Statin side chain

Bergeron, S., Chaplin, D.A., Edwards, J.H. et al. (2006) Nitrilase-catalyzed desymmetrization of 3-hydroxy-glutaronitrile preparation of a statin side-chain intermediate. Organic Process Research Development, 10, 661-665. 

Bergeron S, Chaplin D, Edwards JH, Ellis BS, Hill CL, Holt-Tiffin K, Knight JR, Mahoney T, Osborne AP, Ruecroft G (2006) Nitrilase-catalyzed desym-metrization of 3-hydroxyglutaronitrile preparation of a statin side-chain intermediate. Org Proc Res Dev 10 661-665 Burns M, Weaver J, Wong J (2005) Stereoselective enzymic bioconversion of aliphatic dinitriles into cyano carboxylic acids. WO 2005100580 DeSantis G, Zhu Z, Greenberg W, Wong K, Chaplin J, Hanson SR, Farwell B, Nicholson LW, Rand CL, Weiner DP, Robertson D, Burk MJ (2002) An enzyme library approach to biocatalysis development of nitrilases for enantioselective production of carboxylic acid derivatives. J Am Chem Soc 124 9024-9025... 

Scheme 1.50 Nitrilase desymmetrization approach to the atorvastatin statin side chain...

Several approaches to statin side-chain intermediates have so far been discussed. Whereas these chemoenzymatic approaches provide clear benefits over the chemical processes, they do not harness the tme potential of biocatalysis as the biotransformations have simply been inserted into the existing chemical route. Wong and co-workers have developed a more biosynthetic-hke approach by using a mutant 2-deoxyiibose-5-phosphate aldolase (DERA)... 

Of the known classes of aldolase, DERA (statin side chain) and pyruvate aldolases (sialic acids) have been shown to be of particular value in API production as they use readily accessible substrates. Glycine-dependent aldolases are another valuable class that allow access to p-hydroxy amino acid derivatives. In contrast, dihydroxy acetone phosphate (DHAP) aldolases, which also access two stereogenic centres simultaneously,... 

Muller, M., Chemoenzymatic s3mthesis of building blocks for statin side chains. Angew. Chem. Int. Ed., 2005, 44, 362-365. 

Sustainable Chemoenzymatic Processes for Statin Side Chains... 

A classical chemical synthesis of a statin side chain is shown in Scheme 6.1. Typically, a P-keto ester is reduced to set the first chiral center, and the Claisen... 

Scheme 6.1 Typical chemical route to statin side chains.

Biocatalysis plays a central role in the manufacturing of statin side chains (Figure 6.2). A first set of approaches exploits enzymatic desymmetrization reactions, for example, of the methoxyacetyl ester of glutaric acid diethyl ester with commercially available a-chymotrypsin as explored by Ciba SC with a yield of 94% and enantiomeric excess of up to 98% [1]. In the optimized procedure, the substrate was available in a concentration of 1 M at an enzyme/substrate ratio of 7% (wt/wt), and the reaction took approximately a day. The subsequent steps to the final acetonide also involved a pig-liver esterase (PLE) catalyzed selective hydrolysis of the methoxyacetyl group (Figure 6.2a). 

The beauty and the economic advantage of the DERA-based processes to certain statin side chains is based on the fact that the carbon skeleton and the two chiral centers of the side chains are built up in one step from the simple, cheap starting materials acetaldehyde and chloroacetaldehyde. This compensates for the higher biocatalyst loading required by the DERA approach compared with the other bio-catalytic processes summarized above, as will be described in the following sections. 

To overcome problems of poor acceptor substrate acceptance, high concentrations of aldehyde substrates are required to obtain synthetically useful product yields. Unfortunately, DERA shows rather poor resistance to such high aldehyde concentrations, especially toward CIAA, resulting in rapid, irreversible inactivation of the enzyme. Therefore, the organic synthesis of (3R,5S)-6-chloro-2,4,6-trideoxy-hexapyranoside 1 requires very high amounts of DERA. Thus, despite the synthetic usefulness of DERA to produce chiral intermediates for statin side chains, the large-scale application is seriously hampered by its poor stability at industrially relevant aldehyde concentrations. The production capacity for such 2,4,6-trideoxy-hexoses of wild-type E. coli DERA is rather low [15]. 

Several efficient biocatalytic processes to statin side chain intermediates have been developed in the last two decades, and all have their characteristic advantages and... 

This, together with the short synthetic route starting from simple and cheap raw materials and the applicability of the DERA product as a common statin building block, makes the DERA process one of the economically most attractive routes for the manufacture of statin side chain intermediates. 

Another way to statin side chains, via the intermediate syn-(i K,5,S )-6-chloro-hexanoate, employs regioselective and (K)-specific reduction with alcohol dehydrogenase (ADH) from Lactobacillus brevis to yield the intermediate (5S)-6-chloro-3-ketohexanoate from the 3,5-diketo acid (Wolberg, 2001) (Figure 13.16). Further reduction of (5S)-6-chloro-3-ketohexanoate to syn-(3R,5S)-6-chlorohexanoate is afforded chemically with NaBH4/B(OMe)Et2. 

In another development, the statin side chain en route to Atorvastatin (Lipitor , Pfizer) is synthesized via the key intermediate alkyl 3-hydroxy-4-cyanobutyrate (Figure 13.17). Instead of the currently practiced six-step route, a much more concise three-step route starts from epichlorohydrin via Cl chain length enhancement by both nucleophilic substitution of chloride and nucleophilic ring opening of the epoxide with cyanide to yield symmetric dicyanoisopropanol. Nitrilase action desymmetrizes the dinitrile intermediate with the creation of a chiral center in C3 to yield (R)-3-hydroxy-4-cyanobutyrate, which is esterified to the key intermediate ethyl (R)-3-hydroxy-4-cyanobutyrate. 

Fig. 17 Ketoreductase-catalyzed chemoenzymatic synthesis of statin side chains...

Scheme 6.19 Enantioselective CIBA route towards the statin side chains, based on a symmetric starting material.

ADH from Lactobacillus brevis (IBADH) was used for the synthesis of a statin side chain, used as an alternative key intermediate in the synthesis of atorvastatin (Figure 13.2). In this variant, the NADPH-dependent enzyme was highly regio-and stereoselective, reducing fert-butyl 6-chloro-3,5-dioxohexanoate to tert-butyl (S)-6-chloro-5-hydroxy-3-oxohexanoate in 72% yield and with >99.5% ee. The cofactor was recycled in a coupled-substrate approach [1], where isopropanol was concomitantly oxidized to acetone at the expense of NADP, thereby driving the reaction toward product formation. Crude cell extract of recombinant LBADH expressed in E. coli was used in a fed-batch system on 8 1 scale, and a TTN of 2x10 could be calculated for IBADH [13-15]. 

S. Bergeron, D.A. Chaplin, J.H. Edwards, B.S.W. Ellis, C.L. Hill, K. Holt-Tiffin, J.R. Knight, T. Mahoney, A.P. Osborne, G. Ruecroft, Nitrilase-catalysed desymmetrisation of 3-hydroxyglutaronitrile preparation of a statin side-chain intermediate, Org. Process Res. Dev. 10 (2006) 661-665. 

---