EP0035719A2
Expired / Lapsed
EP
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Process for producing modified inorganic polymers, their use in producing polynucleotides, and a reagent useful in these processes
Assignee: Competitive Technologies Inc
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Filing: 1981-02-27
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Granted: —
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Expires: 2001-02-27
Abstract
Abstract Modified inorganic polymers represented by the formula wherein Ⓟ is an inorganic polymer linked to the 3' or 5' -0- of the nucleoside through a base hydrolyzable covalent bond; R is H or a blocking group; R is a hydrocarbyl radical containing up to 10 carbons; each B is a nucleoside or deoxynucleoside base; and each A is H or OR. Said inorganic polymers are chemically bound to a nucleoside. The chemical bonding of the nucleoside moiety to the inorganic polymer is by means of reactive groups on the polymer which react with reactive groups of the nucleoside molecule. Such modified inorganic polymer supports are useful in the stepwise addition of nucleosides or oligonucleotides to the original nucleoside moiety of the support by a series of process steps. The starting nucleoside which is blocked in either the 3'- or the 5'-OH and the nucleoside is coupled through the unblocked hydroxy group to the polymer support bythe coupling agent. Furthermore an automated apparatus for producing a modified silica gel which is an example of a modified inorganic polymer is shown.
Inventors
- 👤 Marvin H. Caruthers
- 👤 Mark D. Matteucci
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-31 23:13
CPC Classifications
C; C07; C07H; C07H21/00; B; B01; B01J; B01J19/00; B01J19/0046; C07H19/00; C07H19/02; C07H19/04; C07H23/00; B01J2219/00; B01J2219/00274; B01J2219/00277; B01J2219/00279; B01J2219/00281; B01J2219/00286; B01J2219/00351; B01J2219/00389; B01J2219/00497; B01J2219/005; B01J2219/00583; B01J2219/0059; B01J2219/00596; B01J2219/0068; B01J2219/00686; B01J2219/00718; B01J2219/0072; B01J2219/00722; C40; C40B; C40B40/00; C40B40/04; C40B40/06; C40B60/00; C40B60/14; Y; Y02; Y02P; Y02P20/00; Y02P20/50; Y02P20/582
Claims Analysis
Total Claims: 32
Claim 1
Independent Claim
86 words
A process for production of oligonucleotides which comprises the step of condensing the 3' -OH or 5' -OH group of a nucleoside or oligonucleotide covalently linked to an inorganic polymer by a coupling agent through the 5'-0-or 3' -O-, respectively, of said nucleoside or oligonucleotide with a phosphite compound of the formula: wherein R is a blocking group; B is a nucleoside or deoxynucleoside base; A is H or OR; R 1 is lower alkyl; and X is Cl or Br, or a secondary amino group.
Claim 2
Dependent Claim
86 words
A process for production of oligonucleotides which comprises the step of condensing the 3' -OH or 5' -OH group of a nucleoside or oligonucleotide covalently linked to an inorganic polymer by a coupling agent through the 5'-0-or 3' -O-, respectively, of said nucleoside or oligonucleotide with a phosphite compound of the formula: wherein R is a blocking group; B is a nucleoside or deoxynucleoside base; A is H or OR; R 1 is lower alkyl; and X is Cl or Br, or a secondary amino group.
Claim 3
Dependent Claim
41 words
The process according to Claim 1 wherein the phosphite compound is of the formula: wherein substituents R, R 1 , X, A and B are as previously defined and said condensation occurs through the 5' -OH of said nucleoside or oligonucleotide.
Claim 4
Dependent Claim
12 words
The process according to Claim 2 wherein R 1 is lower alkyl.
Claim 5
Dependent Claim
15 words
.4.. The orocess accordinq to Claim 2 wherein the inorqanic polymer is a silica qel.
Claim 6
Dependent Claim
17 words
The process according to Claim 4 wherein A is H and R is a trityl blocking group.
Claim 7
Dependent Claim
23 words
The process according to Claim 5 wherein the free silanol hydroxy groups on the silica gel are blocked prior to the condensation reaction.
Claim 8
Dependent Claim
12 words
The process according to Claim 5 wherein the trityl group is dimethoxytrityl.
Claim 9
Dependent Claim
23 words
8-. The process according to Claim 4 wherein the silica gel is covalently linked to said nucleoside or oligonucleotide through an ester linkage.
Claim 10
Dependent Claim
20 words
The process according to Claim 4 including the further step of oxidizing the resulting phosphite triester to a phosphate triester.
Claim 11
Dependent Claim
13 words
The process according to Claim 3 wherein I 2 is the oxidizing agent.
Claim 12
Dependent Claim
34 words
The process according to Claim 9 wherein the 5' -0- blocking group of the product thus obtained is removed and'the product is then condensed with a further nucleoside phosphite compound of the said formula.
Claim 13
Dependent Claim
44 words
The process according to Claim 11 wherein (1) the product thus obtained is oxidized to obtain the corresponding phosphate triester; (2) the 5'-0- blocking group is removed; and (3) the product is condensed with a further nucleoside or oligonucleoside phosphite of the said formula.
Claim 14
Dependent Claim
28 words
The process according to Claim 12 wherein the 5' -0- blocking group is a trityl group and the said group is removed by reaction with a Lewis acid.
Claim 15
Dependent Claim
23 words
The pro:ess according to laim 13 wherein the 5' -0- blocking group is a dimethoxytrityl group and said Lewis acid is zinc bromide.
