Zunsemetinib (ATI-450; CDD-450) is a novel and potent inhibitor of p38α mitogen-activated protein kinase-activated protein kinase 2 (MK2) with the potential to be used for immuno-inflammatory disease.

Physicochemical Properties

Molecular Formula	C25H22CLF2N5O3
Molecular Weight	513.923691272736
Exact Mass	513.137
Elemental Analysis	C, 58.43; H, 4.31; Cl, 6.90; F, 7.39; N, 13.63; O, 9.34
CAS #	1640282-42-3
Related CAS #	(R)-Zunsemetinib;1640282-44-5
PubChem CID	86291496
Appearance	White to yellow solid powder
LogP	2.5
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	6
Heavy Atom Count	36
Complexity	888
Defined Atom Stereocenter Count	0
InChi Key	FQPQMJULRZINPV-UHFFFAOYSA-N
InChi Code	InChI=1S/C25H22ClF2N5O3/c1-13-10-30-18(17-5-6-29-24(32-17)25(3,4)35)9-20(13)33-14(2)7-21(22(26)23(33)34)36-12-19-16(28)8-15(27)11-31-19/h5-11,35H,12H2,1-4H3
Chemical Name	(2'S)-3-Chloro-4-[(3,5-difluoro-2-pyridinyl)methoxy]-2'-[2-(1-hydroxy-1-methylethyl)-4-pyrimidinyl]-5',6-dimethyl[1(2H),4'-bipyridin]-2-one
Synonyms	(P)-ATI-450 ATI-450CDD-450ATI450CDD450ATI 450 CDD 450ZunsemetinibATI-450 P atropisomer
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	While zunsemetinib (1 and 10 μM; 1 hour; WT and NOM ID BMM) has no effect on NLRP3 expression, it promotes the degradation of IL-1β mRNA, which lowers the expression of IL-1β [1]. Zunsemetinib (0.4 nM~1 μM; 16 hours; PBMC) increases the instability of IL-1β mRNA and decreases the production of IL-1β [1]. Zunsemetinib preserves the p38α activation of other effectors including PRAK and ATF2, but specifically prevents the proinflammatory kinase MK2 from being activated by p38α MAPK. In vitro, RANKL-induced osteoclast production is inhibited by zunsemetinib [1].
ln Vivo	Zunsemetinib (1,000 ppm; oral) inhibits LPS-induced TNF-α expression for up to 4 weeks after dosing [1]. Zunsemetinib (10 and 20 mg/kg; oral) enhances bone mineral density [1]. Zunsemetinib reduces osteopenia in NOM IDc mice by suppressing osteoclastogenesis [1].
Cell Assay	RT-PCR[1] Cell Types: WT and NOM ID BMM Tested Concentrations: 1 and 10 μM Incubation Duration: 1 hour Experimental Results: No effect on NLRP3 expression, but diminished IL-1β expression by promoting IL-1β mRNA degradation.
Animal Protocol	Animal/Disease Models: 8weeks old WT female mice [1] Doses: 1,000 ppm Route of Administration: Oral Experimental Results: LPS-induced blockade of TNF-α expression persisted for up to 4 weeks after administration. Animal/Disease Models: Rat[1] Doses: 10 and 20 mg/kg Route of Administration: Oral Experimental Results:Increased bone density.
References	[1]. Zunsemetinib (ATI-450) – Investigational oral MK2 pathway inhibitor. [2]. Aclaris Therapeutics Announces ATI-450 (MK2 pathway Inhibitor) publication in Journal of Experimental Medicine. [3]. Selective inhibition of the p38α MAPK-MK2 axis inhibits inflammatory cues including inflammasome priming signals. J Exp Med. 2018;215(5):1315-1325.
Additional Infomation	Zunsemetinib is an orally bioavailable, small molecule inhibitor of mitogen-activated protein (MAP) kinase-activated protein kinase 2 (MAPKAPK2; MK2), with potential anti-inflammatory activity. Upon oral administration, zunsemetinib targets and binds to the p38MAPK-MK2 complex, thereby inhibiting the p38MAPK phosphorylation and activation of MK2. This inhibits p38MAPK/MK2-mediated inflammatory signaling pathway. This may result in the inhibition of the production of proinflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1-alpha, IL-1-beta and IL-6.

Solubility Data

Solubility (In Vitro)

DMSO : ~100 mg/mL (~194.58 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	1.9458 mL	9.7291 mL	19.4583 mL
	5 mM	0.3892 mL	1.9458 mL	3.8917 mL
	10 mM	0.1946 mL	0.9729 mL	1.9458 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.