Rapamycin 雷帕霉素
| MDL | MFCD00867594 |
| EC | EINECS 262-640-9 |
| 别名 | 西罗莫司; RPM; Sirolimus |
| CAS | 53123-88-9 |
| 分子式 | C51H79NO13 |
| 分子量 | 914.18 |
| 纯度 | HPLC≥98% |
| 单位 | 瓶 |
| 生物活性 | Rapamycin (Sirolimus) 是一种特异性的 mTOR 抑制剂，IC50 为 0.1 nM。[1-5] |
| In Vitro | 雷帕霉素抑制HEK293细胞内源性mTOR活性，IC50为0.1 nM，比iRap和AP21967更有效，IC50分别为5 nM和10 nM [1]。雷帕霉素通过诱导自噬发挥其对恶性神经胶质瘤细胞的抗肿瘤作用，并且表明在恶性神经胶质瘤细胞中PI3K/Akt信号传导途径的破坏可以极大地增强mTOR抑制剂的有效性。雷帕霉素以剂量依赖性方式抑制所有三种细胞系中的细胞活力，但它们的敏感性不同。 T98G，U87-MG和U373-MG细胞的IC50水平分别为2 nM，1μM和>25μM[3]。 |
| In Vivo | 雷帕霉素（ip，1.5 mg/kg）治疗几乎完全可以防止7天和14天跖肌重量和纤维大小的肥大增加[4]。每天用WT或LS/+小鼠雷帕霉素（2mg/kg体重ip）处理4周，然后每周注射相同剂量另外4周。来自雷帕霉素处理的LS/+小鼠的心脏的异常胎儿基因表达谱显著逆转[5]。 |
| 激酶实验 | 将HEK293细胞以每孔2-2.5×10 5个细胞接种于12孔板中，仅在DMEM中血清饥饿24小时。将细胞模拟处理或用雷帕霉素（0.05-50nM），iRap（0.5-500nM）或AP21967（0.5-500nM）在37℃处理15分钟。在37℃下将血清加入终浓度为20％的30分钟。裂解细胞并通过SDS-PAGE分离细胞裂解物。将分离的蛋白质转移至PVDF膜，并用针对p70 S6激酶的Thr389的磷酸特异性一抗进行免疫印迹。使用ImageQuant和KaleidaGraph [1]分析数据。 |
| SMILES | O=C([C@@]1(O)[C@@H](CC[C@@H](C[C@@H](/C(C)=C/C=C/C=C/[C@H](C[C@@H](C)C([C@@H]([C@@H](/C(C)=C/[C@H]2C)O)OC)=O)C)OC)O1)C)C(N3CCCC[C@H]3C(O[C@@H](CC2=O)[C@@H](C[C@H]4C[C@H]([C@H](O)CC4)OC)C)=O)=O |
| 靶点 | mTOR |
| 动物实验 | 将雌性Sprague-Dawley大鼠（250-275g），成年雌性SD大鼠（225-250g）随机分配至治疗组或媒介物组，使得每组的平均起始体重相等。药物治疗开始于手术当天或14天停药后重新加载的第一天。雷帕霉素每天一次通过腹膜内注射以1.5mg/kg的剂量递送，溶于2％羧甲基纤维素中。 CsA每天一次通过皮下注射以15mg/kg的剂量递送，溶于10％甲醇和橄榄油中。 FK506每天一次通过皮下注射以3mg/kg的剂量递送，溶于10％乙醇，10％的cremophor和盐水中。将小鼠[5]雷帕霉素以20 mg/mL的浓度溶解于乙醇中，过滤灭菌，重悬于载体（0.25％PEG，0.25％吐温-80）中，浓度为1 mg/mL，腹腔注射（2 mg）/kg体重），每天4周或每天4周，然后每周注射另外4周。注射开始于8周（肥大发作前）或12周（表明确定肥大后）的年龄，并在治疗4周后或治疗8周后评估小鼠，详见结果和图例。 。作为对照，WT和LS/+小鼠仅用载体处理。 |
| 细胞实验 | HL-60，NB4，U937，KG-1和K562细胞常规传代于RPMI-1640中，补充有10％热灭活的FBS，2 mM L-谷氨酰胺，50 U/mL青霉素和50μg/ mL链霉素。在37°C下，5％CO2加湿的气氛。对于实验，通过离心收获指数生长的细胞，并重悬于含有10％FBS的新鲜培养基中。在各种浓度的DMSO或1μMATRA存在下，在BD Falcon六孔板中以2×10 5/mL的初始细胞密度接种细胞。在分化剂之前20分钟加入雷帕霉素（20nM）。在第2天，向每个孔中加入0.3mL新鲜培养基。通过台盼蓝排除法测定活细胞，并使用血细胞计数器进行定量[2]。 |
| 数据来源文献 | [1]. Edwards SR, et al. The rapamycin-binding domain of the protein kinase mammalian target of rapamycin is a destabilizing domain. J Biol Chem, 2007, 282(18), 13395-13401.
