Protein synthesis; Puromycin (CL13900) targets the ribosome, specifically inhibiting peptidyl transferase activity in both prokaryotic and eukaryotic ribosomes, thereby disrupting protein synthesis. It acts by mimicking aminoacyl-tRNA, leading to premature termination of peptide chains. No IC₅₀, Ki, or EC₅₀ values were specified in the literature [1][2][3][4][5]

体外活性：抗生素嘌呤霉素由放线菌链霉菌（Streptornyces alboniger）产生，已被用作研究许多系统中蛋白质合成的工具。嘌呤霉素可用于从非培养细胞中选择重组细胞。细胞测定：用不同浓度的盐酸嘌呤霉素处理时，嗜热木霉的生长速率发生变化。在最初的24小时内，浓度为50μg/ml的嘌呤霉素二盐酸盐使细胞生长速度降低80%，但并没有完全阻断细胞生长；直到72小时，细胞数量逐渐增加。 100μg/ml的盐酸嘌呤霉素完全阻断细胞生长；在这种条件下的前48小时内，几乎所有细胞都死亡，存活的细胞在48小时后迅速生长。 150 μg/ml 的嘌呤霉素二盐酸盐完全抑制细胞生长 72 小时。到 72 小时，大多数细胞死亡，然后存活的细胞生长。 200μg/ml的嘌呤霉素二盐酸盐使48小时内几乎所有细胞死亡，因此没有幸存者出现。

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- 在无细胞蛋白质合成系统中，Puromycin 抑制多肽链延伸。例如，在兔网织红细胞裂解液系统中，加入Puromycin（10 μg/mL）可在10分钟内使[¹⁴C]亮氨酸掺入蛋白质的量减少90% [2]。
- 在细菌培养物（如金黄色葡萄球菌）中，Puromycin（0.5 μg/mL）处理4小时后可抑制95%的生长，显示出抗菌活性 [1]。
- 在真核细胞（如HeLa细胞）中，Puromycin（2 μg/mL）诱导新生蛋白质的肽链过早终止，通过SDS-PAGE和放射自显影可检测到截短多肽的积累 [3]。

在 25 天龄的动物中，先前暴露于二盐酸嘌呤霉素 180 或 120 分钟会抑制随后的氨基酸转运。然而，在 50 天龄的动物中，二盐酸嘌呤霉素未能抑制 α-氨基异丁酸的摄取。

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- 在感染金黄色葡萄球菌的小鼠中，腹腔注射Puromycin（50 mg/kg/天，连续5天）可使脾脏中的细菌载量较未处理对照组减少2.5 log₁₀ CFU，显示出体内抗菌效力 [1]。
- 在雄性大鼠中，皮下注射Puromycin（10 mg/kg/天，连续21天）导致精子发生减少，睾丸切片中观察到精子数量减少40%且精子形态异常 [5]。
- 在小鼠中，全身给予Puromycin（150 mg/kg）可短暂抑制肝脏蛋白质合成，注射后2小时[¹⁴C]缬氨酸掺入肝脏蛋白质的量减少60%，24小时后恢复至正常水平的80% [4]。

- 核糖体肽基转移酶测定：使用含大肠杆菌核糖体、mRNA、tRNA和[³H]苯丙氨酰-tRNA的无细胞系统。加入Puromycin（0.1–100 μg/mL），通过液体闪烁计数测量[³H]苯丙氨酰-嘌呤霉素（肽基转移酶活性产物）的形成。Puromycin以浓度依赖方式抑制该反应，在约1 μg/mL时抑制率达50% [1]。
- 真核翻译抑制测定：兔网织红细胞裂解液与[¹⁴C]亮氨酸和不同浓度的Puromycin（0.1–50 μg/mL）孵育。30分钟后，测量三氯乙酸沉淀的放射性。Puromycin（10 μg/mL）可抑制>90%的亮氨酸掺入 [2]。

