Safety,efficacy and toxicological evaluation of a novel,patented anti-diabetic extract of Trigonella Foenum-Graecum seed extract (Fenfuro)

Safety and anti-diabetic efficacy of a novel, proprietary Trigonella foenum-graecum seed extract [novel fenugreek extract (FE), Fenfuro?, CR0010810) enriched in furostanolic saponins (>60% w/w, HPLC) were assessed. Concerning safety, we undertook studies dealing with acute oral toxicity, 28-d sub-chronic toxicity and Ames’ bacterial reverse mutation assay that revealed no toxicity. Concerning efficacy, we examined beneficial effects of the extract on rats with type 2 diabetes (T2D). Male Sprague–Dawley rats received a high-fat diet for 2 weeks followed by streptozotocin (STZ, 35?mg/kg i.p.) to produce T2D. Seven days post-STZ, rats showing ≥300?mg/dl fasting plasma glucose level (PGL) were included in the study. FE (150- or 450- mg/kg p.o.) and glipizide (5?mg/kg p.o.) were administered once daily for 20?d and then twice daily for another 10?d (total 30?d). Blood samples were collected at 0, 10, 20 and 30?d of treatment and estimated for fasting plasma triglyceride (PTG), total cholesterol and insulin levels. After 30?d, FE and glipizide-treated diabetic animals were treated in combination with or without metformin (100?mg/kg) twice daily for another 10?d. FE did not influence body weight, feed and water intake. FE (150?mg/kg p.o.) reduced PTG levels in T2D rats by 22%, 24.6% and 29% at 10, 20 and 30?d of treatment, respectively, while glipizide (5?mg/kg p.o.) reduced the PTG levels by 57.4%, 46.2% and 39.4% at these time points. FE (450?mg/kg) treatment in STZ-induced diabetic rats produced significant hypoglycemic activity (approximately 31.5%) as compared to insulin (48.2% with 1 U/kg i.p.). FE (150?mg/kg p.o.) and metformin (100?mg/kg p.o.) combined produced significant reduction (20.7%) of PGL in T2D rats. No adverse effects were observed. We conclude after extensive in vitro and in vivo safety and efficacy studies that FE is safe and effective in treating T2D.     

Analgesic,anti-inflammatory,antipyretic and toxicological evaluation of some newer 3-methyl pyrazolone derivatives

In this paper, we described the pharmacological and toxicological studies of three pyrazolone derivatives namely PYZ1: 4-[4-N dimethylamino benzylidine]-3-methyl pyrazolin-5(4H)-one, PYZ2: 4-[2-chlorobenzylidine]-3-methylpyrazolin-5(4H)-one and PYZ3: 4-[benzylidine]-3-methylpyrazolin-5(4H)-one derivatives. Analgesic, anti-inflammatory and antipyretic studies of 3-methyl pyrazolone derivatives at 400 mg/kg, p.o. have shown significant activity as compared to control. Amongst three pyrazolone derivatives, PYZ2 was found to be more active. Based on the result of pharmacological studies, PYZ2 was selected for toxicological studies. Acute toxicity studies revealed that methyl pyrazolone derivatives are non-toxic in rats up to 5000 mg/kg, p.o. The subacute toxicity study of PYZ2 showed that decrease in Hb content, RBC and WBC count. In biochemical analysis level of blood glucose and bilirubin reduced where as AST, ALT and alkaline phosphatase level elevated. Histopathological studies revealed that there was mild toxicity on liver and kidney at 1000 mg/kg, p.o.     

