CK2 ( IC50 = 40 nM )

体外活性：鞣花酸是一种有效的天然 CK2 抑制剂，IC50 为 40 nM，Ki 为 20 nM。鞣花酸还阻断其他激酶，如 LYN、PKA、SYK、GSK3、FGR 和 CK1，IC50 分别为 2.9、3.5、4.3、7.5、9.4 和 13.0 μM，对 DYRK1a、CSK、NPM- 无明显影响。 ALK、RET 和 FLT3 (IC50 > 40 μM)。鞣花酸 (5-100 μM) 对 Karpas299、SUDHL1、SR786 和 FE-PD 细胞系具有抑制活性[1]。鞣花酸 (10 μM) 在放射治疗后对 MCF-7 细胞表现出细胞毒性作用。鞣花酸 (10 μM) 与辐照 (IR) 结合显着降低了 MCF-7 细胞形成集落的能力，这与单独处理相同。鞣花酸与 IR 还可诱导细胞凋亡，并促进 MCF-7 细胞中促凋亡 Bax 的上调和 Bcl-2 的下调。激酶测定：CK2 和 CK1 磷酸化测定在 37°C 下进行，每种抑制剂（鞣花酸）的量不断增加，最终体积为 25 µL，含有 50 mM Tris-HCl pH 7.5、100 mM NaCl，12 mM MgCl2，0.02 mM [33P-ATP] (500-1000 cpm/pmol)，除非另有说明。 CK2 和 CK1 的可磷酸化底物分别是合成肽底物 RRRADDSDDDDD (100 µM) 和 RRKHAAIGDDDDAYSITA (200 µM)。反应从添加激酶开始，10 分钟后停止。在将等分试样点样到磷酸纤维素滤膜上之前，添加 5 µL 0.5 M 正磷酸。对放射性标记样品进行 SDS-PAGE 后，用 75 mM 磷酸底物清洗过滤器。 DYRK1A，对肽 RRRFRPASPLRGPPK 进行测定，并确定酪氨酸激酶活性。 细胞测定：通过 MTT 测定测量 ALCL 细胞活力。简而言之，在添加鞣花酸前 12 小时将 0.1 × 105 个细胞接种到 96 孔微量培养板上。在标准组织培养条件下，在存在或不存在药物（鞣花酸）的情况下，细胞在 200 µL 完全 RPMI-1640 培养基中生长 48 小时。然后将 20 µL MTT 溶液 (5 mg/mL) 添加到细胞悬浮液中 4 小时。将细胞内甲臜晶体溶解在 150 µL DMSO 中，在分光光度计上于 540 nm 处测量的光密度代表一式三份培养物的平均值 (± SD)。

鞣花酸（EA；10 mg/kg/天；口服，14 天）可显着降低大鼠脑中 MDA 含量 17%，并使脑 TNF-α 水平降低 42%。鞣花酸显着增加大脑中减少的 5-HT（39%）、多巴胺（DA，71%）和去甲肾上腺素（NE，77%）的含量。鞣花酸（10 mg/kg，口服，14 天）可减少多柔比星引起的大鼠组织病理学变化。

CK2 和 CK1 磷酸化测试在 37°C 下进行，最终体积为 25 µL，其中含有 50 mM、pH 7.5 Tris-HCl、100 mM NaCl、12 mM MgCl_{，每种抑制剂（鞣花酸）的浓度不断增加2} 和 0.02 mM [³³P-ATP] (500-1000 cpm/pmol)，除非另有说明。对于 CK1 和 CK2，可磷酸化底物分别为 RRKHAAIGDDDDAYSITA (200 µM) 和 RRRADDSDDDDD (100 µM)，合成肽底物。首先加入激酶，10分钟后，加入5μL 0.5M正磷酸终止反应。这是在将等分试样置于磷酸纤维素过滤器上之前完成的。通过 SDS-PAGE 分离放射性标记的样品后，在 75 mM 磷酸底物中清洗过滤器。 DYRK1A 的酪氨酸激酶活性使用肽 RRRFRPASPLRGPPK 进行测量[1]。

