Inorganic porous carrier, and production method for nucleic acid using same

Patent metadata

Field	Value
Publication number	US-12377409-B2
Application number	US-202017599700-A
Country	US
Kind code	B2
Filing date	Feb 28, 2020
Priority date	Mar 29, 2019
Publication date	Aug 5, 2025
Grant date	Aug 5, 2025

Abstract

An inorganic porous carrier including a linker of formula (1) and having mode diameter of 0.04 μm to 1 μm in a pore distribution and the density of voids having an opening area of 0.0025 μm 2 or more of 12 to 30 voids/μm 2 [a bond * represents a linkage to the oxygen atom in a silanol group of an inorganic porous substance; n is an integer; R represents an alkyl group containing 3 to 10 carbon atoms which may optionally have a substituent such as an alkoxy group; and L represents a single bond; an alkylene group of 1 to 20 carbon atoms; or an alkylene group containing 2 to 20 carbon atoms containing —CH 2 -Q-CH 2 — group wherein Q selected from a group consisting of —O— etc. is inserted into at least one of —CH 2 —CH 2 — group constituting the alkylene group]; and a method for preparing a nucleic acid using the same.

First claim

The invention claimed is:

1. An inorganic porous carrier, comprising:
a linker of formula (1); and
an inorganic porous substance connected to the linker,
wherein the inorganic porous substance has a most frequent value of pore diameter, which is a mode diameter, ranging from 0.04 μm to 1 μm in a pore distribution as measured by mercury intrusion method, and the inorganic porous substance has a density of voids from 12 voids/μm 2 to 30 voids/μm 2 , the voids having an opening area of 0.0025 m 2 or more as obtained by image analysis of a surface of the inorganic porous substance,

wherein,
a bond marked with * is a linkage to an oxygen atom of a silanol group in the inorganic porous substance;
n is an integer of 1, 2 or 3;
R is independently of each other an alkyl group including 3 to 10 carbon atoms which may optionally have a substituent selected from the group consisting of an alkoxy group and a fluorine atom; a phenyl group which may optionally have a substituent selected from the group consisting of an alkyl group, an alkoxy group, and a fluorine atom; a hydroxyl group; or an alkoxy group including 1 to 4 carbon atom; and
L is a single bond; an alkylene group of 1 to 20 carbon atoms; or an alkylene group including 2 to 20 carbon atoms which includes —CH 2 -Q-CH 2 — group wherein Q is selected from the group consisting of —O—, —NH—, —NH—CO— and —NH—CO—NH—, Q is inserted into at least one of —CH 2 —CH 2 — group constituting the alkylene group, and a carbon atom of a methylene group bonded to Q is not bonded to another group Q.

2. An inorganic porous carrier, comprising:
a linker of formula (2); and
an inorganic porous substance connected to the linker,
wherein the inorganic porous substance has a most frequent value of Pore diameter, which is a mode diameter, ranging from 0.04 μm to 1 pm in a pore distribution as measured by mercury intrusion method, and the inorganic porous substance has a density of voids from 12 voids/μm 2 to 30 voids/μm 2 , the voids having an opening area of 0.0025 μm 2 or more as obtained by image analysis of a surface of the inorganic porous substance,

wherein,
a bond marked with * is a linkage to an oxygen atom of a silanol group in the inorganic porous substance;
n is an integer of 1, 2 or 3;
R is independently of each other an alkyl group including 3 to 10 carbon atoms which may optionally have a substituent selected from the group consisting of an alkoxy group and a fluorine atom; a phenyl group which may optionally have a substituent selected from the group consisting of an alkyl group, an alkoxy group, and a fluorine atom; a hydroxyl group; or an alkoxy group including 1 to 4 carbon atom;
L is a single bond; an alkylene group including 1 to 20 carbon atoms; or an alkylene group including 2 to 20 carbon atoms which includes —CH 2 -Q-CH 2 — group wherein Q is selected from the group consisting of —O—, —NH—, —NH—CO—, and —NH—CO—NH—, Q is inserted into at least one —CH 2 —CH 2 — group constituting the alkylene group, and a carbon atom of a methylene group bonded to Q is not bonded to another group Q;
R b is a nucleoside or a nucleotide; and
L 1 is a divalent group bonded to an oxygen atom of a primary or a secondary hydroxy group of R b .

