Theacrine

Theacrine, a Potent Antidepressant Purine Alkaloid from a Special Chinese Tea, Promotes Adult Hippocampal Neurogenesis in Stressed Mice.[Pubmed:34060828]

J Agric Food Chem. 2021 Jun 30;69(25):7016-7027.

Daily intake of tea has been known to relate to a low risk of depression. In this study, we report that a special variety of tea in China, Camellia assamica var. kucha (kucha), possesses antidepressant effects but with less adverse effects as compared to traditional tea Camellia sinensis. This action of kucha is related to its high amount of Theacrine, a purine alkaloid structurally similar to caffeine. We investigated the antidepressant-like effects and mechanisms of Theacrine in chronic water immersion restraint stress and chronic unpredictable mild stress mice models. PC12 cells and primary hippocampal neural stem cells were treated with stress hormone corticosterone (CORT) to reveal the potential antidepression mechanism of Theacrine from the perspective of adult hippocampus neurogenesis. Results of behavioral and neurotransmitter analysis showed that intragastric administration of Theacrine significantly counteracted chronic stress-induced depression-like disorders and abnormal 5-hydroxytryptamine (5-HT) metabolism with less central excitability. Further investigation from both in vivo and in vitro experiments indicated that the antidepressant mechanism of Theacrine was associated with promoting adult hippocampal neurogenesis, via the modulation of the phosphodiesterase-4 (PDE4)/cyclic adenosine monophosphate (cAMP)/cAMP response-element binding (CREB)/brain-derived neurotrophic factor (BDNF)/tropomyosin-related kinase B (TrkB) pathway. Collectively, our findings could promote the prevalence of kucha as a common beverage with uses for health care and contribute to the development of Theacrine as a potential novel antidepressant medicine.

Theacrine From Camellia kucha and Its Health Beneficial Effects.[Pubmed:33392238]

Front Nutr. 2020 Dec 17;7:596823.

Theacrine, i.e., 1,3,7,9-tetramethyluric acid, is one of the major purine alkaloids found in leaf of a wild tea plant species Camellia kucha Hung T. Chang. Theacrine has been attracted great attentions academically owing to its diverse health benefits. Present review examines the advances in the research on the health beneficial effects of Theacrine, including antioxidant effect, anti-inflammatory effect, locomotor activation and reducing fatigue effects, improving cognitive effect, hypnotic effect, ameliorating lipid metabolism and inhibiting breast cancer cell metastasis effect. The inconsistent results in this research field and further expectations were also discussed.

Identification of key metabolites based on non-targeted metabolomics and chemometrics analyses provides insights into bitterness in Kucha [Camellia kucha (Chang et Wang) Chang].[Pubmed:33288175]

Food Res Int. 2020 Dec;138(Pt B):109789.

Camellia kucha (Chang et Wang) Chang is a special tea in China, which is extremely bitter but beneficial for human health. However, there are no systematic studies on Kucha metabolites, especially those associated with bitterness. In this study, a non-targeted metabolomics approach based on UHPLC-LTQ-Orbitrap-MS was applied to comprehensively profile the characteristic metabolites of two Kucha cultivars by comparison with three common tea cultivars. A total of 90 differential metabolites were identified. Among them, eight key metabolites (Theacrine, 2,4-dimethyl-1H-indole, EGCG, dihydrokaempferol, panasenoside, 3-cresotinic acid, 3-methylglutaconic acid, and L-histidine) were more abundant in Kucha than in the controls, most of which were positively correlated with bitterness. Furthermore, quantitative analysis of some important catechins and alkaloids by HPLC implied absolutely higher concentrations of EGCG and Theacrine in Kucha, which was similar to the metabolomics results. These results will be contribute to future research on the bitter and nutritional properties of Kucha.

A Theacrine-Based Supplement Increases Cellular NAD(+) Levels and Affects Biomarkers Related to Sirtuin Activity in C2C12 Muscle Cells In Vitro.[Pubmed:33287129]

Nutrients. 2020 Dec 3;12(12). pii: nu12123727.

