| 名稱(chēng) | RS 504393 |
| 描述 | RS 504393 is a highly selective CCR2 chemokine receptor antagonist (IC50s: 89 nM and > 100 μM for human recombinant CCR2 and CCR1). |
| 細(xì)胞實(shí)驗(yàn) | Briefly, cytosolic calcium influx was measured in CCR2-CHL cells loaded with the fluorescent dye Fura-2-AM. Quantitation of signal intensity used the integrated signal intensity for 82 s after the addition of chemokine and thus has units of M·s. Antagonism by various compounds of calcium influx was measured using an approximate ED50 dose of MCP-1 (3 nM) and an approximate ED25 dose for MCP-3 (5 nM). Chemotaxis was measured over 1 h using THP-1–5X cells in a 96-well Boyden chamber apparatus. Cell migration through the polycarbonate filter was quantified by fluorescent staining using propidium iodide in 0.1% Triton X-100. These assays typically gave stimulated to unstimulated migration of 6-fold, range 4–10-fold, using a maximally effective concentration of MCP-1. Chemotaxis antagonist measurements used 3 nM MCP-1 or RANTES; these concentrations are near the ED95 attractant concentration for MCP-1 and for RANTES as agonists. The data are expressed by normalization to the uninhibited migration caused by the agonist chemokine. The antagonist was present in both chambers of the Boyden apparatus [1]. |
| 激酶實(shí)驗(yàn) | Briefly, binding was measured using membranes prepared from two cell lines, THP-1 and CCR2-CHL cells. Each competition assay was composed of cell membranes, 50 pM 125I-MCP, MCP buffer, protease inhibitors, and test compound. Equilibrium was achieved by incubation at 28?°C for 90 min. Membrane-bound 125I-MCP was collected by filtration through GF/B filters presoaked in polyethyleneimine and bovine serum albumin, followed by four rapid washes with approximately 0.5 ml of ice-cold buffer containing 0.5 M NaCl and 10 mM HEPES, pH 7.4. MCP buffer consists of 50 mMHEPES, pH 7.2, 1 mM CaCl2, 5 mMMgCl2, and 0.1% bovine serum albumin. Protease inhibitors include 0.1 mM phenylmethylsulfonyl fluoride, 1 μM leupeptin, and 0.35 mg/ml pepstatin. THP-1 cells are a human monocyte cell line that express both CCR1 and CCR2. CCR2-CHL cells are Chinese hamster lung cells that have been stably transformed with an expression vector bearing the human CCR2b receptor [1]. |
| 動(dòng)物實(shí)驗(yàn) | To evaluate the therapeutic effects of MCP-1/CCR2 signaling, either propagermanium (3 or 8 mg/kg orally once a day) or RS-504393 (2 mg/kg orally twice a day) was mandatorily injected into their mouths to wild-type mice from 3 days before ureteral ligation until the day of sacrifice. In addition, to determine the viability for the usage of CCR2 antagonists for the treatment of renal fibrosis, propagermanium (8 mg/kg) was given daily, beginning 4 days after ureter ligation. For pathological examination, both the obstructed and contralateral kidneys were harvested from UUO animals 4, 7, and 14 days after ureteral ligation (n = 5 at each time point). Untreated age-matched male wild-type mice and CCR2-deficient mice were used as normal control (n = 6 for each group). Since propagermanium treatment was started from 3 days before ureteral ligation, mice treated with propagermanium for 3 days at day 0 were used as a negative control (n = 5) [3]. |
| 體外活性 | RS-504393抑制了CCR2-CHL細(xì)胞中由MCP-1刺激的鈣流入，其IC50值為35 nM [1]。作為CCR2的拮抗劑，RS 504393的處理顯著抑制了過(guò)敏原誘導(dǎo)的β-己糖苷酶釋放。這種抑制效果可通過(guò)補(bǔ)充重組MCP-1蛋白（100 pg/mL）[2]部分逆轉(zhuǎn)。 |
| 體內(nèi)活性 | 相較于接受了載體處理的小鼠明顯展現(xiàn)出急性炎癥跡象，RS 504393處理的小鼠卻未表現(xiàn)出即時(shí)過(guò)敏的臨床跡象。在天真（naive）動(dòng)物中未觀察到任何效果。RS 504393處理的小鼠中肥大細(xì)胞的脫顆粒作用顯著受到抑制，但對(duì)天真小鼠無(wú)影響[2]。與CCR2缺陷小鼠所獲得的結(jié)果類(lèi)似，RS-504393的治療顯著減輕了腎臟病理變化，特別是通過(guò)減少I(mǎi)型膠原蛋白合成介導(dǎo)的廣泛間質(zhì)纖維化[3]。 |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 10 mg/mL (23.95 mM)
H2O : Insoluble
|
| 關(guān)鍵字 | CC chemokine receptor | RS 504393 | RS504393 | Inhibitor | inhibit | CCR | RS-504393 |
| 相關(guān)產(chǎn)品 | BMS-817399 | Pirfenidone | WAY-639418 | PF-4136309 | AZD2098 | Aplaviroc | Vercirnon | CCR2 antagonist 5 | MLN-3897 TFA | CCR3 antagonist 1 | INCB-9471 | CCR6 antagonist 1 |
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