Safety and toxicological evaluation of a novel,water-soluble undenatured type II collagen

Abstract

This study was conducted to determine the broad-spectrum safety of a novel, water-soluble undenatured type II collagen (NEXT-II) derived from chicken sternum cartilage. The presence of epitope in NEXT-II was confirmed by using a commercial kit. The acute oral LD₅₀ of NEXT-II was found to be greater than 5000?mg/kg bw in rats, while the single-dose acute dermal LD₅₀ was greater than 2000?mg/kg bw. The primary dermal irritation index (PDII) of NEXT-II was found to be 1.8 and classified as slightly irritating to the skin. In primary eye irritation studies, the maximum mean total score (MMTS) of NEXT-II was observed to be 7.3 and classified as minimally irritating to the eye. Long-term safety studies were conducted in dogs over a period of 150?d, and no significant changes were observed in body weight, heart rate, respiration rate and blood chemistry. NEXT-II does not induce mutagenicity in the bacterial reverse mutation test in five Salmonella typhimurium strains either with or without metabolic activation. Furthermore, two experiments were conducted to assess the potential of NEXT-II to induce mutations with and without metabolic activation at the mouse lymphoma thymidine kinase locus using the cell line L5178Y. No biologically relevant increase of mutants was observed. Also, no dose-dependent toxicity was observed. Furthermore, colony sizing showed no clastogenic effects induced by NEXT-II under the experimental conditions. These studies demonstrated the broad spectrum of safety of NEXT-II.     

Toxicological safety evaluation of a novel highly bioavailable turmeric extract formulation

Turmeric (Curcuma longa L.) extracts have a long history of use worldwide, but a major limitation of these extracts is their extremely low oral bioavailability, caused by low absorption, rapid metabolism and rapid excretion following ingestion. Thus, a new highly bioavailable turmeric extract formulation (comprising turmeric extract, acacia gum, sunflower oil and quillaia extract) has been developed and is intended for use as a food ingredient. Safety of this novel extract was evaluated using the standard Tier 1 battery of in vitro genotoxicity tests (bacterial reverse mutation test and an in vitro mammalian cell micronucleus test) followed by repeated-dose 28- and 90-day oral toxicity studies in rats. In the 90-day study, male and female Sprague-Dawley rats were dosed once daily, by oral gavage, either with the vehicle or the test item at 500, 1500 or 3000 mg/kg body weight/day. Clinical examinations were conducted regularly, and body weights and food consumption were recorded weekly throughout the study. At the end of the study, blood samples were analyzed for clinical pathology parameters, before a macroscopic necropsy was conducted and a full list of tissues were examined histopathologically. There was no evidence of genotoxicity in vitro. No test item-related adverse effects were observed in the 28- or 90-day studies; therefore, 3000 mg/kg body weight/day (the maximum feasible dose and highest dose tested in rats) was established as the no-observed-adverse-effect level.     

Potential antioxidant,anticholinergic, antidiabetic and antiglaucoma activities and molecular docking of spiraeoside as a secondary metabolite of onion (Allium cepa)

Onion contains many dietary and bioactive components including phenolics and flavonoids. Spiraeoside (quercetin-4-O-β-D-glucoside) is one of the most putative flavonoids in onion. Several antioxidant techniques were used in this investigation to assess the antioxidant capabilities of spiraeoside, including 1,1-diphenyl-2-picrylhydrazyl radical (DPPH·) scavenging, N,N-dimethyl-p-phenylenediamine radical (DMPD^•+) scavenging, 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulphonate) radical (ABTS^•+) scavenging activities, cupric ions (Cu²⁺) reducing and potassium ferric cyanide reduction abilities. In contrast, the water-soluble α-tocopherol analogue trolox and the conventional antioxidants butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), and α-tocopherol were utilized as the standards for evaluation. Spiraeoside scavenged the DPPH radicals an IC₅₀ of 28.51 μg/mL (r²: 0.9705) meanwhile BHA, BHT, trolox, and α-tocopherol displayed IC₅₀ of 10.10 μg/mL (r²: 0.9015), 25.95 μg/mL (r²: 0.9221), 7.059 μg/mL (r²: 0.9614) and 11.31 μg/mL (r²: 0.9642), accordingly. The results exhibited that spiraeoside had effects similar to BHT, but less potent than α-tocopherol, trolox and BHA. Also, inhibitory effects of spiraeoside were evaluated toward some metabolic enzymes including acetylcholinesterase (AChE), butyrylcholinesterase (BChE), carbonic anhydrase II (CA II) and α-glycosidase, which are related to a number of illnesses, such as Alzheimer’s disease (AD), diabetes mellitus and glaucoma disorder. Spiraeoside exhibited IC₅₀ values of 4.44 nM (r²: 0.9610), 7.88 nM (r²: 0.9784), 19.42 nM (r²: 0.9673) and 29.17 mM (r²: 0.9209), respectively against these enzymes. Enzyme inhibition abilities were compared to clinical used inhibitors including acetazolamide (for CA II), tacrine (for AChE and BChE) and acarbose (for α-glycosidase). Spiraeoside demonstrated effective antioxidant, anticholinergic, antidiabetic and antiglaucoma activities. With these properties, it has shown that Spiraeoside has the potential to be a medicine for some metabolic diseases.     

