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Advances in Clinical Toxicology Research Article 19 min read

Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Extract

Lin JY and Chiu E*
* Corresponding author
ISSN: 2577-4328  10.23880/act-16000146  Received: February 12, 2019  Published: February 23, 2019
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 19 references
 3 figures
 6 tables
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Keywords
Taiwanofungus camphoratus hERG Central Nervous System Respiratory System Cardiovascular System
Abstract

Taiwanofungus camphoratus is a unique mushroom that only grows in Taiwan. It has been used as fold medicine for a long history. Recent studies have demonstrated T. camphoratus possessed multiple pharmacological effects including anti-cancer, hepatoprotective and immunomodulatory effects. T. camphoratus extract was composed of extracts from cutlog cultivated fruiting body and solid-state culture of T. camphoratus. This article presents the testing results of T. camphoratus extract in in vitro hERG assay and in vivo safety pharmacology studies on central nervous, respiratory and cardiovascular systems. Results 1) The hERG transfected HEK293 cells were treated with T. camphoratus extract at concentrations of 5, 10 and 25 µg/mL showed no significant effect on hERG current. 2) T. camphoratus extract was found to have no significant effects on central nervous and respiratory systems of male rats and female rats at oral doses up to 3400 mg/kg and 1700 mg/kg, respectively. 3) Beagle dogs received T. camphoratus extract orally up to the dosage of 1000 mg/kg did not cause physiological abnormalities on cardiovascular system. Accordingly, these results provided the safety information of T. camphoratus extract for human consumption.

Introduction

Taiwanofungus camphoratus (syn. Antrodia cinnamomea, Antrodia camphorata) is an edible and medicinal mushroom originating in Taiwan. Taiwan aborigines have commonly used the fruiting body of T. camphoratus as folk medicine for health promotion and treating liver disease, drug and food intoxication, hypertension and cancer [1, 2]. Many pharmacologic studies have noted that T. camphoratus possessed a variety of biological activities including anti-oxidant, anti- cancer, liver protection, anti-inflammation, and immunomodulatory effects [3, 4, 5, 6, 7, 8, 9, 10, 11]. Additionally, many bioactive components of T. camphoratus have been identified, including terpenoids, polysaccharides, benzenoids, lignans, nucleic acid, benzoquinone derivatives, steroids, and maleic/succinic acid derivatives [1, 12]. For the demand of market, it has been developed many kinds of cultivation methods to produce T. camphoratus including liquid fermentation, solid-state culture, cut wood culture, and dish culture. The components of T. camphoratus will depend on the culture techniques.

With the wild applications of T. camphoratus used in health food supplements in Taiwan, the safety issue of T. camphoratus has become increasingly important to consumers. Several toxicological studies have been done to support the safety of T. camphoratus. In 2013, Chang et al [13], reported that no abnormal findings were observed in male and female mice up to 1666.67 mg/kg. Huang et al [14], demonstrated that under the dosage of 6 g/kg of T. camphoratus showed no sub-chronic toxicity and teratogenicity in SD rats. Our previous studies also found the health food product “Leader Deluxe Antrodia cinnamomea” and “Leader Antrodia cinnamomea capsule” have no obvious toxic evidences in rats at dose of 2800 and 2500 mg/kg, respectively [15, 16]. The maximum tolerated dose (MTD) of the solid-state cultivated mycelial powder of Antrodia cinnamomea (LE-SC) was greater than 13.3 g/kg bw and 90 days repeated dose oral toxicity studies also showed no significant toxicity signs in both male and female rats up to the dose of 7.6 g/kg bw [17].

T. camphoratus extract was composed of extract from cut-log cultivated fruiting body and solid-state culture of T. camphoratus. The previous studies from our laboratory revealed T. camphoratus extract showed no toxicity evidences at dose of 1700 mg/kg in 90 and 180 days repeated oral dose toxicity studies in rats (data not published). In this article, we evaluated the effects of T. camphoratus extract in in vitro primary cardiovascular test (hERG test) and in vivo core batteries of safety pharmacology studies on central nervous, respiratory and cardiovascular systems. All studies were designed according to the suggestions of “ICH (2001) S7A Safety Pharmacology Studies for Human Pharmaceuticals”. This is the first study to explore the safety pharmacology effects of T. camphoratus and the results would provide more safety evidences for T. camphoratus.

Material and Methods

Test Substance

T. camphoratus extract, LEAC-102, was composed of extracts from cut-log cultivated fruiting body and solid- state culture of T. camphoratus that provided by Taiwan Leader Biotech Corp. (Taipei, Taiwan).

Lin JY and Chiu E. Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Camphoratus Extract. Adv Clin Toxicol 2019, 4(1): 000146.

Evaluation on hERG Channel Current

Cell culture: Human ether-a-go-go-related gene transfected human embryonic kidney 293 cells (hERG transfected HEK293 Cells) were obtained from the University of Wisconsin (Madison, WI). The cells were maintained and passaged in Minimum Essential Medium (Sigma-Aldrich, St. Louis, MO) supplemented with 10 % heat-inactivated fetal bovine serum, 100 units/mL penicillin-streptomycin, 1 mmol/L sodium pyruvate, MEM non-essential amino acids (all supplements were from Life Technologies Corporation, Carlsbad, CA) and 400 μg/mL G 418 (Geneticin; Sigma-Aldrich, St. Louis, MO). The cells were cultured in a 5% CO2 incubator at 37°C. Electrophysiological experiments: The tests were conducted with the whole-cell patch-clamp method at physiological temperature (37±1.0°C). T. camphoratus extract was dissolved in dimethylfloxide (DMSO) and subsequently prepared by diluting the DMSO solutions with the superfusing solution (137 mmol/L NaCl, 4 mmol/L KCl, 1.8 mmol/L CaCl2, 1 mmol/L MgCl2, 10 mmol/L HEPES, and 10 mmol/L D(+)-glucose, pH adjusted to 7.4 with NaOH) to obtain the test solutions of 5, 10, and 25 µg/mL. The test solutions were filtered through a membrane filter (0.5 μm; PTFE Hydrophilic membrane; Advantec Tokyo Kaisha, Ltd.) to remove fibril- like forms undissolved in the superfusing solution. The resultant solutions were used for the hERG-current measurement. The peak amplitude of the hERG tail currents was measured from 4 individual cells, which were assigned to each experimental group including test solutions at the respective concentrations (5, 10, and 25 µg/mL), 0.5 vol% DMSO (vehicle control) or 0.1 µmol/L E- 4031 (positive control). The pipette solution was composed of 130 mmol/L KCl, 1 mmol/L MgCl2, 5 mmol/L EGTA, 10 mmol/L HEPES, and 5 mmol/L MgATP (pH adjusted to 7.2 with KOH).

The hERG currents passing through the cell membrane were measured under voltage clamp mode by the whole- cell patch-clamp technique. A schematic diagram of the voltage protocol to elicit the hERG tail current is shown below (Figure 1); the membrane potential of the cell was held at −80 mV, and depolarizing step pulses were given every 15 seconds to elicit the hERG tail current. The effects of the vehicle control, test substance, or positive- control substance on the hERG current were determined by changes in the peak amplitude of the tail current elicited by a partially repolarizing step pulse from +20 mV to −50 mV for 500 milliseconds following a depolarizing step pulse from the holding potential of −80 mV to +20 mV for 500 milliseconds. The peak value of the tail current was computed based on the holding current. After confirming a stable baseline for the peak tail currents, the test solution was applied to the cell for 11 mins at a flow rate of 5 mL/min with a peristaltic pump (WM-120S/DV; Watson-Marlow Limited, Falmouth, UK).

