Phorbol 12-myristate 13-acetate (PMA)   中文名称: 伏波酯

PKC ( EC50 = 11.7 nM ); NF-κB; SphK; protein kinase C (PKC)

为了检查 PKC 在 p38MAPK 磷酸化中的作用，使用 PKC 激活剂 Phorbol 12-myristate 13-acetate (PMA) (100 nM) 刺激细胞，该激活剂模拟 PKC 的天然激活剂 DAG 与 PKC 的 C1 区域的结合。在两种细胞类型中观察到PMA对p38MAPK的磷酸化，类似在αT3-1细胞中观察到的GnRH，即缓慢持续激活（30分钟时分别为3.2倍和3.6倍）。PKCs的不同定位可以解释 GnRH 和 PMA 激活的 PKCs 对 p38MAPK 磷酸化中发挥不同作用。一般来说，αT3-1 细胞中 GnRH 和 PMA 刺激 p38MAPK 磷酸化是由电压门控 Ca2+ 通道和 Ca2+ 激发介导的，而在正极化的LβT2 促进性腺激素细胞中，它仅由 Ca2+ 激励诱导[2]。 THP-1 细胞通过 PMA (200 ng/mL；1-5 天) 存在下脓液形成巨噬细胞样细胞 (THP-1巨噬细胞），从而导致形态变化和细胞表面表达增加为特征的巨噬细胞样表型 CD11 和 CD14[3]。 在单核细胞系 THP-1 中，根据死亡、增殖消失、乳胶珠的吞噬作用，PMA 导致比 VD3 更分泌的表型，以及 CD11b 和 CD14 的表达[5]。

诱发耳部水肿模型描述（PMA诱导法）
1. 致病机制：
该模型通过佛波醇-12-肉豆蔻酸酯-13-乙酸酯（PMA）激活蛋白激酶C（PKC）信号通路，进而：
• 刺激磷脂酶A2（PLA2）活化
• 促进前列腺素等炎症介质释放
• 引起局部血管扩张和通透性增加
• 导致炎性细胞浸润和组织水肿

2. 标准造模方案：
实验动物：Swiss品系雌性小鼠
体重范围：25-30克
实验分组：需设立溶剂对照组
给药参数：
• 药物浓度：100 μg/mL PMA（溶于适当溶剂）
• 给药体积：20 μL
• 给药部位：单侧耳廓（左耳或右耳）
• 给药方式：局部均匀涂抹于耳廓内外表面
• 观察时程：通常给药后4-6小时达峰效应

3. 模型验证指标：
3.1 主要评判标准：
• 耳厚度差异：游标卡尺测量，造模耳较对照耳增厚≥50%
• 血管通透性：伊文思蓝渗出量显著增加（定量检测）
3.2 辅助评判指标（可选）：
• 组织学检查：炎性细胞浸润程度
• 炎性因子检测：TNF-α、IL-1β等促炎因子水平
• 髓过氧化物酶（MPO）活性测定

4. 注意事项：
• 需严格控制环境温度（22±2℃）和湿度
• 建议在固定时间段进行实验以减少昼夜节律影响
• 溶剂对照组应使用相同体积的PMA溶解溶剂
• 动物需提前适应环境至少3天
• 操作时避免机械刺激影响耳部状态

5. 模型特点：
• 造模成功率：＞90%
• 炎症峰值时间：给药后4-6小时
• 持续时间：24-48小时内可观察到明显炎症反应
• 适用研究：抗炎药物筛选、炎症机制研究等
注：具体实验参数可根据研究目的进行调整，但需在文献中明确说明修改依据。建议初次建立模型时进行预实验确定最佳条件。

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诱发足部水肿模型（PMA诱导法）研究方案
致病机制
PMA通过激活蛋白激酶C（PKC）信号通路，刺激磷脂酶A2（PLA2）活化，促进前列腺素等炎症介质释放，引起局部血管扩张和通透性增加，最终导致炎性细胞浸润和组织水肿。该机制与肾病综合征等病理状态下的水肿形成具有相似性。

