Assessment of hypolipidemic, anti-inflammatory and antioxidant properties of medicinal plant Erica multiflora in triton WR-1339-induced hyperlipidemia and liver function repair in rats: A comparison with fenofibrate.

Hyperlipidemia is a serious health threat that has been linked to oxidative stress and systemic inflammation, causing among many other disorders essentially liver disease. The current study was conducted to evaluate the antihyperlipidemic, antioxidant and anti-inflammatory potential of methanol leaf extract from Erica multiflora (M-EML). Triton WR-1339-induced hyperlipidemic rats were divided into six groups: control group (CG), hyperlipidemic group (300 mg/kg body weight "BW") (HG), hyperlipidemic group treated with M-EML (150 and 250 mg/kg) (HG + M-EML), normal rats treated with M-EML (250 mg/kg) and fenofibrate-treated group (HG + FF) (65 mg/kg). After 24 h of administration, triton WR-1339 induced a significant increase in lipid profile, atherogenic index (AI) and Coronary Risk Index (CRI) in HG group compared to control group. Furthermore, triton WR-1339 administration induced alteration in the status of pro-inflammatory markers (aspartate transaminase, alanine transaminase, IFN-γ and Nitric oxide production). HG group showed also, a high level of lipid peroxidation, an altered antioxidant enzyme profiles and an increase in DNA damages, in liver. However, orally administration of M-EML mitigates significantly these disorders, proving hence a protective potential against triton WR-1339-induced hyperlipidemia. These findings suggest that M-EML extract could be used as functional foods and natural adjuvant treatment of hyperlipidemia.

Assessment of the Antioxidant/Hypolipidemic Relationship of Sideritis hyssopifolia in an Experimental Animal Model.

Many publications have described the potential cardioprotective action of different medicinal plants, relating this effect with blood lipid levels. However, these publications do not justify the right amount of plant administered, which can vary greatly. Sideritis hyssopifolia is a little woody plant endemic to western and southwestern Europe. We have quantified its antioxidant activity, which can be used as an indicator of its cardioprotective action. This study evaluates the antioxidant capacity of Sideritis hyssopifolia to design a feed whose hypolipidemic effects are proven in cholesterol-fed New Zealand rabbits. Antioxidant action was assessed in infusions, which were prepared with 1 or 3 g of plant in 200 mL of water by using an ABTS assay and expressed as Ascorbic acid Equivalent Antioxidant Capacity (AEAC). Aqueous infusions with infusion times of 10 min and prepared with 3 g plant exhibited the strongest antioxidant activity. Sideritis hyssopifolia showed an intermediate antioxidant capacity for the concentrations and times of the infusion tested. According to our results, we suggest incorporating 2.36 g of S. hyssopifolia every 150 g of rabbit feeding stuff (15.73 g/kg). This chow decreased cholesterol, HDL-cholesterol, LDL-cholesterol, and triglycerides levels in cholesterol-fed rabbits, as well as the atherogenic index. This reduction was similar to that obtained with simvastatin.

Variability in Potency Among Commercial Preparations of Berberine.

Berberine is an isoquinoline alkaloid plant extract that is widely available as a dietary supplement in the United States and has demonstrated efficacy in the treatment of type 2 diabetes mellitus and dyslipidemia. Because of its increased use and purported pharmacological properties, potential variations in product quality could pose a barrier to berberine's safety and effectiveness in clinical practice. Thus, this study evaluated the potency of dietary supplements containing berberine available in the U.S. commercial market. Fifteen unique dietary supplements containing berberine were purchased through U.S. dietary supplement vendors. For each product, berberine was extracted from 3 unique capsules and analyzed by ultra-high-performance liquid chromatography tandem mass spectrometry. Percentage content based on the product label claim was determined for each product. The average berberine content across the products was found to be 75% ± 25% of the product label claim, with product potency ranging from 33% to 100%. Nine of the 15 tested products (60%) failed to meet the potency standards of 90% to 110% of labeled content claim, as commonly required of pharmaceutical preparations by the U.S. Pharmacopeial Convention. Evaluation of the relationship between product cost and the measured potency failed to demonstrate an association between quality and cost. Variability in product quality may significantly contribute to inconsistencies in the safety and effectiveness of berberine. In addition, the quality of the berberine product cannot be inferred from its cost.

