Effect of Cellgevity® Supplement on Selected Rat Liver Cytochrome P450 Enzyme Activity and Pharmacokinetic Parameters of Carbamazepine

Effect of Cellgevity® Supplement on Selected Rat Liver Cytochrome P450 Enzyme Activity and Pharmacokinetic Parameters of Carbamazepine | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research article Effect of Cellgevity® Supplement on Selected Rat Liver Cytochrome P450 Enzyme Activity and Pharmacokinetic Parameters of Carbamazepine Seth Kwabena Amponsah, Benoit Banga Nguessan, Martin Akandawen, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.2.19774/v2 This work is licensed under a CC BY 4.0 License Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Abstract Background : There is considerable evidence that many patients concurrently administer dietary supplements with conventional drugs, creating a risk for potential drug-supplement interaction. The aim of this study was to determine the effect of Cellgevity® supplement on selected rat liver cytochrome P450 (CYP) enzymes. Also, we sought to deternine the effect of Cellgevity® on the pharmacokinetics of carbamazepine, a CYP3A4 substrate. Methods: Male Sprague-Dawley (SD) rats were randomly put into 5 groups and administered ( per os ) either distilled water (negative control), Cellgevity® (3 separate doses) or phenobarbital (positive control). Modulation of liver CYP enzyme activity was evaluated after 30 days of treatment, using probe substrates, spectroscopic and high-performance liquid chromatographic methods. In the pharmacokinetic study, 12 SD rats were divided into 2 groups administered ( per os ) carbamazepine plus Cellgevity®, or carbamazepine plus normal saline, both over a period of 14 days. Blood samples from rats in the same group were collected after treatment. Serum samples were prepared and pooled together at each specific sampling time point. Levels of carbamazepine were determined using a fluorescence polarization immunoassay. Results: Activities of rat liver CYP1A1/2, CYP2C9 and CYP2D6 were significantly increased by Cellgevity® after 30-day treament. Pharmacokinetic parameters for rats administered carbamazepine with Cellgevity® vis-a-vis carbamazepine with normal saline were as follows: C max ; 20 μmol/L vs 11 μmol/L, AUC 0→24 ; 347 μmol.h/L vs 170 μmol.h/L, K e ; 0.28 h -1 vs 0.41 h -1 , and t 1/2 ; 2.3 h vs 1.7 h, respectively. Conclusions: Cellgevity® increased the activity of rat CYP1A1/2, CYP2C9 and CYP2D6 enzymes, and altered the pharmacokinetics of carbamazepine in rats. Clinical Pharmacology Carbamazepine Cellgevity® Cytochrome P450 Interaction Pharmacokinetics Supplement Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Background Dietary supplements may be vitamins, minerals or herbal products that are known to improve the well-being of humans (1). This clearly denotes the use of these supplements as an addition to dietary requirements that may not be met by daily meals. However, under no circumstance should dietary supplements be used as replacement for daily meals. The United States Food and Drugs Administration also prohibits the indication of dietary supplements as treatment for diseases or any indication that connotes dietary supplements as primary therapeutic agents (1). With increase in the incidence of non-communicable diseases (NCDs) such as diabetes, cardiomyopathies, cancers and epilepsy, which are often associated with oxidative stress, people resort to the use of dietary supplements (known antioxidants) to prevent these diseases (2). Interestingly, some individuals who use dietary supplements have the notion that these agents may enhance the effects of conventional drugs (3). This assertion may be false. It is noteworthy, however, that synergy between some dietary supplements and conventional drugs have been reported (4). Currently available on the market are a number of dietary supplements known to replenish levels of reduced glutathione, a free radical scavenger, in cells of humans. One such supplement is Cellgevity® which contains a glutathione precursor molecule, riboceine (D-ribose-L-cysteine). Riboceine is known to deliver cysteine into cells and enhance reduced glutathione level in the body (5, 6). The other constituents of Cellgevity® are broccoli seed extract, turmeric root extract, resveratrol, grape seed extract, quercetin, curcumin, milk thistle, vitamin C, selenomethionine, cordyceps, black pepper and aloe extract. Some of these constituents of Cellgevity® are known inducers and/or inhibitors of cytochrome P450 (CYP) enzymes (7, 8). Cellgevity® is marketed and distributed by Max International, which has branches in 14 countries (United States, Nigeria, Cote d’Ivoire, New Zealand, Singapore, Costa Rica, Columbia, Philippines, El Salvador, Malaysia, Guatemala, Ghana and Hong Kong). Cellgevity® has gained popularity in these countries due to the fact that patronizers know that the supplement has a high antioxidant potential (9). Reports suggest that there could be clinically important modulation of CYP enzyme activity by supplements and/or herbal products. This could result in adverse or sub-therapeutic effects of concurrently administered conventional drugs. For example, St. John’s wort was reported to decrease the serum concentration of theophylline (a bronchodilator) as a result of CYP enzyme induction (10). This interaction between St John’s wort and theophylline could lead to sub-therapeutic effect of normal doses of theophylline when there is co-administration. For this reason, patients are often advised not to take theophylline concomitantly with St. John’s wort. Most of the individual constituents of Cellgevity® can modulate xenobiotic metabolizing enzyme activity, and several studies have reported these. However, there is paucity of scientific data on the effect of all these constituents (extracts) combined in Cellgevity® on xenobiotic metabolizing enzymes (CYP enzymes) . In our earlier study, we reported that Cellgevity® at 4 and 8 mg/kg significantly inhibited rat liver CYP2C9, CYP2B1/2B2 and CYP3A4 after a 7-day treatment period (11). In animal models, it is relevant that doses of test agents (drugs/food supplements/herbal products) are at equivalent doses of what pertains in humans. Therefore, as a follow-up to our earlier study, we decided to use animal equivalent doses of Cellgevity® after scaling doses used in humans (12) in the current study. We also decided to administer Cellgevity® over a longer period of time, i.e. 30 days. We have previously reported that Cellgevity ® inhibited rat liver CYP3A4 after a 7-day treatment period (11) . We, therefore, sought to elucidate possible effect of Cellgevity ® on carbamazepine, also known to be extensively metabolized by CYP3A4. This was to aid in assessing potential interaction between these two agents. Carbamazepine is one of the most commonly prescribed drugs in the management of epilepsy. Due to the chronic nature of epilepsy, and the fact that patients have to take carbamazepine for a long time (lifetime in most cases), there is the potential for clinically significant interactions between carbamazepine and co-administered agents like dietary supplements, herbal products and food (13). Therefore, in a parallel experiment, the effect of Cellgevity® on the pharmacokinetics of carbamazepine (a CYP3A4 substrate), was investigated. Methods Animal care and safety This research was approved by the College of Health Sciences Ethical and Protocol Review Committee (Protocol ID: CHS-Et/M.9 – P1.16/2017-2018) of the University of Ghana. All animal procedures used in this study were in accordance with the National Institute of Health Guidelines for the Care and Use of Laboratory Animals. Male Sprague-Dawley (SD) rats, weighing 150-200 g and 6-8 weeks old, were obtained from the Center for Plant Medicine Research, Mampong, Eastern Region, Ghana. The animals were housed in stainless steel cages. Each rat occupied a minimum space of 2 cubic feet (61 cm x 31 cm x 31 cm) with softwood shavings as bedding for their comfort. The SD rats were fed with normal pellet diet (AGRIMAT, Kumasi, Ghana), given water ad libitum , and maintained under standard laboratory conditions (temperature ~25°C, relative humidity 60-70%, and 12 h light-dark cycle). The animals’ feeding area and water troughs were cleaned regularly to prevent contamination. Animals were acclimatized under stated conditions for 7 days before experiment commenced. Hepatic enzyme induction/inhibition studies Animal grouping and treatment administration In determining the effect of Cellgevity® on CYP enzymes after a 30-day treatment period, male SD rats were put into five groups (6 rats in each group). All treatments were per os and for 30 days. Group 1 was administered distilled water, the vehicle used in dissolving Cellgevity® (purchased from Max International, Ghana), and that served as the negative control (N-C) group. Groups 2, 3 and 4 received daily a low dose (L-D) of 38.63 mg/kg Cellgevity®, medium dose (M-D) of 77.25 mg/kg Cellgevity® and high dose (H-D) of 154.50 mg/kg Cellgevity®, respectively. The doses of Cellgevity® administered to SD rats were animal equivalents of what pertains in humans, and calculated as described by Nair AB and Jacob SA (14). The human dose of Cellgevity® is 12.46 mg/kg per os . The SD rats in Group 5 received an oral dose of phenobarbital (Kinapharma, Ghana) 15 mg/kg daily, and that served as the positive control (P-C). After the 30-day treatment period, animals were sacrificed by cervical dislocation. Livers were excised, washed in ice-cold saline solution, weighed, and stored at -80 o C until use. Microsomal Preparation Livers were thawed and homogenized in potassium phosphate buffer (pH 7.4) using a mortar and pestle on ice. Homogenized samples were first centrifuged at 4,000 rpm for 20 min. The supernatant obtained was re-centrifuged (Beckman Avanti J-25, USA) at 25,000 rpm for 2 h. The pellets obtained, microsomes, were collected and stored at -80 o C until use. CYP2C9 ( Diclofenac Hydroxylation) and CYP2D6 ( Dextromethorphan O-demethylation) Assays The assays were performed as previously described (15), with some modification. A volume of 350 µL of 0.1M potassium phosphate buffer (pH 7.4), 50 µL of 1 mM substrate (diclofenac for CYP2C9 assay and dextromethorphan for CYP2D6 – both substrates purchased from Sigma-Aldrich, USA) and 50 µL of 2.5 mg/mL microsome (obtained from rat livers from respective groups) were mixed separately in Eppendorf tubes. The mixtures were pre-incubated at 37 o C for 5 min. A volume of 50 µL of 1 mM nicotinamide adenine dinucleotide phosphate (NADPH) [Sigma-Aldrich, USA] was added, mixed and incubated at 37 o C for 45 min. A 100 µL stopping solution (ZnSO 4 .7H 2 O) was added and the mixture centrifuged at 4000 rpm for 5 min. The supernatants were aliquoted into High-Performance Liquid Chromatography (HPLC, Shimadzu, Japan) vials. Samples were analyzed using HPLC. The chromatographic system consisted of a binary solvent delivery system (LC - 20AB), a degasser (DGU-20A3), an auto-sampler (SIL - 20ACHT), a column temperature controller (CTO - 10AS VP) and a photodiode array detector (SPD - M20A) for CYP2C9 metabolites and fluorescence detector (RF - 10A XL ) for CYP2D6 metabolites. The following chromatographic conditions were used for the analysis of CYP2C9; column, C18 (Shimadzu, Japan), diameter 5 μm, length x width 150 mm × 4.6 mm; flow rate, 1 mL/min; column temperature, 40 o C; injection volume, 20 μL; mobile phase, 20 mM potassium phosphate buffer (pH 7.4)/methanol/acetonitrile (60:22.5:17.5, v/v/v). The same chromatographic