VU10010 is a specific allosteric enhancer of M4 mAChR with EC50 of 400 nM. VU10010 binds to the allosteric site of M4 mAChR, increasing its affinity for acetylcholine and enhancing coupling to G protein. VU10010 increases carbachol-induced depression of excitatory transmission but not hippocampal synaptic transmission.

Physicochemical Properties

Molecular Formula	C17H16CLN3OS
Molecular Weight	345.85
Exact Mass	345.07
CAS #	633283-39-3
PubChem CID	714286
Appearance	White to off-white solid powder
LogP	5.05
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	3
Heavy Atom Count	23
Complexity	430
Defined Atom Stereocenter Count	0
InChi Key	FPRULFHDSFKYBV-UHFFFAOYSA-N
InChi Code	InChI=1S/C17H16ClN3OS/c1-9-7-10(2)21-17-13(9)14(19)15(23-17)16(22)20-8-11-3-5-12(18)6-4-11/h3-7H,8,19H2,1-2H3,(H,20,22)
Chemical Name	3-amino-N-[(4-chlorophenyl)methyl]-4,6-dimethylthieno[2,3-b]pyridine-2-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

ln Vitro

When compared to other mAChR subtypes (rM1, rM2, rM3, or rM5), VU10010 exhibits little activity but increases the M4 response to acetylcholine by a factor of 47. Increased affinity for ACh and M4-mediated [35S]GTPγS binding is brought about by VU10010[1]. At P2Y1R or mGluR5, VU10010 (10 μM) exhibits no agonist, antagonist, or allosteric potentiator activity[1]. When VU10010 is present, the EC50 values for ACh are 0.7 nM and 33 nM, respectively[1].

References

[1]. An allosteric potentiator of M4 mAChR modulates hippocampal synaptic transmission. Nat Chem Biol. 2008 Jan;4(1):42-50.

Solubility Data

Solubility (In Vitro)

DMSO: 125 mg/mL (361.43 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (6.01 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 20.8 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: ≥ 2.08 mg/mL (6.01 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 20.8 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	2.8914 mL	14.4571 mL	28.9143 mL
	5 mM	0.5783 mL	2.8914 mL	5.7829 mL
	10 mM	0.2891 mL	1.4457 mL	2.8914 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.