There is evidence in rodents to suggest that Theacrine-based supplements modulate tissue sirtuin activity as well as other biological processes associated with aging. Herein, we examined if a Theacrine-based supplement (termed NAD3) altered sirtuin activity in vitro while also affecting markers of mitochondrial biogenesis. The murine C2C12 myoblast cell line was used for experimentation. Following 7 days of differentiation, myotubes were treated with 0.45 mg/mL of NAD3 (containing ~2 mM Theacrine) for 3 and 24 h (n = 6 treatment wells per time point). Relative to control (CTL)-treated cells, NAD3 treatments increased (p < 0.05) Sirt1 mRNA levels at 3 h, as well as global sirtuin activity at 3 and 24 h. Follow-up experiments comparing 24 h NAD3 or CTL treatments indicated that NAD3 increased nicotinamide phosphoribosyltransferase (NAMPT) and SIRT1 protein levels (p < 0.05). Cellular nicotinamide adenine dinucleotide (NAD(+)) levels were also elevated nearly two-fold after 24 h of NAD3 versus CTL treatments (p < 0.001). Markers of mitochondrial biogenesis were minimally affected. Although these data are limited to select biomarkers in vitro, these preliminary findings suggest that a Theacrine-based supplement can modulate select biomarkers related to NAD(+) biogenesis and sirtuin activity. However, these changes did not drive increases in mitochondrial biogenesis. While promising, these data are limited to a rodent cell line and human muscle biopsy studies are needed to validate and elucidate the significance of these findings.

Development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for characterizing caffeine, methylliberine, and theacrine pharmacokinetics in humans.[Pubmed:32829142]

J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Oct 15;1155:122278.

Coffea liberica possesses stimulant properties without accumulating the methylxanthine caffeine. The basis for this peculiar observation is that methylurates (e.g., Theacrine and methylliberine) have replaced caffeine. The stimulant properties of methylurates, alone and in combination with caffeine, have recently been investigated. However, human pharmacokinetics and LC-MS/MS methods for simultaneous measurement of methylxanthines and methylurates are lacking. To address this deficiency, we conducted a pharmacokinetic study in which subjects (n = 12) were orally administered caffeine (150 mg), methylliberine (Dynamine, 100 mg), and Theacrine (TeaCrine(R), 50 mg) followed by blood sampling over 24 h. Liquid-liquid extraction of plasma samples containing purine alkaloids and internal standard ((13)C-Caffeine) were analyzed using a C18 reversed-phase column and gradient elution (acetonitrile and water, both containing 0.1% formic acid). A Waters Xevo TQ-S tandem mass spectrometer (positive mode) was used to detect caffeine, methylliberine, Theacrine, and IS transitions of m/z 195.11 --> 138.01, 225.12 --> 168.02, 225.12 --> 167.95, and 198.1 --> 140.07, respectively. The method was validated for precision, accuracy, selectivity, and linearity and was successfully applied to characterize the oral pharmacokinetics of caffeine, methylliberine, and Theacrine in human plasma. Successful development and application of LC-MS/MS-based methods such as ours for the simultaneous measurement of methylxanthines and methylurates are essential for the characterization of potential pharmacokinetic and pharmacodynamic interactions.

Theacrine and strictinin, two major ingredients for the anti-influenza activity of Yunnan Kucha tea.[Pubmed:32730889]

J Ethnopharmacol. 2020 Nov 15;262:113190.