MTT 测定用于确定 ALCL 细胞活力。简而言之，在添加鞣花酸前 12 小时，将 0.1 × 10⁵ 细胞接种到 96 孔微量培养板上。按照标准组织培养程序，将细胞在 200 µL 全 RPMI-1640 培养基中培养 48 小时，无论是否添加药物（鞣花酸）。接下来，将 20 µL MTT 溶液 (5 mg/mL) 添加到细胞悬浮液中，并静置 4 小时。使用 150 µL DMSO 溶解细胞内甲臜晶体，使用分光光度计在 540 nm 处测定的光密度代表三个重复培养物的平均值 (± SD)[1]。

Fifty adult male Sprague-Dawley rats are split into five groups at random, which are as follows: As a vehicle and standard control, Group (1) is given oral corn oil. Doxorubicin (DOX) injections (5 mg/kg, i.p.) are given to Group (2) twice a week for a duration of 14 days. For a duration of 14 days, Group 3 is administered Ellagic acid (10 mg/kg, p.o.; daily) and DOX (5 mg/kg, i.p.) twice a week. For 14 days, rosmarinic acid (RA; 75 mg/kg, p.o.; daily) and DOX (5 mg/kg, i.p.) are given to Group 4. For a duration of 14 days, Group 5 is administered Ellagic acid (10 mg/kg, p.o.; daily) along with RA (75 mg/kg, p.o.; daily) and a DOX injection (5 mg/kg, i.p.) twice a week[2].

Absorption, Distribution and Excretion
After oral consumption, ellagic acid reaches maximum concentrations in about 1 hour.
Ellagic acid is eliminated from the body in about 4 hours.
The present study was initiated to determine ... the distribution of (14)C-ellagic acid (EA) and (3)H-N-methyl-N-nitrosourea (MNU) in the rat whole embryo culture model system ... (14)C-EA (50 uM for 2 hr, known embryoprotective concentration; no MNU added) was used to demonstrate access of EA to the embryo within the 2 hr exposure period. The majority of EA (99.5%) remained in the media while tissue concentrations of 57.0 and 47.9 pmol/mg were attained in the yolk sacs and embryos, respectively.
Ellagic acid (EA), derived from fruit ellagitannins, is known to be antimutagenic and anticarcinogenic in various animal tumor models. In this study, EA at a dose of 4 g/kg diet inhibited multiplicity of tumors induced by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in A/J mice by 54%. This inhibition was dose related between 0.06 and 4.0 g/kg diet. In contrast, two related compounds, esculin and esculetin, had no effect on lung tumorigenesis. The biodistribution of ellagic acid (EA) /in A/J mice/ was studied as a function of dose and time after gavage of EA. The levels of EA in the lung were directly proportional to the dose of EA between 0.2 and 2.0 mmol. The maximum level of EA, corresponding to 21.3 nmol/g, was observed 30 minutes after gavage with 2.0 mmol of EA/kg body wt, which corresponds to only 70 ppm of the administered dose. The levels in liver tissues were 10-fold lower and reached a maximum 30 minutes after gavage. At this interval, the blood level of EA was 1 nmol/mL. The inclusion of EA in cyclodextrin doubles the level of EA in lung tissues. These results demonstrate that EA localizes preferentially in lung tissues ...
... Ellagic acid (EA), a dietary antioxidant associated with poor biopharmaceutical properties, was encapsulated into poly(lactide-co-glycolide) (PLGA) and polycaprolactone (PCL) nanoparticles to improve oral bioavailability ... The antioxidant potential of the didodecyldimethyl ammonium bromide (DMAB)-stabilized nanoparticulate formulations was evaluated against cyclosporine A (CyA)-induced nephrotoxicity in rats ... From in situ permeation studies in rats, it was evident that intestinal uptake of EA as DMAB-stabilized nanoparticles was significantly higher as compared to the sodium carboxymethyl cellulose suspension and the polyvinyl alcohol (PVA)-stabilized particles. EA and EA nanoparticles were able to prevent the CyA-induced nephrotoxicity in rats as evident by biochemical parameters as well as kidney histopathology.