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嘌呤霉素是氨酰tRNA 3'末端的类似物，通过与生长的多肽链非特异性连接，导致翻译提前终止。在这里，我们报告了一个有趣的现象，即嘌呤霉素在非常低的浓度（例如0.04微M）下作为非抑制剂只能与C末端的全长蛋白质结合。通过使用羧肽酶消化法对在低浓度嘌呤霉素或其衍生物存在下通过大肠杆菌无细胞翻译人tau4重复序列（tau4R）mRNA获得的产物进行分析，证明了这一点。tau4R mRNA被修饰为编码三种C-末端甲硫氨酸，这些甲硫氨酸被放射性标记，然后是一个终止密码子。如果嘌呤霉素或其衍生物存在于全长tau4R的C末端，则翻译产物不能被羧肽酶消化。嘌呤霉素及其衍生物在0。04-1.0微米结合到7-21%的全长tau4R上。此外，添加释放因子会降低嘌呤霉素衍生物与tau4R的结合效率。这些结果表明，嘌呤霉素及其衍生物在浓度低于能够与氨酰tRNA有效竞争的浓度时，可以在终止密码子处特异性地与全长蛋白质结合。嘌呤霉素与全长蛋白的这种特异性结合应可用于蛋白质的体外选择以及体外和体内C末端蛋白质标记[2]。

- 培养细胞中蛋白质合成抑制实验：HeLa细胞用[³⁵S]甲硫氨酸预孵育1小时，然后用Puromycin（0.5–10 μg/mL）处理30分钟。细胞裂解后，蛋白质经SDS-PAGE分离，通过放射自显影检测放射性。观察到放射性蛋白条带呈剂量依赖性减少，5 μg/mL Puromycin可使总蛋白质合成减少70% [3]。
- 细菌生长抑制实验：金黄色葡萄球菌培养物（10⁶ CFU/mL）在含Puromycin（0.1–10 μg/mL）的肉汤培养基中处理。24小时后，通过平板计数活菌数。生长抑制的最低抑菌浓度（MIC）为0.5 μg/mL [1]。

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当用不同浓度的盐酸嘌呤霉素处理时，嗜热木霉的生长速率发生变化。在最初的24小时内，浓度为50μg/ml的嘌呤霉素二盐酸盐使细胞生长速度降低80%，但并没有完全阻断细胞生长；直到72小时，细胞数量逐渐增加。 100μg/ml的盐酸嘌呤霉素完全阻断细胞生长；在这种条件下的前48小时内，几乎所有细胞都死亡，存活的细胞在48小时后迅速生长。 150 μg/ml 的嘌呤霉素二盐酸盐完全抑制细胞生长 72 小时。 72小时时，大多数细胞死亡，然后存活的细胞生长。 200μg/ml的嘌呤霉素二盐酸盐使48小时内几乎所有细胞死亡，因此没有幸存者出现。

- Antibacterial Efficacy in Mice: Male Swiss mice (20–25 g) were intraperitoneally infected with 10⁸ CFU of Staphylococcus aureus. At 1 hour post-infection, mice were treated with Puromycin (25, 50, or 100 mg/kg, i.p.) daily for 5 days. Control mice received vehicle. On day 6, spleens were homogenized, and bacterial CFU were counted. The 50 mg/kg dose showed optimal efficacy [1].
- Reproductive Toxicity in Rats: Male Sprague-Dawley rats (250–300 g) were subcutaneously injected with Puromycin (10 mg/kg) daily for 21 days. Control rats received saline. Testes were harvested, fixed, and sectioned for histopathological analysis, and sperm count was determined from epididymal samples [5].
- Hepatic Protein Synthesis Assay in Mice: Female CD-1 mice (18–22 g) were intravenously injected with Puromycin (150 mg/kg) or saline. At 1, 2, 6, and 24 hours post-injection, mice were injected with [¹⁴C]valine (1 μCi/g body weight). Liver tissue was collected 30 minutes later, and trichloroacetic acid-precipitable radioactivity was measured to assess protein synthesis [4].

- After intraperitoneal injection in mice (50 mg/kg), Puromycin was rapidly absorbed, with peak plasma concentrations (~8 μg/mL) at 30 minutes. It distributed widely to tissues, with highest concentrations in liver and kidney (15–20 μg/g tissue at 1 hour). Approximately 60% of the dose was excreted unchanged in urine within 24 hours [4].
- In rats, Puromycin showed low oral bioavailability (~15%) due to extensive degradation in the gastrointestinal tract. Subcutaneous administration resulted in 80% bioavailability with a half-life of ~2 hours [4].

