In Vitro and In Vivo Evaluation of 6-O-α-Maltosyl-β-Cyclodextrin as a Potential Therapeutic Agent Against Niemann-Pick Disease Type C.

Akira Ishii,Shuichi Nakahara,Yuki Kondo,Kazutaka Higaki ,Nushrat Yasmin,Taishi Higashi,Yoshiki Yamada ,Atsushi Ichikawa,Kousaku Ohno,Takeo Tanaka ,Keiichi Motoyama,Muneaki Matsuo,Madoka Fukaura,Yasuyo Okada,Tetsumi Irie,Daisuke Iohara,Yoichi Ishitsuka,J Nishikawa ,Fumitoshi Hirayama,Naomi Nakagata

doi:10.3390/ijms20051152

Abstract

Niemann-Pick disease Type C (NPC) is a rare lysosomal storage disease characterized by the dysfunction of intracellular cholesterol trafficking with progressive neurodegeneration and hepatomegaly. We evaluated the potential of 6-O-α-maltosyl-β-cyclodextrin (G2-β-CD) as a drug candidate against NPC. The physicochemical properties of G2-β-CD as an injectable agent were assessed, and molecular interactions between G2-β-CD and free cholesterol were studied by solubility analysis and two-dimensional proton nuclear magnetic resonance spectroscopy. The efficacy of G2-β-CD against NPC was evaluated using Npc1 deficient Chinese hamster ovary (CHO) cells and Npc1 deficient mice. G2-β-CD in aqueous solution showed relatively low viscosity and surface activity; characteristics suitable for developing injectable formulations. G2-β-CD formed higher-order inclusion complexes with free cholesterol. G2-β-CD attenuated dysfunction of intercellular cholesterol trafficking and lysosome volume in Npc1 deficient CHO cells in a concentration dependent manner. Weekly subcutaneous injections of G2-β-CD (2.9 mmol/kg) ameliorated abnormal cholesterol metabolism, hepatocytomegaly, and elevated serum transaminases in Npc1 deficient mice. In addition, a single cerebroventricular injection of G2-β-CD (21.4 μmol/kg) prevented Purkinje cell loss in the cerebellum, body weight loss, and motor dysfunction in Npc1 deficient mice. In summary, G2-β-CD possesses characteristics favorable for injectable formulations and has therapeutic potential against in vitro and in vivo NPC models.

Highlights

Niemann-Pick disease type C (NPC) is a recessive disorder caused by mutations of the NPC1 (~95% of patients) or NPC2 gene [1]
We previously reported that G2-β-CD was internalized in Npc1 deficient Chinese hamster ovarian (CHO) cells and that it reached the lysosome [16]
This result suggests that the secondary hydroxyl side of G2-β-CD is primarily directed to the dimethyl terminus of free cholesterol (FC) and includes the steroid skeleton in the case of the 1:1 molar complex

Summary

Introduction

Niemann-Pick disease type C (NPC) is a recessive disorder caused by mutations of the NPC1 (~95% of patients) or NPC2 gene [1]. Transmembrane NPC1 and soluble luminal NPC2 proteins interact with unesterified free cholesterol (FC) and have an important role in intracellular cholesterol trafficking from the late endosome/lysosome to other organelles including the endoplasmic reticulum [2,3]. Cellular insufficiency such as cholesterol trafficking dysfunction (FC accumulation and shortage of esterified cholesterol (EC)) increases lysosomal volume, and autophagy malfunction occurs in patients with NPC, causing NPC-related manifestations including hepatosplenomegaly and central nerve dysfunction.

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Results

Conclusion