US20180369401A1 - Novel peptidic linkers and cryptophycin conjugates, their preparation and their therapeutic use - Google Patents

Novel peptidic linkers and cryptophycin conjugates, their preparation and their therapeutic use Download PDF

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US20180369401A1
US20180369401A1 US15/975,423 US201815975423A US2018369401A1 US 20180369401 A1 US20180369401 A1 US 20180369401A1 US 201815975423 A US201815975423 A US 201815975423A US 2018369401 A1 US2018369401 A1 US 2018369401A1
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Hervé Bouchard
Marie-Priscille Brun
Philippe Hubert
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Sanofi SA
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/15—Depsipeptides; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • A—HUMAN NECESSITIES
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    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/65—Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/6811—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin
    • A61K47/6817—Toxins
    • A61K47/6829—Bacterial toxins, e.g. diphteria toxins or Pseudomonas exotoxin A
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6889—Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/06—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K11/00—Depsipeptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
    • C07K5/06052—Val-amino acid

Definitions

  • the present disclosure relates to new peptidic linkers, to new cryptophycin payloads, to new cryptophycin conjugates, to compositions containing them and to their therapeutic use, such as for use as anticancer agents.
  • the present disclosure also relates to the process for preparing these conjugates.
  • linkers are enzymatically cleaved in the lysosome of cells, by enzymes such as Cathepsin B for example.
  • Cathepsin B is a cysteine proteinase belonging to the papain family and one of the main lysosomal proteinases among mammals. It is involved in protein turnover and in the maintenance of cellular metabolism as well as in several other physiological or pathological processes, like for example tumoral progression. Its over-expression, both at the genetic and proteic levels, has been demonstrated in tumors, the increase of protein leading also to an increase of enzymatic activity.
  • a payload is a compound comprising the cytotoxic drug conjugated to a linker—and thus potentially its reactivity towards antibody conjugation and the stability of the subsequent ADC.
  • Increasing payload solubility should allow to increase the DAR, the monomeric purity and ADC stability, especially in terms of aggregation propensity.
  • the disclosure relates to new peptidic compounds, chosen from compounds of formula (I):
  • the disclosure further relates to cryptophycin payloads of formula (IV):
  • the present disclosure further relates to conjugates of formula (V)
  • Each substituent R 1 to R 12 may also adopt one of the spatial configurations (e.g. R or S or alternatively Z or E) as described in the examples.
  • the compounds of formula (I), (II), (Ill), (IV) and (V) may contain at least one asymetric carbon atoms. They may therefore exist in the form of enantiomers or diastereoisomers. These enantiomers and diastereoisomers, and also mixtures thereof, including racemic mixtures, form part of this disclosure.
  • the compounds of formula (I), (II), (IV), including those that are illustrated, may exist in the form of bases or of acid-addition salts, for instance of pharmaceutically acceptable acids.
  • these compounds may exist in the form of SO 3 ⁇ alkali metal salts, such as of SO 3 ⁇ sodium salts (SO 3 ⁇ + Na).
  • Fragments of (conventional) antibodies typically comprise a portion of an intact antibody, such as the antigen binding region or variable region of the intact antibody, and retain the biological function of the conventional antibody.
  • fragments include Fv, Fab, F(ab′)2, Fab′, dsFv, (dsFv)2, scFv, sc(Fv)2, nanobodies and diabodies.
  • a SO 3 H function can be under salt forms such as alkali metal salts, for instance sodium salts (SO 3 ⁇ + Na).
  • ADC antibody-drug conjugate
  • ALK (C 1 -C 12 )alkylene group, for instance (C 1 -C 6 )alkylene, such as of the form —(CH 2 ) n —, n being an integer ranging from 1 to 12 and for example ranging from 1 to 6
  • aq. aqueous
  • Ar argon
  • AUC area under the curve
  • BCN (1 ⁇ ,8 ⁇ ,9 ⁇ )-bicyclo[6.1.0]non-4-yne-9-methanol
  • CHCl 3 chloroform
  • CH 3 CN acetonitrile
  • CO 2 carbon dioxide
  • CR complete response
  • crypto denotes the unit of formula
  • FIG. 1 In vivo efficacy of Ex. 6 against MDA-MB-231 xenograft in SCID mice
  • FIG. 2 In vivo efficacy of Ex. 16, Ex. 19, Ex. 23 and Ex. 32 against MDA-MB-231 xenograft in SCID mice at 2.5 mg/kg
  • FIG. 3 In vivo efficacy of Ex. 16, Ex. 19, Ex. 23 and Ex. 32 against MDA-MB-231 xenograft in SCID mice at 1.25 mg/kg
  • FIG. 4 In vivo efficacy of Ex. 26 against MDA-MB-231 xenograft in SCID mice
  • FIG. 5 In vivo efficacy of Ex. 29 against MDA-MB-231 xenograft in SCID mice
  • FIG. 6 In vivo efficacy of Ex. 35 against MDA-MB-231 xenograft in SCID mice
  • FIG. 7 In vivo efficacy of Ex. 41 against MDA-MB-231 xenograft in SCID mice
  • the disclosure relates to new peptidic compounds, chosen from compounds of formula (I):
  • cation represents for example sodium, potassium or cesium
  • GI represents at least one electroinductive group such as —NO 2 or a halogen atom (-Hal), for instance a fluorine atom (—F).
  • electroinductive group such as —NO 2 or a halogen atom (-Hal), for instance a fluorine atom (—F).
  • —Hal halogen atom
  • F fluorine atom
  • the linker L which is present in formula (I) is of formula (II):
  • the disclosure relates to compounds of formula (I):
  • the disclosure relates to compounds of formula (I):
  • the disclosure relates to compounds of formula (I):
  • the disclosure relates to compounds of formula (I):
  • the disclosure relates to compounds of formula (I):
  • the disclosure relates to compounds of formula (I):
  • (AA)w contains at least one substituted amino acid AA s and L2 represents:
  • (AA)w contains at least one substituted amino acid AA s and L2 represents:
  • (AA)w contains w non-substituted amino acid AA ns and L2 represents:
  • a (C 1 -C 6 )alkyl-NA 7 -(C 1 -C 6 )alkyl group such as a (CH 2 ) 2 —NA 7 -(CH 2 ) 2 group in which A 7 is as defined above.