[2]. Lalic H, et al. Rapamycin enhances dimethyl sulfoxide-mediated growth arrest in human myelogenous leukemia cells. Leuk Lymphoma. 2012 Nov;53(11):2253-61. [3]. Takeuchi H, et al. Synergistic augmentation of rapamycin-induced autophagy in malignant glioma cells by phosphatidylinositol 3-kinase/protein kinase B inhibitors. Cancer Res, 2005, 65(8), 3336-3346. [4]. Bodine SC, et al. Akt/mTOR pathway is a crucial regulator of skeletal muscle hypertrophy and can prevent muscle atrophy in vivo. Nat Cell Biol, 2001, 3(11), 1014-1019. [5]. Marin TM, et al. Rapamycin reverses hypertrophic cardiomyopathy in a mouse model of LEOPARD syndrome-associatedPTPN11 mutation. J Clin Invest. 2011 Mar;121(3):1026-43. |
| 规格 | 10mg 10mM*1mL (in DMSO) 20mg |
Rapamycin是一种大环内酯类免疫抑制剂，通过不同的细胞因子受体阻断信号传导，阻断T淋巴细胞及其他细胞由G1期至S期的进程，从而发挥免疫抑制效应。还是一种有效且特异性的mTOR 抑制剂和自噬激活剂。
使用本产品的应用案例（仅供参考）：
In Vitro：
细胞实验：Wound Healing Assay： Cells at the logarithmic growth stage were placed in a six-well plate with a cell density of 1 × 105 cells/well and routine-cultured in an incubator at 37 °C with 95% humidity and 5% CO2 for 24 h. A 200-μL pipetting tip was used to vertically scratch the six-well plate to avoid tilting. The damaged or dead cells were washed away with PBS and then the 2 mL culture medium containing different concentrations of rapamycin was added to the treatment groups. The same amount of medium (culture medium and DMSO, the concentration of DMSO was the same as that of the 10−4 drug concentration group) was added to the negative control group. Photographs were taken at 0 and 24 h using a microscope and the wound healing area was calculated using ImageJ. The experiment was repeated thrice.
来源文献：Liu Y, Zhang J, Long J, Qiu X, Wang T, Wang J. The Effects of Rapamycin on the Proliferation, Migration, and Apoptosis of Human Tracheal Fibroblasts (HTrF) and Human Tracheal Epithelial Cells (HTEpiC). J Clin Med. 2022 Jan 25;11(3):608. doi: 10.3390/jcm11030608.
In Vivo：
种鸽，翼静脉注射
Experimental Design：A total of 90 pairs of breeding pigeons were randomly assigned to 3 groups. Each pair of breeders fed 4 young squabs (“2+4” feeing pattern). On the first day of the lactation period, the breeding pigeons were randomly subjected to one of the following treatments for 7 days: sham treatment (RAPA vehicle at an equivalent volume, control), low-dose RAPA (Jinpan,Beijing, China; 0.6 mg/kg body weight (BW), dissolved in absolute ethanol and diluted with 0.85% physiological saline, once daily at 09:00 hours, wing vein injection) and high-dose RAPA (1.2 mg/kg BW).
来源文献：Chen MJ, Fu Z, Jiang SG, Wang XQ, Yan HC, Gao CQ. Targeted disruption of TORC1 retards young squab growth by inhibiting the synthesis of crop milk protein in breeding pigeon (Columba livia). Poult Sci. 2020 Jan;99(1):416-422. doi: 10.3382/ps/pez513. Epub 2019 Dec 30. PMID: 32416826; PMCID: PMC7587900.
小鼠，腹腔注射
Mice were transfected with AdIGFBPrP1 into liver tissue via the tail vein; the recombinant adenoviral vector encoding IGFBPrP1 was synthesized. Concurrently, LY294002 (4 mg/kg) and rapamycin (2 mg/kg, Beijing Jinpan) were intraperitoneally injected for one month. Both doses were determined by pre-experimental results. The mice were sacrificed at 1, 2, 4, 8, and 12 weeks (n = 8), and liver tissues were harvested.
来源文献：Huang TJ, Ren JJ, Zhang QQ, Kong YY, Zhang HY, Guo XH, Fan HQ, Liu LX. IGFBPrP1 accelerates autophagy and activation of hepatic stellate cells via mutual regulation between H19 and PI3K/AKT/mTOR pathway. Biomed Pharmacother. 2019 Aug;116:109034. doi: 10.1016/j.biopha.2019.109034. Epub 2019 May 29. PMID: 31152924.