The chronic effect of the neuropeptide SP was analyzed by pretreatment of rats with 5 mg/kg of the neurokinin receptor 1 (NK1-R) antagonist aprepitant (a generous gift from Merck & Co) i.p. 12 h and 1 h before OX-7 injection and every 24 h after antibody administration. NK1-R is the physiological binding site of SP. Control animals were administered with vehicle or aprepitant administration without concomitant OX-7 injection.
Aprepitant
Manufactured by Merck Group
Sourced in United States
Aprepitant is a laboratory product manufactured by Merck Group. It functions as an active pharmaceutical ingredient (API) that is used in the development and production of pharmaceutical formulations.
Automatically generated - may contain errors
Lab products found in correlation
Sodium chloride (nacl), by Avantor (1 mentions)
Phosphoric acid, by Avantor (1 mentions)
Sodium hydroxide (naoh), by Avantor (1 mentions)
Lipofundin mct lct 20, by B. Braun (1 mentions)
Pepsin, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Sodium dihydrogen phosphate dihydrate, by Merck Group (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
Eflornithine, by Merck Group (1 mentions)
Phenothiazine, by Merck Group (1 mentions)
Clemastine fumarate salt, by Merck Group (1 mentions)
Spironolactone, by Merck Group (1 mentions)
Flunarizine hydrochloride, by Merck Group (1 mentions)
Bufexamac, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Compound c, by Merck Group (1 mentions)
11 protocols using aprepitant
1
Chronic Effect of Neuropeptide SP
2
NK1 Receptor Blockade in Acute Experiments
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In acute experiments, NK1-R blockade (NK1 receptors are the physiological binding site of SP) was achieved with the potent and highly selective NK1-R antagonist RP67580 as mentioned above [49 (link)]. For ease of use, we administered for chronic NK1-R blockade 5 mg/kg of the NK1-R antagonist aprepitant (a generous gift from Merck & Co) i.p. 12 h and 1 h before OX-7 injection and every 24 h after antibody administration [23 (link)]. Control animals were administered with vehicle or aprepitant administration without concomitant OX-7 injection.
3
Regulation of NK1R-FL Signaling by miR-206
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To detect the character of miR-206 in the regulation of NK1R-FL-dependent intracellular signaling, we monitored changes in the concentration of cellular Ins(1, 4, 5)P3. Thus, we selected SK-BR-3 and MCF-10A cell lines that express NK1R-FL as target groups and the MDA-MB-231 cell line, which does not express NK1R-FL, as a control group. These cells were treated with 10−7 M SP at different times. In some cases, cells transfected with miR-206 or As-miR-206 were incubated with the NK1R antagonist Aprepitant (C23H21F7N4O3, Merck) and then stimulated with 10−7 M SP. In other cases, the cells that had been transfected with miR-206 or As-miR-206 and subsequently stimulated with 10−7 M SP at the same time points. Then, the 3 groups of cells that with different treatments were all harvested and used to detect the Ins(1, 4, 5)P3 content with the d -myo-inositol-1,4,5-trisphosphate[3H] assay kit (GE healthcare, General Electric Co., USA). A standard curve was drawn according to the levels of Ins(1, 4, 5)P3.
4
Dissolution Method for Aprepitant
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Aprepitant was obtained as the European Pharmacopeia reference standard (code: Y0001825).
Acetonitrile and water of HPLC-grade were from Merck KGaA (Darmstadt, Germany). Sodium dihydrogen phosphate dihydrate of analytical grade was from Merck KGaA (Darmstadt, Germany). Phosphoric acid, sodium chloride and sodium hydroxide, also of analytical grade, were purchased from VWR chemicals (Leuven, Belgium). Pepsin was from Sigma-Aldrich (Lot # SLBQ2263V). EMEND® capsules were purchased from MSD SHARP & DOHME GMBH (Lot # MO 49340 and MO 45740 for the 80 mg and 125 mg, respectively, Haar, Germany). Lipofundin® MCT/LCT 20% was purchased from B. Braun (B. Braun Melsungen AG, Melsungen, Germany). FaSSGF/FaSSIF/FeSSIF, FeSSIF V2 and FaSSIF V3 powders were kindly donated by biorelevant.com (London, England).
Acetonitrile and water of HPLC-grade were from Merck KGaA (Darmstadt, Germany). Sodium dihydrogen phosphate dihydrate of analytical grade was from Merck KGaA (Darmstadt, Germany). Phosphoric acid, sodium chloride and sodium hydroxide, also of analytical grade, were purchased from VWR chemicals (Leuven, Belgium). Pepsin was from Sigma-Aldrich (Lot # SLBQ2263V). EMEND® capsules were purchased from MSD SHARP & DOHME GMBH (Lot # MO 49340 and MO 45740 for the 80 mg and 125 mg, respectively, Haar, Germany). Lipofundin® MCT/LCT 20% was purchased from B. Braun (B. Braun Melsungen AG, Melsungen, Germany). FaSSGF/FaSSIF/FeSSIF, FeSSIF V2 and FaSSIF V3 powders were kindly donated by biorelevant.com (London, England).