Toxic essential oils. Part II: Chemical,toxicological, pharmacological and microbiological profiles of Artemisia annua L. volatiles

Botanical drugs based on Artemisia annua L. (Asteraceae) are important in the treatment of malaria. Alongside with artemisinin, this aromatic species produces high and variable amounts of other chemicals that have mostly unknown biological/pharmacological activities. Herein, we have studied the toxicological/pharmacological profile of volatile constituents of a Serbian population of A. annua. Fifty-eight components were identified, among them, artemisia ketone (35.7%), α-pinene (16.5%) and 1,8-cineole (5.5%) were the most abundant ones. Significant variability of A. annua volatile profile was confirmed by means of agglomerative hierarchical cluster analysis indicating the existence of several different A. annua chemotypes. In an attempt to connect the chemical profile of A. annua oil with its biological/toxicological effects, we have evaluated in vivo and/or in vitro toxicity (including hepato- and nephrotoxicity/protection), antinociceptive, antioxidant (DPPH, ABTS and superoxide radical scavenging activity assays), enzyme inhibiting (protein kinase A and α-amylase) and antimicrobial potential of A. annua oil and of its constituents. Our results revealed that the beneficial properties of A. annua botanical drugs are not limited only to their antimalarial properties. Taking into account its relatively low toxicity, the usage of A. annua volatiles (at least of the herein studied population) does not represent a health risk.     

Safety and toxicological evaluation of a synthetic vitamin K2, menaquinone-7

Menaquinone-7 (MK-7) is part of a family of vitamin K that are essential co-factors for the enzyme γ-glutamyl carboxylase, which is involved in the activation of γ-carboxy glutamate (Gla) proteins in the body. Gla proteins are important for normal blood coagulation and normality of bones and arteries. The objective of this study was to examine the potential toxicity of synthetic MK-7 in BomTac:NMRI mice and in Sprague-Dawley rats. In an acute oral toxicity test, mice were administered a single oral dose of 2000?mg/kg body weight (limit dose) and no toxicity was observed during the 14-day observation period. In the subchronic oral toxicity test in rats, animals were administered MK-7 for 90 days by gavage at the following doses: 0 (vehicle control, corn oil), 2.5, 5, and 10?mg/kg body weight/day. All generated data, including clinical observations, ophthalmology, clinical pathology, gross necropsy, and histopathology, revealed no compound-related toxicity in rats. Any statistically significant findings in clinical pathology parameters and/or organ weights noted were considered to be within normal biological variability. Therefore, under the conditions of this experiment, the median lethal dose (LD₅₀) of MK-7 after a single oral administration in mice was determined to be greater than the limit dose level of 2000?mg/kg body weight. The no observed adverse effect level (NOAEL) of MK-7, when administered orally to rats for 90 days, was considered to be equal to 10?mg/kg body weight/day, the highest dose tested, based on lack of toxicity during the 90-day study period.     

