EFFECTS OF ARTENOLANE MALEATE-PIPERAQUINE PHOSPHATE

FIXED DOSE COMBINATION (AM-PFFDC) ATIMALARAIAL DRUG ON

ANTIOXIDANT PARAMETERS OF WISTAR RAT

BY

IHIONU ONYEKACHI DAVID

2017294037

A RESEARCH PROJECT SUBMITTED

TO

DEPARTMENT OF PHARMACOLOGY AND TOXICOLOGY

FACULTY OF PHARMACEUTICAL SCIENCES

CHUKWUEMEKA ODUMEGWU OJUKWU UNIVERSITY,

IGBARIAM

IN PARTIAL FULFILMENT FOR THE AWARD OF BACHELOR OF

PHARMACY (B.PHARM)

SUPERVISOR: DR. EARNESTO.ERHIRHIE

NOVEMBER, 2023

i
 DECLARATION

I do hereby declare that this research project “Effects of Artenolane Maleate-Piperaquine

Phosphate Fixed Dose Combination (AM-PFFDC) antimalarial drug on antioxidant parameters

of wistar rat” was carried out by me, under the supervision of “Dr. Earnest O. Erhirhie”. This

project work is original and has not been carried out by anyone. It has never been submitted in

part or full for other degree or diploma in the University or other Institutions.

……………………………

Ihionu Onyekachi David

Date: November, 2023

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 APPROVAL

This research, “Effects of Artenolane Maleate-Piperaquine Phosphate Fixed Dose Combination

(AM-PFFDC) antimalarial drug on antioxidant parameters of Wistar rat” has been and approved

as having met the requirements of the Department of Pharmacology and Toxicology, Faculty of

Pharmaceutical Sciences, Chukwuemeka Odumegwu Ojukwu University, Igbariam for award of

Bachelor of Pharmacy (B. Pharm).

……………………….. ………………………..

Dr. Earnest O. Erhirhie Date

(Project supervisor)

………………………. ………………………

Dr. Earnest O. Erhirhie Date

(Head of Department)

................................. ………………….

Prof. T.C. Akunne Date

(Dean of the Faculty)

……………………. ……………………

(External Supervisor) Date

iii
 DEDICATION

This work is Firstly dedicated to God Almighty and the cherished family of Late Sir Raymond

Ihionu, whose invaluable love, care, unwavering support, and encouragement have been the

guiding force fueling my journey in the pursuit of knowledge.

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 ACKNOWLEDGEMENT

Firstly, I am grateful to God Almighty for his infinite kindness, granting me the zeal and wisdom

to articulate ideas, leading to the resounding success of this work. Of great importance, I say a

big thank you to my renowned supervisor and H.O.D., Department of Pharmacology and

Toxicology, Dr. Earnest O. Erhirhie for his attention, patience and time.

Additionally, heartfelt gratitude is extended to the Staff of the Department and the Dean of the

Faculty of Pharmaceutical Science at Chukwuemeka Odumegwu Ojukwu University, Anambra,

for facilitating the execution of this work within the Department and the Faculty’s animal

facility. To all our esteemed lecturers, I wholeheartedly appreciate and thank you for imparting

knowledge and graciously sharing wisdom from your wealth of experience.

My profound gratitude to my parents, and all my family members in persons of Late Sir

Raymond Ihionu and all my siblings especially Lady Mrs Benedicta Azubuike, Late Mrs

Augusta Nnabuihe and Miss Monica Ihionu for their support, prayers and encouragements

throughout my stay in the university.

A special tribute is dedicated to my course mates, with whom I endured sleepless nights and

relentless coursework, overcoming challenges together. Gratitude is also extended to my loyal

friends, Chioma, Chieloka and Princess for adding an element of enjoyment to our shared

journey through school.

Special gratitude and thanks go to all whose name has not been mentioned, whose effort,

support, prayers, best wishes and encouragements help me throughout my study.

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 ABSTRACT

Background and aim:

Antioxidants are crucial for protecting cells from damage caused by oxidative stress. As

Artenolane Maleate-Piperaquine Phosphate Fixed Dose Combination (AM-PFFDC) effectively

fights malaria, this research aims to uncover whether it might disrupt the body's antioxidant

defense mechanisms. This study examines the potential impact of AM-PFFDC, a promising

antimalarial drug, on the body's antioxidant parameters.

Method:

Acute toxicity test was carried out using Lorke’s method. Twenty-seven rats were divided into

five groups and received AM-PFFDC at doses of 0, 11.625, 23.25, 46.5, and 93 mg/kg for four

consecutive days. Body weight, food, and water intake were monitored each day, and blood

samples were collected on the fifth day of the experiment to measure the levels of superoxide

dismutase, malondialdehyde, Catalase and Glutathione Reductase in the blood. The data were

analyzed using one-way ANOVA, where the significance level was set at 0.05.

Result:

In an acute toxicity test using Wistar rats, AM-PFFDC showed a high LD 50 of 3808 mg/kg. At 93

mg/kg, adverse effects included reduced feed and water intake. Despite increased Glutathione

(GSH) levels in all treatment groups compared to controls, there were no significant changes in

Superoxide Dismutase, Catalase, or Malondialdehyde levels with various AM-PFFDC doses.

AM-PFFDC had no significant impact on body weight.

