PEGylated Liposomal Methyl Prednisolone Succinate does not Induce Infusion Reactions in Patients: A Correlation Between in Vitro Immunological and in Vivo Clinical Studies.

PEGylated nanomedicines are known to induce infusion reactions (IRs) that in some cases can be life-threatening. Herein, we report a case study in which a patient with rare mediastinal and intracardiac IgG4-related sclerosing disease received 8 treatments of intravenously administered PEGylated liposomal methylprednisolone-succinate (NSSL-MPS). Due to the ethical requirements to reduce IRs, the patient received a cocktail of premedication including low dose of steroids, acetaminophen and H2 blockers before each infusion. The treatment was well-tolerated in that IRs, complement activation, anti-PEG antibodies and accelerated blood clearance of the PEGylated drug were not detected. Prior to the clinical study, an in vitro panel of assays utilizing blood of healthy donors was used to determine the potential of a PEGylated drug to activate complement system, elicit pro-inflammatory cytokines, damage erythrocytes and affect various components of the blood coagulation system. The overall findings of the in vitro panel were negative and correlated with the results observed in the clinical phase.

Effect of Treating Acute Optic Neuritis With Bioequivalent Oral vs Intravenous Corticosteroids: A Randomized Clinical Trial.

Importance: Intravenous (IV) administration of corticosteroids is the standard of care in the treatment of acute optic neuritis. However, it is uncertain whether a bioequivalent dose of corticosteroid administered orally, which may be more cost-efficient and convenient for patients, is as effective as IV administration in the treatment of acute optic neuritis. Objective: To determine whether recovery of vision following treatment of acute optic neuritis with a high-dose IV corticosteroid is superior to that with a bioequivalent dose of an oral corticosteroid. Design, Setting, and Participants: This single-blind (participants unblinded) randomized clinical trial with 6-month follow-up was conducted at a single tertiary care center in London, Ontario, Canada. Participants were enrolled from March 2012 to May 2015, with the last participant's final visit occurring November 2015. Patients 18 to 64 years of age presenting within 14 days of acute optic neuritis onset, without any recovery at time of randomization and without history of optic neuritis in the same eye, were screened. Inclusion criteria included best-corrected visual acuity (BCVA) of 20/40 or worse and corticosteroids deemed required by treating physician. In total, 89 participants were screened; 64 were eligible, but 9 declined to participate. Thus, 55 participants were enrolled and randomized. Primary analysis was unadjusted and according to the intention-to-treat principle. Interventions: Participants were randomized 1:1 to the IV methylprednisolone sodium succinate (1000-mg) or oral prednisone (1250-mg) group. Main Outcomes and Measures: Primary outcome was recovery of the latency of the P100 component of the visual evoked potential at 6 months. Secondary outcomes were the P100 latency at 1 month and BCVA as assessed with Early Treatment Diabetic Retinopathy Study letter scores on the alphabet chart and scores on low-contrast letters at 1 and 6 months. Results: Of 55 randomized participants, the final analyzed cohort comprised 23 participants in the IV and 22 in the oral treatment groups. The mean (SD) age of the cohort was 34.6 (9.5) years, and there were 28 women (62.2%). At 6 months' recovery, P100 latency in the IV group improved by 62.9 milliseconds (from a mean [SD] of 181.9 [53.6] to 119.0 [16.5] milliseconds), and the oral group improved by 66.7 milliseconds (from a mean [SD] of 200.5 [67.2] to 133.8 [31.5] milliseconds), with no significant difference between groups (P = .07). Similarly, no significant group difference was found in the mean P100 latency recovery at 1 month. For BCVA, recovery between the groups did not reach statistical significance at 1 month or 6 months. In addition, improvements in low-contrast (1.25% and 2.5%) BCVA were not significantly different between treatment groups at 1 or 6 months' recovery. Conclusions and Relevance: This study finds that bioequivalent doses of oral corticosteroids may be used as an alternative to IV corticosteroids to treat acute optic neuritis. Trial Registration: clinicaltrials.gov Identifier: NCT01524250.

Continuous infusion of methylprednisolone via paediatric parenteral nutrition: a pharmacokinetic animal study.