Figure 1: hERG Tail Current. Data acquisition and analysis: The hERG currents were measured with an amplifier (Axopatch 200B; Molecular Devices, LLC., Sunnyvale, CA). Electric signals were recorded onto computer hard drive by software (pCLAMP 10; Molecular Devices, LLC., Sunnyvale, CA). The peak tail currents obtained before and 11 mins after beginning the application were compared, and the change rate (suppression rate) was calculated. The suppression rate in each cell was compensated for by the mean suppression rate in the vehicle-control group with the formula described below. Effects of the test substance and positive-control substance were evaluated with the compensated suppression rates.  X: Suppression rate (%) X = [(A – B) / A] × 100 A: Peak tail current in each cell immediately before application B: Peak tail current in each cell at completion of application  Xc: Compensated suppression rate (%) Xc = [(A − B) / (100 − B)] × 100 A: Suppression rate in each cell (%) B: Mean suppression rate in vehicle-control group (%) The data are represented as mean ± standard deviation (SD) and analyzed using one-way analysis of variance (one-way ANOVA) or student’s t-test (SAS®, Ver. 9.3; SAS Institute Japan Ltd. and EXSUS, Ver. 8.0; CAC Croit Corporation). The significance levels were defined at _p_<0.05.
Click to enlarge
Figure 1: hERG Tail Current. Data acquisition and analysis: The hERG currents were measured with an amplifier (Axopatch 200B; Molecular Devices, LLC., Sunnyvale, CA). Electric signals were recorded onto computer hard drive by software (pCLAMP 10; Molecular Devices, LLC., Sunnyvale, CA). The peak tail currents obtained before and 11 mins after beginning the application were compared, and the change rate (suppression rate) was calculated. The suppression rate in each cell was compensated for by the mean suppression rate in the vehicle-control group with the formula described below. Effects of the test substance and positive-control substance were evaluated with the compensated suppression rates.  X: Suppression rate (%) X = [(A – B) / A] × 100 A: Peak tail current in each cell immediately before application B: Peak tail current in each cell at completion of application  Xc: Compensated suppression rate (%) Xc = [(A − B) / (100 − B)] × 100 A: Suppression rate in each cell (%) B: Mean suppression rate in vehicle-control group (%) The data are represented as mean ± standard deviation (SD) and analyzed using one-way analysis of variance (one-way ANOVA) or student’s t-test (SAS®, Ver. 9.3; SAS Institute Japan Ltd. and EXSUS, Ver. 8.0; CAC Croit Corporation). The significance levels were defined at p<0.05.

Figure 1: hERG Tail Current. Data acquisition and analysis: The hERG currents were measured with an amplifier (Axopatch 200B; Molecular Devices, LLC., Sunnyvale, CA). Electric signals were recorded onto computer hard drive by software (pCLAMP 10; Molecular Devices, LLC., Sunnyvale, CA). The peak tail currents obtained before and 11 mins after beginning the application were compared, and the change rate (suppression rate) was calculated. The suppression rate in each cell was compensated for by the mean suppression rate in the vehicle-control group with the formula described below. Effects of the test substance and positive-control substance were evaluated with the compensated suppression rates.  X: Suppression rate (%) X = [(A – B) / A] × 100 A: Peak tail current in each cell immediately before application B: Peak tail current in each cell at completion of application  Xc: Compensated suppression rate (%) Xc = [(A − B) / (100 − B)] × 100 A: Suppression rate in each cell (%) B: Mean suppression rate in vehicle-control group (%) The data are represented as mean ± standard deviation (SD) and analyzed using one-way analysis of variance (one-way ANOVA) or student’s t-test (SAS®, Ver. 9.3; SAS Institute Japan Ltd. and EXSUS, Ver. 8.0; CAC Croit Corporation). The significance levels were defined at p<0.05.

Lin JY and Chiu E. Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Camphoratus Extract. Adv Clin Toxicol 2019, 4(1): 000146.

Evaluation of Central Nervous System (CNS) in Rats

The 8 weeks old Sprague Dawley (SD) (BioLASCO, Taiwan Co. Ltd.) rats were randomly divided into four groups with 10 males and 10 females in each group. The animals were housed in the AAALAC International accredited facility of Level Biotech. Inc. under 12 h light/12h dark cycle and the temperature of animal room was at 19.7-20.0℃ with relative humidity 45.9-63.0 %. The test article, T. camphoratus extract, was suspended in water for injection (WFI) to obtain dosing solutions. The dosing solutions were orally administered to animals at dose of 0, 170, 1700 and 3400 mg/kg for male rats and 0, 170, 850 and 1700 mg/kg for female rats. The control group was administrated with WFI only. The motor activity was conducted on all animals at pre-dose, 30 ± 5 mins post-dose and 24 ± 2 h post-dose and the functional observation battery (FOB) was performed at pre-dose, 1.5 ± 0.5 h post-dose and 24 ± 2 h post-dose. The FOB was performed of all animals including cage-side observation and handling observation (the testing parameters were including lethality, convulsion, tremor, straub tail, sedation, excitation, abnormal gait, jumps, loss of balance, motor incoordination, fore-paw treading, abnormal writhes, piloerection, stereotypies, head twitches, scratching, respiration, fear, touch response, sedation intensity, excitation intensity, aggressiveness, right reflex, ptosis, exophthalmia, grip strength, akinesia, catalepsy, corneal reflex, analgesia, defecation, salivation, lacrimation, pupillary light reflex and body temperature). All data were calculated and expressed as mean ±SD or percentage. Comparisons of parametric data collected from treated and control groups were performed by one- way ANOVA, followed by Dunnett’s method (SPSS, Ver. 12.0). Non-parametric data was analyzed by Kruskal- Wallis nonparametric ANOVA method. The significance level was defined at p<0.05. The study was approved by the Institutional Animal Care and Use Committee (IACUC number: 170206-02).

Evaluation of Respiratory System in Rats

The 8-9 weeks old SD (BioLASCO, Taiwan Co. Ltd.) rats were randomly divided into four groups with 10 males and 10 females in each group. The animals were also housed in Level Biotech. Inc. under 12 h light/12h dark cycle and the temperature of animal room was at 19.7- 20.0℃ with relative humidity 45.7-61.9 %. The test article, T. camphoratus extract, was suspended in WFI to obtain dosing solutions. The dosing solutions were orally administered to animals at dose of 0, 170, 1700 and 3400 mg/kg for male rats and 0, 170, 850 and 1700 mg/kg for female rats. The control group was administrated with WFI only. The respiratory parameters were detected on all animals before study for 1 h recording as baseline, 4 h recording continuously after dosing and the 24 to 25 h recording after dosing. The whole-body plethysmography (emka TECHNOLOGIES, Paris, France) was used for detecting the respiratory parameters in freely moving animals. The parameters included inspiratory time, expiration time, peak inspiratory flow, peak expiratory flow, tidal volume, expired volume, relaxation time, minute volume, frequency of breathing, end-inspiratory pause, end-expiratory pause, enhanced pause and mid- expiratory flow. All study data acquisition and analysis were operated under iox software system (emka TECHNOLOGIES, Paris, France) and an average value of each selected parameter was calculated from detectable peaks at 1 min interval. The data for each respiratory parameter was calculated and presented as baseline value (one hour recording data prior to dosing), four hours continuous recording data at one hour interval after dosing and 24 to 25 h recording after dosing (denoted as the 24th h time point). All data were expressed as mean ± SD and analyzed by one-way ANOVA, followed by Dunnett’s method (SPSS, Ver. 12.0). Besides, an additional paired t-test was used if the ANOVA results were statistically significant. The significance level was defined at p<0.05. The study was approved by the Institutional Animal Care and Use Committee (IACUC number: 180101).

