Enhancement of adriamycin antitumor activity by its binding with an intracellular sustained-release form, polymethacrylate nanospheres, in U-937 cells.

We investigated the antitumor activity of Adriamycin on a monocytic-like cancer cell line U-937 after its binding on polymethacrylate nanospheres (diameter, 270-350 nm). Compared to free Adramycin (F-ADR), nanosphere-bound Adriamycin (B-ADR) exhibits a 3-fold enhancement of cytotoxicity, as determined by cell growth inhibition and DNA synthesis, after continuous exposure to 0.02 and 0.04 microgram/ml. The 90% growth inhibition concentration was 0.051 microgram/ml for F-ADR and was 0.018 microgram/ml for B-ADR (P less than 0.001). Furthermore, the nanosphere densities per cell play an important role since for the same drug concentration the higher the density increases, the better the activity is. Indeed, after 4 days of incubation in a medium containing 160 nanospheres at 0.5 fg/cell, the cell counts were 62.8 +/- 12.8% (SD) of the initial inoculum and they were only 16.1 +/- 0.1% after incubation in a medium containing 800 nanospheres at 0.1 fg/cell (P less than 0.001). A comparable enhancement of activity regarding the nanosphere densities was observed after a 24-h exposure to 0.02 and 0.05 microgram/ml. Short-term uptake studies showed that B-ADR accumulation was higher with B-ADR than with F-ADR. In addition, the efflux kinetics was modified. For cells exposed to F-ADR for 4 h, the efflux half-life was 23.7 +/- 7.7 h and the area to infinity under the efflux curve was 8.6 +/- 2.8 micrograms/mg protein x h-1. For cells exposed to B-ADR, the efflux half-life increased to 85.9 +/- 19.2 h and the area to infinity under the efflux curve to 29.6 +/- 6.6 micrograms/mg protein x h-1 (P less than 0.001). Electron transmission microscopy and previous findings have revealed that B-ADR was well internalized into cells. Our data support the hypothesis that B-ADR acts as an intracellular drug release complex after endocytosis. The findings regarding the number of nanospheres per cell and dose-effect relationships are consistent with mechanisms of drug actions extending to membrane domains.

Ophthalmic drug delivery systems--recent advances.

Eye-drops are the conventional dosage forms that account for 90% of currently accessible ophthalmic formulations. Despite the excellent acceptance by patients, one of the major problems encountered is rapid precorneal drug loss. To improve ocular drug bioavailability, there is a significant effort directed towards new drug delivery systems for ophthalmic administration. This chapter will focus on three representative areas of ophthalmic drug delivery systems: polymeric gels, colloidal systems, cyclodextrins and collagen shields. Hydrogels generally offer a moderate improvement of ocular drug bioavailability with the disadvantage of blurring of vision. In situ activated gel-forming systems are preferred as they can be delivered in drop form with sustained release properties. Colloidal systems including liposomes and nanoparticles have the convenience of a drop, which is able to maintain drug activity at its site of action and is suitable for poorly water-soluble drugs. Among the new therapeutic approaches in ophthalmology, cyclodextrins represent an alternative approach to increase the solubility of the drug in solution and to increase corneal permeability. Finally, collagen shields have been developed as a new continuous-delivery system for drugs that provide high and sustained levels of drugs to the cornea, despite a problem of tolerance. It seems that new tendency of research in ophthalmic drug delivery systems is directed towards a combination of several drug delivery technologies. There is a tendency to develop systems which not only prolong the contact time of the vehicle at the ocular surface, but which at the same time slow down the elimination of the drug. Combination of drug delivery systems could open a new directive for improving results and the therapeutic response of non-efficacious systems.

Impact of long-term ethanol consumption on CYP1A2 activity.