conditions were used for the analysis of the CYP2D6, with modification to the mobile phase where 3 solvents were used (acetonitrile/distilled water/triethylamine; 24:75:1, v/v/v). CYP1A1/1A2 - Ethoxyresorufin O-deethylase (EROD), CYP2B1/2B2 - Pentoxyresorufin O-depentylase (PROD) and CYP3A4 - Benzyloxyresorufin O-debenzylase (BROD) Assays The assays were performed as previously described (16, 17), with some modification. In brief, microsomes (CYP enzymes) were tested in a total volume of 100 μL. Aliquots of 70 μL potassium phosphate buffer (pH 7.4) were placed into 96-well black plates. This was followed by addition of 10 μL of 50 μM substrate concentration (resorufin ethyl ether for CYP1A1/2, pentoresorufin for CYP2B1/2 and resorufin benzyl ether for CYP3A4; all substrates purchased from Sigma-Aldrich, USA). The final substrate concentration in 100 μL total reaction volume was 5 μM with 0.25% (v/v) dimethyl sulfoxide (DMSO). It is noteworthy that CYP activities were not expected to be affected at the DMSO concentration used in this experiment (18). Aliquots of 10 μL enzyme (microsome from each rat liver from respective Groups) corresponding to 1 mg/mL protein concentration and the vehicle were added in triplicates. The mixtures were pre-incubated at 37 o C for 5 min. A volume of 10 μL of NADPH was then added to each well and the setup was incubated for 10 min for CYP1A1/2, 20 min for CYP2B1/2 and 30 min for CYP3A4 assays, respectively. Aliquots of 40 μL of stopping solution (20% 0.5 M Tris: 80% acetonitrile) were added to each well and shaken gently. Fluorescence of wells was read at wavelengths of 530 nm excitation and 586 nm emission. Triplicate experiments were performed. The average absorbance of the blank was subtracted from the average absorbance of each sample. Fourteen-day treatment of Cellgevity® on the pharmacokinetics of carbamazepine in rats Animal grouping and treatment administration Twelve male SD rats were obtained for this aspect of the study. The animals were put into 2 groups of 6 (Group 1 and Group 2). All treatments were per os and for 14 days. Group 1 was administered carbamazepine plus saline and Group 2, Cellgevity® plus carbamazepine. A dose of 77.25 mg/kg/day of Cellgevity® plus 80 mg/kg of carbamazepine, both equivalent doses per serving/administration in humans scaled to SD rats (14), were administered to rats in Group 2. Rats in Group 1 received 80 mg/kg/day of carbamazepine plus normal saline (same volume as calculated per rat for the Cellgevity® dose). Blood sample collection After administration of carbamazepine with or without Cellgevity® every 24 h for 14 consecutive days to Groups 1 and 2, tail vein blood samples were taken following the dose administered on the 14 th day. Samples were drawn after 0.5, 1, 4, 12 and 24 h. Blood was collected into microtainer gel tubes and centrifuged at 2000 rpm for 5 min to separate serum, and this was stored at -20ºC until analysis. Assay for carbamazepine in serum Due to low sample volumes from tail veins of SD rats, serum samples from rats in the same group (6 animals) at each time point were pooled together. Such that, for instance, serum samples of Group 1 SD rats at time 0.5, 1, 4, 12 or 24 h, were pooled together to obtain a single sample. Usually, challenges with low sample volume can be circumvented by the approach of sample-pooling (19) . Analysis of carbamazepine in serum was by Fluorescence Polarization Immunoassay (FPIA) [Cobas Integra® 400 Plus, Roche, Philippines]. The lower limit of quantification of serum carbamazepine concentration was 0.8 µmol/L and coefficient of variation was < 5% over the entire calibration range. Statistical analysis CYP activity of treatment groups was expressed as a percentage relative to the negative control group. All values were expressed as mean ± standard deviation. Differences between groups were tested for significance using a One-Way Analysis of Variance (ANOVA). This was followed by post-hoc analysis using Bonferroni’s Multiple Comparison Tests. P -values < 0.05 were considered statistically significant. Non-compartmental pharmacokinetic analysis was used to estimate the various pharmacokinetic parameters of carbamazepine. The maximum serum drug concentration (C max ) and its corresponding time (T max ) were determined by visual inspection of the concentration-time curve. The linear trapezoidal rule was applied in extrapolating area under the concentration-time curves (AUCs) for the two groups. The elimination rate constant (K e ) for both groups were extrapolations (apparent slope) from the last sample time point, i.e. 24 hours. K e for both groups was used to calculate corresponding elimination half-lives (t 1/2 ) . Pharmacokinetic analysis was conducted using GraphPad Prism 7.0. Results Hepatic enzyme induction/inhibition studies All CYPs enzyme activities in the treatment groups were estimated relative to the negative control (N-C) group. CYP2C9 activity in SD rats after 30-day treatment CYP2C9 enzyme activity was higher in the phenobarbital- and Cellgevity®-treated groups in comparison with the N-C group. The phenobarbital- and Cellgevity®-treated groups were significantly different from the N-C group. The increase in rat CYP2C9 enzyme activity by Cellgevity® was dose-dependent. Presentation of the effect of Cellgevity® on rat CYP2C9 enzyme activity in various groups is shown in Figure 1. CYP2D6 activity in SD rats after 30-day treatment CYP2D6 enzyme activity was about 2.5-fold higher in the phenobarbital-treated and about 2-fold higher in the Cellgevity®-treated groups in comparison with the N-C group. The positive control (P-C) and H-D Cellgevity®-treated groups differed significantly (p < 0.01) from the N-C group. The L-D and M-D groups also differed significantly (p < 0.05) in comparison to the N-C group. The increase in rat CYP2D6 enzyme activity by Cellgevity® was not dose-dependent. CYP2D6 enzyme activity of SD rats in various groups after 30-day treatment period is shown in Figure 2. CYP1A1/2 activity in SD rats after 30-day treatment CYP1A1/2 enzyme activity was higher in the phenobarbital- and Cellgevity®-treated groups in comparison with the N-C group. The Cellgevity®-treated L-D and M-D groups showed elevated CYP activity compared to N-C group, but these differences were not statistically significant. However, there was a significant difference between the H-D Cellgevity®-treated group and the N-C group. There was somewhat a dose-dependent increase in rat CYP1A1/2 enzyme activity in the Cellgevity®-treated group. CYP1A1/2 enzyme activity of SD rats in various groups after the 30-day treatment is shown in Figure 3. CYP2B1/2 activity in SD rats after 30-day treatment CYP2B1/2 enzyme activity was elevated in the phenobarbital- and Cellgevity®-treated groups in comparison with the N-C group. The Cellgevity®-treated groups showed elevated levels compared to N-C group, but the differences were not statistically significant. There was also no dose-dependent effect of Cellgevity® on rat CYP2B1/2 enzyme activity. CYP2B1/2 enzyme activity of SD rats in various groups after 30-day treatment is shown in Figure 4. CYP3A4 activity in SD rats after 30-day treatment CYP3A4 enzyme activity was elevated in the phenobarbital- and Cellgevity®-treated groups in comparison with the N-C group. The Cellgevity®-treated groups showed elevated levels compared to the N-C group, but the differences were not statistically significant. There was also no dose-dependent effect of Cellgevity® on rat CYP3A4 enzyme activity. CYP3A4 enzyme activity of SD rats in various groups after 30-day treatment are shown in Figure 5. Overall Effect of Cellgevity® on Rat CYP Enzyme Activity After 30 days, when the Cellgevity®-treated groups were compared to the N-C group, the activities of CYP3A4 and CYP2B1/2 did not differ significantly. However, CYP1A1/2, CYP2C9 and CYP2D6 activities in SD rats treated with Cellgevity® were significantly increased compared to the N-C group. Additionally, the increase in CYP2C9 activity was dose-dependent. The overall effect of Cellgevity® on selected rat CYP enzymes is shown in Table 1. Fourteen-day treatment of Cellgevity® on the pharmacokinetics of carbamazepine in rats Carbamazepine concentration-time curves of rats administered carbamazepine with Cellgevity®, and carbamazepine with normal saline are shown in Figure 6. From the concentration-time curves, rats administered carbamazepine with Cellgevity® had a higher peak concentration at 4 h compared to the rats administered carbamazepine with normal saline. The total carbamazepine exposure (AUC 0→24 ) and peak carbamazepine concentration for rats administered carbamazepine with normal saline were 170 µmol.h/L and 11 µmol/L, respectively, as against 347 µmol.h/L and 20 µmol/L, respectively, for rats administered carbamazepine with Cellgevity®. Elimination rate constants were 0.28 h -1 for SD rats administered carbamazepine with Cellgevity® and 0.41 h -1 for SD rats administered carbamazepine with normal saline. Pharmacokinetic parameters obtained from the concentration-time curves for the two groups are shown in Table 2. Discussion We earlier reported the potential of Cellgevity® to modulate CYP enzymes in rats (11). In that study, 4 mg/kg and 8 mg/kg of Cellgevity® were admistered to SD rats over a period of 7 days. Cellgevity® inhibited rat liver CYP3A4, CYP2C9 and CYP1A2, after the 7-day treatment period (11). In the current study, an equivalent dose of Cellgevity® per serving in humans (12.46 mg/kg) was scaled to SD rats (14), and three doses used over a period of 30 days. Reports suggest that xenobiotics can modulate CYP enzymes depending on dose and treatment duration (12). This study, therefore, sought to investigate the effect of Cellgevity® on rat liver CYP enzymes using 38.63 mg/kg, 77.25 mg/kg and 154.50 mg/kg of Cellgevity® calculated after scaling from humans, and administering Cellgevity® for 30 days. In the present study (after the 30-day treatment period), Cellgevity® significantly increased the activity of rat CYP1A1/2, CYP2C9 and CYP2D6. These results differ from what we earlier reported (11), where Cellgevity® significantly inhibited rat CYP3A4, CYP2B1/2B2 and CYP2C9 after a 7-day treatment period. Horn, Reichert (12) showed that CYP activity can be modulated by both dose and treatment duration. Pichard-Garcia, Hyland (20) reported that higher concentrations of eletriptan induced CYP3A in culture medium, however, lower concentrations did not cause CYP3A induction. Organisms after exposure to xenobiotics or foreign chemicals often develop adaptive mechanisms where they increase metabolism in an attempt to get rid of the insulting agent. This adaptive mechanism may have accounted for the increased activity of rat CYP1A1/2, CYP2C9 and CYP2D6 observed when relatively higher doses of Cellgevity® (38.63 mg/kg, 77.25 mg/kg and 154.50 mg/kg) were used in the current study as compared to our earlier study (4 mg/kg and 8 mg/kg of Cellgevity®). Indeed, it may not be entirely prudent to extrapolate animal studies to humans, but these data give credence to the fact that dietary supplements could modulate CYP enzymes in humans. If this increase in rat CYP1A1/2, CYP2C9 and CYP2D6 (dose-dependent in the case of CYP2C9) activity are of clinical relevance, then emphasis should be made on maximum therapeutic daily doses of Cellgevity® in humans. There are reports of potential interaction between dietary supplements/herbal products and conventional drugs. The commonest of these interactions appear to occur at the level of drug metabolism; especially with liver