EP0313219A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Non-nucleotide linking reagents for nucleotide probes
Assignee: Gen Probe Inc
•
Filing: 1988-09-21
•
Granted: —
•
Expires: 2008-09-21
Abstract
Abstract A versatile reagent with a non-nucleotide monomeric unit having a ligand, and first and second coupling groups which are linked to the non-nucleotide monomeric unit. The ligand can be either a functional moiety, such as a label or intercalator, or a linking arm which can be linked to such a moiety. Such reagent permits preparation of versatile nucleotide/non-nucleotide polymers, having any desired sequence of nucleotide and non-nucleotide monomeric units, each of the latter of which bear a desired ligand. These polymers can for example, be used as probes which can exhibit enhanced sensitivity and/or which are capable of detecting a genus of nucleotides each species of which has a common target nucleotide sequence of interest bridged by different sequences not of interest.
Inventors
- 👤 Lyle John Arnold
- 👤 Mark Alan Reynolds
- 👤 Ram Saroop Bhatt
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-06 05:22
CPC Classifications
C; C07; C07H; C07H21/00; C07H21/02; C07F; C07F9/00; C07F9/02; C07F9/06; C07F9/22; C07F9/24; C07F9/2404; C07F9/2408
Claims Analysis
Total Claims: 51
Claim 1
Independent Claim
131 words
A non-nucleotide reagent suitable for preparing a nucleotide/non-nucleotide polymer, comprising: (a) a non-nucleotide monomeric unit, having a ligand which is selected from a chemical moiety, and from a linking arm which can be activated under non-adverse conditions so as to be capable of linking with a chemical moiety; (b) first and second non-nucleotide coupling groups linked to the monomeric unit, the first coupling group of which is capable of coupling the non-nucleotide monomeric unit to a first additional monomeric unit, while the second coupling group remains inactivated so as to be substantially incapable of coupling, but can thereafter be activated under non-adverse conditions to couple the non-nucleotide monomeric unit to a second additional monomeric unit, at least one of the first and second additional monomeric units being a nucleotide monomeric unit.
Claim 2
Dependent Claim
131 words
A non-nucleotide reagent suitable for preparing a nucleotide/non-nucleotide polymer, comprising: (a) a non-nucleotide monomeric unit, having a ligand which is selected from a chemical moiety, and from a linking arm which can be activated under non-adverse conditions so as to be capable of linking with a chemical moiety; (b) first and second non-nucleotide coupling groups linked to the monomeric unit, the first coupling group of which is capable of coupling the non-nucleotide monomeric unit to a first additional monomeric unit, while the second coupling group remains inactivated so as to be substantially incapable of coupling, but can thereafter be activated under non-adverse conditions to couple the non-nucleotide monomeric unit to a second additional monomeric unit, at least one of the first and second additional monomeric units being a nucleotide monomeric unit.
Claim 3
Dependent Claim
64 words
A reagent as defined in claim 1, wherein said ligand is selected from a label, an intercalator, a metal chelator, drugs, hormones, proteins, peptides, haptens, radical generators, nucleolytic agents, proteolytic agents, catalysts, specific binding substances, agents which modify DNA transport, and substances which alter nucleotide multimer solubility, and from a linking functional group which can be linked to any of the foregoing chemical moieties.
Claim 4
Dependent Claim
134 words
A non-nucleotide reagent suitable for preparing a nucleotide/non-nucleotide polymer, comprising: (a) a non-nucleotide monomeric unit, having a ligand which is selected from a chemical moiety, and from a protected linking arm which can be deprotected under non-adverse conditions so as to be capable of linking with a chemical moiety; (b) a non-nucleotide, first coupling group and protected second coupling group, linked to the monomeric unit, the first coupling group of which is capable of coupling the non-nucleotide monomeric unit to a first additional monomeric unit while the second one is substantially incapable of coupling, but can thereafter be deprotected under non-adverse conditions, so as to be capable of coupling the non-nucleotide monomeric unit to a second additional monomeric unit, at least one of the first and second monomeric units being a nucleotide monomeric unit.
Claim 5
Dependent Claim
134 words
A non-nucleotide reagent suitable for preparing a nucleotide/non-nucleotide polymer, comprising: (a) a non-nucleotide monomeric unit, having a ligand which is selected from a chemical moiety, and from a protected linking arm which can be deprotected under non-adverse conditions so as to be capable of linking with a chemical moiety; (b) a non-nucleotide, first coupling group and protected second coupling group, linked to the monomeric unit, the first coupling group of which is capable of coupling the non-nucleotide monomeric unit to a first additional monomeric unit while the second one is substantially incapable of coupling, but can thereafter be deprotected under non-adverse conditions, so as to be capable of coupling the non-nucleotide monomeric unit to a second additional monomeric unit, at least one of the first and second monomeric units being a nucleotide monomeric unit.