Evaluation of Cardiovascular System in Beagle dogs

The 6 months old beagle dogs (Covance Inc., Cumberland, VA) were housed in the AAALAC International accredited facility of Level Biotech. Inc. under 12 h light/12h dark cycle and the temperature of animal room was at 19.2-22.1℃ with relative humidity 43.7-67.2%. Total six beagle dogs (3 male and 3 female) were used and treated with vehicle control (Empty Porcine Hard Gelatin Capsules) first and following treated with each dosage of T. camphoratus extract (54, 540, 1000 mg/kg) in gelatin capsules with at least 1-week washout period between treatments. The emkaPACK_4G_ noninvasive telemetry system (emka TECHNOLOGIES, Paris, France) was used for detecting the cardiovascular parameters in freely moving animals. The parameters included RR interval, PR interval, P wave duration, QRS wave interval, QT interval, QTcB (Bazett's method), QTcF (Fridericia's method), heart rate, diastolic arterial pressure, systolic arterial pressure and mean arterial Lin JY and Chiu E. Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Camphoratus Extract. Adv Clin Toxicol 2019, 4(1): 000146.

pressure. The electrocardiograms parameters were detected on all animals during pre-dose period for 1 h recording as baseline, and 24 h recording continuously after each dosage. The tail artery blood pressure was measured (ecgAUTO software NIBP) during the pre-dose period, 0-4th and 23th-24th h post-dosing periods. All study data acquisition was operated under iox software system (emka TECHNOLOGIES, Ver. 2.9.4.25). Study data analysis was operated under ecgAUTO software system (emka TECHNOLOGIES, Ver. 3.3.0.21). All ECG parameters were extracted from the lead II configuration. All data were expressed as mean ± SD and analyzed by one-way ANOVA, followed by Dunnett’s method (SPSS, Ver. 12.0). Besides, an additional paired t-test was used if the ANOVA results were statistically significant. The significance level was defined at p<0.05. The study was approved by the Institutional Animal Care and Use Committee (IACUC number: 171106).

Results and Discussion

Effects of T. camphoratus Extract on hERG Current in HEK293 Cells

The effects of the soluble fraction of T. camphoratus extract on hERG current were shown in Figures 2 & 3. The hERG-current-suppression rates, compensated for by the vehicle-control group rate (11.9%), were 2.6%, 4.7%, and 5.2% at dose of 5, 10, and 25 μg/mL, respectively. There was no statistically significant difference between T. camphoratus extract groups and vehicle-control group. A hERG channel inhibitor, E-4031, was used at 0.1 μmol/L as a positive-control substance, and its hERG-current- suppression rate, compensated for by the mean suppression rate in the vehicle-control group, was 86.3%. This rate was statistically significant when compared to the vehicle-control group, thereby confirming the validity of this experimental system to evaluate the suppressive effects of the test substance on the hERG current. However, T. camphoratus extract was a kind of herb material and couldn’t completely dissolve in DMSO solution. The test solutions were filtered through a 0.5μm PTFE Hydrophilic membrane to remove a few fibril-like forms for prevention of interference the experiments. Overall, T. camphoratus extract had no significant effect on the hERG current at nominal concentrations of up to 25μg/mL, which were based on the actual weight of the test substance in preparation of the DMSO solution, under the conditions of this study.

Figure 2: Representative hERG Current Waveforms.
Click to enlarge
Figure 2: Representative hERG Current Waveforms.

Vehicle control: 0.5-vol% DMSO; Positive control: 0.1 μmol/L E-4031.

Figure 3: Effects of T. Camphoratus Extract on Tail Current from hERG Channels Expressed in HEK293 Cells.
Click to enlarge
Figure 3: Effects of T. Camphoratus Extract on Tail Current from hERG Channels Expressed in HEK293 Cells.

Lin JY and Chiu E. Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Camphoratus Extract. Adv Clin Toxicol 2019, 4(1): 000146.

No significant suppressive effect of T. camphoratus extract on the hERG current. The mean suppression rate compensated for by 11.9%, the mean suppression rate in the vehicle-control group (0.5-vol% DMSO), 11 minutes after application was as follows: 2.6% ± 4.4% at 5 μg/mL, 4.7% ± 4.0% at 10 μg/mL, and 5.2% ± 5.9% at 25 μg/mL. The mean compensated suppression rate in the positive- control group (0.1 μmol/L E-4031) was 86.3% ± 3.5%. Each column represents the mean ± SD (n = 4). No statistically significant difference was noted among the test substance and vehicle-control groups, one-way ANOVA, ††p<0.01, Student’s t-test.

The male and female rats received T. camphoratus extract at doses up to 3400 mg/kg and 1700 mg/kg, respectively, showed no significant effects on CNS. Results of motor activity and functional observational battery (FOB) responses were shown in Tables 1 & 2. No treatment related changes were observed in motor activity at pre-dose period, 30 mins and 24 h post-dosing periods. In FOB measurements, the value of grip strength in high-dose (3400 mg/kg) males was statistically significantly higher than vehicle control group at 24 h post-dosing period. However, this change was considered incidental and unrelated to treatment because the change was not correlated with other parameters of motor activity and FOB. Based on the results, T. camphoratus extract showed no adverse effects on CNS in rats that could provide safety information for human exposure.

Effects of T. camphoratus Extract on Central Nervous System (CNS) in Rats

ParametersGenderDose (mg/kg)Pre-dose30 min24 h
Average speed (cm/sec.)
Male05.424±0.9664.183±1.0344.203±1.355
1705.315±0.5974.271±0.8164.099±1.161
17005.502±0.8984.305±0.6444.797±0.741
34005.338±1.0124.973±0.6824.230±0.772
Female05.430±0.6244.113±0.9524.504±0.895
1705.382±1.7204.056±1.2564.824±1.265
8505.478±1.3284.231±1.1044.615±1.346
17005.288±1.6414.198±0.8505.256±1.402
Max speed (cm/sec.)
Male041.996±10.90434.101±11.97127.244±12.993
17040.515±7.52131.513±5.76028.439±12.583
170042.572±15.21033.105±7.39537.382±8.473
340046.482±8.68741.703±10.61138.373±11.421
Female051.748±8.95440.166±17.73743.893±18.941
17045.023±16.18535.371±22.63440.370±17.980
85046.491±8.77737.963±15.30245.788±15.866
170059.697±34.94048.554±19.33755.333±8.855
Total distance (cm)
Male03270.646±1149.9002458.525±1023.3022642.129±1344.129
1703346.333±722.5972657.786±712.8402840.399±1169.978
17003967.793±1340.8632854.312±1062.8883574.599±877.609
34003862.154±1466.5862974.510±1011.1232900.317±1037.412
Female02994.735±712.0932152.536±658.9272767.476±1208.073
1703689.441±1805.1362503.654±740.3673344.445±1093.070
8503734.458±2002.9422376.107±1022.1693343.206±1081.230
17003521.432±1479.4362502.781±645.5903649.590±1210.729

Table 1: Effect of T. Camphoratus Extract on Motor Activity in Rats. All data presented as mean ± SD.

Lin JY and Chiu E. Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Camphoratus Extract. Adv Clin Toxicol 2019, 4(1): 000146.

