Cycloheximide   中文名称: 放线菌酮

protein synthesis ( IC50 = 532.5 nM ); RNA synthesis ( IC50 = 2.88 μM )

体外活性：CHX诱导培养的外周血单核细胞凋亡，但对多形核细胞凋亡没有影响，因此其作用取决于细胞类型。对检测细胞凋亡的方法进行评价和比较，结果如下。吉姆萨染色的细胞离心涂片可以识别坏死和凋亡的主要形态特征。台盼蓝染色广泛用于估计细胞活力，但对于检测细胞凋亡毫无价值。两种荧光方法都提供了可靠且可重复的结果，并清楚地区分了凋亡细胞的亚群，并且密切相关。虽然提取 DNA 的琼脂电泳适用于多种细胞类型，但不能适用于所有细胞类型和凋亡条件。一般来说，吖啶橙/溴化乙锭染色细胞的显微镜检查可以被推荐为最可靠的测试方法。激酶测定：Cycloheximide (Naramycin A) 是一种真核生物蛋白质合成抑制剂，体内蛋白质合成和 RNA 合成的 IC50 值分别为 532.5 nM 和 2880 nM。细胞测定：通过将细胞与不同剂量的放线菌酮(CHX)一起孵育或通过312 nm UVB照射来诱导细胞凋亡。用于检测细胞凋亡的方法是光学显微镜（May Grunwald-Giemsa和台盼蓝染色）、荧光显微镜（吖啶橙/溴化乙锭和膜联蛋白V/碘化丙啶染色）和片段化基因组DNA的琼脂糖凝胶电泳。

在进行 200 µA 电击训练之前，小鼠接受 30、60 或 120 mg/kg 放线菌酮注射。放线菌酮对记忆测试试验中的潜伏期有显着影响（P<0.001）。在盐水对照中，这种电击水平导致测试试验的延迟明显高于训练时的延迟。注射最低测试剂量放线菌酮（30 mg/kg）导致测试试验的潜伏期显着高于盐水对照组。接受两种较高剂量放线菌酮中任一种的小鼠在试验中的潜伏期与盐水组相当，即在这些条件下较高剂量既不增强也不损害记忆，导致倒U型剂量反应曲线用于放线菌酮增强记忆。 HI 后 0 小时或 6 小时给予放线菌酮时，通过氯化三苯基四唑 (TTC) 对梗塞区域进行形态测量分析，梗塞体积显着减少 92% 和 61%，但如果给予放线菌酮，则梗死面积减少的趋势不显着。与 HI 对照组相比，在缺氧缺血 (HI) 后 12 小时施用，并且当延迟至 HI 后 24 小时施用时，没有观察到保护作用。

Cycloheximide（也称为 naramycin A）是一种真核蛋白质合成抑制剂，体内蛋白质合成和 RNA 合成的 IC50 值分别为 532.5 nM 和 2880 nM。

在 60 毫米培养皿上生长的 GF-1 细胞单层（4.0 mL，10⁵ 细胞/mL）培养至少 20 小时，然后用 PBS 冲洗两次。然后用放线菌酮 (CHX, 0.33 µg/mL) 和 BKA (20 µM) ANT 抑制剂处理细胞 0-5 天。最后，GSIV K1 菌株（感染复数 [moi] = 5）在初始步骤 (dpi) 后感染 0-5 天。孵育期结束后，将每个样品从培养基中取出并进行 PBS 清洗。之后，将细胞在 100 µL 染色溶液中孵育 10 至 15 分钟。

