Journal of Medicinal Materials, 2015, VoL 20, No. 5 (pp.291-296)
SAFETY AND TOLERABILITY OF AMA KONG CAPSULES:
A PHASE I TRIAL
Huynh Thi Thuy Trang', Bui Thi Huong Quynh', Nguyen Thi Bay', Nguyen Minh Duc''^ * University of Medicine and Pharmacy, Ho Chi Minh City, Vietnam
^Ton Due Thang University, Ho Chi Minh City, Vietnam
*Corresponding author: [email protected] (Received May, 3"*, 2015)
Summary
Safety and Tolerabiiity of Ama Kong Capsules:
A Phase I Trial
The aims of this study were to evaluate the safety and tolerabiiity of Ama Kong capsules in healthy volunteers. We conducted an open, non-placebo controlled trial, monitored before and after using Ama Kong capsules containing 150 mg of Ama Kong dry extract powder par capsule. Three doses including 2, 4 and 6 capsules/day within 14 days were used to assess safety and tolerabiiity. The study subjects included 29 healthy male volunteers aged 20-30 years, were randomized into 3 groups. Routine laboratory tests and physical examinations were performed before and after the first single dose and weekly during a 14-day follow-up period to observe potential adverse effects. The results showed no significant changes in laboratory data for hematology values, urine analysis and prostate ultrasound examination in all 3 groups. Testosterone levels in group using 4 capsules/day increased significantly after a 14-day period of trial (p<0.05). Hence, the phase I trial demonstrated that Ama Kong capsules with doses of 2 capsules/day, 4 capsules/day and 6 capsules/day were safe and well- tolerable in healthy male volunteers.
Keywords: Ama Kong capsules. Dose, Tolerabiiity, Safety.
1. Introduction
Male sexual dysfunction is a common sexual problem in men, and becomes more common with increasing age. It can be caused by psychological or physical problems, such as lowered levels of male testosterone. The American Association of Clinical Endocrinologists (AACC) recommended testosterone replacement treatment, phosphodiesterase type 5 inhibitors PDE5 (sildenafil, tadalafil, vardenafil), and prostaglandin El in current guidelines for medication treatment of MSD [1]. However, these drugs can induce multiple serious adverse effects, and thus new drugs that have good tolerabiiity and safety are required to treat MSD. Recently, many natural products have been used to increase testosterone levels in men who suffer from MDS [2, 6]. In this regard, Ama Kong's remedy is one of the most common traditional ones. Ama Kong capsules are manufactured based on the extract of three Vietnamese medical plants, Tam Ngleng (L) in
the Family Rutaceace, Nam Dong (D) in the Family Smilacaceace, and Tom Trong Nenso (T) in the Apocynmem. Several previous phamiacological studies showed that Ama Kong's remedy exerted nutritious and tonic effects, and increased male hormone levels [4, 5, 6].
The aims of this study were to evaluate the safety and tolerabiiity of three oral doses of Ama Kong capsules, 2 capsules/day; 4 capsules/day, and 6 capsules/day in healthy volunteers.
2. Materials and methods
Ama Kong capsules manufactured by Pharmaceutical company 3/2, each capsule contains
150 mg of Ama Kong dry extract powder, labeling "Drug for clinical trial. Do not be used for another purpose" were used in this study
Healthy male volunteers, aged 20-30 years with body mass index of 19-23 kg/m^ were eligible for the trial. Criteria included: no clinically abnormal findings on history particularly liver and kidney diseases, all values
Journal of Medicinal Materials, 2015, VoL 20, No, 5
of hematological and biochemical tests within normal ranges, no use of any other drug in the past 30 days. People who had a history of alcoholism or non-compliance were excluded from the study. All volunteers were informed of the nature of the study and provided written informed consent prior to participation [3, 7].
The Ethics Committee of the University Medical Hospital at Ho Chi Minh city approved the study.
At enrolment, a medical history was taken, including a full physical examination, prostate ultrasound, and routine laboratory investigations (biochemistry, complete blood count, urinalysis).
Volunteers were randomly allocated to one of the three groups. Volunteers from the first group
received 2 capsules/day. The second group received 4 capsules/day (2 capsules twice a day).