BACKGROUND & AIMS: The aim of our study consisted to measure the pharmacokinetic parameters of methylprednisolone administered in a continuous infusion of a paediatric parenteral nutrition mixture for 24h in the rabbit. METHODS: Fourteen rabbits were split into two groups and assigned a different administration vehicle (all-in-one or two-in-one nutrition mixture). We used USC PACK* pharmacokinetics software to compare the influence of the composition of the paediatric parenteral nutritional solutions on the values of the pharmacokinetic parameters of methylprednisolone. RESULTS: Neither the steady-state plasma concentrations of methylprednisolone hemisuccinate nor the values of the pharmacokinetic parameters of methylprednisolone differed significantly when administered in two-in-one or all-in-one nutrition mixtures. CONCLUSIONS: The composition of the nutritional medium had no discernable effect on the bioavailability of methylprednisolone. Neither the speed at which the steady-state plasma concentration was reached, nor the values of the pharmacokinetic parameters of methylprednisolone were significantly modified.

Nano-Drugs Based on Nano Sterically Stabilized Liposomes for the Treatment of Inflammatory Neurodegenerative Diseases.

The present study shows the advantages of liposome-based nano-drugs as a novel strategy of delivering active pharmaceutical ingredients for treatment of neurodegenerative diseases that involve neuroinflammation. We used the most common animal model for multiple sclerosis (MS), mice experimental autoimmune encephalomyelitis (EAE). The main challenges to overcome are the drugs' unfavorable pharmacokinetics and biodistribution, which result in inadequate therapeutic efficacy and in drug toxicity (due to high and repeated dosage). We designed two different liposomal nano-drugs, i.e., nano sterically stabilized liposomes (NSSL), remote loaded with: (a) a "water-soluble" amphipathic weak acid glucocorticosteroid prodrug, methylprednisolone hemisuccinate (MPS) or (b) the amphipathic weak base nitroxide, Tempamine (TMN). For the NSSL-MPS we also compared the effect of passive targeting alone and of active targeting based on short peptide fragments of ApoE or of ß-amyloid. Our results clearly show that for NSSL-MPS, active targeting is not superior to passive targeting. For the NSSL-MPS and the NSSL-TMN it was demonstrated that these nano-drugs ameliorate the clinical signs and the pathology of EAE. We have further investigated the MPS nano-drug's therapeutic efficacy and its mechanism of action in both the acute and the adoptive transfer EAE models, as well as optimizing the perfomance of the TMN nano-drug. The highly efficacious anti-inflammatory therapeutic feature of these two nano-drugs meets the criteria of disease-modifying drugs and supports further development and evaluation of these nano-drugs as potential therapeutic agents for diseases with an inflammatory component.

Ketoconazole effects on methylprednisolone disposition and their joint suppression of endogenous cortisol.

The effects of ketoconazole and methylprednisolone on endogenous cortisol were studied in eight normal subjects. Intravenous methylprednisolone sodium succinate was given alone in doses of 15 or 30 mg. The methylprednisolone dose was reduced by 57% when ketoconazole was administered chronically for 1 week to seek equivalent methylprednisolone AUCs by compensating for the expected reduction in methylprednisolone clearance. Ketoconazole decreased clearance by 46% and increased mean residence time by 37%. The ratio of the cortisol AUC during each drug treatment compared with baseline conditions was used to assess the net extent and duration of cortisol suppression. This cortisol AUC ratio was reduced from 0.45 (methylprednisolone) to 0.39 (methylprednisolone plus ketoconazole), suggesting that ketoconazole modestly enhanced (P less than 0.01) cortisol suppression. Based on the reduction in methylprednisolone clearance and cortisol AUC by ketoconazole, a 50% lower dose of methylprednisolone during concomitant therapy with ketoconazole is recommended.

Pharmacokinetics and pharmacodynamics of methylprednisolone in obesity.