Experimentally designed lyophilized dry emulsion tablets for enhancing the antihyperlipidemic activity of atorvastatin calcium: Preparation, in-vitro evaluation and in-vivo assessment.

This article presents the development of lyophilized orally disintegrating tablets prepared with the dry emulsion technique to enhance the in-vitro dissolution and in-vivo performance of the poorly bioavailable drug atorvastatin calcium (ATV). Tablets were fabricated by freeze-drying o/w emulsions of ATV. The Emulsions were prepared using a matrix former solution (alginate or gelatin, 2 or 4%) containing a sugar alcohol (mannitol) and a collapse protectant (glycine) as the water phase and Labrafac® as the oil phase in the presence of surfactant (synperonic® PE/P 84 or synperonic® F108) under proper homogenization. The influence of formulation parameters on friability of the prepared tablets, disintegration time and in-vitro dissolution of the drug from these tablets were investigated. Results showed the significant influence of the matrix former and emulsifier type on the disintegration time. In-vitro dissolution study revealed the enhanced dissolution rate of ATV from the lyophilized dry emulsion tablets (LDET) compared to the plain drug. DSC and XRD studies of the optimized ATV-loaded LDET proved the presence of the drug in the amorphous form. SEM images showed the intact, porous and non-collapsible structure of the prepared LDET with complete loss of ATV crystallinity. Administration of ATV-loaded LDET to high fat diet-induced hyperlipidemic rats demonstrated a significant decrease (p<0.05) in the serum and tissue levels of the tested parameters compared to the market product used. The obtained results suggest a promising, easy-to-manufacture and effective dosage form for the treatment of hyperlipidemia.

In Vitro Assessment of CYP-Mediated Drug Interactions for Kinsenoside, an Antihyperlipidemic Candidate.

Kinsenoside, the herb-derived medicine isolated from the plant Anoect chilus, has diverse pharmacological actions, and it is considered to be a promising antihyperlipidemic drug candidate. This study evaluates the effects of kinsenoside on CYP enzyme-mediated drug metabolism in order to predict the potential for kinsenoside-drug interactions. Kinsenoside was tested at different concentrations of 0.1, 0.3, 1, 3, 10, 30, and 100 µM in human liver microsomes. The c Cktail probe assay based on liquid chromatography-tandem mass spectrometry was conducted to measure the CYP inhibitory effect of kinsenoside. Subsequently, the metabolism profiles of amlodipine and lovastatin in human liver microsomes were analyzed following co-incubation with kinsenoside. The concentration levels of the parent drug and the major metabolites were compared with the kinsenoside-cotreated samples. The effect of kinsenoside was negligible on the enzyme activity of all the CYP isozymes tested even though CYP2A6 was slightly inhibited at higher concentrations. The drug-drug interaction assay also showed that the concomitant use of kinsenoside has a non-significant effect on the concentration of lovastatin or amlodipine, and their major metabolites. So, it was concluded that there is almost no risk of drug interaction between kinsenoside and CYP drug substrates via CYP inhibition.

Assessment of simvastatin niosomes for pediatric transdermal drug delivery.

The prevalence of childhood dyslipidemia increases and is considered as an important risk factor for the incidence of cardiovascular disease in the adulthood. To improve dosing accuracy and facilitate the determination of dosing regimens in function of the body weight, the proposed study aims at preparing transdermal niosomal gels of simvastatin as possible transdermal drug delivery system for pediatric applications. Twelve formulations were prepared to screen the influence of formulation and processing variables on critical niosomal characteristics. Nano-sized niosomes with 0.31 µm number-weighted size displayed highest simvastatin release rate with 8.5% entrapment capacity. The niosomal surface coverage by negative charges was calculated according to Langmuir isotherm with n = 0.42 to suggest that the surface association was site-independent, probably producing surface rearrangements. Hypolipidemic activities after transdermal administration of niosomal gels to rats showed significant reduction in cholesterol and triglyceride levels while increasing plasma high-density lipoproteins concentration. Bioavailability estimation in rats revealed an augmentation in simvastatin bioavailability by 3.35 and 2.9 folds from formulation F3 and F10, respectively, compared with oral drug suspension. Hence, this transdermal simvastatin niosomes not only exhibited remarkable potential to enhance its bioavailability and hypolipidemic activity but also considered a promising pediatric antihyperlipidemic formulation.

