ANTIMALARIAL DRUGS

INTRODUCTION TO MALARIA
Malaria is one of the most common infectious diseases and an enormous public health problem. There are five species of the plasmodium parasite that can infect humans, These are : Plasmodium falciparum (Most Fatal) Plasmodium vivax (Mild Fatal) Plasmodium ovale (Mild Fatal) Plasmodium malariae (Mild Fatal) and Plasmodium knowlesi: causes malaria in macaques ( a type of monkey ) but can also infect humans. Although all of the species may cause significant illness, Plasmodium falciparum is responsible for the majority of serious complications and deaths

H U M A N

M A L.

P A R A S I T E S

HISTORY
In India the National Malaria Eradication Programme (NMEP) , started in 1958. There was a target for complete disappearance of this disease upto 1960s. However to the development of insecticidal resistance among mosquitoes and other factors, it was conceding that eradication of malaria is not possible. Therefore NMEP has been renamed National Antimalarial Programme. In 2001 NAMP has reported > 2 million malaria cases, out of which approx. 48% were due to Plasmodium falciparum with 1003 confirmed deaths. WHO estimates that actual number of malaria cases in India is 6 times more, i.e. 12-15 million.

THE LIFE CYCLE OF MALARIAL PARASITE

PATHOGENESIS OF MALARIA

CLASSIFICATION OF ANTIMALARIAL DRUGS

1. 4-Aminoquinolines : Chloroquine,Amodiaquine 2. Quinoline-methanol : Mefloquine. 3. Cinchona alkaloid : Quinine, Quinidine 4. Biguanides : Proguanil(Chloroguanide) 5. Diaminopyrimidines : Pyrimethamine 6. 8-Aminoquinoline : Primaquine, Bulaquine 7. Sulfonamides and sulfone :Dapsone 8. Tetracyclines : Tetracycline, Doxycycline 9. Sesquiterpine lactones : Artesunate, 10. Naphthoquinone : Atovaquone 11. Mannich base : Pyronaridine

ANTIMALARIAL DRUGS AND THEIR SITE OF ACTIONS

CHLOROQUINE
The 4-aminoquinoline chloroquine dates from the 1940s. It is still widely used as a blood schizonticidal agent ,effective

against the erythrocytic forms of all four plasmodial species,

but it does not have any effect on sporozoites, hypnozoites or gametocytes.

COMPLICATIONS
1. 2. After oral doses for Acute attack of Malaria: Pruritis (primarily in Africans)sometimes with Urticaria. Nausea, vomiting ,Abdominal Pain, Anorexia. Blurring of vision Chronic use of high daily doses in Rheumatoid diseases:Discoloration of nail beds and mucus membranes Bleaching of hair & Alopecia Irreversible Ototoxicity , Retinopathymyopathy & Peripheral Neuropathy. It can exacerbate dermatitis produced by gold /phenylbutazone therapy. 3. Large I/M or Rapid I/V administration: Excessive hypotension. Respiratory & cardiac arrest.

TREATMENT OF P. VIVAX MALARIA: A FLOW CHART

HERBAL ANTIMALARIAL DRUGS

CINCHONA Synonym:Jesuits bark,Peruvian bark Biological source : Cinchona calisaya, C.officinalis Parts used : Barks Chemical constituents: Quinine,quinidine,cinchonine,cinchonidine Mechanism of action:

Uses : Antimalarial action bitter stomachics & antipyretic

ARTIMISIA Synonym: Wormwood,quinghao. Biological source : Artemisia annua. Parts used : Unexpanded flower heads. Chemical constituents: Artimisinin, artemether, artether, artemisinic acid Mechanism of action : The endoperoxide bridge in its molecule appears to interact with heme in the parasite. Iron mediated cleavage of the bridge releases a highly reactive free radical species that binds to membrane proteins,causes lipid peroxidation,damages endoplasmic reticulum, inhibits protein synthesis & altemately lysis of parasite. Uses :Artemisinin shows antimalarial effects by its rapid blood schizonticidal activity. Recent research : Artemisia is the most potent drug for treating cerebral malaria