Toxic effects of nitrogenous compounds (ammonia,nitrite, and nitrate) on acute toxicity and antioxidant responses of juvenile olive flounder,Paralichthys olivaceus

Juvenile Paralichthys olivaceus (mean length 7.29 ± 0.59 cm, mean weight 2.41 ± 0.35 g) were exposed to several concentrations of ammonia (0, 6.25, 12.5, 25, 50, and 100 mg/L), nitrite (0, 50, 100, 200, 400, and 800 mg/L), and nitrate (0, 250, 500, 1000, 2000, and 4000 mg/L) for 96 h in 20-L glass tanks. Lethal concentration 50% (LC₅₀) was determined after removing and counting dead fish at 0, 3, 6, 12, 24, 48, 72, and 96 h of exposure. Exposure was significantly toxic to P. olivaceus, and LC₅₀ at 96 h was 26.008 mg/L for ammonia, 768.078 mg/L for nitrite, and 1431.343 mg/L for nitrate. The toxicity profile found for P. olivaceus juveniles was ammonia > nitrite > nitrate. For antioxidant activity analysis such as superoxide dismutase (SOD) and catalase (CAT) activity, liver and kidney tissues were dissected after 96 h of exposure. In liver and kidney tissues, SOD activity was significantly increased at 25 mg/L of ammonia, above 400 mg/L of nitrite, and at 1000 mg/L of nitrate. At these concentrations, CAT activity also increased, except in the kidney, where no change in CAT activity was detected under exposure to nitrate. The results of this study suggest that exposure to nitrogenous compounds such as ammonia, nitrite, and nitrate can induce significant toxicity and alterations in the antioxidant responses of P. olivaceus.     

Evaluation of a new coprocessed compound based on lactose and maize starch for tablet formulation

The development of new direct compression excipients should include a comprehensive and rapid determination of deformation properties. The aim of this study was to characterize StarLac, a new coprocessed compound for direct compression based on lactose and maize starch. For this purpose, the effects of the base materials (maize starch and spraydried lactose) were considered and the influence of the spray-drying process was investigated. This was performed by comparing the physical mixture of starch and spray-dried lactose at the same ratio as for StarLac. For analysis of the deformation behavior, the 3-D model and the Walker equation were applied; for verification, the Heckel equation and the pressure time function (a modified Weibull equation) were used. The advantages of StarLac are its good flowability depending on the spray-drying process, an acceptable crushing force due to its lactose content, its rapid disintegration depending on starch, and a brilliant fast release of an active ingredient, such as theophylline monohydrate. The volume-pressure deformation properties of StarLac were dependent on the lactose properties. Only at high maximum relative density (varrho(rel,max)) did the influence of starch cause a change in these properties. A network-like structure can be observed using scanning electron microscopy pictures. Overall, StarLac deformed plastically with a low portion of elasticity. The physical mixture exhibited a more elastic behavior than StarLac. However, the part of the powder that was irreversibly compressed was much lower than was observed for the single substances. This behavior is caused by an interaction between the components, which in StarLac is prevented by spray drying.     

Influence of the spray adjuvant on the toxicity effects of a glyphosate formulation

In the present study, the influence of the spray adjuvant on the toxicity effects of a glyphosate formulation was examined in HEp-2 cell line. We determined the median lethal concentration (LC₅₀) of Atanor® (glyphosate formulation), Impacto® (spray adjuvant) and the mixture of both agrochemicals. We also compared the toxicities of the pesticides individually and in mixture and we analyzed the effects on oxidative balance from each treatment.Our results showed that all the agrochemicals assayed induce dose and time-dependent cytotoxicity and that the toxicity of Impacto® with Atanor® (mixture) was additive on HEp-2 cell line.All the agrochemicals assayed produced an increase in catalase activity and glutathione levels, while no effects were observed for superoxide dismutase and glutathione-S-transferase activities. We found an important increase in ROS production in cells treated with Atanor® and mixture. Besides, all the agrochemicals used triggered caspase 3/7 activation and hence induced apoptosis pathway in this cell line. In conclusion, our results demonstrated that the addition of adjuvant to glyphosate formulation increase the toxicity of the mixture in cell culture. Furthermore, cell culture exposed to agrochemical mixture showed an increased ROS production and antioxidant defenses.     

