OD36 HCl is a RIPK2 inhibitor (antagonist) with IC50 of 5.3 nM. OD36 HCl is a macrocyclic inhibitor that binds efficiently to the ALK2 kinase ATP pocket. OD36 HCl targets ALK2 with Kd of 37 nM.

Physicochemical Properties

Molecular Formula	C16H16CL2N4O2
Molecular Weight	367.2298412323
Exact Mass	366.065
CAS #	2387510-88-3
Related CAS #	OD36;1638644-62-8
PubChem CID	138991767
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	0
Heavy Atom Count	24
Complexity	402
Defined Atom Stereocenter Count	0
InChi Key	VGOJZWODJXQKMN-UHFFFAOYSA-N
InChi Code	InChI=1S/C16H15ClN4O2.ClH/c17-12-7-11-8-13(9-12)23-6-5-22-4-2-18-15-1-3-21-16(20-15)14(11)10-19-21;/h1,3,7-10H,2,4-6H2,(H,18,20);1H
Chemical Name	4-chloro-7,10-dioxa-13,17,18,21-tetrazatetracyclo[12.5.2.12,6.017,20]docosa-1(20),2(22),3,5,14(21),15,18-heptaene;hydrochloride
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	Additionally, OD36 inhibits ALK2 and ALK2 R206H, with IC50 values of 47 and 22 nM, respectively[1]. When ALK1 is exposed to OD36, 90 nM is the KD [2]. ALK2 signaling mutants and osteogenic differentiation are successfully inhibited by OD36 [2]. BMP-6 (50 ng/mL)-induced p-Smad1/5 in KS483 cells is efficiently inhibited by OD36 (0.1-1 μM; 24 hours) [2]. The preincubation of FOP endothelial colony-forming cells (ECFC) with OD36 (0.5 μM) totally inhibits the activation of Smad1/5, which is responsible for activin A-induced gene targets ID-1 and ID-3 [2].
ln Vivo	In a mouse model of acute peritonitis, OD36 (6.25 mg/kg; intraperitoneal injection; once) reduces inflammation [3].
Cell Assay	Western Blot Analysis[2] Cell Types: KS483 Cell Tested Concentrations: 0.1, 0.2 and 1 μM Incubation Duration: 24 hrs (hours) Experimental Results: Inhibition of BMP-6 induced p-Smad1/5.
Animal Protocol	Animal/Disease Models: C57BL/6 mice, muramyl dipeptide (MDP)-induced peritonitis model [3] Doses: 6.25 mg/kg Route of Administration: intraperitoneal (ip) injection, 30 minutes before MDP Experimental Results: Inhibited the influx of inflammatory cells into the peritoneum Recruits, particularly neutrophils, and to a lesser extent lymphocytes. Expression of RIPK2-specific genes as well as inflammatory cytokine and chemokine genes was diminished.
References	[1]. In vivo inhibition of RIPK2 kinase alleviates inflammatory disease. J Biol Chem. 2014 Oct 24;289(43):29651-64. [2]. Development of Macrocycle Kinase Inhibitors for ALK2 Using Fibrodysplasia Ossificans Progressiva-Derived Endothelial Cells. JBMR Plus. 2019 Oct 7;3(11):e10230. [3]. In vivo inhibition of RIPK2 kinase alleviates inflammatory disease. J Biol Chem. 2014 Oct 24;289(43):29651-64.

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	2.7231 mL	13.6154 mL	27.2309 mL
	5 mM	0.5446 mL	2.7231 mL	5.4462 mL
	10 mM	0.2723 mL	1.3615 mL	2.7231 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.