Natural flavonoid in green tea; COX-1/cyclooxygenase-1

At 70 μg/mL and an IC50 of 1.4 μM, (+)-catechin demonstrates >95% inhibitory action against cyclooxygenase-1 (COX-1) [1]. Following a 24-hour treatment with (+)-catechin, a dose-dependent decrease in color was noted. At the highest concentration of (+)-catechin tested (160 μg/mL), 54.76% of cells perished, with an IC50 of (+)-catechin at 127.62 μg/mL. Treatment of MCF-7 cells with (+)-catechin resulted in an increase in the induction of apoptosis that was dose- and time-dependent. After 24 hours, 40.7% and 41.16% of the cells treated with 150 μg/mL and 300 μg/mL (+)-catechin experienced apoptosis, in comparison to the control cells. Following a 24-hour treatment with 150 μg/mL (+)-catechin, MCF-7 cells exhibited 5.81, 1.42, 3.29, and 2.68-fold increases in Caspase-3, -8, and -9 expression levels, respectively. Comparison with untreated control cell levels [2].

Although the difference was not statistically significant, animals treated with (+)-catechin at the lowest tested dose (i.e., 50 mg/kg, po) explored unfamiliar targets substantially more frequently in choice trials. Time-induced episodic memory losses are prevented by (+)-catechin in a dose-dependent manner; 200 mg/kg orally is the most efficacious dose. Treatment with (+)-catechin prevented an increase in MPO levels (21.98±9.44% in the hippocampus and 36.76±4.39% in the frontal cortex) compared to the DOX treatment group alone [3].

Natural flavonoid in green tea In vitro studies[2]
IMR-32 cell line is male Caucasian derived neuroblastoma cell line. It was procured from NCCS, subcultured in DMEM medium with 10% of fetal bovine serum. The cells were used for cell viability and cyto-protection studies. The treatments of Catechin and DOX in the neuroprotection study were simultaneous (without changing the medium), where Catechin was added 1 h prior to DOX addition.

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MTT assay in undifferentiated IMR-32 cells[2]
Five thousand cells per well were seeded in microplate consisting of 50 µl of medium and incubated for 24 h. After 24 h, 50 µl of Catechin was added in a concentration ranging from 31.23 to 250 µg/ml in the wells for one hour. After that 50 µl of DOX (1 or 2 µg/ml) was added and incubated for 24 h. Followed by that 50 µl of MTT (2 mg/ml) was added and incubated at 37 °C for 3 h, after which the medium was removed and 100 µl of DMSO was added, and shaken for about 5 min on an orbital shaker. Formazan crystals formed were allowed to dissolve in DMSO. The absorbance of DMSO solubilized formazan was read at 540 nm (Shi et al. 2015). IC50 of Catechin was calculated by fitting the data to non-linear regression using GraphPad Prism.

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Cell cycle analysis[2]
Flow cytometric technique was used to evaluate the effect of Catechin on DOX- induced alteration of cell cycle. One million differentiated cells were seeded in flasks for overnight and incubated with Catechin (40 µg/ml) at 37 °C for 2 h, followed by DOX at 1.5 µg/ml for next 24 h. Cells were separated from the flask by trypsinization, washed with PBS with centrifugation. The cell pellets were collected and fixed with 70% ice-cold methanol and stored for 24 h at −20 °C. Then cell pellets were washed with PBS and added isotonic PI solution (25 µM propidium iodide, 0.03% NP-40 and 40 µg/ml RNase A) for staining. The stained cells were analyzed with Accuri C6 flow cytometer for cell cycle study at excitation wavelength 488 nm and emission wavelength 575/40 nm (Reddy et al. 2015; Simon et al. 2016).

In vivo studies[2]
Selection of doses In the preliminary experiments for assessing the procognitive effect of Catechin, the selected doses were 50, 100 and 200 mg/kg for dose–response analysis. In later studies for chemobrain i.e., DOX-induced memory deficit model, the dose of Catechin selected was 100 mg/kg as it showed a promising effect in preliminary studies and moreover, the treatment was on a chronic basis. The dose of DOX selected was 2.5 mg/kg according to the previous studies and standardized laboratory procedures (Steiniger et al. 2004; Swamy et al. 2011; Grandhi et al. 2016).[2]
Preparation and administration of drugs In the preliminary study for assessing the nootropic effect of Catechin using time induced memory deficit model, the doses were prepared at 50, 100, 200 mg/kg in 0.25% w/v sodium carboxy methylcellulose (CMC) and administered orally for 7 days prior to and during the experimental trials. Four experimental groups were used (n = 9 each) for one vehicle (CMC) and three groups of Catechin (three doses).[2]
For inducing neurotoxicity and systemic toxicity, DOX (Adriamycin at 2.5 mg/kg) was administered intraperitoneally in 10 cycles on every 5 days. Three experimental groups (n = 12 each) were used viz., vehicle control (0.25% w/v CMC), DOX alone and Catechin (100 mg/kg in 0.25% CMC p.o.). Catechin was administered orally for 57 days including one-week treatment prior to the first cycle of DOX. Following the last cycle of DOX on day 57, i.e., on day 58, the behavioral study was conducted. All treatments, as well as the behavioral analysis, were carried out between 9 a.m. to 4 p.m. Body weight of the animals was taken once in 3 days throughout the study. During the experimental trials, the oral treatment was given 1 h before the familiarization trial.

