Pomegranate extract-loaded sphingosomes for the treatment of cancer: Phytochemical investigations, formulation, and antitumor activity evaluation.

AIM: Formulation of Pomegranate Extracts (PE)-loaded sphingosomes as an antitumor therapy for the intravenous and passive targeted delivery to various tumor types, especially that of the breast, colon, and uterus; to increase the therapeutic activity and decrease the adverse effects profile. METHODS: The pericarp and seeds' juice of Punica granatum were each extracted using D.W. and ethanol. Phytochemical investigation of all extracts was carried out including total phenolics, flavonoids, and anthocyanins contents, the antioxidant activity, as well as HPLC analysis of phenolics and flavonoids. The antitumor potential of all extracts was also tested utilizing three cell lines: MCF-7, HeLa, and HCT116. The candidate extract was chosen for the formulation phase and was entrapped into the sphingosomes using the thin-film hydration method and employing three different PE: lipids weight ratios. The synthesized formulations were characterized for their size, morphological features, zeta potential, entrapment efficiency, and in vitro drug release and kinetics modeling studies. The optimized formula was further analyzed by FTIR spectroscopy and electron microscopy. The antitumor activity of F2 was also investigated using the same cancer cell lines compared to the plant extract. RESULTS: The highest phenolics, flavonoids, and anthocyanins contents were observed in the ethanolic pericarps extract (EPE), followed by the ethanolic seeds extract (ESE). Consequently, EPE showed a higher antitumor activity hence it was selected for the formulation phase. PE-loaded sphingosomes formula (F2) was selected for having the highest EE% (71.64%), and a sustained release profile with the highest in vitro release (42.5±9.44%). By employing the DDSolver, the Weibull model was found the most suitable to describe the PE release kinetics compared to other models. The release mechanism was found to follow Fickian diffusion. Simulated pharmacokinetic parameters have portrayed F2 as the candidate formula, with the highest AUC (536.095) and slowest MDT (0.642 h). In addition, F2 exhibited a significant (p>0.05) stronger and prolonged anticancer effect against MCF-7, HeLa, and HCT116 cell lines at all concentrations tested compared to the free extract. CONCLUSION: The results proved that sphingosomes are an effective delivery system, improving pharmacological efficacy and reducing serious side effects of anticancer medications and natural products.

Preparation and antihepatotoxicity activity of Fagonia indica extract and its solid dispersion formulation.

This study aimed to evaluate and compare the antihepatotoxicity effect of Fagonia indica extract and its solid dispersion formulation (SD) against paracetamol-induced hepatotoxicity in rats. Dried Ethanolic plant extract was prepared by cold maceration in ethanol followed by solvent evaporation under reduced pressure. Quality control of crude extract was performed and the total phenolic and flavonoid contents were determined. Solid dispersion (SD) formulations were prepared by solvent evaporation technique and optimized with respect to drug solubility. Antihepatotoxicity activities of Fagonia indica extract and optimized solid dispersion were performed against paracetamol-induced hepatotoxicity in rats. Quality control parameters like total ash, acid insoluble ash, water soluble ash, crude fiber content and moisture content were within the acceptable limits. Total flavonoid and phenolic contents were found to be 31.289mg quercetin equivalents/g and 40.28mg gallic acid equivalent/g respectively. TLC Investigation of the plant extract revealed the presence of gallic acid, kaempferol and quarcetin. Optimized SD formulation with 200 mg of the dried extract, 350mg of PEG 4000 and 50mg of Tween 20 showed almost four-fold increasing in the solubility of the extract in water. The average hydrodynamic diameter of extract particles was reduced from 1972 nm to 437.6nm when prepared as SD. SD formulation showed highest antihepatotoxicity activity compared with plain plant extract at the same concentration. Optimized SD formulation at 500mg dose showed complete recovery from hepatotoxicity induced by paracetamol in rats. Therefore, SD is found to be one of the promising strategy to enhance the antihepatoxicity activity of Fagonia indica plant.

Therapeutic potentials of Crataegus azarolus var. eu- azarolus Maire leaves and its isolated compounds.