ParametersGenderDose (mg/kg)Pre-dose1.5 h24 h
Respiration (breath per min.)
Male0126.6±14.0106.8±7.4111.6±9.9
170125.4±10.8115.8±8.0117.6±9.5
1700120.6±7.7110.4±11.7117.0±12.1
3400122.4±13.6112.8±6.8112.2±13.6
Female0114.6±11.196.0±16.294.2±9.8
170119.4±16.196.6±10.088.2±13.9
850105.6±12.199.6±13.093.0±11.7
1700108.0±12.097.8±12.792.4±11.7
Grip strength (kg)
Male00.63±0.070.69±0.060.66±0.07
1700.69±0.070.66±0.110.68±0.04
17000.63±0.080.69±0.060.69±0.06
34000.66±0.070.68±0.060.74±0.05 *
Female00.60±0.070.65±0.080.65±0.07
1700.59±0.060.62±0.060.69±0.06
8500.62±0.060.66±0.050.68±0.04
17000.58±0.060.64±0.080.68±0.08
Defecation
Male00.5±0.51.0±0.00.7±0.5
1700.8±0.40.7±0.50.4±0.5
17000.5±0.51.0±0.00.4±0.5
34000.5±0.50.6±0.50.5±0.5
Female00.7±0.50.8±0.41.0±0.0
1700.9±0.30.9±0.30.8±0.4
8500.9±0.30.7±0.51.0±0.0
17000.9±0.30.9±0.30.9±0.3
Body temperature Scores
Male0-0.1±0.30.0±0.00.0±0.0
1700.0±0.00.0±0.00.0±0.0
17000.0±0.00.0±0.50.0±0.0
3400-0.1±0.30.0±0.00.0±0.0
Female00.0±0.00.0±0.0-0.2±0.4
1700.0±0.00.0±0.0-0.2±0.4
8500.0±0.00.0±0.00.0±0.0
17000.0±0.00.0±0.0-0.1±0.3

Table 2: Effect of T. Camphoratus Extract on Functional Observation Battery in Rats. All data presented as mean ± SD. * p< 0.05 c

Effects of T. camphoratus Extract on Respiratory System in Rats

The male and female rats received T. camphoratus extract at doses up to 3400 mg/kg and 1700 mg/kg, respectively, showed no significant effects on respiratory system. There was no statistical difference noted in all respiratory parameters among all treated groups during Lin JY and Chiu E. Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Camphoratus Extract. Adv Clin Toxicol 2019, 4(1): 000146.

each time period (Table 3). Based on the test results, T. camphoratus extract administered to rats via oral gavage up to the dosage of 3400 mg/kg for males and 1700 mg/kg for females did not cause significant abnormal respiratory effects in this study. All results generated from this study will provide safety information for human exposure.

ParametersGender Dose (mg/kg) Pre-dose 1 h 2 h 3 h 4 h 24 h Inspiratory Time (msec) Male 0 200.34 ± 25.95 169.74 ± 27.42 218.75 ± 24.24 216.94 ± 14.52 219.42 ± 22.57 201.12 ± 26.08 170 216.34 ± 22.56 169.43 ± 40.69 209.00 ± 33.81 214.32 ± 20.86 211.94 ± 36.31 217.51 ± 19.92 1700 206.80 ± 17.05 179.65 ± 27.40 209.31 ± 27.36 212.86 ± 27.88 210.93 ± 34.55 206.79 ± 18.14 3400 209.92 ± 26.24 161.14 ± 28.06 210.60 ± 24.84 225.75 ± 20.21 220.75 ± 26.18 215.81 ± 27.27 Female 0 187.38 ± 58.44 178.32 ± 27.67 260.15 ± 28.83 248.82 ± 22.91 254.34 ± 36.85 240.97 ± 36.48 170 161.36 ± 44.39 165.94 ± 36.96 232.59 ± 32.92 259.87 ± 37.36 262.10 ± 24.54 230.52 ± 51.11 850 188.80 ± 40.22 175.45 ± 34.88 241.26 ± 45.74 250.23 ± 30.97 255.69 ± 22.14 238.89 ± 23.05 1700 189.44 ± 42.26 207.44 ± 40.58 248.24 ± 25.60 235.01 ± 59.60 244.48 ± 47.57 249.50 ± 29.30 Expiration Time (msec) Male 0 284.01 ± 37.39 264.67 ± 36.04 313.36 ± 37.68 314.75 ± 33.81 324.63 ± 42.57 295.30 ± 34.41 170 290.18 ± 31.09 248.84 ± 46.29 298.65 ± 33.30 305.01 ± 36.65 303.86 ± 50.67 305.75 ± 29.16 1700 274.96 ± 25.10 262.90 ± 40.88 294.78 ± 33.96 295.36 ± 35.08 299.09 ± 33.90 289.19 ± 26.32 3400 286.84 ± 54.55 252.51 ± 46.63 294.48 ± 43.32 306.74 ± 46.06 306.30 ± 45.20 299.14 ± 50.31 Female 0 329.39 ± 97.33 323.53 ± 68.06 456.26 ± 78.57 421.07 ± 44.61 436.07 ± 81.17 413.01 ± 73.20 170 274.69 ± 78.53 291.07 ± 51.51 410.96 ± 80.30 437.24 ± 72.00 449.93 ± 77.51 386.97 ± 93.80 850 296.45 ± 61.10 300.88 ± 52.91 394.30 ± 52.12 397.16 ± 44.19 407.98 ± 44.11 392.59 ± 31.93 1700 301.30 ± 65.78 383.85 ± 134.95431.83 ± 123.98 379.18 ± 86.56 386.96 ± 68.95 398.67 ± 46.46 Peak Inspiratory Flow (mL/s) Male 0 10.93 ± 2.90 13.53 ± 2.36 8.22 ± 1.98 8.66 ± 1.41 8.51 ± 1.63 11.07 ± 3.01 170 8.93 ± 2.29 13.86 ± 4.77 9.35 ± 3.57 8.42 ± 2.28 9.56 ± 4.31 9.35 ± 2.61 1700 9.68 ± 1.41 12.67 ± 1.88 9.83 ± 2.18 9.28 ± 2.89 9.55 ± 2.36 9.93 ± 1.01 3400 9.26 ± 1.79 13.99 ± 2.21 9.04 ± 2.05 7.66 ± 1.39 7.79 ± 1.63 10.11 ± 4.83 Female 0 13.21 ± 3.92 13.17 ± 2.24 7.07 ± 1.25 7.84 ± 1.52 7.37 ± 2.02 8.00 ± 2.44 170 15.38 ± 6.75 14.03 ± 6.09 9.34 ± 2.72 7.24 ± 2.01 7.74 ± 4.71 9.03 ± 3.54 850 13.13 ± 3.56 12.74 ± 3.34 8.35 ± 3.51 7.87 ± 2.34 7.29 ± 1.58 8.19 ± 1.50 1700 12.73 ± 3.39 10.71 ± 3.23 7.36 ± 2.02 9.30 ± 3.64 8.28 ± 2.66 7.04 ± 1.65 Peak Expiratory Flow (mL/s) Male 0 11.59 ± 2.12 12.39 ± 1.00 9.61 ± 1.66 9.83 ± 1.59 9.97 ± 1.49 11.93 ± 2.78 170 10.42 ± 2.20 12.77 ± 3.37 10.39 ± 2.39 9.73 ± 2.10 10.68 ± 2.87 11.07 ± 3.05 1700 10.70 ± 1.43 11.64 ± 1.63 10.45 ± 1.60 10.18 ± 1.72 10.31 ± 1.35 10.99 ± 1.51 3400 10.56 ± 1.33 12.35 ± 1.52 9.83 ± 1.14 9.22 ± 0.89 9.35 ± 1.08 11.41 ± 3.89 Female 0 11.81 ± 2.60 11.28 ± 1.86 7.77 ± 0.64 8.12 ± 1.22 7.83 ± 1.47 8.61 ± 1.33 170 14.30 ± 6.11 12.50 ± 5.96 9.69 ± 2.74 8.03 ± 2.19 8.32 ± 4.01 9.26 ± 2.03 850 12.81 ± 3.14 11.05 ± 2.28 8.77 ± 1.81 8.35 ± 1.77 8.10 ± 1.31 8.85 ± 1.09 1700 12.31 ± 2.81 9.81 ± 1.89 7.84 ± 1.92 9.28 ± 2.46 8.40 ± 1.33 7.81 ± 1.40 Tidal Volume (mL) Male 0 1.12 ± 0.20 1.17 ± 0.18 1.02 ± 0.17 1.04 ± 0.14 1.03 ± 0.13 1.20 ± 0.23 170 1.03 ± 0.23 1.14 ± 0.21 1.05 ± 0.18 1.01 ± 0.21 1.04 ± 0.21 1.12 ± 0.22 1700 1.11 ± 0.09 1.22 ± 0.15 1.08 ± 0.14 1.05 ± 0.11 1.09 ± 0.12 1.16 ± 0.18 3400 1.10 ± 0.08 1.16 ± 0.12 1.01 ± 0.11 1.00 ± 0.09 1.00 ± 0.09 1.17 ± 0.28 Female 0 1.00 ± 0.05 0.99 ± 0.06 0.87 ± 0.07 0.89 ± 0.10 0.87 ± 0.07 0.92 ± 0.10 170 1.02 ± 0.31 0.98 ± 0.30 0.98 ± 0.29 0.88 ± 0.24 0.90 ± 0.28 0.92 ± 0.11 850 1.04 ± 0.10 1.05 ± 0.12 0.91 ± 0.12 0.92 ± 0.09 0.91 ± 0.07 0.93 ± 0.08 1700 0.98 ± 0.12 0.95 ± 0.14 0.87 ± 0.16 0.93 ± 0.12 0.91 ± 0.10 0.87 ± 0.11 Expired Volume (mL) Lin JY and Chiu E. Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Camphoratus Extract. Adv Clin Toxicol 2019, 4(1): 000146.

