SECTION III 

 RESULTS 



ADH-BSA Conjugates 



Synthesis 



Preliminary investigations of the targeting of a- 

 amanitin to specific cells by macromolecular carriers were 

 patterned after the work of Faulstich and Trischmann (1973) 

 The initial objective was to synthesize a-amanitin-protein 

 conjugates which would then be evaluated on the basis of 

 their biochemical properties and in vitro toxicity for 

 selected cell types. Work by other investigators (Wieland, 

 1968) and NMR studies by Preston and Gabbay (unpublished 

 results, 1977) had demonstrated that a-amanitin contains a 

 hydroxy tryptophan moiety (Figure 1) that is available for 

 chemical cross-linking via diazotization to other aromatic 

 groups. A procedure was developed (Figure 2) from the syn- 

 thesis reported by Faulstich and Trischmann (1973) that 

 allowed for the formation of an a-amanitin derivative, ADH , 

 containing a free amino group. 



Synthesis of compounds leading to the formation of 

 N- (4-aminobenzoyl) -N ' -BOC-hexamethylenediamine was per- 

 formed by Dr. J. F. Preston. This compound was then 

 coupled to the hydroxytryptophan moiety of a-amanitin via 

 a diazo bond and the resulting derivative, ADBH , purified 



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