  • (AA)w contains w non-substituted amino acid AA ns and L2 represents:
  • a (C 1 -C 6 )alkyl-NA 7 -(C 1 -C 6 )alkyl group such as a (CH 2 ) 2 —NA 7 -(CH 2 ) 2 group in which A 7 represents:
  • (AA)w contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents: a NA 7 -(C 1 -C 6 )alkyl group, a (C 1 -C 6 )alkyl-NA 7 group, a NA 7 -(CH 2 CH 2 O) i (C 1 -C 6 )alkyl group, a NA 7 -aryl group, a NA 7 -heteroaryl group, a (C 1 -C 6 )alkyl-NA 7 C( ⁇ O)—(C 1 -C 6 )alkyl group, a (C 1 -C 6 )alkyl-NA 7 C( ⁇ O)—(C 1 -C 6 )alkyl-(OCH 2 CH 2 ) i group, a (C 1 -C 6 )alkyl-C( ⁇ O)NA 7 -(C 1 -C 6 )alkyl-(
  • (AA)w contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • (AA)w contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • (AA)w contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • (AA)w contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 comprises a A 7 representing:
  • each A 7 comprising a SO 3 H function can be under salt forms such as alkali metal salts, for instance sodium salts (SO 3 ⁇ + Na).
  • (AA)w contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • L1 may be one of the following (III1-5):
  • AA denotes an amino acid.
  • An amino acid is a compound of formula NH 2 —CHA 10 -COOH wherein A 10 represents the side chain of the AA.
  • AA can be a substituted AA s or non-substituted AA ns amino acid.
  • a non-substituted amino acid AA ns denotes natural or non-natural amino acid, of configuration D or L, identical to or derived from: alanine (Ala), ⁇ -alanine, ⁇ -aminobutyric acid, 2-amino-2-cyclohexylacetic acid, 2-amino-2-phenylacetic acid, arginine (Arg), asparagine (Asn), aspartic acid (Asp), citrulline (Cit), cysteine (Cys), ⁇ , ⁇ -dimethyl- ⁇ -aminobutyric acid, ⁇ , ⁇ -dimethyl- ⁇ -aminobutyric acid, glutamine (Gln), glutamic acid (Glu), glycine (Gly), homo-cysteine, selenocysteine, homo-selenocysteine, histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), ⁇ -acety
  • AA ns represents alanine (Ala), citrulline (Cit), glycine (Gly), isoleucine (lie), leucine (Leu), lysine (Lys), ⁇ -acetyl-lysine (AcLys), phenylalanine (Phe), tryptophan (Trp) and valine (Val).
  • the substituted amino acids AA s have the formula (VI):
  • the substituted amino acids AA s have the formula (VI):
  • the substituted amino acids AA s have the formula (VI):
  • the substituted amino acids AA s have the formula (VI):
  • T examples of T that may be mentioned include:
  • the substituted amino acids AA s have the formula (VI) wherein:
  • substituted amino acids AAs have the formula (VI) wherein:
  • substituted amino acids AA s have the formula (VI) wherein:
  • the sequence (AA) w has the formula:
  • a 10 represents the side chain of one of the substituted AA s or non-substituted AA s amino acids described above.
  • sequences of non-substituted amino acids (AA s )w are as follows: Gly-Gly, Phe-Lys, Val-Lys, Val-AcLys, Val-Cit, Phe-Phe-Lys, D-Phe-Phe-Lys, Gly-Phe-Lys, Ala-Lys, Val-Ala, Phe-Cit, Phe-Gly, Leu-Cit, Ile-Cit, Trp-Cit, Phe-Ala, Ala-Phe, Gly-Gly-Gly, Gly-Ala-Phe, Gly-Phe-Gly, Gly-Val-Cit, Gly-Phe-Leu-Cit (SEQ ID No. 3), Gly-Phe-Leu-Gly (SEQ ID No. 4), Ala-Leu-Ala-Leu (SEQ ID No. 5).
  • the sequence (AA ns )w of non-substituted amino acids AA ns is selected from the following list: Gly-Gly, Phe-Lys, Val-Lys, Val-AcLys, Val-Cit, Phe-Phe-Lys, D-Phe-Phe-Lys, Gly-Phe-Lys, Ala-Lys, Val-Ala, Phe-Cit, Phe-Gly, Leu-Cit, Ile-Cit, Trp-Cit, Phe-Ala, Ala-Phe, Gly-Gly-Gly, Gly-Ala-Phe, Gly-Phe-Gly, Gly-Val-Cit, Gly-Phe-Leu-Cit (SEQ ID No. 3), Gly-Phe-Leu-Gly (SEQ ID No. 4), and Ala-Leu-Ala-Leu (SEQ ID No. 5), such as Val-A
  • sequence (AA s )w containing at least one substituted amino acid AA s is selected from the list:
  • linkers L of formula (II) that are the subject matter of the disclosure, mention may be made for example of the following compounds:
  • RCG1 examples include:
  • RCG1 may be:
  • R a Z a — may represent HO—, CH 3 O—, CH 2 ⁇ CH—CH 2 O—,
  • cation represents for example sodium, potassium or cesium or
  • GI represents at least one electroinductive group such as —NO 2 or a halogen atom, such as a fluorine atom (F).
  • electroinductive group such as —NO 2 or a halogen atom, such as a fluorine atom (F).
  • F fluorine atom
  • RaZ a —C( ⁇ O)— group is the following:
  • RCG1 may be chosen from one of those described in the examples that is to say chosen from the following groups:
  • the present disclosure further relates to cryptophycin payloads of formula (IV):
  • Each substituent R 1 to R 12 may also adopt one of the spatial configurations (e.g. R or S or alternatively Z or E) as described in the examples.
  • the compounds of formula (IV) may contain at least one asymmetric carbon atom. They may therefore exist in the form of enantiomers or diastereoisomers. These enantiomers and diastereoisomers, and also mixtures thereof, including racemic mixtures, form part of the disclosure.
  • the cryptophycin compound of formula (IV) may be one of the cryptophycin compounds of formula (I) described in WO2011/001052 (such as one of D 1 -D 8 ) or in PCT/EP2016/076603, (such as one of D 1 -D 19 ), as mentioned above.
  • cryptophycin compound may be an equivalent unit described in one of the examples.
  • a group of compounds is composed of the compounds for which R 1 represents a (C 1 -C 6 )alkyl, such as a methyl group.
  • a group of compounds is composed of the compounds for which each of R 2 and R 3 represents a hydrogen atom.
