GHSR/Growth hormone secretagogue receptor

GH释放的脉动性明显受到下丘脑两种激素——GH释放激素（GHRH）和生长抑素的交替顺序变化的调节。这种搏动的夹带似乎涉及GH介导的负反馈。最近克隆了一种参与GH释放的新受体。GH释放肽和MK-0677激活该受体会启动和放大GH的搏动，并与含GHRH的弓状神经元中Fos免疫反应性和电活动的增加有关。我们发现，用GH预处理小鼠可以阻断MK-0677对这些神经元的激活。同样，奥曲肽抑制MK-0677的作用。为了确定这种GH介导的GHRH神经元负反馈是直接的，还是通过GH刺激脑室周围神经元释放生长抑素，我们选择性地灭活了五种特定生长抑素受体亚型（亚型2）之一的基因[2]。

循环GH水平的升高导致狗血清胰岛素样生长因子-1（IGF-1）水平的剂量相关增加。然而，尚不清楚全身IGF-1和GH水平的升高是否会影响这些激素的脑脊液（CSF）水平。因此，在狗身上设计了一项研究，以确定循环GH水平升高是否是GH促分泌素（MK-0677）的结果，或者外源性GH给药是否导致狗CSF中IGF-1和GH水平升高。根据体重，共有12只正常的年轻成年雄性狗被随机分为三个治疗组（每组4只狗）。有4只车辆控制犬。一组4只狗口服溶于去离子水中的MK-0677（5mg/kg/天）。第三组4只狗接受了猪生长激素（pGH）的皮下注射，剂量为0.1IU/kg/天。在开始治疗之前以及治疗的第7天和第15天，从所有狗身上收集血液和脑脊液样本。所有样本均使用经过验证的放射免疫测定法进行分析。施用MK-0677或pGH导致体重增加和血清IGF-1和GH水平升高，具有统计学意义（P<0.05）。相比之下，在研究的第7天或第15天，服用MK-0677不会导致CSF IGF-1或GH水平显著增加（P>0.05）。CSF IGF-1值范围为1.2至2.0 ng/ml，在研究过程中从每只狗身上采集的三个单独样本之间变化最小。同样，无论治疗组如何，所有狗的CSF GH水平都很低（<0.08 ng/ml至2.4 ng/ml）。这项研究表明，IGF-1或GH的循环水平与正常狗CSF中这些激素的水平之间没有相关性。载体对照犬血清和脑脊液IGF-1水平之间约100倍的差异表明，循环IGF-1存在血脑屏障。同样，尽管血清GH水平升高，但CSF GH水平没有升高，这表明正常狗的GH存在血脑屏障。这些结果表明，狗脑脊液中GH和IGF-1的可能来源是中枢神经系统[2]。

Compounds used were: MK-0677 (50 μg), octreotide (100 μg), and mGH (30 μg). Mice were give an initial ip injection (0.1 ml) of either saline, octreotide or mGH, followed 10 min later by an ip injection (0.1 ml) of either saline or MK-0677. Thus, the first study comprised of the following groups: saline/saline, saline/MK-0677, mGH/saline, mGH/MK-0677 saline/saline, saline/MK-0677, mGH/saline, mGH/MK-0677, and the second study of: saline/saline, saline/MK-0677, octreotide/saline, octreotide/MK-0677. Additionally, a number of mice were injected ip with hypertonic saline (0.2 ml, 1.5 M) to serve as positive controls for the immunocytochemistry. Ninety minutes after injection animals were terminally anesthetized with sodium pentobarbitone (60 mg/kg; ip) and perfused transcardially with heparinized saline followed by 4% paraformaldehyde in 0.1 M phosphate buffer (PB, pH 7.4). Brains were then removed and placed in the same solution for 24 h before being stored at− 80 C until processing. Coronal sections of forebrain (40 μm) were cut on a freezing microtome and placed in 0.1 M PB containing Triton X-100 (PB-T, pH 7.4). Sections were incubated for 24 h at 4 C in Ab-2 Fos antibody (rabbit polyclonal; 1:1000 in 1% normal sheep serum). The antibody-antigen complex was localized with a 1-h incubation in biotinylated anti-rabbit Ig (1:100), followed by a 1-h incubation in avidin, biotinylated horseradish peroxidase (1:50). The reaction product was visualized using a glucose oxidase-diaminobenzidine-nickel method, and Fos-like immunoreactivity was visualized as a dense purple-black precipitate restricted to the nucleus. The number of c-fos positive nuclei in the arcuate and periventricular nuclei (six to eight sections per mouse) were counted double-blind and a group mean calculated (mean ± SEM). Statistical analysis was performed by a two- way ANOVA followed by an all pairwise multiple comparison of data (Student-Newman-Keuls method) with significance taken as P < 0.05.[1]

[1]. Horm Metab Res. 1999 Feb-Mar;31(2-3):133-7.;
[2]. Mol Endocrinol. 1997 Oct;11(11):1709-17..

Ibutamoren is an orally bioavailable, small molecule, non-peptide growth hormone secretagogue (GHS). Upon administration, ibutamoren promotes the release of growth hormone (GH) from the pituitary gland, thereby increasing plasma GH levels. This may counteract GH deficiency. GH plays an important role in many biological processes.
See also: Ibutamoren Mesylate (has salt form).

C27H36N4O5S

528.66354

624.228

C, 61.34; H, 6.86; N, 10.60; O, 15.13; S, 6.06

159634-47-6

159752-10-0 (mesylate);159634-47-6;

178024

Typically exists as solid at room temperature

1.32g/cm3

1.631

3.94

130.42

2

7

8

37

920

1

CC(C(N[C@@H](C(N1CCC2(CN(S(=O)(C)=O)C3=CC=CC=C32)CC1)=O)COCC4=CC=CC=C4)=O)(N)C

UMUPQWIGCOZEOY-JOCHJYFZSA-N

InChI=1S/C27H36N4O5S/c1-26(2,28)25(33)29-22(18-36-17-20-9-5-4-6-10-20)24(32)30-15-13-27(14-16-30)19-31(37(3,34)35)23-12-8-7-11-21(23)27/h4-12,22H,13-19,28H2,1-3H3,(H,29,33)/t22-/m1/s1

(R)-2-amino-N-(3-(benzyloxy)-1-(1-(methylsulfonyl)spiro[indoline-3,4'-piperidin]-1'-yl)-1-oxopropan-2-yl)-2-methylpropanamide

L 163191; L-163191; Ibutamoren; 159634-47-6; Ibutamoren [INN]; UNII-GJ0EGN38UL; GJ0EGN38UL; 2-amino-2-methyl-N-[(2R)-1-(1-methylsulfonylspiro[2H-indole-3,4'-piperidine]-1'-yl)-1-oxo-3-phenylmethoxypropan-2-yl]propanamide; L 163191; CHEMBL13817; L163191; MK-677; MK 677; MK677

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

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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。