生物活性

MG-101 is an inhibitor of Calpain 1 (Ki = 190 nM), Calpain 2 (Ki = 220 nM), cathepsin B (Ki = 150 nM), and cathepsin L (Ki = 500 pM). It inhibits neutral cysteine proteases and proteasome (Ki = 6 μM). It inhibits apoptosis in thymocytes and metamyelocytes. MG-101 also inhibits cell cycle progression at the G1/S border and metaphase/anaphase in CHO cells by inhibiting cyclin B degradation. MG-101 inhibits the proteolysis of IκBα and IκBβ by the ubiquitin-proteasome complex. Furthermore, it protects against neuronal damage caused by hypoxia and ischemia. MG-101 also prevents nitric oxide production by activated macrophages by interfering with transcription of the inducible nitric oxide synthase gene.

ALLN inhibits HeLa cell growth with IC50 of 25.10 ± 0.16μM.[1]

Protease-inhibitory activities[2]

体外研究

体内研究

激酶实验

Calpain Assay[3]

Ca2+-dependent protease activity was determined with resorufin-labeled casein as a substrate. Each incubation mixture (final volume of 100μl) contained sample fraction, 0.02% (w/v) casein, 100mM Tris-HCl, pH 7.5, 5mMβ-mercaptoethanol, and either 5mM EGTA, 4mM CaCl2, or 0.7mM CaCl2. After 1 h incubation at 37oC, the reaction was terminated by adding 125μl of 5% (w/v) trichloroacetic acid. Acid-soluble products were mixed with 2 ml of 0.5 M Tris. HC1, pH 8.8, and the fluorescence emission was determined at 584 nm (excitation at 574 nm) (ϵ= 66 mM-1cm-1). Reactions carried out in the presence of 5mM EGTA were taken as background. Protein concentrations were determined by the method of Bradford. Where indicated, 13μM ALLN was added to the incubation mixture.

细胞实验

Primary Retinal Neuronal Culture and ALLN Treatments[4]

Retinas of newborn Sprague–Dawley rats (1-day old) were removed after anesthesia and incubated at 37oC for 20 min in a papain solution (2 mg/ml). Retinal neurons were mechanically dissociated by using a fire-polished Pasteur pipette. The cell suspension was plated at a density of 1.2x 105cells per cm2onto poly-D-lysine (0.01 mg/ ml)-coated flasks or multi-well plates and cultured in a neurobasal medium, supplemented with 2 % B27, in a humidified 5 % CO2 incubator at 37oC and culture media were changed every 2 days. In the CNS, under these culture conditions, more than 90 % of cells in the cultures were neuronal cells. MAP-2 and NeuN were used to label retinal neurons. Retinal ganglion cells (RGCs) were identified by using specific cell markers, Thy-1.1 and Brn-3a. Experiments were performed on the 7th day of neurons in culture. ALLN was dissolved in dimethyl sulfoxide (DMSO) for storage in 10 mM and then further diluted in culture medium to achieve DMSO concentrations below 0.1 %. Primary retinal neurons were treated with ALLN (1, 2.5, 5, 7.5μM) or DMSO (0.1 %) for 6, 12, and 24 h.

MTT Assay

Cell viability was determined by the 3-(4,5-dimethylthiazole-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Following treatment, cells in the 96-well plates were incubated with 10μl MTT (5 mg/ml) at 37oC for 4 h. The supernatant was removed and replaced by 100μl of dimethyl sulfoxide (DMSO), and the cell viability was measured on a microplate reader at 570 nm. Experiments were repeated independently three times in triplicate and data were represented as MTT reductions relative to control.

动物实验

Xenografts[5]

Four-week-old female athymic nude mice were obtained from HFK Bio. When the mice were 6 weeks old, UbcH10-/- cells and parental cells were collected and washed twice with PBS.A total of 5X106cells were resuspended in 0.2mL PBS and inoculated into the flanks of 10 mice. On the 8th day following inoculation, the mice were divided into two groups: the control group was intraperitoneally injected with vehicle (saline and ethanol), and the treatment group was intraperitoneally injected with 200mL ethanol containing 10mg/kg (26μM) ALLN. The tumors were measured every 3 days, and the tumor volumes were calculated using the formula ([length X width2] X 0.5). The tumors were removed and weighed 23 days after injection.

不同实验动物依据体表面积的等效剂量转换表（数据来源于FDA指南）

例如，已知某工具药用于小鼠的剂量为88 mg/kg , 则用于大鼠的剂量换算方法：将88 mg/kg 乘以小鼠的Km系数（3），再除以大鼠的Km系数（6），得到该药物用于大鼠的等效剂量44 mg/kg。

参考文献

[1] Choi HJ, Cui M, Li DY, Song HO, Kim HS, Park H. Anti-malarial activity of new N-acetyl-L-leucyl-L-leucyl-L-norleucinal (ALLN) derivatives against Plasmodium falciparum. Bioorg Med Chem Lett. 2013;23(5):1293-1296.
[2] Hiwasa T ST, Sakiyama S. Cysteine proteinase inhibitors and ras gene products share the same biological activities including transforming activity toward NIH3T3 mouse fibroblasts and the differentiation-inducing activity toward PC12 rat pheochromocytoma cells. Carcinogenesis. . 1990;11(1):75-80.
[more]

体内溶解度

Clinical Trial Information ( data from http://clinicaltrials.gov )

注：以上所有数据均来自公开文献，并不保证对所有实验均有效，数据仅供参考。

相关化合物库

使用AMQUAR产品发表文献后请联系我们