实验动物选择
• 大鼠模型：Wistar品系雄性成年大鼠，体重200-220g
• 小鼠模型：Swiss白化病雄性小鼠，体重25-30g

造模方法
1. 给药参数：
o 药物浓度：2.5μg PMA溶于20μL溶剂
o 给药部位：单侧耳廓局部涂抹
o 给药方式：单剂量给药

2. 实验设计要点：
o 需设立溶剂对照组
o 其他抑制性产品应在处死前4小时处理

模型验证指标
主要评判标准
• 外观监测：左右耳质量差异显著增大（游标卡尺测量）
• 生化指标：刺激巨噬细胞产生超氧阴离子（定量检测）

辅助评判指标（可选）
• 血管通透性变化（伊文思蓝渗出法）
• 炎性因子水平检测（TNF-α、IL-1β等）
• 髓过氧化物酶（MPO）活性测定

模型特点
• 炎症峰值时间：给药后4-6小时
• 持续时间：24-48小时内可观察到明显炎症反应
• 适用研究：抗炎药物筛选、炎症机制研究等

注意事项
1. 严格控制环境温湿度（22±2℃）
2. 动物需提前适应环境至少3天
3. 操作时避免机械刺激影响耳部状态
4. 溶剂对照组应使用相同体积的PMA溶解溶剂
注：具体实验参数可根据研究目的调整，但需在文献中明确说明修改依据。建议初次建立模型时进行预实验确定最佳条件。

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Phorbol 12-myristate 13-acetate 可用于动物建模，构建湿疹样模型。 Phorbol 12-myristate 13-acetate (PMA) 是一种 PKC 激动剂，可恢复 5-癸酸 (5-HD) 引起的损伤因此，mitoKATP 的激活通过 PKC 保护了 SOD 和 MDA 中的线粒体功能[4]。

研究人员先前表明，过表达Thy-1抑制和敲低Thy-1增强内皮细胞迁移。在这里，研究人员使用phorpol -12-肉豆蔻酸酯-13-乙酸酯（PMA）作为Thy-1表达的诱诱剂来研究Thy-1上调的分子机制。数据显示，PMA处理后14-18小时和20-28小时内皮细胞中Thy-1 mRNA和蛋白水平分别升高。PMA处理32小时诱导Thy-1表达上调，抑制毛细血管样管形成和内皮细胞迁移。这些效应被Röttlerin （PKC-δ抑制剂）所消除，而Gö6976 （PKC-α/β抑制剂）则没有。此外，用Syk抑制剂Bay 61-3606或NF-κB抑制剂Bay 11-7082预处理可消除pma诱导的内皮细胞Thy-1上调和迁移抑制。通过斑马鱼模型，研究人员发现PMA通过PKC-δ介导的途径上调Thy-1并抑制血管生成。令人惊讶的是，他们发现短期（8-10小时）PMA治疗增强了内皮细胞的迁移。然而，在pma处理过表达thy -1的内皮细胞中没有观察到这种作用。综上所述，我们的研究结果表明，PMA最初增强了内皮细胞的迁移，随后激活PKC-δ/Syk/NF-κ b介导的通路，上调Thy-1，从而抑制内皮细胞的迁移。我们的结果也提示他们-1可能在终止血管生成中起作用。

单层培养的 αT3-1 和 LβT-2 细胞在 37°C、5% CO2 的加湿培养箱中的 DMEM 中生长。当向同一培养基中添加 0.1% FCS 时，血清饥饿持续 16 小时。然后，在指定的时间内，添加 GnRH 和 PMA。 αT3-1 细胞可以使用 jetPRIME 或 ExGen 500 暂时转染，而 LβT2 细胞只能使用 jetPRIME 转染试剂转染。在涉及显性失活 (DN) PKC 的实验中，将 1.5 μg p38α-GFP 或 3 μg DN-PKC 构建体与对照载体 pCDNA3 一起转染至 αT3-1 细胞（在 6 cm 平板中）。使用 4 μg p38α-GFP 与 9 μg DN-PKCs 构建体或对照载体 pCDNA3 组合进行 LβT2 细胞转染（在 10 cm 板中）。转染后约30小时，细胞血清饥饿（0.1% FCS）16小时。然后用 GnRH 或 PMA 刺激，用冰冷的 PBS 洗涤两次，用裂解缓冲液处理，然后进行一个冻融循环。收获细胞后，取上清液进行免疫沉淀实验，并在 4°C 下以 15,000 xg 离心 15 分钟。

