Hydrazinyl-substituted heteroaryl compounds and methods for producing a conjugate

1. A conjugate comprising at least one modified amino acid residue of formula (I):

wherein:

R 1 and R 2 are each independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl;

one of R 3 and R 4 is a polypeptide and the other is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl;

R 5 and R 6 are each independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, or R 5 and R 6 are cyclically linked to form a 5 or 6-membered heterocyclyl;

Z 1 is selected from CR 7 , N, O and S;

Z 2 is C or N;

R 7 is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl;

L is a linker; and

W is a drug or a detectable label.

2. The conjugate of claim 1 , wherein Z 1 and Z 2 are each N.

3. The conjugate of claim 1 , wherein Z 1 is CR 7 and Z 2 is N.

4. The conjugate of claim 1 , wherein Z 1 is O and Z 2 is C.

5. The conjugate of claim 1 , wherein R 3 is the polypeptide.

6. The conjugate of claim 1 , wherein R 4 is the polypeptide.

7. The conjugate of claim 1 , wherein R 5 and R 6 are each independently selected from alkyl and substituted alkyl.

8. The conjugate of claim 1 , wherein R 7 is hydrogen.

9. The conjugate of claim 1 , wherein:

the linker is of the formula -(T 1 -V 1 ) a -(T 2 -V 2 ) b -(T 3 -V 3 ) c -(T 4 -V 4 ) d -(T 5 -V 5 ) e —, wherein a, b, c, d and e are each independently 0 or 1, where the sum of a, b, c, d and e is 1 to 5;

T 1 , T 2 , T 3 , T 4 and T 5 are each independently selected from (C 1 -C 12 )alkyl, substituted (C 1 -C 12 )alkyl, (EDA) w , (PEG), (AA) p , —(CR 13 OH) h —, 4-amino-piperidinyl (4AP), para-aminobenzyl (PAB), para-amino-benzyloxy (PABO), meta-amino-benzyloxy (MABO), para-amino-benzyloxycarbonyl (PABC), meta-amino-benzyloxycarbonyl (MABC), an acetal group, a disulfide, a hydrazine, a carbohydrate, a beta-lactam, an ester, (AA) p -MABO, (AA) p -MABC, (AA) p -PABO, (AA) p -PABC, MABO-(AA) p , MABC-(AA) p , PABO-(AA) p , PABC-(AA) p , (AA) p -MABO-(AA) p , (AA) p -MABC-(AA) p , (AA) p -PABO-(AA) p , and (AA) p -PABC-(AA) p ;

V 1 , V 2 , V 3 , V 4 and V 5 are each independently selected from the group consisting of a covalent bond, —CO—, —NR 11 —, —CONR 11 —, —NR 11 CO—, —C(O)O—, —OC(O)—, —O—, —S—, —S(O)—, —SO 2 —, —SO 2 NR 11 —, —NR 11 SO 2 — and —P(O)OH—;

each R 11 and R 13 is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl;

w is an integer from 1 to 20;

n is an integer from 1 to 30;

p is an integer from 1 to 20; and

h is an integer from 1 to 12.

10. The conjugate of claim 9 , wherein:

EDA is an ethylene diamine having the structure:

 wherein q is an integer from 1 to 6, r is 0 or 1, and each R 12 is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, or wherein two adjacent R 12 groups are cyclically linked to form a piperazinyl ring;

PEG is a polyethylene glycol or a substituted polyethylene glycol;

AA is an amino acid residue; and

4AP is

 wherein each R 14 is independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol, substituted polyethylene glycol, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

11. The conjugate of claim 10 , wherein each R 11 is independently selected from hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

12. The conjugate of claim 10 , wherein each R 12 is independently selected from hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, or wherein two adjacent R 12 groups are cyclically linked to form a piperazinyl ring.

13. The conjugate of claim 10 , wherein each R 13 is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl.

14. The conjugate of claim 10 , wherein each R 14 is independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol, substituted polyethylene glycol, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

15. The conjugate of claim 10 , wherein T 1 , T 2 , T 3 , T 4 and T 5 and V 1 , V 2 , V 3 , V 4 and V 5 are selected from the following table:

