The resurgence of malaria due to drug-resistant strains of the parasite and insecticide-resistant strains of the mosquito vector retains the attention of many scholars. As a result, there have been various efforts to combat the problem of parasite resistance, such as reversing chloroquine resistance, using combination therapy, and discovering new antimalarial compounds from various sources, especially from traditional medicinal plants [
Medicinal plants have contributed significantly to current malaria treatment. Artemisinin and quinine, main antimalarial drugs today, are provided by medicinal plants, but artemisinin-resistant parasite strains are emerging [
There is much research on traditional herbal medicines in Africa, but very little of it is clinical. In a review, out of more than 1200 plant species reportedly used for the treatment of malaria, only 13 have undergone clinical trials, although hundreds have been tested in the laboratory [
Furthermore, in Africa, there are many herbal remedies available on the market but with no clinical trials establishing their safety and efficacy. This situation is due to the insufficient implementation of regulation on medicines and especially on phytomedicines, which is quite common in many developing countries. In order to fill this gap, leading to standardized herbal medicine which will be officially authorized on the market, a “reverse pharmacology” approach is being used, where clinical evaluation is prioritized. The primary objective of this approach is to improve the utilization of herbal medicines, which are already in use and in the process of being approved [
In Benin, many antimalarial herbal medicines have been developed and are widely used by the population. Clinical information was collected retrospectively on those remedies. In a survey conducted by Dr. Allabi,
The purpose of phase I clinical trial was to establish
Commercial presentation of
Phase I clinical trial was a simple, one-arm prospective evaluation of safety and tolerability in ten (10) apparently healthy male volunteers aged 18 to 40, with negative or positive parasitaemia. They were selected using the Lot Quality Assurance Sampling (LQAS) method and were enrolled if they complied with a number of inclusion and exclusion criteria. The recruitment of volunteers has been among the population of Abomey-Calavi; all have given a written consent after being informed. The information sheet and the consent form were both in conformity with the International Guidelines for Research Involving Human Subjects of the WHO, and the Declaration of Helsinki on medical research involving human subjects. After obtaining the signed consent forms, the volunteers were taken for further examinations to determine the baseline of their biological parameters. Safety and tolerability were clinically monitored by comparison of vital signs, physical characteristics, and haematological and biochemical parameters at days 0, 1, 2, 3, 4, 5, 7, and 14, to the respective reported data. Biochemical parameters were determined using the CHEM-7 Cerba® Mannheim spectrometer and the automatic haematology analyzer HORIBA® ABX Micros ES60 was used for haematological parameters.
The trial was conducted from February 2017 to April 2017 at the Teaching Hospital Center of Abomey-Calavi/Sô-Ava in the Department of Internal Medicine and the evaluation of haematological and biochemical parameters as well as parasitaemia was carried out in the biomedical analysis laboratory of this hospital.
These are being male, aged between 18 and 40, being apparently healthy (this means that those subjects were not feverish and had no other signs of malaria with positive or negative parasitaemia) as observed by physical examination and haematological and biochemical investigations, no chronic pathology, no gastrointestinal intolerance including nausea, vomiting, and/or diarrhea, absence of any complaints before the treatment, no allergy to the constituents of
These include development of severe malaria according to WHO definition during the study, having an evidence of significant clinical abnormalities detected by a physician after inclusion, any kind of medical or herbal treatment intake during the study without the investigator consent, and alcohol intake during the study.
35 mL of
This dose corresponded to 42.86 mg/kg body weight, which is significantly lower than the nontoxic 2000 mg/kg dose used during the preclinical studies [
Clinically, the safety of the product was assessed through the vital signs such as blood pressure, pulse and respiratory rate. The adverse events were documented daily using a checklist [
Tolerability was investigated through the monitoring of biological parameters using blood and urine samples at days 1, 2, 3, 4, 5, and 7. The main criteria to evaluate biological tolerability of CoBaT-Y017 were based on blood chemical which includes hepatic function test such as ASAT (12 to 42 IU/L) and ALAT (10 to 48 IU/L), renal function tests such as creatinine considered normal, when the value was between 7 and 13 mg/L, and haemoglobinuria and proteinuria whose absence in the urine were considered as normal. The glycaemia (Gly) was also monitored in which normal levels ranged from 3.3 to 6.1mmol/L as well as the prothrombin (TP) rate in which normal value is at least 72%. For complete blood numeration, the haematological parameters were considered normal when haemoglobin rate (Hb) is 13 to 17 g/dl, red cell number (NR) is 4.2 to 5.7 (X106 per mm3), hematocrit rate is 40 to 50%, mean corpuscular volume is 85 to 95 fl, mean corpuscular concentration in haemoglobin is 32 to 36%, platelet number (Plat) is 150 to 400 (X103 per mm3), leukocyte number (NW) is 4 to 10 (X103 per mm3), neutrophil rate is 40 to 75%, lymphocyte rate is 20 to 45%, eosinophil rate is 1 to 4%, basophil rate is 0 to 1%, and monocyte rate is 2 to 8%.