microsomal enzymes. We earlier reported that Cellgevity® significantly inhibited rat liver CYP3A4, CYP2C9 and CYP2B1/2B2 after a 7-day treatment period (11). On the premise that CYP3A4 was inhibited by Cellgevity® in our earlier research, the current study also sought to determine the effect of Cellgevity® on the pharmacokinetics of carbamazepine (a CYP3A4 substrate) after a 14-day treatment period in SD rats. The peak concentration for rats administered carbamazepine with Cellgevity® was about 2-fold greater than rats administered carbamazepine with saline. Total drug exposure at the last sample time point (AUC 0→24 ) was also about 2-fold greater in rats administered carbamazepine with Cellgevity® compared to carbamazepine with saline. This meant that there was a relatively slower elimination of carbamazepine in rats administered carbamazepine with Cellgevity®, possibly via inhibition of rat CYP 3A activity by Cellgevity®. This ultimately led to a longer half-life (2.3 h) among rats administered carbamazepine with Cellgevity®. Although the current study using an animal model showed some level of interaction between Cellgevity® and carbamazepine, a limitation was the inadequate sample volumes at each time point, which led to pooling of serum. Therefore, mean pharmacokinetic parameters within each group could not be obtained for statistical comparison. Notwithstanding, comparison of pharmacokinetic parameters of traditional versus pooled samples have found no statistical significant difference between the two sets of parameter estimates (19). It can, therefore, be inferred from the current study that Cellgevity® had some level of interaction with carbamazepine, possibly through inhibition of CYP3A, the enzyme known to metabolize carbamazepine. In a related study, anecdotal reports suggested that epileptic patients were taking diosmin, a widely used flavonoid in the treatment of varicose veins and haemorrhoids, along with carbamazepine. This led to a study to ascertain possible interaction between these two agents in an animal model (21). C max , AUC, and t 1/2 of carbamazepine were significantly elevated in diomin-treated rats compared to control rats (21). This, therefore, corroborates findings from the current study that there is the potential for herbal medicines, dietary supplements, and food to interact with conventional drugs in vivo (22), and that studies of this nature ought to be conducted to identify potential herb-drug interactions. Conclusion In the current study, Cellgevity® caused appreciable increase in the activities of rat liver CYP1A1/2, CYP2C9 and CYP2D6 enzymes after a 30-day treatment period. Additionally, Cellgevity® altered the pharmacokinetics (elimination rate and half-life) of carbamazepine in Sprague-Dawley rats after a 14-day treatment. Although this study was conducted in an animal model, this finding is noteworthy, as this may serve as a basis for future studies, i.e., assessing the effect of Cellgevity® on protein content and/or mRNA of distinct CYP proteins in rat livers and direct effect of Cellgevity® on recombinant human CYP enzymes. Declarations Ethics approval and consent to participate This research was approved by the College of Health Sciences Ethical and Protocol Review Committee (Protocol ID: CHS-Et/M.9 – P1.16/2017-2018) of the University of Ghana. Consent for publication Not applicable. Availability of data and materials Data used to support the findings of this study are available from the corresponding author upon request. Competing interests The authors declare that they have no competing interests. Funding This project was made possible with financial support from Building a New Generation of Academics in Africa (BANGA-Africa) Project (Grant Number: UG-BA/PD-006/2017-2018), University of Ghana, which is funded by the Carnegie Corporation of New York. The funding body, BANGA-Africa Project, played no role in the design of the study, data collection, data analysis, writing of the manuscript and in the payment of the article processing charge (APC). Authors’ contributions BBN and SKA conceived the study and designed the methodology. MA, SYA, AA, EAD performed experiments under the supervision of BBN, SKA, KFMO and RAO. SKA, BBN, KFMO, IJAG, RAO, MA, SYA, AA and EAD conducted the analysis of data. SKA, BBN, KFMO, IJAG and RAO drafted the manuscript. All authors read and approved the final version of the manuscript. Acknowledgement We would like to acknowledge Jessica Twumasi-Ankrah and all laboratory technicians at the School of Pharmacy and Noguchi Memorial Institute for Medical Research, University of Ghana, for their technical support. Abbreviations ANOVA: One-Way Analysis of Variance AUC: Area Under the Concentration-time Curve C max : Maximum Serum Drug Concentration CYP: Cytochrome P450 Enzymes DMSO: Dimethyl Sulfoxide FPIA: Fluorescence Polarization Immunoassay H-D: High Dose HPLC: High-Performance Liquid Chromatography K e : Elimination Rate Constant L-D: Low Dose M-D: Medium Dose NADPH: Nicotinamide Adenine Dinucleotide Phosphate N-C: Negative Control NCD: Non-Communicable Disease P-C: Positive Control rpm: Revolutions Per Minute SD: Sprague-Dawley t 1/2 : Elimination Half-life T max : Time to Reach Maximum Serum Drug Concentration References Valavanidis A. Dietary Supplements: Beneficial to Human Health or Just Peace of Mind? A Critical Review on the Issue of Benefit/Risk of Dietary Supplements. 2016. Pitetti R, Singh S, Hornyak D, Garcia S, Herr S. Complementary and alternative medicine use in children. Pediatric emergency care. 2001;17(3):165. Cott JM. Herb‐Drug Interactions: Theory versus Practice. Molecular nutrition food research. 2008;52(7):745-6. Ameade EPK, Ibrahim M, Ibrahim H-S, Habib RH, Gbedema SY. Concurrent Use of Herbal and Orthodox Medicines among Residents of Tamale, Northern Ghana, Who Patronize Hospitals and Herbal Clinics. Evidence-Based Complementary Aternative Medicine. 2018;2018. Oz HS, Chen TS, Nagasawa H. Comparative efficacies of 2 cysteine prodrugs and a glutathione delivery agent in a colitis model. Transl Res. 2007;150(2):122-9. Roberts JC, Charyulu RL, Zera RT, Nagasawa HT. Protection against acetaminophen hepatotoxicity by ribose-cysteine (RibCys). Pharmacol Toxicol. 1992;70(4):281-5. Chan WK, Delucchi AB. Resveratrol, a red wine constituent, is a mechanism-based inactivator of cytochrome P450 3A4. Life Sci. 2000;67(25):3103-12. Singh RP, Dhanalakshmi S, Rao AR. Chemomodulatory action of Aloe vera on the profiles of enzymes associated with carcinogen metabolism and antioxidant status regulation in mice. Phytomedicine. 2000;7(3):209-19. Nagasawa HT. Method to enhance delivery of glutathione and ATP levels in cells. Google Patents; 2015. Nebel A, Schneider BJ, Baker RK, Kroll DJ. Potential metabolic interaction between St. John's wort and theophylline. The Annals of pharmacotherapy. 1999;33(4):502. N’guessan BB, Amponsah SK, Dugbartey GJ, Awuah KD, Dotse E, Aning A, et al. In Vitro Antioxidant Potential and Effect of a Glutathione-Enhancer Dietary Supplement on Selected Rat Liver Cytochrome P450 Enzyme Activity. Evidence-Based Complementary and Alternative Medicine. 2018;2018. Horn TL, Reichert MA, Bliss RL, Malejka-Giganti D. Modulations of P450 mRNA in liver and mammary gland and P450 activities and metabolism of estrogen in liver by treatment of rats with indole-3-carbinol. Biochemical pharmacology. 2002;64(3):393-404. Spina E, Pisani F, Perucca E. Clinically significant pharmacokinetic drug interactions with carbamazepine. Clinical pharmacokinetics. 1996;31(3):198-214. Nair AB, Jacob S. A simple practice guide for dose conversion between animals and human. Journal of basic and clinical pharmacy. 2016;7(2):27. Appiah-Opong R, de Esch I, Commandeur JN, Andarini M, Vermeulen NP. Structure–activity relationships for the inhibition of recombinant human cytochromes P450 by curcumin analogues. European journal of medicinal chemistry. 2008;43(8):1621-31. Appiah-Opong R, Commandeur JN, van Vugt-Lussenburg B, Vermeulen NP. Inhibition of human recombinant cytochrome P450s by curcumin and curcumin decomposition products. Toxicology. 2007;235(1-2):83-91. Umegaki K, Saito K, Kubota Y, Sanada H, Yamada K, Shinozuka K. Ginkgo biloba extract markedly induces pentoxyresorufin O-dealkylase activity in rats. The Japanese Journal of Pharmacology. 2002;90(4):345-51. Busby WF, Jr., Ackermann JM, Crespi CL. Effect of methanol, ethanol, dimethyl sulfoxide, and acetonitrile on in vitro activities of cDNA-expressed human cytochromes P-450. Drug metabolism and disposition: the biological fate of chemicals. 1999;27(2):246-9. Riad LE, Chan KK, Sawchuk RJ. Determination of the relative formation and elimination clearance of two major carbamazepine metabolites in humans: a comparison between traditional and pooled sample analysis. Pharmaceutical research. 1991;8(4):541-3. Pichard-Garcia L, Hyland R, Baulieu J, Fabre J-M, Milton A, Maurel P. Human hepatocytes in primary culture predict lack of cytochrome P-450 3A4 induction by eletriptan in vivo. Drug metabolism and disposition. 2000;28(1):51-7. Bedada SK, Neerati P. Modulation of CYP3A enzyme activity by diosmin and its consequence on carbamazepine pharmacokinetics in rats. Naunyn-Schmiedeberg's archives of pharmacology. 2018;391(2):115-21. Fong SYK, Gao Q, Zuo Z. Interaction of carbamazepine with herbs, dietary supplements, and food: a systematic review. Evidence-Based Complementary Alternative Medicine. 2013;2013. Tables Table 1: Summary of the effect of Cellgevity® on selected rat CYP enzyme activity CYP ISOFORM ASSAY EFFECT OF CELLGEVITY® ON CYP ACTIVITY CYP3A4 BROD No significant increase in enzyme activity CYP2B1/2 PROD No significant increase in enzyme activity CYP1A1/2 MROD Significant increase in enzyme activity (H-D: p < 0.001) CYP2C9 Diclofenac Hydroxylation Significant increase in enzyme activity (L-D: p < 0.05; M-D and H-D: p < 0.001) CYP2D6 Dextromethorphan O-Demethylation Significant increase in enzyme activity (L-D and M-D: p < 0.05; H-D: p < 0.01) L-D = Low dose (38.63 mg/kg) of Cellgevity ® ; M-D = Medium dose (77.25 mg/kg) of Cellgevity ® ; H-D = High dose (154.50 mg/kg) of Cellgevity ® Table 2: Pharmacokinetic parameters of carbamazepine for the two groups of rats administered carbamazepine plus Cellgevity®, or carbamazepine plus normal saline over a period of 14 days. PARAMETERS (UNIT) CARBAMAZEPINE + NORMAL SALINE CARBAMAZEPINE + CELLGEVITY® C max (µmol/L) 11 20 T max (h) 4 4 k e (h -1 ) 0.41 0.28 t 1/2 (h) 1.7 2.3 AUC 0→24 (µmol.h/L) 170 347 AUC 0→ ∞ (µmol.h/L) 178 375 Cite Share Download PDF Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-10386","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research article","associatedPublications":[],"authors":[{"id":492989,"identity":"8ca5d75a-7c70-4534-b8c0-2dc08c599f19","order_by":1,"name":"Seth Kwabena Amponsah","email":"","orcid":"","institution":"University of Ghana School of Pharmacy","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Seth","middleName":"Kwabena","lastName":"Amponsah","suffix":""},{"id":492990,"identity":"2e51fd23-32c6-43f5-bdda-0b6502f27112","order_by":2,"name":"Benoit Banga Nguessan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA50lEQVRIiWNgGAWjYBACAwk4k/kAiGRsYGBgI1YLWwLJWngMiNNiLt17+DVPRZ09/+yeb5JfGGxkNxxgv/YAnxbLOefSrHnOHE6ccefsNmkZhjTjDQd4yg3wOuxGjpkxb9uBBIYbudukJRgOJwK1pEkQ1vKvzl7+Rs4zoJb/RGkxfszbwMy44UYOm+QHhgNALezH8GqxnHPGjHHOscOJG2+kGVszGCQbzzzMw4ZXi7l0j/GHNzV19nI3kh/e/FFhJ9t3vP0ZXi1AgDCTGRw1EBIvYP4AYzH+AFPsDwhpGQWjYBSMgpEFAJD3TMlztPv3AAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-0749-2218","institution":"University