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Time-induced natural memory deficits model[2]
Episodic memory deficits can be induced in rats naturally by increasing the time delay between familiarization and choice trials. Hence the initial experiment was conducted to assess the effect of Catechin on time -induced memory deficits by using an ITI of 24 h. In this test, rats were habituated to the arenas on day 1 and were subjected to familiarization trial on day 2. Then after an ITI of 24 h, i.e., on day 3, animals were subjected to recognition trial with one novel object replacing the familiar object. Four experimental groups were used. Rats were treated with either Catechin (50, 100 and 200 mg/kg, p.o.) or CMC (10 ml/kg, p.o.) for 7 days before the trial initiation. During the experimental trials, treatment was given 1 h before the actual trial in familiarization and choice trials

[1]. Potential cancer-chemopreventive activities of wine stilbenoids and flavans extracted from grape (Vitis vinifera) cell cultures. Nutr Cancer. 2001;40(2):173-9.

[2]. Catechin ameliorates doxorubicin-induced neuronal cytotoxicity in in vitro and episodic memory deficit in in vivo in Wistar rats. Cytotechnology. 2018 Feb;70(1):245-259.

[3]. Alshatwi AA. Catechin hydrate suppresses MCF-7 proliferation through TP53/Caspase-mediated apoptosis. J Exp Clin Cancer Res. 2010 Dec 17;29:167.

(+)-catechin monohydrate is the monohydrate of (+)-catechin. It has a role as a geroprotector. It contains a (+)-catechin.

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(+)-catechin is the (+)-enantiomer of catechin and a polyphenolic antioxidant plant metabolite. It has a role as an antioxidant and a plant metabolite. It is an enantiomer of a (-)-catechin.
An antioxidant flavonoid, occurring especially in woody plants as both (+)-catechin and (-)-epicatechin (cis) forms.
Cianidanol has been reported in Camellia sinensis, Paeonia obovata, and other organisms with data available.
Catechin is a metabolite found in or produced by Saccharomyces cerevisiae.
An antioxidant flavonoid, occurring especially in woody plants as both (+)-catechin and (-)-epicatechin (cis) forms.
See also: Gallocatechin (has subclass); Crofelemer (monomer of); Bilberry (part of) ...

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Moderate consumption of wine is associated with a reduced risk of cancer. Grape plant cell cultures were used to purify 12 phenols: the stilbenoids trans-astringin, trans-piceid (2), trans-resveratroloside, trans-resveratrol, trans-piceatannol, cis-resveratroloside, cis-piceid, and cis-resveratrol; the flavans (+)-catechin, (-)-epicatechin, and epicatechin 3-O-gallate; and the flavan dimer procyanidin B2 3'-O-gallate. These compounds were evaluated for potential to inhibit cyclooxygenases and preneoplastic lesion formation in carcinogen-treated mouse mammary glands in organ culture. At 10 micrograms/ml, trans-astringin and trans-piceatannol inhibited development of 7,12-dimethylbenz[a]anthracene-induced preneoplastic lesions in mouse mammary glands with 68.8% and 76.9% inhibition, respectively, compared with untreated glands. The latter compound was the most potent of the 12 compounds tested in this assay, with the exception of trans-resveratrol (87.5% inhibition). In the cyclooxygenase (COX)-1 assay, trans isomers of the stilbenoids appear to be more active than cis isomers: trans-resveratrol [50% inhibitory concentration (IC50) = 14.9 microM, 96%] vs. cis-resveratrol (IC50 = 55.4 microM). In the COX-2 assay, among the compounds tested, only trans- and cis-resveratrol exhibited significant inhibitory activity (IC50 = 32.2 and 50.2 microM, respectively). This is the first report showing the potential cancer-chemopreventive activity of trans-astringin, a plant stilbenoid recently found in wine. trans-Astringin and its aglycone trans-piceatannol were active in the mouse mammary gland organ culture assay but did not exhibit activity in COX-1 and COX-2 assays. trans-Resveratrol was active in all three of the bioassays used in this investigation. These findings suggest that trans-astringin and trans-piceatannol may function as potential cancer-chemopreventive agents by a mechanism different from that of trans-resveratrol.[1]

