Effect of Arginine on the Aggregation of Protein in Freeze-Dried Formulations Containing Sugars and Polyol: 1—Formulation Development

Abstract

L-arginine was introduced into protein-based freeze-dried formulations to study the ability of arginine to reduce/prevent from protein aggregation during manufacturing, storage and reconstitution of lyophilized protein-based pharmaceuticals. As L-arginine is known to be very hygroscopic, additional excipients which could provide a moisture buffering capacity need to be introduced into the formulation. In the first part of our study-excipient formulation development-the screening of a number of sugars/polyols has been done in order to select the best combination of excipients that, in a complex with L-arginine, can (i) produce freeze-dried cakes with elegant appearance, adequate mechanical properties and reconstitution times, and (ii) resist/minimise the moisture sorption. Various freeze-dried cakes containing L-arginine in combination with mannitol, trehalose, lactose and sucrose were produced and analysed by TGA, DSC, texture analysis, moisture sorption, cake shrinkage, TVIM and SEM. The non-linear dependencies of the physicochemical properties of the freeze-dried cakes on the sugar-to-mannitol ratios were found. The best combinations of excipients (L-arginine, mannitol and trehalose) were selected to be used in the second part of this work, in which the impact of each selected formulation will be studied in relation to the aggregation of a protein.

Effect of Arginine on the Aggregation of Protein in Freeze-Dried Formulations Containing Sugars and Polyol: II. BSA Reconstitution and Aggregation

“…The present paper continues our study (38) on the effect of arginine on protein aggregation during the manufacturing process and storage. The ultimate aim of this study was to assess the ability of arginine to prevent/reduce protein aggregation caused by the various stresses encountered during lyophilisation, storage and/or reconstitution of a lyophilized product.…”

“…It is known that the extent of protein aggregation in the solid state is related to both the residual water, at the end of the process cycle, and to the amount of water absorbed by the protein on storage, and is due to the impact of water on the conformational dynamics of the protein molecule (39,40). Therefore, the first part (38) of our study-excipient selection (formulation development)-was devoted to the rational design and optimization of a formulation that can resist or, at least, significantly reduce the moisture uptake by lyophilized samples containing L-arginine.…”

“…Analysis of a number of quality attributes (i.e. moisture uptake, cake shrinkage, mechanical properties, internal structure) of the lyophilized 3-excipient formulations (containing L-arginine + mannitol + trehalose) showed that the property dependencies on the trehalose content were non-linear (38). Therefore, in the final part of our study, reported in this article, several trehalose-to-mannitol ratios were selected for test in the…”

Application of Texture Analysis Technique in Formulation Development of Lyophilized Orally Disintegrating Tablets Containing Mannitol, Polyvinylpyrrolidone and Amino Acids

“…It was shown by us previously [39], that mannitol in a combination with amino acids (in particular, Larginine) forms very elegant freeze-dried cakes. The present results also show that addition of small amounts of mannitol (~ 20 wt%) helps improving the external appearance and producing visually elegant lyophilized ODTs comprising PVP and amino acids.…”

“…The presence of mannitol also reduces the moisture uptake by the freeze-dried formulations as mannitol has very good moisture-resistant properties even complexed with highly hygroscopic materials (for instance, L-arginine hydrochloride [39]). The moisture uptake by the ODTs studied was shown to be dependent mostly on the PVP content and increased with the PVP concentration in the formulations (Fig.…”