial surfaces having less severe injury, suggesting that treatment should be initiated early in the disease process. A comparison of responses to IA-BioHA or hylan G-F 20 in the MMT rat OA model suggest IA-BioHA may be more effective in preserving joint connective tissue.

Stir-bar sorptive extraction and thermal desorption-ion mobility spectrometry for the determination of trinitrotoluene and l,3,5-trinitro-l,3,5-triazine in water samples.

Stir-bar sorptive extraction (SBSE) is interfaced to ion mobility spectrometry (IMS) for the rapid detection of trace analytes, with the explosives, trinitrotoluene (TNT) and l,3,5-trinitro-l,3,5-triazine (RDX) shown as examples. SBSE retains its inherent advantages as a sensitive, straightforward, solventless, and inexpensive method. Additionally, the new SBSE-IMS technique exhibits excellent sensitivity, has onsite field analysis capabilities and provides the potential to detect and quantitate analytes that are difficult to accomplish using gas chromatography (GC) or high-performance liquid chromatography (HPLC). The SBSE-IMS technique is shown to be an effective method for the low-level detection of TNT and RDX from water with method standard deviation of 8.6% for TNT and 6.6% for RDX. The short desorption time of 60 s and analysis time of less than 20 ms along with limits of detection of 0.1 ng/mL for TNT and 1.5 ng/mL for RDX and render the method potentially useful for trace analysis. Desorption profiles showing the kinetics of analyte transfer from the stir-bar into the IMS are shown and discussed; the SBSE-IMS configuration shows very rapid desorption from the stir-bar, with the analytes completely transferred in most cases, in under 1 min.

Determination of parabens in pharmaceutical formulations by solid-phase microextraction-ion mobility spectrometry.

Solid-phase microextraction (SPME) coupled with ion mobility spectrometry (IMS) was used for the detection and quantitation of 4-hydroxybenzoate preservatives, methylparaben, ethylparaben, propylparaben, and butylparaben, in commercial pharmaceutical products. For the first time, SPME-IMS is described for the simultaneous detection, separation, and quantitation of multiple analytes in complex matrixes. The parabens are extracted from the samples using SPME, and the analytes on the fiber are heated by the IMS desorber unit and vaporized into the drift tube. The four preservatives differing only by a methyl group were separated in less than 18 ms. The analytical procedure was optimized for fiber coating selection, extraction time, sample pH, sample volume, ionic strength, and IMS conditions. Separation characteristics such as resolution, theoretical plates, and drift times of the parabens were also evaluated based on the direct interfacing of SPME to IMS. The conditions were tested using six over-the-counter topical products containing various combinations of preservatives. Analysis of the samples by SPME-IMS using benzyl paraben as an internal standard yields good comparison to an HPLC method, thereby reinforcing the applicability of this technique as a method for routine analysis. Limits of detection were 10 ng/mL for methylparaben and ethylparaben and 5 ng/mL for propylparaben and butylparaben. Good linearity range and reproducibility of less than 8% were obtained.

Solid phase micro-extraction coupled with ion mobility spectrometry for the analysis of ephedrine in urine.

Quantitative solid phase micro-extraction (SPME) coupled with ion mobility spectrometry is demonstrated using the analysis of ephedrine in urine. Since its inception in the 1970's ion mobility spectrometry (IMS) has evolved into a useful technique for laboratories to detect explosives, chemical warfare agents, environment pollutants and, increasingly, for detecting drugs of abuse. Ephedrine is extracted directly from urine samples using SPME and the analyte on the fiber is heated by the IMS desorber unit and vaporized into the drift tube. The analytical procedure was optimized for fiber coating selection, extraction temperature, extraction time, sample pH, and analyte desorption temperature. The carryover effects, ion fragmentation characteristics, peak shapes, and drift times of ephedrine were also evaluated based on the direct interfacing of SPME to IMS. A limit of detection of 50 ng/mL of ephedrine in urine and a linear range of 3 orders of magnitude were obtained, showing that SPME-IMS compares well to other techniques for ephedrine and drug analysis presented in the literature.