LeuRS-IN-1 is a potent orally bioactive Mycobacterium tuberculosis leucyl-tRNA synthetase (M.tb LeuRS) inhibitor. The IC50 and Kds of LeuRS-IN-1 against M.tb LeuRS are 0.06 μM and 0.075 μM, respectively. LeuRS-IN-1 inhibits human cytoplasmic LeuRS (IC50=38.8 μM) and HepG2 protein synthesis (EC50=19.6 μM).

Physicochemical Properties

Molecular Formula	C10H13BCLNO3
Molecular Weight	241.479121923447
Exact Mass	241.067
CAS #	1364914-72-6
Related CAS #	LeuRS-IN-1 hydrochloride;1364683-67-9
PubChem CID	56944652
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	3
Heavy Atom Count	16
Complexity	246
Defined Atom Stereocenter Count	1
SMILES	B1(C2=C(C=CC(=C2[C@H](O1)CN)Cl)OCC)O
InChi Key	FAIWKHSRNFCJGO-MRVPVSSYSA-N
InChi Code	InChI=1S/C10H13BClNO3/c1-2-15-7-4-3-6(12)9-8(5-13)16-11(14)10(7)9/h3-4,8,14H,2,5,13H2,1H3/t8-/m1/s1
Chemical Name	[(3S)-4-chloro-7-ethoxy-1-hydroxy-3H-2,1-benzoxaborol-3-yl]methanamine
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	M.tb LeuRS 0.06 μM (IC50) M.tb LeuRS 0.075 μM (Kd) human cytoplasmic LeuRS 38.8 μM (IC50) HepG2 protein synthesis 19.6 μM (EC50)
ln Vitro	LeuRS-IN-1 (compound 13) has a minimum inhibitory concentration (MIC) of 0.02 μg/mL against M.tb H37Rv bacterium [1]. Compound 3a, LeuRS-IN-1, has an EC50 value of 65.8 μM and can cause toxicity in HepG2 cells in 48 hours[2].
ln Vivo	LeuRS-IN-1 (daily oral dose of 100 mg/kg for 14 days) lowers the lung CFU value in mice with acute tuberculosis (TB)[1]. LeuRS-IN-1 (33 mg/kg; taken once daily for four weeks) lowers the CFU values for the lungs and spleen in chronic TB mice[1]. Pharmacokinetic parameters in mice [1]: Management Dosage (g/kg) CL (ml/h/kg) at 5 min Cmax (μg/ml) Vss (ml/kg) MRT in hours AUC0-∞ (h · μg/ml) α-t1/2 (h) (% AUC) β-t1/2 (h) (% AUC) Section IV: 13.6 582 3,142 5.4 51.6 0.10 ( 2) 3.83 (98) Management Dosage (g/kg) Cmax in μg/ml Tmax in hours AUC0-24 (h · μg/ml) Terminal h = t1/2 Bioavailability percentage (h · μg/ml) Mousing PPB (%) 6.4 0.25 47.5 3.1 9.2 23 po 30
Animal Protocol	Animal/Disease Models: Murine GKO (C57BL/6-Ifngtm1ts) model of acute TB[1] Doses: 100 mg/kg Route of Administration: Orally daily for 14 days after 10 days of infection (start) with M. tuberculosis Erdman. Experimental Results: diminished lung CFU value in mice. Animal/Disease Models: Murine BALB/c model of chronic TB infection[1] Doses: 33 mg/kg Route of Administration: Orally 5 days a week for 4 weeks after infection with M. tuberculosis Erdman with a low-dose aerosol 21 days prior (start). Experimental Results: diminished lung and spleen CFU values in mice.
References	[1]. Palencia A, et al. Discovery of Novel Oral Protein Synthesis Inhibitors of Mycobacterium tuberculosis That Target Leucyl-tRNA Synthetase. Antimicrob Agents Chemother. 2016;60(10):6271-6280. Published 2016 Sep 23. [2]. Li X, et al. Discovery of a Potent and Specific M. tuberculosis Leucyl-tRNA Synthetase Inhibitor: (S)-3-(Aminomethyl)-4-chloro-7-(2-hydroxyethoxy)benzo[c][1,2]oxaborol-1(3H)-ol (GSK656). J Med Chem. 2017 Oct 12;60(19):8011-8026.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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	4.1411 mL	20.7056 mL	41.4113 mL
	5 mM	0.8282 mL	4.1411 mL	8.2823 mL
	10 mM	0.4141 mL	2.0706 mL	4.1411 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.