BACKGROUND: Hyperglycemia is a complicated condition accompanied with high incidence of infection and dyslipidemia. This study aimed to explore the phyto-constituents of Crataegus azarolus var. eu- azarolus Maire leaves, and to evaluate the therapeutic potentials particularly antimicrobial, antihyperglycemic and antihyperlipidemic of the extract and the isolated compound (3ß-O-acetyl ursolic acid). METHODS: Total phenolics and flavonoidal contents were measured by RP-HPLC analysis. Free radicals scavenging activity of different extraction solvents was tested in-vitro on DPPH free radicals. The antimicrobial activity of the ethanolic extract and its fractions as well as the isolated compounds were evaluated in-vitro on variable microorganisms. Animal models were used to evaluate the antihyperglycemic and antihyperlipidemic activities of the ethanolic extract along with the isolated compound (3ß-O acetyl ursolic acid). RESULTS: RP- HPLC analysis of the phenolics revealed high content of rutin, salicylic and ellagic acids. Six compounds belonging to triterpenes and phenolics were isolated from chloroform and n-butanol fractions namely: ursolic acid, 3ß-O-acetyl ursolic acid, ellagic acid, quercetin 3-O-ß methyl ether, rutin and apigenin7-O-rutinoside. Ethanolic extract showed the highest DPPH radical scavenger activity compared to other solvents. Ethanolic extract, hexane fraction, ursolic acid, 3ß-O acetyl ursolic acid and quercetin 3-O-methyl ether showed variable antimicrobial activity against E. coli, P. aeruginosa, S. aureus, and C. albicans. Administration of the ethanolic extract or 3ß-O acetyl ursolic acid orally to the mice reduced blood glucose significantly in a time- and dose-dependent manner. Ethanolic extract significantly reduced LDL-C, VLDL-C, TC and TG and increased HDL-C in rats. Ethanolic extract and 3ß-O acetyl ursolic acid reduced in-vitro activity of pancreatic lipase. CONCLUSION: This study reveals that Crataegus azarolus var. eu- azarolus Maire has the efficiency to control hyperglycemia with its associated complications. This study is the first to evaluate antihyperglycemic and antihyperlipidemic potentials of 3ß-O acetyl ursolic acid.

Impact of phenolic composition on hepatoprotective and antioxidant effects of four desert medicinal plants.

BACKGROUND: Flavonoids and other polyphenols play a protective role in liver diseases and possess a high antioxidant capacity. OBJECTIVE: To compare and evaluate the antioxidant and hepatotoprotective activities of 4 deserts plants, Fagonia indica Burm. f., Calotropis procera R.Br., Zygophylum hamiense Schweinf. and Salsola imbricata Forssk. in correlation to their composition especially their phenolic content. METHODS: The influence of extracting solvent on total phenolic and flavonoidal contents was assessed spectrophotometrically. The flavonoid and other polyphenolic components of the methanol extracts were analyzed by RP-HPLC. DPPH radical scavenging potential of the different extracts was estimated. The hepatoprotective and antioxidant activities of the extracts against CCl4-induced hepatotoxicity in mice were evaluated. RESULTS: The flavonol quercitrin and rosmarinic acid were major in the F. indica, C. procera and S. imbricata samples, while rutin prevailed in that of Z. hamiense. The ethanolic and methanolic extracts showed noticeable DPPH radical-scavenging activity as compared to ascorbic acid. Assessment of liver enzymes revealed that oral administration of the extracts did not show any evidence of hepatotoxicity. Moreover, protection against CCl4-induced liver damage was evident upon administration of three plants extracts namely, F. indica, C. procera and S. imbricata. CONCLUSION: Overall, hepatotoxicity induced by CCl4 was effectively prevented by the three plants extracts through scavenging of free radicals and by boosting the antioxidant capacity of the liver. The protective effect of the plants could be attributed to their high quercitrin and rosmarinic acid contents.

A new natural gel of Fagonia indica Burm f. extract for the treatment of burn on rats.

Fagonia indica Burm f. (Mushikka or white spine) is a plant distributed in the deserts of Asia and Africa and reported to be medicinal in the scientific literature as well as in the folk medicine. Earlier investigations, the authors isolated a number of bioactive constituents from the plant including flavonoids, sterols and tritepenoids; In addition its flavonoidal content was found remarkably high reaching 3% (calculated as flavonol on dry weight). The present study is an attempt to formulate, characterize and evaluate a natural wound-healing gel preparation containing the crude plant extract. Three formulae (F1-F3) were prepared. The gel properties such as viscosity, swelling ratio, bio-adhesion, in vitro release, stability, microbiological studies, in vivo burn healing test on rats and histopathological features were assessed. The results of the in vitro evaluation and stability studies showed that F3 (0.5% (w/w) of plant extract in 4% (w/w) chitosan) was significantly (p<0.05) the superior compared to other formulations. Besides, from the in vivo burn healing and histological results, F3 enhanced the skin wound re-epithelialization and speed up the healing process compared to the conventional commercial product. Thus, the Fagonia extract loaded chitosan topical gel would be used successfully in burn wound care.