  • a group of compounds is composed of the compounds for which one of R 2 and R 3 represents a (C 1 -C 6 )alkyl, such as a methyl group, and the other one represents a hydrogen atom.
  • a group of compounds is composed of the compounds for which R 2 and R 3 form together with the carbon atom to which they are attached a (C 3 -C 6 )cycloalkyl group, such as a cyclopropyl group.
  • a group of compounds is composed of the compounds for which each of R 4 and R 5 represents a (C 1 -C 6 )alkyl, such as a methyl group.
  • a group of compounds is composed of the compounds for which X represents an oxygen atom, that is to say O.
  • a group of compounds is composed of the compounds for which X represents NH.
  • a group of compounds is composed of the compounds for which R 7 and R 8 represent independently of each other an hydrogen atom or a (C 1 -C 6 )alkyl group, such as an isobutyl group or a neopentyl group.
  • a group of compounds is composed of the compounds for which R 9 represents two substituents selected from a methoxy group and a chlorine atom, such as the two R 9 substituents are 3-Cl and 4-methoxy.
  • the phenyl nucleus comprises two substituents in positions 3 and 4 on the phenyl nucleus.
  • a group of compounds is composed of the compounds for which R 10 represents a hydrogen atom.
  • a group of compounds is composed of the compounds for which Y is positioned in the para position of the phenyl nucleus.
  • a group of compounds is composed of the compounds for which Y represents NR 11 —(C 1 -C 6 )alkyl, such as NR 11 —(C 1 -C 3 )alkyl, for instance NH—CH 2 .
  • a group of compounds is composed of the compounds for which L of formula (II) comprises at least one substituted amino acid AA s in the sequence of w amino acids (AA)w, L1 and L2 are as defined in formula (II).
  • a group of compounds is composed of the compounds for which L of formula (II) comprises a sequence of w non-substituted amino-acid AA ns , L1 and L2 are as defined in formula (II).
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and L2 represents:
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and L2 represents:
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains w non-substituted amino acid AA ns and L2 represents:
  • a (C 1 -C 6 )alkyl-NA 7 -(C 1 -C 6 )alkyl group such as a (CH 2 ) 2 —NA 7 -(CH 2 ) 2 group in which A 7 is as defined above.
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains w non-substituted amino acid AA ns and L2 represents:
  • a (C 1 -C 6 )alkyl-NA 7 -(C 1 -C 6 )alkyl group such as a (CH 2 ) 2 —NA 7 -(CH 2 ) 2 group in which A 7 represents:
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a preferred group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a more preferred group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a still more preferred group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a most preferred group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 comprises a A 7 representing:
  • each A 7 comprising a SO 3 H function can be under salt forms such as alkali metal salts, for instance sodium salts (SO 3 ⁇ + Na).
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a group of compounds is composed of the compounds comprising in L a sequence (AA s )w containing at least one substituted amino acid AA s , (AA s )w being selected from the list:
  • a group of compounds is composed of the compounds of the following structure (beta epoxide configuration):
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , n, RCG 1 , X, Y and L are as defined in formula (IV).
  • the present disclosure relates to compounds of formula (IV) wherein:
  • the attachment between the cryptophycin payload of formula (IV) and the antibody, in order to obtain the conjugate of formula (V), is produced by the reaction between the reactive chemical group RCG1 present on the payload as defined above that is reactive towards a reactive group RCG2 present on the polypeptide such as the antibody.
  • the reaction between RCG1 and RCG2 ensures the attachment of the compound of formula (IV) to the antibody by formation of a covalent bond.
  • parts of RCG1 and RCG2 can remain forming the attachment between the linker and the antibody.
  • RCG2 examples include (Garnett M. C., et al., Advanced Drug Delivery Reviews 2001, 53, 171-216):
  • RCG1 is of the type (ii) above
  • RCG2 is of the type (ii) above
  • RCG2 it is possible to chemically modify the antibody using an adequate modifying agent or to introduce one or more unnatural amino acids so as to introduce the adequate functions RCG2.
  • Examples of G that may be mentioned include
  • G represents the following groups:
  • a group of compounds is composed of the compounds for which R 1 represents a (C 1 -C 6 )alkyl, such as a methyl group.
  • a group of compounds is composed of the compounds for which each of R 2 and R 3 represents a hydrogen atom.
  • a group of compounds is composed of the compounds for which one of R 2 and R 3 represents a (C 1 -C 6 )alkyl group, such as a methyl group and the other one represents a hydrogen atom.
  • a group of compounds is composed of the compounds for which R 2 and R 3 form together with the carbon atom to which they are attached a (C 3 -C 6 )cycloalkyl group, such as a cyclopropyl group.
  • a group of compounds is composed of the compounds for which each of R 4 and R 5 represents a (C 1 -C 6 )alkyl group, in particular a methyl group.
  • a group of compounds is composed of the compounds for which X represents an oxygen atom, that is to say O.
  • a group of compounds is composed of the compounds for which X represents NH.
  • a group of compounds is composed of the compounds for which R 7 and R 8 represent independently of each other a hydrogen atom or a (C 1 -C 6 )alkyl group, such as an isobutyl group or a neopentyl group.
  • a group of compounds is composed of the compounds for which R 9 represents two substituents selected from a methoxy group and a chlorine atom.
  • the phenyl nucleus comprises two substituents in positions 3 and 4 on the phenyl nucleus. For example 3-Cl and 4-methoxy.
  • a group of compounds is composed of the compounds for which R 10 represents a hydrogen atom.
  • a group of compounds is composed of the compounds for which Y is positioned in the para position of the phenyl nucleus.
  • a group of compounds is composed of the compounds for which Y NR 11 —(C 1 -C 6 )alkyl, such as NR 11 —(C 1 -C 3 )alkyl, for instance NH—CH 2 .
  • a group of compounds is composed of the compounds for which L of formula (II) comprises at least one substituted amino acid AA s in the sequence of w amino acids (AA)w, L1 and L2 are as defined in formula (II).
  • a group of compounds is composed of the compounds for which L of formula (II) comprises a sequence of w non-substituted amino-acid AA ns , L1 and L2 are as defined in formula (II).
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and L2 represents:
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and L2 represents:
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains w non-substituted amino acid AA ns and L2 represents:
  • a (C 1 -C 6 )alkyl-NA 7 -(C 1 -C 6 )alkyl group such as a (CH 2 ) 2 —NA 7 -(CH 2 ) 2 group in which A 7 is as defined above.
