BSEP ( IC50 = 4.8 μM ); MRP4 ( IC50 = 6.2 μM ); MDR3 ( IC50 = 7.5 μM ); LPA1

In the lungs of healthy mice, bleomycin-treated mice, and IPF mice, the percent displacement at 0.1 nM is 18%, 24%, and 31%, respectively. The percentages of displacement at 10 nM are 73%, 76%, and 64%, in that order. As a translational research tool, [¹⁸F]BMT-083133, a radioligand that targets LPA1, is designed to measure lung LPA1 engagement of BMS-986020 through in vitro autoradiography (ARG)[4].

Stroke is a leading cause of death. Stroke survivors often suffer from long-term functional disability. This study demonstrated neuroprotective effects of BMS-986020 (BMS), a selective lysophosphatidic acid receptor 1 (LPA1) antagonist under clinical trials for lung fibrosis and psoriasis, against both acute and sub-acute injuries after ischemic stroke by employing a mouse model with transient middle cerebral artery occlusion (tMCAO). BMS-986020 administration immediately after reperfusion significantly attenuated acute brain injuries including brain infarction, neurological deficits, and cell apoptosis at day 1 after tMCAO. Neuroprotective effects of BMS-986020 were preserved even when administered at 3 h after reperfusion. Neuroprotection by BMS against acute injuries was associated with attenuation of microglial activation and lipid peroxidation in post-ischemic brains. Notably, repeated BMS administration daily for 14 days after tMCAO exerted long-term neuroprotection in tMCAO-challenged mice, as evidenced by significantly attenuated neurological deficits and improved survival rate. It also attenuated brain tissue loss and cell apoptosis in post-ischemic brains. Mechanistically, it significantly enhanced neurogenesis and angiogenesis in injured brains. A single administration of BMS provided similar long-term neuroprotection except survival rate. Collectively, BMS provided neuroprotection against both acute and sub-acute injuries of ischemic stroke, indicating that BMS might be an appealing therapeutic agent to treat ischemic stroke.[5]

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BMS-986020 altered bile homeostasis in vivo, yielding elevated systemic bile acids in rats and humans. In contrast, a structurally distinct LPA1 antagonist BMS-986020, at projected clinically relevant concentrations, did not inhibit BSEP (IC50=19.6 µM), MRP4 (>50 µM), or MDR3 (>50 µM) in vitro, or inhibit bile acid efflux in human hepatocytes (≤50 µM). Additionally, BMS-986020 did not increase bile acids in rats or monkeys. In conclusion, the hepatobiliary effects observed with BMS-986020 are likely off-target effects specific to this molecule and not mediated via antagonism of LPA1. These results suggest that structural variations in LPA1 antagonists may result in different safety profiles in patients with IPF and other fibrotic diseases.[2]

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Of 143 randomized patients, 108 completed the 26-week dosing phase. Thirty-five patients discontinued prematurely. Patient baseline characteristics were similar between treatment groups (placebo: n = 47; 600 mg qd: n = 48; 600 mg bid: n = 48). Patients treated with BMS-986020 bid experienced a significantly slower rate of decline in FVC vs placebo (-0.042 L; 95% CI, -0.106 to -0.022 vs -0.134 L; 95% CI, -0.201 to -0.068, respectively; P = .049). Dose-related elevations in hepatic enzymes were observed in both BMS-986020 treatment groups. The study was terminated early because of three cases of cholecystitis that were determined to be related to BMS-986020 after unblinding. Conclusions: BMS-986020 600 mg bid treatment for 26 weeks vs placebo significantly slowed the rate of FVC decline. Both regimens of BMS-986020 were associated with elevations in hepatic enzymes[2].

TUNEL Assay[5]
To determine effects of BMS-986020 on cell apoptosis, TUNEL immunoassay was performed at 1 day and 15 days after tMCAO using an in-situ cell death detection kit according to the manufacturer’s protocol. Cryostat brain sections were post-fixed in 4% PFA for 10 min and permeabilized with 0.1% sodium citrate in 0.1% Triton X-100 for 2 min on ice. Brain sections were then labelled with TUNEL assay kit for 1 h, washed with PBS, and mounted with VECTASHIELD mounting media. Images were taken with a DP72 camera using a fluorescent microscope.

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Immunohistochemistry Against Iba1 or 4-HNE[5]
To determine the effects of BMS-986020 administration on microglial activation and lipid peroxidation, immunohistochemical analysis was performed as described previously. Briefly, cryostat brain sections were oxidized with 1% H2O2 for 15 min and blocked with 1% fetal bovine serum (FBS) in 0.3% Triton X-100. Sections were then labeled with a rabbit primary antibody against Iba1 (1:500) or 4-hydroxynonenal (4-HNE, 1:500) overnight at 4 °C, further labeled with an appropriate biotinylated secondary antibody (1:200), and then incubated with ABC reagent (1:100, Vector Laboratories). Brain sections were exposed to 3,3’-diaminobenzidine substrate to visualize Iba1- or 4-HNE-positive signals, dehydrated in ascending grade of alcohol, cleared in xylene, and mounted with an Entellan media.

