# US 10,501,762 B2 · Methods and systems for sulfimidation or sulfoximidation of organic molecules

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- Number: US 10,501,762 B2
- Document ID: US10501762B2
- Status: Granted
- Kind: B2
- Application number: 15/208,573
- Filed: 2016-07-12
- Published: 2016-12-08
- Granted: 2019-12-10
- Earliest claimed: 2014-02-18
- Assignees: California Institute of Technology
- Applicants: The California Institute of Technology
- Inventors: Christopher C. Farwell; John A. McIntosh; Frances H. Arnold
- Web page: https://patentgrep.com/patent/US10501762B2

## CPC

- C12P 13/001: {Amines; Imines}
- C12N 9/0042: {NADPH-cytochrome P450 reductase (1.6.2.4)}
- C12P 13/00: Preparation of nitrogen-containing organic compounds
- C12Y 106/02004: NADPH-hemoprotein reductase (1.6.2.4), i.e. NADP-cytochrome P450-reductase
- C12Y 114/14001: Unspecific monooxygenase (1.14.14.1)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES

## Abstract

The disclosure generally relates to the fields of synthetic organic chemistry. In particular, the present disclosure relates to methods and systems for the imidation of sulfides.

## Claims

### Claim 1

A method for catalyzing the intermolecular insertion of nitrogen into organosulfur substrates to produce a product having a new S-N bond, the method comprising:
providing a mixture comprising a nitrene source, an organosulfur substrate and a heme enzyme comprising the amino acid sequence of SEQ ID NO: 1 or an engineered heme enzyme comprising the amino acid sequence of SEQ ID NO: 1 with the exception of at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, or all of the following amino acid substitutions in the amino acid sequence of SEQ ID NO: 1L V78A, F87C, P142S, T175I, A184V, S226R, H236Q, T438S, E252G, A290V, L353V, I366V, C400X, T438S and E442K, wherein “X” is any amino acid other than Cys; and
reacting said mixture for a time sufficient to form a product having a new S-N bond.

### Claim 2 (depends on 1)

The method of claim 1, wherein said nitrene source is an azide.

### Claim 3 (depends on 2)

The method of claim 2, wherein said azide has the general formula R1—N3,
wherein R1 is:
(i) a substituted or unsubstituted aryl, a substituted or unsubstituted aryl, —OR2, or —NR2, wherein R2 is a substituted or unsubstituted aryl, a substituted or unsubstituted alkyl;
(ii) —SO2R3, wherein R3 is a substituted or unsubstituted aryl, a substituted or unsubstituted alkyl, —OR2, or —NR2, wherein R2 are any alkyl or aryl;
(iii) —COR4, wherein R4 is a substituted or unsubstituted aryl, a substituted or unsubstituted alkyl, —OR2, or —NR2, wherein R2 are any alkyl or aryl; or
(iv) —P(O)(OR5)(OR6), wherein R5 and R6 are independently H, a substituted or unsubstituted aryl, a substituted or unsubstituted alkyl.

### Claim 4 (depends on 3)

The method of claim 3, wherein said azide has a structure selected from the group consisting of:
wherein R1 is any alkyl, aryl, —OR, NR2, wherein R, R2 and R3 are any alky, or aryl.

### Claim 5 (depends on 1)

The method of claim 1, wherein said nitrene source is selected from the group consisting of:
wherein R1 is any alkyl, aryl, —OR, NR2, wherein R, R2 and R3 are any alkyl, or aryl.

### Claim 6 (depends on 1)

The method of claim 1, wherein said product having a new S-N bond is an aliphatic amine and said nitrene source is tosyl azide.

### Claim 7 (depends on 1)

The method of claim 1, wherein said product having a new S-N bond is generated through a nitrenoid intermediate.

### Claim 8 (depends on 1)

The method of claim 1, further comprising expressing said heme enzyme in a bacterial, archaeal or fungal host organism.

### Claim 9 (depends on 1)

The method of claim 1, wherein said engineered heme enzyme comprises a T268A mutation, a T438S mutation and/or a C400X mutation in the amino acid sequence of SEQ ID NO: 1, wherein X is any amino acid other than Cys.

### Claim 10 (depends on 1)

The method of claim 1, wherein said product having a new S-N bond is a compound of Formula Ia:
wherein R1 is a sulfoxide, a carbonyl or a phosphonate; wherein R2 is H or any alkyl or aryl; and
wherein R3 is H, O or an optionally substituted aryl group.

### Claim 11 (depends on 10)

The method of claim 10, wherein R1 is a sulfoxide of formula SO2R5, wherein R5 is any alkyl, any aryl, —OR6 or NR7, wherein R6 and Ware any alkyl or any aryl.

### Claim 12 (depends on 10)

The method of claim 10, wherein W is a phosphonate of formula P(O)(OR8)(OR9), wherein R8 and R9 are independently any aryl or any alkyl.