Mice: In this experiment, male ICR mice that are around two months old are utilized. IP cycloheximide is given in concentrations of 0, 30, 60, or 120 mg/kg (saline controls). Injections of cycloheximide are given half an hour before training. Mice amnesia is commonly studied at a dose of 120 mg/kg. Observe that rats receive much lower amnestic Cycloheximide doses (1-3 mg/kg) than mice do, which is consistent with a comparable difference in the LD50s of mice and rats. As measured 30–60 minutes after injection, cycloheximide doses of 120–150 mg/kg cause about 95% inhibition of brain protein synthesis, while doses of 30 mg/kg cause about 80% inhibition. Rats: Methoxyflurane anesthesia is used to perform unilateral carotid artery ligation on 7-day-old Sprague Dawley rat pups. The right common carotid artery is permanently ligated with 4-0 silk after a midline incision in the neck. Every animal received surgery for a maximum of five minutes. Rats are returned to their mother for recovery and feeding for two hours after surgery. The pups are then placed in an airtight chamber that is partially submerged in a temperature-controlled water bath to maintain a constant 36°C inside during the 100-minute hypoxic exposure period (8% O₂, 92% N₂). The rat pups in the HI with Cycloheximide treatment group receive an intraperitoneal injection of Cycloheximide at a dose of 0.6 mg/kg at0,6,12, or 24 hours of recovery, while the HI control group receives an equal volume of normal saline. Once the rat pups have been returned to their dam, they are sacrificed. The entire brain tissue is removed under deep intraperitoneal pentobarbital anesthesia (60 mg/kg) for triphenyl tetrazolium chloride (TTC) and flow cytometry at 48 and 72 hours after HI, respectively.

Interactions
CHLORDANE IS REPORTED TO CAUSE RAPID LOSS OF ACTIVITY /OF CYCLOHEXIMIDE/.
Pretreatment of rats with ... spironolactone & ethylestrenol ... protected against the fungicide cycloheximide.

---

Non-Human Toxicity Values
LD50 Rat oral 2 mg/kg
LD50 Rat ip 3700 ug/kg
LD50 Rat sc 2500 ug/kg
LD50 Rat iv 2 mg/kg
For more Non-Human Toxicity Values (Complete) data for CYCLOHEXIMIDE (10 total), please visit the HSDB record page.

[1]. Virus Res . 2016 Feb 2:213:37-45.

[2]. Behav Brain Res . 2012 Aug 1;233(2):293-7.

[3]. J Biol Chem . 1971 Jan 10;246(1):174-81.

[4]. Nucleic Acids Res. 2019 Jun 4;47(10):4974-4985.

Cycloheximide can cause developmental toxicity according to state or federal government labeling requirements.
Cycloheximide appears as colorless crystals. Used as a fungicide and as a anticancer drug. (EPA, 1998)
Cycloheximide is a dicarboximide that is 4-(2-hydroxyethyl)piperidine-2,6-dione in which one of the hydrogens attached to the carbon bearing the hydroxy group is replaced by a 3,5-dimethyl-2-oxocyclohexyl group. It is an antibiotic produced by the bacterium Streptomyces griseus. It has a role as a bacterial metabolite, a protein synthesis inhibitor, a neuroprotective agent, an anticoronaviral agent and a ferroptosis inhibitor. It is a member of piperidones, a piperidine antibiotic, an antibiotic fungicide, a dicarboximide, a secondary alcohol and a cyclic ketone. It is functionally related to a piperidine-2,6-dione.
Cycloheximide has been reported in Streptomyces, Streptomyces pulveraceus, and Streptomyces griseus with data available.
Antibiotic substance isolated from streptomycin-producing strains of Streptomyces griseus. It acts by inhibiting elongation during protein synthesis.