Rats: Male Wistar rats weighing between 250 and 280 grams are used in all experiments. Seven groups of fifteen thirty-five Wistar rats are randomly assigned. (1) A 0.9% normal saline injection is administered to rats in the sham group (n = 21); (2) A 0.9% normal saline injection is administered to rats in the IR group (n = 21) 30 minutes prior to middle cerebral artery occlusion (MCAO); (3) A lateral cerebral ventricle injection of CBX (5 μg/mL×10 μL) is administered to rats in the Carbenoxolone (CBX) group (n = 21) 30 minutes prior to MCAO; (4) Rats in the Sch-6783 group (n = 21) receive a lateral cerebral ventricle injection of DZX (2 mM×30 μL) 30 minutes before MCAO; (5) Rats in the 5-HD group (n = 21) receive a lateral cerebral ventricle injection of 5-HD (100 mM×10 μL); (6) The rats in the DZX + Ro group (n = 15) receive a lateral cerebral ventricle injection of DZX, and after 10 min, Ro-31-8425 (400 μg/kg) is injected 15 minutes before MCAO; (7) Rats in the 5-HD+PMA group (n = 15) receive an intraperitoneal injection of PMA (200 μg/kg) following the injection of 5-HD and DZX.

Absorption, Distribution and Excretion
...Mouse skin localization expt...determined that at 3-6 hr after skin application /with tritiated PMA/ the keratin layer just above basal cells was highly labeled, & sebaceous glands & hair follicles were moderately labeled. After 48 hr there was still some labeling in sebaceous glands & hair follicles. Half-life of...promoter was close to 24 hr.

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Metabolism / Metabolites
...the major pathway in the metabolism of TPA is the hydrolysis of the two ester groups, ... in the rodent skin model all hydrolytic products lack tumor promoting activity, the major toxicological effect of TPA. The metabolic hydrolysis requires the activity of esterases, the activity of which differs between tissues and species. ... both ester groups of TPA can be hydrolysed in mouse skin and in cultured cells, giving rise to the monoesters 12-tetradecanoylphorbol and phorbol-13-acetate, as well as the product of complete hydrolysis, i.e. phorbol. Reduction of the keto group at C-3 was identified as a further metabolic pathway in mouse skin. ... Noteworthy, no other metabolites were detected in the microsomal incubations, suggesting that cytochrome 450-mediated oxidative metabolism is not involved in TPA metabolism. Ester group hydrolysis was also the only metabolic reaction observed in various cultured cells ... .
... the hydrolysis of TPA paralleled the loss of activity for induction of ornithine decarboxylase (ODC). As ODC is a marker for tumor promotion, these findings suggest that all three hydrolytic metabolites of TPA (the two monoesters and phorbol) are devoid of tumor promoting activity. Marked differences in the rate of hydrolysis of TPA and a structural analogue, phorbol-12,13-didecanoate (PDD) were observed between cultured fibroblasts from various animal species, suggesting that the hydrolytic metabolism of phorbol diesters depends on the cell type and on the chemical structure of the diester ... .
... the metabolism of radiolabeled TPA /was studied/ in the back skin of mice in vivo. In addition to hydrolytic metabolites, several novel lipophilic metabolites were detected and identified as TPA esterified with long chain fatty acids at the C-20 hydroxyl group. These TPA-20-acylates appeared to be devoid of tumor promoting activity but were partly hydrolysed back to TPA in mouse skin ... .
The few in vitro metabolism studies of TPA involving human cells indicate that many human cell lines in culture do not metabolize TPA to an appreciable extent ... .