Gastrointestinal behavior of nano- and microsized fenofibrate: In vivo evaluation in man and in vitro simulation by assessment of the permeation potential.

INTRODUCTION: The purpose of this study was (i) to evaluate the gastrointestinal behavior of micro- and nanosized fenofibrate in humans and (ii) to develop a simple yet qualitatively predictive in vitro setup that simulates the observed absorption-determining factors. MATERIALS AND METHODS: Commercially available micro- and nanoparticles of fenofibrate (Lipanthyl® and Lipanthylnano®, respectively) were administered orally to five healthy volunteers in fasting and postprandial conditions. Intraluminal and systemic drug concentrations were determined as reference data for the development of a predictive in vitro setup. To capture the observed solubility/permeability interplay, in vitro dissolution testing was performed in the presence of a permeation bag with sink conditions. RESULTS: In fasting conditions, intake of nanosized fenofibrate generated increased duodenal concentrations compared to microsized fenofibrate, which was reflected in an improved systemic exposure. In postprandial conditions, duodenal concentrations were greatly enhanced for both formulations, however without an accompanying increase in systemic exposure. It appeared that micellar encapsulation of the highly lipohilic fenofibrate limited its potential to permeate from fed state intestinal fluids. To capture these in vivo observations in an in vitro setup, classic dissolution testing was combined with permeation assessment into a permeation bag with sink conditions. In case of fasting conditions, the dissolution/permeation approach allowed for an improved discriminative power between micro- and nanosized fenofibrate by better simulating the dynamic interplay of dissolution and absorption. In case of postprandial conditions, the observed solubility-permeability interplay could be simulated using the dissolution/permeation approach in combination with biorelevant media (FeSSGFFortimel and FeSSIF-V2) to mimic micellar entrapment and reduced permeation potential of fenofibrate. CONCLUSION: For the first time, reduced permeation of a lipophilic drug despite increased intraluminal concentrations, was demonstrated in humans. Dissolution testing using biorelevant media in combination with permeation assessment into a sink permeation bag appeared to be a simple yet pragmatic approach to capture this solubility-permeability interplay in early formulation evaluation.

Bio-generation of stable isotope-labeled internal standards for absolute and relative quantitation of phase II drug metabolites in plasma samples using LC-MS/MS.

Quantification of drug metabolites in biological samples has been of great interest in current pharmaceutical research, since metabolite concentrations and pharmacokinetics can contribute to a better understanding of the toxicity of drug candidates. Two major categories of Phase II metabolites, glucuronide conjugates and glutathione conjugates, may cause significant drug toxicity and therefore require close monitoring at early stages of drug development. In order to achieve high precision, accuracy, and robustness, stable isotope-labeled (SIL) internal standards (IS) are widely used in quantitative bioanalytical methods using liquid chromatography and tandem mass spectrometry (LC-MS/MS), due to their capability of compensating for matrix effects, extraction variations and instrument response fluctuations. However, chemical synthesis of SIL analogues of Phase II metabolites can often be very difficult and require extensive exploratory research, leading to higher cost and significant delays in drug research and development. To overcome these challenges, we have developed a generic method which can synthesize SIL analogues of Phase II metabolites from more available SIL parent drugs or SIL conjugation co-factors, using in vitro biotransformation. This methodology was successfully applied to the bio-generation of SIL glucuronide conjugates and glutathione conjugates. The method demonstrated satisfactory performance in both absolute quantitation and assessment of relative exposure coverage across species in safety tests of drug metabolites (MIST). This generic technique can be utilized as an alternative to chemical synthesis and potentially save time and cost for drug research and development.

Assessment of the genotoxic activity of alpha-asarone and its derivatives in the comet assay.