Ethanol is a well-known inducer of CYP2E1; whether or not it is an inducer of other cytochromes has not been investigated systematically. The aim of our study was to evaluate the impact of ethanol consumption on the activity of CYP1A2, which has been shown to be influenced by drugs (inhibited or induced). We evaluated CYP1A2 activity by the ratio of the molar urinary concentrations of the three end products of paraxanthine demethylation of caffeine to the molar concentration of a paraxanthine 8-hydroxylation product. This urinary metabolite ratio has previously been shown to correlate with caffeine clearance. The caffeine metabolites were measured in urine collected during the 3 hours after oral administration of 200 mg caffeine. The caffeine test was performed in 12 smokers (> 25 cigarettes/day) and 12 nonsmokers, all of whom were alcoholic inpatients (daily intake > 100 mg absolute ethanol), within the first 3 days of their hospital stay and after 14 days of abstinence from ethanol. In alcoholic patients who were smokers the molar urinary concentration ratio was 3.14 +/- 0.97 before withdrawal and 4.01 +/- 0.92 after 14 days of abstinence from ethanol. In contrast, in alcoholic patients who were nonsmokers it was 2.62 +/- 0.95 and 2.18 +/- 0.96 before and after withdrawal, respectively. In volunteers who were smokers the molar urinary concentration ratio was 5.02 +/- 1.51, whereas in volunteers who were nonsmokers it was 3.22 +/- 1.46. Our results confirm the well-known induction of CYP1A2 activity by tobacco smoking and show that this induction is masked by long-term ethanol consumption.

A dose-response study of epidural liposomal bupivacaine in rabbits.

Liposomes are drug delivery systems used to prolong local effects of bupivacaine. We studied the relationships between motor and hemodynamic changes and epidural doses of plain bupivacaine (P) and liposomal bupivacaine (L) in rabbits equipped with chronical lumbar epidural and femoral arterial catheters. Liposomal (phosphatidylcholine-cholesterol) suspensions contained 20 mg ml-1 of lipid, and different doses of bupivacaine (Lipo 7.5=7.5-; Lipo 3.7=3. 75-; Lipo 2.5=2.5-; Lipo 1.2=1.25-; and Lipo 0.7=0.65-mg of bupivacaine per ml). Forty rabbits were randomly assigned to five groups to receive epidural anesthesia (1 ml) as follows: Groups I to V received 0.65 to 7.5 mg of bupivacaine as P then as L. Release rate of bupivacaine from liposome was significantly slower using Lipo 3.7 than after Lipo 2.5 (Td was 3.9 h and 1.7 h respectively). Increasing the doses of L and P resulted in faster onset time for complete motor blockade and in a prolonged duration of motor effects. Liposomal formulation appears to be a powerful delivery system to prolong the motor effects of bupivacaine since E50 was lower and Emax higher than after the use of plain solution (E50 4.49+/-1.81 mg and Emax 152+/-40 min for P; and E50 2.61+/-0.23 mg and Emax 202+/-9 min for L). Hemodynamic changes were linearly related to doses of bupivacaine injected. The best bupivacaine-to-lipid ratio to prolong motor effects using our model was 3.75 mg and 20.0 mg respectively (Lipo 3.7).

A comparison of radiopharmaceutical agents used for the diagnosis of pulmonary embolism.