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Biological Half-Life
... after /mouse/ skin application /with tritiated PMA/... Half-life of ... promoter was close to 24 hr.
A terminal half-life of 11 +/- 3.9 hours was calculated (from five infusions in four patients) ... .

Toxicity Summary
IDENTIFICATION AND USE: TPA is a colorless powder. It is used in cancer research to study mechanisms of tumor promotion, to screen for potential inhibiting agents, and as positive control for tumor promoting agents. It has been tested as experimental medication for the treatment of leukemias and lymphomas and other types of cancer. HUMAN EXPOSURE AND TOXICITY: TPA is a human platelet aggregating agent. TPA was used in clinical trials in humans suffering from recurrent malignancies, particularly hematological malignancies including severe forms of leukemia. The objective of this trial was the use of TPA as an agent to induce, at low doses,apoptosis and cell differentiation. The TPA application was based on current protocols for cytostatic agents, and involved 35 patients given a low dose constant rate infusion over a defined period. Various patients developed severe side effects following the treatment, such as transient fatigue, anemia, neutropenia and thrombocytopenia, mild dyspnea, nausea fever, rigor and cardiovascular effects with syncope and hypotension, but only one patient exhibited a tumor response, consisting in a reduction in mass dimensions. TPA is routinely used in human cell studies in vitro. ANIMAL STUDIES: TPA has been recognized as a tumor promoter in a mouse skin bioassay and in the mouse forestomach as well as in in vitro cell proliferation assays. However, there was no evidence for tumor-initiating properties of TPA. Tumor initiation and promotion was investigated in the epithelium of the forestomach of mice treated intragastrically with a single dose of 7,12-dimethylbenz [a]anthracene(DMBA) at 50 mg TPA/kg bw followed by repeated dosing (twice per week) for 35 weeks of TPA at 10 mg/kg bw. Forty-five out of 50 mice which received this treatment had tumors (papillomas) in the forestomach. There were no forestomach tumors noted for mice in the untreated control and the TPA-only groups, although in the DMBA-only group, papillomas were observed in the forestomach of 10 mice. TPA was not demonstrated to be a genotoxicant. Clastogenic, mutagenic and sister chromatid exchange-inducing effects of TPA have been shown in some experimental systems but are mediated by secondary products (possibly from arachidonic acid) formed by the cell, only under culture conditions with low antioxidant content in culture media and sera, in response to the tumor promoter. When tested in whole rat embryo culture, TPA exposure led to reduced prosencephalon, growth retardation and incomplete axial rotation in the body. An abundance of embryonic E-cadherin mRNA was found after culture. TPA is routinely used in animal cell studies in vitro.