---

Mechanism of Action
STIMULATED TRANSPORT OF NUCLEAR RIBONUCLEOPROTEIN COMPLEXES TO CYTOPLASM AFTER ADMIN CYCLOHEXIMIDE TO RATS WAS STUDIED. ADMIN OF 2 MG/KG CYCLOHEXIMIDE CAUSED A DECR IN CONTENT OF TOTAL LIVER NUCLEAR RIBONUCLEOPROTEIN COMPLEX WITHIN 2 HR. THE OVERALL DECR WAS DUE TO AN INCREASED TRANSPORT INTO THE CYTOPLASM, NOT DECREASED SYNTHESIS. OTHER RESULTS SUGGEST THAT DURING INHIBITORY PHASE OF PROTEIN SYNTHESIS, GENE TRANSCRIPTION CONTINUES & GENE PRODUCT IS TRANSPORTED TO CYTOPLASM FOR TRANSLATION.
CYCLOHEXIMIDE (2-5 MG/KG BODY WT) CAUSED COMPLETE INHIBITION OF PROTEIN SYNTHESIS IN RAT LIVER WITHIN 30 MIN, & THE LABELING OF NUCLEAR PROTEINS WAS STRONGLY INHIBITED. UNDER THESE CONDITIONS, THE AMT OF NUCLEOLAR 45 S PRE-rRNA & ITS OROTATE-(14)C LABELING REMAINED UNAFFECTED FOR AT LEAST 4 HR, INDICATING THAT INITIALLY THE RATES OF SYNTHESIS & PROCESSING OF 45 S PRE-rRNA WERE NOT APPRECIABLY ALTERED. DRASTIC ALTERATIONS IN THE 45 S PRE-rRNA PROCESSING PATHWAYS OCCURRED AT THE EARLY STAGES OF CYCLOHEXIMIDE ACTION. THE CHANNELING OF NUCLEAR PRE-rRNA ALONG ALTERNATIVE PROCESSING PATHWAYS IS UNDER STRINGENT CONTROL BY THE CONTINUOUS SUPPLY OF CRITICAL PROTEINS.
CYCLOHEXIMIDE IS A POTENT INHIBITOR OF PROTEIN SYNTHESIS IN FUNGI & ANIMALS. IT CAUSES AN INCREASE IN ADRENAL RNA, INCREASED PRODUCTION OF GLUCOCORTICOIDS ... & DECREASE IN PYRUVATE UTILIZATION IN ISOLATED ADIPOSE TISSUE.

---

Therapeutic Uses
Antibiotics, Antifungal; Protein Synthesis Inhibitors
MEDICATION (VET): ANTIBIOTIC, ANTIFUNGAL, TRICHOMONACIDAL ... ESPECIALLY EFFECTIVE AGAINST MALIGNANT LYMPHOMAS IN DOGS.
Cycloheximide ... was used in the treatment of cryptococcal infections (Cryptococcus neoformans) before the development of amphotericin B.

C15H23NO4

281.35

281.162

C, 64.04; H, 8.24; N, 4.98; O, 22.75

66-81-9

6197

White to off-white solid powder

1.1±0.1 g/cm3

491.8±10.0 °C at 760 mmHg

111-116 °C

251.2±19.0 °C

0.0±2.8 mmHg at 25°C

1.499

Streptomyces

0.56

83.47

2

4

3

20

404

4

O=C(CC(C[C@H]([C@H]1C([C@@H](C)C[C@H](C)C1)=O)O)C2)NC2=O

YPHMISFOHDHNIV-FSZOTQKASA-N

InChI=1S/C15H23NO4/c1-8-3-9(2)15(20)11(4-8)12(17)5-10-6-13(18)16-14(19)7-10/h8-12,17H,3-7H2,1-2H3,(H,16,18,19)/t8-,9-,11-,12+/m0/s1

4-[(2R)-2-[(1S,3S,5S)-3,5-dimethyl-2-oxocyclohexyl]-2-hydroxyethyl]piperidine-2,6-dione

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 本产品在运输和储存过程中需避光。

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

配方 1 中的溶解度: ≥ 2.08 mg/mL (7.39 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 20.8 mg/mL澄清DMSO储备液加入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。

---

配方 2 中的溶解度: ≥ 2.08 mg/mL (7.39 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 20.8 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。

---

View More

配方 3 中的溶解度: ≥ 2.08 mg/mL (7.39 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 20.8 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

---

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。