Isolation, Physicochemical Characterization, and Biological Properties of Inotodiol, the Potent Pharmaceutical Oxysterol from Chaga Mushroom

Abstract

Inotodiol, an oxysterol found only in Chaga mushroom, has received attention from the pharmaceutical industry due to its strong antioxidant and anti-allergic activities. However, the production of inotodiol is still challenging, and its fundamental properties have yet to be investigated. This study aims to develop an efficient method to produce high-purity inotodiol from Chaga mushroom. Then, pure inotodiol was used to assess its physicochemical properties and biological activities. By optimizing the solvent used for extraction and purification, a new method to produce inotodiol was developed with high purity (>97%) and purification yield (33.6%). Inotodiol exhibited a melting point (192.06 °C) much higher than lanosterol and cholesterol. However, the solubility of inotodiol in organic solvents was notably lower than those of the other two sterols. The difference in the hydroxyl group at C-22 of inotodiol has shown the distinctive physicochemical properties of inotodiol compared with cholesterol and lanosterol. Based on those findings, a nonionic surfactant-based delivery system for inotodiol was developed to improve its bioavailability. The inotodiol microemulsion prepared with 1–2% Tween-80 exhibited homogenous droplets with an acceptable diameter (354 to 217 nm) and encapsulation efficiency (85.6–86.9%). The pharmacokinetic analysis of inotodiol microemulsion in oral administration of 4.5 mg/kg exhibited AUC0–24h = 341.81 (ng·h/mL), and Cmax = 88.05 (ng/mL). Notably, when the dose increased from 4.5 to 8.0 mg/kg, the bioavailability of inotodiol decreased from 41.32% to 33.28%. In a mouse model of sepsis, the serum level of interleukin-6 significantly decreased, and the rectal temperature of mice was recovered in the inotodiol emulsion group, indicating that inotodiol microemulsion is an effective oral delivery method. These results could provide valuable information for applying inotodiol in functional food, cosmetic, and pharmaceutical industries.

Protective Effects of Lanostane Triterpenoids from Chaga Mushroom in Human Keratinocytes, HaCaT Cells, against Inflammatory and Oxidative Stresses

“…The product obtained after freeze-drying was designated as inotodiol concentrate, which had approximately 10% inotodiol. To obtain high-purity inotodiol, inotodiol concentrate was further purified using a recycling HPLC system (MPLC, YMCKOREA, Seongnam, Republic of Korea) equipped with reverse-phase chromatography (ODS-AQ, 50 × 500 mm, YMCKOREA) [ 14 ]. Finally, 95% pure inotodiol was produced and used for the in vitro assays.…”

“…Inotodiol, a lanostane-type triterpenoid from the Chaga mushroom, has been used as a traditional medicine for diabetes, cardiovascular disorders, and tuberculosis [ 11 , 12 ]. Numerous studies have investigated inotodiol’s biological activities, and it has been proven to have antitumor, anti-inflammatory, antiallergic, antiviral, antiaging, and antioxidant properties [ 13 , 14 , 15 , 16 ]. Recently, there has been a movement to identify novel, safe, and effective molecules that can be used for the treatment of various diseases, including inflammatory diseases.…”

Preventive effects of inotodiol on polyinosinic–polycytidylic acid-induced inflammation in human dermal fibroblasts

“…The inotodiol powder was supplied from CARBOEXPERT Inc.,(Daejeon, Republic of Korea) and dissolved in 100 % pure ethanol, and then store at 4 °C before use. Chaga mushroom powder was purchased from Jungwoodang Co., Ltd. (Seoul, Republic of Korea) for Inotodiol extraction and purification, and quality analysis was performed by CarboExpert Inc., according to a previously reported protocol [ 29 ].…”

Chaga Mushroom Extract as a Dual-Action Agent against Microbial and Cancerous Cells: An In Vitro Study

“…Then the extracted solution was separated from chaga powder by centrifugation. The insoluble residue was treated twice again with the same method to increase the yield of the extracted compounds according to Nguyen et al, (2023). Finally, the collected supernatants were evaporated using a vacuum rotary evaporator to obtain the crude extract which was used in further experiments.…”