JH-II-127 is an orally bioactive LRRK2 inhibitor (antagonist) with high potency, selectivity and brain penetration. Its IC50s against wild-type LRRK2 and LRRK2-G2019S and mutant LRRK2-A2016T are 6 and 6, respectively. 2 and 48 nM. JH-II-127 inhibits Ser935 phosphorylation in all tissues of mice like the brain. JH-II-127 may be utilized in the research/study of Parkinson's syndrome.

Physicochemical Properties

Molecular Formula	C19H21CLN6O3
Molecular Weight	416.87
Exact Mass	416.136
CAS #	1700693-08-8
PubChem CID	112499966
Appearance	Light yellow to yellow solid powder
Density	1.5±0.1 g/cm3
Boiling Point	581.4±60.0 °C at 760 mmHg
Flash Point	305.4±32.9 °C
Vapour Pressure	0.0±1.6 mmHg at 25°C
Index of Refraction	1.701
LogP	-0.68
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	5
Heavy Atom Count	29
Complexity	566
Defined Atom Stereocenter Count	0
InChi Key	HUEKBQXFNHWTQQ-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H21ClN6O3/c1-21-16-15-12(20)10-22-17(15)25-19(24-16)23-13-4-3-11(9-14(13)28-2)18(27)26-5-7-29-8-6-26/h3-4,9-10H,5-8H2,1-2H3,(H3,21,22,23,24,25)
Chemical Name	[4-[[5-chloro-4-(methylamino)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]amino]-3-methoxyphenyl]-morpholin-4-ylmethanone
Synonyms	JH-II127; JH-II 127; JH-II-127
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	In HEK293 cells, JH-II-127 (0.03, 0.1, 0.3, 1, 3 µM; 90 minutes) suppresses LRRK2 [1]. In mouse Swiss 3T3 cells, endogenously produced LRRK2 is inhibited by JH-II-127 (0.3, 1, 3 µM; 90 minutes) [1].
ln Vivo	JH-II-127 (100 mg/kg; i.p.; single dosage) produces nearly full dephosphorylation of LRRK2 Ser935 in all tissues, including brain [1]. 1.19 Pharmacokinetic characteristics of JH-II-127 in wild-type male C57BL/6 mice [1]. Matrix route Tmax (h) C0/Cmax (ng/mL) AUCLast (h·ng/mL) AUGINF (h·ng/mL) T1/2 (h) CL (mL/min/kg) Vss (L/kg) Plasma IV (2 mg/kg) - 1604.47 532.67 535.57 0.66 62.24 1.73 Plasma PO (10 mg/kg) 1 802.72 3094.58 3867.07 - - - Brain IV (2 mg/kg) - 1343.6 239.31 246.47 0.23 1 35. 24 1.7 Brain PO (10 mg/kg) 1 247.35 688.21 762.38 - - -
Cell Assay	Western Blot Analysis[1] Cell Types: HEK293 cells (expressing GFP-LRRK2, GFP-LRRK2[G2019S], GFP-LRRK2[G2019S + A2016T] and GFP-LRRK2[A2016T], respectively. Tested Concentrations: 0.03, 0.1, 0.3, 1, 3 µM Incubation Duration: 90 minutes Experimental Results: Induced dose-dependent inhibition of Ser910 and Ser935 phosphorylation in wild-type LRRK2 and LRRK2[G2019S] stably transfected into HEK293 cells. For wild-type LRRK2 and LRRK2[G2019S], at ca. Inhibits phosphorylation of Ser910 and Ser935 at 0.3 μM. Induces dephosphorylation of Ser910 and Ser935 at concentrations of 0.3-1 μM in drug-resistant LRRK2[A2016T + G2019S] and LRRK2[A2016T] mutants. Western Blot Analysis[1] Cell Types: Mouse Swiss 3T3 Cell Tested Concentrations: 0.03, 0.1, 0.3, 1, 3 µM Incubation Duration: 90 min Experimental Results: Induces similar dose-dependent Ser935 dephosphorylation of endogenous LRRK2.
Animal Protocol	Animal/Disease Models: wild-type male C57BL/6 mice [1]. Doses: 2 mg/kg (intravenous (iv) (iv)injection); 10 mg/kg (oral); 10, 30, 100 mg/kg (intraperitoneal (ip) injection) Route of Administration: intravenous (iv) (iv)and intraperitoneal (ip) injection; oral administration; single. Experimental Results: intraperitoneal (ip) injection of 100 mg/kg resulted in near-complete dephosphorylation of LRRK2 Ser935 in all tissues, including the brain, and at 30 mg/kg resulted in near-complete inhibition in all tissues, but not at 10 mg/kg. There was only partial inhibition in the brain at kg doses. Exhibits good oral bioavailability.
References	[1]. Discovery of a Pyrrolopyrimidine (JH-II-127), a Highly Potent, Selective, and Brain Penetrant LRRK2 Inhibitor. ACS Med Chem Lett. 2015 Apr 7;6(5):584-9.

Solubility Data

Solubility (In Vitro)

DMSO : ~110 mg/mL (~263.88 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.75 mg/mL (6.60 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 27.5 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: ≥ 2.75 mg/mL (6.60 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 27.5 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	2.3988 mL	11.9941 mL	23.9883 mL
	5 mM	0.4798 mL	2.3988 mL	4.7977 mL
	10 mM	0.2399 mL	1.1994 mL	2.3988 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.