FPR2 agonist 2 is a potent BBB (blood-brain barrier) permeable (penetrable) FPR2 agonist/activator with EC50 of h-FPR2 of 0.13 µM. FPR2 agonist 2 inhibits proinflammatory cytokine production, counteracts changes in mitochondrial function, and inhibits caspase-3 activity.

Physicochemical Properties

Molecular Formula	C25H20F2N4O2
Molecular Weight	446.448712348938
Exact Mass	446.155
CAS #	2829263-20-7
PubChem CID	163322252
Appearance	White to off-white solid powder
LogP	4
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	5
Heavy Atom Count	33
Complexity	740
Defined Atom Stereocenter Count	1
SMILES	N([C@@H](CC1=CC=C(C#N)C=C1)C(N1C2=C(C=C(F)C=C2)CC1)=O)C(NC1=CC=C(F)C=C1)=O
InChi Key	KCNMTQVKKFREAN-QFIPXVFZSA-N
InChi Code	InChI=1S/C25H20F2N4O2/c26-19-5-8-21(9-6-19)29-25(33)30-22(13-16-1-3-17(15-28)4-2-16)24(32)31-12-11-18-14-20(27)7-10-23(18)31/h1-10,14,22H,11-13H2,(H2,29,30,33)/t22-/m0/s1
Chemical Name	1-[(2S)-3-(4-cyanophenyl)-1-(5-fluoro-2,3-dihydroindol-1-yl)-1-oxopropan-2-yl]-3-(4-fluorophenyl)urea
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	EC50: 0.13 µM (FPR2); 1.1 µM (FPR1)[1]
ln Vitro	FPR2 agonist 2 (compound (S)-11l) (1-100 µM; 48 h) has an EC50 value of 20.8 µM in N9 cells, indicating little cytotoxicity[1]. FPR2 agonist 2 (FPR1/FPR2 HL60 cells) exhibits agonist activity for FPR2 and FPR1, with corresponding EC50s of 0.13 µM and 1.1 µM (IC50s of 0.085 µM, Not determined)[1]. FPR2 agonist 2 (0.1 µM) efficiently decreases the effect of LPS stimulation and inhibits NO generation and LPS-induced cell death[1]. FPR2 agonist 2 (0.1 µM) inhibits caspase-3 activity and balances the alterations in mitochondrial function[1].
ln Vivo	FPR2 agonist 2 has the capacity to cross the blood-brain barrier and build up in the brain (1 mg/kg for intravenous administration; 10 mg/kg for intraperitoneal][1].
Cell Assay	Cell Viability Assay[1] Cell Types: N9 cells Tested Concentrations: 1-100 µM Incubation Duration: 48 h Experimental Results: demonstrated low cytotoxicity with an EC50 value of 20.8 µM in N9 cells.
Animal Protocol	Animal/Disease Models: 25-30 g, male CD-1 mice[1] Doses: Route of Administration: 1 mg/kg for iv; 10 mg/kg for ip (dissolved in 5% DMSO, 10% solutol HS 15, and 85% sterile water) Experimental Results: demonstrated the ability to permeate the blood− brain barrier and to accumulate in the brain.
References	[1]. Mastromarino M, et al. Design, Synthesis, Biological Evaluation, and Computational Studies of Novel Ureidopropanamides as Formyl Peptide Receptor 2 (FPR2) Agonists to Target the Resolution of Inflammation in Central Nervous System Disorders. J Med Chem. 2022; 65(6):5004-5028.

Solubility Data

Solubility (In Vitro)

DMSO: 50 mg/mL (111.99 mM)

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.2399 mL	11.1995 mL	22.3989 mL
	5 mM	0.4480 mL	2.2399 mL	4.4798 mL
	10 mM	0.2240 mL	1.1199 mL	2.2399 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.