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains w non-substituted amino acid AA ns and L2 represents:
  • a (C 1 -C 6 )alkyl-NA 7 -(C 1 -C 6 )alkyl group such as a (CH 2 ) 2 —NA 7 -(CH 2 ) 2 group in which A 7 represents:
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a preferred group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a more preferred group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a still more preferred group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a most preferred group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 comprises a A 7 representing:
  • each A 7 comprising a SO 3 H function can be under salt forms such as alkali metal salts, for instance sodium salts (SO 3 +Na).
  • a group of compounds is composed of the compounds for which (AA)w in L of formula (II) contains at least one substituted amino acid AA s and/or w non-substituted amino acid AA ns and L2 represents:
  • a group of compounds is composed of the compounds comprising in L a sequence (AA s )w containing at least one substituted amino acid AA s , (AA s )w being selected from the list:
  • a group of compounds is composed of the compounds for which Ab is an anti-Epha2 antibody, for instance hu2H11_R35R74 antibody as represented by SEQ ID NO: 1 (light chain of antibody hu2H11_R35R74) and by SEQ ID NO: 2 (heavy chain of antibody hu2H11_R35R74) corresponding to respectively SEQ ID NO: 16 and SEQ ID NO:18 represented in WO2011039724 A1.
  • the present disclosure relates to compounds of formula (V) wherein:
  • a group of compounds is composed of the compounds of the following structure (beta epoxide configuration):
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , n, RCG 1 , X, Y and L are as defined in formula (IV).
  • the compounds of general formula (I), (II), (Ill), (IV) and (V) can be prepared by the following processes.
  • Scheme 1 depicted the synthesis starting with Boc- ⁇ -Ala-OH (CAS number [3303-84-2]) but may also apply to other Boc protected amino-alkyl acids which are commercially available for n ranging from 3 to 10. It depicted the synthesis using succinic anhydride but may also apply to glutaric anhydre or alkyl diacids which are commercially available for n ranging from 3 to 10.
  • Scheme 2 depicted the synthesis of linkers using a Val-Ala dipeptide but may also apply to other dipeptides; it depicted the synthesis using spacers 3 or 4 but may also apply to spacers 1 or 2; it depicted the synthesis using Boc-monoprotected ethylenediamine but may also apply to other Boc-monoprotected diamines which are commercially available for n ranging from 3 to 10.
  • Scheme 3 depicted the synthesis of dipeptides starting with Val but may apply to other amino acids listed above; it depicted the synthesis using L-Glu tert-butyl ester (CAS number [45120-30-7]) but may also apply to other amino acids bearing a carboxylic acid on their side chain such as, for example, L-Asp tert-butyl ester (CAS number [4125-93-3]), D-Asp tert-butyl ester (CAS number [148823-36-3]), D-Glu tert-butyl ester (CAS number [25456-76-2]), 2-amino-hexanedioic acid 1-tert-butyl ester (CAS number [1245806-58-9]) or 2-amino-heptanedioic acid 1-tert-butyl ester (CAS number [1888498-03-0]).
  • L-Asp tert-butyl ester CAS number [4125-93-3]
  • Scheme 4 depicted the synthesis of dipeptides using p-aminobenzyl alcohol (CAS number [623-04-1]) but may also apply to other aminobenzyl alcohol compounds which are commercially available such as, for example, 4-(1-hydroxyethyl)-aniline (racemic (CAS number [14572-89-5]) or enantiopure (R) (CAS number [210754-25-9]) or (S) (CAS number [500229-84-5])), 4-amino- ⁇ , ⁇ -dimethyl-benzene-methanol (CAS number [23243-04-1]), 4-amino- ⁇ -methoxy- ⁇ -methyl-benzenemethanol (CAS number [1379318-81-6]), 4-amino- ⁇ -methyl- ⁇ -trifluoromethyl-benzenemethanol (CAS number [851652-56-7]), 2-amino-benzenemethanol (CAS number [5344-90-1]), 2-amino- ⁇ -methyl-benzenemethanol (racemic (CAS number [10517
  • dipeptide 5 can be prepared following the synthesis described in Scheme 6.
  • Scheme 7 depicted the synthesis of dipeptides using p-aminobenzyl alcohol (CAS number [623-04-1]) but may also apply to other aminobenzyl alcohol compounds which are commercially available such as, for example, 4-(1-hydroxyethyl)-aniline (racemic (CAS number [14572-89-5]) or enantiopure (R) (CAS number [210754-25-9]) or (S) (CAS number [500229-84-5])), 4-amino- ⁇ , ⁇ -dimethyl-benzene-methanol (CAS number [23243-04-1]), 4-amino- ⁇ -methoxy- ⁇ -methyl-benzenemethanol (CAS number [1379318-81-6]), 4-amino- ⁇ -methyl- ⁇ -trifluoromethyl-benzenemethanol (CAS number [851652-56-7]), 2-amino-benzenemethanol (CAS number [5344-90-1]), 2-amino- ⁇ -methyl-benzenemethanol (racemic (CAS number [10517
  • Cryptophycin compounds may be prepared as described in WO2011/001052 for X ⁇ O and in PCT/EP2016/076603 for X ⁇ NH.
  • coupling reagents such as, for example, EDC and HOBt
  • new cryptophycin payloads bearing a spacer with improved hydrophilicity may also be prepared as depicted in Scheme 9.
  • Schemes 8 and 9 depicted the synthesis of payloads using the p-benzylic amine of C52 but may also apply to other cryptophycin compounds; it depicted the synthesis using Val-Ala dipeptides but may also apply to other dipeptides; it depicted the synthesis using spacers 1 or 2 but may also apply to spacers 3 or 4.
  • Scheme 10 depicted the synthesis of payloads using Val-Ala dipeptides but may also apply to other dipeptides; it depicted the synthesis using p-aminobenzyl alcohol (CAS number [623-04-1]) but may also apply to other aminobenzyl alcohol compounds which are commercially available such as, for example, 4-(1-hydroxyethyl)-aniline (racemic (CAS number [14572-89-5]) or enantiopure (R) (CAS number [210754-25-9]) or (S) (CAS number [500229-84-5])), 4-amino- ⁇ , ⁇ -dimethyl-benzene-methanol (CAS number [23243-04-1]), 4-amino- ⁇ -methoxy- ⁇ -methyl-benzenemethanol (CAS number [1379318-81-6]), 4-amino- ⁇ -methyl- ⁇ -trifluoromethyl-benzenemethanol (CAS number [851652-56-7]), 2-amino-benzenemethanol (CAS number [5344-
  • coupling reagents such as, for example, EDC and HOBt and deprotection of the Fmoc amine in the presence of a base such as, for example, piperidine
  • Scheme 11 depicted the synthesis of payloads using the p-benzylic amine of C52 but may also apply to other cryptophycin compounds; it depicted the synthesis using dipeptide 1 but may also apply to dipeptide 5; it depicted the synthesis using glutaric anhydride but may also apply to succinic anhydride or alkyl diacids which are commercially available for n ranging from 3 to 10.
