Natural polyphenol; anticancer

叶子中(-)-没食子酸酯的含量极少，并且不随阶段的不同而变化[1]。当 (−)-表没食子儿茶素没食子酸酯和活性儿茶素 ((−)-表没食子儿茶素没食子酸酯) 组合时，它们具有协同作用，可阻止 PC-9 细胞发育并诱导细胞凋亡。 (−)-没食子儿茶素没食子酸酯抑制 α-葡萄糖苷酶和 DPPH，IC50 值分别为 30.2 μM 和 12.2 μg/mL [2]。

α-葡萄糖苷酶抑制试验[2]
该测定在96孔微量滴定板上按照文献中描述的程序进行（杨等人，2016a，杨等人，2016.b）。简而言之，将测试化合物溶解在二甲亚砜（DMSO）中，达到六个系列浓度。α-葡萄糖苷酶和p-NPG分别溶解在pH 6.8的60 mM磷酸钠缓冲液中，浓度分别为0.5 U/mL和5 mM。提出了四种解决方案。供试品溶液含有112μL缓冲液、20μL酶和8μL化合物。测试库溶液含有112μL缓冲液和8μL测试化合物。阴性对照溶液含有112μL缓冲液、20μL酶和8μL DMSO。阴性空白溶液含有132μL缓冲液和8μL DMSO。阿卡波糖用作阳性对照。小心摇动平板，使溶液充分混合，并在37°C下保持15分钟。加入总共20μL的p-NPG以淬灭反应。通过测量405nm处的OD值来定量α-葡萄糖苷酶从p-NPG水解的对硝基苯酚的量。

---

DPPH自由基清除试验[2]
根据先前描述的方法测量化合物的DPPH自由基清除活性。简而言之，将20μL不同浓度的测试样品与180μL DPPH溶液在黑暗中混合30分钟。然后，在微孔板读数器上测量DPPH在517 nm处的吸光度变化。DMSO用作阴性对照。结果表示为相对于含有DMSO代替样品的对照的抑制百分比，以及半最大抑制浓度（IC50值，μg/mL）。

细胞毒性试验[2]
通过测量细胞将MTT代谢为紫色甲赞染料的能力来检查细胞存活率（Zhou等人，2015）。人肝癌癌症HepG2细胞在添加有10%胎牛血清、100单位mL−1青霉素和50单位mL−2链霉素的DMEM培养基中，在37°C下，在含5%CO2气氛的湿润培养箱中维持。将细胞接种在96孔组织培养板中24小时，然后与不同浓度的受试化合物一起孵育72小时。孵育后，加入5 mg/mL PBS中的25μL MTT并孵育4小时。吸出培养基，用150μL二甲亚砜（DMSO）代替以溶解甲胺盐。使用微孔板分光光度计在570nm处测量反映细胞生长状况的甲赞溶液的颜色强度。

mouse LD50 oral >1 gm/kg Japanese Kokai Tokyo Koho Patents., #93-944

[1]. Accumulation of catechins and expression of catechin synthetic genes in Camellia sinensis at different developmental stages. Bot Stud. 2016 Dec;57(1):31.

[2]. C-geranylated flavanones from YingDe black tea and their antioxidant and α-glucosidase inhibition activities. Food Chem. 2017 Nov 15;235:227-233.

(-)-gallocatechin gallate is a gallate ester obtained by formal condensation of the carboxy group of gallic acid with the (3R)-hydroxy group of (-)-gallocatechin. A natural product found in found in green tea. It has a role as an EC 3.4.22.69 (SARS coronavirus main proteinase) inhibitor, a human xenobiotic metabolite, an antineoplastic agent and a plant metabolite. It is a gallate ester, a polyphenol and a catechin. It is functionally related to a (-)-gallocatechin and a gallic acid. It is an enantiomer of a (+)-gallocatechin gallate.
(-)-Gallocatechin gallate has been reported in Camellia sinensis, Potentilla erecta, and other organisms with data available.