5
Aprepitant and SP Solubilization Protocol
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Aprepitant was purchased from Sigma–Aldrich (MO, USA) and dissolved in dimethyl sulfoxide (DMSO). SP was also purchased from Sigma–Aldrich Company and dissolved in distilled deionized water, containing 1% bovine serum albumin (BSA). The pH of the mixture was adjusted with 0.05 Molar acetic acid.
6
Glioblastoma Cancer Cell Culture Protocol
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U87 glioblastoma cancer cells were purchased from Pasteur Institute, Iran. Cells were maintained in RPMI-1640 medium (Gibco, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS) (Gibco, Grand Island, NY) and 1 mL of penicillin and streptomycin (Sigma 10,000 units penicillin and 10 mg of streptomycin/mL), incubated at 37°C with 5% CO2. SP and aprepitant were purchased from Sigma-Aldrich Company, St. Louis, MO, USA.
7
Esophageal Squamous Cell Carcinoma Cell Line Protocol
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The KYSE-30 human esophageal squamous cell carcinoma cell line was prepared from the National Cell Bank of Iran (NCBI), Pasteur Institute (Tehran, Iran). It was cultured in medium containing a 1 : 1 mixture of RPMI 1640, and Ham's F-12 (Betacell, Iran) supplemented with 5% fetal bovine serum and 1% penicillin-streptomycin (Grand Island, NY, USA) for 5-6 days until the cells reached the exponential phase of growth (0.6–1 × 106 cells/mL). SP and aprepitant were bought from Sigma-Aldrich (St. Louis, MO, USA). A stock solution of SP and aprepitant at the concentration of 70 μM and 74 mM, respectively, was prepared through dissolving the compound in sterile dimethyl sulfoxide (DMSO, Gibco, Scotland). It is then diluted to make a working solution and, cells were treated with relevant amounts of the SP and aprepitant solution to attain a concentration of different and specific doses. So, the final concentrations of DMSO in the culture medium did not exceed 0.1% in all the treatments.
8
Screening Cytotoxic Compounds in EP1/GFP Cells
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1 mL cultures of EP1/GFP reporter cells were plated at 100,000 cells/ml and treated with 33 μM of eflornithine (Sigma-Aldrich 1234249), phenothiazine (Sigma-Aldrich P14831), spironolactone (Sigma-Aldrich S3378), or vehicle control and counted daily by hemocytometer. Each day following counting, cells were diluted back to 100,000 cells/ml. For growth curve analysis by flow cytometry, 200 μl cultures of parasites were plated at 100,000 cells/ml and treated with 33 μM of phenothiazine (Sigma-Aldrich P14831), triprolidine hydrochloride (Sigma-Aldrich T6764), flunarizine hydrochloride (Sigma-Aldrich F8257), aprepitant (Sigma-Aldrich SML2215), bufexamac (Sigma-Aldrich B0760), clemastine fumarate salt (Sigma-Aldrich SML0445) or vehicle control. Parasites were stained daily with Sytox Orange and a fixed volume of cells was analyzed by flow cytometry. The number of live cells was calculated as the number of Sytox Orange negative cells falling within the live gate. Each day following flow analysis, cells were diluted back to 100,000 cells/ml according to the number of live cells counted.
9
Glioblastoma Cell Line Cultivation
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Experiments were performed using the U87 cell line, a human primary glioblastoma cell line. The cells were bought from the National Cell Bank of Institute Pasteur of Iran (Tehran, Iran). Cells were cultured in RPMI 1640 and Ham's F12 (RPMI/F12) media (Gibco-BRL, Life technology, Paisley, Scotland), supplemented with 10% fetal bovine serum (Gibco-BRL, Life technology, Paisley, Scotland) and 1% penicillin-streptomycin (Gibco-BRL, Life technology, Paisley, Scotland). SP and aprepitant were purchased from Sigma-Aldrich Company (St. Louis, MO, USA).
10
AMPK Inhibitor Effect on DRG-BMSC Co-culture
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The AMPK inhibitor, compound C, was purchased from Millipore (Merck, Billerica, MA, USA), and the dose of compound C was 20 μΜ [19 (link), 20 (link)]. compound C was added into the system for 24 hours after coculture for 8 days. The NK1 receptor antagonist, aprepitant, was purchased from Sigma-Aldrich (SML2215-5MG, Shanghai, China). The dose of aprepitant was 50 μΜ [21 (link), 22 (link)]. After coculture for 8 days, DRG and BMSCs were treated with aprepitant for 48 hours. Then, BMSCs were collected for the following analysis.
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