# US 2019/0169228 A1 · Selective disulfidation reagent using nitrogen-containing compound and method for producing disulfide-containing compound

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- Number: US 2019/0169228 A1
- Document ID: US20190169228A1
- Status: Published application
- Kind: A1
- Application number: 16/303,008
- Filed: 2017-05-22
- Published: 2019-06-06
- Earliest claimed: 2016-05-20
- Assignees: Tokyo University of Pharmacy & Life Sciences; Kokusan Chemical Co., Ltd.
- Applicants: Tokyo University of Pharmacy & Life Sciences; Kokusan Chemical Co., Ltd.
- Inventors: Yoshio Hayashi; Akihiro Taguchi; Kentaro Fukumoto
- Web page: https://patentgrep.com/patent/US20190169228A1

## CPC

- C07D 401/14: containing three or more hetero rings
- C07K 1/086: {containing sulfur}
- C07D 213/80: in position 3
- C07D 401/04: directly linked by a ring-member-to-ring-member bond
- C07D 401/12: linked by a chain containing hetero atoms as chain links
- C07D 417/04: directly linked by a ring-member-to-ring-member bond
- C07D 417/14: containing three or more hetero rings
- C07K 1/02: in solution {(C07K1/003, C07K1/006 take precedence)}
- C07D 471/04: Ortho-condensed systems
- C07K 1/061: {using protecting groups}
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

The present invention provides a means capable of selectively introducing a disulfide bond with respect to two free thiol groups located in a molecule of an organic compound such as a peptide, or the like, in a short time by a simple treatment and also by a chemically stable method. A nitrogen-containing compound represented by Chemical Formula 1 below or a salt thereof: The symbols shown in Chemical Formula 1 are the same as defined in the specification.

## Claims

### Claim 1

A nitrogen-containing compound represented by Chemical Formula 1 below or a salt thereof:
wherein
W, together with other ring member atoms, forms a nitrogen-containing heterocycle selected from the group consisting of a pyridine ring, a pyrazine ring, an imidazole ring, an oxazole ring, a thiazole ring, a quinoline ring, an isoquinoline ring, a quinoxaline ring, a phenanthroline ring, a pteridine ring, and an azocine ring,
X is —O— or —NH—,
Y is selected from the group consisting of a substituted or unsubstituted C1-C20 alkyl group, a substituted or unsubstituted C2-C20 alkenyl group, a substituted or unsubstituted C2-C20 alkynyl group, a substituted or unsubstituted C3-C20 cycloalkyl group, a substituted or unsubstituted C3-C20 cycloalkenyl group, a substituted or unsubstituted C6-C20 aryl group, a substituted or unsubstituted C3-C20 heteroaryl group, and a monovalent group derived from a substituted or unsubstituted aliphatic heterocycle having an electron-withdrawing property,
Z represents a hydrogen atom or an electron-withdrawing substituent present on the nitrogen-containing heterocycle,
p, q, and r are each independently 0 or 1,
s represents an integer of 0 to 10,
L0 and L1 each independently represent a linker having a chemically stable structure,
Aa and Ab are each independently a group selected from the group consisting of —CH═CH—, —C≡C—, —C(═O)—, —C(═O)—O—, —O—C(═O)—, —O—, C1-C20 oxyalkylene group, C1-C20 alkyleneoxy group, —C(═O)—NH—, —NH—C(═O)—, —NH—C(═O)—NH—, hydrazine, triazole, sulfone, sulfoxide, sulfonic acid ester, sulfonamide, sulfinic acid ester, sulfinamide, piperidine, and dioxane, and
R is a hydrogen atom, a substituted or unsubstituted C1-C20 alkyl group, a substituted or unsubstituted C2-C20 alkenyl group, a substituted or unsubstituted C2-C20 alkynyl group, a substituted or unsubstituted C3-C20 cycloalkyl group, a substituted or unsubstituted C3-C20 cycloalkenyl group, a substituted or unsubstituted C6-C20 aryl group, a substituted or unsubstituted C3-C20 heteroaryl group, an amino group, a hydroxy group, or a polymer carrier;
provided that the following compounds (A) to (H) are excluded:
(A) 3-nitro-2-pyridine sulfinic acid methyl,
(B) 3-nitro-2-pyridine sulfinic acid ethyl,
(C) 3-nitro-2-pyridine sulfinic acid N,N-diethylaminoethyl,
(D) N-(3′-nitro-T-pyridinesulfenyloxy)-5-norbornene-2,3-dicarboxyimide,
(E) (S)-((tert-butoxycarbonyl)amino)-3-(((3-nitropyridin-2-yl)thio)oxy)propanoic acid,
(F) (2S,3R)-2-(((benzyloxy)carbonyl)amino)-3-(((3-nitropyridin-2-yl)thio)oxy)butanoic acid,
(G) 4-((((3-nitropyridin-2-yl)thio)oxy)methyl)benzoic acid, and
(H) (S)-2-(((3-nitropyridin-2-yl)thio)oxy)-3-phenylpropanoic acid.

