# US 11,078,229 B2 · Thiol compounds and the use thereof for the synthesis of modified oligonucleotides

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 11,078,229 B2
- Document ID: US11078229B2
- Status: Granted
- Kind: B2
- Application number: 15/862,485
- Filed: 2018-01-04
- Published: 2018-09-20
- Granted: 2021-08-03
- Earliest claimed: 2012-04-04
- Assignees: Centre National De La Recherche Scientifique; Etablissement Francais Du Sang
- Applicants: Etablissement Français du Sang; Centre National de la Recherche Scientifique (CNRS)
- Inventors: François Morvan; Albert Meyer; Jean-Jacques Vasseur; Julie Mayen; Carole Chaix; Carole Farre; Chantal Fournier-Wirth; Jean-François Cantaloube; Myriam Lereau
- Web page: https://patentgrep.com/patent/US11078229B2

## CPC

- C07H 21/04: with deoxyribosyl as saccharide radical
- C07C 323/52: the carbon skeleton being acyclic and saturated
- C07F 9/2408: {of hydroxyalkyl compounds}
- C08F 257/02: on to polymers of styrene or alkyl-substituted styrenes
- C12N 15/11: DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity}
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- C07C: ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07F: ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM (metal-containing porphyrins C07D487/22)
- C08F: MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention relates to a thiol compound suitable for forming a chain of oligomers that can be grafted to an oligonucleotide. The invention also relates to an oligonucleotide grafted by such a compound, thus having one or more thiol functions, suitable for being immobilized on a gold surface or on a grafted surface.

## Claims

### Claim 1

A modified oligonucleotide, having the following formula (XIIa):
□—N1—N2— . . . —Nn-1—Nn-(I′)y-M1- . . . -Mm-1-Mm)p-(I′)y′
in which,
N1, . . . Nn represent, independently of one another, a nucleotide,
M1, . . . Mm represent, independently of one another, a nucleotide,
(I′) represents a compound of formula (I′a) or (I′b), where:
where
W is selected from the group consisting of C1-C6 alkane triyl groups, C6-C12 aryl triyl groups, and C6-C12 aralkane triyl groups, wherein the C1-C6 alkane triyl group is a linear or branched C1-C6 alkane triyl substituted by at least two alkyl groups,
Z is selected from the group consisting of C1-C6 alkoxy groups, oxygen-containing or nitrogen-containing C3-C6 cycloheteroalkyl groups, C1-C6 NCO-alkyl groups, and C1-C6 CON-alkyl groups,
Y is selected from the group consisting of linear or branched C1-C6 alkyl groups, C1-C6 aminoalkyl groups, C1-C6 alkoxy groups, C3-C6 cycloalkyl groups, and oxygen-containing or nitrogen-containing C3-C6 cycloheteroalkyl groups,
X is selected from the group consisting of linear or branched C1-C6 alkyl groups, C1-C6 aminoalkyl groups, C1-C6 alkoxy groups, C3-C6 cycloalkyl groups, and oxygen-containing or nitrogen-containing C3-C6 cycloheteroalkyl groups,
R is H, and
R1 is selected from the group consisting of 2-cyanoethyl, R′1R′2R′3SiCH2CH2 groups, and R′1, R′2, R′3, which may be identical or different, and which are selected from the group consisting linear or branched alkyls comprising from 1 to 12 carbon atoms and C6-C12 aryls,
n is an integer comprised between 1 and 100,
m is an integer comprised between 1 and 100,
y is an integer comprised between 1 and 12,
p is 0,
y′ is 0, and
□ represents a solid support.

### Claim 2 (depends on 1)

The modified oligonucleotide according to claim 1, having the formula (XIIc):
in which,
n, y, N1, . . . Nn-1, have the same definition as in claim 1,
X, Y, Z, W have the same definition as in claim 1,
Bn represents the base of the nth nucleotide.

### Claim 3 (depends on 1)

A substrate grafted with at least one modified oligonucleotide according to claim 1, said substrate comprising at least one receiving zone coated with a gold or platinum film or grafted with groups comprising at least one carbon-carbon double bond or a carbon-carbon triple bond or haloacetamide groups, preferably maleimide or acrylamide groups, said substrate being in metal in case of gold film and in plastic in case of grafting with groups comprising at least one carbon-carbon double bond or a carbon-carbon triple bond or haloacetamide groups, preferably maleimide or acrylamide groups.

### Claim 4 (depends on 3)

The substrate according to claim 3, wherein the substrate in metal is in copper or in titanium and/or the substrate in plastic is in polystyrene.

### Claim 5 (depends on 1)

A test kit comprising:
at least one substrate comprising at least one receiving zone coated with a gold or platinum metallic film or a substrate grafted with groups comprising at least one carbon-carbon double bond or a carbon-carbon triple bond or haloacetamide groups, preferably maleimide or acrylamide groups, and
at least one modified oligonucleotide according to claim 1.

### Claim 6 (depends on 3)

A method for using the substrate according to claim 3 comprising using said substrate to produce affinity tests between an oligonucleotide and another molecule.

### Claim 7

A modified oligonucleotide, having the following formula:
(AA)-N1— . . . —Nn-1—Nn-(I′)-(M1- . . . -Mm-1-Mm)p-(I′)b
in which,
N1, . . . Nn represent, independently of one another, a nucleotide,
M1, . . . Mm represent, independently of one another, a nucleotide,
(I′) represents a compound of formula (I′a) or (I′b), where:
wherein
W is selected from the group consisting of C1-C6 alkane triyl groups, C6-C12 aryl triyl groups, and C6-C12 aralkane triyl groups, wherein the C1-C6 alkane triyl group is a linear or branched C1-C6 alkane triyl substituted by at least two alkyl groups,
Z is selected from the group consisting of C1-C6 alkoxy groups, oxygen-containing or nitrogen-containing C3-C6 cycloheteroalkyl groups, C1-C6 NCO-alkyl groups, and C1-C6 CON-alkyl groups,
Y is selected from the group consisting of linear or branched C1-C6 alkyl groups, C1-C6 aminoalkyl groups, C1-C6 alkoxy groups, C3-C6 cycloalkyl groups, and oxygen-containing or nitrogen-containing C3-C6 cycloheteroalkyl groups,
X is selected from the group consisting of linear or branched C1-C6 alkyl groups, C1-C6 aminoalkyl groups, C1-C6 alkoxy groups, C3-C6 cycloalkyl groups, and oxygen-containing or nitrogen-containing C3-C6 cycloheteroalkyl groups,
R is H, and
R1 is selected from the group consisting of 2-cyanoethyl, R′1R′2R′3SiCH2CH2 groups, and R′1, R′2, R′3, which may be identical or different, and which are selected from the group consisting of linear or branched alkyls comprising from 1 to 12 carbon atoms, and C6-C12 aryls,
n is an integer comprised between 1 and 100,
m is an integer comprised between 1 and 100,
y is an integer comprised between 1 and 12,
(AA) is a solid support or (Ic)-(I′)y-1,
wherein when (AA) is a solid support,
a=y and b=y′, p is 0, and y′ is 0, and
wherein when (AA) is (Ic)-(I′)y-1,
a=y′ and b=y″, and y′ is 0, and y″ is 0,
wherein (Ic) is
and
wherein W, Z, X, Y, and R are defined above, D is a protective group of the alcohols that allow orthogonal deprotection with respect to the other groups of compound (I′a), (I′b), or (Ic), J is selected from the group consisting of a single bond and a —CH2—, —CH2CH2—, —CH2OCH2—, or —CH2OPhOCH2— group where Ph is a benzyl, and □ represents a solid support.