Claim 16
Dependent Claim
23 words
The pro:ess according to laim 13 wherein the 5' -0- blocking group is a dimethoxytrityl group and said Lewis acid is zinc bromide.
Claim 17
Dependent Claim
24 words
The process according to Claim 12 wherein each of steps 1, 2 and 3 is repeated to successively add nucleosides to the oligonucleotide chain.
Claim 18
Dependent Claim
27 words
The process according to Claim 15 wherein the oligonucleotide chain comprises from about 10 to about 30 nucleotides attached through the 5' -0 and 3' -0 positions.
Claim 19
Dependent Claim
98 words
A process for production of oligonucleotides which comprises the step of condensing a compound of the formula: . wherein B is a nucleoside or deoxynucleoside base, A is H , OH or OR; and Ⓟ is silica covalently bound to the nucleoside by the group -Z 1 NHCOZCO- wherein Z and Z 1 are each alkylene of up to 5 carbon atoms, with a nucleoside phosphite compound of the formula: wherein A and B are as previously defined; R 1 is lower alkyl; R is a blocking group; and X is Cl or Br, or a secondary amino.
Claim 20
Dependent Claim
98 words
A process for production of oligonucleotides which comprises the step of condensing a compound of the formula: . wherein B is a nucleoside or deoxynucleoside base, A is H , OH or OR; and Ⓟ is silica covalently bound to the nucleoside by the group -Z 1 NHCOZCO- wherein Z and Z 1 are each alkylene of up to 5 carbon atoms, with a nucleoside phosphite compound of the formula: wherein A and B are as previously defined; R 1 is lower alkyl; R is a blocking group; and X is Cl or Br, or a secondary amino.
Claim 21
Dependent Claim
19 words
The process according to Claim 17 including the further step of oxidizing the phosphite triester to a phosphate triester.
Claim 22
Dependent Claim
15 words
1 9. The process according to Claim 18 wherein I 2 is the oxidizing agent.
Claim 23
Dependent Claim
35 words
The process according to Claim 18 wherein the 5' -0- blocking group of the product thus obtained is removed and the product is then condensed with a further nucleoside phosphite compound of the said formula.
Claim 24
Dependent Claim
41 words
The process according to Claim20 wherein (1) the product thus obtained is oxidized to obtain the.corresponding phosphate triester; (2) the 5' -0 blocking group is removed; and (3) the product is condensed with a further nucleoside phosphite of the said formula.
Claim 25
Dependent Claim
24 words
The process according to Claim 21 wherein each of steps 1, 2 and 3 are repeated to successively add nucleosides to the oligonucleotide chain.
Claim 26
Dependent Claim
27 words
The process according to Claim 22 wherein the oligonucleotide chain comprises from about 10 to about 30 nucleosides attached through the 5' -0 and 3' -0 positions.
Claim 27
Dependent Claim
25 words
The process according to Claim22 wherein the phosphate triester product is hydrolyzed to form the oligonucleotide and silica gel, and the oligonucleotide is separated therefrom.
Claim 28
Dependent Claim
17 words
The process according to Claim 24 including the further step of removing blocking groups from the oligonucleotide.
Claim 29
Dependent Claim
160 words
An automated apparatus for producing a modified silica gel, comprising: a) first valve means (14) selectively connectable to a supply of active reagents or wash solvents; b) second valve means (28) selectively connectable to a supply nucleoside-active phosphate triester or a wash solvent; c) third valve means .(42) selectively connectable to a supply of cleavage reagents or a wash solvent; d) a collector valve (50) for receiving the outflow from, respectively, said first, second and third valve means; e) a closed flow circuit including a selector valve (52) ; a column (1C)containing a modified silica gel matrix (12); and pump means (56) arranged intermediate said selector valve (52)and said column in series connection; said collector valve (50) having the outflow thereof connected to an inlet of said selector valve (52); f) and a uv detector (58) connected to an outlet of said selector valve (52) for conveying said modified silica gel (12) to either a collector or a waste arrangement.
Claim 30
Dependent Claim
160 words
An automated apparatus for producing a modified silica gel, comprising: a) first valve means (14) selectively connectable to a supply of active reagents or wash solvents; b) second valve means (28) selectively connectable to a supply nucleoside-active phosphate triester or a wash solvent; c) third valve means .(42) selectively connectable to a supply of cleavage reagents or a wash solvent; d) a collector valve (50) for receiving the outflow from, respectively, said first, second and third valve means; e) a closed flow circuit including a selector valve (52) ; a column (1C)containing a modified silica gel matrix (12); and pump means (56) arranged intermediate said selector valve (52)and said column in series connection; said collector valve (50) having the outflow thereof connected to an inlet of said selector valve (52); f) and a uv detector (58) connected to an outlet of said selector valve (52) for conveying said modified silica gel (12) to either a collector or a waste arrangement.
Claim 31
Dependent Claim
38 words
The apparatus of Claim 26 comprising programmable controller means (15, 15', 15") connected to each of said first, second and third valve means (14, 28, 42) for determining the outflow of said valve means to said collector valve.
Claim 32
Dependent Claim
36 words
The apparatus of Claim 26 said selector valve being adjustable to direct flow from said collector valve (50) through said column (10) to said uv detector (58) or alternatively, to recycle flow through said column (10).
References Cited (Backward Citations)
No citation records indexed.
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