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Interactions
To determine effect of interval between initiation & promotion & the effect of aging of mice in two-stage carcinogenesis, 20 ug 7,12-dimethylbenz(a)anthracene (initiator) was applied once only & 2.5 ug PMA was applied 3 times/wk to dorsal skin of 5 groups of female ICR/ha Swiss mice. For groups 1, 2, 3, 4 & 5, age (in wk) at primary treatment (initiator) was 6, 44, 56, 6 & 6 respectively; interval (in wk) to secondary treatment (promotor) was 2, 2, 2, 36 & 56 respectively; number of mice/group were 120, 20, 50, 35 & 35 respectively; % mice with papillomas was 100, 100, 56, 90 & 57 respectively; % mice with squamous carcinoma was 50, 30, 6, 25 & 11 respectively. Appropriate control groups consisted of one agent only given at various time intervals. Results show that skin carcinomas are induced whether interval between initiation & promotion is 2, 36 or 56 wk. Carcinoma incidences are significantly lower in groups 3 & 5 where secondary treatment was started when animals were 58 & 62 wk old. /From table/
Because endogenous proteases may play role in mechanism of action of tumor promotors, 3 known protease inhibitors were tested for ... inhibitory effects in two-stage carcinogenesis. Protease inhibitors ... tosyl chloromethyl ketone, tosyl phenylalanine chloromethyl ketone, & tosyl arginine methyl ester ... were applied to mouse ears after initiation with single dose of 7,12-dimethylbenz(a)anthracene followed by promotion with ... PMA. Inhibitors were applied 3 times/wk immediately after application of promoting agent. Protease inhibitors delayed appearance of 1st tumors, changed general pattern of rate of tumor appearance, & caused some decr in tumor incidences.
Low dosages of sulfur mustard, bis(beta-chloroethyl)sulfide completely inhibited two-stage carcinogenesis in mouse skin. 30 Female ICR/ha mice per group received either control applications or different combinations of initiator, promotor & inhibitor test compd for 400 days. 7,12-Dimethylbenz(a)anthracene (DMBA), 20 ug/0.1 mL acetone, applied once only was used as initiator. PMA, the promotor, was applied at 2.5 ug/0.1 mL acetone 3 times/wk. BCS, bis(beta-chloroethyl)sulfide, at 20 ug/0.1 mL acetone was applied beginning 14 days after initiator. DMBA + PMA + BCS (2 times/wk) produced papillomas in 1 mouse with 1st tumor occurring at 90 days; DMBA + PMA + BCS (3 times/wk) produced papillomas in 2 mice occurring at 209 days; DMBA + PMA only produced papillomas in 27 mice & squamous cell carcinomas in 16 occurring at 40 days; PMA alone produced papillomas in 4 mice occurring at 218 days; DMBA + BCS (2 times/wk) produced papillomas in 1 mouse at 385 days; DMBA + BCS (3 times/wk) produced no papillomas; BCS alone (3 times/wk) produced papillomas in 1 mouse at 323 days; DMBA + acetone only produced papillomas in 1 mouse at 219 days; no papillomas were observed in control group admin acetone alone or in group which did not receive either of the test compd. /From table/
The aim of the present study was to determine the effects of 12-O-tetradecanoylphorbol-13-acetate (TPA) and diethyldithiocarbamate (DDTC) alone or in combination on human pancreatic cancer cells cultured in vitro and grown as xenograft tumors in nude mice. Pancreatic cancer cells were treated with either DDTC or TPA alone, or in combination and the number of viable cells was then determined by trypan blue ecxlusion assay and the number of apoptotic cells was determined by morphological assessment by staining the cells with propidium iodide and examining them under a fluorescence microscope. Treatment with DDTC or TPA alone inhibited the growth and promoted the apoptosis of pancreatic cancer cells in a concentration-dependent manner. These effects were more prominent following treatment with TPA in combination with DDTC than following treatment with either agent alone in PANC-1 cells in monolayer cultures and in 3 dimensional (3D) cultures. The potent effects of the combination treatment on PANC-1 cells were associated with the inhibition of nuclear factor-kappaB (NF-kappaB) activation and the decreased expression of Bcl-2 induced by DDTC, as shown by NF-kappaB-dependent reporter gene expression assay and western blot analysis. Furthermore, treatment of nude mice with DDTC + TPA strongly inhibited the growth of PANC-1 xenograft tumors. The results of the present study indicate that the administration of TPA and DDTC in combination may be an effective strategy for inhibiting the growth of pancreatic cancer.
For more Interactions (Complete) data for 12-O-TETRADECANOYLPHORBOL-13-ACETATE (16 total), please visit the HSDB record page.

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Human Toxicity Values: LD50 Mouse iv 309 ug/kg

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Adverse Effects: Reproductive Toxin - A chemical that is toxic to the reproductive system, including defects in the progeny and injury to male or female reproductive function. Reproductive toxicity includes developmental effects. See Guidelines for Reproductive Toxicity Risk Assessment.

[1]. Autocrine and paracrine roles of sphingosine-1-phosphate. TRENDS in Endocrinology and Metabolism. Volume 18, Issue 8, October 2007, Pages 300-307. https://doi.org/10.1016/j.tem.2007.07.005

[2]. MPTP-Induced Depletion in Basolateral Amygdala via Decrease of D2R Activation Suppresses GABAA Receptors Expression and LTD Induction Leading to Anxiety-Like Behaviors. Front Mol Neurosci. 2017 Aug 7;10:247.