Claim 6
Dependent Claim
64 words
A reagent as defined in claim 3, wherein said ligand is selected from a label, an intercalator, a metal chelator, drugs, hormones, proteins, peptides, haptens, radical generators, nucleolytic agents, proteolytic agents, catalysts, specific binding substances, agents which modify DNA transport, and substances which alter nucleotide multimeter solubility, and from a linking functional group which can be linked to any of the foregoing chemical moieties.
Claim 7
Dependent Claim
35 words
A reagent as defined in claim 4 wherein the first and second coupling groups, are capable of coupling the non-nucleotide monomeric unit to a 5′ hydroxyl and a 3′ phosphate of a monomeric unit, respectively.
Claim 8
Dependent Claim
29 words
A reagent as defined in claim 5 wherein the non-nucleotide monomeric unit has an acyclic skeleton to respective ends of which the first and second coupling groups are linked.
Claim 9
Dependent Claim
37 words
A reagent as defined in claim 5 wherein the non-nucleotide monomeric unit has a skeleton to respective ends of which the first and second coupling groups are linked, the skeleton being a 1 to 20 atom chain.
Claim 10
Dependent Claim
39 words
A reagent as defined in claim 5 wherein the non-nucleotide monomeric unit has a skeleton to respective ends of which are linked the coupling groups, the skeleton being an acyclic hydrocarbon chain of from 1 to 10 carbon atoms.
Claim 11
Dependent Claim
41 words
A reagent as defined in claim 5 wherein the non-nucleotide monomeric unit has a skeleton to respective ends of which are linked the coupling groups, and wherein the linking arm is a 1 to 25 atom chain extending from the skeleton.
Claim 12
Dependent Claim
107 words
A reagent as defined in claim 2 selected from one of the compounds having the formula: wherein: (a) R₂ = a skeleton of the non-nucleotide monomeric unit; (b) are first coupling groups in which: X₂ = halogen or substituted amino X₄ = halogen; amino; or O⁻ R₃ = alkyl; alkoxy; or phenoxy R5 = alkyl; alkoxy; or aryloxy or may be H only if X4 = O⁻ (c) R₁-X₁- is the protected second coupling group in which: X₁ = O; S; NH; or NH=N- R₁ = the protecting group cleavable under coupling group deprotecting conditions to recover the second coupling group H-X₁-; (d) n is an integer;
Claim 13
Dependent Claim
46 words
A reagent as defined in claim 10 wherein: X₂ = Cl; or secondary amino R₃ = chlorophenoxy;methoxy;ethoxy; or beta-cyanoethoxy X4 = Cl; secondary amino; or O⁻ when R₃ = H R5 = methoxy; ethoxy; monochlorophenoxy; or beta-cyanoethoxy; or may be H only if X4 = O⁻
Claim 14
Dependent Claim
42 words
A reagent as defined in claim 11 wherein the ligand is a protected linking arm in which X3 is an alkylamino linked to R2 by a C- and to R4 by N-, and in which R4 is selected from trifluoroacetyl and 9-fluoremylmethoxycarbonyl.
Claim 15
Dependent Claim
17 words
A reagent as defined in claim 11 wherein R₂ has a secondary carbon linked to the -O-.
Claim 16
Dependent Claim
19 words
A reagent as defined in claim 10, wherein the secondary amino groups are selected from dialkylamino, and heterocyclic N-amines.
Claim 17
Dependent Claim
19 words
A reagent as defined in claims 10, 11 or 14, wherein R₁ = triphenylmethyl or an alkoxy derivative thereof.
Claim 18
Dependent Claim
23 words
A reagent as defined in claim 10, 11 and 14, wherein X₁ = O and R₁ - triphenylmethyl or an alkoxy derivative thereof.
Claim 19
Dependent Claim
18 words
A reagent as defined in claim 10, 11 or 14 wherein X₁ = O and R₁ = dimethoxytriphenylmethyl.
Claim 20
Independent Claim
76 words
A method of preparing a substituted nucleotide from a reagent having: (a) a non-nucleotide monomeric unit; and (b) a ligand linked to the non-nucleotide monomeric unit, and selected from a chemical moiety, and from a linking arm to which a chemical moiety can be linked under non-adverse conditions; the method comprising coupling the non-nucleotide monomeric unit under non-adverse conditions, to a nucleotide monomeric unit and to one of a second monomeric unit and a solid support.