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Cognitive dysfunction by chemotherapy compromises the quality of life in cancer patients. Tea polyphenols are known chemopreventive agents. The present study was designed to evaluate the neuroprotective potential of (+) catechin hydrate (catechin), a tea polyphenol, in IMR-32 neuroblastoma cells in vitro and alleviation of episodic memory deficit in Wistar rats in vivo against a widely used chemotherapeutic agent, Doxorubicin (DOX). In vitro, neuroprotective studies were assessed in undifferentiated IMR-32 cells using percentage viability and in differentiated cells by neurite length. These studies showed catechin increased percentage viability of undifferentiated IMR-32 cells. Catechin pretreatment also showed an increase in neurite length of differentiated cells. In vivo neuroprotection of catechin was evaluated using novel object recognition task in time-induced memory deficit model at 50, 100 and 200 mg/kg dose and DOX-induced memory deficit models at 100 mg/kg dose. The latter model was developed by injection of DOX (2.5 mg/kg, i.p.) in 10 cycles over 50 days in Wistar rats. Catechin showed a significant reversal of time-induced memory deficit in a dose-dependent manner and prevention of DOX-induced memory deficit at 100 mg/kg. In addition, catechin treatment showed a significant decrease in oxidative stress, acetylcholine esterase and neuroinflammation in the hippocampus and cerebral cortex in DOX-induced toxicity model. Hence, catechin may be a potential adjuvant therapy for the amelioration of DOX-induced cognitive impairment which may improve the quality of life of cancer survivors. This improvement might be due to the elevation of antioxidant defense, prevention of neuroinflammation and inhibition of acetylcholine esterase enzyme.[2]

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Catechin hydrate (CH), a strong antioxidant that scavenges radicals, is a phenolic compound that is extracted from plants and is present in natural food and drinks, such as green tea and red wine. CH possesses anticancer potential. The mechanism of action of many anticancer drugs is based on their ability to induce apoptosis. In this study, I sought to characterize the downstream apoptotic genes targeted by CH in MCF-7 human breast cancer cells. CH effectively kills MCF-7 cells through induction of apoptosis. Apoptosis was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and real-time PCR assays. Cells were exposed to 150 μg/ml CH and 300 μg/mL CH for 24 hours, which resulted in 40.7% and 41.16% apoptotic cells, respectively. Moreover, a 48-hour exposure to 150 μg/ml CH and 300 μg/ml CH resulted in 43.73% and 52.95% apoptotic cells, respectively. Interestingly, after 72 hours of exposure to both concentrations of CH, almost 100% of cells lost their integrity. These results were further confirmed by the increased expression of caspase-3,-8, and -9 and TP53 in a time-dependent and dose-dependent manner, as determined by real-time quantitative PCR. In summary, the induction of apoptosis by CH is affected by its ability to increase the expression of pro-apoptotic genes such as caspase-3, -8, and -9 and TP53.[3]

C15H14O6

290.2681

308.089

C, 58.44; H, 5.23; O, 36.33

225937-10-0

(±)-Catechin;7295-85-4;Catechin;154-23-4

107957

Off-white to yellow solid powder

630.4ºC at760mmHg

175-177ºC

335ºC

9.29E-17mmHg at 25°C

1.481

6

7

1

22

364

2

C1[C@@H]([C@H](OC2=CC(=CC(=C21)O)O)C3=CC(=C(C=C3)O)O)O.O

OFUMQWOJBVNKLR-NQQJLSKUSA-N

InChI=1S/C15H14O6.H2O/c16-8-4-11(18)9-6-13(20)15(21-14(9)5-8)7-1-2-10(17)12(19)3-7;/h1-5,13,15-20H,6H2;1H2/t13-,15+;/m0./s1

(2R,3S)-2-(3,4-dihydroxyphenyl)-3,4-dihydro-2H-chromene-3,5,7-triol;hydrate

(+)-Catechin Hydrate; 225937-10-0; Catechin hydrate; 88191-48-4; (+)-catechin monohydrate; (2R,3S)-2-(3,4-Dihydroxyphenyl)chroman-3,5,7-triol hydrate; MFCD00149354; (+)-Cyanidol-3;

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 : ~50 mg/mL

Solubility in Formulation 1: ≥ 2.5 mg/mL (Infinity mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (Infinity mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (Infinity mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)