Tissue distribution and safety evaluation of a claudin-targeting molecule, the C-terminal fragment of Clostridium perfringens enterotoxin.
We previously found that claudin (CL) is a potent target for cancer therapy using a CL-3 and -4-targeting molecule, namely the C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE). Although CL-3 and -4 are expressed in various normal tissues, the safety of this CL-targeting strategy has never been investigated. Here, we evaluated the tissue distribution of C-CPE in mice. Ten minutes after intravenous injection into mice, C-CPE was distributed to the liver and kidney (24.0% and 9.5% of the injected dose, respectively). The hepatic level gradually fell to 3.2% of the injected dose by 3 h post-injection, whereas the renal C-CPE level gradually rose to 46.5% of the injected dose by 6 h post-injection and then decreased. A C-CPE mutant protein lacking the ability to bind CL accumulated in the liver to a much lesser extent (2.0% of the dose at 10 min post-injection) than did C-CPE, but its renal profile was similar to that of C-CPE. To investigate the acute toxicity of CL-targeted toxin, we intravenously administered C-CPE-fused protein synthesis inhibitory factor to mice. The CL-targeted toxin dose-dependently increased the levels of serum biomarkers of liver injury, but not of kidney injury. Histological examination confirmed that injection of CL-targeted toxin injured the liver but not the kidney. These results indicate that potential adverse hepatic effects should be considered in C-CPE-based cancer therapy.
Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan.
Xiangru
X
Li
Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan.
Rie
R
Saeki
Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan.
Akihiro
A
Watari
Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan.
Kiyohito
K
Yagi
Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan. Electronic address: masuo@phs.osaka-u.ac.jp.
Masuo
M
Kondoh
D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology; D002478:Cells, Cultured; D062465:Claudin-3 / Q000378:metabolism*; D062506:Claudin-4 / Q000378:metabolism*; D003016:Clostridium perfringens; D004768:Enterotoxins / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*; D005260:Female; D005347:Fibroblasts; D007413:Intestinal Mucosa / Q000378:metabolism; D007668:Kidney / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008099:Liver / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology*; D011500:Protein Synthesis Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology; D013961:Thyroid Gland / Q000378:metabolism; D014018:Tissue Distribution
D000970:Antineoplastic Agents; D062465:Claudin-3; D062506:Claudin-4; D004768:Enterotoxins; D010446:Peptide Fragments; D011500:Protein Synthesis Inhibitors; D011994:Recombinant Proteins; C035598:enterotoxin, Clostridium
ALT; AST; BSA; BUN; C-CPE; C-terminal fragment of CPE; CL; CPE; Claudin; Clostridium perfringens enterotoxin; FACS; Kidney; Liver; PBS; PSIF; TJ; Tissue distribution; alanine aminotransferase; aspartate aminotransferase; blood urea nitrogen; bovine serum albumin; claudin; fluorescence-activated cell sorter; phosphate-buffered saline; protein synthesis inhibitory factor; tight junction