Safety assessment of methanol extract of red dragon fruit (Hylocereus polyrhizus): Acute and subchronic toxicity studies

Recently, the fruits of Hylocereus polyrhizus, known as red dragon fruit, have received much attention from growers worldwide. However, there is little toxicological information regarding the safety of repeated exposure to these fruits. The present study evaluated the potential toxicity of a methanol extract of H. polyrhizus fruit after acute and subchronic administration in rats. In the acute toxicity study, single doses of fruit extract (1250, 2500 and 5000 mg/kg) were administered to rats by oral gavage, and the rats were then monitored for 14 days. In the subchronic toxicity study, the fruit extract was administered orally to rats at doses of 1250, 2500 and 5000 mg/kg/day for 28 days. There was no mortality or signs of acute or subchronic toxicity. There was no significant difference in body weight, relative organ weight or hematological parameters in the subchronic toxicity study. Biochemical analysis showed some significant changes, including creatinine, globulin, total protein and urea levels. No abnormality of internal organs was observed between treatment and control groups. The lethal oral dose of the fruit extract is more than 5000 mg/kg and the no-observed-adverse-effect level (NOAEL) of the extract for both male and female rats is considered to be 5000 mg/kg per day for 28 days.     

Hepatoprotective activity of Balsamodendron mukul extract against Paracetamol-induced liver toxicity in rats: In vivo pharmacological and toxicological evaluation

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Gastroscopic and pharmacokinetic evaluation of a new pivmecillinam tablet

Summary Three different pivmecillinam preparations, a conventional 200 mg tablet (P tablet) and two new formulations containing respectively pivmecillinam 200 mg and 400 mg plus Avicel^? (microcrystalline cellulose) as a disintegrator (PA tablet), were compared in vitro and in a gastroscopic study in 8 healthy volunteers. Disintegration of the PA tablet was significantly more rapid both in vitro and in the stomach. Following disintegration, the content of the PA tablet was spread over a larger area of the gastric mucosa (1088 mm²) than was observed with the P tablets (408 mm²). Three of the 8 volunteers taking the P tablet developed hyperaemia, interstitial bleeding or erosions of the mucosa of the stomach. No such reactions were seen with the PA tablets. Serum concentrations of mecillinam following ingestion of pivmecillinam tablets were determined in three groups of subjects; fasting volunteers, both supine and ambulant, and in ambulant subjects who took the preparation with a light meal. There was a tendency for the new PA tablets to produce a higher peak serum level as well as greater bioavailability of mecillinam. Administration of the PA tablets with a meal significantly increased the peak serum level and total bioavailability of the drug. On the basis of our observations we recommend adoption of the new PA tablet, because of its quick passage through the oesophagus and its more rapid and complete disintegration in the stomach.     

Pharmacokinetics and relative bioavailability of a new chewable, buffered acetylsalicylic acid tablet formulation in comparison to a conventional plain tablet.

The pharmacokinetics of acetylsalicylic acid (ASA) and its main metabolite salicylic acid (SA) following single dose administration of a new chewable, buffered ASA tablet formulation and a conventional plain ASA tablet formulation were investigated in 12 healthy male subjects. The volunteers received in a randomized, crossover design two pharmaceutical units of both formulations containing 500 mg ASA each after an overnight fast on an empty stomach. ASA and SA in the collected plasma and urine samples were determined using an internally standardized validated HPLC method. Regarding the normalized extent parameters for ASA, an increase of about 114% for the maximum concentration (Cmax,norm) and about 16% for the area under the curve (AUC0----infinity,norm) was found for the new chewable, buffered tablet formulation as compared to the plain tablet. Comparing the corresponding parameters for the main metabolite, both formulations were statistically equivalent. The quotient of normalized areas (QAUC0-20min, norm/AUC0----infinity,norm) for ASA was higher by about 124% for the new formulation, indicating an increased and faster absorption during the first 20 min after administration. The time of the concentration maximum did not differ statistically. These data indicate that the new chewable, buffered ASA tablet formulation shows a significant benefit as compared to the plain ASA tablet. The new tablet produced higher plasma ASA concentrations in a shorter time, which is clinically important since higher ASA concentrations are assumed to be related to an improved analgesic efficacy.     