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Double Immunofluorescence Followed by 5-Bromo-2′-Deoxyuridine (BrdU) Incorporation[5]
To determine effects of BMS-986020 administration on neurogenesis and angiogenesis, BrdU/DCX- and BrdU/CD31-double immunofluorescence assays were performed as described previously. In brief, BrdU (50 mg/kg in PBS, i.p.) was administered to mice at 13 and 14 days after tMCAO challenge for four times at 12 h interval. For double immunofluorescence, brain sections were incubated with 2N HCl to denature DNA followed by neutralization with 0.1 M borate buffer. Sections were then blocked with 1% FBS in 0.3% Triton X-100 and simultaneously incubated—with either a rat anti-BrdU (1:400) and a goat anti-DCX (1:100) primary antibodies or a mouse anti-BrdU (1:200A) and a rat anti-CD31 (1:300) primary antibodies—overnight at 4 °C to label newly formed neurons or newly formed blood vessels. Sections were then incubated with respective secondary antibodies (1:1000) conjugated with Cy3 or AF488 and mounted with VECTASHIELD mounting media. Images were obtained using a confocal microscope.

Dulbecco's Modified Eagle Medium (DMEM) + GlutaMax supplemented with 0.4% fetal bovine serum, 37.5 mg/mL Ficoll 70, 25 mg/mL Ficoll 400, and 1% ascorbic acid was used to cultivate human lung fibroblasts in 48-well plates. The cells were stimulated in four replicates with either 1 ng/mL of transforming growth factor beta 1 (TGF-β1) or 20 µM LPA with or without BMS-986020 (0.01, 0.05, 0.1, 0.5, 1, or 5 µM) diluted in dimethyl sulfoxide (DMSO) or vehicle (0.05% DMSO). For twelve days, cells were grown at 37 °C in a 95% O2 and 5% CO2 environment. On days four and eight, the culture media were replaced. Until the biomarker measurements, supernatants were kept at −20 °C in storage. On Day 0 (before starting medication treatment) and Day 12, alamarBlue was utilized to measure cellular metabolism. Lactate dehydrogenase (LDH) release was measured on Days 4, 8, and 12.

After MCA occlusion, mice were randomly assigned into a BMS-986020 or a vehicle (1% DMSO in 10% Tween-80)-administered group. To determine whether BMS-986020 could exert neuroprotective effects against acute brain injuries in tMCAO-challenged mice, BMS-986020 was administered via oral gavage at different dosages (0.5, 2, 5, and 10 mg/kg) immediately after reperfusion. For the time window experiment, BMS-986020 was orally administered at 3 h after reperfusion. To determine long-term neuroprotective effects of BMS-986020 against sub-acute brain injuries, BMS-986020 was orally administered once immediately after reperfusion for the single administration group or daily for the repeated administration group (administration for fourteen consecutive days).[5]

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IM136003 was a phase 2, parallel-arm, multicenter, randomized, double-blind, placebo-controlled trial. Adults with IPF (FVC, 45%-90%; diffusing capacity for carbon monoxide, 30%-80%) were randomized to receive placebo or 600 mg BMS-986020 (once daily [qd] or bid) for 26 weeks. The primary end point was rate of change in FVC from baseline to week 26.[3]

[1]. Lysophospholipid receptors in drug discovery. Exp Cell Res. 2015 May 1;333(2):171-7.

[2]. LPA1 antagonists BMS-986020 and BMS-986234 for idiopathic pulmonary fibrosis: Preclinical evaluation of hepatobiliary homeostasis. European Respiratory Journal.

[3]. Randomized, Double-Blind, Placebo-Controlled, Phase 2 Trial of BMS-986020, a Lysophosphatidic Acid Receptor Antagonist for the Treatment of Idiopathic Pulmonary Fibrosis. Chest. 2018 Nov;154(5):1061-1069.

[4]. Autoradiographic evaluation of [18F]BMT-083133, a lysophosphatidic acid receptor 1 (LPA1) radioligand. The jornal of nuclear medicine.

[5]. BMS-986020, a Specific LPA1 Antagonist, Provides Neuroprotection against Ischemic Stroke in Mice. Antioxidants logo Antioxidants (Basel). 2020 Nov 8;9(11):1097.

BMS-986020 is under investigation in clinical trial NCT02017730 (To Evaluate The Relationship Between Plasma Drug Levels And Receptor Binding in Lung Using PET (Positron Emission Tomography) In Healthy Volunteers).

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BMS-986020 is a small molecule drug with a maximum clinical trial phase of II (across all indications) and has 2 investigational indications.

