Hydrazine-based and carbonyl-based bifunctional crosslinking reagents
First Claim
Patent Images
1. A compound of formula I:
- B—
R—
A—
NHNH2.HX
Ior a derivative thereof, wherein;
A is —
NH(C═
O)—
, —
NH(C═
S)—
, —
NHNH(C═
O)—
, —
NHNH(C═
S)—
, or a direct bond to R;
B is an amino or thiol reactive moiety;
R is an aliphatic divalent group having any combination of the following groups, which are combined in any order;
cycloalkylene, C(R10)2, —
C(R10)═
C(R10)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G)a, P(J)b(R10), P(J)b(LR10), N(R10), >
N+(R12)(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G is O or NR10;
J is S or O; and
L is S, O or NR10;
each R10 is a monovalent group independently selected from hydrogen and M1-R14;
each M1 is a divalent group independently having any combination of the following groups, which groups are combined in any order;
a direct link, arylene, heteroarylene, cycloalkylene, C(R15)2, —
C(R15)═
C(R15)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G1)a, P(J)b(R15), P(J)b(LR15), N(R15), N(COR15), >
N+(R12)(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G1 is O or NR15;
J is S or O; and
L is S, O or NR15;
R14 and R15 are each independently selected from the group among hydrogen, halo, pseudohalo, cyano, azido, nitro, SiR16R17R18, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20;
R19 and R20 are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl and heterocyclyl;
R12 and R13 are selected from (i) or (ii) as follows;
(i) R12 and R13 are independently selected from among hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl and heteroaryl;
or (ii) R12 and R13 together form alkylen, alkenylene or cycloalkylene;
R16, R17 and R18 are each independently a monovalent group sel cted from hydrogen, alkyl, alk nyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20; and
R11, R12, R13, R14, R15, R16, R17, R18, R19 and R20 can be substituted with one or more substituents each independently selected from Z, wherein Z is selected from alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, hydroxy, S(O)hR30, NR30R31, COOR30, COR30, CONR30R31, OC(O)NR30R31, N(R30)C(O)R31, alkoxy, aryloxy, heteroaryl, heterocyclyl, heteroaryloxy, heterocyclyloxy, aralkyl, aralkenyl, aralkynyl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, aralkoxy, heteroaralkoxy, alkoxycarbonyl, carbamoyl, thiocarbamoyl, alkoxycarbonyl, carboxyaryl, halo, pseudohalo, haloalkyl and carboxamido;
h is 0, 1 or 2; and
R30 and R31 are each independently selected from among hydrogen, halo, pseudohalo, cyano, azido, nitro, trialkylsilyl, dialkylarylsilyl, alkyldiarylsilyl, triarylsilyl, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy, amino, amido, alkylamino, dialkylamino, alkylarylamino, diarylamino and arylamino; and
X is a negative counterion.
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Accused Products
Abstract
Reagents and methods are provided for crosslinking and immobilizing biomolecules, drugs and synthetic polymers. The reagents possess (i) a thiol or amino reactive group; and (ii) a hydrazino or oxyamino moiety. Conjugates and immobilized biomolecules are also provided.
