1. Stability of Antimalarial Preparations Rutendo Kuwana Accra, December 2009

2. 2 |2 | PQ: Artemisin-derivative issues No innovator FPP registered in the ICH region. Therefore No comparator available for: –Pharmaceutical equivalence studies –Bioequivalence studies The APIs and FPPs not official in the internationally used major pharmacopoeias WHO guides/SOPs apply to multisource FPPs. ICH guides therefore used

3. 3 |3 | Expression of Interest – oral dosage forms Artesunate* + Amodiaquine Artemether* + Lumefantrine* Artesunate* + Mefloquine Artesunate* + SP (sulphadoxine / pyrimethamine) Dihydroartemisin+Piperaquine Phosphate* * No comparator at the beginning* High quality-risk API +... FDC or co-blistered (co-packaged) FPPs * No comparator to date

4. 4 |4 | EOI – other dosage forms Artemether Injection and rectal FPPs Artemotil (arteether) Injection Artesunate Injection and rectal FPPs Only FPPs listed in the EOI will be discussed.

5. 5 |5 | Availability of monographs for drug substances - International Pharmacopoeia Artemether Artemisinin Artemotil Artenimol Artesunate Mefloquine Hydrochloride Proguanil Hydrochloride – Also in BP Amodiaquine and Amodiaquine Hydrochloride – Also in USP

6. 6 |6 | International quality standards Lumefantrine Pyrimethamine BP, PhEur, PhInt, USP Sulphadoxine BP, PhEur, PhInt, USP Piperaquine Dihydroartemisinin

7. 7 |7 | Other Antimalarial API –APIs described in monographs of major international pharmacopoeias (  10 years) Chloroquine, Dapsone, Quinine, Mefloquine, Trimethoprim –APIs not described in monographs of major international pharmacopoeias Chlorproguanil,, Naphthoquine, Pyronaridine

8. 8 |8 | Properties of Artemisinin derivatives Artemisinin (C 15 H 22 O 5 ) –7 centres of asymmetry –2 7 potential isomers –One isomer in biosynthesis –Chemical synthesis Feasible Economically unacceptable Chemical derivatization at C-10 (carbonyl-moiety) converts C-10 into an additional stereoisomeric center:  - and  -isomers are formed 1 2 3 4 5 6 5a 8a 7 8 9 10 11 12 12a

9. 9 |9 | Artemisinin Active antimalarial constituent of the traditional Chinese medicinal herb Artemisia annua L., Compositae Although Artemisinin has seven (7) centers of assymetry Artemisia annua makes only one configuration Practically insoluble in water The bond energy of the O-O bond is ~30 kcal/mol When the peroxide comes into contact with high iron concentrations, the molecule becomes unstable and "explodes" into free radicals. The API, the capsules and the tablets are official in the Ph. Int. Not included in the current EOI.

10. 10 | Artesunate Very slightly soluble in water The ester linkage is in alpha configuration. Both the API and the tablets are official in the Ph. Int. Two functional groups are liable to decomposition

11. 11 | Metabolism of Artemether and Artesunate

12. 12 | Mefloquine hydrochloride Has an optically active carbon Very slightly soluble in water Has no reactive functional groups under general environmental conditions

13. 13 | Pharmaceutical information Artemisinin derivatives may have α- or β-configuration and each of them can exist in two conformations. The literature does not reveal any impact of the geometric isomerism on efficacy, safety or quality of artemisinins. The internal peroxide bound is the most reactive part of the molecule. When the peroxide comes into contact with high iron concentrations, the molecule becomes unstable and "explodes" into free radicals. The ester bond of artesunate is liable to hydrolysis. The non-artemisinin APIs in the EoI are chemically stable.

14. 14 | Potential impurities of Artemisinins Starting material (extracted from herbal sources) Starting materials from vegetable origin should be fully characterized and a contaminant profile should be established

15. 15 | Potential impurities of Artemisinins II Impurities contained in the "starting material" Artemisinin –Biosynthetic by-products Arteannuin B, Artemisitene, Artemisinic acid, –Extraction from fresh leaves with CHCl 3 Thujone (?) –Cultivation reagents Pesticide residues, fumigants, mycotoxins –Solvents from the extraction process Hexane, benzene, acetonitril, ether, pentane, chloroforme…..(?) diesel, fuel (?) [ICH Q3A (R)]

16. 16 | Potential impurities of Artemisinins III Unreacted starting material –Artemisinin (starting material for derivatives) –Artemisinic acid (starting material for dihydroartemisinin) –Dihydroartemisinin (starting material for derivatives) –…. Unreacted intermediates, by-products –  -Arthemether,  -Artheether –  /  -Dihydroartemisinin –  -Artesunate

17. 17 | Potential impurities of Artemisinins IV Reagents, catalysts, residual solvents –Methanol, acetonitril, chloroforme, acetone … –NaBH 4, succinic acid/anhydride, triethylamine, dimethylaminopyridine Degradants –Stability of ester-derivative (Artesunate) ether-derivative (Artemether, Arteether) lactone (Artemisinin) –Stability of artenimol (oxidation) –Susceptibility of endoperoxide bond to reduction Deoxyartemisinine (loss of active principle)

18. 18 | Compatibility of the API with excipients and diluents Select innovator excipients ( WHOPAR, EPAR ) Magnesium stearate is incompatible with salts of weak bases and strong acids (e.g. Amodiaquine.2HCl) because the formed MgCl 2 is highly hygroscopic and, as a result, its lubricant properties also change. The compatibility and in-use stability of the FPP with reconstitution diluents should be addressed, e.g. in Artesunate injection.

19. 19 | Increase in concentration of API During some stability studies of Artesunate, the assay results were increasing. The hydrolysis may yield artenimol and succinic acid. The latter can justify the increase in assay. The assay method may be considered to be "stability indicating” but not specific. +

20. 20 | Stress Testing Artesunate The drug substance degrades readily at heat conditions in extreme of 100ºC, it is also unstable under light conditions and both acid and base conditions by hydrolysis. The α-epimer of dihydroartemesinin is the major degradant under light conditions. α-Artemether is the major degradant under heat conditions in extreme of 100ºC although both the α and ß epimers of dihydroartemesinin are also significant degradants under this condition.