Examorelin   中文名称: 艾沙瑞林

GHRF/growth hormone-releasing factor

Examorelin (Hexarelin)是一种合成的生长激素释放肽，具有心脏保护作用。众所周知，自噬的调节具有心脏保护作用，因此本研究探讨了自噬的作用和潜在的调节机制，以及六饥饿素在肥大心肌细胞中引发的抗心肌肥大作用。H9C2心肌细胞受到血管紧张素II（Ang II）的肥大。通过免疫荧光法测定自噬轻链-3（LC3）和细胞骨架蛋白。使用单核尸胺（MDC）进行自噬空泡可视化和Cyto ID染色进行自噬通量测量，也检测到自噬。通过Western印迹和qRT-PCR分析分子变化。采用流式细胞术和TUNEL法评估细胞凋亡。ATP含量和CCK-8测定用于评估细胞存活率的提高，而氧化应激则通过测量丙二醛（MDA）和超氧化物歧化酶（SOD）水平进行分析。Ang II诱导心肌细胞肥大、氧化应激、凋亡和细胞存活率降低，所有这些都被Examorelin（Hexarelin）治疗显著抑制，该治疗也增强了肥大H9C2细胞的自噬。此外，3-甲基腺嘌呤（3MA）对六角饥饿素诱导的自噬的抑制消除了六角饥饿肽的抗肥大功能，也消除了六对角饥饿素对细胞存活抑制和凋亡的保护作用。相反，自噬刺激剂雷帕霉素在H9C2肥大细胞中的应用抑制了细胞凋亡、细胞存活并减小了细胞大小。此外，hexarelin通过抑制哺乳动物雷帕霉素靶点（mTOR）的磷酸化来调节自噬的上游信号传导。我们提出，Examorelin (Hexarelin)通过与抑制mTOR信号通路相关的自噬依赖机制，在减轻心肌细胞肥大和凋亡方面发挥着新的作用。

急性心肌缺血再灌注损伤（IRI）是冠心病对心肌的不利影响。尽管再灌注疗法不断取得进展，但仍然缺乏有效的治疗策略来预防IRI。生长激素促分泌素（GHS）已被证明可以改善心血管疾病模型中的心脏功能、减轻炎症和调节自主神经系统（ANS）。最近，我们在心肌梗死的小鼠模型中证明了服用Examorelin（Hexarelin）/HEX后梗死面积减小。在本研究中，我们采用了再灌注缺血（IR）模型，以确定HEX是否会在具有更高临床相关性的模型中继续具有心脏保护作用。在C57BL/6 J小鼠中，通过短暂结扎左冠状动脉降支（tLAD）诱导心肌缺血，随后给予HEX（0.3 mg/kg/天；n=20）或赋形剂（VEH）（n=18）21天，首先在再灌注前立即给药。对IR损伤和假小鼠进行高场磁共振成像以评估左心室（LV）功能，与VEH治疗的小鼠相比，HEX治疗的小鼠LV功能有显著改善。21天后，HEX治疗的小鼠间质胶原、TGF-β1表达和肌成纤维细胞分化也显著降低。HEX治疗使ANS平衡向副交感神经为主转变；结合心肌肌钙蛋白I和TNF-α水平的显著降低，这些发现表明HEX对心肌具有抗炎作用。在这种IR模型中，HEX似乎可以重新平衡失调的ANS，激活迷走神经抗炎通路，以防止不良重塑和左心室功能障碍。针对IRI的干预措施有限，在改善急性心肌梗死（AMI）患者的临床结果方面取得了成功，这项研究为HEX的转化潜力提供了令人信服的证据，而其他所有干预措施在很大程度上都失败了[2]。

