A Novel Highly Potent Autotaxin/ENPP2 Inhibitor Produces Prolonged Decreases in Plasma Lysophosphatidic Acid Formation In Vivo and Regulates Urethral Tension

Abstract

Autotaxin, also known as ectonucleotide pyrophosphatase/phosphodiesterase 2 (ENPP2), is a secreted enzyme that has lysophospholipase D activity, which converts lysophosphatidylcholine to bioactive lysophosphatidic acid. Lysophosphatidic acid activates at least six G-protein coupled recpetors, which promote cell proliferation, survival, migration and muscle contraction. These physiological effects become dysfunctional in the pathology of cancer, fibrosis, and pain. To date, several autotaxin/ENPP2 inhibitors have been reported; however, none were able to completely and continuously inhibit autotaxin/ENPP2 in vivo. In this study, we report the discovery of a highly potent autotaxin/ENPP2 inhibitor, ONO-8430506, which decreased plasma lysophosphatidic acid formation.The IC50 values of ONO-8540506 for lysophospholipase D activity were 6.4–19 nM for recombinant autotaxin/ENPP2 proteins and 4.7–11.6 nM for plasma from various animal species. Plasma lysophosphatidic acid formation during 1-h incubation was almost completely inhibited by the addition of >300 nM of the compound to human plasma. In addition, when administered orally to rats at a dose of 30 mg/kg, the compound demonstrated good pharmacokinetics in rats and persistently inhibited plasma lysophosphatidic acid formation even at 24 h after administration.Smooth muscle contraction is a known to be promoted by lysophosphatidic acid. In this study, we showed that dosing rats with ONO-8430506 decreased intraurethral pressure accompanied by urethral relaxation. These findings demonstrate the potential of this autotaxin/ENPP2 inhibitor for the treatment of various diseases caused by lysophosphatidic acid, including urethral obstructive disease such as benign prostatic hyperplasia.

Regulation of autotaxin expression and secretion by lysophosphatidate and sphingosine 1-phosphate

“…All procedures were performed in accordance with the Canadian Council of Animal Care as approved by the University of Alberta Animal Welfare Committee. ONO-8430506 was prepared at 10 mg/ml in 25 mM sodium hydroxide and heated to 60°C for 5 min to dissolve ( 9,29 ). The solution was passed through a 0.22 m fi lter prior to administration.…”

“…TNF-␣ and interleukin 1 ␤ (IL-1 ␤ ) were from PeproTech (Rocky Hill, NJ). ONO-8430506 was obtained from Ono Pharmaceuticals (Osaka, Japan; patent number WO2012005227) ( 28,29 ) under a materials transfer agreement. S32826 was from Institut de Recherches Servier (Croissy-sur-Seine, France) ( 30 ).…”

“…While these probes may be effi cient for screening potential compounds, positive hits for ATX inhibition or derivative analogs should be verifi ed in the presence of physiological concentrations of LPC, as used in the choline release assay. Physiologically relevant LPC concentrations have been used to validate the in vitro effi cacy of many such inhibitors ( 15,20,21,29,44 ). delipidated serum.…”

Recent advances in targeting the autotaxin-lysophosphatidate-lipid phosphate phosphatase axis in vivo

“…It was the first reported ATX inhibitor to reduce plasma LPA levels in vivo for an extended period [140] . In rat air-pouch models, 30 mg/kg PF-8380 inhibited inflammatory hyperalgesia with the same efficacy as 30 mg/kg naproxen, a routinely used nonsteroidal anti-inflammatory drug Inflammatory hyperalgesia [140] Radiotherapy sensitizer in glioblastoma [146] ONO-8430506 Autotaxin activity inhibitor (competitive) Preclinical IC 50 5 nmol/L 10-30 mg/kg Benign prostatic hyperplasia [147] Reduces breast tumor growth and metastasis and increases sensitization to doxorubicin [76,148] Lpathomab TM LPA monoclonal antibody Preclinical 25 mg/kg Traumatic brain injury [127] AM966 LPA 1 antagonist Preclinical IC 50 17 nmol/L 10 mg/kg Idiopathic pulmonary fibrosis [132] BMS-986020 LPA 1 antagonist Phase II 600 mg/day (patients) Idiopathic pulmonary fibrosis [133] BMS-986202/AM152 LPA 1 antagonist Phase I 20-40 mg/kg Idiopathic pulmonary fibrosis [134] SAR100842 LPA 1/3 antagonist Phase II 20-40 mg/kg Systemic sclerosis [135] Gene overexpression Induced LPP gene expression Preclinical Overexpressed in cancer cells LPP3 overexpression reduces ovarian cancer cell growth [118] LPP1 overexpression reduces tumor growth and metastasis in breast and thyroid cancers [116] (NSAID) [140] . At this concentration, PF-8380 maximally reduced LPA levels in both plasma and at the site of inflammation.…”

“…In this study, inhibition of ATX by PF-8380 abrogated radiationinduced activation of Akt and subsequently decreased tumor vascularity and tumor growth [145] ( Table 1). Another potent ATX inhibitor, ONO-8430506, currently in development by Ono Pharmaceuticals Ltd. (Osaka, Japan), is a tetrahydrocarboline derivative with an IC 50 of 5 nmol/L for plasma ATX activity [147] . Dosing rats orally with 30 mg/kg of this compound potently suppressed ATX plasma activity and LPA concentrations for 24 hours and decreased intraurethral pressure in benign prostatic hyperplasia models [147] ( Table 1).…”