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
Pomegranate juice contains ellagitannins and numerous polyphenols such as delphinidin, cyanidin, pelargonidin, punicalin, pedunculagin, punicalagin, gallagic, and ellagic acid as well as vitamin C and B vitamins. Administration of pomegranate juice to nursing mothers result in a change in the microbiota of breastmilk and infant stools and an increase in antioxidant content of breastmilk and infant urine. A low-quality study found that administration of pomegranate juice concentrate to nursing mothers of infants with hyperbilirubinemia hastened the infant’s improvement with phototherapy treatment. No adverse reactions to maternal pomegranate ingestion in breastfed infants have been observed.
◉ Effects in Breastfed Infants
Twelve exclusively breastfeeding mothers of full-term infants consumed 8 fluidounces of pomegranate juice daily for 2 weeks. After 2 weeks, the infant stool samples had significant increases in the abundance of bacteria from the phylum Firmicutes (genus Lachnoclostridium and Staphylococcus). Infants whose mothers’ milk contained urolithin B-glucuronide (metabotype B) showed significant differences in 4 bacteria, with 2 bacteria from the phyla Firmicutes (genus Veillonella) and Bacteroidetes (genus Bacteroides and Parabacteroides) and Bifidobacterium. Infant stool Blautia was positively corelated with breastmilk and plasma urolithin B-glucuronide and infant stool Enterococcus was inversely related to breastmilk maternal plasma urolithin B-glucuronide, and maternal plasma urolithin B-glucuronide and dimethyl ellagic acid glucuronide.
An open-label, nonblinded study randomized mothers of newborn infants to receive either concentrated pomegranate juice 15 mL 3 times daily or nothing (i.e., no placebo control). There were 43 mother-infant pairs in each group. The concentrate was made from the juice of fresh pomegranates concentrated at a warm temperature to a Brix of 60%. The total phenol content was 13.56 mg/g and the total flavonoid was 1.39 mg/g. All of the infants were over 72 hours old, had a gestational age over 37 weeks and birth weight over 2500 grams. Both groups were being treated with phototherapy for hyperbilirubinemia (total serum bilirubin level over 15 mg/dL). There was a greater decrease in bilirubin levels in the pomegranate group at 48 and 72 hours after the start of phototherapy and at 48 hours after discharge. In addition, the mean duration of phototherapy in the pomegranate group was significantly shorter (52 hours compared to 65 hours) than in the no-pomegranate group. All infants in the pomegranate group were discharged by 96 hours after the start of treatment compared to 114 hours in the no-pomegranate group. No side effects were observed in the infants whose mothers received pomegranate juice concentrate.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

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Interactions
Naturally occurring plant phenols with antimutagenic and anticarcinogenic activities were tested for their abilities to inhibit the biochemical and biological effects of the potent tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA) in mouse epidermis in vivo. When applied topically to mouse skin, tannic acid (TA), ellagic acid, and several gallic acid derivatives all inhibit TPA-induced ornithine decarboxylase activity, hydroperoxide production, and DNA synthesis, three biochemical markers of skin tumor promotion. Moreover, in the two-step initiation-promotion protocol, the same phenolic compounds also inhibit the incidence and yield of skin tumors promoted by TPA.
Resveratrol is shown to have a synergistic effect in vitro with both quercetin and ellagic acid for apoptosis ...
... Ellagic acid (EA) at a dose of 4 g/kg diet inhibited multiplicity of tumors induced by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in A/J mice by 54%. This inhibition was dose related between 0.06 and 4.0 g/kg diet ...
When administered in a semi-purified diet at concentrations of 0.4 and 4 g/kg, ellagic acid (EA) produced a significant (21 to 55%) decrease in the average number of N-nitrosobenzylmethylamine (NBMA)-induced esophageal tumors after 20 and 27 weeks of the bioassay. EA exhibited inhibitory effects toward preneoplastic lesions as well as neoplastic lesions. Tumors were not observed in vehicle-control rats or in rats that received EA alone.
For more Interactions (Complete) data for ELLAGIC ACID (6 total), please visit the HSDB record page.

[1]. Identification of ellagic acid as potent inhibitor of protein kinase CK2: a successful example of a virtual screening application. J Med Chem. 2006 Apr 20;49(8):2363-6.

[2]. Prophylactic effects of ellagic acid and rosmarinic acid on doxorubicin-induced neurotoxicity in rats. J Biochem Mol Toxicol. 2017 Dec;31(12).