Radioactive gas or technetium-99m aerosols are used to perform pulmonary ventilation scintigraphy. The aim of this study was to compare three radiopharmaceuticals, Kryptoscan, Technegas and Venticis II, in terms of their costs and user preferences rather than on the basis of diagnostic efficacy. For each radiopharmaceutical agent, an analysis questionnaire was sent to nuclear medicine departments setting out the criteria (and subcriteria) to be assessed: diagnosis quality: imaging quality, distribution homogeneity, examination procedures and capacity to examine particular patients (e.g. smokers); safety: for patient, paramedical and medical staff and the environment; use: availability in cases of emergency, ergonomics of the apparatus, simplicity and time of preparation. A score, ranging from 0 to 5, and a weighting (importance of one criterion with regard to the others) were assigned to each criterion. The direct cost of a ventilation (drugs, generator systems, disposable materials) was calculated for each radiopharmaceutical agent according to the number of patients examined per day (1-6) and the number of examination days per week (2-5). Fourteen questionnaires concerning at least two of the products were returned out of the 30 mailed. A 'preference score' was calculated using Pharma Decision software. The mean score of Kryptoscan was significantly higher than that of Venticis II (444 vs. 286, P < 0.001) and higher than the mean score of Technegas (444 vs. 344, P < 0.01). For Venticis II and Technegas, the changes in patient direct costs were minor and depended on the number of patients per day and the number of examination days per week. Respectively, they were: $US 117.66 (5 patients.day-1; 5 days.week-1) to $US 147.74 (2 patients.day-1; 2 days.week-1) and $US 56.60 (6 patients.day-1; 5 days.week-1) to $US 132.08 (2 patients.day-1; 2 days.week-1). The direct cost of ventilation using Kryptoscan varied only according to the number of patients examined per day: $US 104.66 (6 patients.day-1) to $US 266.47 (2 patients.day-1). This study shows that Kryptoscan appears to be preferable for ventilation scintigraphy whenever at least four patients are examined daily.

Preparation and in vivo studies of a new drug delivery system. Nanoparticles of alkylcyanoacrylate.

Polyhexylcyanoacrylate nanoparticles have been prepared with vincamine as the model drug. These particles had an average size of 200 nm and adsorbed approximately 43% of vincamine. The adsorption of vincamine to nanoparticles modified the distribution of vincamine in tissues. After iv injection the distribution volumes were increased in comparison with an aqueous solution of drug. In comparison with an aqueous solution of drug, the absolute bioavailability of vincamine was also increased after an oral administration of nanoparticles.

[Biopharmaceutics of local anesthetic-cyclodextrin complexes following loco-regional administration]. / Biopharmacie de complexes anesthésiques locaux-cyclodextrines après administration loco-régionale.

In the research of new dosage forms improving the therapeutic index of local anesthetics, we studied cyclodextrins, cyclic oligosaccharides forming soluble inclusion complexes with various lipophilic drugs. Complexes between bupivacaine and hydroxypropyl-B-cyclodextrin, bupivacaine and sulfobutyl ether-7-B-cyclodextrin were studied in vivo, using an epidural and a perineural (sciatic) model, respectively. Biopharmaceutics and pharmacodynamics of bupivacaine were evaluated in the rabbit. In both models, only systemic absorption rate of bupivacaine was decreased upon complexation, not the quantity absorbed. Complexation with cyclodextrins could be a promising drug delivery system to improve the therapeutic index of local anesthetics.

[The effect of modification of excipients for the bioavailability of vinburnine]. / Influence d'une modification d'excipients sur la biodisponibilité de la vinburnine.

Biopharmaceutic comparison is achieved between commercialized capsules of vinburnine and a new dosage form. The difference between the two drug products concerns only the quality of excipients. Drug bioavailability decreases of about 10%, in despite of the very important, and well known, interindividual variations of vincamine drug products. Only the absorption rate seems statistically lightly decreased by the new formulation.

[The oral liquid form of vinburnine: biopharmaceutics]. / Forme buvable de vinburnine: une approche biopharmaceutique.

Biopharmaceutic comparison is achieved between commercialized capsules of vinburnine and drops dosage form intended for geriatric pharmacotherapy. Drug bioavailability of vinburnine seems saved, if not significantly increased by the new formulation, in spite of the very important, and well known interindividual variations of vincamine drug products. Only the absorption rate seems statistically increased by drops.

[Simultaneous determination of vinburnine and 6-hydroxyvinburnine in human plasma by high-performance liquid chromatography]. / Dosage simultané de la vinburnine et de la 6-hydroxyvinburnine par chromatographie liquide à haute performance.