  • a base such as, for example, DIEA
  • Scheme 12 depicted the synthesis of payloads using the p-benzylic amine of C52 but may also apply to other cryptophycin compounds; it depicted the synthesis using dipeptide 6 but may also apply to dipeptide 2.
  • Scheme 13 depicted the synthesis of payloads using the p-benzylic amine of C52 but may also apply to other cryptophycin compounds; it depicted the synthesis using dipeptide 5 but may also apply to dipeptide 1; it depicted the synthesis using spacers 3 or 4 but may also apply to spacers 1 or 2.
  • Scheme 14 depicted the synthesis of payloads using the p-benzylic amine of C52 but may also apply to other cryptophycin compounds; it depicted the synthesis using dipeptide 6 but may also apply to dipeptide 2; it depicted the synthesis using spacers 3 or 4 but may also apply to spacers 1 or 2.
  • a base such as, for example, DIEA
  • a base such as, for example, D
  • Scheme 15 depicted the synthesis of payloads using the activated p-benzylic alcohol of C52 but may also apply to other cryptophycin compounds; it depicted the synthesis using Val-Ala dipeptides but may also apply to other dipeptides; it depicted the synthesis using Boc-monoprotected ethylenediamine but may also apply to other Boc-monoprotected diamines which are commercially available for n ranging from 3 to 10; it depicted the synthesis using spacers 1 or 2 but may also apply to spacers 3 or 4.
  • new cryptophycin payloads of formula (IV) bearing a spacer with improved hydrophilicity may also be prepared as depicted in Scheme 16.
  • a base such as, for example, DIEA
  • a catalyst such as, for example, tetrakis-(triphenylphosphine)palladium
  • Scheme 16 depicted the synthesis of payloads using the activated p-benzylic alcohol of C52 but may also apply to other cryptophycin compounds.
  • Scheme 17 depicted the synthesis of payloads using Val-Ala dipeptides but may also apply to other dipeptides; it depicted the synthesis using p-aminobenzyl alcohol (CAS number [623-04-1]) but may also apply to other aminobenzyl alcohol compounds which are commercially available such as, for example, 4-(1-hydroxyethyl)-aniline (racemic (CAS number [14572-89-5]) or enantiopure (R) (CAS number [210754-25-9]) or (S) (CAS number [500229-84-5])), 4-amino- ⁇ , ⁇ -dimethyl-benzene-methanol (CAS number [23243-04-1]), 4-amino- ⁇ -methoxy- ⁇ -methyl-benzenemethanol (CAS number [1379318-81-6]), 4-amino- ⁇ -methyl- ⁇ -trifluoromethyl-benzenemethanol (CAS number [851652-56-7]), 2-amino-benzenemethanol (CAS number [5344-
  • Scheme 18 depicted the synthesis of payloads using the activated p-benzylic alcohol of C52 but may also apply to other cryptophycin compounds; it depicted the synthesis using dipeptide 7 but may also apply to dipeptide 3; it depicted the synthesis using glutaric anhydride but may also apply to succinic anhydride or alkyl diacids which are commercially available for n ranging from 3 to 10.
  • Scheme 19 depicted the synthesis of payloads using the activated p-benzylic alcohol of C52 but may also apply to other cryptophycin compounds; it depicted the synthesis using dipeptide 8 but may also apply to dipeptide 4; it depicted the synthesis using glutaric anhydride but may also apply to succinic anhydride or alkyl diacids which are commercially available for n ranging from 3 to 10.
  • Scheme 20 depicted the synthesis of payloads using the p-benzylic amine of C52 but may also apply to other cryptophycin compounds; it depicted the synthesis using dipeptide 7 but may also apply to dipeptide 3; it depicted the synthesis using spacers 3 or 4 but may also apply to spacers 1 or 2.
  • Scheme 21 depicted the synthesis of payloads using the p-benzylic amine of C52 but may also apply to other cryptophycin compounds; it depicted the synthesis using dipeptide 8 but may also apply to dipeptide 4; it depicted the synthesis using spacers 3 or 4 but may also apply to spacers 1 or 2.
  • step (ii) the conjugate of formula (V) from step (i) is separated only from the unreacted cryptophycin payload of formula (IV) and from any aggregates formed, and any unreacted antibody is left in the solution.
  • the function of the placing in contact is to react the chemical groups RCG1 and RCG2 in order to ensure attachment of the cryptophycin payload of formula (IV) to the polypeptide such as the antibody by formation of a covalent bond; such as,
  • aggregates means associations that may form between two or more antibodies, the antibodies possibly having been modified by conjugation. Aggregates are liable to form under the influence of a wide variety of parameters such as a high concentration of antibody in the solution, the pH of the solution, high shear forces, the number of grafted drugs and their hydrophobic nature, the temperature (see the references cited in the introduction of J. Membrane Sci. 2008, 318, 311-316), the influence of some of them, however, having not been clearly elucidated. In the case of proteins or antibodies, reference may be made to AAPS Journal , “Protein Aggregation and Bioprocessing” 2006, 8(3), E572-E579. The aggregate content may be determined via known techniques such as SEC (see in this respect Analytical Biochemistry 1993, 212 (2), 469-480).
  • the aqueous solution of the antibody may be buffered with buffers for example, potassium phosphate or HEPES or a mixture of buffers such as buffer A described later.
  • the buffer depends on the nature of the antibody.
  • the cryptophycin payload of formula (IV) is dissolved in a polar organic solvent such as DMSO or DMA.
  • the reaction takes place at a temperature generally ranging from 20° C. to 40° C.
  • the reaction time may be ranging from 1 to 24 hours.
  • the reaction between the antibody and the cryptophycin payload of formula (IV) may be monitored by SEC with a refractometric and/or ultraviolet detector and/or HRMS in order to determine its degree of progress. If the degree of substitution is insufficient, the reaction can be left for longer and/or cryptophycin compound can be added.
