LasB-IN-1 (compound 5f) is a highly potent and orally effective LasB inhibitor (IC50 = 8.7 μM). LasB-IN-1 can effectively inhibit the production of elastase and biofilm formation by Pseudomonas aeruginosa, while reducing inflammatory responses by downregulating MAPK and NF-κB pathways. LasB-IN-1 may become a new candidate agent for anti-resistant infections.

Physicochemical Properties

Molecular Formula	C20H22F3NO2
Molecular Weight	365.39
Appearance	Typically exists as solid at room temperature
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	IC50: 8.7 μM (LasB)[1]
ln Vitro	LasB-IN-1 (5-10 μM, 10 h) dose-dependently inhibits the expression of LasB (IC50 = 8.7 μM), elastase biosynthesis (IC50 = 7.3 μM) and biosynthesis in Pseudomonas aeruginosa PAO1 strain. Film formation (IC50 = 7.4 μM) [1]. LasB-IN-1 (1.25-20 μM, 24 h) has no cytotoxic effect on RAW264.7 cells and Vero cells. LasB-IN-1 (50-100 μM, 24 h) exhibits mild cytotoxicity. LasB-IN-1 (0.625-10 μM, 4 h) has negligible hemolytic effects on mouse and human erythrocytes [1]. LasB-IN-1 (10 μM, 4, 6 h) can effectively inhibit the migration of zebrafish larval macrophages to the injury site [1]. LasB-IN-1 (0.625-10 μM, 27 h) inhibits the production of IL-1β, TNF-α, and IL-6 in RAW264.7 cells treated with LPS (1 μg/mL, 24 h). LasB-IN-1 (1.25-10 μM, 27 h) can also inhibit the mRNA expression of COX-2, iNOS, IL-1β, TNF-α and IL-6 [1]. LasB-IN-1 (1.25-10 μM, 27 h) significantly inhibited the phosphorylation of NF-κB p65, i-κB α, JNK and ERK in RAW264.7 cells treated with LPS (1 μg/mL, 24 h). And it is concentration-dependent [1].
ln Vivo	LasB-IN-1 (1, 2 mg/kg, gavage, 4 h) inhibited neutrophil infiltration in mouse lung tissue[1]. LasB-IN-1 (1, 2 mg/kg, gavage, 4 h) significantly downregulated the expression levels of IL-1β, TNF-α and IL-6 in mice, and reduced the expression of COX-2, iNOS, IL-1β, TNF-α and IL-6 mRNA[1]. LasB-IN-1 (2, 4 mg/kg, gavage, once a day for 7 days) had no obvious adverse reactions[1].
Cell Assay	Cell Viability Assay[1] Cell Types: RAW264.7 cell and Vero cell Tested Concentrations: 1.25, 2.5, 5, 10 , 20, 50, 100 μM Incubation Duration: 24 h Experimental Results: Revealed no cytotoxic effect at concentrations of lower than 20 μM, a slight cytotoxic effect was observed at a concentration of 50 μM, indicating the noncytotoxicity at concentrations effective against virulence and biofilm formation .
Animal Protocol	Animal/Disease Models: eight-week-old BALB/c wild-type mice (male, 18–22 g ), established a ALI model by using LPS (P. aeruginosa)[1] Doses: 1 and 2 mg/kg Route of Administration: intragastric (i.g.), after treatment for 4 h, intraperitoneal injection of LPS (20 mg/kg), sacrificed by cervical dislocation after a 6 h treatment Experimental Results: Reduced myeloperoxidase (MPO) activity in mice compared with that in LPS-only mice, indicating the inhibition of neutrophil infiltration in mouse lung tissues. Exhibited significantly downregulated expression levels of IL-1β, TNF-α, and IL-6, with a concentration-dependent response while significantly reduced mRNA expression of COX-2, iNOS, IL-1β, TNF-α, and IL-6 in mice compared with LPS-only mice. Animal/Disease Models: eight-week-old BALB/c wild-type mice (male, 18–22 g )[1] Doses: 2 and 4 mg/kg Route of Administration: intragastric (i.g.) for 7 days Experimental Results: Exhibited no adverse effects such as vomiting or diarrhea, had a negligible impact on the body weight of mice, did not induce any significant damage or histopathological alterations in vital organs.
References	[1].Novel ligustilide derivatives target quorum sensing system LasR/LasB and relieve inflammatory response against Pseudomonas aeruginosa infection. European Journal of Medicinal Chemistry. 2023，263，115972.

Solubility Data

Solubility (In Vitro)

Typically soluble in DMSO (e.g. 10 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	2.7368 mL	13.6840 mL	27.3680 mL
	5 mM	0.5474 mL	2.7368 mL	5.4736 mL
	10 mM	0.2737 mL	1.3684 mL	2.7368 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.