Enhanced Stability of Vegetal Diamine Oxidase with Trehalose and Sucrose as Cryoprotectants: Mechanistic Insights

Enteric dysfunctions are common for various histamine-related intestinal disorders. Vegetal diamine oxidase (vDAO), an enzyme able to decompose histamine and thus alleviate histamine-related dysfunctions, was formulated in gastro-resistant tablet forms for oral administration as a food supplement and possible therapeutic agent. A major challenge for the use of proteins in the pharmaceutical field is their poor stability. In this study, vDAO was freeze-dried in the absence or in the presence of sucrose or trehalose as cryoprotectants and then formulated as tablets by direct compression. The stability of the obtained preparations was followed during storage at 4 °C and −20 °C for 18 months. In vitro dissolution tests with the vDAO powders formulated as tablets were performed in simulated gastric and in simulated intestinal fluids. The tablets obtained with the powder of the vDAO lyophilized with sucrose or trehalose cryoprotectants offered better protection for enzyme activity. Furthermore, the release of the vDAO lyophilized with the cryoprotectants was around 80% of the total loaded activity (enzyme units) compared to 20% for the control (vDAO powder prepared without cryoprotectants). This report revealed the potential of sucrose and trehalose as cryoprotectants to protect vDAO from freeze-drying stress and during storage, and also to markedly improve the vDAO release performance of tablets obtained with vDAO powders

Enhanced Stability of Vegetal Diamine Oxidase with Trehalose and Sucrose as Cryoprotectants: Mechanistic Insights

Enteric dysfunctions are common for various histamine-related intestinal disorders. Vegetal diamine oxidase (vDAO), an enzyme able to decompose histamine and thus alleviate histamine-related dysfunctions, was formulated in gastro-resistant tablet forms for oral administration as a food supplement and possible therapeutic agent. A major challenge for the use of proteins in the pharmaceutical field is their poor stability. In this study, vDAO was freeze-dried in the absence or in the presence of sucrose or trehalose as cryoprotectants and then formulated as tablets by direct compression. The stability of the obtained preparations was followed during storage at 4 °C and −20 °C for 18 months. In vitro dissolution tests with the vDAO powders formulated as tablets were performed in simulated gastric and in simulated intestinal fluids. The tablets obtained with the powder of the vDAO lyophilized with sucrose or trehalose cryoprotectants offered better protection for enzyme activity. Furthermore, the release of the vDAO lyophilized with the cryoprotectants was around 80% of the total loaded activity (enzyme units) compared to 20% for the control (vDAO powder prepared without cryoprotectants). This report revealed the potential of sucrose and trehalose as cryoprotectants to protect vDAO from freeze-drying stress and during storage, and also to markedly improve the vDAO release performance of tablets obtained with vDAO powders

Synthesis of Metal-Loaded Carboxylated Biopolymers with Antibacterial Activity through Metal Subnanoparticle Incorporation

Carboxymethyl starch (CMS) and carboxymethyl cellulose (CMC) loaded by highly dispersed metal subnanoparticles (MSNPs) showed antibacterial activity against E. coli and B. subtilis strains. Copper and silver were found to act in both cationic and zero-valence forms. The antibacterial activity depends on the metal species content but only up to a certain level. Silver cation (Ag+) showed higher antibacterial activity as compared to Ag0, which was, however, more effective than Cu0, due to weaker retention. The number of carboxyl groups of the biopolymers was found to govern the material dispersion in aqueous media, the metal retention strength and dispersion in the host-matrices. Cation and metal retention in both biopolymers was found to involve interactions with the oxygen atoms of both hydroxyl and carboxyl groups. There exists a ternary interdependence between the Zeta potential (ZP), pH induced by the biocidal agent and its particle size (PS). This interdependence is a key factor in the exchange processes with the surrounding species, including bacteria. Clay mineral incorporation was found to mitigate material dispersion, due to detrimental competitive clay:polymer interaction. This knowledge advancement opens promising prospects for manufacturing metal-loaded materials for biomedical applications