ALM301 is an orally bioactive, specific AKT inhibitor (antagonist) with IC50s of 0.13 μM, 0.09 μM, and 2.75 μM for AKT1, AKT2, and AKT3, respectively. ALM301 inhibits AKT phosphorylation and regulates downstream signaling in vitro. ALM301 inhibits cancer/tumor cell growth/proliferation and tumor growth.

Physicochemical Properties

Molecular Formula	C25H25N3O3
Molecular Weight	415.48
Exact Mass	415.189
CAS #	1313439-71-2
PubChem CID	68299673
Appearance	Off-white to light yellow solid powder
LogP	2.6
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	4
Heavy Atom Count	31
Complexity	638
Defined Atom Stereocenter Count	0
SMILES	O1CC(=O)N(CC)C2=CC(C3=CC=CC=C3)=C(C3=CC=C([C@@]4(N)C[C@H](O)C4)C=C3)N=C12
InChi Key	BPHCAPAOSHZYJY-UHFFFAOYSA-N
InChi Code	InChI=1S/C25H25N3O3/c1-2-28-21-12-20(16-6-4-3-5-7-16)23(27-24(21)31-15-22(28)30)17-8-10-18(11-9-17)25(26)13-19(29)14-25/h3-12,19,29H,2,13-15,26H2,1H3
Chemical Name	6-[4-(1-amino-3-hydroxycyclobutyl)phenyl]-1-ethyl-7-phenylpyrido[2,3-b][1,4]oxazin-2-one
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	ALM301 (0.001-10 µM; 72 h) raises the sub-G0 population in a dose-dependent manner, inhibiting the growth of cancer cells, with PI3KCA mutant MCF-7 cells being the most sensitive [1]. With an EC50 value of 0.47 µM, ALM301 (1 µM; 1 hour) suppresses AKT phosphorylation in MCF-7 for as long as 48 hours [1].
ln Vivo	ALM301 (10, 30 and 100 mg/kg; oral; single dosage) suppresses pAKTS473 in cancers and decreases tumor growth [1]. ALM301 (3 or 10 mg/kg; oral; once daily for 49 days) has showed increased tumor suppressive capabilities when paired with tamoxifen [1].
Cell Assay	Cell proliferation experiment [1] Cell Types: MCF-7, T47D, NCI-H460, HCT116 and other cancer cells Tested Concentrations: 0.001-10 µM Incubation Duration: 72 h Experimental Results: Inhibited cancer cell proliferation, PI3KCA mutant MCF-7 cells were the most sensitive , IC50 is 2.25 µM. Western Blot Analysis[1] Cell Types: MCF-7 Tested Concentrations: 0.001-10 µM Incubation Duration: 1, 4, 24 and 48 hrs (hours) Experimental Results: Inhibition of pAKT and pGSK3β at various concentrations and time points for up to 48 hrs (hours).
Animal Protocol	Animal/Disease Models: balb/c (Bagg ALBino) mouse (carrying A549 xenografts) [1] Doses: 10, 30, and 100 mg/kg Route of Administration: Oral; Single Dose Experimental Results: Dose-dependent increase in total plasma concentrations, resulting in 24 hrs (hrs (hours)) There was almost complete loss of measurable pAKTS473 in tumors at all time points. Tumor growth inhibition (TGI) was 23%, 31% and 41% at doses of 10, 30 and 100 mg/kg, respectively. Animal/Disease Models: balb/c (Bagg ALBino) mouse (carrying PIK3CA mutant MCF-7 xenografts) [1] Doses: 3 or 10 mg/kg Route of Administration: Oral; one time/day for 49 days Experimental Results: vs. tamoxifen When combined with fenfen (5 mg/kg; one time/day), significant tumor regression was seen at 3 mg/kg and 50% at 10 mg/kg, respectively.
References	[1]. Identification and development of a subtype-selective allosteric AKT inhibitor suitable for clinical development. Sci Rep. 2022 Sep 20;12(1):15715.
Additional Infomation	VAD-044 is an allosteric AKT inhibitor developed by Vaderis Therapeutics AG. It is being investigated for the treatment of Hereditary Hemorrhagic Telangiectasia.

Solubility Data

Solubility (In Vitro)

DMSO : ~50 mg/mL (~120.34 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.02 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 25.0 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: ≥ 2.5 mg/mL (6.02 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.4069 mL	12.0343 mL	24.0685 mL
	5 mM	0.4814 mL	2.4069 mL	4.8137 mL
	10 mM	0.2407 mL	1.2034 mL	2.4069 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.