Volunteers fi^m third group received 6 capsules/
day (2 capsules three times a day) for 14 days.
The study drugs were administered after meals.
Three groups were given drug subsequentially.
Each dose and timing of administration was documented. Study volunteers were visited 8 times during a 14-day follow-up period of taking drug and adverse effects of drug were recorded.
The trial will be immediately discontinued if volunteer has allergy or sensitivity to the study drug or inability to tolerate the study drug. Table I and 2 present scheduled clinic visits and assessments.
Table 1. Scheduled clinic visits Visit number
Visit 1 Visit 2 Visit 3 Visit 4 Visit 5 Visit 6 Visit 7 Visit 8
Type of visit Information, Screening & Consent (day 0)
Randomization in 3 groups. Single dose was given. Follow-up after 1 Laboratory test 24 hours after the single dose (day 1) Start the 14-day course of taking dmg Follow up after 3 days of drug use
and 2 hours
Follow up aft« 7 days of drug use (day 7) Follow up after 10 days of drug use Follow up after 14 days of drug use (day 14)
Table 2. Visits and assessments Visit number
Informed consent Medical/ smol(ing tiistoiy Inclusion/Exclusion criteria Physical examination, Heigtit and weigtit Vital signs: heait rate, blood pressure, etc Laboiatoty tests: Hematology & biochemistry:
Urinalysis Testosterone Adverse events
Study medication - Dispensed/Returned (D/R) Diary - Dispensed/Returned
1 X X X X X X X X X
2
X X
X D
3
X X X X X
4
D D
5
X X
X R/D R/D
6
X X X X X R/D R/D
7
X X
X R/D R/D
8
X X X X X X R R Seven ml of blood samples and 10 ml of urine
samples were collected at screening (visit I), on day 1 after single dose, day 7, and 14 of 14-day course of drug use. Following tests were performed:
Serum biochemistry profile (blood urea nitrogen, creatinine, aspartate transaminase (AST), alanine transaminase (ALT), blood glucose); hematology profile (red blood cell count, hemoglobin, white
blood cell count with differential, and platelet count); urine analysis (urobilinogen, spe-gravity, pH); prostate ultrasound. Testosterone levels were checked on the screening day and after the last day of drug use.
The volunteers were physically examined and adverse reactions during the study were recorded with the date and time at which they occurred and
292 Journal of Medicinal Materials, 2015, VoL 20, No. S
disappeared. Adverse effects may be manifested as disturbances in multiple body organs: skin (allergy, skin rash), gastrointestinal tract (nausea, vomiting, diarrhea); central nervous system (headache, dizziness, insomnia); cardiovascular system (palpitations, chest pain), arthralgia, nocturia and others.
Statistical analysis was performed using MS Excel and SPSS 16.0. Data were presented as mean ± SD (standard deviation). We used Kruskal-Wallis test to compare differences in each item of the laboratory data at each follow- up, with respect to data baseline levels more than two independent groups by. If this showed the difference, differences between two independent groups continue to be compared in pairs by Mann Whitney-test, p < 0.05 was considered statistically significant difference.
3. Results
At enrolment 29 volunteers were included and followed up in the study, randomly assigned to three p^ups - 11 voluteers in group 1, 10 in group 2, and 8 in group 3. All study volunteers were healthy, as verified by laboratory results, vital sign, monitoring, physical exam, prostate ultrasound and testosterone test. No significant differences in the distribution of demographic, and most of the clinical and laboratory data between the three groups, excepted data of WBC (p = 0.019), AST (p = 0,015), ALT (p = 0,015).
However, all of the baseline laboratoiy data and baseline vital signs were within normal ranges.
Laboratory data for tiie 3 groups of healtiiy volunteers during the sftidy WCTB shown in Tables 3,4,5.
Groi^ I. No saious adverse events were (Asaved, nine of 11 volunteers had no adverse effects during drug use. Some volunteers complained of uncomfortable symptoms during drug administration, as follows; mild headache (1 case), abdominal pain (1). Two symtoms were mild and disappeared spontaneously. The results showed no significant changes in laboratoiy data for hematology values and urinalysis (both within-group, and between- group effects). All prostate ultrasound results were normal. Testosterone levels after 14 days
increased compared with those before drug use (662.0 ± 152.8 vs 586.3 ± 157), but had no significant difference (p= 0.3). Levels of glucose, AST and ALT changed significantly but were still in the normal ranges. In summary, a dose of 2 capsules/day showed safety and tolerabiiity in healthy volunteers. We continued to study in the group 2.
