AH6809

EP/DP receptor

AH 6809（1 μM；30 分钟）抑制巨噬细胞中 T. serrulatus 毒液 (TsV) 诱导的和 PGE2 扩增的 IL-1β 和 cAMP 生成 [4]。 AH 6809 (30-300 μM) 拮抗全血中 PGD2 的抗聚集活性，表观 pA2 为 5.35[5]。

AH 6809（5 mg/kg；腹腔注射）可降低 TsV 诱导的死亡率、PGE2 和 IL-1β 的产生以及小鼠肺部中性粒细胞的浸润[4]。

1.在中国仓鼠卵巢细胞中稳定表达了八种类型和亚型的小鼠前列腺素受体，即前列腺素D（DP）受体、前列腺素F（FP）受体、前列环素I（IP）受体、血栓素A（TP）受体和前列腺素E受体的EP1、EP2、EP3和EP4亚型。通过测定放射性配体与相应受体结合的位移曲线中的Ki值，用32个前列腺素及其类似物检查了它们的配体结合特性。2.DP、IP和TP受体显示出很高的配体结合特异性，仅以高亲和力结合其自身假定的配体，如DP的PGD2、BW245C和BW868C，IP的蝉前列素、伊洛前列素和异卡巴环素，TP的S-145、I-BOP和GR 32191。3.FP受体结合PGF2α和氟前列醇，Ki值为3-4nM。此外，PGD2、17-苯基-PGE2、STA2、I-BOP、PGE2和M&B-28767以小于100nM的Ki值与该受体结合。4.EP1受体结合17-苯基-PGE2、舒前列酮和伊洛前列素以及PGE2和PGE1，Ki值为14-36nM。16,16-二甲基-PGE2和两种假定的EP1拮抗剂AH6809和SC-19220没有显示出与该受体的任何显著结合。M&B-28767，一种假定的EP3激动剂，以及米索前列醇，一种推定的EP2/EP3激动药，也以120 nM的Ki值与该受体结合。5.EP2和EP4受体显示出相似的结合特征。除了PGE2和PGE1外，它们还结合了16,16-二甲基PGE2和11-脱氧-PGE1。通过结合EP2受体但不结合EP4受体的丁前列素AH-13205和AH-6809，以及结合EP4但不结合EP 2受体的1-OH-PGE1来区分这两种受体。6.EP3受体显示出最广泛的结合谱，除了PGE2和PGE1外，还结合了舒前列酮、M&B-28767、GR63799X、11-脱氧-PGE1、16,16-二甲基-PGE2和17-苯基-PGE2，Ki值为0.6-3.7 nM。此外，三种IP配体伊洛前列素、卡巴环素和异卡巴环蛋白，以及一种TP配体STA2与该受体结合，Ki值分别与这些化合物对IP和TP受体的Ki值相当。7.8-Epi-PGF2α在10微M浓度下仅显示出与IP、TP、FP、EP2和EP3受体的弱结合[2]。

体外药物治疗
J774.1巨噬细胞以每孔2×105个细胞的密度接种在200μl添加了抗生素的无血清RPMI中。然后将细胞在37°C的5%CO2中培养2小时。接下来，去除上清液，用特定的抑制剂/拮抗剂处理或不处理细胞30分钟：吲哚美辛（10μM）；AH6809（1μM）；AH23848（1μM）；U-75302（0.1和1μM）；以及NFκB活化抑制剂（20 nM）。刺激前，在细胞培养基中加入H89二盐酸盐水合物（25μM）2小时。将乙醇储备溶液中的AH6809和U-75302稀释在细胞培养基中，仅向培养基中加入相同浓度的乙醇（最大0.1%）（对照）。将DMSO储备溶液中的AH23848和NFκB抑制剂稀释在细胞培养基中，仅向培养基中加入相同浓度的DMSO（最大0.1%）（对照）。所有化合物均在200μl无血清DMEM中稀释，并使用含有溶剂稀释剂的相同溶液作为对照。处理后，在相同的实验条件下用TsV（50μg ml−1）刺激细胞，在37°C、5%CO2的加湿气氛中24小时后，收集上清液进行IL-1β定量。[4]

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1.研究了AH6809（6-异丙氧基-9-氧杂蒽-2-羧酸）对各种药物对全血中人血小板的抗聚集和聚集作用的影响。2.前列腺素D2（PGD2）、BW245C、9α、11β-PGF2、PGI2和5'-N-乙基羧酰胺腺苷（NECA）均抑制ADP诱导的全血血小板聚集。AH6809拮抗PGD2、BW245C和9α、11β-PGF2的抗聚集活性，但不拮抗PGI2或NECA。AH6809拮抗NECA。3.AH6809对PGD2抗聚集活性的拮抗作用与浓度有关，可以通过增加PGD2的浓度来克服。数据分析得出AH6809的表观pA2为5.35。4.在比拮抗PGD2作用所需浓度高约10倍的浓度下，AH6809还拮抗了U-46619在全血中的聚集作用（pA2=4.45）。然而，浓度高达300微M的AH6809对ADP或血小板活化因子（Paf）诱导的聚集没有影响（pA2小于3.5）。5.在重新悬浮的血小板制剂中，AH6809对PGD2和U-46619的效力增加，表明该药物与血浆蛋白广泛结合。然而，在再悬浮的血小板中，AH6809相对于全血的特异性降低，因为ADP和Paf的聚集也被轻微拮抗。6.总之，AH6809似乎是一种弱但特异的人类血小板DP受体阻断药物，应被证明是一种有用的药物工具，可用于确定内源性PGD2在血小板聚集中的作用，并对抗聚集前列腺素的作用模式进行分类[5]。