Male 0 1.12 ± 0.20 1.16 ± 0.17 1.02 ± 0.17 1.05 ± 0.14 1.02 ± 0.14 1.19 ± 0.23 170 1.04 ± 0.25 1.14 ± 0.21 1.05 ± 0.18 1.02 ± 0.21 1.03 ± 0.20 1.14 ± 0.24 1700 1.13 ± 0.09 1.22 ± 0.12 1.10 ± 0.12 1.05 ± 0.11 1.09 ± 0.13 1.17 ± 0.21

34001.10 ± 0.081.18 ± 0.091.01 ± 0.111.01 ± 0.090.99 ± 0.091.17 ± 0.28
Female01.00 ± 0.051.00 ± 0.070.87 ± 0.070.88 ± 0.110.89 ± 0.060.91 ± 0.11
1701.03 ± 0.310.98 ± 0.300.96 ± 0.270.89 ± 0.250.91 ± 0.280.92 ± 0.11
8501.05 ± 0.081.05 ± 0.120.90 ± 0.120.92 ± 0.090.92 ± 0.060.93 ± 0.08
17000.99 ± 0.130.96 ± 0.130.86 ± 0.180.96 ± 0.100.95 ± 0.100.88 ± 0.11
Relaxation Time (msec)
Male0122.26 ± 11.22119.61 ± 10.02129.47 ± 13.59131.90 ± 14.28132.31 ± 14.27127.24 ± 12.34
170123.81 ± 11.61116.87 ± 16.00127.44 ± 11.17128.19 ± 10.80125.30 ± 15.99128.15 ± 11.72
1700123.53 ± 7.60124.82 ± 18.40130.09 ± 11.75129.98 ± 13.69131.30 ± 13.03126.67 ± 8.35
3400123.74 ± 11.39118.98 ± 12.42126.23 ± 9.39131.61 ± 9.46130.01 ± 11.80126.89 ± 12.30
Female0132.33 ± 27.18138.18 ± 28.97177.75 ± 40.53170.53 ± 31.60173.17 ± 35.63155.82 ± 23.36
170113.63 ± 25.11127.74 ± 27.73165.54 ± 55.50179.34 ± 62.47184.04 ± 60.26142.38 ± 18.61
850123.56 ± 19.09133.79 ± 17.38152.94 ± 18.85158.39 ± 17.18160.81 ± 15.11148.80 ± 8.23
1700122.02 ± 18.37156.85 ± 52.22189.01 ± 90.36167.60 ± 56.79161.13 ± 26.39152.71 ± 16.32
Minute Volume (mL)
Male0168.58 ± 44.00205.95 ± 31.33125.63 ± 29.63129.28 ± 19.95128.45 ± 27.36168.64 ± 44.76
170139.58 ± 36.75220.04 ± 81.29143.37 ± 50.06131.99 ± 35.91145.40 ± 62.42144.06 ± 40.28
1700152.41 ± 20.74196.85 ± 30.44147.68 ± 32.41142.00 ± 43.18144.42 ± 33.26154.43 ± 16.99
3400149.82 ± 31.87219.80 ± 36.34139.27 ± 31.80120.96 ± 21.60122.39 ± 23.45157.52 ± 67.16
Female0199.42 ± 70.03196.41 ± 42.4592.52 ± 22.78108.16 ± 22.97100.79 ± 33.96111.87 ± 37.45
170251.26 ± 119.102 20.78 ± 110.35133.84 ± 46.1499.42 ± 29.84104.54 ± 65.93126.00 ± 56.07
850207.71 ± 65.02192.09 ± 51.63114.62 ± 49.39111.03 ± 34.81101.54 ± 24.52113.47 ± 20.54
1700200.40 ± 66.01154.98 ± 55.67100.97 ± 34.90134.63 ± 61.49118.07 ± 40.20100.22 ± 25.15
Frequency of breathing (bpm)
Male0151.26 ± 28.28181.64 ± 39.36124.50 ± 21.65126.81 ± 13.06125.47 ± 25.24140.68 ± 27.35
170132.55 ± 16.62191.63 ± 52.49136.67 ± 32.41131.53 ± 25.98139.22 ± 37.54127.31 ± 16.17
1700136.98 ± 14.91170.14 ± 30.94134.91 ± 20.48134.08 ± 32.16132.20 ± 27.17133.55 ± 16.26
3400137.29 ± 26.62197.79 ± 40.82138.86 ± 26.02121.07 ± 17.21123.97 ± 21.86133.03 ± 27.51
Female0201.60 ± 71.84198.35 ± 41.89103.29 ± 23.84121.14 ± 24.40114.68 ± 35.22122.23 ± 38.53
170244.77 ± 84.82221.74 ± 50.13139.50 ± 41.87112.48 ± 22.49108.41 ± 35.16138.24 ± 63.74
850202.11 ± 58.91190.68 ± 48.26125.20 ± 48.49121.21 ± 35.33109.03 ± 20.78121.24 ± 20.93
1700202.13 ± 61.13161.13 ± 47.62115.01 ± 31.25143.62 ± 67.68130.99 ± 47.60115.54 ± 24.13
End-Inspiratory Pause (msec)
Male04.29 ± 0.953.58 ± 0.654.31 ± 0.694.18 ± 0.404.19 ± 0.403.83 ± 0.57
1704.33 ± 0.743.61 ± 0.723.84 ± 0.734.27 ± 1.014.34 ± 1.154.06 ± 0.55
17004.36 ± 0.544.43 ± 2.744.93 ± 1.995.08 ± 1.674.50 ± 1.203.95 ± 0.44
34003.98 ± 1.133.21 ± 0.504.36 ± 1.254.44 ± 0.894.29 ± 1.144.36 ± 1.24
Female04.71 ± 1.645.55 ± 3.719.83 ± 7.449.21 ± 6.669.33 ± 6.586.21 ± 1.66
1703.76 ± 1.214.92 ± 3.376.02 ± 1.717.94 ± 3.467.62 ± 3.245.18 ± 2.02
8504.57 ± 1.904.56 ± 1.857.68 ± 3.538.09 ± 3.508.40 ± 4.315.28 ± 1.05
17005.21 ± 2.557.42 ± 6.0410.30 ± 9.159.80 ± 8.629.60 ± 7.426.06 ± 1.96
End-Expiratory Pause (msec)
Male054.13 ± 11.9245.13 ± 10.7861.79 ± 12.8160.39 ± 11.7762.97 ± 13.7855.86 ± 11.24
17057.44 ± 9.5542.32 ± 11.6957.14 ± 14.3959.25 ± 13.8858.90 ± 12.7161.17 ± 8.57
170052.54 ± 8.6044.07 ± 8.0554.77 ± 9.9754.04 ± 9.6455.46 ± 9.4855.81 ± 9.62
340055.48 ± 17.2842.71 ± 13.6856.03 ± 11.7958.43 ± 12.8358.23 ± 12.8759.72 ± 14.38
Female056.52 ± 20.2351.48 ± 11.5778.98 ± 16.3673.63 ± 9.2775.95 ± 13.9178.04 ± 16.29
17049.55 ± 19.3048.79 ± 12.1877.44 ± 28.0682.18 ± 21.7983.97 ± 16.9076.85 ± 33.19
85055.74 ± 15.7247.54 ± 10.4670.23 ± 13.0970.93 ± 10.1475.51 ± 9.1775.08 ± 7.90
170056.87 ± 16.8458.59 ± 15.9870.95 ± 20.0863.04 ± 22.6566.12 ± 15.7976.93 ± 12.53
Enhanced Pause
Male01.73 ± 0.431.43 ± 0.472.00 ± 0.541.91 ± 0.382.10 ± 0.451.72 ± 0.39
1701.89 ± 0.221.35 ± 0.391.89 ± 0.541.94 ± 0.372.08 ± 0.451.98 ± 0.32
17001.57 ± 0.221.28 ± 0.231.65 ± 0.351.72 ± 0.361.70 ± 0.241.67 ± 0.23
34001.79 ± 0.531.24 ± 0.471.85 ± 0.411.91 ± 0.451.96 ± 0.531.87 ± 0.40
Female01.72 ± 0.641.55 ± 0.492.49 ± 0.862.18 ± 0.562.27 ± 0.582.47 ± 0.70
1701.62 ± 0.621.50 ± 0.362.58 ± 0.962.51 ± 0.772.39 ± 0.562.59 ± 1.28
8501.64 ± 0.491.39 ± 0.372.32 ± 0.682.21 ± 0.512.26 ± 0.452.37 ± 0.45
17001.77 ± 0.581.98 ± 0.682.17 ± 0.711.85 ± 0.772.01 ± 0.722.41 ± 0.42
Mid-expiratory flow (mL/s)
Male08.35 ± 1.759.14 ± 0.816.47 ± 1.226.71 ± 1.236.67 ± 1.268.30 ± 2.06
1707.49 ± 1.539.91 ± 3.197.33 ± 1.886.74 ± 1.537.46 ± 2.407.71 ± 2.44
17007.83 ± 1.228.61 ± 1.517.48 ± 1.257.23 ± 1.597.26 ± 1.147.71 ± 1.30
34007.60 ± 1.239.26 ± 1.467.07 ± 1.046.38 ± 0.726.44 ± 0.828.18 ± 3.09
Female09.47 ± 2.688.94 ± 1.995.43 ± 0.635.68 ± 1.035.49 ± 1.356.09 ± 1.30
17012.05 ± 5.7810.29 ± 5.527.12 ± 2.215.73 ± 1.695.89 ± 3.496.66 ± 1.90
85010.36 ± 3.308.48 ± 2.366.25 ± 1.735.90 ± 1.545.61 ± 1.056.27 ± 1.04
170010.06 ± 2.957.34 ± 1.845.54 ± 1.566.90 ± 2.555.99 ± 1.415.38 ± 1.14