  • the conjugate may be purified, for example, by steric exclusion chromatography (SEC), by adsorption chromatography (for instance ion exchange, IEC), by hydrophobic interaction chromatography (HIC), by affinity chromatography, by chromatography on mixed supports such as ceramic hydroxyapatite, or by HPLC. Purification by dialysis or diafiltration may also be used.
  • SEC steric exclusion chromatography
  • IEC adsorption chromatography
  • HIC hydrophobic interaction chromatography
  • HPLC hydrophobic interaction chromatography
  • purification by dialysis or diafiltration may also be used.
  • the solution of the conjugate may undergo an ultrafiltration and/or diafiltration step (iii). After these steps, the conjugate in aqueous solution is thus obtained.
  • the antibody can be a monoclonal antibody selected from the group consisting of a murine, chimeric, a humanized and a human antibody.
  • the antibody is a monospecific antibody, i.e. an antibody specifically binding to one single target.
  • it might be a multispecific antibody.
  • the antibody is a IgG antibody, for instance an IgG 1 , an IgG 2 , an IgG 3 or an IgG 4 antibody.
  • the antibody according to the invention specifically binds to a target, thereby directing the biologically active compound as a cytotoxic compound towards said target.
  • “specifically binds” or “binds specifically to” or “binds to” or the like means that an antibody or antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiological conditions.
  • Specific binding can be characterized by an equilibrium dissociation constant (K D ) of at least about 1 ⁇ 10 ⁇ 8 M or less (e.g., a smaller K D denotes a tighter binding).
  • K D equilibrium dissociation constant
  • Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like.
  • antibodies have been characterized, for example, by their specific binding to target and/or target antigen using surface plasmon resonance, e.g., BIACORETM.
  • the target typically corresponds to a protein expressed at the cell surface, e.g. a protein expressed at the surface of tumour cells.
  • the target is the EphA2 receptor.
  • the EphA2 receptor is an Ephrin receptor, and is also referred to as “Eph receptor A2” or “Epithelial Cell Receptor Protein-Tyrosine kinase”.
  • the antibody specifically binding to the EphA2 receptor might for instance correspond to one of the antibodies described in WO2008/010101 or WO2011/039724.
  • the antibody may optionally be modified with a modifying agent so as to promote the attachment of the cryptophycin payload as previously described.
  • the antibody may especially be monoclonal, polyclonal or multispecific. It may also be an antibody fragment. It may also be a murine, human, humanized or chimeric antibody.
  • the antibody used in the examples of the present invention is hu2H11_R3574, an antagonist antibody against EphA2 receptor.
  • hu2H11_R3574 The sequence of hu2H11_R3574 is represented by SEQ ID NO: 1 (light chain of antibody hu2H11_R35R74) and by SEQ ID NO:2 (heavy chain of antibody hu2H11_R35R74) which correspond to respectively SEQ ID NO: 16 and SEQ ID NO:18 represented in WO2011039724 A1.
  • a conjugate generally comprises from about 1 to 10 cryptophycin compounds covalently attached to the antibody (this is the degree of grafting or “drug-to-antibody ratio” or “DAR”). This number varies as a function of the nature of the antibody and of the cryptophycin compound, and also of the operating conditions used in the conjugation process (for example the number of equivalents of cryptophycin compound relative to the antibody, the reaction time, the nature of the solvent and of any cosolvent). Placing of the antibody and the cryptophycin compound in contact leads to a mixture comprising several conjugates that are individually distinguished from each other by different DARs; optionally the unreacted antibody; optionally aggregates. The DAR that is determined on the final solution thus corresponds to an average DAR.
  • the DAR may be calculated from the deconvolution of the SEC-HRMS spectrum of the conjugate.
  • the DAR (HRMS) is for example greater than 0.5, for instance ranging from 1 to 10, such as ranging from 2 to 7.
  • the conjugate may be used as an anticancer agent. Owing to the presence of the antibody, the conjugate is made highly selective towards tumor cells rather than healthy cells. This makes it possible to direct the cryptophycin compound in an environment similar thereto or directly therein. It is possible to treat solid or liquid cancers.
  • the conjugate may be used alone or in combination with at least one other anticancer agent.
  • the conjugate is formulated in the form of a buffered aqueous solution at a concentration generally ranging from 1 to 10 mg/mL.
  • This solution may be injected in perfusion form per se or may be rediluted to form a perfusion solution.
  • Chromatographic conditions were the following: Column: ACQUITY BEH C18-1.7 ⁇ m-2.1 ⁇ 50 mm; solvents: A: H 2 O (0.1% formic acid), B: CH 3 CN (0.1% formic acid); column temperature: 50° C.; flow rate: 0.8 mL/min; gradient (2.5 min): from 5 to 100% of B in 1.8 min; 2.4 min: 100% of B; 2.45 min: from 100 to 5% of B in 0.05 min.
  • Chromatographic conditions were the following: Column: ACQUITY BEH C18-1.7 ⁇ m-2.1 ⁇ 50 mm; solvents: A: H 2 O (0.1% formic acid), B: CH 3 CN (0.1% formic acid); column temperature: 50° C.; flow rate: 0.6 mL/min; gradient (2 min): from 5 to 50% of B in 1 min; from 50 to 100% of B in 0.3 min; 100% of B during 0.15 min; from 100 to 5% de B in 0.3 min and 5% of B during 0.25 min.
  • Chromatographic conditions were the following: Column: ACQUITY BEH C18-1.7 ⁇ m-2.1 ⁇ 50 mm; solvents: A: H 2 O (0.1% formic acid), B: CH 3 CN (0.1% formic acid); column temperature: 50° C.; flow rate: 0.8 mL/min; gradient (5 min): from 5 to 100% of B in 4.2 min; 4.6 min: 100% of B; 4.8 min: 5% of B.
  • Chromatographic conditions were the following: Column: ACQUITY BEH C18-1.7 ⁇ m-2.1 ⁇ 100 mm; solvents: A: H 2 O (0.1% formic acid), B: CH 3 CN (0.1% formic acid); column temperature: 45° C.; flow rate: 0.6 mL/min; gradient (5.3 min): 5% of B from 0 to 0.3 min, 4 min: 100% of B; 4.6 min: 100% of B; 5.3 min: 5% of B.
