AMG-3969 is a potent, novel and metabolically stable disruptor of glucokinase-glucokinase regulatory protein interaction (GK-GKRP) with IC50 of 4 nM. AMG-3969 exhibits potent cellular activity with an EC50 of 0.202 μM and IC50 of 4 nM. It potently reverses the inhibitory effect of GKRP on GK activity and promotes GK translocation in vitro (isolated hepatocytes). When administered to db/db mice, AMG-3969 demonstrated a robust pharmacodynamic response (GK translocation) as well as statistically significant dose-dependent reductions in fed blood glucose levels. Furthermore, with AMG-1694 and AMG-3969 (but not GK activators), blood glucose lowering was restricted to diabetic and not normoglycaemic animals. These findings exploit a new cellular mechanism for lowering blood glucose levels with reduced potential for hypoglycaemic risk in patients with type II diabetes mellitus.

Physicochemical Properties

Molecular Formula	C21H20F6N4O3S
Molecular Weight	522.46
Exact Mass	522.116
Elemental Analysis	C, 48.28; H, 3.86; F, 21.82; N, 10.72; O, 9.19; S, 6.14
CAS #	1361224-53-4
Related CAS #	1361224-53-4
PubChem CID	73053709
Appearance	White to off-white solid powder
Density	1.6±0.1 g/cm3
Boiling Point	648.8±65.0 °C at 760 mmHg
Flash Point	346.2±34.3 °C
Vapour Pressure	0.0±2.0 mmHg at 25°C
Index of Refraction	1.598
LogP	4.51
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	13
Rotatable Bond Count	4
Heavy Atom Count	35
Complexity	901
Defined Atom Stereocenter Count	1
SMILES	CC#C[C@@H]1N(C2=CC=C(C(C(F)(F)F)(O)C(F)(F)F)C=C2)CCN(S(=O)(C3=CN=C(N)C=C3)=O)C1
InChi Key	SIFKNECWLVONIH-INIZCTEOSA-N
InChi Code	InChI=1S/C21H20F6N4O3S/c1-2-3-16-13-30(35(33,34)17-8-9-18(28)29-12-17)10-11-31(16)15-6-4-14(5-7-15)19(32,20(22,23)24)21(25,26)27/h4-9,12,16,32H,10-11,13H2,1H3,(H2,28,29)/t16-/m0/s1
Chemical Name	2-[4-[(2S)-4-(6-aminopyridin-3-yl)sulfonyl-2-prop-1-ynylpiperazin-1-yl]phenyl]-1,1,1,3,3,3-hexafluoropropan-2-ol
Synonyms	AMG-3969; AMG 3969; AMG3969
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	Strong cellular activity is demonstrated by AMG-3969, which has an EC50 of 0.202 μM and an IC50 of 4 nM[1], [2]. In vitro (isolated hepatocytes), it effectively counteracts the inhibitory effect of GKRP on GK activity and stimulates GK translocation[3].
ln Vivo	According to research, AMG-3969 significantly lowers blood glucose levels in db/db mice in a dose-dependent manner and exhibits good in vivo pharmacokinetic (PK) properties in rats (75%). Blood glucose levels are significantly lowered by AMG-3969 (100 mg/kg), with a strong efficacy (56% reduction) seen at the 8-hour mark[2]. In three diabetes models—diet-induced obese (DIO), ob/ob, and db/db mice—AMG-3969 shows dose-dependent efficacy. In normoglycaemic C57BL/6 (B6) mice, however, AMG-3969 is ineffective in lowering blood glucose. When it comes to promoting carbohydrate substrate, AMG-3969 is very effective. After a single dosage, AMG-3969 shows prolonged changes to carbohydrate oxidation, as evidenced by an increased respiratory exchange ratio into the following day and night[3].
Animal Protocol	Formulated in 2% hydroxypropyl methycellulose, 1% Tween 80, pH 2.2 adjusted with MSA; 10, 30, 100 mg/kg; P.O. Diet induced obese (DIO), ob/ob and db/db mice
References	[1]. Antidiabetic effects of glucokinase regulatory protein small-molecule disruptors. Nature. 2013 Dec 19;504(7480):437-40. [2]. Small molecule disruptors of the glucokinase-glucokinase regulatory protein interaction: 3. Structure-activity relationships within the aryl carbinol region of the N-arylsulfonamido-N'-arylpiperazine series. J Med Chem. 2014 Apr 10;5. [3]. Small molecule disruptors of the glucokinase-glucokinase regulatory protein interaction: 2. Leveraging structure-based drug design to identify analogues with improved pharmacokinetic profiles. J Med Chem. 2014 Jan 23;57(2):325-38.

Solubility Data

Solubility (In Vitro)

DMSO:10 mM in DMSO Water:<1 mg/mL Ethanol:

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.79 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 (4.79 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 25.0 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (4.79 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	1.9140 mL	9.5701 mL	19.1402 mL
	5 mM	0.3828 mL	1.9140 mL	3.8280 mL
	10 mM	0.1914 mL	0.9570 mL	1.9140 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.