Claim 21
Independent Claim
76 words
A method of preparing a substituted nucleotide from a reagent having: (a) a non-nucleotide monomeric unit; and (b) a ligand linked to the non-nucleotide monomeric unit, and selected from a chemical moiety, and from a linking arm to which a chemical moiety can be linked under non-adverse conditions; the method comprising coupling the non-nucleotide monomeric unit under non-adverse conditions, to a nucleotide monomeric unit and to one of a second monomeric unit and a solid support.
Claim 22
Independent Claim
134 words
A method of preparing a substituted nucleotide from a reagent having: i) a non-nucleotide monomeric unit having a ligand, selected from a functional moiety and from a protected linking arm to which a functional moiety can be linked; and ii) a first coupling group and a protected second coupling group linked to the non-nucleotide monomeric unit; the method comprising, under non-adverse conditions: a) first coupling the non-nucleotide monomeric unit to a first additional monomeric unit through the first coupling group; b) then deprotecting the second coupling group; c) then coupling the non-nucleotide monomeric unit through the second coupling group, to a second additional monomeric unit; at least one of the first and second monomeric units being a nucleotide monomeric unit while the other one is selected from a monomeric unit and a solid support.
Claim 23
Independent Claim
134 words
A method of preparing a substituted nucleotide from a reagent having: i) a non-nucleotide monomeric unit having a ligand, selected from a functional moiety and from a protected linking arm to which a functional moiety can be linked; and ii) a first coupling group and a protected second coupling group linked to the non-nucleotide monomeric unit; the method comprising, under non-adverse conditions: a) first coupling the non-nucleotide monomeric unit to a first additional monomeric unit through the first coupling group; b) then deprotecting the second coupling group; c) then coupling the non-nucleotide monomeric unit through the second coupling group, to a second additional monomeric unit; at least one of the first and second monomeric units being a nucleotide monomeric unit while the other one is selected from a monomeric unit and a solid support.
Claim 24
Dependent Claim
22 words
A method as defined in claim 18 or 19, wherein both the first and second additional monomeric units are nucleotide monomeric units.
Claim 25
Dependent Claim
36 words
A method as defined in claim 18 wherein the first and second coupling groups, are capable of coupling the non-nucleotide monomeric unit to a 5′ hydroxyl and a 3′ phosphate of a nucleotide monomeric unit, respectively.
Claim 26
Dependent Claim
29 words
A method as defined in claim 19 wherein the non-nucleotide monomeric unit has an acyclic skeleton to respective ends of which the first and second coupling groups are linked.
Claim 27
Dependent Claim
22 words
A method as defined in claim 19 wherein the reagent is one as defined in any one of claims 10 to 18.
Claim 28
Dependent Claim
41 words
A method as defined in claim 23, wherein said ligand is selected from a label, an intercalator, a metal chelator, and from a linking functional group which can be linked to one of a lable, an intercalator, and a metal chelator.
Claim 29
Dependent Claim
65 words
A method as defined in claim 23 or 24 additionally comprising coupling further monomeric units to the coupled nucleotide/non-nucleotide polymer, to produce a polymer with a nucleotide sequence which is complementary to at least a portion of a target nucleotide sequence, but which complementary nucleotide sequence has at least one non-nucleotide monomeric unit disposed therein, then hybridizing the nucleotide/non-nucleotide polymer sequence with the target sequence.
Claim 30
Dependent Claim
76 words
A composition containing a single sequence nucleotide/non-nucleotide polymer as the majority of all nucleotide/non-nucleotide polymers present, the single nucleotide/non-nucleotide polymer comprising: a) a non-nucleotide monomeric unit having linked thereto a ligand selected from a functional moiety, and from a linking arm which can be linked to a functional moiety under non-adverse conditions; b) a first nucleotide monomeric unit coupled to the non-nucleotide monomeric unit; and c) an additional monomeric unit coupled to the non-nucleotide monomeric unit.
Claim 31
Dependent Claim
76 words
A composition containing a single sequence nucleotide/non-nucleotide polymer as the majority of all nucleotide/non-nucleotide polymers present, the single nucleotide/non-nucleotide polymer comprising: a) a non-nucleotide monomeric unit having linked thereto a ligand selected from a functional moiety, and from a linking arm which can be linked to a functional moiety under non-adverse conditions; b) a first nucleotide monomeric unit coupled to the non-nucleotide monomeric unit; and c) an additional monomeric unit coupled to the non-nucleotide monomeric unit.