IL-1r−/−, Casp1/11−/− and C57Bl/6 (WT) mice without treatment were inoculated with a sublethal or lethal dose of TsV (or PBS) as described above. Alox5−/− mice and 129sv mice were pre-treated or not with IL-1 receptor antagonist (IL-1Ra) at 10 mg kg−1, i.p., 1 h before and again 1 h after the sublethal or lethal TsV injection. IL-1Ra was kindly provided by Dr Stephen Poole, from the National Institute for Biological Standards and Control. In a specific experiment, the mice were either treated or not treated with MK886 (5-LO inhibitor, 5 mg kg−1 i.p., in 200 μl of 1% alcohol in water), indomethacin (COX1/2 inhibitor, 2 mg kg−1 i.p. in 200 μl of Tris[hydroxymethyl]aminomethane-HCl; TRIS-HCl, pH 8.2), SC-560 (selective COX1 inhibitor, 3 mg kg−1 i.p., in 200 μl of PBS), celecoxib (COX2 inhibitor, 5 mg kg−1 i.p., in 200 μl of water) or EP2 antagonist (AH6809, 5 mg kg−1 i.p., in 200 μl of PBS)67. The drugs (MK886 or indomethacin) or vehicles were administered four times, at 4 h and 0.5 h before and again 4 and 8 h after the lethal dose of TsV. The others drugs (SC-560, celecoxib, and EP2 antagonist) or vehicles were administered 1 day and again 1 h before the i.p. injection of lethal dose of TsV (180 μg kg−1). In other experiments, the Alox5−/− mice were treated or not with LTB4 (50 ng per mice, intranasal (i.n.) administration, in 20 μl of PBS). The LTB4 or vehicle (PBS+0.05% of ethanol) were administered 2 h and 0.5 h before the dose lethal of TsV (180 μg kg−1). The lungs were excised immediately after death or from mice survivors that were killed 8–12 h after the injection of TsV or vehicle. In some sets of experiments, two groups of mice were inoculated with PBS or a sublethal (120 μg kg−1) dose of TsV and, in only one, BAL fluids were collected 4 h later, to count the total cell number and neutrophils, as described previously47. In the other group, without BAL, the lungs were excised and weighed and 2 mg of tissue was homogenized in 2 ml of incomplete RPMI. After centrifugation (400g for 10 min), the supernatants were transferred to new tubes, split into two samples of 1 ml and stored at −80 °C until use. One sample was used for IL-1β and protein quantification analysis and the other for PGE2 and LTB4 measurement. For analysis of MPO activity, one lobule of a lung was cut out, immediately frozen in liquid nitrogen, and stored at −80 °C until use. In the therapeutic protocol, the lethal dose (180 μg kg−1) and superdose (360 μg kg−1) of TsV were injected and indomethacin (2 mg kg−1 i.p.) or vehicle were administered either 15 or 30 min after and again 4 and 8 h later. Mice survivors were killed 12 h after the envenomation.[4]

[1]. The utilization of recombinant prostanoid receptors to determine the affinities and selectivities of prostaglandins and related analogs. Biochim Biophys Acta. 2000 Jan 17;1483(2):285-93.

[2]. Ligand binding specificities of the eight types and subtypes of the mouse prostanoid receptors expressed in Chinese hamster ovary cells. Br J Pharmacol. 1997 Sep;122(2):217-24.

[3]. 6-Isopropoxy-9-oxoxanthene-2-carboxylic acid (AH 6809), a human EP2 receptor antagonist. Biochem Pharmacol. 1995 Nov 9;50(10):1731-3.

[4]. Opposing roles of LTB4 and PGE2 in regulating the inflammasome-dependent scorpion venom-induced mortality. Nat Commun. 2016 Feb 23;7:10760.

[5]. AH6809, a prostaglandin DP-receptor blocking drug on human platelets. Br J Pharmacol. 1988 Jul;94(3):745-54.

9-oxo-6-propan-2-yloxy-2-xanthenecarboxylic acid is a member of xanthones.

C17H14O5

298.29006

298.084

C, 68.45; H, 4.73; O, 26.82

33458-93-4

119461

White to off-white solid powder

1.3±0.1 g/cm3

514.2±50.0 °C at 760 mmHg

192.9±23.6 °C

0.0±1.4 mmHg at 25°C

1.618

3.91

76.74

1

5

3

22

446

0

O=C1C2=C(C=CC(C(O)=O)=C2)OC3=CC(OC(C)C)=CC=C31

AQFFXPQJLZFABJ-UHFFFAOYSA-N

InChI=1S/C17H14O5/c1-9(2)21-11-4-5-12-15(8-11)22-14-6-3-10(17(19)20)7-13(14)16(12)18/h3-9H,1-2H3,(H,19,20)

9-oxo-6-propan-2-yloxyxanthene-2-carboxylic acid

AH-6809; AH6809; AH 6809; 33458-93-4; AH 6809; 6-Isopropoxy-9-oxoxanthene-2-carboxylic acid; AH-6809; AH6809; 6-isopropoxy-9-oxo-9h-xanthene-2-carboxylic acid; 9-oxo-6-propan-2-yloxyxanthene-2-carboxylic acid; 6-(1-Methylethoxy)-9-oxo-9H-xanthene-2-carboxylic acid;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

DMSO: ~25 mg/mL (~83.8 mM)
H2O: ~0.1 mg/mL (~0.3 mM)

配方 1 中的溶解度: 2.5 mg/mL (8.38 mM) 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), 悬浮液；超声助溶。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到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: 2.5 mg/mL (8.38 mM) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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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。