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Idiopathic pulmonary fibrosis (IPF) is a chronic fibrosing lung disease with limited effective treatment options. The LPA1 pathway has been implicated in the etiology and pathogenesis of IPF and is a promising therapeutic target for fibrotic diseases. LPA1 antagonists, including BMS‑986020 and BMS-986234, are being investigated for IPF. Differences in structure and pharmacology of LPA1 antagonists could impact their efficacy and safety profile. In a Phase 2 trial, BMS-986020 compared with placebo significantly slowed lung function decline but, in some patients, showed hepatobiliary effects; the mechanisms underlying these effects were investigated in vitro and in vivo. In vitro, BMS-986020 inhibits bile acid and phospholipid transporters, BSEP (IC50=4.8 µM), MRP4 (6.2 µM), and MDR3 (7.5 µM), which may reduce bile acid and phospholipid efflux, and alter bile composition and flow. [2]

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Lysophospholipids (LPs), including lysophosphatidic acid (LPA), sphingosine 1-phospate (S1P), lysophosphatidylinositol (LPI), and lysophosphatidylserine (LysoPS), are bioactive lipids that transduce signals through their specific cell-surface G protein-coupled receptors, LPA1-6, S1P1-5, LPI1, and LysoPS1-3, respectively. These LPs and their receptors have been implicated in both physiological and pathophysiological processes such as autoimmune diseases, neurodegenerative diseases, fibrosis, pain, cancer, inflammation, metabolic syndrome, bone formation, fertility, organismal development, and other effects on most organ systems. Advances in the LP receptor field have enabled the development of novel small molecules targeting LP receptors for several diseases. Most notably, fingolimod (FTY720, Gilenya, Novartis), an S1P receptor modulator, became the first FDA-approved medicine as an orally bioavailable drug for treating relapsing forms of multiple sclerosis. This success is currently being followed by multiple, mechanistically related compounds targeting S1P receptor subtypes, which are in various stages of clinical development. In addition, an LPA1 antagonist, BMS-986020 (Bristol-Myers Squibb), is in Phase 2 clinical development for treating idiopathic pulmonary fibrosis, as a distinct compound, SAR100842 (Sanofi) for the treatment of systemic sclerosis and related fibrotic diseases. This review summarizes the current state of drug discovery in the LP receptor field.[1[

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diopathic pulmonary fibrosis (IPF) causes irreversible loss of lung function. The lysophosphatidic acid receptor 1 (LPA1) pathway is implicated in IPF etiology. Safety and efficacy of BMS-986020, a high-affinity LPA1 antagonist, was assessed vs placebo in a phase 2 study in patients with IPF.[3]

C29H26N2O5

482.5271

482.184

C, 72.19; H, 5.43; N, 5.81; O, 16.58

1257213-50-5

BMS-986020 sodium; 1380650-53-2

49792850

White to yellow solid powder

1.3±0.1 g/cm3

664.8±55.0 °C at 760 mmHg

355.9±31.5 °C

0.0±2.1 mmHg at 25°C

1.647

4.99

2

6

8

36

764

1

O([H])C(C1(C2C([H])=C([H])C(C3C([H])=C([H])C(C4=C(C(C([H])([H])[H])=NO4)N([H])C(=O)O[C@]([H])(C([H])([H])[H])C4C([H])=C([H])C([H])=C([H])C=4[H])=C([H])C=3[H])=C([H])C=2[H])C([H])([H])C1([H])[H])=O

GQBRZBHEPUQRPL-LJQANCHMSA-N

InChI=1S/C29H26N2O5/c1-18-25(30-28(34)35-19(2)20-6-4-3-5-7-20)26(36-31-18)23-10-8-21(9-11-23)22-12-14-24(15-13-22)29(16-17-29)27(32)33/h3-15,19H,16-17H2,1-2H3,(H,30,34)(H,32,33)/t19-/m1/s1

1-[4-[4-[3-methyl-4-[[(1R)-1-phenylethoxy]carbonylamino]-1,2-oxazol-5-yl]phenyl]phenyl]cyclopropane-1-carboxylic acid

AM152; AM 152; AM-152; AP-3152 free acid; BMS-986020; 1257213-50-5; AP-3152 free acid; 38CTP01B4L; (R)-1-(4'-(3-Methyl-4-(((1-phenylethoxy)carbonyl)amino)isoxazol-5-yl)-[1,1'-biphenyl]-4-yl)cyclopropane-1-carboxylic acid; Cyclopropanecarboxylic acid, 1-[4'-[3-methyl-4-[[[(1R)-1-phenylethoxy]carbonyl]amino]-5-isoxazolyl][1,1'-biphenyl]-4-yl]-; UNII-38CTP01B4L; 1-[4-[4-[3-methyl-4-[[(1R)-1-phenylethoxy]carbonylamino]-1,2-oxazol-5-yl]phenyl]phenyl]cyclopropane-1-carboxylic acid; BMS-986020; BMS986020; BMS 986020

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: 97~125 mg/mL (201.0~259.1 mM)
Ethanol: ~97 mg/mL

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.31 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (4.31 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)