Claim 32
Dependent Claim
41 words
A composition as defined in claim 26, wherein said ligand is selected from a label, an intercalator, a metal chelator, and from a linking functional group which can be linked to one of a label, an intercalator and a metal chelator.
Claim 33
Dependent Claim
20 words
A composition as defined in either of claim 26 or 27 wherein the additional unit is a nucleotide monomeric unit.
Claim 34
Dependent Claim
29 words
A composition as defined in either of claim 26 or 27 wherein the additional monomeric unit is a non-nucleotide monomeric unit, which is coupled to another non-nucleotide monomeric unit.
Claim 35
Dependent Claim
71 words
A composition containing a nucleotide/non-nucleotide polymer comprising: a) a non-nucleotide monomeric unit, having linked thereto a ligand selected from a functional moiety linked to the skeleton through a saturated linker group, and from a saturated linking arm which can be linked to a ligand under non-adverse conditions; b) a first nucleotide monomeric unit coupled to the non-nucleotide monomeric unit; and c) an additional monomeric unit coupled to the non-nucleotide monomeric unit.
Claim 36
Dependent Claim
71 words
A composition containing a nucleotide/non-nucleotide polymer comprising: a) a non-nucleotide monomeric unit, having linked thereto a ligand selected from a functional moiety linked to the skeleton through a saturated linker group, and from a saturated linking arm which can be linked to a ligand under non-adverse conditions; b) a first nucleotide monomeric unit coupled to the non-nucleotide monomeric unit; and c) an additional monomeric unit coupled to the non-nucleotide monomeric unit.
Claim 37
Dependent Claim
38 words
A composition as defined in claim 30, wherein said ligand is selected from a label, an intercalator, a metal chelator, and from a saturated linking arm which can be linked to either of a label and an intercalator.
Claim 38
Dependent Claim
18 words
A composition as defined in claim 30 or 31 wherein the ligand group is the saturated linking arm.
Claim 39
Dependent Claim
48 words
A composition as defined in claim 29 wherein the non-nucleotide monomeric unit has a skeleton to the ends of which are linked the coupling groups, and wherein the ligand comprises a label linked to the skeleton through the linking group which is selected from an alkylamino and alkylthio.
Claim 40
Dependent Claim
19 words
A composition as defined in claim 32 wherein the linking arm is selected from an alkylamino and an alkylthio.
Claim 41
Dependent Claim
42 words
A composition as defined in claim 30, 32 or 34 wherein the non-nucleotide monomeric unit has a skeleton to the ends of which are linked the coupling groups, the skeleton being an acyclic hydrocarbon chain having from 1 to 20 carbon atoms.
Claim 42
Dependent Claim
15 words
A composition as defined in claim 30 wherein the label is a chemiluminescent acridinium ester.
Claim 43
Dependent Claim
14 words
A composition as defined in claim 31 wherein the intercalator is an acridinium ester.
Claim 44
Dependent Claim
20 words
A composition as defined in claim 30 wherein the label is selected from biotin, fluorescein, dinitrobenzene, rhodamine, and Texas Red.
Claim 45
Dependent Claim
20 words
A composition as defined in claim 30 wherein the intercalator is selected from psoralen, acridine, acridinium salts, acriflavins, and ethidium.
Claim 46
Dependent Claim
26 words
A polymeric probe comprising a plurality of nucleotide monomeric units, and at least one acridinium ester moiety linked to a corresponding monomeric unit of the probe.
Claim 47
Dependent Claim
26 words
A polymeric probe comprising a plurality of nucleotide monomeric units, and at least one acridinium ester moiety linked to a corresponding monomeric unit of the probe.
Claim 48
Dependent Claim
18 words
A probe as defined in claim 40 wherein the acridinium ester label is a chemiluminescent acridinium ester label.
Claim 49
Independent Claim
29 words
A method of making a probe, comprising linking at least one acridinium ester moiety to a corresponding monomeric unit of a polymer having a plurality of nucleotide monomeric units.
Claim 50
Independent Claim
29 words
A method of making a probe, comprising linking at least one acridinium ester moiety to a corresponding monomeric unit of a polymer having a plurality of nucleotide monomeric units.
Claim 51
Dependent Claim
18 words
A method as defined in claim 42 wherein the acridinium ester label is a chemiluminescent acridinium ester label.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.