The invention relates to a novel process for the preparation of chiral 2-(4-aminophenyl) morpholines of the formula ##STR00001## wherein R.sup.1 is hydrogen an amino protecting group. The chiral 2-(4-aminophenyl) morpholines of the formula I are key intermediates for the preparation of compounds that have a good affinity to the trace amine associated receptors (TAARs).

The invention relates to a novel process for the preparation of chiral 2-(4-aminophenyl) morpholines of the formula ##STR00001## wherein R.sup.1 is hydrogen an amino protecting group. The chiral 2-(4-aminophenyl) morpholines of the formula I are key intermediates for the preparation of compounds that have a good affinity to the trace amine associated receptors (TAARs).

A method for producing a 1-(acyloxy)alkyl carbamate derivative (III), using a fluorous alkyl carbonate derivative (I), and a fluorous alkyl carbonate derivative (I) and a method for producing the same. In the formula, R.sup.1 represents C.sub.1-C.sub.4 alkyl group or a C.sub.3-C.sub.6 cycloalkyl group, R.sup.2 represents a C.sub.1-C.sub.4 alkyl group or a hydrogen atom, and A represents a fluorous alkyl group (wherein the fluorous alkyl group represents a C.sub.2-C.sub.11 alkyl group in which 40% or more of the hydrogen atoms are replaced by fluorine atoms).

A method for producing a 1-(acyloxy)alkyl carbamate derivative (III), using a fluorous alkyl carbonate derivative (I), and a fluorous alkyl carbonate derivative (I) and a method for producing the same. In the formula, R.sup.1 represents C.sub.1-C.sub.4 alkyl group or a C.sub.3-C.sub.6 cycloalkyl group, R.sup.2 represents a C.sub.1-C.sub.4 alkyl group or a hydrogen atom, and A represents a fluorous alkyl group (wherein the fluorous alkyl group represents a C.sub.2-C.sub.11 alkyl group in which 40% or more of the hydrogen atoms are replaced by fluorine atoms).

The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme including the capability of reducing 5-((4S)-2-oxo-4-phenyl (1,3-oxazolidin-3-yl))-1-(4-fluorophenyl) pentane-1,5-dione to (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize the intermediate (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one in a process for making Ezetimibe.

The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme including the capability of reducing 5-((4S)-2-oxo-4-phenyl (1,3-oxazolidin-3-yl))-1-(4-fluorophenyl) pentane-1,5-dione to (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize the intermediate (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one in a process for making Ezetimibe.

A method for preparing a nitrogen-containing heterocyclic compound and a derivative thereof by an enzymatic-chemical cascade method, comprising: reacting an alcohol, an amine, an alcohol dehydrogenase, a flavin molecule and a coenzyme in a solvent to obtain the nitrogen-containing heterocyclic compound and the derivative thereof; compared with the prior art, the method is a green and economical enzymatic-chemical cascade method, and is used for synthesizing nitrogen-containing heterocyclic compounds and derivatives thereof; compared with a common toxic chemical catalyst, the alcohol dehydrogenase is selected as a catalyst in the method, which has the characteristics of high substrate specificity, no pollution, high catalytic efficiency, no toxic solvents and simple post-treatment.

A method for preparing a nitrogen-containing heterocyclic compound and a derivative thereof by an enzymatic-chemical cascade method, comprising: reacting an alcohol, an amine, an alcohol dehydrogenase, a flavin molecule and a coenzyme in a solvent to obtain the nitrogen-containing heterocyclic compound and the derivative thereof; compared with the prior art, the method is a green and economical enzymatic-chemical cascade method, and is used for synthesizing nitrogen-containing heterocyclic compounds and derivatives thereof; compared with a common toxic chemical catalyst, the alcohol dehydrogenase is selected as a catalyst in the method, which has the characteristics of high substrate specificity, no pollution, high catalytic efficiency, no toxic solvents and simple post-treatment.

The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme including the capability of reducing 5-((4S)-2-oxo-4-phenyl (1,3-oxazolidin-3-yl))-1-(4-fluorophenyl) pentane-1,5-dione to (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize the intermediate (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one in a process for making Ezetimibe.

The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme including the capability of reducing 5-((4S)-2-oxo-4-phenyl (1,3-oxazolidin-3-yl))-1-(4-fluorophenyl) pentane-1,5-dione to (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize the intermediate (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one in a process for making Ezetimibe.