ETHNOPHARMACOLOGICAL RELEVANCE: Kucha tea plant (Camellia assamica var. kucha Chang et Wang) is regarded as a mutant variety of wild Pu'er tea plant found in few mountain areas of Yunnan, China. Its fresh young leaves and shoots are picked by the indigenous aborigines in these local areas to prepare an herbal tea for the treatment of common cold empirically. MATERIALS AND METHODS: Two extra compounds of relative abundance were detected in Kucha tea in comparison with Pu'er tea, and their chemical structures were identified as chlorogenic acid and Theacrine. These two compounds as well as two major compounds, strictinin and caffeine, in Kucha tea were evaluated for their cytotoxicity and inhibitory effects on human influenza virus A/Puerto Rico/8/34 by analyzing viral protein expression and progeny production. RESULTS: No or low cytotoxicity was detected for the four Kucha compounds when their concentrations were below 100 muM. Expression of viral NS1 protein was significantly inhibited by chlorogenic acid, Theacrine or strictinin, but not caffeine at a concentration of 100 muM. The relative inhibitory potency was detected as chlorogenic acid < Theacrine < strictinin, and both Theacrine and strictinin displayed significant inhibition at a concentration of 50 muM. According to a plaque assay, viral progeny production was significantly reduced by Theacrine or strictinin, but not by chlorogenic acid or caffeine under the same concentration of 100 muM. CONCLUSION: It is suggested that Theacrine and strictinin are two major ingredients responsible for the anti-influenza activity of Yunnan Kucha tea traditionally used for the treatment of common cold.

Theacrine, a purine alkaloid from kucha, protects against Parkinson's disease through SIRT3 activation.[Pubmed:32707370]

Phytomedicine. 2020 Oct;77:153281.

BACKGROUND: Oxidative damage of dopaminergic neurons is the fundamental causes of Parkinson's disease (PD) that has no standard cure at present. Theacrine, a purine alkaloid from Chinese tea Kucha, has been speculated to benefit the neurodegeneration in PD, through similar actions to its chemical analogue caffeine, albeit excluding side effects. Theacrine has nowadays gained a lot of interest for its multiple benefits, while the investigations are weak and insufficient. HYPOTHESIS/PURPOSE: It is well-known that tea has a wide range of functions, especially in the prevention and treatment of neurodegenerative diseases. Theacrine is an active monomer compound in Camellia assamica var. kucha Hung T. Chang & H.S.Wang (Kucha), which appears to be effective and safe in PD therapy. The aim of this study is to examine its actions in diverse PD models and explore the mechanisms. STUDY DESIGN: For determination of Theacrine's effects, we employed diverse oxidative damage-associated PD models, including 6-OHDA-treated rats, MPTP-treated mice/zebrafish and MPP(+)-treated SH-SY5Y cells, and using caffeine, selegiline and depranyl as positve control. For investigation and verification of the mechanisms, we utilized approaches testing mitochondrial function-related parameters and enzyme activity as well as applied gene knockdown and overexpression. METHODS: We employed behavioral tests including spontaneous activity, pole, swimming, rotarod and gait, immunohistochemistry, HPLC, flow cytometry, immunohistochemistry, Western blot, gene knockdown by siRNA and overexpression by plasmid in this study. RESULTS: Theacrine is demonstrated to retrieve the loss of dopaminergic neurons and the damages of behavioral performance in multiple animal models of PD (6-OHDA-treated rats and in MPTP-treated mice and zebrafish). The followed data of MPP(+)-treated SH-SY5Y cells indicate that Theacrine relieves apoptosis resulted from oxidative damage and mitochondrial dysfunction. Further investigations illustrate that Theacrine activates SIRT3 directly. It is of advantage to prevent apoptosis through SIRT3-mediated SOD2 deacetylation that reduces ROS accumulation and restores mitochondrial function. This concept is elaborated by 3TYP that inhibits SIRT3 enzyme activity and knockdown/overexpression of SIRT3 gene, demonstrating a crucial role of SIRT3 in Theacrine-benefited dopaminergic neurons. CONCLUSION: Theacrine prevents apoptosis of dopaminergic neurons through directly activating SIRT3 which deacetylating SOD2 and restoring mitochondrial functions.

Baiyacha, a wild tea plant naturally occurring high contents of theacrine and 3''-methyl-epigallocatechin gallate from Fujian, China.[Pubmed:32546720]

Sci Rep. 2020 Jun 16;10(1):9715.

