Mps1-IN-5 is a potent, orally bioactive Mps1 inhibitor (antagonist) with IC50 of 29 nM. Mps1-IN-5 causes apoptosis and cell cycle arrest in the G2/M phase. Mps1-IN-5 displays antiproliferation and anticancer effect. Mps1-IN-5 inhibits the phosphorylation of Mps1 and induces DNA damage.

Physicochemical Properties

Molecular Formula	C24H25N9
Molecular Weight	439.52
CAS #	2890819-31-3
Appearance	Typically exists as solid at room temperature
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	IC50: 29 nM (Mps1)[1]
ln Vitro	Compound 12 (Mps1-IN-5) (0-10 µM, 24, 48, 72 h) blocks MCF-7 and 4T1 cell proliferation in a time-dependent way[1]. Mps1-IN-5 (0, 0.5, 1.0, 5.0 µM; 24, 48 h) induces apoptosis and cell cycle arrest in the G2/M phase in MCF-7 and 4T1 cells in a dose-dependent way[1]. Mps1-IN-5 (0, 0.03, 0.1, 1, 3 µM; 2 h) causes DNA damage and inhibits Mps1 phosphorylation[1].
ln Vivo	In breast cancer models, Mps1-IN-5 (30, 60 mg/kg; po; daily for 15 days) reduces tumor growth without clearly visible damage [1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: MDA-MB-231, MCF-7, 4T1, HEY, OVCAR- 3, ES-2, HCT-116, A549, AGS cells Tested Concentrations: 0-3 µM Incubation Duration: 72 h Experimental Results: Inhibited the cell growth with IC50s of 2.68, 0.37, 0.40, >3, >3, >3, 1.06, 2.03, >3 µM for MDA-MB-231, MCF-7, 4T1, HEY, OVCAR-3, ES-2, HCT-116, A549, AGS cells, respectively. Cell Cycle Analysis[1] Cell Types: MCF -7, 4T1 cells Tested Concentrations: 0, 0.5, 1.0, 5.0 µM Incubation Duration: 24 h Experimental Results: Induced cell cycle arrest of MCF-7 and 4T1 cells at the G2/M phase in a dose-dependent manner, diminished the protein expression levels of Cyclin B1 and CDK1. Apoptosis Analysis[1] Cell Types: MCF-7, 4T1 cells Tested Concentrations: 0, 0.5, 1.0, 5.0 µM Incubation Duration: 48 h Experimental Results: Induced apoptosis and Dramatically increased the expression level of an apoptosis- related protein, cleaved poly ADP-ribose polymerase (PARP). Western Blot Analysis[1] Cell Types: MCF-7, 4T1 cells Tested Concentrations: 0, 0.03, 0.1, 1, 3 µM Incubation Duration: 2 h Experimental Results: Increased the expression of level of γ-H2AX protein and diminished the protein expression of p-Mps1 in a dose-dependent manner.
Animal Protocol	Animal/Disease Models: 6-8 weeks, female balb/c (Bagg ALBino) mouse: (4T1-luc mouse xenograft model)[1] Doses: 30, 60 mg/kg Route of Administration: Po; daily for 15 days Experimental Results: Dramatically suppressed tumor growth and caused negligible damage to organs such as heart, liver, spleen, lung and kidneys.
References	[1]. Design, synthesis and biological evaluation of a new class of 7H-pyrrolo[2,3-d]pyrimidine derivatives as Mps1 inhibitors for the treatment of breast cancer. Eur J Med Chem. 2023 Jan 5;245(Pt 1):114887.

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	2.2752 mL	11.3760 mL	22.7521 mL
	5 mM	0.4550 mL	2.2752 mL	4.5504 mL
	10 mM	0.2275 mL	1.1376 mL	2.2752 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.