Particle size analyses of polydisperse liposome formulations with a novel multispectral advanced nanoparticle tracking technology.

Liposomes are potent adjuvant constituents for licensed vaccines and vaccine candidates and carriers for drug delivery. Depending on the method of preparation, liposomes vary in size distribution, either forming uniform small size vesicles or a heterogeneous mixture of small to large vesicles. Importantly, differences in liposomal size have been demonstrated to induce differential immune responses. Determination of particle size distribution could therefore be crucial for the efficacy and stability of vaccine formulations. We compared the techniques of dynamic light scattering, laser diffraction, and conventional nanoparticle tracking analysis with a novel multispectral advanced nanoparticle tracking analysis (MANTA) for particle size determination of mono- and polydisperse liposomes. MANTA reported an average 146 nm size of monodisperse liposomes but showed a multimodal distribution of polydisperse liposomes with continuous sizes from 50 to 2000 nm. However, approximately 95% of particles were in the size range of 50-1500 nm and only few particles were identified in the 1500-2000 nm range for the investigated volume. Based on our results, we conclude that MANTA is the most suitable approach and can serve as stand-alone technique for particle size characterization of heterogeneous liposome samples in the 50-2000 nm size range.

Novel 4-substituted-N,N-dimethyltetrahydronaphthalen-2-amines: synthesis, affinity, and in silico docking studies at serotonin 5-HT2-type and histamine H1 G protein-coupled receptors.

Syntheses were undertaken of derivatives of (2S,4R)-(-)-trans-4-phenyl-N,N-dimethyl-1,2,3,4-tetrahydronaphthalen-2-amine (4-phenyl-2-dimethylaminotetralin, PAT), a stereospecific agonist at the serotonin 5-HT2C G protein-coupled receptor (GPCR), with inverse agonist activity at 5-HT2A and 5-HT2B GPCRs. Molecular changes were made at the PAT C(4)-position, while preserving N,N-dimethyl substitution at the 2-position as well as trans-stereochemistry, structural features previously shown to be optimal for 5-HT2 binding. Affinities of analogs were determined at recombinant human 5-HT2 GPCRs in comparison to the phylogenetically closely-related histamine H1 GPCR, and in silico ligand docking studies were conducted at receptor molecular models to help interpret pharmacological results and guide future ligand design. In most cases, C(4)-substituted PAT analogs exhibited the same stereoselectivity ([-]-trans>[+]-trans) as the parent PAT across 5-HT2 and H1 GPCRs, albeit, with variable receptor selectivity. 4-(4'-substituted)-PAT analogs, however, demonstrated reversed stereoselectivity ([2S,4R]-[+]-trans>[2S,4R]-[-]-trans), with absolute configuration confirmed by single X-ray crystallographic data for the 4-(4'-Cl)-PAT analog. Pharmacological affinity results and computational results herein support further PAT drug development studies and provide a basis for predicting and interpreting translational results, including, for (+)-trans-4-(4'-Cl)-PAT and (-)-trans-4-(3'-Br)-PAT that were previously shown to be more potent and efficacious than their corresponding enantiomers in rodent models of psychoses, psychostimulant-induced behaviors, and compulsive feeding ('binge-eating').