of Ghana School of Pharmacy","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Benoit","middleName":"Banga","lastName":"Nguessan","suffix":""},{"id":492991,"identity":"08d15610-4be9-4490-be5f-ffea50d3ee3c","order_by":3,"name":"Martin Akandawen","email":"","orcid":"","institution":"University of Ghana School of Pharmacy","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Martin","middleName":"","lastName":"Akandawen","suffix":""},{"id":492992,"identity":"5a21e772-b6e3-4492-9c49-9b0be5eb1a96","order_by":4,"name":"Abigail Aning","email":"","orcid":"","institution":"University of Ghana Noguchi Memorial Institute for Medical Research","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Abigail","middleName":"","lastName":"Aning","suffix":""},{"id":492993,"identity":"b04ae9e3-bb2c-4e81-9fb8-1fa62b44cda7","order_by":5,"name":"Sedem Yawa Agboli","email":"","orcid":"","institution":"University of Ghana School of Pharmacy","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Sedem","middleName":"Yawa","lastName":"Agboli","suffix":""},{"id":492994,"identity":"b12f5644-fa21-4b9c-b3ab-cccd5095b87a","order_by":6,"name":"Eunice Ampem Danso","email":"","orcid":"","institution":"University of Ghana Noguchi Memorial Institute for Medical Research","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Eunice","middleName":"Ampem","lastName":"Danso","suffix":""},{"id":492995,"identity":"a20982bb-17f4-4197-866a-6cfd01deaec6","order_by":7,"name":"Kwabena Frimpong-Manso Opuni","email":"","orcid":"","institution":"University of Ghana School of Pharmacy","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Kwabena","middleName":"Frimpong-Manso","lastName":"Opuni","suffix":""},{"id":492996,"identity":"0d6412b0-bfa6-4ce9-99b7-15ab253d2e3a","order_by":8,"name":"Isaac Julius Asiedu-Gyekye","email":"","orcid":"","institution":"University of Ghana School of Medicine and Dentistry","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Isaac","middleName":"Julius","lastName":"Asiedu-Gyekye","suffix":""},{"id":492997,"identity":"021e3bce-7a67-4d27-8ed6-829863e4641d","order_by":9,"name":"Regina Appiah-Opong","email":"","orcid":"","institution":"University of Ghana Noguchi Memorial Institute for Medical Research","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Regina","middleName":"","lastName":"Appiah-Opong","suffix":""}],"badges":[],"createdAt":"2019-12-26 12:54:32","currentVersionCode":2,"declarations":"","doi":"10.21203/rs.2.19774/v2","doiUrl":"https://doi.org/10.21203/rs.2.19774/v2","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":928913,"identity":"3a92cd84-bb18-465e-b47d-35b27c3b8feb","added_by":"auto","created_at":"2020-04-20 15:02:50","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":71533,"visible":true,"origin":"","legend":"Rat liver CYP2C9 activity for various treatment groups after 30-day administration: N-C = Negative control; P-C = Positive control (Phenobarbital); L-D = Low dose (38.63 mg/kg) of Cellgevity®; M-D = Medium dose (77.25 mg/kg) of Cellgevity®; H-D = High dose (154.50 mg/kg) of Cellgevity®. Data represents mean ± standard deviations. ∗ and ∗∗∗ represent values significantly different from the negative control as indicated by 𝑝 \u003c 0.05 and 𝑝 \u003c 0.001, respectively","description":"","filename":"fig1.PNG","url":"https://assets-eu.researchsquare.com/files/rs-10386/v2/fig1.PNG"},{"id":928914,"identity":"0638780a-c05b-4968-b018-ae17bcb38d25","added_by":"auto","created_at":"2020-04-20 15:02:50","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":61159,"visible":true,"origin":"","legend":"Rat liver CYP2D6 activity for various treatment groups after 30 day administration: N-C = Negative control; P-C = Positive control (Phenobarbital); L-D = Low dose (38.63 mg/kg) of Cellgevity®; M-D = Medium dose (77.25 mg/kg) of Cellgevity®; H-D = High dose (154.50 mg/kg) of Cellgevity®. Data represents mean ± standard deviations. ∗ and ∗∗ represent values significantly different from the negative control as indicated by 𝑝 \u003c 0.05 and 𝑝 \u003c 0.01, respectively","description":"","filename":"fig2.PNG","url":"https://assets-eu.researchsquare.com/files/rs-10386/v2/fig2.PNG"},{"id":928915,"identity":"06f01b61-a854-49df-a568-7009071c8a4b","added_by":"auto","created_at":"2020-04-20 15:02:50","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":66228,"visible":true,"origin":"","legend":"Rat liver CYP1A1/2 activity for various treatment groups after 30 day administration: N-C = Negative control; P-C = Positive control (Phenobarbital); L-D = Low dose (38.63 mg/kg) of Cellgevity®; M-D = Medium dose (77.25 mg/kg) of Cellgevity®; H-D = High dose (154.50 mg/kg) of Cellgevity®. Data represent mean ± standard deviations. ∗∗∗ represents values significantly different from the negative control as indicated by 𝑝 \u003c 0.001, and ns means not significantly different","description":"","filename":"fig3.PNG","url":"https://assets-eu.researchsquare.com/files/rs-10386/v2/fig3.PNG"},{"id":928916,"identity":"851fe421-84a2-4394-8f0e-142a9336db32","added_by":"auto","created_at":"2020-04-20 15:02:50","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":68359,"visible":true,"origin":"","legend":"Rat liver CYP2B1/2 activity for various treatment groups after 30 day administration: N-C = Negative control; P-C = Positive control (Phenobarbital); L-D = Low dose (38.63 mg/kg) of Cellgevity®; M-D = Medium dose (77.25 mg/kg) of Cellgevity®; H-D = High dose (154.50 mg/kg) of Cellgevity®. Data represent mean ± standard deviations. ∗∗∗ represents values significantly different from the negative control group as indicated by 𝑝 \u003c 0.001","description":"","filename":"fig4.PNG","url":"https://assets-eu.researchsquare.com/files/rs-10386/v2/fig4.PNG"},{"id":928917,"identity":"e40f0fca-0ce4-4f60-9cd1-650bca9a2760","added_by":"auto","created_at":"2020-04-20 15:02:50","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":69322,"visible":true,"origin":"","legend":"Rat liver CYP3A4 activity for various treatment groups after 30 day administration: N-C = Negative control; P-C = Positive control (Phenobarbital); L-D = Low dose (38.63 mg/kg) of Cellgevity®; M-D = Medium dose (77.25 mg/kg) of Cellgevity®; H-D = High dose (154.50 mg/kg) of Cellgevity®. Data represents mean ± standard deviations. ∗∗∗ represents values significantly different as indicated by 𝑝 \u003c 0.001","description":"","filename":"fig5.PNG","url":"https://assets-eu.researchsquare.com/files/rs-10386/v2/fig5.PNG"},{"id":928918,"identity":"b0275335-31f4-4a9e-81c3-f3c72204d327","added_by":"auto","created_at":"2020-04-20 15:02:50","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":35577,"visible":true,"origin":"","legend":"Concentration-time curves of carbamazepine with Cellgevity® (grey solid line; filled squares) and carbamazepine with normal saline (red solid line; filled circles). Serum samples from rats in the same group (n = 6) were pooled together at each sampling time point.","description":"","filename":"fig6.PNG","url":"https://assets-eu.researchsquare.com/files/rs-10386/v2/fig6.PNG"},{"id":13499107,"identity":"4fea6ba5-90d5-42d1-a9c4-860d54134817","added_by":"auto","created_at":"2021-09-16 23:00:45","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1525359,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-10386/v2/4c4f239e-a27e-40b3-a5d8-ebf2d95d0895.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eEffect of Cellgevity® Supplement on Selected Rat Liver Cytochrome P450 Enzyme Activity and Pharmacokinetic Parameters of Carbamazepine\u003c/p\u003e","fulltext":[{"header":"Background","content":"\u003cp\u003eDietary supplements may be vitamins, minerals or herbal products that are known to improve the well-being of humans (1). This clearly denotes the use of these supplements as an addition to dietary requirements that may not be met by daily meals. However, under no circumstance should dietary supplements be used as replacement for daily meals. The United States Food and Drugs Administration also prohibits the indication of dietary supplements as treatment for diseases or any indication that connotes dietary supplements as primary therapeutic agents (1).\u003c/p\u003e\n\u003cp\u003eWith increase in the incidence of non-communicable diseases (NCDs) such as diabetes, cardiomyopathies, cancers and epilepsy, which are often associated with oxidative stress, people resort to the use of dietary supplements (known antioxidants) to prevent these diseases (2). Interestingly, some individuals who use dietary supplements have the notion that these agents may enhance the effects of conventional drugs (3). This assertion may be false. It is noteworthy, however, that synergy between some dietary supplements and conventional drugs have been reported (4).\u003c/p\u003e\n\u003cp\u003eCurrently available on the market are a number of dietary supplements known to replenish levels of reduced glutathione, a free radical scavenger, in cells of humans. One such supplement is Cellgevity\u0026reg; which contains a glutathione precursor molecule, riboceine (D-ribose-L-cysteine). Riboceine is known to deliver cysteine into cells and enhance reduced glutathione level in the body (5, 6). The other constituents of Cellgevity\u0026reg; are broccoli seed extract, turmeric root extract, resveratrol, grape seed extract, quercetin, curcumin, milk thistle, vitamin C, selenomethionine, cordyceps, black pepper and aloe extract. Some of these constituents of Cellgevity\u0026reg; are known inducers and/or inhibitors of cytochrome P450 (CYP) enzymes (7, 8). Cellgevity\u0026reg; is marketed and distributed by Max International, which has branches in 14\u0026nbsp;countries\u0026nbsp;(United States, Nigeria, Cote d\u0026rsquo;Ivoire, New Zealand, Singapore, Costa Rica, Columbia, Philippines, El Salvador, Malaysia, Guatemala, Ghana and Hong Kong). Cellgevity\u0026reg; has gained popularity in these countries due to the fact that patronizers know that the supplement has a high antioxidant potential (9).\u003c/p\u003e\n\u003cp\u003eReports suggest that there could be clinically important modulation of CYP enzyme activity by supplements and/or herbal products. This could result in adverse or sub-therapeutic effects of concurrently administered conventional drugs. For example, St. John\u0026rsquo;s wort was reported to decrease the serum concentration of theophylline (a bronchodilator) as a result of CYP enzyme induction (10). This interaction between St John\u0026rsquo;s wort and theophylline could lead to sub-therapeutic effect of normal doses of theophylline when there is co-administration. For this reason, patients are often advised not to take theophylline concomitantly with St. John\u0026rsquo;s wort. \u003cstrong\u003eMost of the individual constituents of Cellgevity\u0026reg; can modulate xenobiotic metabolizing enzyme activity, and several studies have reported these. However, there is paucity of scientific data on the effect of all these constituents (extracts) combined in Cellgevity\u0026reg; on xenobiotic metabolizing enzymes (CYP enzymes)\u003c/strong\u003e\u003cstrong\u003e.\u003c/strong\u003e In our earlier study, we reported that Cellgevity\u0026reg; at 4 and 8 mg/kg significantly inhibited rat liver CYP2C9, CYP2B1/2B2 and CYP3A4 after a 7-day treatment period (11). In animal models, it is relevant that doses of test agents (drugs/food supplements/herbal products) are at equivalent doses of what pertains in humans. Therefore, as a follow-up to our earlier study, we decided to use animal equivalent doses of Cellgevity\u0026reg; after scaling doses used in humans (12) in the current study. We also decided to administer Cellgevity\u0026reg; \u0026nbsp;over a longer period of time, i.e. 30 days.