mTOR/HDAC-IN-1 (Compound 50) is a selective dual (bifunctional) inhibitor of mTOR and HDAC, with IC50 of 0.49 and 0.91 nM for mTOR and HDAC1, respectively. mTOR/HDAC-IN-1 can be studied as an anti-cancer active molecule.

Physicochemical Properties

Molecular Formula	C23H23N11O3
Molecular Weight	501.50
Exact Mass	501.198
CAS #	2815286-02-1
PubChem CID	163321997
Appearance	Typically exists as solid at room temperature
LogP	0.9
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	12
Rotatable Bond Count	5
Heavy Atom Count	37
Complexity	794
Defined Atom Stereocenter Count	0
InChi Key	VPTVCYOEECUFBP-UHFFFAOYSA-N
InChi Code	InChI=1S/C23H23N11O3/c24-19-17-18(13-1-2-16-15(7-13)30-22(25)37-16)31-34(20(17)29-11-28-19)10-12-3-5-33(6-4-12)23-26-8-14(9-27-23)21(35)32-36/h1-2,7-9,11-12,36H,3-6,10H2,(H2,25,30)(H,32,35)(H2,24,28,29)
Chemical Name	2-[4-[[4-amino-3-(2-amino-1,3-benzoxazol-5-yl)pyrazolo[3,4-d]pyrimidin-1-yl]methyl]piperidin-1-yl]-N-hydroxypyrimidine-5-carboxamide
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	mTOR 0.49 nM (IC50) HDAC1 0.91 nM (IC50) HDAC6 86 nM (IC50) HDAC8 27 nM (IC50)
ln Vitro	Against A549, HCT116, and MV4-11 cells, mTOR/HDAC-IN-1 (Compound 50) (0-10 μM, 72 h) has strong anti-proliferative activity[1]. At the cellular level, mTOR/HDAC-IN-1 (0–3 μM, 6 h) concurrently regulates HDAC catalytic activity and mTOR signaling[1]. With IC50 values of 0.91, 86, 27, and >1000 against HDAC1, HDAC6, HDAC8, and HDAC11, mTOR/HDAC-IN-1 exhibits remarkable specificity over both HDAC6 and HDAC11 [1]. MLN0128's connection with mTOR's ATP-binding site is maintained by mTOR/HDAC-IN-1, which also binds to HDAC's catalytic channel[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: A549, HCT116, and MV4-11 Tested Concentrations: 0-10 μM Incubation Duration:At 70 h post-compound treatment, to each well was added CCK-8, and the mixture was incubated for additional 2 h. Experimental Results: demonstrated anti-proliferative activity with IC50 values of 2.16, 3.85, and 1.74 μM against A549, HCT116, and MV4-11 cells, respectively. Western Blot Analysis[1] Cell Types: MV4-11 Tested Concentrations: 0.3, 1.0, and 3.0 μM Incubation Duration: 6 h Experimental Results: Downregulated the levels of Phos-S6 (Ser235/Ser236) and Phos-AKT (Ser473), and elevated the levels of Ac-H3 (K9) and Acα-tubulin (Lys40) in a concentration-dependent manner.
References	[1]. Discovery of novel pyrazolopyrimidine derivatives as potent mTOR/HDAC bi-functional inhibitors via pharmacophore-merging strategy. Bioorg Med Chem Lett. 2021 Oct 1;49:128286.

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	1.9940 mL	9.9701 mL	19.9402 mL
	5 mM	0.3988 mL	1.9940 mL	3.9880 mL
	10 mM	0.1994 mL	0.9970 mL	1.9940 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.