NITD-349 is a novel and potent MmpL3 inhibitor, it shows highly potent anti-mycobacterial activity (MIC50=23 nM) against virulent Mycobacterium tuberculosis H37Rv.

Physicochemical Properties

Molecular Formula	C17H20F2N2O
Molecular Weight	306.350311279297
CAS #	1473450-62-2
PubChem CID	72711190
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	2
Heavy Atom Count	22
Complexity	419
Defined Atom Stereocenter Count	0
InChi Key	UATYSFRIVIHVKL-UHFFFAOYSA-N
InChi Code	InChI=1S/C17H20F2N2O/c1-17(2)5-3-11(4-6-17)20-16(22)15-9-12-13(19)7-10(18)8-14(12)21-15/h7-9,11,21H,3-6H2,1-2H3,(H,20,22)
Chemical Name	N-(4,4-dimethylcyclohexyl)-4,6-difluoro-1H-indole-2-carboxamide
Synonyms	NITD-349NITD 349 NITD349
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

ln Vitro	Mycobacterium tuberculosis (Mtb) reproducing in vitro is inhibited by NITD-349, which also possesses bactericidal effect against Mtb within macrophages. Upon analyzing the killing kinetics of these compounds, it was shown that the decrease of Mtb cells was dependent on both concentration and duration, resulting in a reduction of 3 to 4 logarithmic units (CFU) within 3 days of treatment. NITD-304's lethal activity features are comparable to those of isoniazid, which can kill quickly at concentrations higher than 0.2 μM. NITD349 exhibited MIC activity ranging from 0.04 to 0.08 μM against multiple MDR Mtb strains. In vitro metabolic clearance of NITD-349 in mouse and human liver microsomes is moderate and it demonstrates high permeability [1].
ln Vivo	NITD-349 was efficacious in mouse models of both acute and chronic Mycobacterium TB infection, and it showed positive oral pharmacokinetic (PK) qualities in dogs and rodents in acute mice effectiveness models. Treatment of mice with NITD-349 at dosages of 12.5 and 50 mg/kg led to a reduction of 0.9 and 3.4 logarithmic units in lung tissue CFU in an acute animal efficacy model. After two weeks of treatment, NITD-349 outperformed ethambutol and was as effective as rifampicin, the first-line medication for tuberculosis, in known animal models of infection. CFU decreased by 2.38 logs during a 4-week course of treatment with NITD-349 at a dose of 100 mg/kg [1].
References	[1]. Rao SP, et al. Indolcarboxamide is a preclinical candidate for treating multidrug-resistant tuberculosis. Sci Transl Med. 2013 Dec 4;5(214):214ra168

Solubility Data

Solubility (In Vitro)

DMSO : ≥ 310 mg/mL (~1011.91 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	3.2642 mL	16.3212 mL	32.6424 mL
	5 mM	0.6528 mL	3.2642 mL	6.5285 mL
	10 mM	0.3264 mL	1.6321 mL	3.2642 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.