  • Chromatographic conditions were the following: Column: ACQUITY BEH C18-1.7 ⁇ m-2.1 ⁇ 50 mm; solvents: A: H 2 O (0.1% formic acid), B: CH 3 CN (0.1% formic acid); column temperature: 45° C.; flow rate: 0.8 mL/min; gradient (10 min): from 5 to 100% of B in 8.6 min; 9.6 min: 100% of B; 9.8 min: 5% of B.
  • the 1 H NMR spectra were acquired on a Bruker Avance spectrometer, either of model DRX-300, DRX-400 or DRX-500.
  • the chemical shifts ( ⁇ ) are given in ppm.
  • the chromatographic analysis was performed on an Agilent HP1100 machine and a Waters BEH SEC 200 1.7 ⁇ m (2.1 ⁇ 150 mm) column at 30° C. with a flow rate of 0.5 mL/min and an isocratic elution of (A) 25 mM ammonium formate+1% formic acid/(B) CH3CN+0.1% formic acid 70/30 for 15 minutes.
  • the mass spectrometry was performed on a Waters QTOF-II machine with electrospray ionization in positive mode (ES+). The mass spectra were deconvoluted with the Waters MaxEnt1 software.
  • the analysis was performed on a Waters Alliance HPLC system or a Hitachi Lachrom HPLC system equipped with a photodiode array detector and a Tosoh Bioscience TSKgel G3000 SWXL 5 ⁇ m column (7.8 ⁇ 300 mm) with a flow rate of 0.5 mL/min and an isocratic elution of 30 minutes with a pH 7 buffer containing 0.2 M of KCl, 0.052 M of KH 2 PO 4 , 0.107 M of K 2 HPO 4 and 20% by volume of isopropanol.
  • a solution of antibody in an aqueous buffer composed of a 96:4 mixture of buffer A and 1 N HEPES was treated with an excess (5 to 10 equivalents) of a solution at approximatively 10 mM of cryptophycin payload in DMA such that the final antibody concentration is 3 mg/mL and the percentage of DMA in the aqueous buffer is 20%.
  • the mixture was analysed by SEC-HRMS to determine the DAR on the population of monomeric antibodies. If the DAR was found insufficient ( ⁇ 3.5-4), the mixture was treated with a further excess (1 to 5 equivalents) of cryptophycin solution in DMA for 2 to 4 additional hours at RT under stirring.
  • the mixture was purified by gel filtration using a Superdex 200 pg matrix (HiLoad 16/60 or 26/60 desalting column, GEHealthcare) pre-equilibrated in aqueous buffer pH 6.5 (buffer B or DPBS) containing 10 to 20% of NMP or a SephadexTM G25 matrix (Hiprep 26/10 desalting column, GEHealthcare) pre-equilibrated in aqueous buffer pH 6.5 (buffer B or DPBS) containing 5 to 10% of NMP.
  • the fractions containing the monomeric conjugated antibody were pooled and concentrated on Amicon Ultra-15 (10 k or 50 k Ultracel membrane, Millipore) to a concentration of between 2 and 5 mg/mL.
  • a buffer exchange or a dilution in the appropriate buffer was then performed to formulate the conjugate in the final buffer.
  • a buffer exchange it was realized by gel filtration using a SephadexTM G25 matrix (NAP-5, NAP-10, NAP-25/PD-10 or Hiprep 26/10 desalting columns, GEHealthcare) pre-equilibrated with the final aqueous buffer whose composition and pH are suited to each conjugate.
  • the conjugate was finally filtered through a Steriflip® filter unit (0.22 ⁇ m Durapore® PVDF membrane, Millipore).
  • the final conjugate was assayed by UV spectrometry or SEC-HPLC so as to measure the conjugate concentration, by SEC-HPLC so as to determine the monomeric purity and by SEC-HRMS so as to determine the DAR from the deconvolution of the mass spectrum of the conjugate.
  • Compound 8 15-(3-(allyloxy)-3-oxopropyl)-14-oxo-2,5,8,11-tetraoxa-15-azaoctadecan-18-oic acid
  • reaction medium was stirred for 3 h at RT. Then H 2 O (20 mL) was added and the mixture was extracted with DCM (2 ⁇ 15 mL). The combined organic phases were washed with brine, dried over MgSO 4 , filtered, concentrated in vacuo and purified by flash chromatography on 5 g of silica gel (gradient elution DCM/MeOH) to give 200 mg of compound 10 (85%).
  • Example 1 15-(3-(((S)-1-(((S)-1-((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)-14-oxo-2,5,8,11-tetraoxa-15-azaoctadecan-18-oic acid
  • example 1 (30 mg, 24.39 ⁇ mol) in THF (5 mL) were added DSC (7.65 mg, 29.27 ⁇ mol) and DIEA (9.88 ⁇ L, 58.54 ⁇ mol). The reaction medium was stirred at RT overnight then concentrated in vacuo and purified by flash chromatography on 5 g of diol modified silica gel (gradient elution DCM/iPrOH) to give 17 mg of example 2 (53%).
  • example 3 15 mg of hu2H11_R35-74 were reacted with 55.2 ⁇ L of a 9.1 mM solution of example 2 in DMA (10 eq.) for 3 h 30.
  • DMA 10 eq.
  • 3.36 mg of example 3 were obtained as a colorless limpid solution at a concentration of 2.24 mg/mL with a DAR of 3.4 (HRMS), a monomeric purity of 99.8% and a global yield of 22%.
  • reaction medium was concentrated in vacuo and purified by flash chromatography on 5 g of C18-grafted silica gel (gradient elution CH 3 CN/H 2 O) to give 12.5 mg of example 5 (73%).
  • Compound 18 (5S,8S)-13-(3-(allyloxy)-3-oxopropyl)-8-isopropyl-2,2,5-trimethyl-4,7,10,14,17-pentaoxo-3-oxa-6,9,13,18-tetraazaicosane-20-sulfonic acid
  • reaction medium was stirred for 1 h at RT, concentrated in vacuo and purified by flash chromatography on 10 g of C18-modified silica gel (gradient elution MeCN/H 2 O) to give 30 mg of compound 20 as a white lacquer (24%).
  • reaction medium was stirred for 1 h at RT and purified by reverse phase chromatography on 1 g of C18-modified silica gel (gradient elution MeCN/H 2 O) to give 15 mg of a white powder that was further purified on silica gel (gradient elution DCM/MeOH/H 2 O) to give 7 mg of example 7 as a white lacquer (24%).
