Thiol compounds and the use thereof for the synthesis of modified oligonucleotides

Patent metadata

Field	Value
Publication number	US-11078229-B2
Application number	US-201815862485-A
Country	US
Kind code	B2
Filing date	Jan 4, 2018
Priority date	Apr 4, 2012
Publication date	Aug 3, 2021
Grant date	Aug 3, 2021

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.

First claim

What is claimed is:

1. A modified oligonucleotide, having the following formula (XIIa):
□—N 1 —N 2 — . . . —N n-1 —N n -(I′) y -M 1 - . . . -M m-1 -M m ) p -(I′) y′

in which,
N 1 , . . . N n represent, independently of one another, a nucleotide,
M 1 , . . . M m 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
R 1 is selected from the group consisting of 2-cyanoethyl, R′ 1 R′ 2 R′ 3 SiCH 2 CH 2 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.

2. The modified oligonucleotide according to claim 1 , having the formula (XIIc):

in which,
n, y, N 1 , . . . N n-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 n th nucleotide.

3. 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.

4. 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.

5. 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 .

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

7. A modified oligonucleotide, having the following formula:
(AA)-N 1 — . . . —N n-1 —N n -(I′)-(M 1 - . . . -M m-1 -M m ) p -(I′) b

in which,
N 1 , . . . N n represent, independently of one another, a nucleotide,
M 1 , . . . M m 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
R 1 is selected from the group consisting of 2-cyanoethyl, R′ 1 R′ 2 R′ 3 SiCH 2 CH 2 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 —CH 2 —, —CH 2 CH 2 —, —CH 2 OCH 2 —, or —CH 2 OPhOCH 2 — group where Ph is a benzyl, and □ represents a solid support.

Assignees

• Francais Du Sang Ets
• Centre Nat Rech Scient

Inventors

• Morvan François
• Meyer Albert
• Vasseur Jean-Jacques
• Mayen Julie
• Chaix Carole
• Farre Carole
• Fournier-Wirth Chantal
• Cantaloube Jean-François
• Lereau Myriam

Classifications

• C07H21/04 Primary

  with deoxyribosyl as saccharide radical · CPC title

• C07F9/2408

  of hydroxyalkyl compounds · CPC title

• C07C323/52

  the carbon skeleton being acyclic and saturated · CPC title

• C12N15/11

  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} · CPC title

• C08F257/02

  on to polymers of styrene or alkyl-substituted styrenes · CPC title