Tetrahydrocurcumin (HZIV 81-2)

Cell experiment:

Sup-T1 cells are cultured in RPMI 1640 supplemented with 10% FBS and 1% Penicillin/Streptomycin at 37°C and 5% CO22. 2×105 cells/mL are seeded in each well and Tetrahydrocurcumin, Curcumin and Calebin-A, at 0.1, 0.5, 1.0, 5.0, 10.0, 50.0 and 100.0 μM dissolved in DMSO, are added to their respective wells and incubated for 24, 48 and 72 h. The MTS reagent is added and incubated for 4 h. Absorbance is recorded at 490 nm in Synergy HT multi-well plate reader and Gen5 data analysis software[1].

Animal experiment:

Rats[1]Surgically-modified, exposed jugular vein-catheterized, adult male CD Sprague-Dawley rats (250–300 g) ared used. Each rat is placed in a separate metabolic cage and fasted for 12 h prior to dosing with free access to water. On the day of experiment, the animals (N=3) receive a single dose of Tetrahydrocurcumin by oral gavage (500 mg/kg) in a volume not exceeding 1 mL. Animals have free access to water pre- and post-dosing, and food is provided 2 hours post-dosing. A series of blood samples (0.3 mL) are collected at 0, 15 and 30 min, and 1, 2, 4, 6, 12, 24, 48 and 72 h post-dose. At 72 h after administration, the animals are euthanized and exsanguinated. Immediately after each blood collection time point (except the terminal point), the cannula is flushed with 0.3 mL of 0.9% saline to replenish the collected blood volume. The dead volume of the cannula is replaced with sterile heparin/50% dextrose catheter lock solution to maintain the patency of the cannula as advised in the technical sheet supplied with the animals from Charles River. Following centrifugation of blood samples at 15,000 rpm for 5 min, serum is collected and placed into 2 mL tubes at -20°C until further analysis. Urine samples are collected at 0, 2, 6, 12, 24, 48 and 72 h post-dose and placed in 15 mL tubes. The exact urine volume of each sample is recorded then stored at -20°C until further analysis[1].

References:

[1]. Novaes JT, et al. Disposition, Metabolism and Histone Deacetylase and Acetyltransferase Inhibition Activity of Tetrahydrocurcumin and Other Curcuminoids. Pharmaceutics. 2017 Oct 12;9(4). pii: E45.