Table 3: Effect of T. camphoratus extract on respiratory parameters in rats. All data presented as mean ± SD.

Lin JY and Chiu E. Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Camphoratus Extract. Adv Clin Toxicol 2019, 4(1): 000146.

The beagle dogs were given gelatin capsules containing T. camphoratus extract at doses of 0 (empty capsule), 54, 540, 1000 mg/kg orally and the ECG parameters and blood pressures were measured at indicated time periods. In both sexes, no statistical difference was noted in baseline data of all cardiovascular data. In male dogs, a significant increase in QTcB and QTcF were observed at 4 h at 54 mg/kg, at 3 h and 4 h at 540 mg/kg, at 3 h and 4 h (QTcB only) at 1000 mg/kg as compared to vehicle control. These values, however, were compared to baseline data by paired t-test and no

(Dose)
ParametersGenderPre-dose1 h2 h3 h4 h24 h
mg/kg
RR Interval (ms)
Male0798.413 ± 59.868748.537 ± 28.625764.893 ± 80.637800.077 ± 101.283827.943 ± 115.681716.803 ± 91.396
54578.047 ± 108.284582.813 ± 127.615664.927 ± 163.928617.427 ± 95.058620.623 ± 82.525771.383 ± 130.829
540703.587 ± 186.107683.137 ± 119.233650.053 ± 127.592663.040 ± 119.734601.870 ± 111.554801.173 ± 158.028
1000720.617 ± 169.553569.373 ± 114.235624.447 ± 124.155639.270 ± 69.644598.487 ± 69.776703.330 ± 115.312
Female0706.220 ± 88.543706.773 ± 44.127702.807 ± 114.886703.727 ± 70.382724.317 ± 96.812691.520 ± 123.706
54589.973 ± 102.244549.167 ± 67.720617.430 ± 86.254596.647 ± 66.039642.023 ± 90.679658.123 ± 85.785
540757.533 ± 192.680681.860 ± 154.167739.573 ± 111.680668.697 ± 104.978645.270 ± 108.574757.487 ± 132.135
1000683.033 ± 238.674634.803 ± 174.501648.720 ± 182.701694.740 ± 191.545672.997 ± 189.251661.173 ± 45.326
PR Interval (ms)
Male078.547 ± 7.79377.820 ± 8.33877.073 ± 6.75876.920 ± 7.22883.430 ± 8.87779.360 ± 10.650
5478.453 ± 5.90578.503 ± 6.07979.927 ± 6.03079.787 ± 6.90083.517 ± 9.69280.597 ± 5.993
54081.067 ± 5.26184.453 ± 8.48679.307 ± 7.15182.943 ± 7.96481.617 ± 9.11579.240 ± 6.221
100081.887 ± 7.76882.583 ± 8.16682.740 ± 6.85980.860 ± 4.77979.970 ± 4.49385.890 ± 7.518

Table 4: Effect of _T. camphoratus_ Extract on ECG Parameters in Beagle Dogs. All data presented as mean ± SD. *_p_< 0.05 [Statis

Lin JY and Chiu E. Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Camphoratus Extract. Adv Clin Toxicol 2019, 4(1): 000146.