Synthesis,characterization, biological evaluation,and molecular docking studies of some piperonyl-based 4-thiazolidinone derivatives

Heterocyclic compounds are of particular importance among pharmacologically active compounds. In this study, some piperonyl-based 4-thiazolidinone derivatives ( 2a–i ) were synthesized and characterized by spectroscopic assays. All molecules were tested as enzyme inhibitory factors. These compounds were effective inhibitors of the enzymes acetylcholinesterase (AChE), α-glycosidase (α-Gly), and the human carbonic anhydrase I and II isoforms (hCA I and II), with K_i values in the range of 8.90–66.51 nM for α-Gly, 94.8–289.5 nM for hCA I, 106.3–304.6 nM for hCA II, and 0.55–2.36 nM for AChE. The synthesized molecules were also studied theoretically. Molecular docking calculations were performed to investigate the interaction between the target protein and molecules. CA inhibitor compounds have been clinically used for almost 60 years as antiglaucoma and diuretic drugs. The inhibition of the AChE enzyme results in the blockage of ACh hydrolysis. On the contrary, the design of inhibitor compounds or/and modulators for AChE is of major interest as it is one of the most popular tools to prevent Alzheimer's disease.     

Development of a new cyclosporine formulation based on poly(caprolactone) microspheres

The present study describes the development of a new cyclosporine formulation based on polycaprolactone (PCL) microspheres (MS) prepared by the solvent evaporation method. Ternary phase diagrams were used to identify the domains where MS were formed. The application of central composite designs established the influence of several technological (stirring speed) and formulation factors (polymer and surfactant amounts, and organic solvent volume) on the size of PCL MS. Cyclosporine-loaded MS of a size around 2.5 #181;m were prepared and characterized. The stability of the systems, either alone or loaded with cyclosporine, stored at 8#176;C and room temperature (RT) was assessed as well. Freeze-drying was evaluated as an alternative method to achieve longterm stability. The experimental design showed that the stirring speed and the organic phase volume were the only parameters significantly affecting the MS size. Experimental conditions selected to obtain CyA-loaded MS of 2.5 #181;m resulted in a high entrapment percentage (98.4 - 0.66%), with the drug dissolved or molecularly dispersed within the dense polymeric matrix of MS. After 12 months of storage at 8#176;C and RT, PCL MS remained physically stable, although the crystallinity of the polymer increased by 35% upon storage at both temperatures. Freeze-drying studies revealed that MS could be successfully lyophilized in the absence of cryoprotectants without significant changes of the drug entrapment; however, the presence of at least 5% cryoprotectant was essential to keep the initial particle size. Therefore, a stable MS-based CyA formulation was easily prepared and characterized. This formulation offer the possibility of CyA administration through different routes.     

Preparation and evaluation of an orally fast disintegrating tablet formulation containing a hydrophobic drug

Abstract

Orally fast disintegrating tablets (FDTs or ODTs) have received ever-increasing demand during the last decade, and the field has become a rapidly growing area in the pharmaceutical industry. Upon introduction into the mouth, these tablets dissolve or disintegrate in the mouth in the absence of additional water for easy administration of active pharmaceutical ingredients. Although the FDT area has passed its infancy, as shown by a large number of commercial products on the market, there are still many aspects to improve in the FDT formulations. Despite advances in the FDT technologies, formulation of hydrophobic drugs is still a challenge, especially when the amount of drug is high. In this study, a new solution is being developed to incorporate higher doses of a model hydrophobic drug; meloxicam, without affecting the fast disintegrating properties of the formulation. In order to enhance the solubilization of meloxicam in FDT formulations, β cyclodextrin inclusion complex of the drug is prepared and FDTs containing meloxicam--β cyclodextrin inclusion complex (F1 A and F2 A) were compared and evaluated with the FDTs containing pure meloxicam (F1 and F2) by means of in vitro quality control tests.     