All parameters were compared to the baseline values [
Data treatment forms were created to collect clinical and biological data for each volunteer. The data collection forms included the clinical follow-up and the biological follow-up. At the end of the data collection, all data of each participant was brought together in case record forms.
The data was entered in Microsoft Office Excel 2013 and transferred to Stata 11 and SPSS 16.0 for further analysis. The data was expressed as mean ± standard deviation and/or standard error when needed. Comparison of different parameter means over time was carried out using Friedman
This study protocol was approved by the Research Ethics Committee of Applied Biomedical Sciences Institute (CER-ISBA) of Cotonou at this reference: N°103 du 09/01/17. The clinical trial was conducted in compliance with the Declaration of Helsinki on medical research involving human subjects, the Good Clinical Practice (GCP) guidelines, and the Standard Operating Procedures elaborated by the UNDP/World Bank/WHO/TDR [
Ten volunteers were recruited to participate in this phase I clinical trial study, based on the inclusion and noninclusion criteria.
The median age of participants was 25 years with extremes of 22 and 35 years and only men in this age group were enrolled. The average body mass index (BMI) of these volunteers was 19.91 ± 2.01 kg / m2. This was a normal BMI value and corresponded to subjects of normal weight. Only one participant had a BMI below 18 kg/m2 and was considered as a lean subject. However, the physical examination of this volunteer, as for the others, did not show any abnormalities or pathological features.
At inclusion, no abnormalities were observed for the pulse of volunteers whose mean at baseline was 66.00 ± 3.06 beats per minute. The mean systolic blood pressure (SBP) of subjects at baseline was 12.00 ± 0.67 cmHg and the mean diastolic blood pressure (DBP) was 7.00 ± 0.94 cmHg. As these values are within the normal limits, all volunteers therefore had normal blood pressure at inclusion. These volunteers also had normal temperatures; mean at baseline was 36.4 ± 0.7°C.
Overall, the participants’ haematological and biochemical parameters were normal at inclusion. The haemoglobinuria and proteinuria tests were also negative for all volunteers at inclusion, thus showing that none of the volunteers suffered renal or hepatic disease.
20% of volunteers found
The pulse values of the study subjects remained within normal limits during the monitoring period. Nonsignificant changes were observed between consecutive days after administration and until day 14 (Figure
Pulse evolution during the monitoring period.
Systolic and diastolic blood pressures remained normal throughout the monitoring period and did not show significant variation.
Figure
Systolic blood pressure (SBP) evolution during the monitoring period.
Diastolic blood pressure (DBP) evolution during the monitoring period.
Figure
We observed nonsignificant variation of temperature throughout the monitoring period. The participants’ temperature remained between 36°C and 37°C until day 7 after the start of the treatment (Figure
Evolution of volunteers’ temperature during the monitoring period.
Table
Effects of
Haematological parameters | Inclusion D0 | Product Administration Days | Observation Days | Pvalue | ||||
---|---|---|---|---|---|---|---|---|
D1 | D2 | D3 | D4 | D5 | D7 | |||
Hb | 14.2 ± 0.6 | 14.2 ± 0.6 | 14.0 ± 0.7 | 14.1 ± 0.9 | 14.1 ± 0.8 | 14.1 ± 0.7 | 14.0 ± 0.8 | 0.46 |
NRx109 | 4.9 ± 1.8 | 5.1 ± 0.6 | 5.1 ± 0.5 | 5.2 ± 0.7 | 5.1 ± 0.6 | 5.0 ± 0.6 | 5.0 ± 0.7 | 0.50 |
NWx103 | 4.9 ± 0.9 | 4.9 ± 0.6 | 5.0 ± 1.3 | 4.9 ± 0.6 | 5.0 ± 0.7 | 4.9 ± 1.3 | 4.8 ± 0.7 | 0.87 |
Plat.x105 | 2.0 ± 0.4 | 2.4 ± 0.6 | 2.2 ± 0.6 | 2.1 ± 0.6 | 2.2 ± 0.6 | 2.2 ± 0.6 | 2.1 ± 0.5 | 0.18 |
TP | 74.8 ± 10.8 | 72.4 ± 10.7 | 73.4 ± 11.0 | 73.4 ± 9.5 | 70.0 ± 12.0 | 77.1 ± 12.0 | 73.7 ± 13.7 | 0.23 |
Legend: D: day; Hb: haemoglobin; NR= number of Red cells; NW= number of white cells; Plat: platelet; TP: prothrombin rate.