-
Citations
53 Claims
-
1. A compound of formula I:
-
B—
R—
A—
NHNH2.HX
Ior a derivative thereof, wherein;
A is —
NH(C═
O)—
, —
NH(C═
S)—
, —
NHNH(C═
O)—
, —
NHNH(C═
S)—
, or a direct bond to R;
B is an amino or thiol reactive moiety;
R is an aliphatic divalent group having any combination of the following groups, which are combined in any order;
cycloalkylene, C(R10)2, —
C(R10)═
C(R10)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G)a, P(J)b(R10), P(J)b(LR10), N(R10), >
N+(R12)(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G is O or NR10;
J is S or O; and
L is S, O or NR10;
each R10 is a monovalent group independently selected from hydrogen and M1-R14;
each M1 is a divalent group independently having any combination of the following groups, which groups are combined in any order;
a direct link, arylene, heteroarylene, cycloalkylene, C(R15)2, —
C(R15)═
C(R15)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G1)a, P(J)b(R15), P(J)b(LR15), N(R15), N(COR15), >
N+(R12)(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G1 is O or NR15;
J is S or O; and
L is S, O or NR15;
R14 and R15 are each independently selected from the group among hydrogen, halo, pseudohalo, cyano, azido, nitro, SiR16R17R18, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20;
R19 and R20 are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl and heterocyclyl;
R12 and R13 are selected from (i) or (ii) as follows;
(i) R12 and R13 are independently selected from among hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl and heteroaryl;
or (ii) R12 and R13 together form alkylen, alkenylene or cycloalkylene;
R16, R17 and R18 are each independently a monovalent group sel cted from hydrogen, alkyl, alk nyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20; and
R11, R12, R13, R14, R15, R16, R17, R18, R19 and R20 can be substituted with one or more substituents each independently selected from Z, wherein Z is selected from alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, hydroxy, S(O)hR30, NR30R31, COOR30, COR30, CONR30R31, OC(O)NR30R31, N(R30)C(O)R31, alkoxy, aryloxy, heteroaryl, heterocyclyl, heteroaryloxy, heterocyclyloxy, aralkyl, aralkenyl, aralkynyl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, aralkoxy, heteroaralkoxy, alkoxycarbonyl, carbamoyl, thiocarbamoyl, alkoxycarbonyl, carboxyaryl, halo, pseudohalo, haloalkyl and carboxamido;
h is 0, 1 or 2; and
R30 and R31 are each independently selected from among hydrogen, halo, pseudohalo, cyano, azido, nitro, trialkylsilyl, dialkylarylsilyl, alkyldiarylsilyl, triarylsilyl, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy, amino, amido, alkylamino, dialkylamino, alkylarylamino, diarylamino and arylamino; and
X is a negative counterion. - View Dependent Claims (2, 3, 4, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 41, 42, 43, 44)
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-
4. The compound of claim 1 that is:
-
22. The compound of claim 1, wherein X is a halide or trifluoroacetate.
-
23. The compound of claim 1, wherein B is an amino reactive moiety selected from succininimidyl ester, hydroxybenzotriazolyl ester, or pentafluorophenol ester.
-
24. The compound of claim 1, wherein B is a thiol reactiv moiety selected from maleimido, α
- -bromoacetyl or pyridyidisulfide.
-
25. A conjugate, comprising the compound of claim 1 bound to a natural or synthetic biological molecule.
-
26. The conjugate of claim 25, wherein the natural or synthetic molecule is selected from a protein, a glycoprotein, a peptide, an oligonucleotide, an RNA, a DNA and a synthetic polymer.
-
27. The conjugate of claim 26, wherein the protein is an antibody.
-
28. A method of immobilizing a natural or synthetic biological molecule, comprising:
-
(i) preparing the conjugate of claim 25;
and(ii) applying the conjugate to a surface wherein the surface has at least one carbonyl moiety for a time and under conditions such that the hydrazine moiety of the conjugate reacts with the carbonyl moiety of the surface forming a hydrazone bond to the surface.
-
-
29. A method of crosslinking, a natural or synthetic biological molecule, comprising:
-
(i) preparing the conjugate of claim 25;
and(ii) applying the conjugate to a surface wherein the surface has at least one amino or one thiol reactive moiety for a time and under conditions such that the conjugate reacts with the amino moiety or thiol moiety of the surface forming a bond to the surface.
-
-
30. A method of crosslinking a natural or synthetic biological molecule, comprising:
-
(i) preparing the conjugate of claim 25;
and(ii) mixing the conjugate with a natural or synthetic biological molecule wherein the molecule has at least one carbonyl moiety for a time and under conditions such that the hydrazine moiety of the conjugate reacts with the carbonyl moiety of the molecule forming a hydrazone bond to the molecule.
-
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40. The method of claim 28, wherein the surface is selected from glass, polymer, latex and colloidal metal.