Methylprednisolone pharmacokinetics and its directly suppressive effects on plasma cortisol, blood histamine (basophils), and circulating helper T cells were evaluated in six obese (at least 35% above ideal body weight) men and six nonobese male volunteers. Methylprednisolone doses of 0.6 mg/kg total body weight were administered as the 21-succinate sodium salt. Absolute clearance (in liters per hour) of methylprednisolone was 40% less in the obese subjects. Total volume of distribution (Vss) of methylprednisolone was unchanged (about 120 L), but when normalized for total body weight, Vss per kilogram was less in obesity. The patterns of cortisol, blood histamine, and helper T cell responses after methylprednisolone administration were similar in both groups, but more profound effects were observed in the obese subjects. Pharmacodynamic models were applied for these immediate effects of methylprednisolone based on the premise that receptor interactions of steroids are followed by rapid suppression of the circadian rhythm of cortisol and recirculation of basophils and helper T cells, which persist until inhibitory concentrations (IC50) of methylprednisolone disappear. Similar IC50 values for the three effects were obtained in both groups, indicating no intrinsic pharmacodynamic differences in sensitivity to these methylprednisolone effects in obesity. However, methylprednisolone should be administered on the basis of ideal body weight, and the dosing interval should be potentially lengthened because of decreased methylprednisolone clearance in obesity.

Pharmacokinetics and pharmacodynamic modeling of direct suppression effects of methylprednisolone on serum cortisol and blood histamine in human subjects.

Pharmacodynamic models for "directly suppressive" effects of methylprednisolone are based on the premise that receptor interactions of steroids are followed by immediate suppression of either the circadian secretion of cortisol or the constant rate recirculation of histamine-containing basophils that persists until inhibitory concentrations of methylprednisolone disappear. Methylprednisolone doses of 0, 10, 20, and 40 mg were given as the 21-succinate sodium salt in a balanced crossover study to six normal men. Plasma steroid concentrations and blood histamine were measured simultaneously. Both forms of methylprednisolone exhibited linear kinetic parameters. One dynamic model quantitates the baseline circadian pattern and the decline and return of cortisol with similar parameter estimates for all three dose levels. A similar model describes the monoexponential decline and the log-linear return to steady-state baseline of blood histamine. Similar inhibitory concentration values for both effects approximated the equilibrium dissociation constant of in vitro steroid receptor binding. The new models are more physiologically appropriate for these steroid effects than three other models that are commonly employed in pharmacodynamics. Steroid effects generally appear to be receptor mediated with either nongene immediate responses or gene-mediated delayed effects. These models allow quantitation of the rapid effects of steroids with simple equations and common fitted parameters for all steroid dose levels.

Pilot study of the pharmacokinetics of methylprednisolone after single and multiple intravenous doses of methylprednisolone sodium succinate and methylprednisolone suleptanate to healthy volunteers.

The pharmacokinetics of methylprednisolone were evaluated in 29 healthy volunteers after multiple intravenous doses of methylprednisolone sodium succinate or the novel prodrug, methylprednisolone suleptanate. Subjects were assigned randomly to one of four treatment groups (40, 100, 250, or 500 mg) and then randomly assigned to receive either the sodium succinate or suleptanate prodrugs. Doses were administered every 6 hours for 48 hours. Plasma and urine were assayed for methylprednisolone and unchanged prodrug using HPLC methods. Methylprednisolone pharmacokinetics exhibited both a dose and time dependency, which was similar for administration of both prodrugs. After first-dose administration, mean clearance increased from 19.5 L/hr for 40-mg doses to 27.7 L/hr after 500-mg doses of the sodium succinate ester, and from 20.1 to 31.7 L/hr after the suleptanate ester. After multiple dosing, mean clearance values increased from 31.1 to 44.7 L/hr for sodium succinate dosing, and from 31.5 to 46.0 L/hr for suleptanate dosing. Apparent systemic clearance values determined after multiple dosing were 1.5- to 1.8-fold greater than corresponding first-dose values. No dependence on time was apparent for any prodrug pharmacokinetic parameter. These data suggest that the dose dependency of methylprednisolone pharmacokinetics is related to dose-dependent prodrug hydrolysis, whereas the time dependence possibly reflects auto-induction of methylprednisolone metabolism. Based on comparison of methylprednisolone pharmacokinetic parameters derived for each prodrug, methylprednisolone suleptanate resulted in a faster and slightly more efficient conversion to methylprednisolone than methylprednisolone sodium succinate.