Table
Effects of
biochemical parameters | Inclusion | Product Administration Days | Observation Days | p value | ||||
---|---|---|---|---|---|---|---|---|
D1 | D2 | D3 | D4 | D 5 | D 7 | |||
CREA | 10.4 ± 1.6 | 10.4 ± 1.6 | 10.0 ± 2.1 | 10.1 ± 3.3 | 10.5 ± 2.1 | 10.5 ± 2.2 | 10.1 ± 1.8 | 0.46 |
TGO | 24.5 ± 6.1 | 26.1 ± 7.8 | 26.5 ± 5.2 | 28.5 ± 7.3 | 27.6 ± 11.4 | 30.0 ± 22.0 | 29.2 ± 18.5 | 0.56 |
TGP | 17.8 ± 5.3 | 18.5 ± 5.6 | 21.0 ± 11.4 | 22.0 ± 6.2 | 21.1 ± 6.3 | 22.5 ± 7.8 | 19.6 ± 11.2 | 0.87 |
GLY | 0.8 ± 0.1 | 0.8 ± 0.1 | 0.8 ± 0.1 | 0.8 ± 0.1 | 0.7 ± 0.1 | 0.7 ± 0.1 | 0.8 ± 0.1 | 0.28 |
Legend: D: day; CREA: creatinine; TGO: transaminase GO; TGP: transaminase GP; GLY: glycaemia.
Daily haemoglobinuria and proteinuria tests remained negative during the monitoring period.
According to these results,
Table
Activity of
patients | Inclusion | | Observation Days | |||||
---|---|---|---|---|---|---|---|---|
D1 | D2 | D3 | D4 | D5 | D7 | D14 | ||
P1 | - | - | - | - | - | - | - | - |
P2 | - | - | - | - | - | - | - | - |
P3 | - | - | - | - | - | - | - | - |
P4 | +8700 p/ | - | - | - | - | - | - | - |
P5 | - | - | - | - | - | - | - | - |
P6 | - | - | - | - | - | - | - | - |
P7 | - | - | - | - | - | - | - | - |
P8 | - | - | - | - | - | - | - | - |
P9 | +380p/ | - | - | - | - | - | - | - |
P10 | - | - | - | - | - | - | - | - |
Legend: P: participant; D: day; +: presence of trophozoites of
During the treatment, 60% (six out ten) of volunteers said they felt no adverse events, while 20% (two out of ten) reported both increased appetite and drowsiness, and another 20% reported increased appetite only. However, all adverse events disappeared at once with the end of the treatment (day 5).
According to the classification of adverse events of CTCAE, the two events recorded (increased appetite and drowsiness) are categorized as grade 1 adverse events.
Our phase I study fills part of the scientific gap [
Results of this study revealed no variation of hepatic transaminases profile over the normal limits after administration of the product. This result is similar to what was observed with another antimalarial herbal medicine, the extract of
The absence of serious adverse events corroborated the results obtained in a study on a plant whose extract is one of the compounds of our investigated product [
The main interest of this work was that we started from the realities of field, remedy used informally and without marketing authorization, to appreciate the safety and the tolerability of use of the
Our results demonstrate no significant adverse events and toxicity of
Body mass index
Research ethic committee of apply biomedical sciences institute
Coopération Pharmaceutique Biologique et Technique
Creatinine
Common terminology criteria for adverse events
Diastolic blood pressure
Glutamic oxaloacetic transaminase
Glutamic pyruvic transaminase
Glycaemia
Haemoglobin
Day
Lot Quality Assurance of Sampling
Number of red cells
Number of white cells
Systolic blood pressure
Prothrombin rate.
The data used to support the findings of this study are available from the corresponding author upon request.
This research was partially supported by COPHARBIOTEC Laboratory.
The authors are grateful to the volunteers who participated in this study.