[3]. Differences in the state of differentiation of THP-1 cells induced by phorbol ester and 1,25-dihydroxyvitamin D3. J Leukoc Biol. 1996;59(4):555-561.

[4]. Differential roles of PKC isoforms (PKCs) and Ca2+ in GnRH and phorbol 12-myristate 13-acetate (PMA) stimulation of p38MAPK phosphorylation in immortalized gonadotrope cells. Mol Cell Endocrinol. 2017 Jan 5;439:141-154.

[5]. The phorbol 12-myristate-13-acetate differentiation protocol is critical to the interaction of THP-1 macrophages with Salmonella Typhimurium. PLoS One. 2018;13(3):e0193601.

[6]. Mechanism of Mitochondrial Connexin43's Protection of the Neurovascular Unit under Acute Cerebral Ischemia-Reperfusion Injury. Int J Mol Sci. 2016 May 5;17(5). pii: E679.

[7]. PMA inhibits endothelial cell migration through activating the PKC-δ/Syk/NF-κB-mediated up-regulation of Thy-1. Sci Rep. 2018 Nov 2;8(1):16247.

12-o-tetradecanoylphorbol-13-acetate appears as white crystals. (NTP, 1992)
Phorbol 13-acetate 12-myristate is a phorbol ester that is phorbol in which the hydroxy groups at the cyclopropane ring juction (position 13) and the adjacent carbon (position 12) have been converted into the corresponding acetate and myristate esters. It is a major active constituent of the seed oil of Croton tiglium. It has been used as a tumour promoting agent for skin carcinogenesis in rodents and is associated with increased cell proliferation of malignant cells. However its function is controversial since a decrease in cell proliferation has also been observed in several cancer cell types. It has a role as a protein kinase C agonist, an antineoplastic agent, a reactive oxygen species generator, a plant metabolite, a mitogen, a carcinogenic agent and an apoptosis inducer. It is an acetate ester, a tetradecanoate ester, a diester, a tertiary alpha-hydroxy ketone and a phorbol ester.
Phorbol 12-myristate 13-acetate diester is an inducer of neutrophil extracellular traps (NETs).
Phorbol 12-myristate 13-acetate has been reported in Iris tectorum, Phormidium tenue, and other organisms with data available.
Tetradecanoylphorbol Acetate is a phorbol ester with potential antineoplastic effects. Tetradecanoylphorbol acetate (TPA) induces maturation and differentiation of hematopoietic cell lines, including leukemic cells. This agent may induce gene expression and protein kinase C (PKC) activity. In addition to potential antineoplastic effects, TPA may exhibit tumor promoting activity. (NCI04)
Phorbol Ester is polycyclic compound isolated from croton oil in which two hydroxyl groups on neighboring carbon atoms are esterified to fatty acids. The commonest of these derivatives is phorbol myristoyl acetate (PMA). Potent co carcinogens or tumor promotors, they are diacyl glycerol analogs and activate protein kinase C irreversibly.
A phorbol ester found in CROTON OIL with very effective tumor promoting activity. It stimulates the synthesis of both DNA and RNA.