An isocratic high-performance liquid chromatographic method has been developed to allow the simple and rapid determination of both vinburnine (I) and its main metabolite, 6-hydroxyvinburnine (II), in heparinized human plasma (0.5 ml). Compounds I and II and p-chlorodisopyramide (internal standard) were first extracted with alkalinized ethyl acetate and then with sulphuric acid. Separation was achieved on a reversed-phase muBondapak C18 column with a mobile phase of acetonitrile-water-0.1 M heptanesulphonate in acetic acid and with detection at 254 nm. Each run required 20 min. The within-day coefficients of variation for identical samples (20 ng/ml) were 7 and 6% and between-day coefficients of variation 8 and 26% for I and II, respectively. The detection limit was 5 ng/ml (normal therapeutic concentration, 10-300 ng/ml). The application of the method to drug monitoring was compared to that of a thin-layer chromatographic procedure.

Prolongation of spinal anesthesia with bupivacaine-loaded (DL-lactide) microspheres.

There is considerable interest in developing a sustained-release local anesthetic formulation to provide long-lasting anesthesia and to decrease systemic toxicity. Bupivacaine (B), 10 mg, loaded in two different types of polylactide microspheres (PLA1 and PLA2) was evaluated after spinal injection and compared with plain bupivacaine (pB), 2 mg. Experiments were performed in six New Zealand rabbits. Duration of motor block was significantly prolonged for PLA1 compared to pB (177.5 +/- 79.5 min vs 44.6 +/- 18.0 min; P < 0.05), as well as for the recovery time (545.0 +/- 299.6 min vs 44.2 +/- 21.5 min; P < 0.05). The duration and recovery were not prolonged for PLA2. Systemic release of B after intrathecal injection was measured from blood samples by using high-performance liquid chromatography. There was no significant difference in maximum B plasma concentration between pB and PLA1 (326 +/- 81 mg/mL vs 321 +/- 57 ng/mL). The time taken to reach the maximum plasma concentration (6.6 +/- 2.6 min vs 41.7 +/- 20.4 min; P < 0.05) was significantly different. This study demonstrated that the use of bupivacaine-loaded (DL-lactide) microspheres can prolong spinal motor block.

Bioavailability of oral vs intramuscular iodinated oil (Lipiodol UF) in healthy subjects.

BACKGROUND: In order to fight against iodine deficiency, the essential cause of endemic goiter and cretinism, several health organizations promoted campaigns of iodinated oil (Lipiodol UF) administration using iodinated oil administered intramuscularly. However, it seems preferable to administer iodinated oil orally, as this is more appropriate and since the efficacy of this route has been demonstrated as well as for intramuscular route by controlled clinical trials. OBJECTIVE: To assess the bioavailability of iodinated oil (Lipiodol UF) administered via two different administration routes and the safety profile of this agent. DESIGN: A randomized bioavailability study was performed comparing a single oral dose of 3 capsules (570 mg of iodine) vs a single intramuscular injection of 1 ml of Lipiodol UF (480 mg of iodine) in 36 healthy subjects followed for 9 months. RESULTS: The results show that, at these dosages, the 24 h urinary iodine values are above baseline for both oral and intramuscular administrations (im: >12 months/oral: 6 months) for prolonged period of time. In terms of safety, Lipiodol, administered by im injection or orally, did not induce any undesirable effects or any alteration of thyroid function tests in this study. CONCLUSIONS: In conclusion, this study shows that im or oral administration of a single dose of Lipiodol provides a significant and prolonged iodine supplement. The results obtained confirm the possibility of protection of exposed populations after annual administration of an appropriate single oral dose, without inducing any clinical or laboratory adverse effects. The product, by either route of administration, has a prolonged efficacy in iodine-deficient subjects (im: 2-3 years/oral: 1 year).

Effects of different formulations of mitoxantrone (solutions, nanospheres, liposomes) on glaucoma surgery in rabbits.