Group 2. No serious adverse events were observed, six of 10 volunteers had no adverse effects during drug use. Some volunteers complained of unpleasant symptoms during drug administration, as follows; mild restless (1 case), mild abdominal pain (2), insomnia (1). All symtoms were mild and disappeared spontaneously.
The results showed no significant changes in laboratory data for hematology and biochemistry values as well as urinalysis (both within-group, and between-group effects). All prostate ultrasound results were normal. The testosterone levels after 14 days increased significantly (p=0.023), compared with those before drug use (793.5 ± 196.7 vs 637.3 + 148). Our data suggest that, a dose of 4 capsules/day are safe and tolerable in healthy volunteers. We continued to study in the group 3.
Group 3. No serious adverse events were observed, five of eight volunteers had no adverse effects during drug use. Some volunteers complained of abnormal symptoms during drug administration, including palpitation (2 cases) and abdominal pain (1). All symtoms were mild and disappeared spontaneously. The results showed no significant changes in laboratoiy data for hematology values and urinalysis (both within-group, and between-group effects). All prostate ultrasound results were normal. The levels of BUN and AST changed significantly but still in the normal ranges. Testosterone levels after 14 days increased compared with those before drug use (664.4 ± 197.0 vs 644.9 ± 102), but the difference was not statistically significantly (p = 0.88). These findings documented the safety and tolerabiiity of the study dmg with a dose of 6 capsules/day in healthy volunteers.
Journal of Medicinal Materials, 2015, VoL 20, No. 5
Table 3. Laboratory data of 11 volunteers (group 1) durmg study
Table 4. Laboratory data of 10 volunteers (group 2) during study BMI (kg/nr)
Systolic pressure (mmHg) Diastolic pressure (mmHg) Pulse (beats per minute) Temperature ("C)
Respiratory rate (breaths per minute]
Hematology WBC(xlO'/L) RBC(xlO"/L) MCV (ft,) MCH (pg) MCHC (g/dl) Biochemistry Glucose (mmol/l) BUN(mg/100m!) GFR (ml/min) AST (u/l) ALT (u/l) Urinalysis Urobilinogen (E.U./dl) Spc-Gravity (cells/uL) pH
Testosterone (ng/dl)
Day 0 (At enrolment) 20.2 ±2.3 I12.9±8.l 7I.8±6.0 71 8 ±7.8 36.3 ±0.2 I9.6±1.5 7.1 ±1.4 5.1 ± 0 2 86.7 + 2 8 28.2 ± 0 7 32.6 ± 0.6 245.3 ± 33.9
4.6 ± 0.3 32.3 ± 7.4 136.2 ±22.4 19.0 ±4.2 I4.0±2.l 0.25 ±0.1 1.02 ± 0 0 5.9 ± 0 5 586.3 ±157
Day 1 20.2 ± 2 5 117.6 ±11.9
70.6 ± 7.7 70.6 ± 6.3 36 2 ±0.1 18.9 ± 1.8 6.8 ±0.2 4.9 ± 0.2 86 8 ± 2.6 28.3 ±0.6 32.6 ±0.5 237.9 ±35.2
4.1 ±0.2 27.9 ±6.5 126.6 ±36.1
27.0±5 3 24.9 ±7.8 0.2 ± 0.0 1.02 ±0.0 6.I8±0.0
Day?
