Hydrazinyl-substituted heteroaryl compounds and methods for producing a conjugate

1. A compound of formula (II):

wherein:

R 8 is H and R 9 is (CR 2 R 4 )(NR 6 )(NHR 5 ), or R 8 is (CR 1 R 3 )(NR 5 )(NHR 6 ) and R 9 is H;

R 1 , R 2 , R 3 and R 4 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;

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

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

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

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

6. The compound of claim 1 , wherein R 7 is hydrogen.

7. The compound 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) n , (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 are 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.

8. The compound of claim 7 , 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.

9. The compound of claim 8 , 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.

10. The compound of claim 8 , 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 any two adjacent R 12 groups are cyclically linked to form a piperazinyl ring.

11. The compound of claim 8 , wherein each R 13 is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl.

12. The compound of claim 8 , 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.

13. The compound of claim 8 , 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—

(C 1 -C 12 )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

—

—

—.

14. The compound of claim 8 , 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.

15. A method of producing a polypeptide conjugate, the method comprising:

combining in a reaction mixture:

a compound of claim 1 , and

a polypeptide comprising a reactive aldehyde group or a reactive ketone group,

wherein the combining is under reaction conditions suitable to promote reaction between the compound and the reactive aldehyde group or reactive ketone group of the polypeptide to form a polypeptide conjugate; and

isolating the polypeptide conjugate from the reaction mixture.