human recombinant DP2 ( Ki = 0.013 μM ); rat recombinant DP2 ( Ki = 0.003 μM ); human native DP2 ( Ki = 0.004 μM )

体外活性：OC000459 抑制 [3H]PGD2 与转染人 DP2 的 CHO 细胞膜的结合，Ki 为 13 nM。 OC000459 还可以替换人 Th2 淋巴细胞膜上的 [3H]PGD2，Ki 为 4 nM。 OC000459 在表达 DP2 的完整 CHO 细胞中以浓度依赖性方式拮抗 PGD2 介导的钙动员，IC50 为 28 nM。 OC000459 抑制人 Th2 细胞响应 PGD2 (10 nM) 的趋化性，IC50 为 28 nM。 OC000459 (< 3 μM) 在分离的白细胞制剂和肝素化人全血中竞争性拮抗 PGD2 的作用。 OC000459 抑制嗜酸性粒细胞形状变化对 DK-PGD2 的反应，IC50 为 11 nM。 OC000459 (1 μM) 抑制 Th2 细胞和嗜酸性粒细胞响应肥大细胞上清液的激活。细胞测定： Timipiprant (OC000459)（0.0001 μM-10 μM；5 小时）抑制人 Th2 淋巴细胞的趋化性 (IC50=0.028 μM) 和人 Th2 淋巴细胞的细胞因子产生 (IC50=0.019 μM)。 Timipiprant (OC000459) (1 μM) 抑制 Th2 细胞和嗜酸性粒细胞响应 IgE/抗 IgE 激活的人肥大细胞上清液的激活。

OC000459以2mg/kg口服剂量给予Sprague-Dawley大鼠，显示血浆半衰期为2.9小时，达到最大血浆浓度的时间为1.3小时，达到的最大血浆浓度为1.54μg/mL。在注射 DK-PGD2 前 0.5 小时口服 OC000459 导致大鼠血液嗜酸性粒细胞增多呈剂量依赖性减少，ED50 为 0.04 mg/kg。在注射 DK-PGD2 前 0.5 小时口服 OC000459 也会导致大鼠中嗜酸性粒细胞积累的剂量依赖性抑制 ED50 为 0.01 mg/kg。根据全面分析 (FA) 人群和按方案 (PP) 人群的分析，中度持续性哮喘患者服用 OC000459（200 毫克，每天两次，持续 28 天）显示生活质量得到改善。在这些患者中，OC000459 改善了夜间症状评分，减少了痰液嗜酸性粒细胞几何平均数和呼吸道感染。 OC000459（200 mg，每日两次）治疗可抑制未使用类固醇的哮喘患者后期的哮喘反应和过敏原后痰嗜酸性粒细胞的增加。

OC000459对[3H]PGD2与人DP2结合的影响。[1]
OC000459抑制了[3H]PGD2与转染有人DP2的CHO细胞膜的结合，Ki为0.013±0.002μM（n=13个独立实验），如图2A所示。OC000459还从人Th2淋巴细胞的膜上置换了[3H]PGD2（Ki=0.004±0.001μM；n=3个独立实验），表明该化合物对天然受体具有活性，如图2B所示。 OC000459对大鼠重组DP2有活性（0.003±0.001μM；n=5个独立实验），但没有。。。

Timipiprant (OC000459)（0.0001 μM-10 μM；5 小时）抑制人 Th2 淋巴细胞趋化性 (IC50=0.028 μM) 和细胞因子产生 (IC50=0.019 μM)。 timapiprant (OC000459) (1 μM) 可抑制 Th2 细胞和嗜酸性粒细胞响应 IgE/抗 IgE 激活的人肥大细胞上清液的激活。

This was a two-centre, double-blind, randomised, placebo-controlled, crossover study. Eligible subjects were randomised to receive OC000459 200 mg orally twice daily or matching placebo for 16 days, in addition to inhaled salbutamol as required. The washout period was 3 weeks between treatment periods. The measurement of vital signs (heart rate and blood pressure) and FEV1 during the treatment periods are shown in table 1; these measurements were performed pre-dose on days 1, 8, 15 and 16, while exhaled nitric oxide fraction (FeNO) was performed pre-dose on days 1, 8 and 15. Pre-dose blood sampling was performed for the measurements of biochemistry and haematology at days 1, 8 and 15, with pharmacokinetics also measured on days 8 and 15. Seven subjects provided blood samples for the measurement of eosinophil shape change up to 8 h post-dose on day 8. An inhaled allergen challenge was performed on day 15 at 3 h post-dose. On day 16, a methacholine challenge was performed at 3 h post-dose, followed by induced sputum; methacholine challenge and induced sputum were therefore performed 24 h post-allergen challenge. [2]

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Thirty-six healthy male subjects with a history of grass pollen allergic rhinoconjunctivitis were recruited for the study. All subjects were tested for skin prick positivity to grass pollen and levels of grass pollen–reactive IgE were measured by radioallergosorbent test. After giving informed consent, subjects were randomised to receive OC000459 then placebo or placebo then OC000459. Thirty-five subjects completed both treatment periods.