20.2 ±2.1 118.5 ±12.6
68.1 ±9.9 73.6 ± 10 4 36.1 ±0.2 19.0 ±1.3 7.0 ± 1 3 4.9 ± 0.2 86.9 ± 2 4 28.4 ± 0 5 32.7 ±0.4 241.0 ±29.5
4.8 ± 0 2 25 2 ± 8.0 127.8 ±25.3 26 6 ± 5.7 22.9 ±7.1 0.2 ±0.0 1.01 ±0.0 6 1 8 ± 0 0
Day 14 20.2 ±2.3 120.7 ±12.4 72.8 ± lO.I 76.0 ±10 0 36.4 ± 0.3 18.5 ±1.5
7.5±2.1 5.0 ± 0.2 86 7 ±0.8 28 4 ±0.5 32 8 ±0.3 248.0 ±27.8
4.5 ± 0.3 29.5± 3.8 I10.3±I3.6
22.1+4.6 17.4 ±4.5 0.2 ±0.0 1.01 ±0.0
6.0 ±0.0 662.0 ± 152.8
Rvalue 0.920 0.430 0.624 0.52S O.ISI 0.344 0.850 0.221 0.989 0.908 0.796
0.029 0.134 0.147 O.OOS 0.000
10 0.190 0.307 0.300
BMI (kg/m') Systolic pressure (mmHg) Diastolic pressure (mmHg) Pulse (beats per minute) Temperature ("C)
Respiratory rate (breaths per minute) Hematology
WDC(xlO'/L) RBC(xlO'-/L) MCV (IL) MCH(pg) MCHC (g/dl) PLT(xlO''/L) Biochemistry Glucose (mmol/1) BUN(mg;iOOml) GFR (ml/min) AST (u/l) ALT (u/l) Urinalysis
Day 0* (before drug use) 20 5 ± 2.0 119.1 ±6.0 66.4 ±3.8 36.3 ± 0.4 I8 2±1.7 5.5 ±1.2 5.1 ±0.2 86 7 ± 2.8 28.4 ±1.0 32.3 ±0.5 223 1 ±38.4
5.0 ± 0.4 31.6±5.8 126.2 ±25 7
26.9 ±8.1 2 4 7 ± l l . l
Day 1 205 ± 2 1 I14.0± 11.6 64.4± 6.3 72.4± 5 3 36.3 ±0.3 18.2 ±1.4 5.9 ±1.4 4.9 ± 0.2 86.8 ± 2.6 26.3 ± 0.7 32.3 ±1.5 226.1 ±38.6
4.8 ±0.2 27.7 ±5.2 134.2 + 22.5
26 3 ± 6.4 26.1 ± 1 1 5
Day 7 20 5 ± 2.4 115.1 ± 6 9 64.3± 7 i 71.2±8.l 36.3 ± 0 2
I8.2± 1.4 5.7 ±1.0 4.9 ± 0 2 86.9 ±2.4 28.3 ± 0.4 3 2 2 ± l . 4 216.6 ±46.5
4.5+0.4 30 1 ± 5 6 132.6 ± 23.4 22.3 ±4.7 I 9 . 7 ± 7 6
Day 14 20 5 ± 2.3 113.1 ±7.5 62 5 ± 6.2 74.2 ±9.0 36.4 ± 0.3
17.8±l.l 5.5±l.l 5.0 ±0.2 86.7 ± 0 8 28.7 ±0.7 32.5 ± 0 8 223.8 ± 50.5
4 8 ± 02 34.2 ±4.9 119 9 ±14.8
19.8 ±8.1 20.9 ± 10.7
P value 0.945 0.375 0.582 0.101 0.747 0.887 0.934 0.061 0.953 0.750 0.903 0.864 0.052 0.087 0.459 O068 0 493
294 Journal of Medicinal Materials, 2015, VoL 20, No. 5
Urobilinogen (E.U./dl) SpcOiavity (cells/uL) pH
0.2 ±0.0 1.02 ±0.0 6.2 ±0.3
0.2 ±0.0 1.04 ±0.0 6.03 ±0.1
0.2 ±0.0 1.02 ±0.0 6.21 ±0.3
0.2 ±0.0 1.01 ±0.0 6.10±0.3
1.000 0.174 0.493 Testosterone (ng/dl)
Table 5. Laboiatory data of 8 voluntecis (group 3) during study
BMI (kg/m^) Systolic pressure (mmHg) Diastolic pressure (mmHg) Pulse (beats per minute) Temperature (°C)
Respiratory rate (breaths per minute) Hematology
WBC (xlO'/L) R B C ( X I O ' V L ) MCV (IL) MCH (pg) MCHC (g/dl) PLT(xl0'/L) Biochemistry Glucose (mmol/1) BUN (mg/lOOmI) GFR (ml/min) AST (u/l) ALT (u/l) Urinalysis Urobilinogen (E.U./dl) Spe-Gravity (eells/uL) pH