Baiyacha (BYC) is a kind of wild tea plant growing and utilizing in the remote mountain area of Fujian province, Southeastern China. However, scientific studies on this plant remain limited. Our results showed that BYC exhibits the typical morphological characteristics of Camellia gymnogyna Chang, a closely related species of C. sinensis (L.) O. Kuntze, which was not found in Fujian before. Chemical profiling revealed that parts of BYC plants are rich in purine alkaloids and catechins, especially featuring high levels of Theacrine and 3''-methyl-epigallocatechin gallate (EGCG3''Me), chemical compounds with multiple biological activities that are rarely observed in regular tea plants. The contents of EGCG3''Me and Theacrine in BYC both increased with the leaf maturity of tea shoots, whereas the caffeine content decreased significantly. The obtained results provide abundant information about the morphology and chemical compounds of BYC and may be used for tea production, breeding, and scientific research in the future.

Novel insight into theacrine metabolism revealed by transcriptome analysis in bitter tea (Kucha, Camellia sinensis).[Pubmed:32286351]

Sci Rep. 2020 Apr 14;10(1):6286.

Kucha (Camellia sinensis) is a kind of unique wild tea resources in southwest China, containing sizeable amounts of Theacrine (1,3,7,9-tetramethyluric acid) and having a special bitter taste both in fresh leaves and made tea. Theacrine has good healthy function locally. But the molecular mechanism of Theacrine metabolism in Kucha was still unclear. In order to illuminate the biosynthesis and catabolism of Theacrine in Kucha plants, three tea cultivars, C. sinensis 'Shangyou Zhongye' (SY) with low-Theacrine, 'Niedu Kucha 2' (ND2) with middle-Theacrine and, 'Niedu Kucha 3' (ND3) with high-Theacrine, were used for our research. Purine alkaloid analysis and transcriptome of those samples were performed by High Performance Liquid Chromatography (HPLC) and RNA-Seq, respectively. The related gene expression levels of purine alkaloid were correlated with the content of purine alkaloid, and the results of quantitative real-time (qRT) PCR were also confirmed the reliability of transcriptome. Based on the data, we found that Theacrine biosynthesis is a relatively complex process, N-methyltransferase (NMT) encoded by TEA024443 may catalyze the methylation at 9-N position in Kucha plant. Our finding will assist to reveal the molecular mechanism of Theacrine biosynthesis, and be applied to selection and breeding of Kucha tea cultivars in the future.

Identification and characterization of N9-methyltransferase involved in converting caffeine into non-stimulatory theacrine in tea.[Pubmed:32193380]

Nat Commun. 2020 Mar 19;11(1):1473.

Caffeine is a major component of xanthine alkaloids and commonly consumed in many popular beverages. Due to its occasional side effects, reduction of caffeine in a natural way is of great importance and economic significance. Recent studies reveal that caffeine can be converted into non-stimulatory Theacrine in the rare tea plant Camellia assamica var. kucha (Kucha), which involves oxidation at the C8 and methylation at the N9 positions of caffeine. However, the underlying molecular mechanism remains unclear. Here, we identify the Theacrine synthase CkTcS from Kucha, which possesses novel N9-methyltransferase activity using 1,3,7-trimethyluric acid but not caffeine as a substrate, confirming that C8 oxidation takes place prior to N9-methylation. The crystal structure of the CkTcS complex reveals the key residues that are required for the N9-methylation, providing insights into how caffeine N-methyltransferases in tea plants have evolved to catalyze regioselective N-methylation through fine tuning of their active sites. These results may guide the future development of decaffeinated drinks.

Safety of Short-Term Supplementation with Methylliberine (Dynamine((R))) Alone and in Combination with TeaCrine((R)) in Young Adults.[Pubmed:32121218]

Nutrients. 2020 Feb 28;12(3). pii: nu12030654.