SITE OF ACTION OF ARTIMISIA

PLUMBAGO BENESIS Biological source : Plumbago benesis. Family : Euphorbiaceae. Parts used : Flowers. Chemical constituents: Plumbagin , 3,3 plumbagin , 8,8 plumbagin. Uses :Have in-vitro activities against P.falciparum and epimastigotes of T.cruzi & also show antileishmaniasis activity.

TINOSPORA CARDIFOLIA (GILOYA) Family- Menispermaceae . Chemical constituents- The active glycoside is giloin . Uses It is used in jaundice,diabetes, gynacological & joint disorders. Also having antiviral,antibacterial, & hypoglycemic activity. The fine powder of dried stem prescribed (10gm BD for 5 days) for the treatment of malarial fever.

ANTIMALARIAL DRUG DISCOVERY PATH

ANTIMALARIAL SCREENING
IN VITRO SCREENING TESTS The test is carried in a 96 flat-bottomed wells in microtitre plates. Extract is dissolved or micronised in ethanol & a series of 6 conc. prepared by 10 fold dilutions in RPMI 1640 medium. The ethanol conc. of dilutions for testing is kept below 0.1%. Aliquots(50l) of culture medium is then added to each well. Human blood(50l) containing 1% parasitaemia is added to each well

Two series of controls are set up 1. with parasitised blood without addition of extract 2. with uninfected red blood cells. After incubation in 3% O2 ,4% CO2 & 93% N 2 gas phase for 18h at 37C,50l of G-[3 H] hypoxanthine is added to each well. Incubation continued at 37C for a further 18-24h. RBCs are harvested with a Titertek Cell Harvester washing out the wells with normal saline & filtering through glass fibre
membrane.

 Membranes are dried , placed in scintillation fluid & incorporation of [3H] hypoxanthine determined by scintillation counting. EVALUATION: The % inhibition of incorporation & IC50 values are determined.

INVIVO SCREENING TESTS

1. PETERS TEST Male mice(swiss albino) weighing 202 g are maintained at 22C in batches of 5 & fed on std diet. Blood from donor mouse with rising parasitaemia is diluted in tissue culture medium so that each 0.2ml contains 107 infected red cells. Each mouse receives 0.2ml i.v via the tail on day zero. Extract is either suspended or dissolved by trituration or sonication after the addition of 0.2% solution of Tween 80 or 0.5 DMSO.

 Initial aq. conc. of doses ranging b/w 1 and 100 mg/kg are administered daily from initial day of infection for 4 successive days either by s.c or oral routes. On the 5 th day samples are taken from tail blood & stained with a suitable stain(eg.Giemsa) EVALUATION: Parasitised red cells are recorded as a percentage of total ED50 values(i.e 50% suppression of parasites when compared with controls)

2.THE RANE TEST

The basis of the test is to compare the effect of a std inoculum of P.berghei,which kills mice within 6 days with extension of survival time to 12 days by a single dose of test compound. An inoculum of 106 infected donor cells is given i.p on day 1 . Test compound is suspended in arachis oil are given s.c at an initial dose range of 640,320,160 & 80 mg/kg on day 4. EVALUATION: The minimum effective dose(MED) is obtained & compared with the maximum tolerated dose(MTD) that produces no more than one in five toxic deaths.

CONCLUSION
..And

REFERENCES
KD Tripathi. Essentials of medical pharmacology;5th Edn. 2004,Jaypee brothers publication, New Delhi :587-598. Rang & Dale's, Pharmacology 7th edition. N.S Parmar & Shiv prakash. ,Screening Methods in Pharmacology H.Gerhard Vogel ,Drug Discovery And Evaluation,

Pharmacology Bioassay