-
41. The method of claim 30, wherein the natural or synthetic biological molecule is selected from a protein, a glycoprotein, a peptide, an oligonucleotide, an RNA and a DNA.
-
42. The method of claim 41, wher in the protein is an antibody.
-
43. A surface prepared by the method of claim 28.
-
44. A composition prepared by the method of claim 30.
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5. A compound of formula II:
-
B—
R—
A—
NHN═
C(R1R2)
IIor a derivative thereof, wherein;
A is NH(C═
O)—
, NH(C═
S)—
, NHNH(C═
O)—
, or NHNH(C═
S)—
or a direct bond to R;
B is an amino or thiol reactive moiety;
R is an aliphatic divalent group having any combination of the following groups, which are combined in any order;
cycloalkylene, C(R10)2, —
C(R10)═
C(R10)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G)a, P(J)b(R10), P(J)b(LR10), N(R10), >
N+(R12)(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G is O or NR10;
J is S or O; and
L is S, O or NR10;
each R10 is a monovalent group independently selected from hydrogen and M1-R14;
each M1 is a divalent group independently having any combination of the following groups, which groups are combined in any order;
a direct link, arylene, heteroarylene, cycloalkylene, C(R15)2, —
C(R15)═
C(R15)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C, O—
, S(G1)a, P(J)b(R15), P(J)b(LR15), N(R15), N(COR15), >
N+(R12)(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G1 is O or NR15;
J is S or O; and
L is S, O or NR15;
R14 and R15 are each independently selected from the group among hydrogen, halo, pseudohalo, cyano, azido, nitr , SiR16R17R18, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20;
R19 and R20 are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl and heterocyclyl;
R12 and R13 are selected from (i) or (ii) as follows;
(i) R12 and R13 are independently selected from among hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl and heteroaryl;
or (ii) R12 and R13 together form alkylene, alkenylene or cycloalkylene;
R16, R17 and R18 are each independently a monovalent group selected from hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20; and
R11, R12, R13, R14, R15, R16, R17, R18, R19 and R20 can be substituted with one or more substituents each independently selected from Z, wherein Z is selected from alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, hydroxy, S(O)hR30, NR30R31, COOR30, COR30, CONR30R31, OC(O)NR30R31, N(R30)C(O)R31, alkoxy, aryloxy, heteroaryl, heterocyclyl, heteroaryloxy, heterocyclyloxy, aralkyl, aralkenyl, aralkynyl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, aralkoxy, heteroaralkoxy, alkoxycarbonyl, carbamoyl, thiocarbamoyl, alkoxycarbonyl, carboxyaryl, halo, pseudohalo, haloalkyl and carboxamido;
h is 0, 1 or 2; and
R30 and R31 are each independently selected from among hydrogen, halo, pseudohalo, cyano, azido, nitro, trialkylsilyl, dialkylarylsilyl, alkyldiarylsilyl, triarylsilyl, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, h t roaryl, h t roaralkyl, heteroaralkenyl, het roaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, het roaralkoxy, amino, amido, alkylamino, dialkylamino, alkylarylamino, diarylamino and arylamino;
R1 is a saturated straight chain of 3 to 20 carbon atoms, a chain of 2 to 2000 ethyleneoxide moieties, or a saturated or unsaturated carbocyclic moiety of 3 to 20 carbon atoms; and
R2 is a saturated straight chain of 3 to 20 carbon atoms, a chain of 2 to 2000 ethyleneoxide moieties, a saturated or unsaturated carbocyclic moiety of 3 to 20 carbon atoms. - View Dependent Claims (6, 7, 49, 52)
-
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49. The compound of claim 5, wherein B is an amino reactive moiety selected from succininimidyl ester, hydroxybenzotriazolyl ester, or pentafluorophenol ester.
-
52. The compound of claim 5, wherein B is a thiol reactive moiety selected from maleimido, α
- -bromoacetyl or pyridyidisulfide.