Female093.587 ± 6.35185.920 ± 2.88288.937 ± 8.37187.247 ± 5.86487.880 ± 8.31589.810 ± 3.775
5491.630 ± 6.39589.163 ± 3.99588.340 ± 2.40985.183 ± 3.44591.230 ± 7.96291.010 ± 5.254
54090.647 ± 7.09789.200 ± 3.81090.050 ± 8.08785.137 ± 0.70188.273 ± 4.07790.087 ± 4.650
100099.893 ± 12.18495.203 ± 8.70994.670 ± 11.69092.050 ± 6.23190.963 ± 8.57092.100 ± 4.723
P wave Duration (ms)
Male035.770 ± 5.35034.847 ± 4.06435.080 ± 4.05236.190 ± 3.42842.330 ± 4.47635.680 ± 7.207
5435.087 ± 3.26435.843 ± 3.36037.133 ± 3.92837.340 ± 4.68840.470 ± 3.78536.537 ± 4.505
54039.120 ± 4.79343.760 ± 3.05037.997 ± 5.21344.170 ± 5.11640.577 ± 3.13137.117 ± 4.885
100036.080 ± 4.40237.693 ± 3.59237.277 ± 3.40637.680 ± 3.96436.847 ± 4.05238.497 ± 3.124
Female037.870 ± 2.10236.037 ± 1.40937.877 ± 2.41036.637 ± 2.24936.790 ± 2.10238.213 ± 2.027
5437.043 ± 6.30035.997 ± 2.88435.347 ± 4.11934.120 ± 3.10838.560 ± 8.98236.377 ± 3.851
54038.673 ± 2.79136.793 ± 2.01038.900 ± 4.94835.290 ± 2.26635.320 ± 4.15735.713 ± 3.203
100041.470 ± 4.93639.493 ± 4.00340.280 ± 5.97439.307 ± 4.15737.907 ± 5.97237.160 ± 2.833
QRS wave Duration (ms)
Male040.737 ± 3.34039.530 ± 1.88239.440 ± 1.38439.010 ± 2.65338.933 ± 3.62239.983 ± 2.174
5439.333 ± 1.44439.197 ± 2.11339.933 ± 1.82639.103 ± 1.98540.913 ± 4.18039.880 ± 1.228
54039.610 ± 3.87938.920 ± 3.91639.927 ± 3.43539.080 ± 3.98139.963 ± 3.59140.413 ± 4.524
100038.860 ± 2.22339.050 ± 1.25439.387 ± 1.03039.650 ± 2.00139.280 ± 1.50839.867 ± 2.140
Female040.853 ± 1.95538.577 ± 2.91639.417 ± 4.52538.740 ± 3.24838.620 ± 2.02542.017 ± 6.946
5440.373 ± 4.67437.230 ± 2.08938.600 ± 3.21240.270 ± 5.96337.597 ± 2.75238.023 ± 3.410
54040.600 ± 3.18238.343 ± 2.36139.367 ± 0.73138.863 ± 2.48639.053 ± 2.75439.670 ± 1.573
100037.693 ± 2.54637.073 ± 1.84837.720 ± 1.34037.073 ± 1.40936.610 ± 2.20137.757 ± 2.214
QT Interval (ms)
Male0206.057 ± 1.141201.877 ± 1.916206.367 ± 5.905203.893 ± 2.259202.930 ± 3.292210.413 ± 6.634
54191.583 ± 14.820186.527 ± 11.450196.833 ± 12.281195.250 ± 9.689201.210 ± 9.257202.630 ± 5.552
540204.273 ± 20.341202.880 ± 7.816200.253 ± 12.000206.530 ± 9.097197.860 ± 10.753205.930 ± 9.756
1000201.030 ± 12.067188.980 ± 10.458195.600 ± 8.354202.487 ± 9.031196.073 ± 4.124210.107 ± 4.766
Female0204.360 ± 9.544190.747 ± 6.205199.027 ± 18.670197.817 ± 10.616198.280 ± 12.799203.600 ± 19.481
54195.887 ± 12.476183.640 ± 12.736191.120 ± 8.444187.437 ± 6.908198.163 ± 12.725199.873 ± 11.167
540213.013 ± 27.911198.193 ± 20.540207.780 ± 31.052196.570 ± 8.557199.530 ± 16.356209.890 ± 20.110
1000202.463 ± 30.136197.110 ± 26.206195.013 ± 19.255201.963 ± 29.364200.973 ± 30.712200.737 ± 20.575
QTcB (ms)
Male0242.907 ± 6.692243.660 ± 6.197248.547 ± 5.228241.397 ± 9.210236.070 ± 9.328258.943 ± 15.129
54258.383 ± 8.989251.897 ± 13.068252.303 ± 12.607256.753 ± 5.337263.070 ± 7.214 #242.643 ± 6.930
540254.887 ± 1.825255.627 ± 8.920257.947 ± 7.875263.823 ± 10.519 #263.713 ± 6.470 #240.467 ± 10.800
1000250.220 ± 8.885256.427 ± 12.486255.067 ± 15.687260.670 ± 4.626 #260.097 ± 8.056 #259.977 ± 16.944
Female0256.907 ± 19.104236.567 ± 10.190249.990 ± 20.813247.377 ± 15.665245.403 ± 14.907256.907 ± 4.757
54261.820 ± 14.408254.100 ± 10.237252.130 ± 9.935249.173 ± 10.426257.200 ± 20.161256.577 ± 12.554
540259.277 ± 32.079250.763 ± 19.598253.230 ± 27.836251.100 ± 16.082258.127 ± 8.173252.877 ± 9.634
1000258.943 ± 9.687257.490 ± 5.664252.857 ± 8.804254.310 ± 16.623255.803 ± 14.263253.970 ± 17.136
QTcF (ms)
Male0228.973 ± 4.524228.053 ± 4.147232.630 ± 4.046227.137 ± 5.765223.437 ± 5.206240.823 ± 9.602
54233.347 ± 7.490227.303 ± 6.329231.440 ± 5.177233.730 ± 1.535239.993 ± 3.820 #227.633 ± 3.101
540235.830 ± 6.452235.900 ± 2.805236.320 ± 2.015242.367 ± 4.452 #238.987 ± 0.655 #227.510 ± 3.836
1000231.650 ± 4.715231.150 ± 4.893232.903 ± 7.997239.080 ± 4.492 *236.260 ± 3.948 #241.460 ± 9.898
Female0237.030 ± 14.782219.433 ± 8.118230.733 ± 18.505228.717 ± 13.100227.553 ± 12.429236.720 ± 10.037
54237.153 ± 10.328227.547 ± 10.176229.223 ± 6.562226.117 ± 6.983235.077 ± 16.229235.297 ± 10.258
540241.640 ± 26.874230.963 ± 16.278236.070 ± 28.170230.620 ± 11.319236.103 ± 9.233236.650 ± 11.816
1000237.367 ± 12.446234.683 ± 12.643231.050 ± 6.433234.493 ± 18.764235.080 ± 18.502234.250 ± 18.580
Heart Rate (bpm)
Male075.430 ± 5.61580.237 ± 3.13479.000 ± 7.96775.750 ± 8.96373.417 ± 10.21384.697 ± 11.660
54106.190 ± 19.256106.157 ± 22.12393.740 ± 21.48398.720 ± 15.09297.927 ± 14.09979.180 ± 12.379
54088.873 ± 20.39089.490 ± 14.26094.827 ± 19.41392.583 ± 17.472102.090 ± 19.53376.803 ± 14.618
100086.330 ± 19.731108.670 ± 24.61398.937 ± 21.58794.643 ± 10.827101.200 ± 12.23286.973 ± 15.244
Female085.930 ± 11.60885.123 ± 5.49687.067 ± 15.55585.860 ± 9.01883.920 ± 12.15188.493 ± 14.495
54103.667 ± 17.091110.410 ± 14.06998.553 ± 14.795101.367 ± 10.89294.830 ± 14.58492.300 ± 12.997
54083.207 ± 23.73891.260 ± 21.82582.417 ± 12.84491.350 ± 15.52494.813 ± 16.37280.723 ± 13.057
100096.020 ± 35.79798.977 ± 24.47997.157 ± 25.05690.943 ± 25.24793.757 ± 24.83091.040 ± 6.416

Table 5: Effect of _T. camphoratus_ Extract on ECG Parameters in Beagle Dogs. All data presented as mean ± SD. *_p_< 0.05 [Statis

Lin JY and Chiu E. Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Camphoratus Extract. Adv Clin Toxicol 2019, 4(1): 000146.