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Examorelin (Hexarelin)及其天然类似物ghrelin都具有相当的心脏保护活性。在实验性心肌梗死后的非常急性期给予单剂量的胃饥饿素对慢性心力衰竭的心功能有积极影响。因此，本研究旨在确定单剂量口服Examorelin（Hexarelin）在慢性疾病阶段是否具有相同的效果。雄性C57BL/6J小鼠通过左冠状动脉结扎产生心肌梗死或假手术，随后在手术后30分钟通过口服强饲法接受一剂hexarelin或赋形剂治疗。尽管两组心肌梗死后14天内的死亡率没有差异，但与赋形剂治疗相比，六饥饿素治疗在慢性期保护了心脏功能，表现为射血分数和缩短分数更高，肺重量/体重和肺重量/胫骨长度比更低。Hexarelin治疗同时降低了血浆肾上腺素和多巴胺水平，并将自主神经活动的平衡转向副交感神经活动，这可以从心率变异性分析中较小的低/高频功率比和较大的归一化高频功率中得到证明。结果首先表明，一剂口服六饥饿素治疗可能保护急性心肌梗死后的慢性心脏功能，并暗示激活生长激素促分泌素受体1a可能有利于心脏保护，尽管也可能涉及其他机制[3]。

细胞因子和心肌肌钙蛋白（CnT）-I测定[2]
在tLAD结扎或假手术后24小时和21天采集血液样本。血液被允许凝结，样本被离心。立即取出血清并储存在-80°C下，直至进行检测。24小时后测定血清CnT-I、白细胞介素-1β、IL-6和肿瘤坏死因子-α的浓度 根据制造商的说明，在术后h和21天使用MILLIPEX®图谱分析。

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HRV分析[2]
在tLAD结扎或假手术后21天进行HRV分析。使用生理分析系统记录心电图信号。用异氟烷麻醉小鼠，一旦心率（HR）稳定，记录心电图信号至少20分钟。通过HRV的功率谱分析检查ANS函数，其中HR用于使用快速傅里叶变换生成功率谱密度曲线。

Treatment administration [2]
Examorelin (Hexarelin)(0.3 mg/kg/day) or VEH was administered SC to each mouse immediately prior to reperfusion. Similarly, mice undergoing the sham procedure also received either VEH or Examorelin (Hexarelin) treatment. This dose was chosen based on previous studies demonstrating a cardioprotective effect. All mice then received their respective treatments once daily throughout the 21-day study period.

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Examorelin (Hexarelin) and vehicle administration [3]
Examorelin (Hexarelin) (600 μg per mouse) or vehicle was administered to 24 mice each by oral gavage 30 min after the MI procedure (hexarelin- or vehicle-treated group), and 10 mice after sham operation were also received oral vehicle administration (sham-operated group). The dose of oral hexarelin was chosen to be as effective as 40 μg/kg subcutaneous administration, which is approximately equimolar with that of ghrelin in the previous study.

[1]. Hexarelin protects cardiac H9C2 cells from angiotensin II-induced hypertrophy via the regulation of autophagy. Pharmazie. 2019 Aug 1;74(8):485-491.
[2]. Hexarelin targets neuroinflammatory pathways to preserve cardiac morphology and function in a mouse model of myocardial ischemia-reperfusion. Biomed Pharmacother. 2020 Jul:127:110165.
[3]. One dose of oral hexarelin protects chronic cardiac function after myocardial infarction. Peptides. 2014 Jun;56:156-62.