-
8. A compound of formula III:
-
B—
R—
(C═
O)—
NHNH2.HX
IIIor a derivative thereof, wherein;
B is an amino reactive moiety;
R is an aliphatic divalent group having any combination of the following groups, which are combined in any order;
cycloalkylene, C(R10)2, —
C(R10)═
C(R10)—
;
>
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G)a, P(J)b(R10), P(J)b(LR10), N(R10), >
N+(R12)(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G is O or NR10;
J is S or O; and
L is S, O or NR10;
each R10 is a monovalent group independently selected from hydrogen and M1-R14;
each M1 is a divalent group independently having any combination of the following groups, which groups are combin d in any order;
a direct link, arylene, het roarylene, cycloalkylene, C(R15)2, —
C(R15)═
C(R15)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C, O, S(G1)a, P(J)b(R15), P(J)b(LR15), N(R15), N(COR15), >
N+(R12)(R13) and C(L);
wh re a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G1 is O or NR15;
J is S or O; and
L is S, O or NR15;
R14 and R15 are each independently selected from the group among hydrogen, halo, pseudohalo, cyano, azido, nitro, SiR16R17R18, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20;
R19 and R20 are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl and heterocyclyl;
R12 and R13 are selected from (i) or (ii) as follows;
(i) R12 and R13 are independently selected from among hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl and heteroaryl;
or (ii) R12 and R13 together form alkylene, alkenylene or cycloalkylene;
R16, R17 and R18 are each independently a monovalent group selected from hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20; and
R11, R12, R13, R14, R15, R16, R17, R18, R19 and R20 can be substituted with one or more substituents each independently selected from Z, wherein Z is selected from alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, hydroxy, S(O)hR30, NR30R31, COOR30, COR30, CONR30R31, OC(O)NR30R31, N(R30)C(O)R31, alkoxy, aryloxy, heteroaryl, heterocyclyl, heteroaryloxy, heterocyclyloxy, aralkyl, aralkenyl, aralkynyl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, aralkoxy, heteroaralkoxy, alkoxycarbonyl, carbamoyl, thiocarbamoyl, alkoxycarbonyl, carboxyaryl, halo, pseudohalo, haloalkyl and carboxamido;
h is 0, 1 or 2; and
R30 and R31 are each independ ntly selected from among hydrogen, halo, ps udohalo, cyano, azido, nitro, trialkylsilyl, dialkylarylsilyl, alkyldiarylsilyl, triarylsilyl, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy, amino, amido, alkylamino, dialkylamino, alkylarylamino, diarylamino and arylamino; and
X is a negative counterion. - View Dependent Claims (9, 45, 50)
-
-
10. A compound that has one of formulae VII or VIII:
-
B—
R—
ONH2.HX
VII;
orB—
R—
ON═
C(R1R2)
VIIIor a derivative thereof, wherein;
R is a divalent group having any combination of the following groups, which are combined in any order;
arylene, heteroarylene, cycloalkylene, C(R10)2, —
C(R10)═
C(R10)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G)a, P(J)b(R10), P(J)b(LR10), N(R10), >
N+(R12)(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G is O or NR10;
J is S or O; and
L is S, O or NR10;
each R10 is a monovalent group independently selected from hydrogen and M1-R4;
each M1 is a divalent group independently having any combination of the following groups, which groups are combined in any order;
a direct link, arylene, heteroarylene, cycloalkylene, C(R15)2, —
C(R15)═
C(R15)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G1)a, P(J)b(R15), P(J)b(LR15), N(R15), N(COR15), >
N+(R12)(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G1 is O or NR15;
J is S or O; and
L is S, O or NR15;
R14 and R15 are each independently selected from the group among hydrogen, halo, pseudohalo, cyano, azido, nitro, SiR16R17R18, alkyl, alk nyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, h teroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalk nyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, h teroaralkoxy and NR19R20;