Testosterone (ng/dl)
Day 0* (before dmg use) 21.3 ±2.3 123.7 ± 9.2 68.5 ± 7.8 74.6 ±10.6 36.3 ± 0.3 19.0±1.0 5.3 ±1.0 4.8 ± 0.3 88.7 ±1.9 28.9 ±0.6 32.6 ±0.4 219.5± 43.7
4.5 ±0.4 24.8 ±4.6 130.3 ± 35.4
26.2 ± 4.5 25.8 ±8.9 0.2 ±0.0 1.02 ± 0.0 5.9 ±0.6 644.9 ±102
Day 1 21.3 ± 2 5 116.6 ±10.2 66.2 ±6.4 70.0 ±9.9 36.5 ± 0.2 18.5 ±(0.9) 5.6 ±0.6 4.6 ±0.3 89.3 ±1.8 28.8 ±0.3 32.3 ±4.1 211.3±37.9
4.2 ±0.2 30.9 ± 5.9 I10.l±28.4 24.3 ± 5.2 18.6 ±4.5 0.2 ±0.0 1.0I±0.0 6.18±0.5
Day?
21.3 ±2.6 112.5 ±7.2 67.0 ±6.7 69.0 ±8.0 36.5 ±0.1 18.2 ±0.7 5.6 ±0.8 4.6 ±0.3 89.0 ±3.2 29.1 ±0.2 32.7 ± 3.4 223.3 ± 36.8
4.6 ±0.4 33.8 ± 5.2 117.3 ± 25.6
20.9 ±4.5 17.1 ±5.8 0.2 ±0.0 1.01 ±0.0 6.31 ±0.7
Day 14 21.4±2 7 1I8.3±9.1 68.6 ±7.0 73.2 ± 10.8
36.4 ±0.2 19.0±I.O 5.5 ±1.0 4.7 ±0.3 88.4 ± 2.4 29.0 ±0.4 32.7 ±4.2 223.2 ± 40.1
4.7 ±0.2 30.0 ±5.5 128.2 ±20.8
16.8 ±5.8 18.1 ±8.3 0.2 ±0.0 I.0I±0.0 6.4 ±0.6 664.4 ± 197.0
P value 0.897 0.127 0.951 0.603 0.446 0.305 0.689 0.677 0.351 0.836 0.412 0.802 0.059 0.028 0.346 0.011 0.144 I.OOO 0.676 0.538 0.878 4. Discussion
The present phase I trial demonstrated that the usage of Ama Kong with doses of 2-, 4-, and 6 capsules/day during a 14-day period was safe and tolerable among 29 healthy volunteers. Some volunteeis complained about uncomfortable ^mptoms during drug administration in the study, but all of the symtoms were mild and disappeared spontaneously. Nausea and vomiting were not observed during study. All of the symptoms resolved at the end of drug administration. It is difficult to distinguish abnormal symptoms that were not related to the study drug and study drug- related side-effects. Laboratoiy monitoring showed variations each follow-up day, but all data were within normal ranges.
The testosterone levels in volunteers of 3 groups tended to increased, but only in group 2, the change in this level was significant (p = 0.023).
The mechanism of testosterone change after using Ama Kong capsules was not investigated, so it must be controlled carefully during drug use, especially in the people with high baseline testosterone level.
5. Conclusion
The phase I trial demonstrated the safe^ and tolerabili^ of Ama Kong capsules in three doses of 2 capsules/day, 4 capsules/day and 6 capsules.
A dose of 4 capsules/day may increase testosterone level. Further investigation should be done to clarify the pharmacokinetic profiles of Ama Kong capsules.