Phenolic Profiling and Evaluation of Contraceptive Effect of the Ethanolic Extract of Salsola imbricata Forssk. in Male Albino Rats.

Reported researches dealing with either composition or bioactivity of Salsola imbricata are limited. This study was conducted aiming to investigate the phenolic composition of the plant and evaluate its efficacy as male contraceptive. Polyphenols, namely, phenolic acids and flavonoids, were qualitatively and quantitatively analysed by RP-HPLC in the hydrolysed methanol extract using two different wavelengths, 280 and 330 nm. The efficiency of different solvents in extracting the plant phenolics was assessed via spectrophotometric determination of the total phenolic and flavonoid contents. Acute toxicity study was carried out on the ethanolic extract to ascertain its safety prior to biological evaluation. The contraceptive effect was assessed, in male rats, by oral administration of the extract at two doses (250 and 500 mg/kg b. wt.), over a period of 65 days. HPLC analyses allowed the identification and quantification of a total of 13 and 8 components in the hydrolysed-methanol extract; the overall phenolic composition was dominated by quercitrin (12.692%) followed by coumaric acid (4.251%). Prolonged oral administration of the ethanolic extract caused slight reduction in the testis weight only. A significant decrease in the sperm count was observed (P < 0.01) in the two treated groups while significant decrease in the epididymal sperm motility was only observed in the high dose group. Morphological abnormalities were observed in sperms of treated animals. No distinct change in serum FSH, LH, and testosterone concentration was recorded. The histopathological findings supported to a high extent these results. The male contraceptive activity of Salsola imbricata could be ascribed to its phenolic components, especially quercitrin.

Anti-inflammatory and gastroprotective activities of the aqueous extract of Micromeria fruticosa (L.) Druce ssp Serpyllifolia in mice.

Micromeria fruticosa is used widely in many Mediterranean regions for various inflammatory conditions. The aim of this work was to assess the anti-inflammatory and gastroprotective activities of the aqueous extract of Micromeria fruticosa. The aqueous extract of Micromeria fruticosa was tested orally in mice at doses of 50, 100 and 200 mg/kg in carrageenan-induced paw edema, vascular permeability, myeloperoxidase activity (MPO) and indomethacin-induced gastric ulceration. In the paw edema model, the extract at dose of 200 mg/ kg, exhibited a significant anti-inflammatory effect, while the extract at 100 and 200 mg/kg reduced significantly the vascular permeability and MPO activity in a dose dependant manner. Oral pretreatment of the aqueous extract reduced significantly the development of gastric lesions induced by indomethacin at dose of 200 mg/kg only. Results suggest that the aqueous extract of Micromeria fruticosa has both anti-inflammatory as well as, gastroprotective activities. Thus it could be used as an alternative or supplementary herbal remedy for the treatment of inflammatory diseases especially when combined with strong anti-inflammatory medications that have ulcerogenic side effects such as NSAIDs.

Constituents and biological activity of the essential oil and the aqueous extract of Micromeria fruticosa (L.) Druce subsp. serpyllifolia.

Micromeria fruticosa L Druce subsp. serpyllifolia is a medicinal herb that is widely used as folk medicine in the treatment of abdominal pains, diarrhea, eye infections, heart disorders, elevated blood pressure, colds and wounds. This study aims to investigation the constituents and biological activity of the essential oil and aqueous extract of the plant that had been collected from Nablus. The oil was prepared by hydro-distillation method and analyzed by GC/MS. The oxygenated constituents were prevalent (87.4%) with the pulegone (58.5%) was the major constituents. Antitumor and analgesic activities of the isolated oil and the aqueous extract of M. fruticosa were investigated. Both the oil and the aqueous extract exhibited marked antitumor activities against Human Colon Tumor cells (HCT) and Mammary Carcinoma F7 (MCF7). The oil showed less IC(50s) against both cell lines (10, 12.7 µg/ml respectively). Also the extract significantly inhibited acetic acid-induced writhing response (p<0.05) and increased hot-plate pain threshold of mice at doses of 100 and 200 mg/kg while the oil did not show any analgesic activity on both models. Therefore, we concluded that the aqueous extract of M. fruticosa has a remarkable inhibitory activity in non-inflammatory reactions as well as inflammatory pain.