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eWe have previously reported that Cellgevity\u003csup\u003e\u0026reg;\u003c/sup\u003e inhibited rat liver CYP3A4 after a 7-day treatment period \u003c/strong\u003e\u003cstrong\u003e(11)\u003c/strong\u003e\u003cstrong\u003e. We, therefore, sought to elucidate possible effect of Cellgevity\u003csup\u003e\u0026reg;\u003c/sup\u003e on carbamazepine, also known to be extensively metabolized by CYP3A4. This was to aid in assessing potential interaction between these two agents.\u003c/strong\u003e Carbamazepine is one of the most commonly prescribed drugs in the management of epilepsy. Due to the chronic nature of epilepsy, and the fact that patients have to take carbamazepine for a long time (lifetime in most cases), there is the potential for clinically significant interactions between carbamazepine and co-administered agents like dietary supplements, herbal products and food (13). Therefore, in a parallel experiment, the effect of Cellgevity\u0026reg; on the pharmacokinetics of carbamazepine (a CYP3A4 substrate), was investigated.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003e\u003cstrong\u003eAnimal care and safety\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was approved by the College of Health Sciences Ethical and Protocol Review Committee (Protocol ID: CHS-Et/M.9 \u0026ndash; P1.16/2017-2018) of the University of Ghana. All animal procedures used in this study were in accordance with the National Institute of Health Guidelines for the Care and Use of Laboratory Animals.\u003c/p\u003e\n\u003cp\u003eMale Sprague-Dawley (SD) rats, weighing 150-200 g and 6-8 weeks old, were obtained from the Center for Plant Medicine Research, Mampong, Eastern Region, Ghana. The animals were housed in stainless steel cages. Each rat occupied a minimum space of 2 cubic feet (61 cm x 31 cm x 31 cm) with softwood shavings as bedding for their comfort. The SD rats were fed with normal pellet diet (AGRIMAT, Kumasi, Ghana), given water \u003cem\u003ead libitum\u003c/em\u003e, and maintained under standard laboratory conditions (temperature ~25\u0026deg;C, relative humidity 60-70%, and 12 h light-dark cycle). The animals\u0026rsquo; feeding area and water troughs were cleaned regularly to prevent contamination. Animals were acclimatized under stated conditions for 7 days before experiment commenced.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHepatic enzyme induction/inhibition studies\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eAnimal grouping and treatment administration\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eIn determining the effect of Cellgevity\u0026reg; on CYP enzymes after a 30-day treatment period, male SD rats were put into five groups (6 rats in each group). All treatments were \u003cem\u003eper os\u003c/em\u003e and for 30 days. Group 1 was administered distilled water, the vehicle used in dissolving Cellgevity\u0026reg; (purchased from Max International, Ghana), and that served as the negative control (N-C) group. Groups 2, 3 and 4 received daily a low dose (L-D) of 38.63 mg/kg Cellgevity\u0026reg;, medium dose (M-D) of 77.25 mg/kg Cellgevity\u0026reg; and high dose (H-D) of 154.50 mg/kg Cellgevity\u0026reg;, respectively. The doses of Cellgevity\u0026reg; administered to SD rats were animal equivalents of what pertains in humans, and calculated as described by Nair AB and Jacob SA (14). The human dose of Cellgevity\u0026reg; is 12.46 mg/kg \u003cem\u003eper os\u003c/em\u003e. The SD rats in Group 5 received an oral dose of phenobarbital (Kinapharma, Ghana) 15 mg/kg daily, and that served as the positive control (P-C). After the 30-day treatment period, animals were sacrificed by cervical dislocation. Livers were excised, washed in ice-cold saline solution, weighed, and stored at -80\u003csup\u003eo\u003c/sup\u003eC until use.\u003c/p\u003e\n\u003ch2\u003e\u003cem\u003e\u003cu\u003eMicrosomal Preparation\u003c/u\u003e\u003c/em\u003e\u003c/h2\u003e\n\u003cp\u003eLivers were thawed and homogenized in potassium phosphate buffer (pH 7.4) using a mortar and pestle on ice. Homogenized samples were first centrifuged at 4,000 rpm for 20 min. The supernatant obtained was re-centrifuged (Beckman Avanti J-25, USA) at 25,000 rpm for 2 h. The pellets obtained, microsomes, were collected and stored at -80\u003csup\u003eo\u003c/sup\u003eC until use.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eCYP2C9 (\u003c/u\u003e\u003c/em\u003e\u003cem\u003e\u003cu\u003eDiclofenac Hydroxylation) \u003c/u\u003e\u003c/em\u003e\u003cem\u003e\u003cu\u003eand CYP2D6 (\u003c/u\u003e\u003c/em\u003e\u003cem\u003e\u003cu\u003eDextromethorphan O-demethylation)\u003c/u\u003e\u003c/em\u003e\u003cem\u003e\u003cu\u003e Assays\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe assays were performed as previously described (15), with some modification. A volume of 350 \u0026micro;L of 0.1M potassium phosphate buffer (pH 7.4), 50 \u0026micro;L of 1 mM substrate (diclofenac for CYP2C9 assay and dextromethorphan for CYP2D6 \u0026ndash; both substrates purchased from Sigma-Aldrich, USA) and 50 \u0026micro;L of 2.5 mg/mL microsome (obtained from rat livers from respective groups) were mixed separately in Eppendorf tubes. The mixtures were pre-incubated at 37\u003csup\u003eo\u003c/sup\u003eC for 5 min. A volume of 50 \u0026micro;L of 1 mM nicotinamide adenine dinucleotide phosphate (NADPH) [Sigma-Aldrich, USA] was added, mixed and incubated at 37\u003csup\u003eo\u003c/sup\u003eC for 45 min. A 100 \u0026micro;L stopping solution (ZnSO\u003csub\u003e4\u003c/sub\u003e.7H\u003csub\u003e2\u003c/sub\u003eO) was added and the mixture centrifuged at 4000 rpm for 5 min. The supernatants were aliquoted into High-Performance Liquid Chromatography (HPLC, Shimadzu, Japan) vials.\u003c/p\u003e\n\u003cp\u003eSamples were analyzed using HPLC. The chromatographic system consisted of a binary solvent delivery system (LC - 20AB), a degasser (DGU-20A3), an auto-sampler (SIL - 20ACHT), a column temperature controller (CTO - 10AS VP) and a photodiode array detector (SPD - M20A) for CYP2C9 metabolites and fluorescence detector (RF - 10A \u003csub\u003eXL\u003c/sub\u003e) for CYP2D6 metabolites. The following chromatographic conditions were used for the analysis of CYP2C9; column, C18 (Shimadzu, Japan), diameter 5 \u0026mu;m, length x width 150 mm \u0026times; 4.6 mm; flow rate, 1 mL/min; column temperature, 40\u003csup\u003eo\u003c/sup\u003eC; injection volume, 20 \u0026mu;L; mobile phase, 20 mM potassium phosphate buffer (pH 7.4)/methanol/acetonitrile (60:22.5:17.5, v/v/v). The same chromatographic conditions were used for the analysis of the CYP2D6, with modification to the mobile phase where \u003cstrong\u003e3 solvents\u003c/strong\u003e were used (acetonitrile/distilled water/triethylamine; 24:75:1, v/v/v).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eCYP1A1/1A2 - Ethoxyresorufin O-deethylase (EROD), CYP2B1/2B2 -\u003c/u\u003e\u003c/em\u003e\u003cu\u003e\u003cem\u003ePentoxyresorufin O-depentylase (PROD) and CYP3A4 - Benzyloxyresorufin O-debenzylase (BROD) Assays\u003c/em\u003e\u003c/u\u003e\u003c/p\u003e\n\u003cp\u003eThe assays were performed as previously described (16, 17), with some modification. In brief, microsomes (CYP enzymes) were tested in a total volume of 100 \u0026mu;L. Aliquots of 70 \u0026mu;L potassium phosphate buffer (pH 7.4) were placed into 96-well black plates. This was followed by addition of 10 \u0026mu;L of 50 \u0026mu;M substrate concentration (resorufin ethyl ether for CYP1A1/2, pentoresorufin for CYP2B1/2 and resorufin benzyl ether for CYP3A4; all substrates purchased from Sigma-Aldrich, USA). The final substrate concentration in 100 \u0026mu;L total reaction volume was 5 \u0026mu;M with 0.25% (v/v) dimethyl sulfoxide (DMSO). It is noteworthy that CYP activities were not expected to be affected at the DMSO concentration used in this experiment (18). Aliquots of 10 \u0026mu;L enzyme (microsome from each rat liver from respective Groups) corresponding to 1 mg/mL protein concentration and the vehicle were added in triplicates. The mixtures were pre-incubated at 37\u003csup\u003eo\u003c/sup\u003eC for 5 min. A volume of 10 \u0026mu;L of NADPH was then added to each well and the setup was incubated for 10 min for CYP1A1/2, 20 min for CYP2B1/2 and 30 min for CYP3A4 assays, respectively. Aliquots of 40 \u0026mu;L of stopping solution (20% 0.5 M Tris: 80% acetonitrile) were added to each well and shaken gently. Fluorescence of wells was read at wavelengths of 530 nm excitation and 586 nm emission. Triplicate experiments were performed. The average absorbance of the blank was subtracted from the average absorbance of each sample.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFourteen-day treatment of \u003c/strong\u003e\u003cstrong\u003eCellgevity\u0026reg; on the pharmacokinetics of carbamazepine in rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eAnimal grouping and treatment administration\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eTwelve male SD rats were obtained for this aspect of the study. The animals were put into 2 groups of 6 (Group 1 and Group 2). All treatments were \u003cstrong\u003e\u003cem\u003eper os\u003c/em\u003e\u003c/strong\u003e and for 14 days. Group 1 was administered carbamazepine plus saline and Group 2, Cellgevity\u0026reg; plus carbamazepine. A dose of 77.25 mg/kg/day of Cellgevity\u0026reg; plus 80 mg/kg of carbamazepine, both equivalent doses per serving/administration in humans scaled to SD rats (14), were administered to rats in Group 2. Rats in Group 1 received 80 mg/kg/day of carbamazepine plus normal saline (same volume as calculated per rat for the Cellgevity\u0026reg; dose).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eBlood sample collection\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eAfter administration of carbamazepine with or without Cellgevity\u0026reg; every 24 h for 14 consecutive days to Groups 1 and 2, tail vein blood samples were taken following the dose administered on the 14\u003csup\u003eth\u003c/sup\u003e day. Samples were drawn after 0.5, 1, 4, 12 and 24 h. Blood was collected into microtainer gel tubes and centrifuged at 2000 rpm for 5 min to separate serum, and this was stored at -20\u0026ordm;C until analysis.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eAssay for carbamazepine in serum\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDue to low sample volumes from tail veins of SD rats, serum samples from rats in the same group (6 animals) at each time point were pooled together. Such that, for instance, serum samples of Group 1 SD rats at time 0.5, 1, 4, 12 or 24 h, were pooled together to obtain a single sample. Usually, challenges with low sample volume can be circumvented by the approach of sample-pooling \u003c/strong\u003e\u003cstrong\u003e(19)\u003c/strong\u003e\u003cstrong\u003e.\u003c/strong\u003e Analysis of carbamazepine in serum was by Fluorescence Polarization Immunoassay (FPIA) [Cobas Integra\u0026reg; 400 Plus, Roche, Philippines]. \u003cstrong\u003eThe lower limit of quantification of serum carbamazepine concentration was 0.8 \u0026micro;mol/L and coefficient of variation was \u0026lt; 5% over the entire calibration range.