In our previous paper we examined mutagenic and genotoxic activity of 4 alpha-asarone isomers 2-5 exhibiting relatively high hypolipidemic activity. In the present paper, we examined genotoxic activity of alpha-asarone and its isomers as the ability to damage cellular DNA, evaluated in the comet assay. Additionally, mutagenic activity of alpha-asarone in Ames test has been examined. The Ames test for alpha-asarone was carried out in accordance with the guidelines of the PN-EN ISO 10993-3 standard. Compounds 4 and 5 were found to be devoid of any genotoxic activity while maintaining their hypolipemic potential. Because mutagenic activity of compound 4 was also minor it could be considered as a candidate for further pharmacological evaluation. Genotoxic but not mutagenic activity of alpha-asarone has been confirmed.

The occurrence and bioactivity of polyphenols in Tunisian olive products and by-products: a review.

Polyphenols have become a subject of intense research because of their perceived beneficial effects on health due to their anticarcinogenic, antiatherogenic, anti-inflammatory, and antimicrobial activities. It is well known that olives and their derivatives are rich in phenolic substances with pharmaceutical properties, some of which exert important antioxidant effects. The characterization and quantification of their polyphenol composition is one of the first steps to be taken in any evaluation of the putative contribution of the olive to human health. This review is concerned with polyphenols in Tunisian olive (Olea europaea L.) products (fruit and oil) and some by-products (leaves and olive-mill wastewater) with an emphasis on the analytical methods used, as well as the biological activities described in recent years.

The assessment of human regional drug absorption of free acid and sodium salt forms of acipimox, in healthy volunteers, to direct modified release formulation strategy.

Acipimox is an analog of nicotinic acid and is indicated for the treatment of dyslipidemia. It is also believed to improve glucose control by enhancing insulin sensitivity. The purpose of this study was to direct modified release (MR) formulation strategy by comparing the bioavailability of two forms of acipimox (free acid and sodium salt) from the distal small bowel (DSB) and colon with an immediate release formulation. Two parallel groups of healthy volunteers completed an open label, non-randomized, three-way crossover study. The rate and extent of acipimox absorption was highest following administration of the immediate release capsules, and was not influenced by the form of the drug administered. Following administration to the DSB, the relative bioavailability was approximately 52% and 30% for the salt form and free acid form, respectively. Following administration to the colon, the extent of absorption was further reduced. The data indicate that bioavailability from the DSB was limited by the solubility of the drug coupled with an absorption window, whilst absorption from the colon was limited by permeability. The study provided detailed information to support and guide the formulation strategy for a MR form of acipimox, which may improve the treatment of adult patients with type II diabetes and dyslipidemia.

Chemical characterization of the lipophilic fraction of Livistona decipiens and Livistona chinensis fruit pulps (Palmae) and assessment of their anti-hyperlipidemic and anti-ulcer activities.

The oil of the dried pulps of Livistona decipiens and L. chinensis palm fruits have been studied for the first time by gas chromatography-mass spectrometry for their unsaponifiable matter (USM) and fatty acid composition (FAME). The antihyperlipidemic and anti-ulcer activities for both oils were also assayed. The principal fatty acid of L. decipiens pulp oil was oleic acid (53.4 %) and of L. chinensis pulp oil palmitic acid (47.4 %). In relation to anti-hyperlipidemic properties, the pulp oil of L. decipiens presented a better profile than that of L. chinensis, in comparison with the reference standard (simvastatin). In addition, both pulp oils showed high anti-ulcer activity using an indomethacin-induced ulceration technique in rat stomach. The relationship between the anti-hyperlipidemic, anti-ulcer and chemical composition of the pulp oils is also discussed.

Preformulative assessment of preformed complexes of gemfibrozil, with cyclodextrins.