Methylliberine (Dynamine((R)); DYM) and Theacrine (Teacrine((R)); TCR) are purine alkaloids purported to have similar neuro-energetic effects as caffeine. There are no published human safety data on DYM, and research on TCR is limited. The purpose of this study was to examine the effect of four weeks of DYM supplementation with and without TCR on cardiovascular function and blood biomarkers. One-hundred twenty-five men and women (mean age 23.0 yrs, height 169.7 cm, body mass 72.1 kg; n = 25/group) were randomly assigned to one of five groups: low-dose DYM (100 mg), high-dose DYM (150 mg), low-dose DYM with TCR (100 mg + 50 mg), high-dose DYM with TCR (150 mg + 25 mg) , and placebo. Regardless of group and sex, significant main effects for time were noted for heart rate, systolic blood pressure, and QTc (p < 0.001), high-density lipoproteins (p = 0.002), mean corpuscular hemoglobin (p = 0.018), basophils (p = 0.006), absolute eosinophils (p = 0.010), creatinine (p = 0.004), estimated glomerular filtration rate (p = 0.037), chloride (p = 0.030), carbon dioxide (p = 0.023), bilirubin (p = 0.027), and alanine aminotransferase (p = 0.043), among others. While small changes were found in some cardiovascular and blood biomarkers, no clinically significant changes occurred. This suggests that DYM alone or in combination with TCR consumed at the dosages used in this study does not appear to negatively affect markers of health over four weeks of continuous use.

Theacrine alleviates chronic inflammation by enhancing TGF-beta-mediated shifts via TGF-beta/SMAD pathway in Freund's incomplete adjuvant-induced rats.[Pubmed:31791581]

Biochem Biophys Res Commun. 2020 Feb 12;522(3):743-748.

Rheumatoid arthritis is a chronic and systemic autoimmune disease, which affects approximately 1% of the adult population worldwide. The present study investigated the therapeutic effect of Theacrine (TC) on arthritis and its mechanisms in Freund's incomplete adjuvant (FIA)-induced SD rats. Rats were randomly divided into 5 groups: i) healthy control; ii) model; iii) positive control with methotrexate (MTX); iv) treatment with 12.5 mg/kg TC; and v) treatment with 25.0 mg/kg TC. The apparent scores, including changes in body weights, degree of paw swelling and arthritis indicators, were analyzed to evaluate the anti-chronic inflammatory effect of TC. The levels of interleukin (IL)-6 and transforming growth factor-beta (TGF-beta) in serum were measured by enzyme-linked immunosorbent assay. The protein and RNA expression levels of the critical factors in rats were measured to elucidate the mechanisms responsible for chronic inflammation and to verify molecular indexes of chronic inflammatory conditions. TC notably suppressed the severity of FIA-induced rat by attenuating the apparent scores, animal weight and inflammatory indexes in the 25 mg/kg TC group compared with the FIA rat model. Furthermore, TC significantly decreased the levels of IL-6 and increased the levels of TGF-beta. Histopathological examinations indicated that TC rescued the synovial hyperplasia and inflammatory cell infiltration in joint tissues. In addition, TC enhanced TGF-beta-mediated shifts in inflammatory marker expression in joint tissue. Overall, the present study demonstrated that TC exerted a superior anti-arthritic effect via the suppression of IL-6 and the activation of TGF-beta by the TGF-beta/SMAD pathway.

SIRT3 retards intervertebral disc degeneration by anti-oxidative stress by activating the SIRT3/FOXO3/SOD2 signaling pathway.[Pubmed:31773668]

Eur Rev Med Pharmacol Sci. 2019 Nov;23(21):9180-9188.

OBJECTIVE: The objective of this paper is to determine whether SIRT3 could retard intervertebral disc degeneration and study the mechanism. MATERIALS AND METHODS: We chose the 3-month mice to establish intervertebral disc degeneration model and study the effect of SIRT3 on the intervertebral disc by Western blotting, quantitative Real Time-Polymerase Chain Reaction (qRT-PCR), immunohistochemistry. Mouse nucleus pulposus cells were cultured to study the exact mechanism. RESULTS: The expression of SIRT3 was decreased in degenerated human nucleus pulposus. Intervertebral discs of mice treated with Theacrine expressed more collagen II and less collagen X. In addition, nucleus pulposus cells stimulated with interleukin-1beta (IL-1beta) expressed less SIRT3 than that in the control group and nucleus pulposus cells with SIRT3 overexpress vectors expressed more collagen II FOXO3a and superoxide dismutase 2 (SOD2), indicating that SIRT3 could improve the intervertebral disc degeneration by anti-oxidative stress. CONCLUSIONS: SIRT3 is a protective factor for intervertebral discs and can reduce oxidative stress in the intervertebral disc.