Pharmacokinetics of methylprednisolone hemisuccinate and methylprednisolone in chronic liver disease.

The disposition of methylprednisolone (MP) and its prodrug hemisuccinate (MPHS) was assessed in six middle-aged patients with chronic liver disease (CLD) and compared with six younger, healthy subjects after a single IV dose of 25.4 mg of MPHS. Blood and urine samples were collected over 12 hours. Plasma and urine concentrations of MPHS and MP and plasma cortisol were measured by HPLC. MPHS clearance (CL) was significantly reduced in the CLD group (495 vs. 1389 mL/hr/kg) whereas volume of distribution (Vss) of MPHS (about 0.35 1/kg) did not differ. The elimination half-life, t1/2 beta, was significantly longer in CLD (0.61 vs. 0.32 hr). The percent recovery of unchanged MPHS in urine was similar (about 9%) in both groups. The kinetic parameters of MP did not differ between the two groups for: clearance (about 370 L/hr/kg IBW), Vss (about 1.3 L/kg), and t1/2 beta (about 3.0 hr). The suppression t1/2 of cortisol after MPHS was longer (3.9 vs. 1.9 hr) indicating metabolic pathways for cortisol and MP are affected differently in CLD. Reduction in MPHS CL may reflect altered hepatic blood flow due to both cirrhosis and age effects. However, good availability of MP from MPHS and lack of perturbation of MP pharmacokinetics in CLD patients may provide therapeutic advantages in selection of this glucocorticoid. This is the first study that characterizes the disposition of the prodrug MPHS and the formation of MP simultaneously in CLD patients.

Kinetics of hydrolysis of dextran-methylprednisolone succinate, a macromolecular prodrug of methylprednisolone, in rat blood and liver lysosomes.

A macromolecular prodrug of methylprednisolone (MP) was synthesized by conjugating MP with dextran with a M(W) of 70000 through a succinic acid linker. It has been shown previously that the dextran-MP conjugate (DMP) releases MP directly or indirectly through formation of methylprednisolone succinate (MPS) which is further hydrolyzed to MP. To investigate the suitability of DMP conjugate as a prodrug of MP for systemic administration, the kinetics of hydrolysis of the conjugate was studied in vitro in rat blood and liver lysosomes. In blood, the hydrolysis of MPS to MP was approximately ten-fold faster than that in buffer. However, the hydrolysis rate constants of DMP conjugate to MP or MPS in blood were not different from those in buffer. Overall, the hydrolysis of DMP in the rat blood occurred with a half life of approximately 25 h. Hydrolysis of MPS to MP also occurred in the liver lysosomal fraction, but not in the control samples lacking lysosomes. However, the rate constants for the hydrolysis of DMP conjugate to MP and MPS in the lysosomal fraction were not significantly different from those in the control samples. These data suggest that the slow hydrolysis of DMP conjugate to MP or MPS in both rat blood and liver lysosomes occurs mostly, if not completely, via chemical hydrolysis. However, the conversion of MPS to MP is apparently enzymatic. The data may have significant implications for systemic administration of the prodrug.

Pharmacokinetics of methylprednisolone sodium succinate and methylprednisolone in patients undergoing cardiopulmonary bypass.

The pharmacokinetics of methylprednisolone sodium succinate (MPHS) and methylprednisolone (MP) were determined in six patients undergoing open heart surgery with cardiopulmonary bypass. Plasma concentrations of both compounds were measured by high-performance liquid chromatography after doses of MPHS of 1.7-2.4 g. The prodrug ester MPHS yields MP with an average formation rate constant of 0.70 +/- 0.29 hr-1. Peak concentrations of MP occur around 1-2 hours after loading and additional administration of MPHS. The pharmacokinetic values of the two drugs in patients having cardiopulmonary bypass were compared to those in younger, healthy subjects. The volume of distribution of MPHS was lower in the patients, and that of MP was similar to the value in controls. Total clearances of both agents were reduced by about 5 and 2 times. The elimination half-life of MPHS was increased slightly, whereas that of MP increased more than twice in the patients. Significant alterations in clearances occurred in patients, but concentrations of MP were appreciable and prolonged MP due to the extensive formation of MP from MPHS and reduced clearance of MP.

