Antitumor efficacy of paclitaxel-loaded polylactide/ poly(ethylene glycol) nanoparticles combination with exercise in tumor-bearing mice.

The nanoparticles (NPs) provide a promising prospect for tumor therapy, and exercise is also becoming readily and accepted as a beneficial adjunct therapy to maintain or enhance quality of life in cancer patients. We investigate the antitumor efficacy of paclitaxel (PT) loaded polylactide/poly(ethylene glycol) NPs (PT-PLA/PEG NPs) under the exercise conditions. Results showed that within the first 7 days, the PT concentration in tumor maintained at a higher level in the PT-PLA/PEG NPs + exercise (PT-PLA/PEG NPs + EX) group as compared with the PT-PLA/PEG NPs group. All the phagocytosis rates of macrophages were significantly decreased below the CON in exercise group. The most significant antitumor effect was observed in PT-PLA/PEG NPs + EX group, demonstrating that the PT-PLA/PEG NPs improved the concentration of PT, and exercise could further increased its therapeutic efficiency for tumor. These researches may provide an effective means for tumor therapy.

A Fit-for-Purpose Method for the Detection of Human Antibodies to Surface-Exposed Components of BMS-986263, a Lipid Nanoparticle-Based Drug Product Containing a siRNA Drug Substance.

ND-L02-s0201/BMS-986263 is a lipid nanoparticle (LNP) drug product containing a heat shock protein 47 (HSP47)-specific small interfering ribonucleic acid (siRNA) and being developed for the treatment of liver and idiopathic pulmonary fibrosis. To address immunogenicity-related issues, we developed a robust, fit-for-purpose (FFP) three-tier electrochemiluminescent (ECL) anti-drug antibody (ADA) assay for the detection of antibodies (Abs) generated to surface-exposed components of BMS-986263. The drug was coated directly on plates, and several Abs specific for polyethylene glycol (PEG) and other surface components were tested for use as positive quality controls (QCs). Following selection of a rabbit monoclonal anti-PEG Ab, the assay was optimized, and various method development challenges specific to the modality and pseudo surrogate rabbit control were addressed. Screening, confirmatory, and titer cut points were validated following a statistical evaluation of 41 individual K2EDTA human plasma samples at a minimum required dilution (MRD) of 100. Assay precision, sensitivity, selectivity, drug tolerance, and hook effect were determined for the rabbit Ab prepared in human K2EDTA plasma matrix. The assay was used to interrogate anti-drug Ab (ADA) responses in normal human subjects who were administered 90 mg of the drug intravenously (IV) once every week for 3 weeks in phase I clinical trials. All pre- and post-dose samples were found to be negative for ADA. Based on these results, we concluded that BMS-986263 is not immunogenic. To the best of our knowledge, this work represents the first ADA method developed and reported for an LNP-based drug product.

Stabilization of the maleate salt of a basic drug by adjustment of microenvironmental pH in solid dosage form.

Tablet formulations of the maleate salt of a basic drug (I) showed a major loss in potency and a lack of mass balance upon storage under accelerated stability testing conditions. No such stability issues were observed in capsules that were compositionally similar, and even the tablet was stable when it was encapsulated in capsule shell. It was identified that the salt converts to its free base form in the microenvironment of the tablet formulation. Studies using radiolabeled drug substance showed that the free base formed in the tablet volatilized under test conditions used and was absorbed in the wall of plastic container. No mass loss was observed with encapsulated tablets since the capsule shell either protected the drug substance from volatilization or trapped any drug substance that volatilized. The conversion of the salt to free base could be related to the pH-solubility profile of the compound where the pH(max) (pH of maximum solubility) was 3.3-3.6, above which the salt would convert to base while no such conversion would occur below this pH. The microenvironmental pH of the tablet was found to be 4.3, favoring the salt-to-base conversion. A stable tablet formulation with shelf-life >3 years was successfully developed by lowering the microenvironemental pH of tablet from 4.3 to <3.0 by adding citric acid to the formulation.

Bioanalysis of dried saliva spot (DSS) samples using detergent-assisted sample extraction with UHPLC-MS/MS detection.

Dried saliva spot (DSS) sampling is a non-invasive sample collection technique for bioanalysis that can be potentially implemented at the patient's home. A UHPLC-MS/MS assay was developed using detergent-assisted sample extraction to quantify BMS-927711, a drug candidate in development for the treatment of migraines, in human DSS. By implementing DSS sampling at the patients' home, the bioanalytical sample collection for pharmacokinetic evaluation can be done at the time of the acute migraine attack without the need for clinical visits. DSS samples were prepared by spotting 15 µL of liquid saliva onto regular Whatman FTA™ DMPK-C cards and verified with a UV lamp (at λ 254 nm or 365 nm) during DSS punching. The 4-mm DSS punches in a 96-well plate were sonicated with 200 µL of [(13)C2, D4]-BMS-927711 internal standard (IS) solution in 20/80 MeOH/water for 10 min, followed by sonication with 50 µL of 100 mM NH4OAc with 1.0% Triton-X-100 (as detergent) prior to liquid-liquid extraction with 600 µL EtOAc/Hexane (90:10). UHPLC-MS/MS was performed with an Aquity(®) UPLC BEH C18 Column (2.1 × 50 mm, 1.7 µm) on a Triple Quad™ 5500 mass spectrometer. The assay was linear with a concentration range from 2.00 to 1000 ng mL(-1) for BMS-927711 in human saliva. The intra- and inter-assay precision was within 8.8% CV, and the accuracy was within ±6.7% Dev of the nominal concentration values. This UHPLC-MS/MS assay has been successfully applied to determine the drug's pharmacokinetics within a clinical study. For the first time, we observed BMS-927711 exposure in human DSS, confirming the suitability of this sampling technique for migraine patients to use at home. Detergent-assisted extraction with Triton-X-100 could be very useful in DSS or other dried matrix spot (DMS) assays to overcome low or inconsistent analyte recovery issues.

Analysis of cationic liposomes by reversed-phase HPLC with evaporative light-scattering detection.

Cationic lipid-mediated drug delivery of small pharmaceutical molecules and biological molecules, such as proteins and DNA, has gained increasing popularity for many in vitro and in vivo applications. For this purpose, 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) is one of the most widely used and efficient cationic lipids. In this work, a simple and rapid reversed-phase HPLC method was developed for the simultaneous determination of cationic lipid DOTAP and neutral co-lipids cholesterol and phosphatidylcholine (DPPC or DSPC) as well as their degradation products in liposome-based drug formulations. Due to the poor UV absorbance of the lipids and their degradation products, an evaporative light-scattering detector (ELSD) was used to monitor the separation. The HPLC separation was achieved using a Phenomenex Luna C18 column at 50 degrees C by a linear gradient elution with methanol-water mobile phase at a flow rate of 2.0mL/min. 0.1% (v/v) trifluoroacetic acid (TFA) was added into the mobile phase to enhance the retaining of the cationic lipid DOTAP. This newly developed method enabled direct analysis of liposomes without solvent lipid extraction, and was validated to be linear, precise, accurate, specific and sensitive. The limit of detection (LOD) and limit of quantitation (LOQ) were determined to be 0.15 and 0.30microg, respectively, for all the four lipids. The method has been successfully employed in a wide range of lipid-based formulation screening, process development and stability testing. Studies of liposome samples under accelerated thermal conditions revealed that the hydrolysis of DOTAP, DPPC and DSPC followed pseudo-first-order kinetics.