---

Background: Catechins are the main polyphenol compounds in tea (Camellia sinensis). To understand the relationship between gene expression and product accumulation, the levels of catechins and relative expressions of key genes in tea leaves of different developmental stages were analyzed.
Results: The amounts of catechins differed significantly in leaves of different stages, except for gallocatechin gallate. Close correlations between the expression of synthesis genes and the accumulation of catechins were identified. Correlation analysis showed that the expressions of chalcone synthase 1, chalcone synthase 3, anthocyanidin reductase 1, anthocyanidin reductase 2 and leucoanthocyanidin reductase genes were significantly and positively correlated with total catechin contents, suggesting their expression may largely affect total catechin accumulation. Anthocyanidin synthase was significantly correlated with catechin. While both ANRs and LAR were significantly and positively correlated with the contents of (-)-epigallocatechin gallate and (-)-epicatechin gallate.
Conclusion: Our results suggest synergistic changes between the expression of synthetic genes and the accumulation of catechins. Based on our findings, anthocyanidin synthase may regulate earlier steps in the conversion of catechin, while the anthocyanidin reductase and leucoanthocyanidin reductase genes may both play important roles in the biosynthesis of galloylated catechins.[1]

---

YingDe black tea is produced from crude tea prepared from leaves of Camellia sinensis var. assamica. In this work, we isolated and identified five novel flavanones, namely, amelliaone A-E (1-5), along with seven known compounds 6-12 from the ethanol extract of YingDe black tea. The structures of these five novel phenolic compounds were determined using extensive 1D and 2D nuclear magnetic resonance spectroscopy experiments. The compounds were further evaluated for antioxidant, α-glucosidase inhibitory, and cytotoxic activities. Compound 1 exhibited higher α-glucosidase inhibitory activity with a half-maximum inhibitory concentration value (IC50) of 10.2µM compared with acarbose (18.2µM).[2]

C22H18O11

458.375

458.084

C, 57.65; H, 3.96; O, 38.40

4233-96-9

(-)-Epigallocatechin Gallate;989-51-5

199472

Typically exists as solid at room temperature

1.9±0.1 g/cm3

909.1±65.0 °C at 760 mmHg

320.0±27.8 °C

0.0±0.3 mmHg at 25°C

1.857

Tea

2.08

197.37

8

11

4

33

667

2

O1C2=C([H])C(=C([H])C(=C2C([H])([H])[C@]([H])([C@]1([H])C1C([H])=C(C(=C(C=1[H])O[H])O[H])O[H])OC(C1C([H])=C(C(=C(C=1[H])O[H])O[H])O[H])=O)O[H])O[H]

WMBWREPUVVBILR-NQIIRXRSSA-N

InChI=1S/C22H18O11/c23-10-5-12(24)11-7-18(33-22(31)9-3-15(27)20(30)16(28)4-9)21(32-17(11)6-10)8-1-13(25)19(29)14(26)2-8/h1-6,18,21,23-30H,7H2/t18-,21+/m1/s1

[(2S,3R)-5,7-dihydroxy-2-(3,4,5-trihydroxyphenyl)-3,4-dihydro-2H-chromen-3-yl] 3,4,5-trihydroxybenzoate

CCRIS-9286; CCRIS 9286; (-)-Gallocatechin gallate; 4233-96-9; (-)-Gallocatechol gallate; (2S,3R)-5,7-dihydroxy-2-(3,4,5-trihydroxyphenyl)chroman-3-yl 3,4,5-trihydroxybenzoate; [(2S,3R)-5,7-dihydroxy-2-(3,4,5-trihydroxyphenyl)-3,4-dihydro-2H-chromen-3-yl] 3,4,5-trihydroxybenzoate; Gallocatechin gallate, (-)-; (2S,3R)-2-(3,4,5-Trihydroxyphenyl)-3,4-dihydro-1(2H)-benzopyran-3,5,7-triol 3-(3,4,5-trihydroxybenzoate); MFCD00214298; CCRIS9286; (-)-gallocatechin-3-O-gallate; (-)-Gallocatechol gallate; Gallocatechin gallate

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)

DMSO : ≥ 100 mg/mL (~218.16 mM)

配方 1 中的溶解度: ≥ 2.08 mg/mL (4.54 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 (4.54 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 (4.54 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。