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eStatistical analysis\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eCYP activity of treatment groups was expressed as a percentage relative to the negative control group. All values were expressed as mean \u0026plusmn; standard deviation. Differences between groups were tested for significance using a One-Way Analysis of Variance (ANOVA). This was followed by post-hoc analysis using Bonferroni\u0026rsquo;s Multiple Comparison Tests. \u003cem\u003eP\u003c/em\u003e-values \u0026lt; 0.05 were considered statistically significant.\u003c/p\u003e\n\u003cp\u003eNon-compartmental pharmacokinetic analysis was used to estimate the various pharmacokinetic parameters of carbamazepine. The maximum serum drug concentration (C\u003csub\u003emax\u003c/sub\u003e) and its corresponding time (T\u003csub\u003emax\u003c/sub\u003e) were determined by visual inspection of the concentration-time curve. The linear trapezoidal rule was applied in extrapolating area under the concentration-time curves (AUCs) for the two groups. \u003cstrong\u003eThe elimination rate constant (K\u003csub\u003ee\u003c/sub\u003e) for both groups were extrapolations (apparent slope) from the last sample time point, i.e. 24 hours. K\u003csub\u003ee\u003c/sub\u003e for both groups was used to calculate corresponding elimination half-lives (t\u003csub\u003e1/2\u003c/sub\u003e)\u003c/strong\u003e\u003cstrong\u003e.\u003c/strong\u003e\u003cstrong\u003ePharmacokinetic analysis was conducted using GraphPad Prism 7.0.\u003c/strong\u003e\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eHepatic enzyme induction/inhibition studies\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll CYPs enzyme activities in the treatment groups were estimated relative to the negative control (N-C) group.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eCYP2C9 activity in SD rats after 30-day treatment\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eCYP2C9 enzyme activity was higher in the phenobarbital- and Cellgevity\u0026reg;-treated groups in comparison with the N-C group. The phenobarbital- and Cellgevity\u0026reg;-treated groups were significantly different from the N-C group. The increase in rat CYP2C9 enzyme activity by Cellgevity\u0026reg; was dose-dependent. Presentation of the effect of Cellgevity\u0026reg; on rat CYP2C9 enzyme activity in various groups is shown in Figure 1.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eCYP2D6 activity in SD rats after 30-day treatment\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eCYP2D6 enzyme activity was about 2.5-fold higher in the phenobarbital-treated and about 2-fold higher in the Cellgevity\u0026reg;-treated groups in comparison with the N-C group. The positive control (P-C) and H-D Cellgevity\u0026reg;-treated groups differed significantly (p \u0026lt; 0.01) from the N-C group. The L-D and M-D groups also differed significantly (p \u0026lt; 0.05) in comparison to the N-C group. \u003cstrong\u003eThe increase in rat CYP2D6 enzyme activity by Cellgevity\u0026reg; was not dose-dependent.\u003c/strong\u003e CYP2D6 enzyme activity of SD rats in various groups after 30-day treatment period is shown in Figure 2.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eCYP1A1/2 \u003c/u\u003e\u003c/em\u003e\u003cem\u003e\u003cu\u003eactivity in SD rats after 30-day treatment\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eCYP1A1/2 enzyme activity was higher in the phenobarbital- and Cellgevity\u0026reg;-treated groups in comparison with the N-C group. The Cellgevity\u0026reg;-treated L-D and M-D groups showed elevated CYP activity compared to N-C group, but these differences were not statistically significant. However, there was a significant difference between the H-D Cellgevity\u0026reg;-treated group and the N-C group. There was somewhat a dose-dependent increase in rat CYP1A1/2 enzyme activity in the Cellgevity\u0026reg;-treated group. CYP1A1/2 enzyme activity of SD rats in various groups after the 30-day treatment is shown in Figure 3.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eCYP2B1/2\u003c/u\u003e\u003c/em\u003e\u003cem\u003e\u003cu\u003eactivity in SD rats after 30-day treatment\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eCYP2B1/2 enzyme activity was elevated in the phenobarbital- and Cellgevity\u0026reg;-treated groups in comparison with the N-C group. The Cellgevity\u0026reg;-treated groups showed elevated levels compared to N-C group, but the differences were not statistically significant. There was also no dose-dependent effect of Cellgevity\u0026reg; on rat CYP2B1/2 enzyme activity. CYP2B1/2 enzyme activity of SD rats in various groups after 30-day treatment is shown in Figure 4.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eCYP3A4 \u003c/u\u003e\u003c/em\u003e\u003cem\u003e\u003cu\u003eactivity in SD rats after 30-day treatment\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eCYP3A4 enzyme activity was elevated in the phenobarbital- and Cellgevity\u0026reg;-treated groups in comparison with the N-C group. The Cellgevity\u0026reg;-treated groups showed elevated levels compared to the N-C group, but the differences were not statistically significant. There was also no dose-dependent effect of Cellgevity\u0026reg; on rat CYP3A4 enzyme activity. CYP3A4 enzyme activity of SD rats in various groups after 30-day treatment are shown in Figure 5.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cu\u003eOverall Effect of Cellgevity\u0026reg; on Rat CYP Enzyme Activity\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eAfter 30 days, when the Cellgevity\u0026reg;-treated groups were compared to the N-C group, the activities of CYP3A4 and CYP2B1/2 did not differ significantly. However, CYP1A1/2, CYP2C9 and CYP2D6 activities in SD rats treated with Cellgevity\u0026reg; were significantly increased compared to the N-C group. Additionally, the increase in CYP2C9 activity was dose-dependent. The overall effect of Cellgevity\u0026reg; on selected rat CYP enzymes is shown in Table 1.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFourteen-day treatment of Cellgevity\u0026reg; on the pharmacokinetics of carbamazepine in rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCarbamazepine concentration-time curves of rats administered carbamazepine with Cellgevity\u0026reg;, and carbamazepine with normal saline are shown in Figure 6. From the concentration-time curves, rats administered carbamazepine with Cellgevity\u0026reg; had a higher peak concentration at 4 h compared to the rats administered carbamazepine with normal saline.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eThe total carbamazepine exposure (AUC\u003csub\u003e0\u0026rarr;24\u003c/sub\u003e) and peak carbamazepine concentration for rats administered carbamazepine with normal saline were 170 \u0026micro;mol.h/L and 11 \u0026micro;mol/L, respectively, as against 347 \u0026micro;mol.h/L and 20 \u0026micro;mol/L, respectively, for rats administered carbamazepine with Cellgevity\u0026reg;. Elimination rate constants were 0.28 h\u003csup\u003e-1\u003c/sup\u003e for SD rats administered carbamazepine with Cellgevity\u0026reg; and 0.41 h\u003csup\u003e-1\u003c/sup\u003e for SD rats administered carbamazepine with normal saline. \u003c/strong\u003ePharmacokinetic parameters obtained from the concentration-time curves for the two groups are shown in Table 2.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eWe earlier reported the potential of Cellgevity\u0026reg; to modulate CYP enzymes in rats (11). In that study, 4 mg/kg and 8 mg/kg of Cellgevity\u0026reg; were admistered to SD rats over a period of 7 days. Cellgevity\u0026reg; inhibited rat liver CYP3A4, CYP2C9 and CYP1A2, after the 7-day treatment period (11). In the current study, an equivalent dose of Cellgevity\u0026reg; per serving in humans (12.46 mg/kg) was scaled to SD rats (14), and three doses used over a period of 30 days. Reports suggest that xenobiotics can modulate CYP enzymes depending on dose and treatment duration (12). This study, therefore, sought to investigate the effect of Cellgevity\u0026reg; on rat liver CYP enzymes using 38.63 mg/kg, 77.25 mg/kg and 154.50 mg/kg of Cellgevity\u0026reg; calculated after scaling from humans, and administering Cellgevity\u0026reg; for 30 days.\u003c/p\u003e\n\u003cp\u003eIn the present study (after the 30-day treatment period), Cellgevity\u0026reg; significantly increased the activity of rat CYP1A1/2, CYP2C9 and CYP2D6. These results differ from what we earlier reported (11), where Cellgevity\u0026reg; significantly inhibited rat CYP3A4, CYP2B1/2B2 and CYP2C9 after a 7-day treatment period. Horn, Reichert (12) showed that CYP activity can be modulated by both dose and treatment duration. Pichard-Garcia, Hyland (20) reported that higher concentrations of eletriptan induced CYP3A in culture medium, however, lower concentrations did not cause CYP3A induction. Organisms after exposure to xenobiotics or foreign chemicals often develop adaptive mechanisms where they increase metabolism in an attempt to get rid of the insulting agent. This adaptive mechanism may have accounted for the increased activity of rat CYP1A1/2, CYP2C9 and CYP2D6 observed when relatively higher doses of Cellgevity\u0026reg; (38.63 mg/kg, 77.25 mg/kg and 154.50 mg/kg) were used in the current study as compared to our earlier study (4 mg/kg and 8 mg/kg of Cellgevity\u0026reg;).\u003c/p\u003e\n\u003cp\u003eIndeed, it may not be entirely prudent to extrapolate animal studies to humans, but these data give credence to the fact that dietary supplements could modulate CYP enzymes in humans. If this increase in rat CYP1A1/2, CYP2C9 and CYP2D6 (dose-dependent in the case of CYP2C9) activity are of clinical relevance, then emphasis should be made on maximum therapeutic daily doses of Cellgevity\u0026reg; in humans.\u003c/p\u003e\n\u003cp\u003eThere are reports of potential interaction between dietary supplements/herbal products and conventional drugs. The commonest of these interactions appear to occur at the level of drug metabolism; especially with liver microsomal enzymes. We earlier reported that Cellgevity\u0026reg; significantly inhibited rat liver CYP3A4, CYP2C9 and CYP2B1/2B2 after a 7-day treatment period (11). On the premise that CYP3A4 was inhibited by Cellgevity\u0026reg; in our earlier research, the current study also sought to determine the effect of Cellgevity\u0026reg; on the pharmacokinetics of carbamazepine (a CYP3A4 substrate) after a 14-day treatment period in SD rats. \u003cstrong\u003eThe peak concentration for rats administered carbamazepine with Cellgevity\u0026reg; was about 2-fold greater than rats administered carbamazepine with saline. Total drug exposure at the last sample time point (AUC\u003csub\u003e0\u0026rarr;24\u003c/sub\u003e) was also about 2-fold greater in rats administered carbamazepine with Cellgevity\u0026reg; compared to carbamazepine with saline.\u003c/strong\u003e\u003cstrong\u003eThis meant that there was a relatively slower elimination of carbamazepine in rats administered carbamazepine with Cellgevity\u0026reg;, possibly via inhibition of rat CYP 3A activity by Cellgevity\u0026reg;.\u003c/strong\u003e This ultimately led to a longer half-life (2.3 h) among rats administered carbamazepine with Cellgevity\u0026reg;. \u003cstrong\u003eAlthough the current study using an animal model showed some level of interaction between Cellgevity\u0026reg; and carbamazepine, a limitation was the inadequate sample volumes at each time point, which led to pooling of serum. Therefore, mean pharmacokinetic parameters within each group could not be obtained for statistical comparison. Notwithstanding, comparison of pharmacokinetic parameters of traditional versus pooled samples have found no statistical significant difference between the two sets of parameter estimates (19).