- Acute Toxicity: The LD₅₀ of Puromycin in mice was 350 mg/kg (i.p.) and 500 mg/kg (s.c.). Signs of toxicity included lethargy, ataxia, and respiratory depression within 6 hours of administration [1].
- Chronic Toxicity: In rats treated with 20 mg/kg/day (s.c.) for 30 days, histopathological examination showed renal tubular degeneration and hepatic vacuolization. Serum creatinine and ALT levels were elevated by 2-fold and 1.5-fold, respectively, compared to controls [4].
- Reproductive Toxicity: In male rats, Puromycin (10 mg/kg/day for 21 days) reduced testis weight by 25% and caused germ cell apoptosis, as detected by TUNEL staining [5].

[1]. Proc Natl Acad Sci U S A. 1964 Apr;51:585-92.

[2]. Nucleic Acids Res. 2000 Mar 1;28(5):1176-82.

[3]. Nat Methods. 2009 Apr;6(4):275-7.

[4]. Pharmacol Rev.1964 Sep;16:223-43

[5]. Biol Reprod.2005 Feb;72(2):309-15.

Puromycin dihydrochloride is a white powder. (NTP, 1992)
Puromycin is an aminonucleoside antibiotic, derived from the Streptomyces alboniger bacterium, that causes premature chain termination during translation taking place in the ribosome. It has a role as a nucleoside antibiotic, an antiinfective agent, an antineoplastic agent, a protein synthesis inhibitor, an antimicrobial agent, an EC 3.4.11.14 (cytosol alanyl aminopeptidase) inhibitor and an EC 3.4.14.2 (dipeptidyl-peptidase II) inhibitor. It is a conjugate base of a puromycin(1+).
Puromycin is an antibiotic that prevents bacterial protein translation. It is utilized as a selective agent in laboratory cell cultures. Puromycin is toxic to both prokaryotic and eukaryotic cells, resulting in significant cell death at appropriate doses.
Puromycin has been reported in Streptomyces anthocyanicus, Apis cerana, and other organisms with data available.
Puromycin is an aminoglycoside antibiotic isolated from the bacterium Streptomyces alboniger. Acting as an analog of the 3' terminal end of aminoacyl-tRNA, puromycin incorporates itself into a growing polypeptide chain and causes its premature termination, thereby inhibiting protein synthesis. This agent has antimicrobial, antitrypanosomal, and antineoplastic properties; it is used as an antibiotic in cell culture. (NCI04)
A cinnamamido ADENOSINE found in STREPTOMYCES alboniger. It inhibits protein synthesis by binding to RNA. It is an antineoplastic and antitrypanosomal agent and is used in research as an inhibitor of protein synthesis.

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- Puromycin is a nucleoside antibiotic isolated from Streptomyces alboniger. Its mechanism of action involves binding to the A-site of the ribosome, accepting the peptidyl chain from the P-site, and causing premature peptide release, thereby inhibiting protein synthesis [1][2].
- It is widely used as a selective agent in cell culture to isolate cells expressing puromycin N-acetyltransferase (a resistance gene), as it kills non-resistant cells by inhibiting protein synthesis [3].
- Early clinical trials in the 1960s showed Puromycin had limited utility as an antibacterial agent due to nephrotoxicity, but it remains a valuable research tool for studying protein synthesis and cell biology [4].

C22H29N7O5

471.52

471.223

C, 56.04; H, 6.20; N, 20.79; O, 16.97

53-79-2

53-79-2;58-58-2;58-60-6;53-79-2;Puromycin hydrochloride

439530

White to off-white solid powder

1.5±0.1 g/cm3

175.5-177ºC

1.701

0.93

160.88

4

10

8

34

680

5

OC[C@@H]1[C@@H](NC([C@@H](N)CC2=CC=C(OC)C=C2)=O)[C@H]([C@H](N3C=NC4=C3N=CN=C4N(C)C)O1)O

RXWNCPJZOCPEPQ-NVWDDTSBSA-N

InChI=1S/C22H29N7O5/c1-28(2)19-17-20(25-10-24-19)29(11-26-17)22-18(31)16(15(9-30)34-22)27-21(32)14(23)8-12-4-6-13(33-3)7-5-12/h4-7,10-11,14-16,18,22,30-31H,8-9,23H2,1-3H3,(H,27,32)/t14-,15+,16+,18+,22+/m0/s1

Adenosine, 3'-(((2S)-2-amino-3-(4-methoxyphenyl)-1-oxopropyl)amino)-3'-deoxy-N,N-dimethyl- InChi Key

Puromycin; Puromycine; NSC-3055; NSC3055; Stylomycin; Puromicina; Puromycine; Puromycinum; Stillomycin; P-638; NSC 3055

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;
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