INCB3619 (INCB-3619) is a novel, potent, orally bioavailable and highly specific ADAM10 inhibitor, but also markedly inhibits ADAM17, MMP12 and MMP15 with anticancer activity. INCB3619 inhibits the EGFR ligand signaling in the EGFR autocrine cell line NCI-H1666 and sensitizes it to gefitinib. In Min mice, INCB3619 reduced EGFR signals in adenomas and inhibited intestinal tumor multiplicity (P < 0.05). In the AOM model, colonocyte ADAM17 deletion reduced EGFR signals and colonic tumor development (P < 0.05). Finally, ADAM17 was upregulated >2.5-fold in human malignant colonocytes.

Physicochemical Properties

Molecular Weight	413.46688580513
Exact Mass	413.195
CAS #	791826-72-7
PubChem CID	11625778
Appearance	Solid powder
LogP	2.76
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	4
Heavy Atom Count	30
Complexity	726
Defined Atom Stereocenter Count	2
SMILES	O=C([C@@H]1[C@@H](C(N2CC=C(C3C=CC=CC=3)CC2)=O)N(C(=O)OC)CC2(C1)CC2)NO
InChi Key	CKZHFOKQZRZCPF-ROUUACIJSA-N
InChi Code	InChI=1S/C22H27N3O5/c1-30-21(28)25-14-22(9-10-22)13-17(19(26)23-29)18(25)20(27)24-11-7-16(8-12-24)15-5-3-2-4-6-15/h2-7,17-18,29H,8-14H2,1H3,(H,23,26)/t17-,18-/m0/s1
Chemical Name	methyl (5S,6S)-5-(hydroxycarbamoyl)-6-(4-phenyl3,6-dihydro-2H-pyridine-1-carbonyl)-7-azaspiro[2.5]octane-7-carboxylate
Synonyms	INCB3619 INCB-3619 INCB 3619.
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	ADAM10 (IC50 = 22 nM); ADAM17 (IC50 = 12 nM)
ln Vitro	While INCB3619 (0-0.25 μM, 96 h) can prevent HER3-Akt dye from being activated by heregulin, it is unable to stop ERK activity in A549 cells and can cause A549 cytokines [1]. INCB3619 (0-10 μM, 72 h) can be used in conjunction with other anti-EGFR medications, such as gefitinib, to reduce the amount of EGFR ligand signaling in the EGFR cell line NCI-H1666 [1]. NCI-H166 cells exposed to INCB3619 (2 µM) have reduced ERK1/2 expression, rendering them Gefitinib-sensitive [1]. Analysis of apoptosis[1].
ln Vivo	In the BALB/c nu/nu paradigm of A549 xenografts, INCB3619 (subcutaneous injection, 60 mg/kg/d, 14 d) has anti-tumor action, removing tumors that are gefitinib-sensitive [1].
Cell Assay	Apoptosis analysis[1]. Cell Types: A549 Cell Tested Concentrations: 1, 10 μM Incubation Duration: Experimental Results: Approximately 3% cell apoptosis was induced at the 1 μM concentration, and approximately 5% cell apoptosis was induced at the 10 μM concentration. Cell viability assay [1]. Cell Types: NCI-H1666 Cell Tested Concentrations: 0-10 μM Incubation Duration: 72 hrs (hours) Experimental Results: NCI-H1666 cell proliferation was inhibited.
Animal Protocol	Animal/Disease Models: BALB/c nu/nu mouse model of A549 xenograft Doses: 50, 60 mg/kg Route of Administration: subcutaneous injection, daily, 14 days Experimental Results: Significant tumor growth inhibition and delaying effect at 60 mg/kg dose , lower activity. Effect of 50 mg/kg.
References	[1]. Targeting ADAM-mediated ligand cleavage to inhibit HER3 and EGFR pathways in non-small cell lung cancer. Cancer Cell. 2006 Jul;10(1):39-50.

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	2.4186 mL	12.0928 mL	24.1856 mL
	5 mM	0.4837 mL	2.4186 mL	4.8371 mL
	10 mM	0.2419 mL	1.2093 mL	2.4186 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.