Project Abstract

Abstract
Malaria remains a major public health concern globally, with millions of cases reported annually. The emergence of drug-resistant strains of Plasmodium parasites has led to a need for new and effective anti-malarial treatments. Traditional medicine has been a potential source of novel anti-malarial compounds, and Azadirachta indica, commonly known as neem or dogoyaro, has been used in various traditional medicinal practices for its anti-microbial properties. This study aimed to evaluate the chemical components of Dogoyaro liquid extract and assess its potential as an anti-malarial agent. Azadirachta indica leaves were collected and extracted using a solvent extraction method to obtain the liquid extract. The extract was then subjected to phytochemical screening to identify the presence of bioactive compounds such as alkaloids, flavonoids, tannins, saponins, and terpenoids, which are known for their anti-malarial properties. The chemical composition of the extract was further analyzed using chromatographic techniques such as high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) to identify specific compounds present in the extract. The anti-malarial activity of the Dogoyaro liquid extract was evaluated using in vitro assays against Plasmodium falciparum, the parasite responsible for causing malaria in humans. The extract was tested at various concentrations to determine its inhibitory effects on parasite growth. Additionally, the cytotoxicity of the extract was assessed using mammalian cell lines to ensure its safety for potential therapeutic use. The results of the phytochemical screening revealed the presence of alkaloids, flavonoids, tannins, and terpenoids in the Dogoyaro liquid extract, which are compounds known for their anti-malarial properties. The chromatographic analysis identified specific compounds such as azadirachtin, nimbin, and nimbidin, which have shown anti-malarial activity in previous studies. The in vitro assays demonstrated that the Dogoyaro liquid extract exhibited dose-dependent anti-malarial activity against Plasmodium falciparum while maintaining low cytotoxicity to mammalian cells. In conclusion, the chemical evaluation of the Dogoyaro liquid extract revealed the presence of bioactive compounds with potential anti-malarial properties. Further studies are warranted to elucidate the mechanism of action of these compounds and evaluate the extract's efficacy in animal models to support its development as a novel anti-malarial treatment.

Project Overview

INTRODUCTION
Malaria infections are characterized by defective immune responses with poor efficacy against infection, and in some cases, with immunopathology. Severe malaria is a complex multisystem disorder. Complications such as cerebral malaria, anemia, acidosis, jaundice, respiratory distress, renal insufficiency, coagulation anomalies and hyperparasitemia can occur in severe malaria[2]. About 300-500 million clinical cases and 1.2?2.8 million deaths due to malaria occur each year[3,4]. The rising problem of resistance to the classical drugs (chloroquine and sulphadoxine pyrimethamine)[5?7] and the problem of recrudescence of malaria after treatment with artemisinin stress the need for new antimalarial agents[8].
The malaria drugs quinine and artemisinin are both plant derivatives and are obtained from Cinchona species[9,10] and Artemisia annua[11,12] respectively, suggesting that other effective malaria drugs might be plant-derived. Azadirachta indica plants from the Meliaceae family are extensively used as traditional remedies against malaria in the tropics[9,13,14]. Several studies demonstrated that A. indica leaf, seed and stem bark extracts possess in vitro inhibitory activity on Plasmodium falciparum asexual stages[15?17]. In vitro screening of purified limonoids from Neem revealed that gedunin and nimbolide are the most active molecules against P. falciparum.
MacKinnon et al. examined 60 extracts from 22 species of Meliaceae by characterizing their in vitro antiplasmodial activity against chloroquine-sensitive (D6) and chloroquine-resistant (W2) malaria clones (Dd2, Indochina)[15,19]. Twelve extracts were found to have activity against P. falciparum, especially extracts of Cedrela odorata wood and Azadirachta indica leaves, both containing the limonoid, gedunin[20]. Gedunin has been repeatedly re-isolated as an active antimalarial principle, most recently from Khaya grandifoliola[21]. The antimalarial properties of several plant extracts have been studied in mice[22]. Therefore, we sought to examine the anti-plasmodial activity of Azadirachta indica, commonly known as Neem, a plant used in Sudan, India and other countries as a folk medicine, in a human malaria investigation of malaria produced in Plasmodium berghei ANKA infected mice.
To examine the effects of an antimalarial treatment, we measured physiological and morphological responses. We also examined cerebral malaria. Neuronal ischemia and inflammation from malaria, depending on the acuteness and intensity of the symptoms, can lead to cellular necrosis or apoptosis[23]. Caspases play important roles in the commitment and execution phases of apoptosis[24]. Importantly, caspase-3 acts as a major downstream effector mediating neuronal apoptosis. We examined protection from caspase-dependent apoptosis as a mechanism of malaria treatment effectiveness.
1.1 AIM OF PROJECT
Many researches have been carried out on neem plant (Azadirachta indica) and results have shown that it has both medicinal and pharmacological properties against malaria. However, there are no documented information relating the mineral properties inherent in the leaf of the plant. Consequently, this work was aimed at determining some chemical component present in the water extract of neem leaf using the modern atomic absorption spectrophotometric analysis.

1.2 SCOPE OF THE STUDY
Extracts from neem tree (Azadirachta indica A. Juss) also called ?dogonyaro? in Nigeria are most consistently recommended in ancient medical texts for gastrointestinal upsets, diarrhea and intestinal infections, skin ulcers and infections, and malaria (Schmutterer,1995). India encouraged scientific investigations of neem tree as part of his program to revitalize Indian tradition and also increase commercial interest on neem (Stix, 1992) and presently some authors believe that no other plant or tree in the world has been so extensively researched or used, in all possible capacities so far. In Africa extracts from Neem leaves have provided various medicinal preparations (Ekanem, 1971 and Udeinya, 1993).
SIGNIFICANCE OF THE STUDY
Development of modern non-toxic drugs from neem has earlier been suggested (Biswas et al, 2002) and IRAB represents such dream. This fractionated acetone/water neem leaf extract is the only drug reported to have activities against Malaria. Recent in-vivo studies have substantiated the earlier in-vitro reports of pharmacological properties.
OBJECTIVE OF THE STUDY
Main Objective
1. Evaluation of the chemical component of dogoyaro liquid (azadirachta indica) for anti-malaria

Specific Objective
1. To examine the chemical component of dogoyaro leaf which serve as drug to many illness especially Malaria