\u003c/strong\u003e It can, therefore, be inferred from the current study that Cellgevity\u0026reg; had some level of interaction with carbamazepine, possibly through inhibition of CYP3A, the enzyme known to metabolize carbamazepine.\u003c/p\u003e\n\u003cp\u003eIn a related study, anecdotal reports suggested that epileptic patients were taking diosmin, a widely used flavonoid in the treatment of varicose veins and haemorrhoids, along with carbamazepine. This led to a study to ascertain possible interaction between these two agents in an animal model (21). C\u003csub\u003emax\u003c/sub\u003e, AUC, and t\u003csub\u003e1/2\u003c/sub\u003e of carbamazepine were significantly elevated in diomin-treated rats compared to control rats (21). This, therefore, corroborates findings from the current study that there is the potential for herbal medicines, dietary supplements, and food to interact with conventional drugs \u003cem\u003ein vivo \u003c/em\u003e(22), and that studies of this nature ought to be conducted to identify potential herb-drug interactions.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn the current study, Cellgevity\u0026reg; caused appreciable increase in the activities of rat liver CYP1A1/2, CYP2C9 and CYP2D6 enzymes after a 30-day treatment period. Additionally, Cellgevity\u0026reg; altered the pharmacokinetics (elimination rate and half-life) of carbamazepine in Sprague-Dawley rats after a 14-day treatment. \u003cstrong\u003eAlthough this study was conducted in an animal model, this finding is noteworthy, as this may serve as a basis for future studies, i.e., assessing the effect of\u0026nbsp; Cellgevity\u0026reg; on protein content and/or mRNA of distinct CYP proteins in rat livers and direct effect of Cellgevity\u0026reg; on recombinant human CYP enzymes.\u003c/strong\u003e\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was approved by the College of Health Sciences Ethical and Protocol Review Committee (Protocol ID: CHS-Et/M.9 \u0026ndash; P1.16/2017-2018) of the University of Ghana.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData used to support the findings of this study are available from the corresponding author upon request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis project was made possible with financial support from Building a New Generation of Academics in Africa (BANGA-Africa) Project (Grant Number: UG-BA/PD-006/2017-2018), University of Ghana, which is funded by the Carnegie Corporation of New York. The funding body, BANGA-Africa Project, played no role in the design of the study, data collection, data analysis, writing of the manuscript and in the payment of the article processing charge (APC).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBBN and SKA conceived the study and designed the methodology. MA, SYA, AA, EAD performed experiments under the supervision of BBN, SKA, KFMO and RAO. SKA, BBN, KFMO, IJAG, RAO, MA, SYA, AA and EAD conducted the analysis of data. SKA, BBN, KFMO, IJAG and RAO drafted the manuscript. All authors read and approved the final version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe would like to acknowledge Jessica Twumasi-Ankrah and all laboratory technicians at the School of Pharmacy and Noguchi Memorial Institute for Medical Research, University of Ghana, for their technical support.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eANOVA: One-Way Analysis of Variance\u003c/p\u003e\n\u003cp\u003eAUC: Area Under the Concentration-time Curve\u003c/p\u003e\n\u003cp\u003eC\u003csub\u003emax\u003c/sub\u003e: Maximum Serum Drug Concentration\u003c/p\u003e\n\u003cp\u003eCYP: Cytochrome P450 Enzymes\u003c/p\u003e\n\u003cp\u003eDMSO: Dimethyl Sulfoxide\u003c/p\u003e\n\u003cp\u003eFPIA: Fluorescence Polarization Immunoassay\u003c/p\u003e\n\u003cp\u003eH-D: High Dose\u003c/p\u003e\n\u003cp\u003eHPLC: High-Performance Liquid Chromatography\u003c/p\u003e\n\u003cp\u003eK\u003csub\u003ee\u003c/sub\u003e: Elimination Rate Constant\u003c/p\u003e\n\u003cp\u003eL-D: Low Dose\u003c/p\u003e\n\u003cp\u003eM-D: Medium Dose\u003c/p\u003e\n\u003cp\u003eNADPH: Nicotinamide Adenine Dinucleotide Phosphate\u003c/p\u003e\n\u003cp\u003eN-C: Negative Control\u003c/p\u003e\n\u003cp\u003eNCD: Non-Communicable Disease\u003c/p\u003e\n\u003cp\u003eP-C: Positive Control\u003c/p\u003e\n\u003cp\u003erpm: Revolutions Per Minute\u003c/p\u003e\n\u003cp\u003eSD: Sprague-Dawley\u003c/p\u003e\n\u003cp\u003et\u003csub\u003e1/2\u003c/sub\u003e: Elimination Half-life\u003c/p\u003e\n\u003cp\u003eT\u003csub\u003emax\u003c/sub\u003e: Time to Reach Maximum Serum Drug Concentration\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eValavanidis A. Dietary Supplements: Beneficial to Human Health or Just Peace of Mind? A Critical Review on the Issue of Benefit/Risk of Dietary Supplements. 2016.\u003c/li\u003e\n\u003cli\u003ePitetti R, Singh S, Hornyak D, Garcia S, Herr S. Complementary and alternative medicine use in children. Pediatric emergency care. 2001;17(3):165.\u003c/li\u003e\n\u003cli\u003eCott JM. Herb‐Drug Interactions: Theory versus Practice. Molecular nutrition food research. 2008;52(7):745-6.\u003c/li\u003e\n\u003cli\u003eAmeade EPK, Ibrahim M, Ibrahim H-S, Habib RH, Gbedema SY. Concurrent Use of Herbal and Orthodox Medicines among Residents of Tamale, Northern Ghana, Who Patronize Hospitals and Herbal Clinics. Evidence-Based Complementary Aternative Medicine. 2018;2018.\u003c/li\u003e\n\u003cli\u003eOz HS, Chen TS, Nagasawa H. Comparative efficacies of 2 cysteine prodrugs and a glutathione delivery agent in a colitis model. Transl Res. 2007;150(2):122-9.\u003c/li\u003e\n\u003cli\u003eRoberts JC, Charyulu RL, Zera RT, Nagasawa HT. Protection against acetaminophen hepatotoxicity by ribose-cysteine (RibCys). Pharmacol Toxicol. 1992;70(4):281-5.\u003c/li\u003e\n\u003cli\u003eChan WK, Delucchi AB. Resveratrol, a red wine constituent, is a mechanism-based inactivator of cytochrome P450 3A4. Life Sci. 2000;67(25):3103-12.\u003c/li\u003e\n\u003cli\u003eSingh RP, Dhanalakshmi S, Rao AR. Chemomodulatory action of Aloe vera on the profiles of enzymes associated with carcinogen metabolism and antioxidant status regulation in mice. Phytomedicine. 2000;7(3):209-19.\u003c/li\u003e\n\u003cli\u003eNagasawa HT. Method to enhance delivery of glutathione and ATP levels in cells. Google Patents; 2015.\u003c/li\u003e\n\u003cli\u003eNebel A, Schneider BJ, Baker RK, Kroll DJ. Potential metabolic interaction between St. John's wort and theophylline. The Annals of pharmacotherapy. 1999;33(4):502.\u003c/li\u003e\n\u003cli\u003eN\u0026rsquo;guessan BB, Amponsah SK, Dugbartey GJ, Awuah KD, Dotse E, Aning A, et al. In Vitro Antioxidant Potential and Effect of a Glutathione-Enhancer Dietary Supplement on Selected Rat Liver Cytochrome P450 Enzyme Activity. Evidence-Based Complementary and Alternative Medicine. 2018;2018.\u003c/li\u003e\n\u003cli\u003eHorn TL, Reichert MA, Bliss RL, Malejka-Giganti D. Modulations of P450 mRNA in liver and mammary gland and P450 activities and metabolism of estrogen in liver by treatment of rats with indole-3-carbinol. Biochemical pharmacology. 2002;64(3):393-404.\u003c/li\u003e\n\u003cli\u003eSpina E, Pisani F, Perucca E. Clinically significant pharmacokinetic drug interactions with carbamazepine. Clinical pharmacokinetics. 1996;31(3):198-214.\u003c/li\u003e\n\u003cli\u003eNair AB, Jacob S. A simple practice guide for dose conversion between animals and human. Journal of basic and clinical pharmacy. 2016;7(2):27.\u003c/li\u003e\n\u003cli\u003eAppiah-Opong R, de Esch I, Commandeur JN, Andarini M, Vermeulen NP. Structure\u0026ndash;activity relationships for the inhibition of recombinant human cytochromes P450 by curcumin analogues. European journal of medicinal chemistry. 2008;43(8):1621-31.\u003c/li\u003e\n\u003cli\u003eAppiah-Opong R, Commandeur JN, van Vugt-Lussenburg B, Vermeulen NP. Inhibition of human recombinant cytochrome P450s by curcumin and curcumin decomposition products. Toxicology. 2007;235(1-2):83-91.\u003c/li\u003e\n\u003cli\u003eUmegaki K, Saito K, Kubota Y, Sanada H, Yamada K, Shinozuka K. Ginkgo biloba extract markedly induces pentoxyresorufin O-dealkylase activity in rats. The Japanese Journal of Pharmacology. 2002;90(4):345-51.\u003c/li\u003e\n\u003cli\u003eBusby WF, Jr., Ackermann JM, Crespi CL. Effect of methanol, ethanol, dimethyl sulfoxide, and acetonitrile on in vitro activities of cDNA-expressed human cytochromes P-450. Drug metabolism and disposition: the biological fate of chemicals. 1999;27(2):246-9.\u003c/li\u003e\n\u003cli\u003eRiad LE, Chan KK, Sawchuk RJ. Determination of the relative formation and elimination clearance of two major carbamazepine metabolites in humans: a comparison between traditional and pooled sample analysis. Pharmaceutical research. 1991;8(4):541-3.\u003c/li\u003e\n\u003cli\u003ePichard-Garcia L, Hyland R, Baulieu J, Fabre J-M, Milton A, Maurel P. Human hepatocytes in primary culture predict lack of cytochrome P-450 3A4 induction by eletriptan in vivo. Drug metabolism and disposition. 2000;28(1):51-7.\u003c/li\u003e\n\u003cli\u003eBedada SK, Neerati P. Modulation of CYP3A enzyme activity by diosmin and its consequence on carbamazepine pharmacokinetics in rats. Naunyn-Schmiedeberg's archives of pharmacology. 2018;391(2):115-21.\u003c/li\u003e\n\u003cli\u003eFong SYK, Gao Q, Zuo Z. Interaction of carbamazepine with herbs, dietary supplements, and food: a systematic review. Evidence-Based Complementary Alternative Medicine. 2013;2013.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;text-align:justify;line-height:200%;font-size:15px;font-family:\"Calibri\",sans-serif;'\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; line-height: 200%; font-family: \"Times New Roman\", serif; color: rgb(0, 0, 0);'\u003eTable 1:\u003c/span\u003e\u003c/strong\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;line-height:200%;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;Summary of the effect of Cellgevity\u0026reg; on selected rat CYP enzyme activity\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n\u003ctable style=\"border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 125.9pt;border-top: 1pt solid windowtext;border-left: none;border-bottom: 1pt solid windowtext;border-right: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eCYP ISOFORM\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 123.25pt;border-top: 1pt solid windowtext;border-left: none;border-bottom: 1pt solid windowtext;border-right: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eASSAY\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 218.85pt;border-top: 1pt solid windowtext;border-left: none;border-bottom: 1pt solid windowtext;border-right: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eEFFECT OF CELLGEVITY\u0026reg; ON CYP ACTIVITY\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 125.9pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eCYP3A4\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 123.25pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eBROD\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 218.85pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eNo significant increase in enzyme activity\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 125.9pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eCYP2B1/2\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 123.25pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003ePROD\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 218.85pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eNo significant