Human lipoxygenases (LOXs) are enzymes involved in catalyzing the oxidation of polyunsaturated fatty acids to provide the corresponding bioactive hydroxyeicosatetraenoic acid (HETE) metabolites as the end product. These eicosanoid signaling molecules are involved in a number of physiologic responses such as platelet aggregation, inflammation and cell proliferation. ML355 is a potent and selective inhibitor of human 12-lipoxygenase with nM potency. ML355 has been shown to decrease calcium mobilization and PAR-4 induced platelet aggregation in patient derived human platelets and to significantly inhibit AA/IONO-induced (Arachidonic acid and calcium ionophore) 12-HETE (a downstream target of the 12-LOX protein) in mouse BTC3 cells and human islets at 10 µM. Moreover, ML355 demonstrated excellent microsomal stability with both rat (T1/2 >30 minutes) and mouse (T1/2 >300 minutes) and was found to be stable to mouse plasma over a 2 hour period (100% remaining). In vivo PK studies where ML355 was administered as a solution via IV (3 mpk) and PO (30 mpk) demonstrated that ML355 is orally bioavailable (%F = 20) with good half-life (T1/2 = 2.9 hours)[1].
ML355 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Human platelets
25 µM
ML355 treatment significantly decreased 12-HETE formation in thrombin-stimulated human platelets.
ML355 (25 µM) more potently inhibited thrombin-mediated platelet aggregation compared to aspirin (ASA, 100 µM). The combination of ML355 and ASA did not result in an additive inhibitory effect.
ML355 (100 µM) strongly inhibited platelet aggregation induced by PAR1-AP, PAR4-AP, and collagen, and the inhibitory effect diminished with decreasing concentration of ML355.
ML355 dose-dependently inhibited human platelet aggregation induced by thrombin. At low thrombin concentration (0.25 nM), ML355 completely blocked platelet aggregation; at 0.5 nM thrombin, ML355 inhibited 50-60% of platelet aggregation; at 1 nM thrombin, ML355 inhibited about 25% of platelet aggregation. High concentrations of thrombin (>2.5 nM) could reverse the inhibitory effect of ML355.
Simulated the fatty liver environment with IRI in vitro, showing a significant increase in 12-HETE levels.
J Exp Clin Cancer Res. 2019 Dec 12;38(1):489.
mouse platelets
50 µg/ml
5, 15, 30, 60 min
To study the uptake of sHDL by mouse platelets, results showed that sHDL was internalized by platelets within 5 min and reached saturation after 15 min.
Sci Adv. 2020 Dec 4;6(49):eabd0130.
human platelets
10 µM
15 min
To investigate the inhibitory effect of ML355-sHDL on human platelet aggregation, results showed that ML355-sHDL exhibited the strongest inhibition of platelet aggregation.
Sci Adv. 2020 Dec 4;6(49):eabd0130.
HK-2 cells
100 μg/mL
24 h
To evaluate the inhibitory effect of ML355 on GEN-induced oxidative stress, results showed that ML355 significantly reduced ROS generation.
JCI Insight. 2022 Mar 22;7(6):e155487.
ML355 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
DBA2 mice
LUSC xenograft model
3 mg/kg
Once daily for 24 days
ML355 significantly inhibited L1-FGGY-driven tumor growth and reversed the immunosuppressive microenvironment.
Mol Cancer. 2022 Jul 16;21(1):147
Mice
FeCl3-induced mesenteric arteriole thrombosis model and laser-induced cremaster arteriole thrombosis model
Oral
15 mg/kg, 30 mg/kg
Twice daily for two days
ML355 treatment significantly impaired thrombus growth and vessel occlusion. In the FeCl3-induced mesenteric arteriole injury model, platelet adhesion, aggregation, and thrombus formation were impaired in ML355-treated mice, and the thrombi were unstable and frequently embolized. In the laser-induced cremaster arteriole thrombosis model, ML355 (3.5 mg/kg and higher doses) significantly inhibited thrombus formation, and the thrombi were smaller and unstable. ML355 had minimal impact on hemostasis, with no significant increase in hemostatic plug formation or bleeding.
Single dose, continued until the end of the experiment
ML355 reduced HCC recurrence in fatty liver by inhibiting the ALOX12-12-HETE pathway.
J Exp Clin Cancer Res. 2019 Dec 12;38(1):489.
Mice
Laser-induced cremaster arteriole thrombosis model
Intravenous injection
1.5 mg/kg
Single dose, lasting 24 hours
To investigate the inhibitory effect of ML355-sHDL on thrombus formation in mice, results showed that ML355-sHDL significantly inhibited thrombus formation without impairing normal hemostasis.
Sci Adv. 2020 Dec 4;6(49):eabd0130.
Mice
MIOX-Tg mice
Intraperitoneal injection
3 mg/kg
Once daily for 7 days
To evaluate the protective effect of ML355 on GEN-induced kidney injury, results showed that ML355 significantly reduced tubular injury and inflammatory response.
JCI Insight. 2022 Mar 22;7(6):e155487.
Spontaneously hypertensive rats (SHR)
Renal ischemia-reperfusion injury model
Intraperitoneal injection
30 mg/kg
Every other day up to 7 days
ML355 treatment lowered levels of 12-HETE, resulting in reduced renal lipid peroxidation, ER stress, tubular cell death, and inflammation, and improved renal function recovery.
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