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Mechanism of Action
The tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA), has a differential role on the regulation of the cell cycle in a variety of tumor cells. The mechanism between TPA and the cell cycle in breast cancer is not fully understood. Therefore, we investigated the regulatory mechanism of TPA on control of the cell cycle of breast cancer cells. Our results showed that TPA increased the level of p21 expression in MCF-7 cells with wild-type p53 and MDA-MB-231 cells with mutant p53 in a dose-dependent manner. In contrast, TPA decreased the expression of p53 in MCF-7 cells, but did not affect MDA-MB-231 cells. We next examined the regulatory mechanism of TPA on p21 and p53 expression. Our results showed that the TPA-induced up-regulation of p21 and down-regulation of p53 was reversed by UO126 (a MEK1/2 inhibitor), but not by SP600125 (a JNK inhibitor) or SB203580 (a p38 inhibitor), although TPA increased the phosphorylation of ERK and JNK in MCF-7 cells. In addition, the TPA-induced arrest of the G2/M phase was also recovered by UO126 treatment. To confirm the expression of p21 through the MEK/ERK pathway, cells were transfected with constitutively active (CA)-MEK adenovirus. Our results showed that the expression of p21 was significantly increased by CA-MEK overexpression. Taken together, we suggest that TPA reciprocally regulates the level of p21 and p53 expression via a MEK/ERK-dependent pathway. The up-regulation of p21 in response to TPA is mediated through a p53-independent mechanism in breast cancer cells.

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Therapeutic Uses
/CLINICAL TRIALS/ ClinicalTrials.gov is a registry and results database of publicly and privately supported clinical studies of human participants conducted around the world. The Web site is maintained by the National Library of Medicine (NLM) and the National Institutes of Health (NIH). Each ClinicalTrials.gov record presents summary information about a study protocol and includes the following: Disease or condition; Intervention (for example, the medical product, behavior, or procedure being studied); Title, description, and design of the study; Requirements for participation (eligibility criteria); Locations where the study is being conducted; Contact information for the study locations; and Links to relevant information on other health Web sites, such as NLM's MedlinePlus for patient health information and PubMed for citations and abstracts for scholarly articles in the field of medicine. 12-O-Tetradecanoylphorbol-13-acetate is included in the database.
/EXPL THER/ Phorbol esters activate protein kinase C and modulate a variety of downstream cell signaling pathways. 12-O-tetradecanoylphorbol-13-acetate (TPA) is a phorbol ester that induces differentiation or apoptosis in a variety of cell lines at low concentrations. A phase I dose escalation trial of TPA was undertaken for patients with relapsed or refractory malignancies. The starting dose was 0.063 mg/sq m and most patients were treated with an intravenous infusion of TPA on days 1-5 and 8-12 followed by a 2-week rest period prior to retreatment. Thirty-five patients were treated. A biological assay was used to monitor levels of TPA-like activity in the blood after treatment. Serious adverse events included individual episodes of gross hematuria, a grand mal seizure, syncope, and hypotension. Many patients had transient fatigue, mild dyspnea, fever, rigors, and muscular aches shortly after the infusion. Dose-limiting toxicities included syncope and hypotension at a dose of 0.188 mg/sq m. Only a single patient had evidence of tumor response. These studies establish 0.125 mg/sq m as the maximally tolerated dose when TPA is administered on this schedule.

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We examined the role of PKCs and Ca2+ in GnRH-stimulated p38MAPK phosphorylation in the gonadotrope derived αT3-1 and LβT2 cell lines. GnRH induced a slow and rapid increase in p38MAPK phosphorylation in αT3-1 and LβT2 cells respectively, while PMA gave a slow response. The use of dominant negatives for PKCs and peptide inhibitors for the receptors for activated C kinase (RACKs), has revealed differential role for PKCα, PKCβII, PKCδ and PKCε in p38MAPK phosphorylation in a ligand-and cell context-dependent manner. The paradoxical findings that PKCs activated by GnRH and PMA play a differential role in p38MAPK phosphorylation may be explained by differential localization of the PKCs. Basal, GnRH- and PMA- stimulation of p38MAPK phosphorylation in αT3-1 cells is mediated by Ca2+ influx via voltage-gated Ca2+ channels and Ca2+ mobilization, while in the differentiated LβT2 gonadotrope cells it is mediated only by Ca2+ mobilization. p38MAPK resides in the cell membrane and is relocated to the nucleus by GnRH (∼5 min). Thus, we have identified the PKCs and the Ca2+ pools involved in GnRH stimulated p38MAPK phosphorylation.[4]

