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US6426408B1 Expired / Lapsed US ⏱️ 0.0 years remaining

Covalently linked oligonucleotide minor groove binder conjugates

Assignee: DRUG ROYALTY TRUST 9
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Filing: 2000-02-18
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Granted: 2002-07-30
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Expires: 2020-02-18
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Abstract

Abstract Minor groove binding molecules are covalently bound to oligonucleotides which in their base sequence are complementary to a target sequence of single stranded or double stranded DNA, RNA or hybrids thereof. The covalently bound oligonucleotide minor groove binder conjugates strogly bind to the target sequence of the complementary strand.

Inventors

  • 👤 Igor V. Kutyavin
  • 👤 Eugeny A. Lukhtanov
  • 👤 Howard B. Gamper
  • 👤 Rich B. Meyer, Jr.

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-29 18:31
CPC Classifications
C; C07; C07H; C07H21/00; C07D; C07D207/00; C07D207/02; C07D207/30; C07D207/34; C07D403/00; C07D403/02; C07D403/12; C07D487/00; C07D487/02; C07D487/04; C07H19/00; C07H19/02; C07H19/04; C07H19/06; C07H19/073; C07H19/16; C07H19/173; C07H19/23; C07H21/04; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813

Claims Analysis

Total Claims: 8
Claim 1
Independent Claim 653 words
An oligonucleotide minor groove binder combination comprising an oligonucleotide having a plurality of nucleotide units, a 3′-end and a 5′-end, and a minor groove binder moiety attached to at least one of said nucleotides through a linking group which covalently binds the minor groove binder moiety to the oligonucleotide, wherein the minor groove binder moiety is a radical of a molecule having a molecular weight of approximately 150 to approximately 2000 Daltons that binds in a non-intercalating manner into the minor groove of double stranded DNA, RNA or hybrids thereof with an association constant greater than approximately 10 3 M −1 ; wherein the minor groove binding moiety including the linking group is represented by the formula: R 1 —(R 6 N—Y 2 —CO) n —R 2 wherein Y 2 is a ring system consisting of a 6-membered aromatic ring condensed with a 5-membered ring having one double bond, the condensed ring system having 0 to 3 heteroatoms selected from the group consisting of N, S and O, each of the R 6 N and CO groups is attached to a ring carbon which is in a different ring of the condensed ring system, and which is the second ring atom, respectively, from one common bridgehead ring atom, the CO and NR 6 groups thereby positioning 2 non-bridgehead ring atoms between themselves on one side and 3 non-bridgehead ring atoms on the other side of the condensed ring system, the two non-bridgehead ring atoms on the one side being optionally substituted with an R 7 group, the three non-bridgehead ring atoms on the other side of the condensed ring system being optionally substituted with an R 3 group; wherein one of R 1 or R 2 is a linking group forming a covalent attachment to said oligonucleotide and the other of R 1 or R 2 is selected from the group consisting of H, F, Cl, Br, I, NH 2 , NHR 4 , N(R 4 ) 2 , N(R 4 ) 3 + , OH, OR 4 , SH, SR 4 , COR 4 , CONHR 4 , CON(R 4 ) 2 , R 4 , H 2 N(CH 2 ) m CO, CONH 2 , CONHR 4 and H 2 N(CH 2 ) m COO (CH 2 ) m S(CH 2 ) m C 6 H 4 NNC 6 H 4 ; R 3 is selected from the group consisting of F, Cl, Br, I, NH 2 , NHR 4 , N(R 4 ) 2 , N(R 4 ) 3 + , OH, OR 4 , SH, SR 4 , COR 4 , CONHR 4 , CON(R 4 ) 2 and R 4 , or the R 3 groups may form a 3, 4, 5 or 6 membered ring condensed to the Y 2 ring and substituted by one, two or three R 4 ; R 4 is an alkyl or cycloalkyl group having 1 to 20 carbons, an alkenyl or cycloalkenyl group having 1 to 20 carbons and 1 to 3 double bonds, a carbocyclic aromatic group of no more than 25 carbons, a heterocyclic aromatic group of no more than 25 carbons, a carbocyclic or heterocyclic arylalkyl group of no more than 25 carbons, where R 4 may be optionally substituted with 1, 2 or 3 F, Cl, Br, I, NH 2 , NHR 5 , N(R 5 ) 2 , N(R 5 ) 3 + , OH, OR 5 , SH, SR 5 , COR 5 , CONHR 5 , CON(R 5 ) 2 or R 5 groups; R 5 is alkyl of 1 to 6 carbons, R 6 and R 7 jointly form a 4, 5, or 6 membered ring, optionally an —O—, —S—, —NH—, —NCH 3 —, or N -lower alkyl group being part of said ring; m is an integer between 1 and 10; and n is an integer between of from 2 and 5.
Claim 2
Dependent Claim 653 words
