Y5 receptor ( Ki = 1.3 nM ); Y2 receptor ( Ki = 200 nM ); Y1 receptor ( Ki > 4000 nM )

CGP71683 盐酸盐是一种竞争性神经肽 Y5 受体拮抗剂，Ki 为 1.3 nM，对细胞膜上的 Y1 受体（Ki，>4000 nM）和 Y2 受体（Ki，200 nM）无明显活性[1]。
目前，除了CGP 71683A外，还没有其他的Y5拮抗剂公开可用，因此在研究Y5受体的作用时，它被用作参考化合物。在细胞系和分离器官的功能测定中，CGP 71683A已被证明是Y5受体的竞争性拮抗剂(Criscione等人，1998；Duhault et al., 2000；Dumont et al., 2000)。在本研究中，它对豚鼠Y5受体具有高亲和力，但对Y1和Y2亚型没有明显的活性（表1）。[1]
据报道，CGP 71683A与5-羟色胺（5-HT）再摄取识别位点和胆碱能毒蕈碱受体结合，与与Y5受体结合的亲和力几乎相同（Della Zuana et al., 2001）。[1]

CGP71683（15 nmol/大鼠，静脉注射，每天两次）显示出厌食作用，减少进食大鼠的食物摄入量和体重。 CGP71683 导致禁食大鼠的血清总 T4 比禁食对照大鼠高 3 倍，游离 T4 增加 37%[2]。

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神经肽Y （NPY）在进食状态下抑制TRH神经元，空腹时下丘脑NPY的高表达被认为与空腹诱导的下丘脑-垂体-甲状腺（HPT）轴的抑制有关。我们研究了中枢Y5受体在控制促甲状腺素（TSH）和甲状腺激素（TH）分泌中的作用。喂食和禁食大鼠每天两次在第三脑室中央注射Y5受体拮抗剂(CGP71683；15nmol/大鼠)72h。禁食大鼠在禁食72h结束时接受CGP71683单次中央注射（15nmol/rat）。在喂食大鼠中，Y5受体阻断使总摄食量减少32%，体重减少近10% (p<0.01)，证实了该受体在食物摄入控制中的作用。空腹72h大鼠在单次注射Y5拮抗剂1h后血清TSH升高4倍（p<0.001）。在禁食72小时内多次注射，Y5阻断导致甲状腺轴活化，血清T4升高3倍（p<0.001）， TSH和T3不变。在喂养大鼠中，慢性中央给药CGP71683导致血清总T4降低，但游离T4和TSH没有变化。在喂食和禁食的大鼠中，血清瘦素和PYY未因NPY中枢阻断而改变，这表明这些激素在观察到的改变中没有作用。因此，中枢Y5神经传递的抑制导致禁食期间甲状腺轴的激活，这表明NPY-Y5受体参与了禁食诱导的TSH和TH抑制。[2]

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在本研究中，CGP 71683A对豚鼠Y5受体具有高亲和力，但对Y1和Y2亚型没有明显的活性（表1）。用CGP 71683A预处理豚鼠后，对NPY的摄食反应明显减弱，表明该化合物是通过Y5受体介导的NPY诱导摄食的有效拮抗剂。先前的啮齿动物数据也显示了CGP 71683A对瘦和肥胖动物npy诱导的食物摄入的抑制作用(Della Zuana等，2001；Duhault et al., 2000；Polidori et al., 2000)以及糖尿病、24小时禁食和自由喂养动物的自发食物摄入(Criscione et al., 1998；Kask et al., 2001)。[1]

结合试验[1]
利用转染了编码豚鼠受体的质粒的细胞系，研究了配体的选择性和亲和力。表1中所示的激动剂数据先前已被报道过(Berglund et al., 1999；Lundell et al., 2001；Sharma et al., 1998)。拮抗剂BIBO 3304、h409 /22和CGP 71683A在相同方案制备的膜组分上进行测试。为了进行结合实验，将解冻后的受体膜重悬在含有2.5 mM CaCl2、1 mM MgCl2和2 g l−1杆菌肽的25 mM HEPES缓冲液（pH 7.4）中，并使用Ultra-Turrax均质机均质。在终体积为100 μl的条件下，与2 - 10 μg蛋白和125I-PYY（猪）在室温下结合2 h。非特异性结合被定义为在100 nM未标记的NPY存在下孵育后仍与细胞匀浆结合的放射性量。在竞争性研究中，不同浓度的非肽化合物BIBO 3304、H 409/22或CGP 71683A与125I-PYY一起被纳入孵育混合物中。各实验均以肽NPY为参考。使用TOMTEC细胞收割机，将GF/C过滤器预先浸泡在0.3%聚乙烯亚胺中，通过快速过滤终止培养。滤网用5ml 50 mM Tris （pH 7.4）在4℃下洗涤，60℃下干燥。干燥的滤片用MeltiLex A熔化闪烁片处理，滤片上保留的放射性使用Wallac 1450 betatplate计数器计数。使用Prism软件包对结果进行分析。