R19 and R20 are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl and heterocyclyl;
R12 and R13 are selected from (i) or (ii) as follows;
(i) R12 and R13 are independently selected from among hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl and heteroaryl;
or (ii) R12 and R13 together form alkylene, alkenylene or cycloalkylene;
R16, R17 and R18 are each independently a monovalent group selected from hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20; and
R11, R12, R13, R14, R15, R16, R1, R18, R19 and R20 can be substituted with one or more substituents each independently selected from Z, wherein Z is selected from alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, hydroxy, S(O)hR30, NR30R31, COOR30, COR30, CONR30R31, OC(O)NR30R31, N(R30)C(O)R31, alkoxy, aryloxy, heteroaryl, heterocyclyl, heteroaryloxy, heterocyclyloxy, aralkyl, aralkenyl, aralkynyl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, aralkoxy, heteroaralkoxy, alkoxycarbonyl, carbamoyl, thiocarbamoyl, alkoxycarbonyl, carboxyaryl, halo, pseudohalo, haloalkyl and carboxamido;
h is 0, 1 or 2; and
R30 and R31 are each independently selected from among hydrogen, halo, pseudohalo, cyano, azido, nitro, trialkylsilyl, dialkylarylsilyl, alkyldiarylsilyl, triarylsilyl, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy, amino, amido, alkylamino, dialkylamino, alkylarylamino, diarylamino and arylamino;
B is an amino or thiol reactive moiety;
R1 is H or a saturated straight chain of 3 to 20 carbon atoms, a chain of 2 to 2000 ethyleneoxide moieties, a saturated or unsaturated carbocyclic moiety of 3 to 20 carbon atoms;
R2 is a saturated straight chain of 3 to 20 carbon atoms, a chain of 2 to 2000 ethyleneoxide moieties, a saturated or unsaturated carbocyclic moiety of 3 to 20 carbon atoms; and
X is a negative counterion. - View Dependent Claims (11, 12, 13, 46, 51, 53)
-
-
46. The compound of claim 10, wherein X is a halide or trifluoroacetate.
-
51. The compound of claim 10, wherein B is an amino reactive moiety selected from succininimidyl ester, hydroxybenzotriazolyl ester, or pentafluorophenol ester.
-
53. The compound of claim 10, wherein B is a thiol reactive moiety selected from maleimido, α
- -bromoacetyl or pyridyidisulfide.
- 14. A compound that has any of formulae XI:
- 17. A compound that has one of formulae XII:
-
19. A compound that has any of formulae XIII:
-
R3 S—
R—
A—
NHNH2.HX,R30S—
R—
A—
NHN═
CR1R2,(S—
R—
A—
NHNH2.HX)2, or(S—
R—
A—
NHN═
CR1R2)2;or a derivative thereof, wherein R1 is H or a saturated straight chain of 3 to 20 carbon atoms, a chain of 2 to 2000 ethyleneoxide moieties, or a saturated or unsaturated carbocyclic moiety of 3 to 20 carbon atoms;
R2 is a saturated straight chain of 3 to 20 carbon atoms, a chain of 2 to 2000 ethyleneoxide moieties, or a saturated or unsaturated carbocyclic moiety of 3 to 20 carbon atoms;
X is a negative counterion; and
R is a divalent group having any combination of the following groups, which are combined in any order;
arylene, heteroarylene, cycloalkylene, C(R10)2, —
C(R10)═
C(R10)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G)a, P(J)b(R10), P(J)b(LR10), N(R10), >
N+(R12)(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G is O or NR10;
J is S or O; and
L is S, O, or NR10;
each R10 is a monovalent group independently selected from hydrogen and M1-R14;
each M1 is a divalent group independently having any combination of the following groups, which groups are combined in any order;
a direct link, arylene, heteroarylene, cycloalkylene, C(R15)2, —
C(R15)═
C(R15)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G1)a, P(J)b(R15), P(J)b(LR15), N(R15), N(COR15), >
N+(R12(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G1 is O or NR15;
J is S or O; and
L is S, O, or NR15;