Table 4: Effect of T. camphoratus Extract on ECG Parameters in Beagle Dogs. All data presented as mean ± SD. *p< 0.05 [Statistical difference from the control group by Dunnett’s method and from baseline by paired t-test.] #p< 0.05 [Statistical difference from the control group by Dunnett’s method, but with no statistical difference from the baseline by paired t-test.].

(Dose)
ParametersGenderPre-dose1 h2 h3 h4 h24 h
mg/kg
Diastolic Arterial Pressure (mmHg)
Male071.723 ± 10.71774.730 ± 5.69772.787 ± 17.09585.827 ± 22.94371.780 ± 9.82072.033 ± 8.915
5475.653 ± 13.46284.387 ± 31.29264.973 ± 13.56972.243 ± 8.95472.173 ± 16.36873.593 ± 6.770
54070.243 ± 6.93467.113 ± 5.57765.300 ± 10.88867.040 ± 2.74675.883 ± 10.16772.167 ± 4.491
100064.037 ± 8.87882.787 ± 33.93272.613 ± 27.03762.340 ± 13.23254.913 ± 8.35864.083 ± 7.796
Female067.503 ± 12.80482.767 ± 10.91974.023 ± 4.97268.727 ± 5.41667.130 ± 9.91870.747 ± 14.513
5461.410 ± 5.57770.997 ± 9.35261.027 ± 9.09568.193 ± 5.44381.777 ± 26.26364.497 ± 6.579
54064.857 ± 4.94566.803 ± 1.58965.270 ± 7.665 a76.647 ± 15.12661.960 ± 8.82563.763 ± 4.128
100064.527 ± 8.05771.050 ± 8.44065.943 ± 6.65956.537 ± 10.00960.953 ± 7.54359.430 ± 15.599 a
Systolic Arterial Pressure (mmHg)
Male0119.173 ± 19.173129.633 ± 15.506123.827 ± 28.079132.487 ± 21.398116.603 ± 7.987119.427 ± 18.234
54122.290 ± 15.031140.467 ± 30.572126.777 ± 21.013123.560 ± 10.1391 23.687 ± 15.826124.293 ± 11.099
540126.227 ± 16.320121.060 ± 5.983120.343 ± 17.277121.477 ± 15.6231 25.267 ± 12.978127.173 ± 19.665
1000108.950 ± 6.190123.440 ± 24.995118.163 ± 18.314104.717 ± 7.843109.430 ± 10.546115.063 ± 16.114
Female0106.443 ± 10.599124.920 ± 11.499121.900 ± 16.123116.410 ± 13.1261 20.020 ± 22.471116.490 ± 10.157
54105.440 ± 5.856116.863 ± 12.171105.197 ± 6.617110.187 ± 4.226128.307 ± 32.081109.897 ± 11.591
540104.160 ± 18.689115.327 ± 9.046111.595 ± 9.595 a115.917 ± 11.057102.357 ± 5.889101.993 ± 9.488
1000103.693 ± 5.809111.997 ± 11.882110.540 ± 12.109103.937 ± 8.532103.387 ± 6.739106.865 ± 9.991 a
Mean Arterial Pressure (mmHg)
Male093.233 ± 14.94399.557 ± 11.07997.340 ± 25.772106.733 ± 23.82391.627 ± 9.78293.693 ± 14.800
5496.060 ± 11.596111.737 ± 31.78790.910 ± 9.94593.963 ± 10.44094.610 ± 16.97397.237 ± 7.714
54097.303 ± 11.18494.377 ± 6.96388.163 ± 14.13193.830 ± 12.64798.510 ± 11.62199.267 ± 17.554
100086.113 ± 5.078100.857 ± 30.11892.763 ± 23.17781.473 ± 10.42673.910 ± 8.95884.627 ± 10.630
Female084.833 ± 12.458100.740 ± 9.97295.927 ± 9.89989.720 ± 7.92288.547 ± 13.67490.990 ± 10.507
5481.087 ± 6.15391.720 ± 10.41481.060 ± 7.45388.200 ± 5.587101.930 ± 27.70385.550 ± 9.611
54082.320 ± 10.84988.760 ± 3.63187.025 ± 8.224 a95.253 ± 14.12579.360 ± 5.32380.507 ± 8.033
100082.310 ± 5.91688.903 ± 10.70785.977 ± 9.41477.630 ± 9.76880.873 ± 6.40376.955 ± 17.458 a

Table 6: Effect of _T. camphoratus_ Extract on Blood Pressure in Beagle Dogs. All data presented as mean ± SD. a : N = 2 (The pea

Table 5: Effect of T. camphoratus Extract on Blood Pressure in Beagle Dogs. All data presented as mean ± SD. a : N = 2 (The peak blood pressure of one female at 540 mg/kg was unrecognizable during 1 to 2 h post-dosing period and cannot be calculated reliably by iox system. In addition, no blood pressure was recorded in one female at 1000 mg/kg during 23 to 24 h post-dosing period, because the tail cuff was loose. Therefore, the data were not included in statistical calculation.) The ECG morphology at each indicated time period was evaluated by a veterinary cardiologist that the sinus arrest noted in all six dogs and large negative T wave Lin JY and Chiu E. Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Camphoratus Extract. Adv Clin Toxicol 2019, 4(1): 000146.

noted in two dogs before and after the T. camphoratus extract treatment (data not shown). In general, T wave changes are very non-specific. Tall T wave could be as a normal variant. This could occur with hyperventilation, anxiety, and even positional changes. However, tall T wave could also be a warming sign of myocardial hypoxia or electrolytes disturbance (hyperkalemia). Sinus arrest is frequently caused by high parasympathetic tone due to one or many factors. It is commonly in brachycephalic breed dogs with strenuous respiratory efforts that irritate the pharynx and cause reflex vagal stimulation. Other factors may cause sinus arrest include surgical stimulation, impingement upon the vagus nerve (neoplasia), drug toxicity (digitalis or β-blockers). No treatment is needed for this conducting disturbance, unless syncope is developed [18, 19]. In this study, large (negative) T wave was found in one male and one female dog and sinus arrest was seen in all six dogs in both the predose and post stages. The link between these abnormal findings and T. camphoratus extract was not indicated. Based on the results, the dogs received T. camphoratus extract via oral administration up to the dosage of 1000 mg/kg did not cause physiological abnormalities on cardiovascular system in this study.

Conclusion

Results from in vitro hERG test and in vivo core batteries of safety pharmacology studies revealed that T. camphoratus extract had no significant effect on the hERG current at nominal concentrations of up to 25 μg/mL and had no obvious toxicity evidences on central nervous, respiratory and cardiovascular system (up to doses at 1700 mg/kg in female rats, 3400 mg/kg in male rats and 1000 mg/kg in both sexes of dogs). The results would provide the evidences to support the safety of T. camphoratus extract as a food supplement and for clinical usage.

Conflict of Interests

The authors declare that there are no conflicts of interests.

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@article{lin2019,
  title   = {Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Extract},
  author  = {Lin JY and Chiu E},
  journal = {Advances in Clinical Toxicology},
  year    = {2019},
  volume  = {4},
  number  = {1},
  doi     = {10.23880/act-16000146}
}
Lin JY and Chiu E (2019). Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Extract. Advances in Clinical Toxicology, 4(1). https://doi.org/10.23880/act-16000146
TY  - JOUR
TI  - Preclinical Safety Pharmacology Studies of Taiwanofungus camphoratus Extract
AU  - Lin JY and Chiu E
JO  - Advances in Clinical Toxicology
PY  - 2019
VL  - 4
IS  - 1
DO  - 10.23880/act-16000146
ER  -