### Claim 2 (depends on 1)

The nitrogen-containing compound or a salt thereof according to claim 1, wherein the nitrogen-containing compound or the salt thereof does not satisfy the following conditions: Conditions: p=0, s=0, r=0, and R=hydrogen atom.

### Claim 3 (depends on 1)

The nitrogen-containing compound or a salt thereof according to claim 1, wherein W, together with other ring member atoms, forms a pyridine ring as the nitrogen-containing heterocycle.

### Claim 4 (depends on 1)

The nitrogen-containing compound or a salt thereof according to claim 1, wherein X is —O—.

### Claim 5 (depends on 1)

The nitrogen-containing compound or a salt thereof according to claim 1, wherein Y is a substituted or unsubstituted C1-C20 alkyl group, or a substituted or unsubstituted C6-C20 aryl group.

### Claim 6 (depends on 5)

The nitrogen-containing compound or a salt thereof according to claim 5, wherein Y is a substituted or unsubstituted C1-C20 alkyl group.

### Claim 7 (depends on 1)

The nitrogen-containing compound or a salt thereof according to claim 1, wherein Z is an electron-withdrawing substituent.

### Claim 8 (depends on 7)

The nitrogen-containing compound or a salt thereof according to claim 7, wherein the electron-withdrawing substituent is a nitro group, a trifluoromethyl group, or a halogen atom.

### Claim 9 (depends on 1)

The nitrogen-containing compound or a salt thereof according to claim 1, wherein L0 and L1 are each independently a substituted or unsubstituted C1-C20 alkylene group, a substituted or unsubstituted C2-C20 alkenylene group, a substituted or unsubstituted C2-C20 alkynylene group, a substituted or unsubstituted C3-C20 cycloalkylene group, a substituted or unsubstituted C3-C20 cycloalkenylene group, a substituted or unsubstituted C6-C20 arylene group, a substituted or unsubstituted C3-C20 heteroarylene group, —NH—, —O—, —S—, —C(═O)—NH—, —NH—C(═O)—, —O—, —C(═O)—O—, —O—C(═O)—, —S—, —C(═O)—, a polyoxyalkylene group, and a group represented by Chemical Formula (a) below:
wherein Ra represents and is selected from the group consisting of a substituted or unsubstituted C1-C15 alkylene group wherein these alkylene groups, alkenylene groups, alkynylene groups, cycloalkylene groups, cycloalkenylene groups, arylene groups, and heteroarylene groups may have a substituent.

### Claim 10 (depends on 1)

The nitrogen-containing compound or a salt thereof according to claim 1, wherein the nitrogen-containing compound is represented by Chemical Formula 2 below:
wherein
W, X, Y, Z, p, L0, and Aa are the same as defined in claim 1, and
R is a hydrogen atom, a substituted or unsubstituted C1-C20 alkyl group, a substituted or unsubstituted C2-C20 alkenyl group, a substituted or unsubstituted C2-C20 alkynyl group, a substituted or unsubstituted C3-C20 cycloalkyl group, a substituted or unsubstituted C3-C20 cycloalkenyl group, a substituted or unsubstituted C6-C20 aryl group, or a substituted or unsubstituted C3-C20 hetero aryl group.

### Claim 11 (depends on 10)

The nitrogen-containing compound or a salt thereof according to claim 10, wherein R is a substituted or unsubstituted C1-C20 alkyl group.

### Claim 12 (depends on 10)

The nitrogen-containing compound or a salt thereof according to claim 10, wherein Aa is selected from the group consisting of —C(═O)—, —C(═O)—O—, —O—C(═O)—, —O—, —C(═O)—NH—, and —NH—C(═O)—.

### Claim 13 (depends on 12)

The nitrogen-containing compound or a salt thereof according to claim 12, wherein Aa is —C(═O)—O—.

### Claim 14 (depends on 10)

The nitrogen-containing compound or a salt thereof according to claim 10, wherein p is 0.

### Claim 15 (depends on 1)

The nitrogen-containing compound or a salt thereof according to claim 1, wherein R is a polymer carrier.

### Claim 16 (depends on 15)

The nitrogen-containing compound or a salt thereof according to claim 15, wherein R is a polymer carrier used in a solid phase synthesis method.

