| 包装 | 价格(元) |
| 10mM (in 1mL DMSO) | 电议 |
| 10mg | 电议 |
| 50mg | 电议 |
Kinase experiment: | [35S]GTPγS binding assays are performed. CHO-K1 cells stably expressing the human H3 receptor (~400 fmol/mg protein) are homogenized in ice-cold buffer (50 mM Tris/HCl, pH 7.4). Homogenates are centrifuged twice (20,000g for 10 min at 4℃), and the final pellet is resuspended in 50 volumes of buffer. Membranes (550 μg of protein) are pretreated with adenosine deaminase (1 U/mL) and incubated for 60 min at 25℃ with 0.1 nM [35S]GTPγS and the drugs to be tested in a final volume of 1 mL of assay buffer (50 mM Tris/HCl, 50 mM NaCl, 5 mM MgCl2, 10 μM GDP, and 0.02% bovine serum albumin, pH 7.4). The nonspecific binding is determined using 10 μM nonradioactive GTPγS. Incubations are stopped by rapid filtration under vacuum through GF/B glass fiber filters. After washing with ice-cold water, the radioactivity trapped on filters is counted by liquid scintillation spectrometry. A similar assay is used to assess competitive antagonism. In brief, membranes (10 μg of protein) of HEK-293 cells stably expressing the human H3 receptor (~600 fmol/mg protein) are preincubated in presence of Pitolisant in the buffer (50 mM Tris/HCl, pH 7.4, 10 mM MgCl2, 100 mM NaCl, and 10 μM GDP) in a 96-well microplate under gentle agitation at room temperature (19-20℃) for 30 min before the addition of 0.1 nM [35S]GTPγS (final volume 200 μL). The nonspecific binding is determined using a 10 μM concentration of nonradioactive GTPγS. After 30 min, incubations performed in triplicate are stopped by rapid filtration under vacuum on a Multiscreen MAFCOB50 microplate. Radioactivity trapped on filters is counted by liquid scintillation spectrometry[1]. |
Animal experiment: | Mice[2] Adult female Albino Swiss mice weighing 20-22 g are used in the study. LY170053 or Pitolisant are suspended in 1 % Tween 80. The compounds or vehicle are administered intraperitoneally (i.p.) 30 min prior to the acute experiment. In the Pitolisant+LY170053 group, Pitolisant is administered 15 min before LY170053. Subchronic treatment is done at about 9:00 am (0.2 mL Tween to control group, Pitolisant-10 mg/kg b.w. to Pitolisant group, LY170053-2 mg/kg b.w. to LY170053 group, Pitolisant-10 mg/kg b.w. and LY170053 after 15 min-2 mg/kg b.w. to Pitolisant+LY170053 group) and at about 1:00 pm (LY170053 group and Pitolisant+LY170053 group).Rats[3] Male Wistar rats (220-300 g) receive vehicle (methylcellulose 1%, p.o.), and Pitolisant (10 mg/kg, p.o.). Ninety minutes later, they are killed by decapitation and nucleus accumbens are dissected out, weighed, frozen in liquid nitrogen and stored at -80℃. Tissues are homogenized in 1 mL of a 0.4 N perchloric acid/2.7 mM EDTA solution. After centrifugation (8000 rpm, 20 min, 4℃), supernatants are analysed by HPLC coupled to electrochemical detection. |
| 产品描述 | Pitolisant HCl (BF2.649; Ciproxidine) is a potent, novel, and selective nonimidazole inverse agonist at the recombinant human H3 receptor with a Ki value of 0.16 nM[1]. The histamine H3 receptor, a histamine autoreceptor, functions as a heteroreceptor that regulates the release of other neurotransmitters, making it an attractive drug target for a number of indications including cognition [2]. In vitro: BF2.649 behaved as a competitive antagonist with a Ki value of 0.16 nM. BF2.649 functioned as an inverse agonist with an EC50 value of 1.5 nM and an intrinsic activity about 50% higher than that of ciproxifan. Pitolisant in vitro potency was approximately 6 times lower at the rodent receptor [1]. In vivo: Pitolisant HCl was an oral bioavailable agonist. In mice, after oral and i.v. administrations of pitolisant HCl, the ratio of plasma areas under the curve was 84%. BF2.649 enhanced tele-methylhistamine levels in mouse brain, an index of histaminergic neuron activity in a dose dependent manner with an ED50 value of 1.6 mg/kg p.o. The response persisted after repeated administrations for 17 days [1]. Treatment with 20-, 40-, or 60-mg doses of pitolisant showed a statistically significant suppressive effect (standardized photosensitive response [SPR] reduction as measured with paired t-tests) in 9/14 (64%) patients of whom 6/14 (43%) showed abolition of the response to intermittent photic stimulation (IPS) [3]. BF2.649 showed significant inhibitory activity in several mouse models of schizophrenia [4]. References: |
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