Methylprednisolone sodium succinate in pediatric parenteral nutrition: influence of vehicle injection.

Many drugs can be administered in parenteral nutrient mixtures, but no work on the delivery of corticoids by such means is reported. We studied the influence of pediatric parenteral nutrient mixtures on the kinetic parameters of the corticoid methylprednisolone injected in an intravenous bolus in rabbits as its sodium succinate ester. Four groups of six male New Zealand rabbits were used. After extraction, the plasma drug concentrations were measured by high performance liquid chromatography. The distribution volume of the ester and the clearance rates of the two entities (ester and methylprednisolone) were lowered in the presence of a lipid emulsion. The description of these pharmacokinetic parameters provides a basis for a preclinical study of 24h administration of methylprednisolone in a parenteral nutrient mixture.

Pharmacokinetics of methylprednisolone succinate, methylprednisolone, and lidocaine in the normal dog and during hemorrhagic shock.

Pharmacokinetics of methylprednisolone succinate and methylprednisolone following methylprednisolone sodium succinate administration were studied in five dogs under normal conditions and then during a severe hemorrhagic shock. In order to evaluate hepatic blood flow, lidocaine clearance was simultaneously measured. In the normal state, the clearance of methylprednisolone succinate was 1.64 +/- 0.499 L/h/kg and its half-life was 15.33 +/- 3.84 min. The systemic availability of methylprednisolone from methylprednisolone succinate was 59.9 +/- 8.3%, and the maximal methylprednisolone concentration was observed after a delay of 7.68 +/- 6.31 min. Using a reservoir technique in anesthetized dogs, severe hemorrhagic shock was obtained. Changes in lidocaine clearance indicated a subsequent reduction of hepatic blood flow. The clearance of methylprednisolone succinate decreased to 0.488 +/- 0.240 L/kg/h, and the half-life increased to 40.66 +/- 23.48 min. The exact availability of methylprednisolone from methylprednisolone succinate during shock was not calculable because methylprednisolone kinetics were time dependent. The plasma methylprednisolone concentration was relatively high and persistent during the shock. It was concluded that methylprednisolone sodium succinate is a prodrug which can be released in sufficient quantities as its active moiety (i.e., methylprednisolone) during severe hemorrhagic shock in the dog. In addition, after a single intravenous administration, the slow process of methylprednisolone elimination may give sustained methylprednisolone concentrations for several hours.

Disposition of methylprednisolone and its sodium succinate prodrug in vivo and in perfused liver of rats: nonlinear and sequential first-pass elimination.

The disposition of methylprednisolone (MP) and its prodrug succinate ester, methylprednisolone sodium succinate (MS), were examined both in vivo and in situ (perfused livers) in rats. In vivo studies included iv and oral dosing of 10 or 50 mg/kg of MP in both forms, while liver perfusion involved initial perfusate concentrations of 5 and 25 micrograms/mL of either compound. Steroid concentrations were measured by HPLC. In the intact rat, clearance (CL) values of both compounds were high, twice the hepatic plasma flow, and decreased by one-half after the high dose, indicating nonlinear kinetics. The volumes of distribution of MS and MP were essentially constant with dose. Incomplete availability of MP from iv MS (52-55%) and from the oral dose (10%) was found. Sequential first-pass metabolism was investigated in situ. Extensive hepatic extraction of MP (84%) occurred at the low dose, but decreased to 48% at the high dose, supporting in vivo observations of high CL and nonlinearity. Extraction of MS was also high (83%), but MP availability was slight (8%). The MS and MP data were fitted to a sequential first-pass model yielding an average fraction of MS metabolized-to-MP value of 0.22. The prodrug MS and the active metabolite MP thus demonstrate both systemic and hepatic nonlinearity in rats, and the low availability of MP from iv MS was due, in part, to sequential first-pass elimination. This factor is more extensive in rats than in other species.

Two chromatographic methods for the determination of corticosteroids in human biological fluids: pharmacokinetic applications.