### Claim 17 (depends on 15)

The nitrogen-containing compound or a salt thereof according to claim 15, wherein R is selected from the group consisting of polystyrene, polypropylene, polyethylene, polyether, polyvinyl chloride, dextran, polyacrylamide, polyethylene glycol, copolymers and crosslinked products thereof, magnetic beads, and a combination thereof.

### Claim 18 (depends on 1, 15)

The nitrogen-containing compound or a salt thereof according to claim 15, wherein the nitrogen-containing compound is represented by Chemical Formula 3 below:
wherein W, X, Y, Z, p, L0, and Aa are the same as defined in claim 1, and R is a polymer carrier.

### Claim 19 (depends on 18)

The nitrogen-containing compound or a salt thereof according to claim 18, wherein Aa is selected from the group consisting of —C(═O)—, —C(═O)—O—, —O—C(═O)—, —O—, —C(═O)—NH—, and —NH—C(═O)—.

### Claim 20 (depends on 19)

The nitrogen-containing compound or a salt thereof according to claim 19, wherein Aa is —C(═O)—NH—, and R is a polyethylene glycol crosslinked product.

### Claim 21 (depends on 19)

The nitrogen-containing compound or a salt thereof according to claim 19, wherein Aa is —C(═O)—O—, and R is a polystyrene resin.

### Claim 22 (depends on 15)

The nitrogen-containing compound or a salt thereof according to claim 15, wherein p is 0.

### Claim 23 (depends on 1, 15)

The nitrogen-containing compound or a salt thereof according to claim 15, wherein the nitrogen-containing compound is represented by Chemical Formula 4 below:
wherein W, X, Y, Z, p, L0, L1, Aa, and Ab are the same as defined in claim 1, and R is a polymer carrier.

### Claim 24 (depends on 23)

The nitrogen-containing compound or a salt thereof according to claim 23, wherein Aa and Ab are —C(═O)—NH—, L1 is a C1-C20 alkylene group, and R is a polyethylene glycol crosslinked product.

### Claim 25 (depends on 23)

The nitrogen-containing compound or a salt thereof according to claim 23, wherein p is 0.

### Claim 26 (depends on 1)

A disulfidation reagent for a thiol group, comprising:
the nitrogen-containing compound or a salt thereof according to claim 1, or any one of the following compounds (A) to (H):
(A) 3-nitro-2-pyridine sulfinic acid methyl,
(B) 3-nitro-2-pyridine sulfinic acid ethyl,
(C) 3-nitro-2-pyridine sulfinic acid N,N-diethylaminoethyl,
(D) N-(3′-nitro-2′-pyridinesulfenyloxy)-5-norbornene-2,3-dicarboxyimide,
(E) (S)-((tert-butoxycarbonyl)amino)-3-(((3-nitropyridin-2-yl)thio)oxy)propanoic acid,
(F) (2S,3R)-2-(((benzyloxy)carbonyl)amino)-3-(((3-nitropyridin-2-yl)thio)oxy)butanoic acid,
(G) 4-((((3-nitropyridin-2-yl)thio)oxy)methyl)benzoic acid, and
(H) (S)-2-(((3-nitropyridin-2-yl)thio)oxy)-3-phenylpropanoic acid.

### Claim 27 (depends on 26)

The disulfidation reagent according to claim 26, wherein R in the nitrogen-containing compound is a polymer carrier used in a solid phase synthesis method.

### Claim 28 (depends on 26)

A method for producing a disulfide-containing compound, the method comprising:
contacting a compound having two or more free thiol groups in a molecule with the disulfidation reagent according to claim 26 to form a disulfide bond between the two free thiol groups, thereby obtaining a disulfide-containing compound.

### Claim 29 (depends on 28)

The method according to claim 28, wherein the disulfide-containing compound is a disulfide-containing peptide.

### Claim 30 (depends on 1)

A method for producing the nitrogen-containing compound or a salt thereof according to claim 1, the method comprising:
(I) reacting a compound represented by Chemical Formula 5 below with a halogen simple substance or a halogen generating reagent to produce a compound represented by Chemical Formula 6 below;
wherein W, Z, p, q, r, s, L0, L1, Aa, and Ab are the same as defined in claim 1, and R″ is a leaving group,
wherein W, Z, p, q, r, s, L0, L1, Aa, and Ab are the same as defined in claim 1, and Hal represents a halogen atom selected from fluorine, chlorine, bromine or iodine,
and
(II) reacting the compound represented by Chemical Formula 6 with an alcohol represented by Y—OH or an amine represented by Y—NH2 under basic conditions, wherein Y is the same as defined in Chemical Formula 1, thereby producing a nitrogen-containing compound represented by Chemical Formula 1 below:
wherein W, X, Y, Z, p, q, r, s, L0, L1, Aa, and Ab are the same as defined in claim 1.