Pharmaceutical Integrity of Lyophilized Methemoglobin-Albumin Clusters after Reconstitution

Yuto Suzuki; Kazuaki Taguchi; Wataru Okamoto; Yuki Enoki; Teruyuki Komatsu; Kazuaki Matsumoto

doi:10.1021/acsomega.3c01054

Pharmaceutical Integrity of Lyophilized Methemoglobin-Albumin Clusters after Reconstitution

ACS Omega. 2023 Jun 15;8(25):22589-22595. doi: 10.1021/acsomega.3c01054. eCollection 2023 Jun 27.

Authors

Yuto Suzuki¹, Kazuaki Taguchi¹, Wataru Okamoto², Yuki Enoki¹, Teruyuki Komatsu², Kazuaki Matsumoto¹

Affiliations

¹ Faculty of Pharmacy, Keio University, 1-5-30 Shibakoen, Minato-ku, Tokyo 105-8512, Japan.
² Department of Applied Chemistry, Faculty of Science and Engineering, Chuo University, 1-13-27 Kasuga, Bunkyo-ku, Tokyo 112-8551, Japan.

Abstract

Covalent attachment of a ferric hemoglobin (metHb) core to three human serum albumin molecules to form metHb-albumin clusters has previously been used to develop an antidote for hydrogen sulfide poisoning. Lyophilization is one of the most effective approaches to preserve protein pharmaceuticals with minimum contamination and decomposition. However, there is concern that lyophilized proteins may undergo pharmaceutical alteration on reconstitution. This study investigated the pharmaceutical integrity of metHb-albumin clusters on lyophilization and reconstitution with three clinically available reconstitution fluids, (i) sterile water for injection, (ii) 0.9% sodium chloride injection, and (iii) 5% dextrose injection. The metHb-albumin clusters retained their physicochemical properties and structural integrity on lyophilization and reconstitution with sterile water for injection or 0.9% sodium chloride injection, along with comparable hydrogen sulfide scavenging ability compared to non-lyophilized metHb-albumin clusters. The reconstituted protein completely rescued lethal hydrogen sulfide poisoning in mice. On the other hand, lyophilized metHb-albumin clusters reconstituted with 5% dextrose injection showed physicochemical changes and a higher mortality rate in mice subjected to lethal hydrogen sulfide poisoning. In conclusion, lyophilization represents a potent preservation method for metHb-albumin clusters if either sterile water for injection or 0.9% sodium chloride injection is used for reconstitution.