Novel cryptophycin compounds and conjugates, their preparation and their therapeutic use

Abstract

The present invention relates to cryptophycin compounds of formula (I):The invention also relates to cryptophycin payloads, to cryptophycin conjugates, to compositions containing them and to their therapeutic use, especially as anticancer agents. The invention also relates to the process for preparing these conjugates.

1 . A Cryptophycin compound of formula (I): 
       
         
           
           
               
               
           
         
         wherein: 
         R 1  represents a (C 1 -C 6 )alkyl group; 
         R 2  and R 3  represent, independently of each other, a hydrogen atom or a (C 1 -C 6 )alkyl group, or alternatively R 2  and R 3  form together with the carbon atom to which they are attached a (C 3 -C 6 )cycloalkyl or a (C 3 -C 6 )heterocycloalkyl group; 
         R 4  and R 5  represent, independently of each other, a hydrogen atom or a (C 1 -C 6 )alkyl group or a (C 1 -C 6 )alkyl-NH(R 2 ) group or a (C 1 -C 6 )alkyl-OH group or a (C 1 -C 6 )alkyl-SH group or a (C 1 -C 6 )alkyl-CO 2 H group, or alternatively R 4  and R 5  form together with the carbon atom to which they are attached a (C 3 -C 6 )cycloalkyl or a (C 3 -C 6 )heterocycloalkyl group; 
         R 6  represents a hydrogen atom or a (C 1 -C 6 )alkyl group; 
         R 7  and R 8  represent, independently of each other, a hydrogen atom or a (C 1 -C 6 )alkyl group or a (C 1 -C 6 )alkyl-CO 2 H group or a (C 1 -C 6 )alkyl-N(C 1 -C 6 )alkyl 2  group or alternatively R 7  and R 8  form together with the carbon atom to which they are attached a (C 3 -C 6 )cycloalkyl group or a (C 3 -C 6 )heterocycloalkyl group; 
         R 9  represents one or more substituents of the phenyl nucleus chosen, independently of each other, from: a hydrogen atom, —OH, (C 1 -C 4 )alkoxy, a halogen atom, —NH 2 , —NH(C 1 -C 6 )alkyl or —N(C 1 -C 6 )alkyl 2  or —NH(C 1 -C 6 )cycloalkyl or (C 3 -C 6 )heterocycloalkyl; 
         R 10  represents at least one substituent of the phenyl nucleus chosen from a hydrogen atom and (C 1 -C 4 )alkyl; 
         W represents
 (C 1 -C 6 )alkyl-NH(R 11 ), 
 (C 1 -C 6 )alkyl-OH, 
 (C 1 -C 6 )alkyl-SH, 
 CO 2 H or C(═O)NH 2 , 
 (C 1 -C 6 )alkyl-CO 2 H or (C 1 -C 6 )alkyl-C(═O)NH 2 , or 
 (C 1 -C 6 )alkyl-N 3 ; and 
 
         R 11  and R 12  represent, independently of each other, a hydrogen atom or a (C 1 -C 6 )alkyl group. 
       
     
     
         2 - 3 . (canceled) 
     
     
         4 . The Cryptophycin compound of formula (I) according to  claim 1 , wherein:
 each of R 2  and R 3  represents a hydrogen atom, or   one of R 2  and R 3  represents a methyl group and the other one represents a hydrogen atom, or R 2  and R 3  form together with the carbon atom to which they are attached a cyclopropyl group.   
     
     
         5 - 6 . (canceled) 
     
     
         7 . The Cryptophycin compound of formula (I) according to  claim 1 , wherein each of R 4  and R 5  represents a methyl group. 
     
     
         8 . (canceled) 
     
     
         9 . The Cryptophycin compound of formula (I) according to  claim 1 , wherein R 7  and R 8  represent, independently of each other, a hydrogen atom or a (C 1 -C 6 )alkyl group. 
     
     
         10 . The Cryptophycin compound of formula (I) according to  claim 1 , wherein R 9  represents two substituents selected from a methoxy group and a chlorine atom. 
     
     
         11 - 13 . (canceled) 
     
     
         14 . The Cryptophycin compound of formula (I) according to  claim 1 , wherein W is selected from (C 1 -C 6 )alkyl-NHR 11 , (C 1 -C 6 )alkyl-OH, (C 1 -C 6 )alkyl-SH and (C 1 -C 6 )alkyl-CO 2 H. 
     