Two techniques, high-performance liquid chromatography (HPLC) and quantitative high-performance thin-layer chromatography coupled with densitometry (HPTLC), were developed for the determination of prednisolone (PL), methylprednisolone (MP), and methylprednisolone sodium succinate (MPSS) in human plasma, saliva, and urine. The HPLC and HPTLC methods shared a single and simple step of an organic extraction procedure and separation of steroids using a normal-phase column or HPTLC plate. The methods allow simultaneous measurement of endogenous cortisol in plasma following the administration of PL and MP. The calibration curves of steroids in all biological fluids were linear over a wide range of concentrations of PL and MP in all biological fluids (0.025-4 micrograms/mL). The limit of detection of both assays for PL and MP was 10 ng/mL in plasma and saliva and 25 ng/mL in urine, and of MPSS was 50 ng/mL in plasma. Both methods were reproducible with an inter- and intra-assay coefficient of variation of less than 10% for all steroids over a wide range of concentrations in all biological fluids. No interference from endogenous steroids was found. The presented methods are simple, rapid, specific, sensitive, reproducible, and economical for the pharmacokinetic study of these steroids. The application of these methods for the pharmacokinetic study of both MP and PL in vivo and the in vitro hydrolysis of MPSS is discussed.

Dextran-methylprednisolone succinate as a prodrug of methylprednisolone: plasma and tissue disposition.

Plasma and tissue disposition of a macromolecular prodrug of methylprednisolone (MP), dextran (70 kDa)-methylprednisolone succinate (DMP), was studied in rats. Single 5-mg/kg doses of DMP or unconjugated MP were administered into the tail veins of different groups of rats (n = 4/group/time point). Blood (cardiac puncture) and tissues (liver, spleen, kidney, heart, lung, thymus, and brain) were collected at various times after DMP (0-96 h) or MP (0-2 h) injections. Concentrations of DMP and MP in samples were analyzed by size-exclusion chromatography (SEC) and reversed-phase high-performance liquid chromatography (HPLC), respectively. Conjugation of MP with 70-kDa dextran resulted in 22-, 300-, and 30-fold decreases in the steady-state volume of distribution, clearance, and terminal plasma rate constant of the steroid, respectively. As for tissue distribution, the conjugate delivered the steroid primarily to the spleen and liver as indicated by 19- and 3-fold increases, respectively, in the tissue/plasma area under the curve (AUC) ratios of the steroid. On the other hand, the tissue/plasma AUC ratios of the prodrug in other organs were negligible. Active MP was released from DMP slowly in the spleen and liver, and AUCs of the regenerated MP in these tissues were 55- and 4.8-fold, respectively, higher than those after the administration of the parent drug. In contrast, no parent drug was detected in the plasma of DMP-injected rats. These results indicate that DMP may be useful for the targeted delivery of MP to the spleen and liver where the active drug is slowly released.

Population pharmacokinetics of methylprednisolone in accident victims with spinal cord injury.

OBJECTIVE: High-dose methylprednisolone (MP) is used to treat acute spinal cord injury (ASCI). The objective of the present study was to determine the pharmacokinetics of the pro-drug methylprednisolone hemisuccinate (MPHS) and MP in accident victims with ASCI. METHODS: The patients (n = 26) were treated with a bolus intravenous loading dose of 30 mg/kg MPHS within 2 h after injury and this was followed by a maintenance infusion of 5.4 mg/kg/h up to 24 h. Blood, CSF and saliva samples were collected up to 48 h after the initial dose and the samples were analyzed by HPLC. Concentration-time data of MPHS and MP were analyzed using population pharmacokinetic analysis with NONMEM software. RESULTS: MPHS and MP could be monitored in plasma and CSF. MP but not MPHS was present in saliva. High variability was seen in the MPHS levels in CSF. The pharmacokinetics of the pro-drug and the metabolite were adequately described by a 2-compartment model with exponential distribution models assigned to the interindividual and the residual variability. At steady state, the average measured MP concentration in plasma was 12.3+/-7.0 microg/ml and 1.74+/-0.85 microg/ml in CSF. The CSF levels of MP could be modeled as a part of the peripheral compartment. CONCLUSION: This study demonstrated that CSF concentrations of MP were sufficiently high after i.v. administration and reflected the concentrations of unbound drug in plasma. Salivary levels of MP were about 32% of the plasma level and may serve as an easily accessible body fluid for drug level monitoring.