T 1

V 1

T 2

V 2

T 3

V 3

T 4

V 4

T 5

V 5

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

—

—

—

—

(C 1 -C 12 )alkyl

—CO—

(EDA) w

—CO—

(CR 13 OH) h

—CONR 11 —

(C 1 -C 12 )alkyl

—CO—

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(C 1 -C 12 )alkyl

—CO—

—

—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

(PEG) n

—CO—

—

—

—

—

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—

—

—

—

—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

(PEG) n

—NR 11 —

—

—

—

—

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—NR 11 —

—

—

—

—

(C 1 -C 12 )alkyl

—CO—

(EDA) w

—CO—

—

—

—

—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

(C 1 -C 12 )alkyl

—NR 11 —

—

—

—

—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

(PEG) n

—CO—

(EDA) w

—

—

—

—

—

(C 1 -C 12 )alkyl

—CO—

(EDA) w

—

—

—

—

—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

(PEG) n

—CO—

(AA) p

—

—

—

—

—

(C 1 -C 12 )alkyl

—CO—

(EDA) w

—CO—

(CR 13 OH) h

—CO—

(AA) p

—

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(C 1 -C 12 )alkyl

—CO—

(AA) p

—

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

(AA) p

—

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—SO 2 —

(AA) p

—

—

—

(C 1 -C 12 )alkyl

—CO—

(CR 13 OH) h

—CO—

—

—

—

—

—

—

(C 1 -C 12 )alkyl

—CO—

(EDA) w

—CO—

(CR 13 OH) h

—CONR 11 —

(PEG) n

—CO—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

substituted

—NR 11 —

(PEG) n

—CO—

—

—

—

—

(C 1 -C 12 )alkyl

(C 1 -C 12 )alkyl

—SO2—

(C 1 C 12 )alkyl

—CO—

—

—

—

—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

(C 1 -C 12 )alkyl

—

(CR 13 OH) h

—CONR 11 —

—

—

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

(AA) p

—NR 11 —

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—P(O)OH—

(AA) p

—

—

—

(C 1 -C 12 )alkyl

—CO—

(EDA) w

—CO—

(AA) p

—

—

—

—

—

(C 1 -C 12 )alkyl

—CO—

(EDA) w

—CO—

(CR 13 OH) h

—CONR 11 —

(C 1 -C 12 )alkyl

—CO—

(AA) p

—

(C 1 -C 12 )alkyl

—CONR 11 —

(C 1 -C 12 )alkyl

—NR 11 —

—

—CO—

—

—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

(C 1 -C 12 )alkyl

—NR 11 —

—

—CO—

(C1-C1 2 )alkyl

—NR 11 —

—

—

(C 1 -C 12 )alkyl

—CO—

(EDA) w

—CO—

(CR 13 OH) h

—CONR 11 —

(PEG) n

—CO—

(AA) p

—

(C 1 -C 12 )alkyl

—CO—

4AP

—CO—

(C 1 -C 12 )alkyl

—CO—

—

—

—

—

(C 1 -C 12 )alkyl

—CO—

4AP

—CO—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

MABO

—

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

MABC

—

(AA) p

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

(AA) p -

—NR 11 —

(C 1 -C 12 )alkyl

—CO—

PABC

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

(AA) p

—

PABC

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

(AA) p

—

PABO

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

(AA) p -

—

—

—

PABC-(AA) p

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

(AA) p

—

PABC-(AA) p

—

(C 1 -C 12 )alkyl

—CONR 11 —

(PEG) n

—CO—

(AA) p

—

PABO

—

—

—

(C 1 -C 12 )alkyl

—CO—

4AP

—CO—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—

PABO

—

(C 1 -C 12 )alkyl

—CONR 11 —

(PEG) n

—CO—

(AA) p

—

MABO

—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

(PEG) n

—CO—

(AA) p

—

PABC

—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

(PEG) n

—CO—

(AA) p

—

MABC

—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

(PEG) n

—CO—

(AA) p

—

PABC

—

(AA) p

—

(C 1 -C 12 )alkyl

—CONR 11 —

(PEG) n

—CO—

MABO

—

—

—

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

PABO

—

—

—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—NR 11 —

(PEG) n

—CO—

PABC

—

—

—

(C 1 -C 12 )alkyl

—CONR 11 —

(PEG) n

—CO—

MABC

—

(AA) p

—

—

—

(C 1 -C 12 ))alkyl

—CONR 11 —

(PEG) n

—CO—

(AA) p

—

PABC

—NR 11 —

—

—

(C 1 -C 12 )alkyl

—CO—

4AP

—CO—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—

PABC

—

(C 1 -C 12 )alkyl

—CO—

4AP

—CO—

(C 1 -C 12 )alkyl

—CO—

(AA)p

—

PABC-(AA) p

—

(C 1 -C 12 )alkyl

—CO—

4AP

—CO—

(C 1 -C 12 )alkyl

—CO—

(AA) p

—

—

 —.

16. The conjugate of claim 10 , wherein the linker is selected from one of the following structures:

wherein:

each f is independently 0 or an integer from 1 to 12;

each n is independently 0 or an integer from 1 to 30;

each y is independently 0 or an integer from 1 to 20;

each h is independently 0 or an integer from 1 to 12;

each p is independently 0 or an integer from 1 to 20;

each R is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; and

each R′ is independently selected from hydrogen, a sidechain group of an amino acid, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

17. A pharmaceutical composition comprising:

a conjugate of claim 1 ; and

a pharmaceutically acceptable excipient.