Nanoparticle-Encapsulated Epirubicin Efficacy in the Inhibition of Growth of Orthotopic Ovarian Patient-Derived Xenograft in Immunocompromised Mice.

Epirubicin hydrochloride (EPI) is an anticancer drug widely used in the treatment of many solid tumors, including ovarian cancer. Because of its anatomical location, ovarian cancer shows symptoms when it is already in an advanced stage and is thus more difficult to treat. Epirubicin hydrochloride kills cancer cells effectively, but its dose escalation is limited by its severe toxicity. By encapsulating epirubicin in dextran-based nanoparticles (POLEPI), we expected to deliver higher and thus clinically more effective doses directly to tumors, where epirubicin would be released and retained longer in the tumor. The antitumor activity of POLEPI compared to EPI was first tested ex vivo in a series of ovarian cancer patient-derived tumor xenografts (PDX). The most promising PDX was then implanted orthotopically into immunocompromised mice, and tumor growth was monitored via magnetic resonance imaging (MRI). Although we succeeded in suppressing the growth of ovarian cancer derived from a patient, in a mouse model by 70% compared to 40% via EPI in 5 days after only one injection, we could not eliminate serious side effects, and the study was terminated prematurely for humane reasons.

In vitro compatibility and stability of admixtures containing etoposide, epirubicin hydrochloride and vindesine sulphate in a single infusion bag.

WHAT IS KNOWN AND OBJECTIVES: The etoposide, doxorubicin hydrochloride, vincristine sulphate, cyclophosphamide and prednisone (EPOCH) chemotherapy regimen is effective in patients with relapsed or refractory non-Hodgkin's lymphoma. However, vincristine and doxorubicin hydrochloride are relatively toxic, leading to neurovirulence and cardiotoxicity, respectively. In this study, we replaced these drugs with vindesine and epirubicin hydrochloride to reduce the cardiotoxicity and evaluated admixtures containing these drugs along with etoposide in a single infusion bag in vitro. METHODS: The appearance and pH of the admixtures were evaluated, and the number of particles was detected. High-performance liquid chromatography was used to measure the concentration and degradation rates of etoposide, epirubicin hydrochloride and vindesine sulphate in each admixture. RESULTS AND DISCUSSION: No precipitation occurred when mixing clinically relevant concentrations of etoposide, epirubicin hydrochloride and vindesine sulphate in a 0.9% NaCl injection solution. Furthermore, the delta pH of the admixtures was ≤0.12 throughout the experiment, and the number of particles (≥10 and ≥25 µm) in the solutions over the 24 hours post-preparation period met USP standards. Etoposide, epirubicin hydrochloride and vindesine sulphate were retained at >96% of their initial concentrations in the admixtures at 25°C over the course of the experiment. Etoposide, epirubicin hydrochloride and vindesine sulphate are compatible when mixed in a 0.9% NaCl injection solution, and the admixtures are stable for at least 24 hours when stored in infusion bags. WHAT IS NEW AND CONCLUSION: This in vitro analysis indicates the suitability of our novel admixtures containing less toxic drug equivalents in a single infusion bag for clinical application.

Synthesis and application of thiol-functionalized magnetic nanoparticles for studying interactions of epirubicin hydrochloride with bovine serum albumin by fluorescence spectrometry.

A novel method was developed for studying the interaction between epirubicin hydrochloride (EPI) and bovine serum albumin (BSA) by fluorescence spectrometry. Fe3 O4 magnetic nanoparticles (MNPs) synthesized and functionalized with thiol group were employed for the immobilization and separation of target BSA in reaction solutions. The concentrations of the non-immobilized BSA and unbound EPI were obtained separately by fluorescence spectrometry. The binding constants (Ka ) and number of binding sites (n) of EPI with BSA were calculated. In this study, the Ka value was 5.05 × 105  L mol-1 , suggesting a strong binding of EPI to BSA, and the n value was 1.15. The effects of common metal ions on Ka of EPI with BSA were also investigated, and the results showed there was clearly bindings between the metal ions and BSA. The precise binding sites of EPI on BSA were determined as being in site I from the competitive displacement experiments. Copyright © 2016 John Wiley & Sons, Ltd.

An Attempt to Shorten Loading Time of Epirubicin into DC Beads® Using Vibration and a Sieve.