Pharmacological blockade of fibroblastic proliferation after glaucoma filtration surgery by using antimitotic agents in different formulations (liposomes, nanospheres) is of great clinical interest. However, only limited comparative data are available on the effect of encapsulated drugs on intraocular pressure (IOP) after filtering surgery. Therefore we have studied the effect on IOP of liposomes, nanospheres and a solution of mitoxantrone (MTO), a well-known antimitotic, in rabbits before and after sclerectomy. MTO in solution form, as well as in a liposome formulation, improved the outcome of the surgery by reducing IOP when administered subconjunctivally just following surgery. This effect was similar to mitomycin-C application. In contrast, neither prior administration nor subconjunctival nanosphere injections induced a reduction in IOP. Liposome administration demonstrated no delayed action or promoting effect but reduced the occurrence of corneal opacity observed in groups treated with MTO in solution.

Effect of cyclosporine A formulations on bovine corneal absorption: ex-vivo study.

The purpose of this study was to evaluate, in an ex-vivo study, the absorption of cyclosporine A on bovine cornea after 24 h contact with various drug delivery systems containing 1% cyclosporine A and in comparison with an olive oil formulation as the reference vehicle for cyclosporine A. The different formulations studied were poly(acrylic acid) polymeric gels in aqueous/non-aqueous solvents, polyisobutylcyanoacrylate nanocapsules, and a combination of both formulations. The histological effects of these formulation on corneal cells after 24 h of contact were also studied. The lowest absorption rate of cyclosporine A was found using olive oil with a percent absorption of 2.52 +/- 1.52% (259 +/- 171 micrograms/g cornea). The three formulations developed for this study, nanocapsules, poly(acrylic acid) polymeric gel and nanocapsules gel showed significantly better absorption of CsA than olive oil, with a mean percent absorption of 5.81 +/- 2.04% (621 +/- 218 micrograms/g cornea), 6.09 +/- 2.93% (651 +/- 313 micrograms cornea) and 7.92 +/- 2.55% (847 +/- 273 micrograms/g cornea) respectively. As we studied the penetration of cyclosporine A into the different layers of the cornea, we observed that for all formulations, CsA remained at the corneal surface and did not penetrate the whole cornea. The histological study showed that olive oil, nanocapsules and poly(acrylic acid) gel in aqueous/non-aqueous solvents show some modifications on the cornea, contrary to the nonocapsules gel which did not indicate any toxic effect. The nanocapsule gel, with the highest percent absorption along with its margin of safety on the cornea, seems to present a new promising drug delivery system for ocular administration.

Release kinetics of liposome-encapsulated ganciclovir after intravitreal injection in rabbits.

This study was undertaken to establish experimentally whether the intravitreal application of liposomally-entrapped ganciclovir could prolong intraocular therapeutic levels when it is compared to the intravitreal injection of a simple solution of the drug. New Zealand white rabbits were given an intravitreal injection of the drug solution and of liposome-encapsulated ganciclovir. The intravitreal clearance of ganciclovir was determined after a single injection of either the drug solution (200 micrograms/0.1 mL) or the liposomally-entrapped (with 41% load; 82 micrograms drug load and 118 micrograms free) ganciclovir. The ganciclovir vitreal concentrations were measured at various time intervals for a period up to 43 days using an HPLC method. The results of this study clearly demonstrated that prolonged intravitreal drug levels (above the mean inhibitory dose of cytomegalovirus of 1 microgram/mL) after administration of the liposome-entrapped ganciclovir and estimated to continue beyond 30-43 days. The injection of the 200 micrograms/0.1 mL of drug solution showed a mean vitreous concentration which was higher than the ID50 only for 55 h. The disappearance rate constant for the liposome-encapsulated injections was approximately 22 x slower than simple drug solution injections (controls). No evidence of retinal toxicity was found by clinical or light microscopy examination of the treated eyes.