An oligonucleotide minor groove binder combination comprising an oligonucleotide having a plurality of nucleotide units, a 3′-end and a 5′-end, and a minor groove binder moiety attached to at least one of said nucleotides through a linking group which covalently binds the minor groove binder moiety to the oligonucleotide, wherein the minor groove binder moiety is a radical of a molecule having a molecular weight of approximately 150 to approximately 2000 Daltons that binds in a non-intercalating manner into the minor groove of double stranded DNA, RNA or hybrids thereof with an association constant greater than approximately 10 3 M −1 ; wherein the minor groove binding moiety including the linking group is represented by the formula: R 1 —(R 6 N—Y 2 —CO) n —R 2 wherein Y 2 is a ring system consisting of a 6-membered aromatic ring condensed with a 5-membered ring having one double bond, the condensed ring system having 0 to 3 heteroatoms selected from the group consisting of N, S and O, each of the R 6 N and CO groups is attached to a ring carbon which is in a different ring of the condensed ring system, and which is the second ring atom, respectively, from one common bridgehead ring atom, the CO and NR 6 groups thereby positioning 2 non-bridgehead ring atoms between themselves on one side and 3 non-bridgehead ring atoms on the other side of the condensed ring system, the two non-bridgehead ring atoms on the one side being optionally substituted with an R 7 group, the three non-bridgehead ring atoms on the other side of the condensed ring system being optionally substituted with an R 3 group; wherein one of R 1 or R 2 is a linking group forming a covalent attachment to said oligonucleotide and the other of R 1 or R 2 is selected from the group consisting of H, F, Cl, Br, I, NH 2 , NHR 4 , N(R 4 ) 2 , N(R 4 ) 3 + , OH, OR 4 , SH, SR 4 , COR 4 , CONHR 4 , CON(R 4 ) 2 , R 4 , H 2 N(CH 2 ) m CO, CONH 2 , CONHR 4 and H 2 N(CH 2 ) m COO (CH 2 ) m S(CH 2 ) m C 6 H 4 NNC 6 H 4 ; R 3 is selected from the group consisting of F, Cl, Br, I, NH 2 , NHR 4 , N(R 4 ) 2 , N(R 4 ) 3 + , OH, OR 4 , SH, SR 4 , COR 4 , CONHR 4 , CON(R 4 ) 2 and R 4 , or the R 3 groups may form a 3, 4, 5 or 6 membered ring condensed to the Y 2 ring and substituted by one, two or three R 4 ; R 4 is an alkyl or cycloalkyl group having 1 to 20 carbons, an alkenyl or cycloalkenyl group having 1 to 20 carbons and 1 to 3 double bonds, a carbocyclic aromatic group of no more than 25 carbons, a heterocyclic aromatic group of no more than 25 carbons, a carbocyclic or heterocyclic arylalkyl group of no more than 25 carbons, where R 4 may be optionally substituted with 1, 2 or 3 F, Cl, Br, I, NH 2 , NHR 5 , N(R 5 ) 2 , N(R 5 ) 3 + , OH, OR 5 , SH, SR 5 , COR 5 , CONHR 5 , CON(R 5 ) 2 or R 5 groups; R 5 is alkyl of 1 to 6 carbons, R 6 and R 7 jointly form a 4, 5, or 6 membered ring, optionally an —O—, —S—, —NH—, —NCH 3 —, or N -lower alkyl group being part of said ring; m is an integer between 1 and 10; and n is an integer between of from 2 and 5.
Claim 3
Dependent Claim 26 words
An oligonucleotide minor groove binder combination in accordance with claim 1 , wherein the minor groove binding moiety is attached to the 5′-end of the oligonucleotide.
Claim 4
Dependent Claim 26 words
An oligonucleotide minor groove binder combination in accordance with claim 1 , wherein the minor groove binding moiety is attached to the 3′-end of the oligonucleotide.
Claim 5
Dependent Claim 38 words
An oligonucleotide minor groove binder combination in accordance with claim 1 , wherein the minor groove binder moiety is attached to a nucleotide unit which is neither at the 3′ nor at the 5′ end of the oligonucleotide.
Claim 6
Dependent Claim 29 words
An oligonucleotide minor groove binder combination in accordance with claim 1 , wherein the minor groove binder moiety is attached to the heterocyclic base portion of a nucleotide unit.
Claim 7
Dependent Claim 261 words
An oligonucleotide minor groove binder combination in accordance with claim 1 , the combination having the formula wherein X is O or S; q is an integer between 3 to 100; each R 8 is independently selected from the group consisting of H, OH, alkoxy having 1 to 6 carbons, O—C 2 —C 6 alkenyl, and F; B is an aglycone selected from a group consisting of a heterocyclic base naturally found in nucleic acids, hypoxanthine, 2-aminoadenine, 2-thiouracil, 2-thiothymine, 5- N 4 -ethenocytosine, 4-aminopyrrazolo [3,4- d ] pyrimidine, and 6-amino-4-hydroxy-[3,4- d ] pyrimidine; W 1 is H, PO(OH) 2 or a salt thereof, or a minor groove binder moiety attached to the 3′ or 5′ end of said oligonucleotide, including the linking group which covalently binds the minor groove binder moiety to the oligonucleotide; W 2 is absent or is a minor groove binder moiety attached to one of the aglycones B including the linking group which covalently binds the minor groove binder moiety to said aglycone, or W 2 is a cross-linking functionality including a linker arm which covalently binds the cross-linking functionality to said aglycone; wherein the minor groove binder moiety is a radical of a molecule having a molecular weight of approximately 150 to approximately 2000 Daltons that binds in a non-intercalataing manner into the minor groove of double stranded DNA, RNA or hybdrids thereof with an association constant greater than approximately 10 3 M −1 , with the proviso that at least one of said W 1 and W 2 groups comprises a minor groove binder moiety.
Claim 8
Dependent Claim 15 words
An oligonucleotide minor groove binder combination of claim 1 , further comprising a reporter group.

References Cited (Backward Citations)

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US5237101A Cited
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US5395849A Cited
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3.5 Years (Year 4) 2005-07-29 2006-01-28 2006-07-30 $2000.00 Window Closed / Lapsed
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