PURPOSE: We investigated the possibility of shortening the time required for loading epirubicin into calibrated polyvinyl alcohol-based hydrogel beads (DC Beads®) to be used for transarterial chemoembolization. METHOD: After separating the beads suspended in phosphate-buffered saline (PBS) solution by the use of a sieve (clearance 75 µm), epirubicin hydrochloride (EH) was loaded for 20, 30, or 60 s under vibration into DC beads. The EH loading rate into conventionally prepared (control) beads, i.e., beads loaded for 30 min without vibration, and vibration-loaded beads were calculated from the residual EH concentration in the bead-depleted EH solution. The amount of EH eluted from conventionally and vibration-loaded samples into a PBS solution (pH 7.0) was measured at 15 and 30 min and 1, 2, 6, 12, and 24 h. We also recorded the inhibitory effect of the PBS solution on the loading time. Using frozen sections, the EH load in the beads was evaluated visually under a fluorescence microscope. RESULTS: Spectrophotometry (495 nm) showed that the loading rate was 98.98 ± 0.34, 99.02 ± 0.32, and 99.50 ± 0.11 % with 20-, 30-, and 60-s vibration, respectively. The eluted rate was statistically similar between vibration- and statically loaded (control) beads. The PBS solution hampered EH loading into the beads. Visually, the distribution of EH in conventionally and vibration-loaded DC beads was similar. DISCUSSION: The use of vibration and the removal of PBS solution when epirubicin hydrochloride was loaded into DC beads dramatically shortened the loading time of epirubicin hydrochloride into DC beads.

Surface decorated nanoparticles as surrogate carriers for improved transport and absorption of epirubicin across the gastrointestinal tract: Pharmacokinetic and pharmacodynamic investigations.

Epirubicin (EPI) is a P-gp substrate antracycline analogue which elicits poor oral bioavailability. In the present work, EPI loaded poly-lactide-co-glycolic acid nanoparticles (PLGA-NPs) were prepared by double emulsion approach and superficially decorated with polyethylene glycol (EPI-PNPs) and mannosamine (EPI-MNPs). Average hydrodynamic particle size of EPI-PNPs and EPI-MNPs was found 248.63 ± 12.36 and 254.23 ± 15.16 nm, respectively. Cytotoxicity studies were performed against human breast adenocarcinoma cell lines (MCF-7) confirmed the superiority of EPI-PNPs and EPI-MNPs over free epirubicin solution (EPI-S). Further, confocal laser scanning microscopy (CLSM) and flow cytometric analysis (FACS) demonstrated enhanced drug uptake through EPI-PNPs and EPI-MNPs and elucidated dominance of caveolae mediated endocytosis for NPs uptake. Cellular transport conducted on human colon adenocarcinoma cell line (Caco-2) showed 2.45 and 3.17 folds higher permeability of EPI through EPI-PNPs and EPI-MNPs when compared with EPI-S (p<0.001) while permeability of EPI was found 5.23 and 5.67 folds higher across rat ileum, respectively. Furthermore, pharmacokinetic studies demonstrated 4.7 and 5.57 folds higher oral bioavailability through EPI-PNPs and EPI-MNPs when compared with EPI-S. In addition, both, EPI-PNPs and EMNPs showed tumor suppression comparable to indicated route (i.v. injection). EPI-MNPs showed 1.18 folds higher bioavailability and better tumor suppression than EPI-PNPs.

Improved antitumor activity and reduced cardiotoxicity of epirubicin using hepatocyte-targeted nanoparticles combined with tocotrienols against hepatocellular carcinoma in mice.

Hepatocellular carcinoma (HCC) is the third most common cause of cancer death worldwide. Epirubicin (EPI), an anthracycline derivative, is one of the main line treatments for HCC. However, serious side effects including cardiomyopathy and congestive heart failure limit its long term administration. Our main goal is to develop a delivery strategy that ensures improved efficacy of the chemotherapeutic agent together with reduced cardiotoxicity. In this context, EPI was loaded in chitosan-PLGA nanoparticles linked with asialofetuin (EPI-NPs) selectively targeting hepatocytes. In an attempt to reduce cardiotoxicity, targeted EPI-NPs were coadministered with tocotrienols. EPI-NPs significantly enhanced the antiproliferative effect compared to free EPI as studied on Hep G2 cell line. Nanoencapsulated EPI injected in HCC mouse model revealed higher p53-mediated apoptosis and reduced angiogenesis in the tumor. Combined therapy of EPI-NPs with tocotrienols further enhanced apoptosis and reduced VEGF level in a dose dependent manner. Assessment of cardiotoxicity indicated that EPI-NPs diminished the high level of proinflammatory cytokine tumor necrosis factor-α (TNF-α) as well as oxidative stress-induced cardiotoxicity as manifested by reduced level of lipid peroxidation products (TBARS) and nitric oxide (NO). EPI-NPs additionally restored the diminished level of superoxide dismutase (SOD) and reduced glutathione (GSH) in the heart. Interestingly, tocotrienols provided both antitumor activity and higher protection against oxidative stress and inflammation induced by EPI in the heart. This hepatocyte-targeted biodegradable nanoparticle/tocotrienol combined therapy represents intriguing therapeutic strategy for EPI providing not only superior efficacy but also higher safety levels.

