Hydrazine-based and carbonyl-based bifunctional crosslinking reagents
First Claim
Patent Images
1. 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(L1R15), N(R5), N(COR15), >
N+(R12)(R13) and C(L1);
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
L1 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
R10, 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;
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, or a saturated or unsaturated carbocyclic moiety of 3 to 20 carbon atoms.
18 Assignments
0 Petitions
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
7 Claims
-
1. 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(L1R15), N(R5), N(COR15), >
N+(R12)(R13) and C(L1);
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
L1 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; andR10, 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;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, or a saturated or unsaturated carbocyclic moiety of 3 to 20 carbon atoms. - View Dependent Claims (6, 7)
-
-
2. 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 a saturated straight chain of 1 to 20 carbon atoms, a chain of 2 to 2000 ethyleneoxide moieties, a saturated or an unsaturated carbocyclic moiety of 3 to 20 carbon atoms or 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(R2)(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(L1R15), N(R15), N(COR15), >
N+(R12)(R13) and C(L1);
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
L1 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; andR10, 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)R30, alkoxy, aryloxy, heteroaryl, heterocyclyl, heteroaryloxyn, heterocyclylox, 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, dialkylarmlsilyl, 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;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, or a saturated or unsaturated carbocyclic moiety of 3 to 20 carbon atoms.
-
-
3. A compound of the formula:
-
4. A method of crosslinking a natural or synthetic biological molecule, comprising:
(i) preparing a conjugate of formula Va;
-
5. A method of crosslinking a natural or synthetic biological molecule, comprising:
(i) preparing a conjugate of formula Va;
Specification