ONO-7300243 (ONO 7300243, ONO7300243) is a novel, potent and selective antagonist of LPA1(Lysophosphatidic Acid Receptor) with an IC50 of 160 nM. ONO-7300243 was identified from a high throughout screening and subsequent structural optimizaiton. ONO-7300243 was found at a high through structural optimization that followed screening. In vivo, ONO-7300243 exhibits good efficacy. LPA is known to elicit different physiological responses via a group of G protein-coupled receptors called LPA1-6. Notably, research revealed that ONO-7300243 exhibited potency comparable to tamsulosin, a α1 adrenoceptor antagonist used clinically to treat dysuria in patients with benign prostatic hyperplasia (BPH). Rats given 30 mg/kg of 17a orally experience a decrease in intraurethral pressure.

Physicochemical Properties

Molecular Formula	C28H31NO5
Molecular Weight	461.55
Exact Mass	461.22
Elemental Analysis	C, 72.86; H, 6.77; N, 3.03; O, 17.33
CAS #	638132-34-0
Related CAS #	638132-34-0
PubChem CID	66775043
Appearance	White to off-white solid powder
Density	1.2±0.1 g/cm3
Boiling Point	688.1±55.0 °C at 760 mmHg
Flash Point	369.9±31.5 °C
Vapour Pressure	0.0±2.3 mmHg at 25°C
Index of Refraction	1.591
LogP	5.59
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	11
Heavy Atom Count	34
Complexity	612
Defined Atom Stereocenter Count	0
SMILES	O=C(C1C([H])=C(C(C([H])([H])[H])=C(C=1[H])OC([H])([H])[H])OC([H])([H])[H])N(C([H])([H])C1C([H])=C([H])C(C([H])([H])C(=O)O[H])=C([H])C=1[H])C([H])([H])C([H])([H])C([H])([H])C1C([H])=C([H])C([H])=C([H])C=1[H]
InChi Key	WGABOZPQOOZAOI-UHFFFAOYSA-N
InChi Code	InChI=1S/C28H31NO5/c1-20-25(33-2)17-24(18-26(20)34-3)28(32)29(15-7-10-21-8-5-4-6-9-21)19-23-13-11-22(12-14-23)16-27(30)31/h4-6,8-9,11-14,17-18H,7,10,15-16,19H2,1-3H3,(H,30,31)
Chemical Name	2-[4-[[(3,5-dimethoxy-4-methylbenzoyl)-(3-phenylpropyl)amino]methyl]phenyl]acetic acid
Synonyms	ONO 7300243; ONO7300243; ONO-7300243
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets	LPA1 ( IC50 = 0.19-0.13 μM )
ln Vitro	In vitro activity: ONO-7300243 demonstrated much stronger effects in vivo (88% inhibition at 10 mg/kg i.d., 62% inhibition at 3 mg/kg i.d.), despite its modest in vitro activity (IC50 = 0.16 μM). ONO-7300243 showed good metabolic stability and membrane permeability against rat liver microsomes[1].
ln Vivo	ONO-7300243 prevented the rise in intraurethral pressure (IUP) caused by LPA up to one hour following dosing in a dose-dependent manner (ID50 = 11.6 mg/kg p.o.). At 10 and 30 mg/kg, significant effects were seen (p<0.05 compared to the vehicle). In conscious rats without LPA stimulation, ONO-7300243 (30 mg/kg, p.o.) significantly reduced the IUP in comparison to the vehicle without changing the mean blood pressure (MBP). ONO-7300243 demonstrated a brief half-life (0.3 h) and a quick clearance (CLtot = 15.9 mL/min/kg at 3 mg/kg i.v.) in a rat pharmacokinetic study[1].
Cell Assay	The 96-well plates were seeded with 2×104 Chinese hamster ovary (CHO) cells that were stably expressing human LPA1. The cells were then cultured in the F-12 Nutrient Mixture (HAM) culture medium, which contained 10% FBS, in a CO2 incubator (37ºC, 5% CO2, 95% air) for two days. Each well was filled with load buffer (culture medium containing 5 µM Fura2-AM, 10 mM HEPES (pH 7.55), and 2.5 mM probenecid), which was then incubated for an hour in a CO2 incubator. The cells were rinsed with room-temperature assay buffer after the load buffer was removed, and then the assay buffer was added. Using a fluorescence drug screening system, the intracellular Ca2+ concentration was tracked in the LPA1 antagonist assay experiment by measuring the ratio of fluorescence intensities (f340/f380) at 500 nm. The cells were treated with lysophosphatidic acid (LPA) at a final concentration of 100 nM following the antagonist pretreatment. After treating the compounds, the peak ratio of LPA was compared to that of the control (DMSO) to determine the antagonists' inhibition rate (%). In addition, the Sigmoid Emax Model was used in a non-linear regression analysis to estimate IC50 values.
Animal Protocol	Formulated in 0.5% methyl cellulose; 3 mg/kg for i.v. or 10 mg/kg for p.o.; i.v. or p.o. Rats
References	[1]. Discovery of ONO-7300243 from a Novel Class of Lysophosphatidic Acid Receptor 1 Antagonists: From Hit to Lead. ACS Med Chem Lett. 2016 Aug 19;7(10):913-918.

Solubility Data

Solubility (In Vitro)

DMSO: 92~100 mg/mL (199.3~216.7 mM) Water: <1 mg/mL Ethanol: ~92 mg/mL (199.3 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 3 mg/mL (6.50 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 30.0 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. Solubility in Formulation 2: ≥ 3 mg/mL (6.50 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 30.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1666 mL	10.8331 mL	21.6661 mL
	5 mM	0.4333 mL	2.1666 mL	4.3332 mL
	10 mM	0.2167 mL	1.0833 mL	2.1666 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.