1. American Association of Clinical Endocrinologists. Medical guidelines for clinical practice for the evaluation ud treatment of male sexual dysfunction: a couple's problem-2003 update. 2. Ang H. H., Lee K. L.. Kiyoshi M. (2004), Sexud arousal in sexually slug^sh old male rats after oral admmistration of Eurycoma longifolia Sack, Journal of Basic and Omical Physiology and Pharmacology, 15(3-4X 303-9. 3. Nell D. H. (2005), A Safety study of sutheriandia frutescens, a South African medicinal plant, in healthy adult volunteers. Clinical Trials, 53(7), 235-241. 4. Nguyen Minh Due. Nguyen Ngoc Khoi, Le Thi Thuy Hang. Do Van dung. Ho Viet Sang (2008), Ama Kong's Remedy, A folk Vietnamese herbal fonriula.
increases endurance swimming capacity of Mice, Journal of Medicinal Materials, 13(6), 292-296. 5. Nguyen Ngoc Khoi, Nguyen Viet Ha. Nguyen Minh Due (2009), Effects of Ama Kong's remedy on growth of accessory sexual organs and plasma testosterone levels in Mice. Journal of Medicinal Materials, 14(1), 52-56. 6. Nguyen Thi Thu Huong, Tran My Tien, Tran Cong Luan, Nguyen Minh Due (2009), Androgenic Effects of Ama Kong's remedy on castrated mice. Journal of Medicinal Materials, 14(2), 108-114. 7. Noppadon yangpukdee. aungkana hiransaree, srivicha krudsood, kobsiri chalennra, siripun srivilairit, wattana leowattana, somsong rakpao, somchai looareesuwan (2004), Phase 1 clinical trial to evaluate the safety and tolerabiiity of oral SN-1 extract formulation in healthy thai volunteers, Journal of Tropical Medicine andParasitology.27(2). 71-8.
Journal of Medicinal Materials, 2015, VoL 20, No. 5 (pp.296-301)
B A I C A L E I N I N H I B I T E D M D R l VIA N F - K B P A T H W A Y I N B R E A S T C A N C E R M C F - 7 / A D R C E L L S
Tran TltlHien''^*
'Thaibinh Medical University, Vietnam; ^Faculty of Medicine, Lund University. Sweden
*Corresponding author: [email protected] (Received March, 17*, 2015)
Summary
Baicalein Inhibited MDRl Via NF-KB Pathway in Breast Cancer MCF-7/adr Cells P-glycoprotein (P-gp) accounts for most intrinsic and acquired cancer multidrug resistance (MDRl) and its inhibition is an effective way to reverse cancer drug resistance. Baicalein, a natural isoflavonoid compound suppressed MDRl expression in MCF-7/adriamycin (MCF-7/adr), a human breast MDR cancer cell line. Baicalein treatment significantly inhibited MDRl expression, mRNA transcription, as well as MDRl promoter activity in MCF-7/adr cells. In addition, the suppression ofP- glycoprotein was accompanied by partial recovery of intracellular drug accumulation, leading to increased toxicity of adriamycin and fluorescence of rhodamine 123, indicating that baicalein reversed the MDR phenolype by inhibiting the dmg efflux function of MDRl. In addition, NF-KB activity was decreased by baicalein. Taken together, these results suggested that baicalein down-regulated MDRl expression via NF-KB activity in breast cancer MCF-7/adr cells.
Keywords: Baicalein. MDRl. NF-KB. MCF-7/adr cells.
1. Introduction outside. In tumor cells, over-expressed P-gp is Cancer cells can develop resistance to responsible for the efflux of various structurally structurally diverse and mechanistically unrelated and functionally related or unrelated hydrophobic anticancer drugs, a phenomenon termed muhidrug chemotherapeutic agents from cells, which resistance (MDR) [1]. P-glycoprotein (P-gp), results in a low efficacy of chemotherapy [3]. P- encoded by the MDRl gene, is a multidrug gp demonstrates a broad spectrum of substrate transporter that functions as an adenosine specificities toward vinca alkaloids, andiracyclines, triphosphate (ATP)-dependent drug efflux pump taxens, and epipodophylotoxins and is responsible possessing broad substrate specificity [2]. P-gp is for the intrinsic and acquired drug resistance in understood to act as a pump that detects and numerous human cancers. P-gp-mediated drug removes its substrate from inside the cells to resistance can be effectively overcome by either