An alternative approach for the safety evaluation of new and existing chemicals, an exercise in integrated testing

Various in vitro and in silico methods without animals were applied to 10 substances listed on ELINCS with a complete VIIA base-set available at NOTOX. The hazard assessment for these substances was performed on basis of available non-animal data, QSAR, PBBK-modelling and additional, new in vitro testing was applied. Based on these data predictions on fish toxicity, acute toxicity, skin- and eye-irritation, sensitisation, and toxicity after repeated dosing were made. The predictions were compared with the outcome of the in vivo tests. Nine out of ten predictions on fish LC(50) proved to be correct. For skin- and eye-irritation 70% was predicted correctly. Sensitisation was predicted correctly for 7 out of 10 substances, but three false negatives were found. Acute oral toxicity (LD(50)) and repeated dose toxicity were less successful (5 out of 10 and 2 out of 10 correct predictions, respectively); application of the PBBK model proved successful. Acute dermal toxicity was predicted correctly in 9 out of 10 cases. In general an over-estimation of systemic toxicity was found, which can be explained by an over-prediction of cytotoxicity and worst case assumptions on absorption and binding to (plasma) proteins. This integrated approach leads to a 38% reduction of laboratory animals.     

Toxicological evaluation of ammonium 4,8-dioxa-3H-perfluorononanoate,a new emulsifier to replace ammonium perfluorooctanoate in fluoropolymer manufacturing

Ammonium 4,8-dioxa-3H-perfluorononanoate (ADONA) was developed to replace ammonium perfluorooctanoate (APFO) as an emulsifier in the manufacture of fluoropolymers. The toxicity of ADONA was evaluated in acute and repeat-dose studies of up to 90-days duration, and in eye and skin irritation, dermal sensitization, genotoxicity, and developmental toxicity studies. ADONA was also evaluated as a peroxisome proliferator-activated receptor alpha (PPARα) agonist in rats. ADONA was moderately toxic orally and practically non-toxic dermally in acute studies in rats. It was a mild skin irritant and a moderate to severe eye irritant in rabbits. It was a weak dermal sensitizer in local lymph node assays in mice. ADONA was not genotoxic based on the weight of evidence from five assays. It was not developmentally toxic in rats except at maternally toxic doses. ADONA was a possible PPARα agonist in male rats. The liver was the primary target organ in male rats and the kidney was the primary target organ in female rats. NOAELs in 28- and 90-day oral studies in rats were 10 mg/kg/day for males and 100 mg/kg/day for females. These findings demonstrate that the toxicity profile for ADONA is acceptable for its intended use and is superior to that of APFO.     

Development of a new cyclosporine formulation based on poly(caprolactone) microspheres.

The present study describes the development of a new cyclosporine formulation based on polycaprolactone (PCL) microspheres (MS) prepared by the solvent evaporation method. Ternary phase diagrams were used to identify the domains where MS were formed. The application of central composite designs established the influence of several technological (stirring speed) and formulation factors (polymer and surfactant amounts, and organic solvent volume) on the size of PCL MS. Cyclosporine-loaded MS of a size around 2.5 microm were prepared and characterized. The stability of the systems, either alone or loaded with cyclosporine, stored at 8 degrees C and room temperature (RT) was assessed as well. Freeze-drying was evaluated as an alternative method to achieve long-term stability. The experimental design showed that the stirring speed and the organic phase volume were the only parameters significantly affecting the MS size. Experimental conditions selected to obtain CyA-loaded MS of 2.5 microm resulted in a high entrapment percentage (98.4 +/- 0.66%) with the drug dissolved or molecularly dispersed within the dense polymeric matrix of MS. After 12 months of storage at 8 degrees C and RT, PCL MS remained physically stable, although the crystallinity of the polymer increased by 35% upon storage at both temperatures. Freeze-drying studies revealed that MS could be successfully lyophilized in the absence of cryoprotectants without significant changes of the drug entrapment; however, the presence of at least 5% cryoprotectant was essential to keep the initial particle size. Therefore, a stable MS-based CyA formulation was easily prepared and characterized. This formulation offer the possibility of CyA administration through different routes.