2-Carba-lysophosphatidic acid is a novel \u03b2-lysophosphatidic acid analogue with high potential for lysophosphatidic acid receptor activation and autotaxin inhibition

Keiko Fukasawa,Kimiko Murakami-Murofushi,Takatsugu Hirokawa,Akiharu Uwamizu,Mari Gotoh,Keisuke Yoshikawa,Masaki Ishikawa,Yoshibumi Shimizu,Kensuke Iwasa,Shinji Yamamoto,Junken Aoki

doi:10.1038/s41598-021-96931-2

Abstract

Cyclic phosphatidic acid (cPA) is a naturally occurring phospholipid mediator that, along with its chemically stabilized analogue 2-carba-cyclic phosphatidic acid (2ccPA), induces various biological activities in vitro and in vivo. Although cPA is similar to lysophosphatidic acid (LPA) in structure and synthetic pathway, some of cPA biological functions apparently differ from those reported for LPA. We previously investigated the pharmacokinetic profile of 2ccPA, which was found to be rapidly degraded, especially in acidic conditions, yielding an unidentified compound. Thus, not only cPA but also its degradation compound may contribute to the biological activity of cPA, at least for 2ccPA. In this study, we determined the structure and examined the biological activities of 2-carba-lysophosphatidic acid (2carbaLPA) as a 2ccPA degradation compound, which is a type of β-LPA analogue. Similar to LPA and cPA, 2carbaLPA induced the phosphorylation of the extracellular signal-regulated kinase and showed potent agonism for all known LPA receptors (LPA1–6) in the transforming growth factor-α (TGFα) shedding assay, in particular for LPA3 and LPA4. 2carbaLPA inhibited the lysophospholipase D activity of autotaxin (ATX) in vitro similar to other cPA analogues, such as 2ccPA, 3-carba-cPA, and 3-carba-LPA (α-LPA analogue). Our study shows that 2carbaLPA is a novel β-LPA analogue with high potential for the activation of some LPA receptors and ATX inhibition.

Highlights

Cyclic phosphatidic acid is a naturally occurring phospholipid mediator that, along with its chemically stabilized analogue 2-carba-cyclic phosphatidic acid (2ccPA), induces various biological activities in vitro and in vivo
The degradation compound was purified by extraction from one spot on a thin layer chromatography (TLC) plate and its purity was confirmed by QqTOF along with liquid chromatography coupled to triple quadrupole mass spectrometry (LC-QqQ)
These results suggested that the structure of the degradation compound was open at the position where the oxygen in the phosphate group was present owing to the strong acidic treatment of 2ccPA (Fig. 2a)

Summary

Introduction

Cyclic phosphatidic acid (cPA) is a naturally occurring phospholipid mediator that, along with its chemically stabilized analogue 2-carba-cyclic phosphatidic acid (2ccPA), induces various biological activities in vitro and in vivo. We determined the structure and examined the biological activities of 2-carba-lysophosphatidic acid (2carbaLPA) as a 2ccPA degradation compound, which is a type of β-LPA analogue. As a continuing investigation to examine the biological activities of 2ccPA, we identified the 2ccPA degradation compound using quadrupole time-of-flight mass spectrometry (QqTOF) and examined its biological activities, including its effect on the phosphorylation of the extracellular signal-regulated kinase (ERK), its activity toward six known LPA receptors ( LPA1–6), and its inhibitory activity against lysoPLD activity of ATX along with docking simulation with ATX

Methods

Results

Conclusion