Conjugates of Pyrrolo[1,4]Benzodiazepine Dimers As Anticancer Agents

Abstract

The present invention relates to pyrrolo[1,4]benzodiazepine (PBD) dimer conjugates, to the compositions comprising them and to their therapeutic application, in particular as anticancer agents. The invention also relates to the process for the preparation of the conjugates, to their application as anticancer agents and to the dimers themselves.

1 .- 9 . (canceled) 
     
     
         10 . A process for the preparation of a conjugate, said process comprising:
 (i) bringing into contact and allowing to react an optionally buffered aqueous solution of a binding agent and a solution of a compound according to formula (I):   
       
         
           
           
               
               
           
         
         wherein:
    represents a single bond or a double bond; 
 R 1  and R 2 , which are identical or different, represent, independently of one another: H, Hal or a (C 1 -C 6 )alkyl group optionally substituted by one or more substituent(s) chosen from: Hal, CN, NRR′, CF 3 , OR, an aryl or heteroaryl group, or S(O) q R with q=0, 1 or 2; or R 1  and R 2  together form a double bond ═CH 2  or ═CH—CH 3 ; 
 R 1 ′ and R 2 ′, which are identical or different, represent, independently of one another: H, Hal or a (C 1 -C 6 )alkyl group optionally substituted by one or more substituent(s) chosen from: Hal, CN, NRR′, CF 3 , OR, an aryl or heteroaryl group, or S(O) q R with q=0, 1 or 2; or R 1 ′ and R 2 ′ together form a double bond ═CH 2  or ═CH—CH 3 ; 
 Y and Y′, which are identical or different, represent, independently of one another, H or OR; 
 M represents CH or N; 
 ALK and ALK′, which are identical or different, represent, independently of one another, a (C 1 -C 6 )alkylene group; 
 R and R′ represent, independently of one another, H or a (C 1 -C 6 )alkyl or aryl group optionally substituted by one or more substituent(s) chosen from: Hal, CN, NRR′, CF 3 , or an aryl or heteroaryl group; 
 L represents an -L 1 -L 2 - group, wherein:
 L 1  is attached to the aromatic ring comprising M via the ALK or OALK group and represents one of the following groups:
 -ALK-S—; 
 
 
 
       
       
         
           
           
               
               
           
         
         
           
             
               —O-ALK-NR 3 -ALK-S—; 
             
           
         
       
       
         
           
           
               
               
           
         
         
           
             and L 2  represents the —CH 2 C(═O)—NR 3 —(CH 2 CH 2 O) i -ALK- group attached to L 1  via —CH 2 C(═O)—; 
           
           or L represents the —O-ALK-NR 3 -ALK-S—(CH 2 CH 2 O) i -ALK- group attached to the aromatic ring comprising M via the OALK group; 
           R 3  represents H or a (C 1 -C 6 )alkyl group; 
           i represents an integer ranging from 1 to 40, rather from 1 to 20, preferably from 1 to 10; and 
           Z b  represents a single bond, —O— or —NH— and R b  represents H or a (C 1 -C 6 )alkyl, (C 3 -C 7 )cycloalkyl, aryl, heteroaryl or (C 3 -C 7 )heterocycloalkyl group; 
           or Z b  represents a single bond and R b  represents Hal; 
         
         provided that when   represents a single bond, then:
    represents a single bond; 
 U and U′ each is H; and 
 W and W′, which are identical or different, each represents, independently of one another, —OH, —OR, —OCOR, —COOR, —OCOOR, —OCONRR′, a cyclic carbamate such that N10 and C11 are included in a ring, —NRCONRR′, —OCSNHR, a cyclic thiocarbamate such that N10 and C11 are included in a ring, —SH, —SR, —SOR, —SOOR, —SO 3   − , —NRSOOR′, —NRR′, a cyclic amine such that N10 and C11 are included in a ring, —NROR′, —NRCOR′, —N 3 , —CN, Hal or a trialkylphosphonium or triarylphosphonium group; 
 
         and 
         (ii) optionally separating the conjugate formed in stage (i) from the compound of formula (I) and/or from the binding agent not having reacted and/or from the aggregates which might be formed. 
       
     
     
         11 . The process according to  claim 10 , in which the —C(═O)Z b R b  group of said compound of formula (I) is reactive with regard to the chemical groups present on the binding agent, in particular with regard to the amino groups present on an antibody, so as to ensure the attachment of the compound of formula (I) to the binding agent by formation of a covalent bond. 
     
     
         12 . The process according to  claim 10 , in which step (ii) comprises:
 separating the conjugate which has formed in stage (i) from the unreacted binding agent and from the aggregates possibly present in the solution;   or   separating the conjugate of stage (i) only from the unreacted compound of formula (I) and from the aggregates which might be formed and leaving in the solution the binding agent which might not have reacted.   
     
     
         13 . The process according to  claim 10 , in which the binding agent is a ligand, a protein, an antibody, a protein or antibody fragment, a peptide, an oligonucleotide or an oligosaccharide. 
     
     
         14 . The process according to  claim 10 , in which the reaction takes place at a temperature of between 20 and 40° C. and/or the duration of the reaction varies between 1 and 24 h. 
     
     
         15 . The process according to  claim 10 , in which, after stage (i) or (ii), the solution of the conjugate is subjected to a stage (iii) of ultrafiltration and/or of diafiltration. 
     
     
         16 .- 38 . (canceled) 
     
     
         39 . The process according to  claim 10 , wherein the compound of formula (I) is according to formula: 
       
         
           
           
               
               
           
         
       
     
     
         40 . The process according to  claim 10 , wherein the compound of formula (I) is according to formula (IA) or (IB): 
       
         
           
           
               
               
           
         
       
     
     
         41 . The process according to  claim 10 , wherein each of Y and Y′ represent, independently, a (C 1 -C 4 )alkoxy group. 
     
     
         42 . The process according to  claim 41 , wherein each of Y and Y′ represent a methoxy group. 
     
     
         43 . The process according to  claim 10 , wherein L is:
 -ALK-S—CH 2 C(═O)—NH—(CH 2 CH 2 O) i —CH 2 CH 2 —;   
       
         
           
           
               
               
           
         
         —O-ALK-NR 3 -ALK-S—CH 2 C(═O)—NH—(CH 2 CH 2 O) i —CH 2 CH 2 ; 
       
       
         
           
           
               
               
           
         
       
       or
 —O-ALK-NR 3 -ALK-S—(CH 2 CH 2 O) i —CH 2 CH 2 . 
 
     
     
         44 . The process according to  claim 10 , wherein L is: 
       
         
           
           
               
               
           
         
       
     
     
         45 . The process according to  claim 10 , wherein L is: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         46 . The process according to  claim 10 , wherein —COZ b R b  represents:
 —COOH; 
 —COO(C 1 -C 6 )alkyl, —COOCH 3 , —COOCH 2 CH═CH 2 ; 
 
       
         
           
           
               
               
           
         
       
       in which IG represents one or more inductive group, 
     
     
         47 . The process according to  claim 46 , in which IG represents: 
       
         
           
           
               
               
           
         
       
     
     
         48 . The process according to  claim 10 , wherein the compound of formula (I)
 is