In vitro trans-scleral iontophoresis of methylprednisolone hemisuccinate with short application time and high drug concentration.

Trans-scleral iontophoresis, i.e. the application of small electric current to enhance drug transport across sclera is an option for non-invasive delivery of corticosteroids to the posterior segment of the eye. In this paper, in vitro trans-scleral iontophoresis of methylprednisolone hemisuccinate was investigated using concentrated drug solutions and short application times to mimic the iontophoretic conditions of in vivo studies. The drug at the donor concentration of 45 mg/ml was delivered through isolated porcine sclera under passive and iontophoretic conditions (cathodal, 2.4 mA) for 2-15 min. In a second set of experiments, the drug was delivered for 5 min at current intensities of 0.9-7.2 mA. After donor removal, drug release was followed up to 24 h. The exposure of concentrated solutions to sclera for 2-15 min under passive conditions caused a notable accumulation of drug up to 0.8 mg/cm², the release of which was successively followed for 24 h. In cathodal iontophoresis, the amount of accumulated drug increased proportionally to the charge between 0.3 and 1.44 Coulomb. When the charge was increased to 2.16 Coulomb by increasing the application time or current intensity, no further enhancement was recorded. This behaviour can be ascribed to substantial drug adsorption on the scleral tissue, as demonstrated through streaming potential studies, with the consequent increase of the electroosmotic flow that opposes drug transport. The set up suggested here could help in defining the optimal conditions for in vivo studies with animal models and reducing the number of in vivo experiments.

Drug adsorption on bovine and porcine sclera studied with streaming potential.

The affinity of a drug to a biological membrane can affect the distribution and the availability of the active compound to its target. Adsorption is usually determined with in vitro distribution studies based on partitioning of the drug between buffer and tissue, which have limitations such as the high variability of the uptake data and the need for high accuracy in the measurement of drug concentration. Furthermore, distribution studies yield solute concentrations in the bulk of the tissue, whereas electrokinetic phenomena such as streaming potential and electroosmosis reflect the electric charge density on a membrane surface. Streaming potential thus can be used in studying the conditions, by which the charge sign and density can be regulated. That, in turn, has significance to electroosmotic transport mechanism during iontophoresis. In this communication, the adsorption of model compounds methylprednisolone sodium succinate, propranolol, and cytochrome C on bovine and porcine sclera is determined as a function of their concentration by measuring streaming potential. Both membranes had negative streaming potential, proving that they carry negative charge, but had different values at negative and positive pressure differences, which is addressed to the structural asymmetry of these membranes. Bovine sclera had a clearly higher value of streaming potential, ca. -26 nV/Pa, than porcine sclera, ca. -7 nV/Pa (10 mM NaCl solution). All the model compounds were adsorbed on bovine and porcine sclera already in the millimolar concentration range and can have an impact to electroosmosis during transscleral iontophoresis. The results obtained help to better elucidate the phenomena involved in transscleral transport, both in passive diffusion and in iontophoresis, supporting the future application of iontophoresis to the noninvasive delivery of drugs to the posterior segment of the human eye.

Fabrication principles and their contribution to the superior in vivo therapeutic efficacy of nano-liposomes remote loaded with glucocorticoids.

We report here the design, development and performance of a novel formulation of liposome- encapsulated glucocorticoids (GCs). A highly efficient (>90%) and stable GC encapsulation was obtained based on a transmembrane calcium acetate gradient driving the active accumulation of an amphipathic weak acid GC pro-drug into the intraliposome aqueous compartment, where it forms a GC-calcium precipitate. We demonstrate fabrication principles that derive from the physicochemical properties of the GC and the liposomal lipids, which play a crucial role in GC release rate and kinetics. These principles allow fabrication of formulations that exhibit either a fast, second-order (t(1/2) ~1 h), or a slow, zero-order release rate (t(1/2) ~ 50 h) kinetics. A high therapeutic efficacy was found in murine models of experimental autoimmune encephalomyelitis (EAE) and hematological malignancies.