increase in enzyme activity\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 125.9pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eCYP1A1/2\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 123.25pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eMROD\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 218.85pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eSignificant increase in enzyme activity\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e(H-D: p \u0026lt; 0.001)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 125.9pt;border: none;padding: 0in 5.4pt;height: 34.95pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eCYP2C9\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 123.25pt;border: none;padding: 0in 5.4pt;height: 34.95pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eDiclofenac Hydroxylation\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 218.85pt;border: none;padding: 0in 5.4pt;height: 34.95pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eSignificant increase in enzyme activity\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e(L-D: p \u0026lt; 0.05; M-D and H-D: p \u0026lt; 0.001)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 125.9pt;border-top: none;border-right: none;border-left: none;border-image: initial;border-bottom: 1pt solid windowtext;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eCYP2D6\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 123.25pt;border-top: none;border-right: none;border-left: none;border-image: initial;border-bottom: 1pt solid windowtext;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eDextromethorphan\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eO-Demethylation\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 218.85pt;border-top: none;border-right: none;border-left: none;border-image: initial;border-bottom: 1pt solid windowtext;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eSignificant increase in enzyme activity\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e(L-D and M-D: p \u0026lt; 0.05; H-D: p \u0026lt; 0.01)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:200%;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style=\"font-size:16px;line-height:200%;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:150%;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cem\u003e\u003cspan style='font-size:16px;line-height:150%;font-family:\"Times New Roman\",serif;'\u003eL-D = Low dose (38.63 mg/kg) of Cellgevity\u003c/span\u003e\u003c/em\u003e\u003cspan style='font-size:16px;line-height:150%;font-family:\"Times New Roman\",serif;'\u003e\u0026reg;\u003c/span\u003e\u003cem\u003e\u003cspan style='font-size:16px;line-height:150%;font-family:\"Times New Roman\",serif;'\u003e; M-D = Medium dose (77.25 mg/kg) of Cellgevity\u003c/span\u003e\u003c/em\u003e\u003cspan style='font-size:16px;line-height:150%;font-family:\"Times New Roman\",serif;'\u003e\u0026reg;\u003c/span\u003e\u003cem\u003e\u003cspan style='font-size:16px;line-height:150%;font-family:\"Times New Roman\",serif;'\u003e; H-D = High dose (154.50 mg/kg) of Cellgevity\u003c/span\u003e\u003c/em\u003e\u003cspan style='font-size:16px;line-height:150%;font-family:\"Times New Roman\",serif;'\u003e\u0026reg;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;line-height:107%;font-size:15px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n\u003ch1 style='margin:0in;margin-bottom:.0001pt;text-align:justify;line-height:150%;font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003eTable 2: \u003cspan style=\"font-weight:normal;\"\u003ePharmacokinetic parameters of carbamazepine for the two groups of rats administered\u0026nbsp;carbamazepine plus Cellgevity\u0026reg;, or carbamazepine plus normal saline over a period of 14 days.\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/h1\u003e\n\u003ctable style=\"border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 124.55pt;border-top: 1pt solid windowtext;border-left: none;border-bottom: 1pt solid windowtext;border-right: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003ePARAMETERS (UNIT)\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 144.8pt;border-top: 1pt solid windowtext;border-left: none;border-bottom: 1pt solid windowtext;border-right: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003eCARBAMAZEPINE + NORMAL SALINE\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 136.25pt;border-top: 1pt solid windowtext;border-left: none;border-bottom: 1pt solid windowtext;border-right: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003eCARBAMAZEPINE + CELLGEVITY\u0026reg;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 124.55pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003eC\u003csub\u003emax\u0026nbsp;\u003c/sub\u003e(\u0026micro;mol/L)\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 144.8pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e11\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 136.25pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e20\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 124.55pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003eT\u003csub\u003emax\u0026nbsp;\u003c/sub\u003e(h)\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 144.8pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e4\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 136.25pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e4\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 124.55pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003ek\u003csub\u003ee\u0026nbsp;\u003c/sub\u003e(h\u003csup\u003e-1\u003c/sup\u003e)\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 144.8pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e0.41\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 136.25pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e0.28\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 124.55pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003et\u003csub\u003e1/2\u003c/sub\u003e (h)\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 144.8pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e1.7\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 136.25pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e2.3\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 124.55pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003eAUC\u003csub\u003e0\u0026rarr;24\u003c/sub\u003e (\u0026micro;mol.h/L)\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 144.8pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e170\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 136.25pt;border: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e347\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 124.55pt;border-top: none;border-right: none;border-left: none;border-image: initial;border-bottom: 1pt solid windowtext;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align: justify;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cstrong\u003e\u003cspan style='font-size: 16px; font-family: \"Times New Roman\", serif;'\u003eAUC\u003csub\u003e0\u0026rarr; \u0026infin;\u003c/sub\u003e (\u0026micro;mol.h/L)\u003c/span\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 144.8pt;border-top: none;border-right: none;border-left: none;border-image: initial;border-bottom: 1pt solid windowtext;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e178\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 136.25pt;border-top: none;border-right: none;border-left: none;border-image: initial;border-bottom: 1pt solid windowtext;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:.0001pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e375\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;line-height:107%;font-size:15px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style=\"color: rgb(0, 0, 0);\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Carbamazepine, Cellgevity®, Cytochrome P450, Interaction, Pharmacokinetics, Supplement","lastPublishedDoi":"10.21203/rs.2.19774/v2","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.2.19774/v2","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e: There is considerable evidence that many patients concurrently administer dietary supplements with conventional drugs, creating a risk for potential drug-supplement interaction. The aim of this study was to determine the effect of Cellgevity® supplement on selected rat liver cytochrome P450 (CYP) enzymes. \u003cstrong\u003eAlso, we sought to deternine the effect of Cellgevity® on the pharmacokinetics of carbamazepine, a CYP3A4 substrate. \u003c/strong\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eMethods: \u003c/strong\u003eMale\u003cstrong\u003e \u003c/strong\u003eSprague-Dawley (SD) rats were randomly put into 5 groups and administered (\u003cem\u003eper os\u003c/em\u003e) either distilled water (negative control), Cellgevity® (3 separate doses) or phenobarbital (positive control). Modulation of liver CYP enzyme activity was evaluated after 30 days of treatment, \u003cstrong\u003eusing probe substrates, spectroscopic and high-performance liquid chromatographic methods. \u003c/strong\u003eIn the pharmacokinetic study, 12 SD rats were divided into 2 groups administered (\u003cem\u003eper os\u003c/em\u003e) carbamazepine plus Cellgevity®, or carbamazepine plus normal saline, both over a period of 14 days. \u003cstrong\u003eBlood samples from rats in the same group were collected after treatment. Serum samples\u0026nbsp;were prepared and pooled together at each specific sampling time point. Levels of carbamazepine were determined using a fluorescence polarization immunoassay.\u003c/strong\u003e \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eActivities of\u003cstrong\u003e \u003c/strong\u003erat liver\u003cstrong\u003e \u003c/strong\u003eCYP1A1/2, CYP2C9 and CYP2D6 were significantly increased by Cellgevity® after 30-day treament. Pharmacokinetic parameters for rats administered carbamazepine with Cellgevity® vis-a-vis carbamazepine with normal saline were as follows: C\u003csub\u003emax\u003c/sub\u003e; 20 μmol/L vs 11 μmol/L, AUC\u003csub\u003e0→24\u003c/sub\u003e; 347 μmol.h/L vs 170 μmol.h/L, K\u003csub\u003ee\u003c/sub\u003e; 0.28 h\u003csup\u003e-1\u003c/sup\u003e vs 0.41 h\u003csup\u003e-1\u003c/sup\u003e, and t\u003csub\u003e1/2\u003c/sub\u003e; 2.3 h vs 1.7 h, respectively.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusions:\u003c/strong\u003e Cellgevity® increased the activity of rat CYP1A1/2, CYP2C9 and CYP2D6 enzymes, and altered the pharmacokinetics of carbamazepine in rats.\u003c/p\u003e","manuscriptTitle":"Effect of Cellgevity® Supplement on Selected Rat Liver Cytochrome P450 Enzyme Activity and Pharmacokinetic Parameters of Carbamazepine","msid":"","msnumber":"","nonDraftVersions":[{"code":2,"date":"2020-04-20 15:02:48","doi":"10.21203/rs.2.19774/v2","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}},{"code":1,"date":"2019-12-31 17:14:11","doi":"10.21203/rs.2.19774/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"275faded-edb6-4557-8474-b9a29c088653","owner":[],"postedDate":"April 20th, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":45925,"name":"Clinical Pharmacology"}],"tags":[],"updatedAt":"2020-05-29T02:57:52+00:00","versionOfRecord":[],"versionCreatedAt":"2020-04-20 15:02:48","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v2","identity":"rs-10386","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"identity":"rs-10386","version":["v2"]},"buildId":"FbvkV6FR0MCFSLy54lSbu","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}