R14 and R15 are each independently selected from the group among hydrogen, halo, pseudohalo, cyano, azido, nitro, SiR16R17R18, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, he teroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20;
R19 and R20 are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl and heterocyclyl;
R12 and R13 are selected from (i) or (ii) as follows;
(i) R12 and R13 are independently selected from among hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl and heteroaryl;
or (ii) R12 and R13 together form alkylene, alkenylene or cycloalkylene;
R16, R17 and R18 are each independently a monovalent group selected from hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20; and
R11, R12, R13, R14, R15, R16, R17, R18, R19 and R20 can be substituted with one or more substituents each independently selected from Z, wherein Z is selected from alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, hydroxy, S(O)hR30, NR30R31, COOR30, COR30, CONR30R31, OC(O)NR30R31, N(R30)C(O)R31, alkoxy, aryloxy, heteroaryl, heterocyclyl, heteroaryloxy, heterocyclyloxy, aralkyl, aralkenyl, aralkynyl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, aralkoxy, heteroaralkoxy, alkoxycarbonyl, carbamoyl, thiocarbamoyl, alkoxycarbonyl, carboxyaryl, halo, pseudohalo, haloalkyl and carboxamido;
h is 0, 1 or 2; and
R30 and R31 are each independently selected from among hydrogen, halo, pseudohalo, cyano, azido, nitro, trialkylsilyl, dialkylarylsilyl, alkyldiarylsilyl, triarylsilyl, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy, amino, amido, alkylamino, dialkylamino, alkylarylamino, diarylamino and arylamino;
A is a direct link, C═
O, C═
S, NH(C═
O), NH(C═
S), NHNH(C═
O), or NHNH(C═
S); and
R30 is hydrogen or a thiol protecting group. - View Dependent Claims (20, 47)
-
-
47. The compound of claim 19, wherein X is a halide or trifluoroacetate.
-
21. A compound that has one of formulae XII:
-
R30S—
R—
ONH2.HX,R30S—
R—
ON═
CR1R2,(S—
R—
ONH2.HX)2, or(S—
R—
ON═
CR1R2)2;or a derivative thereof, wher in R1 is H or a saturated straight chain of 3 to 20 carbon atoms, a chain of 2 to 2000 ethyleneoxide moieties, or a saturated or unsaturated carbocyclic moiety of 3 to 20 carbon atoms;
R2 is a saturated straight chain of 3 to 20 carbon atoms, a chain of 2 to 2000 ethyleneoxide moieties, or a saturated or unsaturated carbocyclic moiety of 3 to 20 carbon atoms;
X is a negative counterion; and
R is a divalent group having any combination of the following groups, which are combined in any order;
arylene, heteroarylene, cycloalkylene, C(R10)2, —
C(R10)═
C(R10)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G)a, P(J)b(R10), P(J)b(LR10), N(R10), >
N+(R12)(R13) and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G is O or NR10;
J is S or O; and
L is S, O, or NR10;
each R10 is a monovalent group independently selected from hydrogen and M1-R14;
each M1 is a divalent group independently having any combination of the following groups, which groups are combined in any order;
a direct link, arylene, heteroarylene, cycloalkylene, C(R15)2, —
C(R15)═
C(R15)—
, >
C═
C(R12)(R13), >
C(R12)(R13), —
C≡
C—
, O, S(G1)a, P(J)b(R15), P(J)b(LR15), N(R15), N(COR15), >
N+(R12)(R13), and C(L);
where a is 0, 1 or 2;
b is 0, 1, 2 or 3;
G1 is O or NR15;
J is S or O; and
L is S, O, or NR15;
R14 and R15 are each independently selected from the group among hydrogen, halo, pseudohalo, cyano, azido, nitro, SiR16R17R18, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20;
R19 and R20 are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl and heterocyclyl;
R12 and R13 are selected from (i) or (ii) as follows;
(i) R12 and R13 are independently s lected from among hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl and heteroaryl;
or (ii) R12 and R13 together form alkylene, alkenylene or cycloalkylene;
R16, R17 and R18 are each independently a monovalent group selected from hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy and NR19R20; and
