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OBJECTIVEThis study investigated cytotoxicity and induction of apoptosis in human cervical cancer cells (HELA) and prostate cancer cells (PC-3) using the most active fraction of Moringa peregrina seed extract.METHODSDried and powdered seeds were extracted using 95% ethanol. The total ethanolic extract was further dissolved in distilled water and separated into petroleum ether, chloroform, ethyl acetate and aqueous extracts. Based on the results of in vitro anticancer studies of all extracts, the most highly active extract was selected for evaluation of apoptosis induction and cell cycle analysis on HELA and PC-3 cells at its half maximal inhibitory concentration using flow cytometry; DNA fragmentation by agarose gel electrophoresis and the expression of protein were measured by Western blot.RESULTSThe chloroform fraction from the ethanolic extract of M. peregrina (CFEE) was the most active antitumor fraction. The selectivity index, determined using the normal Vero cell line, indicated that CFEE had a high degree of selectivity against HELA and PC-3 cells. CFEE induced apoptosis, confirmed by cell cycle arrest at sub-G₀ phase and DNA fragmentation. CFEE induced an increase in mRNA expression of caspase-3, a decrease in Bcl-2 mRNA expression, and decreased ATP levels. CFEE increased protein expression of caspase-3 and decreased protein expression of poly-ADP-ribose polymerase-1 (PARP-1). Flow cytometric analysis showed an appreciable increase in the number of cells in the early apoptotic stage in CFEE-treated HELA and PC-3 cells. CFEE treatment significantly increased lipid peroxidation (malondialdehyde level) in HELA and PC-3 cells.CONCLUSIONSeed extract of M. peregrina displayed a significant antitumor effect through apoptosis induction in HELA and PC-3 cells.

2016-07-25·Chromatography Research International

Development and Validation of New RP-HPLC Method for Simultaneous Determination of Methyl and Propyl Parabens with Levetiracetam in Pure Form and Pharmaceutical Formulation

作者: Abd El-Hay, Soad S. ; Mohram, Mostafa S.

A simple and robust high-performance liquid chromatography (HPLC) method is described for the assay for levetiracetam (LTC), methyl paraben (MHB), and propyl paraben (PHB) either in their pure form or in commercial Levepsy® syrup. The method is selective and stability indicating and all chromatographic conditions were studied to obtain adequate separation of LTC, MHB, and PHB from their degradation products and from excipients. The HPLC separation was carried out on a RP C18 Hypersil BDS analytical column (150 mm × 4.6 mm ID) using gradient elution system. The mobile phase flow rate was 1.5 mLmin−1 and the column temperature was kept at 40°C. Complete separation of the studied components was obtained within a cycle time of 8 min. LTC, MHB, and PHB were eluted at 1.56, 5.86, and 7.85 min, respectively. Detection was carried out at 240 nm using a dual wavelength detector. The method has been validated for linearity, accuracy, precision, specificity, limit of detection, limit of quantitation, robustness, and ruggedness. The proposed method was successfully applied for the determination of LTC in the presence of parabens in Levepsy syrup.

2009-12-01·Int. Journal of Clinical Pharmacology and Therapeutics4区 · 医学

Bioequivalence of two oral formulations of clopidogrel tablets in healthy male volunteers

4区 · 医学

Article

作者: El Ahmady, O. ; Bustami, R. T. ; Ibrahim, M. ; Hussein, A. M.

A randomized, two-way, crossover, bioequivalence study in 32 fasting, healthy, male volunteers was conducted to compare two brands of clopidogrel 75 mg tablets, Thrombo (EIPICO, Egypt) as test and Plavix (Sanofi Pharma/Bristol-Myers Squibb, Paris, France) as reference. The study was performed in a Pharmaceutical Research Unit (PRU) using HPLC/ MS-MS. Arithmetic means for clopidogrel test versus reference formulation, respectively were for Cmax (4.39 +/- 2.58 vs. 4.30 +/- 2.65) ng/ml, AUC0-t (11.98 +/- 9.82 vs. 12.01 +/- 9.46) ng.h/ml, AUC0- yen (12.43 +/- 9.94 vs. 12.49 +/- 9.58) ng.h/ml, t1/2 (6.06 +/- 3.87 vs. 5.87 +/- 2.47) h and the medians for tmax (1 h vs. 0.75 h). Arithmetic means for clopidogrel carboxylic acid metabolite were Cmax (3.75 +/- 1.19 vs. 3.51 +/- 0.97) microg/ml AUC0-t (9.18 +/- 2.36 vs. 9.17 +/- 2.06) microg.h/ml, AUC0- yen (9.72 +/- 2.4 vs. 9.80 +/- 2.21) microg.h/ml , and t1/2 (6.43 +/- 3.52 vs. 6.33 +/- 1.71) h for test versus reference formulation respectively and there was no difference in the medians for tmax (0.75 h). The parametric 90% confidence intervals for the mean of the difference between log-transformed values were within the accepted range for bioequivalence of 80 - 125% as proposed by the US-FDA , namely for clopidogrel (90.66% - 109.66%), (90.63% - 109.73%), and (93.19% - 115.37%) for AUC0-t, AUC0- yen, and Cmax, respectively and also for clopidogrel carboxylic acid metabolite (94.90 - 104.19) , (94.04 - 103.86) and (96.47 - 114.79) for AUC0-t , AUC0- yen, and Cmax, respectively. Thus there was no significant difference between these values and therefore the two products can be considered bioequivalent.

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