Conclusion: Modulating autonomic nervous system imbalances by enhancing parasympathetic tone has emerged as a promising therapeutic approach for ischemic heart disease and angina. The vagus nerve (VN) is considered to play a central role in the action of the growth hormone-releasing hormone agonist (GHS) and to act as an endogenous mechanism regulating immune responses and inflammation. In this study, we demonstrated that Examorelin (Hexarelin) treatment modulates the autonomic nervous system (ANS) and influences the ischemia-reperfusion (IR)-induced inflammatory response. Our results suggest that Examorelin (Hexarelin) may have significant implications for balancing the sympathetic and parasympathetic nervous systems and modulating adverse inflammatory pathways following acute myocardial infarction (AMI). Pharmacological stimulation of the VN with Examorelin (Hexarelin) may provide a novel cardioprotective strategy against ischemia-reperfusion injury (IRI) by inhibiting myofibroblast activation and left ventricular (LV) remodeling in AMI. [2]
Clinical Significance: Research in the field of cardioprotection has been plagued by numerous failures stemming from the failure to translate effective treatment strategies for preventing myocardial ischemia-reperfusion injury discovered in basic science laboratories into clinical applications. A major reason for this failure is the inappropriate use of experimental animal models. In our previous studies, we have clearly demonstrated the cardioprotective effect of Examorelin (Hexarelin) in a mouse model of permanent myocardial infarction (MI); however, the translational application prospects of this model are limited. In this study, using a clinically relevant model, Examorelin (Hexarelin) was administered before myocardial reperfusion (removal of coronary artery ligation sutures), and the results showed that Examorelin (Hexarelin) has good application prospects as an emerging drug for the prevention of myocardial ischemia-reperfusion injury (IRI). This protocol can be implemented in reperfusion centers. Therefore, this study makes an important contribution to the field of cardioprotection. In summary, although no improvement in overall mortality was observed, a single oral dose of Examorelin (Hexarelin) during the acute phase of myocardial infarction (MI) can still protect cardiac function during the chronic phase. In addition, it can reduce plasma adrenaline and dopamine levels and shift the balance of autonomic nervous activity towards parasympathetic activity. This study provides the first clear evidence to support oral Examorelin (Hexarelin) as a potential treatment for acute MI to protect cardiac function in the chronic phase. [3]

C47H58N12O6

887.040220000001

886.46

C, 63.64; H, 6.59; N, 18.95; O, 10.82

140703-51-1

6918297

H-His-D-Trp(2-Me)-Ala-Trp-D-Phe-Lys-NH2; L-histidyl-2-methyl-D-tryptophyl-L-alanyl-L-tryptophyl-D-phenylalanyl-L-lysinamide

HXAWFK

Typically exists as solid at room temperature

1.322 g/cm3

1403.6ºC at 760 mmHg

802.7ºC

0mmHg at 25°C

1.66

5.392

300.89

11

9

23

65

1600

6

CC1=C(C2=CC=CC=C2N1)C[C@@H](NC([C@@H](N)CC3=CN=CN3)=O)C(N[C@H](C(N[C@H](C(N[C@@H](C(N[C@H](C(N)=O)CCCCN)=O)CC4=CC=CC=C4)=O)CC5=CNC6=CC=CC=C56)=O)C)=O

RVWNMGKSNGWLOL-GIIHNPQRSA-N

InChI=1S/C47H58N12O6/c1-27-34(33-15-7-9-17-37(33)54-27)23-41(58-44(62)35(49)22-31-25-51-26-53-31)45(63)55-28(2)43(61)57-40(21-30-24-52-36-16-8-6-14-32(30)36)47(65)59-39(20-29-12-4-3-5-13-29)46(64)56-38(42(50)60)18-10-11-19-48/h3-9,12-17,24-26,28,35,38-41,52,54H,10-11,18-23,48-49H2,1-2H3,(H2,50,60)(H,51,53)(H,55,63)(H,56,64)(H,57,61)(H,58,62)(H,59,65)/t28-,35-,38-,39+,40-,41+/m0/s1

L-Lysinamide, L-histidyl-2-methyl-D-tryptophyl-L-alanyl-L-tryptophyl-D-phenylalanyl-

EP-23905; MF-6003; EP23905; Hexarelin; Examorelin; 140703-51-1; Examorelin [INN]; examorelina; examoreline; L-Lysinamide, L-histidyl-2-methyl-D-tryptophyl-L-alanyl-L-tryptophyl-D-phenylalanyl-; EP-23,905; MF6003; EP 23905; MF 6003

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。