R11, R12, R13, R14, R15, R16, R17, R18, R19 and R20 can be substituted with one or more substituents each independently selected from Z, wherein Z is selected from alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, hydroxy, S(O)hR30, NR30R31, COOR30, COR30, CONR30R31, OC(O)NR30R31, N(R30)C(O)R31, alkoxy, aryloxy, heteroaryl, heterocyclyl, heteroaryloxy, heterocyclyloxy, aralkyl, aralkenyl, aralkynyl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, aralkoxy, heteroaralkoxy, alkoxycarbonyl, carbamoyl, thiocarbamoyl, alkoxycarbonyl, carboxyaryl, halo, pseudohalo, haloalkyl and carboxamido;
h is 0, 1 or 2; and
R30 and R31 are each independently selected from among hydrogen, halo, pseudohalo, cyano, azido, nitro, trialkylsilyl, dialkylarylsilyl, alkyldiarylsilyl, triarylsilyl, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, aryl, aralkyl, aralkenyl, aralkynyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heteroaralkynyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, hydroxy, alkoxy, aryloxy, aralkoxy, heteroaralkoxy, amino, amido, alkylamino, dialkylamino, alkylarylamino, diarylamino and arylamino; and
R30 is hydrogen or a thiol protecting group. - View Dependent Claims (48)
-
-
31. A method of crosslinking a natural or synthetic biological molecule, comprising:
-
(i) preparing a conjugate of formula IVa;
or a derivative thereof, wherein;
A is NH(C═
O), NH(C═
S), NH(C═
NH), NHNH(C═
O), NHNH(C═
S), NHNH(C═
NH) or a direct bond;
B is a natural or synthetic biological molecule;
D is a carbon or nitrogen atom;
E is a carbon or nitrogen atom; and
X is a negative counter ion, oxygen, sulfur or —
NH; and
(ii) applying the conjugate to a surface wherein the surface has at least one carbonyl moiety for a time and under conditions such that the hydrazine moiety of the conjugate reacts with the carbonyl moiety of the surface forming a hydrazone bond to the surface.
-
-
32. A method of crosslinking a natural or synthetic biological molecule, comprising:
-
(i) preparing a conjugate of formula Va;
or a derivative thereof, wherein;
A is NH(C═
O), NH(C═
S), NH(C═
NH), NHNH(C═
O), NHNH(C═
S), NHNH(C═
NH) or a direct bond;
B is a natural or synthetic biological molecule;
D is a carbon or nitrogen atom;
E is a carbon or nitrogen atom;
R1 is hydrogen or a saturated straight chain of 1 to 12 carbon atoms; and
R2 is hydrogen or a saturated straight chain of 1 to 12 carbon atoms; and
(ii) applying the conjugate to a surface wherein the surface has at least one amino or one thiol reactive moiety for a time and under conditions such that the conjugate reacts with the amino or thiol reactive moiety of the surface forming a bond to the surface.
-
-
33. A method of crosslinking a natural or synthetic biological molecule, comprising:
-
(i) preparing a conjugate of the formula VIa;
or a derivative thereof, wherein;
B is a natural or synthetic biological molecule;
D is a carbon or nitrogen atom;
E is a carbon or nitrogen atom; and
X is a negative counter ion, oxygen, sulfur or —
NH; and
(ii) applying the conjugate to a surface wherein the surface has at least one carbonyl moiety for a time and under conditions such that the hydrazine moiety of the conjugate reacts with the carbonyl moiety of the surface forming a hydrazone bond to the surface.
-
-
34. A method of crosslinking a natural or synthetic biological molecule, comprising:
-
(i) preparing a conjugate of formula IVa;
or a derivative thereof, wherein;
A is NH(C═
O), NH(C═
S), NH(C═
NH), NHNH(C═
O), NHNH(C═
S), NHNH(C═
NH) or a direct bond;
B is a natural or synthetic biological molecule;
D is a carbon or nitrogen atom;
E is a carbon or nitrogen atom; and
X is a negative counter ion, oxygen, sulfur or —
NH; and
(ii) applying the conjugate to a surface wherein the surface has at least one amino or one thiol reactive moiety for a time and under conditions such that the conjugate reacts with the amino or thiol reactive moiety of the surface forming a bond to the surface.
