In vivo and in vitro olefin cyclopropanation catalyzed by heme enzymes

摘要

The present invention provides methods for catalyzing the conversion of an olefin to any compound containing one or more cyclopropane functional groups using heme enzymes. In certain aspects, the present invention provides a method for producing a cyclopropanation product comprising providing an olefinic substrate, a diazo reagent, and a heme enzyme; and admixing the components in a reaction for a time sufficient to produce a cyclopropanation product. In other aspects, the present invention provides heme enzymes including variants and fragments thereof that are capable of carrying out in vivo and in vitro olefin cyclopropanation reactions. Expression vectors and host cells expressing the heme enzymes are also provided by the present invention.

主权项

1. A reaction mixture for producing a cyclopropanation product, the reaction mixture comprising an olefinic substrate, a carbene precursor, and a heme enzyme, wherein the cyclopropanation product is a compound according to Formula I: wherein:
R1 is independently selected from the group consisting of H, optionally substituted C1-18 alkyl, optionally substituted C6-10 aryl, optionally substituted 5- to 10-membered heteroaryl, C(O)OR1a, and C(O)R8;R2 is independently selected from the group consisting of H, optionally substituted C1-18 alkyl, optionally substituted C6-10 aryl, optionally substituted 5- to 10-membered heteroaryl, C(O)OR2a, and C(O)R8; andR3, R4, R5, and R6 are independently selected from the group consisting of H, C1-18 alkyl, optionally substituted C6-10 aryl, optionally substituted C1-C6 alkoxy, NR7C(O)R8, C(O)R8, C(O)OR8, and N(R9)2; wherein:
R1a and R2a are independently selected from the group consisting of H and optionally substituted C1-18 alkyl;each R7 and R8 is independently selected from the group consisting of H, optionally substituted C1-12 alkyl, and optionally substituted C6-10 aryl; andtwo R9 moieties, together with the nitrogen atom to which they are attached, form an optionally substituted 5- to 10-membered heterocyclyl.