Rats: CGP71683 is dissolved in 30% DMSO and, prior to the experiment, stored frozen at -20°C. Every microinjection comprises 2 microliters of either 30% DMSO vehicle or CGP71683 (7.5 nmol/μL; 15 nmol/rat) injected via the guide cannula over a 30- to 60-second period, adhering to the following protocols: I-Rats given free access to chow are given six microinjections (15 nmol/rat, spaced 10–14 hours apart) and are killed one hour after the final injection, which occurs between 9 and 10 a.m. The amount of food consumed is calculated by calculating the daily reduction in chow mass (g) right before each intravenous injection. II: After fasting for 72 hours, rats are given a single microinjection of either vehicle or CGP71683 (15 nmol/rat) and are then killed one hour later. III - Rats are given multiple injections of vehicle or CGP71683 over the course of a 72-hour fast, following the same protocol as fed animals. The fasting period began 10 hours prior to the first microinjection. Rats are decapitated at the conclusion of the experiments, and serum is extracted from the trunk blood to measure the hormone concentrations[2].

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BIBO 3304 (30 nmol per animal), H 409/22 (100 – 200 nmol per animal) or saline were infused i.c.v. in a volume of 5 – 7 μl given 15 min before NPY (3.6 nmol in 5 μl) or saline. When the effect of CGP 71683A (60 nmol per animal) was tested, 30% dimethylsulphoxide (DMSO) was used as the vehicle. CGP 71683A or DMSO were infused in a volume of 10 μl 15 min before NPY (3.6 nmol in 5 μl) or saline. Food consumption (4 h response) and eating parameters were measured as above. [1]

[1]. Receptor subtypes Y1 and Y5 mediate neuropeptide Y induced feeding in the guinea-pig. Br J Pharmacol. 2002 Apr;135(8):2029-37.

[2]. Central NPY-Y5 receptors activation plays a major role in fasting-induced pituitary-thyroid axis suppression in adult rat. Regul Pept. 2011 Nov 10;171(1-3):43-7.

Our observation that increased locomotion might be connected to Y5 receptor stimulation is in agreement with the previous finding showing that the Y5 antagonist, CGP 71683A, decreases exploratory behaviour in rats (Kask et al., 2001). The suppression of food intake after CGP 71683A administration is not, however, related to the changes in locomotor activity (Kask et al., 2001). It has been reported that CGP 71683A binds to the serotonin (5-HT) re-uptake recognition site and cholinergic muscarinic receptors with virtually identical affinity to its binding to Y5 receptors (Della Zuana et al., 2001). Furthermore, chronic administration of CGP 71683A may produce local inflammatory changes near the site of injection (Della Zuana et al., 2001). Due to these non-specific actions and its poor solubility, CGP 71683A is not a good tool to use in vivo, and new selective Y5 antagonists are needed to characterize the physiological importance of the Y5 receptor. Recent reports on novel agents blocking Y5 receptors do not entirely support the view that Y5 receptors are crucial for NPY-induced feeding in rats (Kanatani et al., 2000a; Polidori et al., 2000). Therefore, our results with CGP 71683A in the guinea-pig should be regarded with caution until investigated with other Y5 antagonists. [1]

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Interestingly, CGP71683-treated fasted rats lost more weight than vehicle-treated fasted rats, supporting an involvement of Y5 receptors in energy expenditure independent of food consumption. Other studies have proposed that NPY via Y5 receptors decreases energy expenditure by affecting thermogenesis in brown and white adipose tissue. [2]

C26H30CLN5O2S

512.07

511.181

C, 60.98; H, 5.91; Cl, 6.92; N, 13.68; O, 6.25; S, 6.26

192322-50-2

192321-23-6; 192322-50-2 (HCl)

9849276

White to off-white solid powder

747.2ºC at 760 mmHg

405.7ºC

1.19E-22mmHg at 25°C

7.489

118.38

4

7

7

35

751

0

Cl[H].S(C1=C([H])C([H])=C([H])C2=C([H])C([H])=C([H])C([H])=C12)(N([H])C([H])([H])C1([H])C([H])([H])C([H])([H])C([H])(C([H])([H])N([H])C2N=C(C3=C([H])C([H])=C([H])C([H])=C3N=2)N([H])[H])C([H])([H])C1([H])[H])(=O)=O

DIQDKUNCSVFGHH-UHFFFAOYSA-N

InChI=1S/C26H29N5O2S.ClH/c27-25-22-9-3-4-10-23(22)30-26(31-25)28-16-18-12-14-19(15-13-18)17-29-34(32,33)24-11-5-7-20-6-1-2-8-21(20)24;/h1-11,18-19,29H,12-17H2,(H3,27,28,30,31);1H

N-[[4-[[(4-aminoquinazolin-2-yl)amino]methyl]cyclohexyl]methyl]naphthalene-1-sulfonamide;hydrochloride

CGP-71683A HCl; CGP71683A; CGP-71683A; CGP 71683A

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，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: 100~160 mg/mL (195.3~312.5 mM)

配方 1 中的溶解度: ≥ 2.67 mg/mL (5.21 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 26.7 mg/mL的澄清DMSO储备液加入到400 μL PEG300中并混合均匀；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: ≥ 2.67 mg/mL (5.21 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 26.7 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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配方 3 中的溶解度: ≥ 2.67 mg/mL (5.21 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (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 26.7 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。