Spray-Dried Formulation of Epicertin, a Recombinant Cholera Toxin B Subunit Variant That Induces Mucosal Healing
Epicertin (EPT) is a recombinant variant of the cholera toxin B subunit, modified with a C-terminal KDEL endoplasmic reticulum retention motif. EPT has therapeutic potential for ulcerative colitis treatment. Previously, orally administered EPT demonstrated colon epithelial repair activity in dextran...
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doaj-9059f402df6d4d4aad9515032ad0743a2021-04-18T23:01:36ZengMDPI AGPharmaceutics1999-49232021-04-011357657610.3390/pharmaceutics13040576Spray-Dried Formulation of Epicertin, a Recombinant Cholera Toxin B Subunit Variant That Induces Mucosal HealingMicaela A. Reeves0Joshua M. Royal1David A. Morris2Jessica M. Jurkiewicz3Nobuyuki Matoba4Krystal T. Hamorsky5Department of Pharmacology and Toxicology, University of Louisville School of Medicine, Louisville, KY 40202, USADepartment of Pharmacology and Toxicology, University of Louisville School of Medicine, Louisville, KY 40202, USAJames Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY 40202, USAJames Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY 40202, USADepartment of Pharmacology and Toxicology, University of Louisville School of Medicine, Louisville, KY 40202, USAJames Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY 40202, USAEpicertin (EPT) is a recombinant variant of the cholera toxin B subunit, modified with a C-terminal KDEL endoplasmic reticulum retention motif. EPT has therapeutic potential for ulcerative colitis treatment. Previously, orally administered EPT demonstrated colon epithelial repair activity in dextran sodium sulfate (DSS)-induced acute and chronic colitis in mice. However, the oral dosing requires cumbersome pretreatment with sodium bicarbonate to conserve the acid-labile drug substance while transit through the stomach, hampering its facile application in chronic disease treatment. Here, we developed a solid oral formulation of EPT that circumvents degradation in gastric acid. EPT was spray-dried and packed into enteric-coated capsules to allow for pH-dependent release in the colon. A GM1-capture KDEL-detection ELISA and size-exclusion HPLC indicated that EPT powder maintains activity and structural stability for up to 9 months. Capsule disintegration tests showed that EPT remained encapsulated at pH 1 but was released over 180 min at pH 6.8, the approximate pH of the proximal colon. An acute DSS colitis study confirmed the therapeutic efficacy of encapsulated EPT in C57BL/6 mice upon oral administration without gastric acid neutralization pretreatment compared to vehicle-treated mice (<i>p</i> < 0.05). These results provide a foundation for an enteric-coated oral formulation of spray-dried EPT.https://www.mdpi.com/1999-4923/13/4/576cholera toxin B subunitepicertinspray-dryingpharmaceutical formulationbiopharmaceuticalsulcerative colitis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Micaela A. Reeves Joshua M. Royal David A. Morris Jessica M. Jurkiewicz Nobuyuki Matoba Krystal T. Hamorsky |
spellingShingle |
Micaela A. Reeves Joshua M. Royal David A. Morris Jessica M. Jurkiewicz Nobuyuki Matoba Krystal T. Hamorsky Spray-Dried Formulation of Epicertin, a Recombinant Cholera Toxin B Subunit Variant That Induces Mucosal Healing Pharmaceutics cholera toxin B subunit epicertin spray-drying pharmaceutical formulation biopharmaceuticals ulcerative colitis |
author_facet |
Micaela A. Reeves Joshua M. Royal David A. Morris Jessica M. Jurkiewicz Nobuyuki Matoba Krystal T. Hamorsky |
author_sort |
Micaela A. Reeves |
title |
Spray-Dried Formulation of Epicertin, a Recombinant Cholera Toxin B Subunit Variant That Induces Mucosal Healing |
title_short |
Spray-Dried Formulation of Epicertin, a Recombinant Cholera Toxin B Subunit Variant That Induces Mucosal Healing |
title_full |
Spray-Dried Formulation of Epicertin, a Recombinant Cholera Toxin B Subunit Variant That Induces Mucosal Healing |
title_fullStr |
Spray-Dried Formulation of Epicertin, a Recombinant Cholera Toxin B Subunit Variant That Induces Mucosal Healing |
title_full_unstemmed |
Spray-Dried Formulation of Epicertin, a Recombinant Cholera Toxin B Subunit Variant That Induces Mucosal Healing |
title_sort |
spray-dried formulation of epicertin, a recombinant cholera toxin b subunit variant that induces mucosal healing |
publisher |
MDPI AG |
series |
Pharmaceutics |
issn |
1999-4923 |
publishDate |
2021-04-01 |
description |
Epicertin (EPT) is a recombinant variant of the cholera toxin B subunit, modified with a C-terminal KDEL endoplasmic reticulum retention motif. EPT has therapeutic potential for ulcerative colitis treatment. Previously, orally administered EPT demonstrated colon epithelial repair activity in dextran sodium sulfate (DSS)-induced acute and chronic colitis in mice. However, the oral dosing requires cumbersome pretreatment with sodium bicarbonate to conserve the acid-labile drug substance while transit through the stomach, hampering its facile application in chronic disease treatment. Here, we developed a solid oral formulation of EPT that circumvents degradation in gastric acid. EPT was spray-dried and packed into enteric-coated capsules to allow for pH-dependent release in the colon. A GM1-capture KDEL-detection ELISA and size-exclusion HPLC indicated that EPT powder maintains activity and structural stability for up to 9 months. Capsule disintegration tests showed that EPT remained encapsulated at pH 1 but was released over 180 min at pH 6.8, the approximate pH of the proximal colon. An acute DSS colitis study confirmed the therapeutic efficacy of encapsulated EPT in C57BL/6 mice upon oral administration without gastric acid neutralization pretreatment compared to vehicle-treated mice (<i>p</i> < 0.05). These results provide a foundation for an enteric-coated oral formulation of spray-dried EPT. |
topic |
cholera toxin B subunit epicertin spray-drying pharmaceutical formulation biopharmaceuticals ulcerative colitis |
url |
https://www.mdpi.com/1999-4923/13/4/576 |
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