Determination of Drug Loading and Encapsulation Efficiency of Epirubicin Hydrochloride-sorafenib PL-GA Embolic Microspheres by HPLC / 中国药房

OBJECTIVE:To establish a method for the determination of drug loading and encapsulation efficiency of Epirubi-cin hydrochloride-sorafenib-loaded Polylactic Acid-glycolic Acid Polymer(PLGA)embolic microspheres. METHODS:HPLC meth-od was adopted to determine the contents of epirubicin hydrochloride and sorafenib in the preparation,and then drug loading and encapsulation efficiency were calculated by formula. The determination was performed on Phenomenex Luna 5u C8(2) 100A col-umn with mobile phase consisted of methanol-water(containing 0.05% trifluoroacetic acid and 0.14% dium dodecyl sulfate)(75:25,V/V)at the flow rate of 1.0 mL/min. The detection wavelength was set at 252 nm,and the column temperature was maintained at 25℃. The injection volume was 10μL. RESULTS:The linear ranges were 2.020-101.00μg/mL for epirubicin hydrochloride(r=0.9998)and 2.048-102.40 μg/mL for sorafenib(r=0.9997),respectively. The limits of quantification were 3.2970,2.5468 μg/mL, respectively. The detection limits were 0.9891,0.7641 μg/mL,respectively. RSDs of precision,stability and repeatability tests were all less than 2.0%. The recoveries were 96.41%-101.80%(RSD=1.64%,n=9),99.46%-101.45%(RSD=0.70%,n=9),re-spectively. Drug loading of two components in 3 batches of samples were no lower than 1.17%,encapsulation efficiency no lower than 58%. CONCLUSIONS:The method is simple,accurate,can be used to determine drug loading and encapsulation efficiency of Epirubicin hydrochloride-sorafenib PLGA embolic microspheres.

Stability study of epirubicin hydrochloride solutions for injection prepared from three different formulations / 中国药学杂志

OBJECTIVE: To evaluate the stability of epirubicin hydrochloride solutions for clinical use prepared from three different formulations (one concentrated solution and two powders for injection). METHODS: Epirubicin hydrochloride in three formulations was dissolved or diluted in 0.9% sodium chloride to prepare the test solutions with concentration of 0.4mg · mL-1. These solutions were stored at different temperatures and under different shading circumstances, and samples were collected periodically during the storage to evaluate the stability, including visual inspection, pH value and content of epirubicin. RESULTS: All epirubicin solutions were stable when stored at 2-8°C under dark condition for 40 d with average content of greater than 93.76%. When stored atroom temperature with or without light exposure for 40 d, only the solution diluted from the concentrated water solution for injection was stable, as the average content remained above 92.57%. While the solutions prepared from the other two powders for injection were not stable, as the average content fell below 90% at room temperature after 2-7 d. CONCLUSION: Epirubicin solutions prepared from three different formulations are stable when stored either at 2-8°C under dark condition for 40 d or at room temperature for 24 h. The solution prepared from the concentrated water solution for injection is more stable than the solutions prepared from the two powders for injections when stored at room temperature for a long period (40 d).

Preparation and Physicochemical Property of Epirubicin Hydrochloride Solid Lipid Nanoparticles / 中国药房

OBJECTIVE:To prepare epirubicin hydrochloride solid lipid nanoparticle(EPI-SLN) and investigate its physicochemical property. METHODS: EPI-SLN was prepared by ultrasonic dispersion technique with glyceryl behenate as matrix,and the appearance,particle size,? electric potential,and entrapment efficiency of the SLN were evaluated and its stability at 3 month storing at 4 ℃ was investigated. RESULTS: EPI-SLN assumed spherical shape with a particle diameter of (212.8?6.2) nm,? electric potential of (-24.7?0.3) mV and entrapment efficiency of 82 %. The EPI-SLN at 4 ℃ was stable after storing for 3 months,showing no marked change in mean diameter,? electric potential or entrapment efficiency. CONCLUSION: The prepared EPI-SLN is up to the standard.