The aim of this study is to carry out preformulative investigations on preformed inclusion complexes of the poorly water-soluble, lipid-lowering agent gemfibrozil and naturally occurring cyclodextrins (CDs). Phase solubility studies showed a linear AL- type diagram with alpha, beta, and y cyclodextrins, indicating the formation of inclusion complexes in a 1:1 molar ratio with all the three CDs. beta-CD-gemfibrozil complex having a maximum stability constant of 148.88 M(-1) was selected for preparation of preformed inclusion complex by kneading, co-precipitation, co-evaporation, and freeze-drying and compared with the physical mixture. The kneaded product was subjected to microwave-drying, with this mode of drying studied as an alternative method for preparation of the complex. The prepared complexes were assessed by equilibrium solubility analysis and intrinsic dissolution rate studies. Further characterization was done by differential scanning calorimetry, X-ray powder diffractometry, Fourier transform infrared spectroscopy, and scanning electron microscopy. The freeze-dried product was identified as the inclusion complex having the maximum intrinsic dissolution rate and hence was assessed for changes in permeability characteristics. pH partition studies and partial in vivo permeability studies showed no changes in the permeability of the freeze-dried product when compared to the pure drug.

Assessment of hypolipidaemic activity of three thiazolidin-4-ones in mice given high-fat diet and fructose.

Three 4-thiazolidinones, two with nicotinamide (NAT1 and NAT2) and one with 4-chlorophenoxyacetamide (PAT1) side chains were evaluated for their hypolipidaemic, hypoglycaemic activity in Swiss albino mice fed a high-fat diet along with fructose administered in drinking water. NAT1 and PAT caused reduction of elevated triglycerides, cholesterol and glucose; NAT2 was effective only against triglycerides. Nicotinamide side chain might have contributed to the lipid lowering effect of both NAT1 and NAT2, but the bulky group of the latter could have affected proper binding to the receptor sites, making it ineffective against elevated cholesterol. On the other hand, the 4-chlorophenoxyacetamide side chain of PAT might have exerted powerful hypolipidaemic activity, despite the bulky substitution at C2. As antioxidants, NAT2 and PAT1 showed superior activity, compared to NAT1. The thiazolidinone ring might be responsible for the lipid lowering effect, which is however, modified by the type of substitutions at C2 and N of the ring. Detailed study is warranted to explain the mechanism of action of these compounds as also to make more potent ones.

Bezafibrate removal by means of ozonation: primary intermediates, kinetics, and toxicity assessment.

Bezafibrate (BZF) is a lipid regulator largely used for the treatment of hyperlipidaemia. As a result of its wide use, unmetabolized BZF is released in the environment with potential toxic effects for aquatic living organisms. The results obtained in this work show that ozonation is an efficient method to degrade BZF: after 10 min of treatment (corresponding to a dose of 0.73 mmol L(-1) of ozone), the complete BZF abatement is achieved, starting from an initial concentration of 0.5 mmol L(-1). However, only a small part of the substrate is mineralized. Two different experimental approaches (absolute and competition method) are adopted to estimate the second-order kinetic constants for the ozone attack at pH=6.0, 7.0 and 8.0. A good agreement was observed between the two kinetic methods adopted. The identification of main intermediates, attempted by high-performance liquid chromatograph (HPLC)-MS technique, indicates that the oxidation of BZF develops through both the hydroxylation of the aromatic ring and the attack of ozone on the unchlorinated aromatic one. The assessment of by-products biodegradability and acute toxicity demonstrates that ozonation is a suitable technique to improve the biodegradability and reduce the toxicity of waters containing BZF.

[Emergy analysis on an added loop in Spartina alterniflora ecological engineering].

Based on the principle of multi-grade utilization of resources to get ecological, economic and social benefits of ecological engineering, this paper designed an added loop, following the Spartina alterniflora Ecological Engineering (SAEE). All the added loop design included SAEE, and the capsule was named SAEEC. In the added loop design, the Biological Mineral Liquid (BML) was made into antihyperlipidemia capsules, of which, the total flavonoids added up to 9.58 mg x g(-1). Emergy analysis method was applied to evaluate the SAEEC project. Compared with SAEE, the added loop design increased emergy investment ratio (EIR) by 1.37 fold, net economic benefit of the SAEEC by 2.13 fold, economic yield/input ratio by 1.46 fold, net emergy yield (NEY) by 3.18 fold, and net emergy yield ratio (EYR) by 2.20 fold, showing its more efficiency.