     
         15 . The Cryptophycin compound of formula (I) according to  claim 1 , wherein W represents a CH 2 —NH 2  or a CH 2 —OH group. 
     
     
         16 . (canceled) 
     
     
         17 . A Cryptophycin payload of formula (II): 
       
         
           
           
               
               
           
         
         wherein 
         R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9  and R 10  are as defined in  claim 1 ; 
         Y is selected from (C 1 -C 6 )alkyl-NR 11 , (C 1 -C 6 )alkyl-O, (C 1 -C 6 )alkyl-S, C(═O)O, C(═O)NH, (C 1 -C 6 )alkyl-C(═O)O, (C 1 -C 6 )alkyl-C(═O)NH or (C 1 -C 6 )alkyl-triazole group; 
         R 1  represents a hydrogen atom or a (C 1 -C 6 )alkyl group; 
         L represents a linker; and 
         RCG 1  represents a reactive chemical group present at the end of the linker. 
       
     
     
         18 . (canceled) 
     
     
         19 . The Cryptophycin payload of formula (II) according to  claim 17 , wherein Y represents (C 1 -C 6 )alkyl-NR 11 . 
     
     
         20 . (canceled) 
     
     
         21 . The Cryptophycin payload of formula (II) according to  claim 17 , wherein L is of formula (IV): 
       
         
           
           
               
               
           
         
         wherein:
 L1 represents
 a single bond or a NR 16 (hetero)aryl-CR 15 R 14 —O—C(═O) group if Y═(C 1 -C 6 )alkyl-N(R 11 ), or L1 represents 
 a NR 18 —(C 2 -C 6 )alkyl-NR 17 —C(═O) group or a NR 16 (hetero)aryl-CR 15 R 14 —O—C(═O)—NR 18 —(C 2 -C 6 )alkyl-NR 17 —C(═O) group if Y is (C 1 -C 6 )alkyl-O or (C 1 -C 6 )alkyl-S, or L1 represents 
 a NR 16 (hetero)aryl-CR 15 R 14 — group if Y is C(═O)O, C(═O)NH, (C 1 -C 6 )alkyl-C(═O)O or (C 1 -C 6 )alkyl-C(═O)NH; 
 
 R 11 , R 14 , R 15 , R 16 , R 17  and R 18  each represent, independently of each other, a hydrogen atom or a (C 1 -C 6 )alkyl group; 
 (AA) w  represents a sequence of w amino acids AA connected together via peptide bonds; 
 w represents an integer ranging from 1 to 12; 
 L2 represents a single bond, a (C 1 -C 6 )alkyl group, a (C 1 -C 6 )alkyl-(OCH 2 CH 2 ) i  group, a (C 1 -C 6 )alkyl-(OCH 2 CH 2 ) i —O(C 1 -C 6 )alkyl group, a (CH 2 CH 2 O) i (C 1 -C 6 )alkyl group, a CH(SO 3 H)—(C 1 -C 6 )alkyl group, a (C 1 -C 6 )alkyl-CH(SO 3 H) group, a (C 1 -C 6 )alkyl-cyclohexyl group, a NR 19 —(C 1 -C 6 )alkyl group, a NR 20 —(CH 2 CH 2 O) i (C 1 -C 6 )alkyl group, a NR 21 -aryl group, a NR 21 -heteroaryl group, a (C 1 -C 6 )alkyl-NR 22 C(═O)—(C 1 -C 6 )alkyl group or a (C 1 -C 6 )alkyl-NR 22 C(═O)—(C 1 -C 6 )alkyl-(OCH 2 CH 2 ) i  group; 
 R 9 , R 20 , R 21  and R 22  each represent, independently of each other, a hydrogen atom or a (C 1 -C 6 )alkyl group; 
 i represents an integer between 1 and 50; and
 AA denotes a natural or unnatural amino acid, of configuration D or L, selected from: alanine (Ala), β-alanine, γ-aminobutyric acid, 2-amino-2-cyclohexylacetic acid, 2-amino-2-phenylacetic acid, arginine (Arg), asparagine (Asn), aspartic acid (Asp), citrulline (Cit), cysteine (Cys), α,α-dimethyl-γ-aminobutyric acid, β,β-dimethyl-γ-aminobutyric acid, glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), ε-acetyl-lysine (AcLys), methionine (Met), ornithine (Orn), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), valine (Val). 
 
