SKF89976A_hydrochloride_DataSheet_MedChemExpress

www.MedChemExpress.com

Data Sheet

Product Name:Cat. No.:CAS No.:

Molecular Formula:Molecular Weight:Target:Pathway:Solubility:

SKF89976A (hydrochloride)HY-100228A85375-15-1C22H26ClNO2371.90

GABA Receptor; GABA Receptor

Neuronal Signaling; Membrane Transporter/Ion ChannelDMSO: ≥ 34 mg/mL

BIOLOGICAL ACTIVITY: 

SKF89976A hydrochloride is a selective GABA transporter (GAT–1) inhibitor with IC50s of 0.28 μM, 137.34 μM and 202.8 μM for GAT–1,GAT–2 and GAT–3 in CHO cells, respectively.

IC50 & Target: IC50: 0.28 μM (GAT–1), 137.34 μM (GAT–2), 202.8 μM (GAT–3)[1]

In Vitro: SKF89976A has a weak antiallodynic action. SKF89976A weakly inhibits serotonin transporter (SERT), noradrenaline

transporter (NET), and dopamine transporter (DAT) in chinese hamster ovary (CHO) cells stably expressing each transporter using asubstrate uptake assay, with IC50 values of 3514 , 202.13, and 728.8, respectively[1]. SKF89976A is a GABA–transport blocker. GABA(100 μM), but is unaffected by 100 μM picrotoxin. 100 μM SKF 89976–A is known to block the transport of GABA into cells,

(1 mM) elicited an inward current that is completely suppressed by the GABA transport inhibitors tiagabine (10 μM) and SKF89976Acompletely eliminated the GABA–elicited current in a reversible fashion[2]. SKF89976A is a nontransportable blockers of GAT–1.SKF89976–A also suppresses baseline inward currents that likely result from tonic GAT activation by background GABA. SKF89976A(100 μM) reversibly reduces GAT currents in every studied cell by 67.9±4.4% (n=19). Intracellular perfusion of 20 μM SKF89976–Aprogressively reduced and blocked GABA–induced GAT currents without blocking GABAAR–mediated currents (n=4)[3.dose–dependently ameliorates the reduction in the withdrawal threshold in PSL model mice[1].

In Vivo: SKF89976A produces a weak antiallodynic response when administered i.v. (0.3 mg/kg). The i.t. injection of SKF89976A

PROTOCOL (Extracted from published papers and Only for reference)

Cell Assay: SKF89976A is dissolved in DMSO and stored, and then diluted with appropriate medium before use[1]. [1]CHO cellsstably expressing the mouse GAT subtypes, rat serotonin transporter (SERT), rat noradrenaline transporter (NET), and rat dopaminetransporter (DAT) are incubated with 10 nM tritium–labeled GABA or monoamines for 10 min in the absence or presence of variousconcentrations of the GAT inhibitors (e.g., SKF89976A) tested. Values presented for SERT, NET, and DAT are the mean±S.E.M. for 3experiments, with each being performed in duplicate[1].DMSO is less than 0.5%)[1]. [1]Mice[1]

Animal Administration: SKF89976A is dissolved in DMSO and diluted appropriately with ACSF or saline (the final concentration of5–week–old ddY male mice, weighing 25–30 g at the beginning of the study are used. Mice are administered NNC05–2090,SKF89976A, (S)–SNAP5114, or amitriptyline. NNC05–2090, SKF89976A, and (S)–SNAP5114 are dissolved in DMSO and dilutedappropriately with ACSF or saline (the final concentration of DMSO is less than 0.5%). The composition of ACSF (in mM) is 142 mMNaCl, 5 mM KCl, 2 mM CaCl2, 2 mM MgCl2, 1.25 mM NaH2PO4, 10 mM d–glucose, 10 mM HEPES, and 0.05% fatty acid–free bovineserum albumin (pH 7.4). The intraperitoneal (i.p.) injection of drugs is administered in a volume of 0.1 mL/10 g body weight. Whengiven intravenously (i.v.), solutions are injected into the tail vein in a volume of 0.1 mL/10 g body weight. The head of a mouse isplaced into a plastic cap and the body is held with one hand for an intrathecal (i.t.) injection. A 27–gauge needle attached to aHamilton microsyringe is inserted into the subarachnoid space between the L5 and L6 vertebrae of the conscious mouse and 5 mL of

www.MedChemExpress.com

the drug solution is slowly injected.

References:

[1]. Jinzenji A, et al. Antiallodynic action of 1–(3–(9H–Carbazol–9–yl)–1–propyl)–4–(2–methyoxyphenyl)–4–piperidinol (NNC05–2090), a betaine/GABAtransporter inhibitor. J Pharmacol Sci. 2014;125(2):217–26.

[2]. Kreitzer MA, et al. Glutamate modulation of GABA transport in retinal horizontal cells of the skate. J Physiol. 2003 Feb 1;546(Pt 3):717–31.[3]. Barakat L, et al. GAT–1 and reversible GABA transport in Bergmann glia in slices. J Neurophysiol. 2002 Sep;88(3):1407–19.

Caution: Product has not been fully validated for medical applications. For research use only.

Tel: 609-228-6898     Fax: 609-228-5909     E-mail: tech@MedChemExpress.com

Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA

www.MedChemExpress.com