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THP-1 cells differentiated with phorbol 12-myristate 13-acetate (PMA) are widely used as a model for function and biology of human macrophages. However, the conditions used for differentiation, particularly the concentration of PMA and the duration of treatment, vary widely. Here we compare several differentiation conditions and compare the ability of THP-1 macrophages to interact with the facultative intracellular pathogen Salmonella enterica serovar Typhimurium. The results show that THP-1 macrophages differentiated in high concentrations of PMA rapidly died following infection whereas those differentiated in low concentrations of PMA survived and were able to control the intracellular bacteria similar to primary human macrophages.[5]

C36H56O8

616.8251

616.397

C, 70.10; H, 9.15; O, 20.75

16561-29-8

27924

White to off-white solid powder

1.2±0.1 g/cm3

698.1±55.0 °C at 760 mmHg

162 °F (NTP, 1992)
50-70 °C (melting pt-freezing pt)

208.1±25.0 °C

0.0±5.0 mmHg at 25°C

1.553

7.71

130.36

3

8

17

44

1150

8

O(C(C([H])([H])[H])=O)[C@@]12[C@@]([H])([C@@]([H])(C([H])([H])[H])[C@@]3([C@]4([H])C([H])=C(C([H])([H])[H])C([C@]4(C([H])([H])C(C([H])([H])O[H])=C([H])[C@@]3([H])[C@]1([H])C2(C([H])([H])[H])C([H])([H])[H])O[H])=O)O[H])OC(C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H])=O

PHEDXBVPIONUQT-RGYGYFBISA-N

InChI=1S/C36H56O8/c1-7-8-9-10-11-12-13-14-15-16-17-18-29(39)43-32-24(3)35(42)27(30-33(5,6)36(30,32)44-25(4)38)20-26(22-37)21-34(41)28(35)19-23(2)31(34)40/h19-20,24,27-28,30,32,37,41-42H,7-18,21-22H2,1-6H3/t24-,27+,28-,30-,32-,34-,35-,36-/m1/s1

[(1S,2S,6R,10S,11R,13S,14R,15R)-13-acetyloxy-1,6-dihydroxy-8-(hydroxymethyl)-4,12,12,15-tetramethyl-5-oxo-14-tetracyclo[8.5.0.02,6.011,13]pentadeca-3,8-dienyl] tetradecanoate

TPA; NSC262244; PD616; PMA; Phorbol myristate acetate; NSC 262244; Phorbol 12-myristate 13-acetate; 16561-29-8; phorbol-12-myristate-13-acetate; 12-O-Tetradecanoylphorbol-13-acetate; 12-O-Tetradecanoylphorbol 13-acetate; Tetradecanoylphorbol acetate; Phorbol ester; Factor A1; NSC-262244; PD-616; RP-323; PD 616; RP 323; PMA; RP323

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)

DMSO: ~100 mg/mL (~162.1 mM)
Ethanol: ~100 mg/mL (~162.1 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (4.05 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 25.0 mg/mL澄清DMSO储备液加入到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。

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配方 2 中的溶解度: 2.5 mg/mL (4.05 mM) 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), 悬浊液; 超声助溶。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 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。

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配方 3 中的溶解度: ≥ 2.5 mg/mL (4.05 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 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: ≥ 2.5 mg/mL (4.05 mM) (饱和度未知) in 10% EtOH + 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 25.0 mg/mL 澄清 EtOH 储备液加入400 μL PEG300 中，混匀；再向上述溶液中加入50 μL Tween-80，混匀；然后加入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。

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配方 5 中的溶解度: 2.5 mg/mL (4.05 mM) in 10% EtOH + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
例如，若需制备1 mL的工作液，可将100μL 25.0mg/mL澄清EtOH储备液加入到900μL 20％SBE-β-CD生理盐水中，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。

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配方 6 中的溶解度: ≥ 2.5 mg/mL (4.05 mM) (饱和度未知) in 10% EtOH + 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 25.0 mg/mL 澄清乙醇储备液加入到 900 μL 玉米油中并混合均匀。

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配方 7 中的溶解度: 5%DMSO + Corn oil: 5.0mg/ml (8.11mM)

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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。