HDM201 HCl (also called Siremadlin; NVP-HDM201, HDM stands for 'human double minute 2 homolog') is an orally bioavailable, highly potent and selective inhibitor of the p53-Mdm2 protein-protein interaction that is orally bioavailable. It has a selectivity ratio of more than 1000-fold against Mdm4 and a high affinity for Mdm2 in the picomolar range. A phase I clinical trial using HDM201 is currently being conducted to treat cancer. The HDM2 protein's ability to bind to the p53 tumor suppressor protein's transcriptional activation domain is inhibited by HDM201. The proteosome-mediated enzymatic degradation of p53 is inhibited by preventing this HDM2-p53 protein-protein interaction, which restores both p53 signaling and p53-mediated induction of tumor cell apoptosis.

Physicochemical Properties

Molecular Formula	C26H24CL2N6O4
Molecular Weight	555.41
Exact Mass	554.123
CAS #	1448867-42-2
Related CAS #	Siremadlin;1448867-41-1
PubChem CID	71679278
Appearance	White to off-white solid powder
LogP	3.6
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	38
Complexity	987
Defined Atom Stereocenter Count	1
SMILES	CC(C)N1C2=C(C(=O)N([C@@H]2C3=CC=C(C=C3)Cl)C4=CC(=CN(C4=O)C)Cl)N=C1C5=CN=C(N=C5OC)OC
InChi Key	AGBSXNCBIWWLHD-HXUWFJFHSA-N
InChi Code	InChI=1S/C26H24Cl2N6O4/c1-13(2)33-21-19(30-22(33)17-11-29-26(38-5)31-23(17)37-4)25(36)34(18-10-16(28)12-32(3)24(18)35)20(21)14-6-8-15(27)9-7-14/h6-13,20H,1-5H3/t20-/m1/s1
Chemical Name	(4R)-5-(5-chloro-1-methyl-2-oxopyridin-3-yl)-4-(4-chlorophenyl)-2-(2,4-dimethoxypyrimidin-5-yl)-3-propan-2-yl-4H-pyrrolo[3,4-d]imidazol-6-one
Synonyms	HDM-201; HDM 201; HDM201; NVP-HDM 201; NVP-HDM201; NVP-HDM-201
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	In vitro activity: HDM201 acts by binding to the p53 binding-site of the Mdm2 protein, thereby disrupting the interaction of the two proteins (p53 and Mdm2) and leading to the activation of the p53 pathway. In human tumor cells with p53 wild-type, it causes a robust cell cycle arrest and apoptosis. A panel of cancer cell lines that HDM201 exhibits high selectivity toward.
ln Vivo	Treatment was initiated when the tumors engrafted in the flank were at least 150 mm3. The enrollment process was random. With the measurements of tumor volumes at the beginning of treatment, efficacy studies, tumor response, and relapse were reported. Two times per week, at intervals of 3 days and 4 days alternately, HDM201 was given orally at a dose of 100 mg/kg in a solution of 0.5% methylcellulose and 0.1% Tween 80. Vehicle was produced using the formulation. Early in the morning, treatments were given.
Cell Assay	The in vitro combination screen was carried out on cancer cell lines at Horizon Discovery (Cambridge, MA), and data analysis was done as previously mentioned. Here, we concentrated our data analysis on combinations involving CGM097, a member of an earlier generation of selective TP53-MDM2 inhibitors. Various concentrations of CGM097 and 25 additional compounds were applied to a total of 485 cancer cell lines. We combined the data on TP53 mutation status and distinguished between cell lines that did not have TP53 mutations and cell lines that did. We evaluated the synergistic effects of combining HDM201 and A-1155463 as previously mentioned.
Animal Protocol	HDM201 was administered at 100 mg/kg in 0.5% methylcellulose and 0.1% Tween 80 orally twice a week Mouse
References	[1]. Bioorg Med Chem Lett. 2016 Oct 1;26(19):4837-4841.

Solubility Data

Solubility (In Vitro)

DMSO: ~100 mg/mL (~180.0 mM) Water: < 1mg/mL Ethanol: ~3 mg/mL (~5.4 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	1.8005 mL	9.0024 mL	18.0047 mL
	5 mM	0.3601 mL	1.8005 mL	3.6009 mL
	10 mM	0.1800 mL	0.9002 mL	1.8005 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.