Vevorisertib (ARQ 751) triHCl is a selective, allosteric, pan-AKT and AKT1-E17K mutant inhibitor that potently inhibits AKT phosphorylation. The Kds of Vevorisertib triHCl for AKT1 and AKT1-E17K are 1.2 nM and 8.6 nM, respectively. Vevorisertib triHCl has IC50s of 0.55, 0.81 and 1.3 nM for AKT1, AKT2 and AKT3 respectively. Vevorisertib triHCl may be used in cancer-related research.

Physicochemical Properties

Molecular Formula	C35H41CL3N8O
Molecular Weight	696.11
Exact Mass	694.246
CAS #	1416775-08-0
Related CAS #	Vevorisertib;1416775-46-6
PubChem CID	131746460
Appearance	White to yellow solid powder
Hydrogen Bond Donor Count	5
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	47
Complexity	976
Defined Atom Stereocenter Count	0
SMILES	NC1N=CC=CC=1C1=NC2=CC=C(C3C=CC=C(N4CCC(N(C)C(=O)C)CC4)C=3)N=C2N1C1C=CC(C2(CCC2)N)=CC=1.Cl.Cl.Cl
InChi Key	HNFNJTGXAZHZSB-UHFFFAOYSA-N
InChi Code	InChI=1S/C35H38N8O.3ClH/c1-23(44)41(2)26-15-20-42(21-16-26)28-7-3-6-24(22-28)30-13-14-31-34(39-30)43(33(40-31)29-8-4-19-38-32(29)36)27-11-9-25(10-12-27)35(37)17-5-18-35;;;/h3-4,6-14,19,22,26H,5,15-18,20-21,37H2,1-2H3,(H2,36,38);3*1H
Chemical Name	N-[1-[3-[3-[4-(1-aminocyclobutyl)phenyl]-2-(2-aminopyridin-3-yl)imidazo[4,5-b]pyridin-5-yl]phenyl]piperidin-4-yl]-N-methylacetamide;trihydrochloride
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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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	Inhibiting AKT1-E17K phosphorylation, vevorisertib triHClide (0, 12, 33, 111, 333, 1000 nM, 2 hours) [1]. AKT-WT and AKT1-E17K plasma membrane translocation is inhibited by vevorisertib triHClide (1 μM) for two hours in NIH 3T3 cells transfected with pcDNAAKT-WT-GFP or pcDNA-E17K-GFP, whether growth factors are present or not [1]. Full-length AKT1 is 57% inhibited by vevorisertib triHCle (5 μM) [1]. Vevorisertib triHClide (0, 0.012, 0.037, 0.11, 0.33, 1 μM; 2 hours) affects AKT direct substrates, such as PRAS40, GSK3β, FOXO, BAD, and AS160, and mTORC1 in cancer cell lines in a dose-dependent manner [1]. Vevorisertib triHClide (GI50 < 1 μM) inhibits the growth of cancer cells in the head and neck, breast, and esophageal regions [1]. In PIK3CA mutant cell lines, vevorisertib triHClide demonstrates strong antiproliferative action [1]. In gastrointestinal stromal tumor (GIST) cells, vevorisertib triHCl (MK-4440)/imatinib mesylate (IM) combination exhibits cell cycle arrest and enhanced cell death [2]. With PIK3CA mutant cell lines, vevorisertib triHClide demonstrates potent antiproliferative activity [1]: GI50 (nM) PIK3CA ER PR HER2 T47D 1.05 H1047R + + - EFM-19 1.54 H1047R + + - MCF-7 2.20 E545K + + - BT474 3.25 K111N + + + MDA-MB-453 6.05 H1047R - - +
ln Vivo	Vevorisertib triHClide (25, 50 and 75 mg/kg; oral; 5 days on and then 4 days off for 20 days) indicated effective tumor growth suppression of 68%, 78% and 98%, respectively [1]. Vevorisertib triHClide (5, 10, 20, 40, 80 and 120 mg/kg; given orally daily for ten days) exhibited tumor growth inhibition of 29%, 33%, 50%, 73%, 83% and 92%, respectively. [1]. Vevorisertib triHCl has a Cmax plasma concentration of ≥2 μM [1]. Vevorisertib triHClide is generally well tolerated at dose levels up to 120 mg/kg [1]. Compared with either single drug, the Vevorisertib triHClide (MK-4440)/IM combination showed greater efficacy in preclinical models of IM-sensitive GIST [2].
Cell Assay	Western Blot Analysis[1] Cell Types: 293T cells (transiently transfected with pcDNA-E17K-GFP) Tested Concentrations: 0, 12, 33, 111, 333. 1000 nM Incubation Duration: 2 hrs (hours) Experimental Results: AKT1-E17K phosphorylation is inhibited. Western Blot Analysis[1] Cell Types: Cancer Cell Types: MDA-MB 453 (PIK3CAH1047R; Her2 amp), NCI-H1650 (PTEN null), KU-19-19 (AKT1-E17K&E49K; NRas Q61R) Tested Concentrations: 0, 0.012 , 0.037, 0.11, 0.33, 1 μM Incubation Duration: 2 hrs (hours) Experimental Results: Displayed dose-dependent effects on mTORC1 and AKT direct substrates including PRAS40, GSK3β, FOXO, BAD and AS160.
Animal Protocol	Animal/Disease Models: Endometrial PDX mouse xenograft model (AKT1-E17K mutant tumor fragments are subcutaneously (sc) (sc) implanted into athymic nude mice; tumor volume is approximately 200 mm3) [1] Doses: 25, 50 and 75 mg/kg Route of Administration: po; 5 days of dosing, followed by a 4-day dosing holiday, for a total of 20 days. Experimental Results: demonstrated effective tumor growth inhibition of 68%, 78% and 98% respectively. Animal/Disease Models: AN3CA mouse xenograft model (female NCr nu/nu mouse with tumor size 250 mm3) [1] Doses: 5, 10, 20, 40, 80 and 120 mg/kg Route of Administration: Oral; daily Once, the results lasted for ten days: showing tumor growth inhibition of 29%, 33%, 50%, 73%, 83% and 92% respectively.
References	[1]. Targeting AKT1-E17K and the PI3K/AKT Pathway with an Allosteric AKT Inhibitor, ARQ 092. PLoS One. 2015 Oct 15;10(10):e0140479. [2]. Combined Inhibition of AKT and KIT Restores Expression of Programmed Cell Death 4 (PDCD4) in Gastrointestinal Stromal Tumor. Cancers (Basel). 2021 Jul 23;13(15):3699.

Solubility Data

Solubility (In Vitro)

DMSO : ~150 mg/mL (~215.48 mM) H2O : ~25 mg/mL (~35.91 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 5.25 mg/mL (7.54 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 52.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 5.25 mg/mL (7.54 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 52.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.4366 mL	7.1828 mL	14.3655 mL
	5 mM	0.2873 mL	1.4366 mL	2.8731 mL
	10 mM	0.1437 mL	0.7183 mL	1.4366 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.