The study design is shown in Fig. 1. The study was a single centre, randomised, double-blind, placebo-controlled, two-way crossover trial conducted out of grass pollen season. Patients underwent screening at least 14 days prior to the first dosing to ensure they were eligible to take part in the study. There were two treatment periods, each 8 days in duration. Group A received OC000459 (200-mg bid, given as two 100-mg capsules) followed by matching placebo and Group B received placebo first followed by OC000459. Each treatment period was separated by a washout period of at least 7 days to ensure drug was no longer present at the start of the second period. In practice, the washout period was 19–23 days (20.9 ± 0.7 days, mean ± SD).[3]

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Dissolved in 10% DMSO/saline solution; 10 mg/kg; Oral administration

Sprague-Dawley rats

[1]. Pharmacologic profile of OC000459, a potent, selective, and orally active D prostanoid receptor 2 antagonist that inhibits mast cell-dependent activation of T helper 2 lymphocytes and eosinophils. J Pharmacol Exp Ther. 2012.

[2]. Inhibition of the asthmatic allergen challenge response by the CRTH2 antagonist OC000459. Eur Respir J. 2013 Jan;41(1):46-52.

[3]. The CRTH2 antagonist OC000459 reduces nasal and ocular symptoms in allergic subjects exposed to grass pollen, a randomised, placebo-controlled, double-blind trial. Allergy. 2012 Dec;67(12):1572-9.

OC000459 is under investigation for the treatment of Severe Eosinophilic Asthma. OC000459 has been investigated for the treatment of Bronchial Asthma.

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D prostanoid receptor 2 (DP₂) [also known as chemoattractant receptor-homologous molecule expressed on T helper 2 (Th2) cells (CRTH2)] is selectively expressed by Th2 lymphocytes, eosinophils, and basophils and mediates recruitment and activation of these cell types in response to prostaglandin D₂ (PGD₂). (5-Fluoro-2-methyl-3-quinolin-2-ylmethylindo-1-yl)-acetic acid (OC000459) is an indole-acetic acid derivative that potently displaces [³H]PGD₂ from human recombinant DP₂ (K(i) = 0.013 μM), rat recombinant DP₂ (K(i) = 0.003 μM), and human native DP₂ (Th2 cell membranes; K(i) = 0.004 μM) but does not interfere with the ligand binding properties or functional activities of other prostanoid receptors (prostaglandin E₁₋₄ receptors, D prostanoid receptor 1, thromboxane receptor, prostacyclin receptor, and prostaglandin F receptor). OC000459 inhibited chemotaxis (IC₅₀ = 0.028 μM) of human Th2 lymphocytes and cytokine production (IC₅₀ = 0.019 μM) by human Th2 lymphocytes. OC000459 competitively antagonized eosinophil shape change responses induced by PGD₂ in both isolated human leukocytes (pK(B) = 7.9) and human whole blood (pK(B) = 7.5) but did not inhibit responses to eotaxin, 5-oxo-eicosatetraenoic acid, or complement component C5a. OC000459 also inhibited the activation of Th2 cells and eosinophils in response to supernatants from IgE/anti-IgE-activated human mast cells. OC000459 had no significant inhibitory activity on a battery of 69 receptors and 19 enzymes including cyclooxygenase 1 (COX1) and COX2. OC000459 was found to be orally bioavailable in rats and effective in inhibiting blood eosinophilia induced by 13,14-dihydro-15-keto-PGD₂ (DK-PGD₂) in this species (ED₅₀ = 0.04 mg/kg p.o.) and airway eosinophilia in response to an aerosol of DK-PGD₂ in guinea pigs (ED₅₀ = 0.01 mg/kg p.o.). These data indicate that OC000459 is a potent, selective, and orally active DP₂ antagonist that retains activity in human whole blood and inhibits mast cell-dependent activation of both human Th2 lymphocytes and eosinophils.[1]

C21H17FN2NAO2

371.360059499741

370.11

C, 68.10; H, 4.35; F, 5.13; N, 7.56; Na, 6.21; O, 8.64

950688-14-9

Timapiprant; 851723-84-7

11462174

Light yellow to yellow solid powder

4.4

55.1

1

4

4

26

516

0

XKRNYIKRDAGPQZ-UHFFFAOYSA-M

InChI=1S/C21H17FN2O2.Na/c1-13-17(11-16-8-6-14-4-2-3-5-19(14)23-16)18-10-15(22)7-9-20(18)24(13)12-21(25)26;/h2-10H,11-12H2,1H3,(H,25,26);/q;+1/p-1

2-[5-fluoro-2-methyl-3-(quinolin-2-ylmethyl)indol-1-yl]acetate

OC000459 sodium; OC000459; Timapiprant sodium; 950688-14-9; Timapiprant sodium (OC000459); Timapiprant (sodium); 950688-14-9 (sodium); OC-000459; OC 000459; Timapiprant; Timapiprant sodium

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中（例如氮气保护），避免吸湿/受潮。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ~100 mg/mL (~270.0 mM)

配方 1 中的溶解度: ≥ 2.08 mg/mL (5.62 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 20.8 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.08 mg/mL (5.62 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。