-
-
35. A method of crosslinking a natural or synthetic biological molecule, comprising:
-
(i) preparing a conjugate of formula Va;
or a derivative thereof, wherein;
A is NH(C═
O), NH(C═
S), NH(C═
NH), NHNH(C═
O), NHNH(C═
S), NHNH(C═
NH) or a direct bond;
B is a natural or synthetic biological molecule;
D is a carbon or nitrog n atom;
E is a carbon or nitrogen atom;
R1 is hydrogen or a saturated straight chain of 1 to 12 carbon atoms; and
R2 is hydrogen or a saturated straight chain of 1 to 12 carbon atoms; and
(ii) applying the conjugate to a surface wherein the surface has at least one amino or one thiol reactive moiety for a time and under conditions such that the conjugate reacts with the amino or thiol reactive moiety of the surface forming a bond to the surface.
-
-
36. A method of crosslinking a natural or synthetic biological molecule, comprising:
-
(i) preparing a conjugate of formula VIa;
or a derivative thereof, wherein;
B is a natural or synthetic biological molecule;
D is a carbon or nitrogen atom;
E is a carbon or nitrogen atom; and
X is a negative counter ion, oxygen, sulfur or —
NH; and
(ii) applying the conjugate to a surface wherein the surface has at least one amino or one thiol reactive moiety for a time and under conditions such that the conjugate reacts with the amino or thiol reactive moiety of the surface forming a bond to the surface.
-
-
37. A method of crosslinking a natural or synthetic biological molecule, comprising:
-
(i) preparing a conjugate of formula IVa;
or a derivative thereof, wherein;
A is NH(C═
O), NH(C═
S), NH(C═
NH), NHNH(C═
O), NHNH(C═
S), NHNH(C═
NH) or a direct bond;
B is a natural or synthetic biological molecule;
D is a carbon or nitrogen atom;
E is a carbon or nitrogen atom; and
X is a negative counter ion, oxygen, sulfur or —
NH; and
(ii) mixing the conjugate to a natural or synthetic biological molecule, wherein the molecule has at least one carbonyl moiety, for a time and under conditions such that the hydrazine moiety of the conjugate reacts with the carbonyl moiety of the molecule forming a hydrazone bond to the molecule.
-
-
38. A method of crosslinking a natural or synthetic biological molecule, comprising:
-
(i) preparing a conjugate of formula Va;
or a derivative thereof, wherein;
A is NH(C═
O), NH(C═
S), NH(C═
NH), NHNH(C═
O), NHNH(C═
S), NHNH(C═
NH) or a direct bond;
B is a natural or synthetic biological molecule;
D is a carbon or nitrogen atom;
E is a carbon or nitrogen atom;
R1 is hydrogen or a saturated straight chain of 1 to 12 carbon atoms; and
R2 is hydrogen or a saturated straight chain of 1 to 12 carbon atoms; and
(ii) mixing the conjugate with a natural or synthetic biological molecule, wherein the molecule has at least one carbonyl moiety, for a time and under conditions such that the hydrazine moiety of the conjugate reacts with the carbonyl moiety of the molecule forming a hydrazone bond to the molecule.
-
-
39. A method of crosslinking a natural or synthetic biological molecule, comprising:
-
(i) preparing a conjugate of formula VIa;
or a derivative thereof, wherein;
B is a natural or synthetic biological molecule;
D is a carbon or nitrogen atom;
E is a carbon or nitrogen atom; and
X is a negative counter ion, oxygen, sulfur or —
NH; and
(ii) mixing the conjugate with a natural or synthetic biological molecule, wherein the molecule has at least one carbonyl moiety for a time and under conditions such that the hydrazine moiety of the conjugate reacts with the carbonyl moiety of the molecule forming a hydrazone bond to the molecule.
-
Specification