 
       
     
     
         22 . The Cryptophycin payload of formula (II) according to  claim 21 , wherein the sequence (AA) w  has the formula: 
       
         
           
           
               
               
           
         
         in which R 23  represents the side chain of AA. 
       
     
     
         23 - 24 . (canceled) 
     
     
         25 . The Cryptophycin payload of formula (II) according to  claim 21 , wherein L2 represents a (C 1 -C 6 )alkyl group or a (C 1 -C 6 )alkyl-(OCH 2 CH 2 ) i  group or a CH(SO 3 H)—(C 1 -C 6 )alkyl group. 
     
     
         26 . (canceled) 
     
     
         27 . A Conjugate of formula (III): 
       
         
           
           
               
               
           
         
         wherein
 R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9  and R 10  are as defined in  claim 1 ; 
 Y is selected from (C 1 -C 6 )alkyl-NR 11 , (C 1 -C 6 )alkyl-O, (C 1 -C 6 )alkyl-S, C(═O)O, C(═O)NH, (C 1 -C 6 )alkyl-C(═O)O, (C 1 -C 6 )alkyl-C(═O)NH or (C 1 -C 6 )alkyl-triazole group; 
 L represents a linker 
 G represents the product of reaction between RCG1, a reactive group present at the end of the linker and RCG2, an orthogonal reactive group present on Ab; and 
 Ab represents an antibody. 
 
       
     
     
         28 . (canceled) 
     
     
         29 . The Conjugate of formula (III) according to  claim 27 , wherein RCG2 is selected from:
 ε-amino groups of lysines borne by the side chains of the lysine residues that are present at the surface of an antibody,   α-amino groups of N-terminal amino acids of antibody heavy and light chains,   saccharide groups of the hinge region,   thiols of cysteines generated by reducing intra-chain disulfide bonds or the thiols of engineered cysteines   amide groups borne by the side chains of some glutamine residues that are present at the surface of an antibody, or   aldehyde groups introduced using formylglycine generating enzyme.   
     
     
         30 - 31 . (canceled) 
     
     
         32 . A Process for preparing the cryptophycin compound of formula (I) comprising the step: 
       
         
           
           
               
               
           
         
         wherein:
 Step (i) is a peptidic coupling between fragments AD3 and alternative BC in the presence of coupling reagents; 
 Step (ii) is a macrocyclization by ring closing metathesis in the presence of a catalyst; 
 Step (iii) is a deprotection of the p-methoxybenzyl ether in acidic conditions; 
 Step (iv) is an oxidation of the alcohol using an oxidizing agent; 
 Step (v) is an introduction of the epoxide by asymmetric Corey-Chaykovsky reaction using appropriately substituted isothiocineole-derived chiral sulfonium in the presence of a base; and 
 Step (vi) is a reduction of the azido group. 
 
       
     
     
         33 . The Process for preparing a conjugate of formula (III) comprising the steps of:
 (i) placing in contact and leaving to react:
 an optionally buffered aqueous solution of an antibody, optionally modified by means of a odifying agent, and 
 a solution of a cryptophycin payload of formula (II) as defined in  claim 17 , wherein 
   the chemical group RCG1 of the cryptophycin payload of formula (II) is reactive towards the chemical group RCG2 present on the antibody especially towards the amino groups present on antibodies, the said chemical groups RCG2 having been introduced, where appropriate, by the modifying agent,   so as to attach the cryptophycin payload of formula (II) to the antibody by formation of a covalent bond; and   (ii) optionally separating the conjugate formed in step (i) from the cryptophycin payload of formula (II) and/or from the unreacted antibody and/or from any aggregates that may have formed.   
     
     
         34 . A medicament, wherein the medicament comprises a conjugate of formula (III). 
     
     
         35 . A pharmaceutical composition, wherein the composition comprises a conjugate of formula (III) and a pharmaceutically acceptable excipient. 
     
     
         36 - 37 . (canceled) 
     
     
         38 . A method of treating cancer in a patient in need thereof, comprising ad mistering a therapeutically effect amount of a compound of formula (I). 
     
     
         39 . A method of treating cancer in a patient in need thereof, comprising administering a therapeutically effect amount of a compound of formula (III).