Conclusion:

AM-PFFDC may not affect antioxidant parameters, body weight, feed intake, and water intake

depending on dosage and Physiology.

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 TABLE OF CONTENTS

Title page……………………………………………………………………………………i

Declaration………………………………………………………………………………….ii

Approval.………………………………………………………………………………........ii

Dedication………………….……………………………………………………………….iii

Acknowledgement…………………………………………………………………………..iv

Abstract………………………………………………………………………………………v

Table of content……………………………………………………………………….……..vi

List of tables………………………………………………………………………………….x

List of figures…………………………………………………………………………….......xi

CHAPTER ONE: INTRODUCTION AND LITERATURE REVIEW

1.1 Introduction to the study ……………………………………………………………………...1

1.2 Statement of Problem …………………………………………………………………………2

1.3 Significance of the Study ……………………………………………………………………..2

1.4 Justification of the Study ……………………………………………………………………..3

1.5 Aim and specific objectives of the study …………………………………………………......3

1.6 Scope and Limitations …………………………………………………………………….......4

1.7 Literature Review ……………………………………………………………………………..4

1.7.1 Malaria and its Treatment …………………………………………………………………..5

1.7.2 AM-PFFDC Antimalarial Drug………………………………………………………….....5

1.7.3: Mechanism and Utilization of AM-PFFDC………………………………………………...6

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1.7.4 Antioxidant Parameters and Their Importance ……………………………………………8

1.7.5 Previous studies on Evaluation of Some Anti-Malaria Drugs on Anti-oxidant properties in

Rats
………………………………………………………………………………………………….10

1.7.5.1 Previous study on the Effects of Chloroquine Treatment on Antioxidant Enzymes in Rat
Liver and Kidney ……………………………………………………………………………....10

1.7.5.2 Study on the Effects of Artemisinin on Oxidative Stress Markers in Mouse Heart and
Lung Tissues in an Experimental Model of Epileptic Seizure ………………………………… 11

1.7.5.3 Study on the Influence of Coadministration of artemether and lumefantrine on selected

plasma biochemical and erythrocyte oxidative stress indices in female Wistar rats ……………12

1.7.5.4 Effects of primaquine and chloroquine on oxidative stress parameters in rats ………...13

1.7.5.5 Antimalarial therapy and changes in oxidative stress indices in falciparum malaria
infection in Human subjects ……………………………………………………………………..14

1.7.5.6 Investigation of the Potential Impact of AM-PFFDC on the Antioxidant Properties in

Wistar Rats ………………………………………………………………………………………15

CHAPTER TWO: MATERIALS AND METHODS

2.0 Materials and Methods ………………………………………………………………………16

2.1 Materials…………………………………………………………………………………….16

2.1.1 Apparatus ………………………………………………………………………………….16

2.1.2 Equipment …………………………………………………………………………………16

2.1.2 Other Materials ……………………………………………………………………………17

2.2 Method ………………………………………………………………………………………17

2.2.1 Experimental Animals …………………………………………………………………….17

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2.2.2 Experimental Design ……………………………………………………………………..18

2.2.3 Dosage Calculation………………………………………………………………………..19

2.2.4 Treatment………………………………………………………………………………….19

2.2.5 Body Weight, Feed, and Water Intake Determination…………………………………….19

2.2.6 Sample Collection………………………………………………………………………. ...20

2.2.7 Laboratory Analysis……………………………………………………………………......21

2.2.7.1 Determination of Superoxide Dismutase………………………………………………...21

2.2.7.2 Determination of Malondialdehyde……………………………………………………...21

2.2.7.3 Determination of Catalase………………………………………………………………..22

2.2.7.4 Determination of Glutathione Reductase………………………………………………...23

2.2.8 Method of Data Analysis…………………………………………………………………..23

CHAPTER THREE: RESULTS

3.1Acute toxicity test result ……………………………………………………………………..24

3.2Effects of AM-PFFDC Antimalarial Drug on body weight of Wistar rats…………………...25

3.3 Effect of AM-PFFDC Antimalarial Drug on feed intake assessment ……………………….26

3.4 Effect of AM-PFFDC Antimalarial Drug on water intake assessment …...…………………27

3.5 Effect of AM-PFFDC Antimalarial Drug on antioxidant parameters ………………………28

CHAPTER FOUR: DISCUSSION AND CONCLUSION

4.1Discussion ……………………………………………………………………………………29

4.2 Conclusion ………………………………………………………………………………..…33

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4.3 Recommendation ……………………………………………………………………………34

References ………………………………………………………………………………………37

Appendix

Appendix 1: List of abbreviations …………………………………………………………….…43

Appendix 2: Certificate of ethical approval ……………………………………………………..44

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 LIST OF TABLES

Table 1: Acute toxicity test result…………………………………………………………….26

Table 2:Effects of AM-PFFDC Antimalarial Drug on body weight of Wistar rats ……………27

Table 3:Effect of AM-PFFDC Antimalarial Drug on feed intake assessment …………………28

Table 4: Effect of AM-PFFDC Antimalarial Drug on water intake assessment ………………29

Table 5: Effect of AM-PFFDC Antimalarial Drug on antioxidant parameters ……………..30

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 LIST OF FIGURES

Figure 1: AM-PFFDC tablet………………………………………………………………………..8

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