Amatoxins are ribosomally synthesized and post-translationally modified bicyclic octapeptides biosynthesized by the deadly basidiomycete fungus Amanita phalloides. Amongst this group, alpha-amanitin is the most widely known toxin and is currently under investigation as part of an antibody-drug conjugate to treat pancreatic cancer. As a consequence, an efficient, modular synthetic route to the molecule is highly desirable.
This thesis describes our efforts toward the development of a novel synthesis of alpha-amanitin. An unprecedented tryptathionine formation was envisioned as the key step, which would generate the characteristic cross-linker between cysteine and tryptophan and allow for late stage diversification of the molecule. The transformation would be realized in analogy to a report by Fagnou and co-workers, describing a Rh(III)-catalyzed oxidative annulation of aniline derivatives with internal alkynes to form 2,3-disubstituted indoles.

In this effort, several methods for the regioselective formation of 2-substituted indoles were identified in studies with a model thioalkyne. 3,4-Dimethoxybenzyl was chosen as an orthogonal protecting group for the indole nitrogen, and the regioselectivity with respect to substitution on the ring was controlled by exploiting the sterics of the substituent. A method for the selective preparation of 3 sulfenyl indoles was also identified.

For several decades, the total synthesis of amanitin had been prevented due to the lack of an efficient method to prepare the characteristic unnatural amino acid dihydroxyisoleucine (DhIle). The present thesis describes a novel, efficient and modular synthesis of this challenging scaffold. The crucial anti-diastereselectivity in the generation of its characteristic stereocenters was obtained in an organocatalyzed asymmetric Mannich reaction/olefination protocol. To avoid lactonization, an alternative synthetic route to a DhIle-containing dipeptide 234 was developed, allowing for the efficient preparation of the valuable diol on a larger scale.

Subsequently, preliminary results demonstrated the feasibility of a proposed Pd(II)-catalyzed thioalkynylation by the synthesis of a model dipeptide, and its applicability on a model macrocyclic peptide scaffold was shown. This was later used as a proof of concept for the key Cp*Rh(III)-catalyzed tryptathionine formation. Unfortunately, the isolation of this bicyclic octapeptide was so far unsuccessful, as was the preparation of the target amanitin scaffold. As a consequence, an alternative synthetic strategy to this peptide scaffold was developed. Future works would include its preparation on a larger scale to evaluate the key transformation and access the target alpha-amanitin.

The envisioned strategy for the synthesis of alpha-amanitin was also probed for the potential preparation of phalloidin, another potent compound produced by the Death Cap mushroom, with interesting applications as a biological tool. In this effort, a novel synthetic route to a dipeptide containing the characteristic dihydroxyleucine moiety was developed. A route to the cyclic phalloidin scaffold was explored, as well as a potential alternative route to the linear scaffold to allow for efficient scaling of the synthesis.