Purification and characterization of theobromine synthase in a Theobromine-Enriched wild tea plant (Camellia gymnogyna Chang) from Dayao Mountain, China.[Pubmed:31753680]

Food Chem. 2020 May 1;311:125875.

Camellia gymnogyna Chang (CgC), a wild tea plant, was discovered on Dayao Mountain, China. However, research regarding this tea plant is limited. Our study found that CgC contains theobromine, caffeine, and Theacrine, among which theobromine content was the highest (14.37-39.72mg/g). In addition, theobromine synthase (TS) was partially purified from CgC leaves, up to 35.87-fold, with consecutive chromatography, and its molecular weight was found to be approximately 62kDa. The optimum reaction time, pH, and temperature for theobromine synthase from 7-methylxanthine was found to be 6h, 4, and 45 degrees C, respectively. TS expression at both mRNA and protein stages was higher in the first than in the fourth leaf (P<0.05). Subcellular localization of TS indicated that it was localized in the nucleus. These results indicate that CgC can be of scientific value and could lead to efficient utilization of this rare wild tea germplasm.

The effects of a caffeine-like supplement, TeaCrine(R), on muscular strength, endurance and power performance in resistance-trained men.[Pubmed:31660991]

J Int Soc Sports Nutr. 2019 Oct 28;16(1):47.

BACKGROUND: TeaCrine(R) is the synthetic version to naturally occurring Theacrine (1, 3, 7, 9-tetramethyluric acid) found in the leaves of Camellia kucha tea plants. A few studies have examined the effects of TeaCrine(R) on cognitive perception, but no research exists examining its effects on resistance exercise performance. The purpose of this study was to determine the efficacy of TeaCrine(R), a caffeine-like compound, on maximal muscular strength, endurance, and power performance in resistance-trained men. METHODS: Twelve resistance-trained men participated in a randomized, double-blind, cross-over designed study. Each participant performed one-repetition maximum (1RM) bench press, 1RM squat, bench press repetitions to failure (RTF) at 70% 1RM, squat RTF at 70% 1RM, and 2-km rowing time trial 90 min after consumption of: (1) Caffeine 300 mg (CAFF300); (2) TeaCrine(R) 300 mg (TEA300); (3) TeaCrine(R) + Caffeine (COMBO; 150 mg/150 mg); (4) Placebo 300 mg (PLA). Power and velocity were measured using a TENDO Power Analyzer. Visual analogue scales for energy, focus, motivation to exercise, and fatigue were administered at baseline and 90 min post-treatment ingestion (pre-workout). Rating of perceived exertion was assessed after bench press RTF and squat RTF. RESULTS: There were no differences between groups for 1RM, RTF, and power in the bench press and squat exercises. Only CAFF300 resulted in significant increases in perceived energy and motivation to exercise vs. TEA300 and PLA (Energy: + 9.8%, 95% confidence interval [3.3-16.4%], p < 0.01; + 15.3%, 95% CI [2.2-28.5%], p < 0.02; Motivation to exercise: + 8.9%, 95% CI [0.2-17.6%], p = 0.04, + 14.8%, 95% CI [4.7-24.8%], p < 0.01, respectively) and increased focus (+ 9.6%, 95% CI [2.1-17.1%], p = 0.01) vs. TEA300, but there were no significant differences between CAFF300 and COMBO (Energy + 3.9% [- 6.9-14.7%], Focus + 2.5% [- 6.3-11.3%], Motivation to exercise + 0.5% [- 11.6-12.6%]; p > 0.05). CONCLUSION: Neither TEA300, CAFF300, COMBO, or PLA (when consumed 90 min pre-exercise) improved muscular strength, power, or endurance performance in resistance-trained men. Only CAFF300 improved measures of focus, energy, and motivation to exercise.