3. The inorganic porous carrier according to claim 1 , wherein the inorganic porous substance has a specific surface area per volume of 0.1 m 2 /mL to 100 m 2 /mL.

4. The inorganic porous carrier according to claim 1 , wherein the inorganic porous substance has a pore volume per volume of 0.05 mL/mL to 0.6 mL/mL.

5. The inorganic porous carrier according to claim 1 , wherein the inorganic porous substance has a porosity of 50% or more.

6. The inorganic porous carrier according to claim 2 , wherein the linker has a density of 0.1 μmol/m 2 -5.0 μmol/m 2 relative to a specific surface area per mass of the inorganic porous substance.

7. The inorganic porous carrier according to claim 1 , wherein the inorganic porous substance has a median diameter of 1 μm to 1000 μm.

8. The inorganic porous carrier according to claim 1 , wherein the inorganic porous substance comprises at least one selected from the group consisting of silica, silica gel, zeolite, and glass.

9. The inorganic porous carrier according to claim 2 , wherein L 1 includes a succinyl group as a functional group.

10. A method for preparing a nucleic acid with the inorganic porous carrier of the formula (2) of claim 2 , wherein R b is a nucleoside or a nucleotide, which includes a hydroxyl group as a reactive group that is protected, the method comprising:
deprotecting a protecting group of the hydroxyl group at a 5′ position of the nucleoside of R b ;
subjecting the hydroxyl group at the 5′ position to a condensation reaction with an amidite compound having a nucleoside base to produce a phosphite;
oxidizing the phosphite to produce a nucleotide including a protected hydroxyl group at a 5′ position thereof; and
deprotecting a protecting group of the protected hydroxyl group at the 5′ position of the nucleotide to obtain a deprotected product.

11. The method according to claim 10 , further comprising:
(B′) subjecting the deprotected product to a condensation reaction with an amidite compound having a nucleoside base to produce a phosphite;
(C′) oxidizing the phosphite produced in (B′) to produce an oligonucleotide including a protected hydroxyl group at a 5′ position at an end thereof; and
(D′) deprotecting a protecting group of the protected hydroxyl group at the 5′ position at the end of the oligonucleotide to obtain a deprotected product.

12. The method according to claim 11 , further comprising:
repeating (B′), (C′) and (D′) to produce an elongated nucleic acid; and
cleaving the elongated nucleic acid from the inorganic porous carrier.

13. The inorganic porous carrier according to claim 2 , wherein the inorganic porous substance has a specific surface area per volume of 0.1 m 2 /mL to 100 m 2 /mL.

14. The inorganic porous carrier according to claim 2 , wherein the inorganic porous substance has a pore volume per volume of 0.05 mL/mL to 0.6 mL/mL.

15. The inorganic porous carrier according to claim 2 , wherein the inorganic porous substance has a porosity of 50% or more.

16. The inorganic porous carrier according to claim 2 , wherein the inorganic porous substance has a median diameter of 1 μm to 1000 μm.

17. The inorganic porous carrier according to claim 2 , wherein the inorganic porous substance comprises at least one selected from the group consisting of silica, silica gel, zeolite, and glass.

18. The method according to claim 12 , wherein the cleaving of the elongated nucleic acid is performed by ammonia or an amine.

Assignees

• Sumitomo Chemical Co

Inventors

• Arimura Takashi
• Kitahara Masaki
• Yoshioka Takeshi
• Hara Takashi

Classifications

• C07H21/00

  Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids · CPC title

• C07H1/00

  Processes for the preparation of sugar derivatives · CPC title

• B01J35/60

  characterised by their surface properties or porosity · CPC title

• B01J2229/32

  Reaction with silicon compounds, e.g. TEOS, siliconfluoride · CPC title

• B01J27/24

  Nitrogen compounds · CPC title