Hydrazinyl-pyrrolo compounds and methods for producing a conjugate

1. A compound of formula (VIIIa):

wherein,

R 2 and R 3 are each alkyl;

Y 1 , Y 2 , Y 3 , and Y 4 are each hydrogen;

L is a linker represented by -(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 and c are each 1, and d and e are each independently 0 or 1, where the sum of a, b, c, d and e is 3 to 5;

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

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CONR 11 —

(PEG) n

—CO—

(EDA) w

C 12 )alkylene

(C 1 -

—CO—

(EDA) w

—CO—

(CR 13 OH) h

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(C 1 -

C 12 )alkylene

C 12 )alkylene

(C 1 -

—CONR 11 —

(PEG) n

—CO—

(AA) p

C 12 )alkylene

(C 1 -

—CONR 11 —

(PEG) n

—CO—

(AA) p

C 12 )alkylene

(C 1 -

—CONR 11 —

(PEG) n

—CO—

(AA) p

C 12 )alkylene

(C 1 -

—CONR 11 —

(PEG) n

—CO—

(AA) p

C 12 )alkylene

(C 1 -

—CO—

(EDA) w

—CO—

(CR 13 OH) h

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(C 1 -

C 12 )alkylene

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(C 1 -

C 12 )alkylene

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CONR 11 —

(PEG) n

—CO—

(AA) p

C 12 )alkylene

(C 1 -

—CONR 11 —

(PEG) n

—CO—

(AA) p

C 12 )alkylene

(C 1 -

—CO—

(EDA) w

—CO—

(CR 13 OH) h

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CONR 11 —

(PEG) n

—CO—

MABO

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CO—

(AA) p

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

—CONR 11 —

(PEG) n

—CO—

MABC

C 12 )alkylene

(C 1 -

—CONR 11 —

substituted

—NR 11 —

(PEG) n

C 12 )alkylene

(C 1 -

C 12 )alkylene

(C 1 -

—CONR 11  

(PEG) n

—CO—

(AA) p

C 12 )alkylene

(C 1 -

—CONR 11  

(C 1 -

—

(CR 13 OH) h

C 12 )alkylene

C 12 )alkylene

(C 1 -

—CO—

P4A

—CO—

(C 1 -

C 12 )alkylene

C 12 )alkylene

(C 1 -

—CO—

P4A

—CO—

(C 1 -

C 12 )alkylene

C 12 )alkylene

(C 1 -

—CO—

P4A

—CO—

(C 1 -

C 12 )alkylene

C 12 )alkylene

(C 1 -

—CO—

P4A

—CO—

(C 1 -

C 12 )alkylene

C 12 )alkylene

V 3

T 4

V 4

T 5

V 5

 CO 

 NR 11  

 CONR 11  

(C 1 -

 CO 

C 12 )alkylene

 CO 

MABO

PABO

PABC

 CO 

(AA) p

 CO 

(AA) p

 CO 

(AA) p

 CO 

(AA) p

 CO 

(AA) p -

PABC-

(AA) p

 CO 

(AA) p

PABC-

(AA) p

 CO 

(AA) p

PABO

 CO 

(AA) p

PABO

 SO 2  

(AA) p

PABC-

(AA) p

PABC

(AA) p

 CONR 11  

(PEG) n

 CO 

 CO 

MABC-

(AA) p

 CO 

MABC

(AA) p

 CO 

MABO

 CO 

PABO

 CO 

PABC

(AA) p

 CO 

PABC

 NR 11  

 CONR 11  

 CO 

(AA) p

PABO

 CO 

 CO 

(AA) p

PABO

 CO 

(AA) p

PABC-

(AA) p

 CO 

(AA )p

wherein EDA is an ethylene diamine moiety, PEG is a polyethylene glycol, and AA is an amino acid residue;

w is an integer from 1 to 20;

n is an integer from 1 to 30;

p is an integer from 1 to 20;

h is an integer from 1 to 12;

R 13 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;

R 11 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; and,

W 1 is a drug.

2. The compound of claim 1 ,

wherein:

(PEG) n is

 where n is an integer from 1 to 30;

EDA is an ethylene diamine moiety having the following structure:

 where q is an integer from 1 to 6 and r is 0 or 1;

piperidin-4-amino (P4A) is

each R 11 and R 12 is independently selected from hydrogen, an alkyl, a substituted alkyl, a polyethylene glycol moiety, an aryl and a substituted aryl, wherein any two adjacent R 12 groups may be cyclically linked to form a piperazinyl ring; and

R 13 is selected from hydrogen, an alkyl, a substituted alkyl, an aryl, and a substituted aryl.

3. The compound of claim 1 , wherein L is one of the following structures:

wherein:

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

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

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

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

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

each R is independently hydrogen, alkyl, substituted alkyl, a polyethylene glycol moiety, 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 H, 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.

4. The compound of claim 1 , wherein the drug comprises a maytansinoid.

5. The compound of claim 4 , wherein the maytansinoid is deacyl maytansine.

6. The compound of claim 1 , wherein the compound is of the formula:

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

combining in a reaction mixture:

a compound of claim 1 , and

a second compound 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 second compound to form the conjugate; and

isolating the conjugate from the reaction mixture.

8. The method of claim 7 , wherein the second compound comprises a polypeptide.

9. The method of claim 7 , wherein the compound is of the formula:

10. A pharmaceutical composition comprising:

a compound of claim 1 ; and

a pharmaceutically acceptable excipient.

11. The pharmaceutical composition of claim 10 , wherein the compound is of the formula:

12. A method of treating cancer by administering to a patient having said cancer a therapeutically effective amount of the compound of claim 1 .

13. The method of claim 12 , wherein the cancer is carcinoma, sarcoma, leukemia, or lymphoma.

14. The method of claim 13 , wherein the carcinoma is a carcinoma selected from the group consisting of esophageal carcinoma, hepatocellular carcinoma, basal cell carcinoma, squamous cell carcinoma, bladder carcinoma, bronchogenic carcinoma, colon carcinoma, colorectal carcinoma, gastric carcinoma, lung carcinoma, adrenocortical carcinoma, thyroid carcinoma, pancreatic carcinoma, breast carcinoma, ovarian carcinoma, prostate carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, renal cell carcinoma, ductal carcinoma in situ or bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical carcinoma, uterine carcinoma, testicular carcinoma, osteogenic carcinoma, epithelial carcinoma, and nasopharyngeal carcinoma.

15. The method of claim 13 , wherein the sarcoma is a sarcoma selected from the group consisting of fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, chordoma, osteogenic sarcoma, osteosarcoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's sarcoma, leiomyosarcoma, and rhabdomyosarcoma.

16. The method of claim 13 , wherein the leukemia is a leukemia selected from the group consisting of chronic myeloproliferative syndromes, acute myelogenous leukemias, chronic lymphocytic leukemias, and acute lymphoblastic leukemias.

17. The method of claim 13 , wherein the lymphoma is a lymphoma selected from the group consisting of B-cell lymphomas, Hodgkin's lymphoma, and non-Hodgkin's B cell lymphoma.

18. The method of claim 12 , wherein the cancer is multiple myeloma.

19. The method of claim 12 , wherein the cancer is a solid tumor.

20. The method of claim 19 , wherein the solid tumor is selected from the group consisting of glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma, and retinoblastoma.