[3]. Ellagic Acid Enhances Apoptotic Sensitivity of Breast Cancer Cells to γ-Radiation. Nutr Cancer. 2017 Aug-Sep;69(6):904-910.

[4]. Discovery of ellagic acid as a competitive inhibitor of Src homology phosphotyrosyl phosphatase 2 (SHP2) for cancer treatment: In vitro and in silico study. Int J Biol Macromol. 2023 Nov 5;254(Pt 2):127845.

Therapeutic Uses
/EXPL THER/ Italian researchers found that ellagic acid seemed to reduce the side effects of chemotherapy in men with advanced prostate cancer, although it did not help slow disease progression or improve survival.
/EXPL THER/ Ellagic acid seems to have some anti-cancer properties. It can act as an antioxidant, and has been found to cause apoptosis (cell death) in cancer cells in the lab. In other lab studies, ellagic acid seems to reduce the effect of estrogen in promoting growth of breast cancer cells in tissue cultures. There are also reports that it may help the liver to break down or remove some cancer-causing substances from the blood. Some supporters have claimed these results mean that ellagic acid can prevent or treat cancer in humans. This has not been proven. Unfortunately, many substances showing promise against cancer in lab and animal studies have not been found to be useful in people. Ellagic acid has also been said to reduce heart disease, birth defects, liver problems, and to promote wound healing. The available scientific research does not support these claims at this time.
/EXPL THER/ ... Ellagic acid (EA) at a dose of 4 g/kg diet inhibited multiplicity of tumors induced by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in A/J mice by 54%. This inhibition was dose related between 0.06 and 4.0 g/kg diet ...
/EXPL THER/ When administered in a semi-purified diet at concentrations of 0.4 and 4 g/kg, ellagic acid (EA) produced a significant (21 to 55%) decrease in the average number of N-nitrosobenzylmethylamine (NBMA)-induced esophageal tumors after 20 and 27 weeks of the bioassay. EA exhibited inhibitory effects toward preneoplastic lesions as well as neoplastic lesions. Tumors were not observed in vehicle-control rats or in rats that received EA alone.
/EXPL THER/ Ellagic acid inhibited lung tumorigenesis induced by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in A/J mice. This inhibition was related to the logarithm of the dose of ellagic acid added to the diet. The biodistribution of ellagic acid was studied in mice gavaged with ellagic acid. Pulmonary levels of ellagic acid were directly proportional to the dose of ellagic acid between 0.2 and 2.0 mmol/kg ...

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Drug Warnings
Ellagic acid is available in supplement form, but it has not been tested for safety. Some reports indicate it may affect certain enzymes in the liver, which could alter levels of some drugs in the body. For this reason, people taking medicines or other dietary supplements should talk with their doctors or pharmacists about all their medicines and supplements before taking ellagic acid. The raspberry leaf, or preparations made from it, should be used with caution during pregnancy because they may initiate labor.

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Pharmacodynamics
Ellagic acid's therapeutic action mostly involves antioxidant and anti-proliferative/anti-cancer effects.

C14H6O8

302.19

302.006

C, 55.64; H, 2.00; O, 42.35

476-66-4

133039-73-3;314041-08-2

5281855

Light yellow to khaki solid powder

2.1±0.1 g/cm3

796.5±60.0 °C at 760 mmHg

≥350 °C

310.1±26.4 °C

0.0±2.9 mmHg at 25°C

1.895

0.52

141.34

4

8

0

22

475

0

O1C(C2=C([H])C(=C(C3=C2C2=C1C(=C(C([H])=C2C(=O)O3)O[H])O[H])O[H])O[H])=O

AFSDNFLWKVMVRB-UHFFFAOYSA-N

InChI=1S/C14H6O8/c15-5-1-3-7-8-4(14(20)22-11(7)9(5)17)2-6(16)10(18)12(8)21-13(3)19/h1-2,15-18H

6,7,13,14-tetrahydroxy-2,9-dioxatetracyclo[6.6.2.04,16.011,15]hexadeca-1(15),4,6,8(16),11,13-hexaene-3,10-dione

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)

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。