  • reaction medium was stirred at RT under Ar overnight. Then piperidine (142.9 ⁇ L, 1.43 mmol) was added and the reaction medium stirred for 1 h at RT. After concentrating in vacuo, the crude medium was purified by flash chromatography on 5 g of silica gel (gradient elution DCM/MeOH) to give 100 mg of compound 23 as a white solid (73%).
  • Example 8 15-(3-(((S)-1-(((S)-1-((4-((((4-(((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)oxy)methyl)-phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)-14-oxo-2,5,8,11-tetraoxa-15-azaoctadecan-18-oic acid
  • example 9 To a solution of example 8 (49 mg, 35.5 ⁇ mol) in DMF (2 mL), were added, under Ar, DSC (13.9 mg, 53.3 ⁇ mol) and DIEA (9.0 ⁇ L, 53.3 ⁇ mol). The reaction medium was stirred at RT overnight. Additional DSC (13.9 mg, 53.3 ⁇ mol) and DIEA (9.0 ⁇ L, 53.3 ⁇ mol) were added and the stirring carried over 8 h. The reaction medium was concentrated in vacuo and purified by flash chromatography on 10 g of diol-modified silica gel (gradient elution DCM/iPrOH) to give 20 mg of example 9 as a white solid (38%).
  • RMN 1 H (400 MHz, ⁇ in ppm, DMSO-d6): 50:50 conformer mixture; 1.39 (s, 4.5H); 1.40 (s, 4.5H); 2.22 to 2.75 (partially masked m, 6H); 3.23 (s, 3H); 3.38 to 3.63 (m, 30H); 4.55 (m, 2H); 5.19 to 5.36 (m, 2H); 5.90 (m, 1H).
  • Compound 29 24-(3-(allyloxy)-3-oxopropyl)-23-oxo-2,5,8,11,14,17,20-heptaoxa-24-azahepta-cosan-27-oic acid
  • Example 10 24-(3-(((S)-1-(((S)-1-((4-((((4-(((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)oxy)methyl)-phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)-23-oxo-2,5,8,11,14,17,20-heptaoxa-24-azaheptacosan-27
  • RMN 1 H (400 MHz, ⁇ in ppm, DMSO-d6): 1.40 (s, 9H); 2.25 to 2.78 (m, 6H); 3.22 (s, 3H); 3.32 to 3.70 (m, 98H); 4.52 (m, 2H); 5.20 (m, 1H); 5.30 (m, 1H); 5.90 (m, 1H).
  • Compound 33 75-(3-(allyloxy)-3-oxopropyl)-74-oxo-2,5,8,11,14,17,20,23,26,29,32, 35,38,41,44,47,50,53,56,59,62,65,68,71-tetracosaoxa-75-azaoctaheptacontan-78-oic acid
  • Example 11 75-(3-(((S)-1-(((S)-1-((4-((((4-(((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)oxy)methyl)-phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)-74-oxo-2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,
  • example 11 16 mg, 7.1 ⁇ mol
  • DIEA 1.2 ⁇ L, 7.1 ⁇ mol
  • DSC 1.8 mg, 7.1 ⁇ mol
  • the reaction medium was stirred for 3 h at RT then concentrated in vacuo and purified by flash chromatography on 1.3 g of diol-modified silica gel (gradient elution DCM/iPrOH) to give 5 mg of a mixture of example 12 (30%) and example 11 (70%).
  • Compound 35 4-((3-(allyloxy)-3-oxopropyl)(3-(tert-butoxy)-3-oxopropyl)amino)-4-oxobutanoic acid
  • Compound 36 1-ammonio-15-oxo-3,6,9,12-tetraoxa-16-azaoctadecane-18-sulfonate
  • reaction medium was stirred at RT overnight, concentrated in vacuo and purified by preparative LCMS on a 5 ⁇ m C18 SunFire column (Waters, 30 ⁇ 100 mm, gradient elution MeCN+0.07% TFA/H 2 O+0.07% TFA) to give 341 mg of compound 36 (70%).
  • IR spectrum as a KBr pellet; main absorption bands in reciprocal centimeters: 3000; 1780; 1648; 1556; 1200-1170; 1200; 1100; 1040.
  • Compound 37 24-(3-(tert-butoxy)-3-oxopropyl)-4,20,23,27-tetraoxo-7,10,13,16,28-pentaoxa-3,19,24-triazahentriacont-30-ene-1-sulfonate
  • reaction medium was stirred for 2 h at RT, concentrated in vacuo and purified by preparative LCMS on a 5 ⁇ m C18 SunFire column (Waters, 30 ⁇ 100 mm, gradient elution MeCN+0.07% TFA/H 2 O+0.07% TFA) to give 118 mg of compound 37 (60%).
  • reaction medium was stirred for 2 h at RT, concentrated in vacuo and purified by reverse phase flash chromatography on 75 g of C18-modified silica gel (gradient elution H 2 O/MeCN) to give 60 mg of compound 39 as a white solid (24%).
  • Example 13 24-(3-(((S)-1-(((S)-1-((4-((((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)oxy)methyl)-phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)-4,20,23-trioxo-1-sulfo-7,10,13,16-tetraoxa-3,19,24-triazahept
  • the medium was concentrated in vacuo, then saturated aqueous NaHCO 3 (1 L) was added, followed by extraction with Et 2 O (2 ⁇ 250 mL), organic phases were excluded.
  • the aqueous phase was acidified with QS of aqueous 1N HCl to reach pH 2, and extracted with DCM (3 ⁇ 250 mL), the combined organic phases were dried over MgSO 4 , filtered and concentrated in vacuo to give 2.05 g compound 40 (85%).
  • reaction medium was stirred at RT for 24 h. At this time, piperidine (80 ⁇ L, 810 ⁇ mol) was added to the medium, and stirring was maintained for 1 h. Then, the reaction medium was concentrated in vacuo and purified by flash chromatography on 20 g silica gel (gradient elution DCM/MeOH) to give 60 mg of compound 43 (65%).
  • example 14 32 mg, 26 ⁇ mol in DMF (3 mL), under magnetic stirring, were added DSC (8 mg, 31.2 ⁇ mol) and DIEA (8.6 ⁇ L, 52 ⁇ mol). The reaction medium was stirred at RT overnight. Then, 4 mg of DSC were added and stirring at RT was maintained for 1 h. At this time, the reaction medium was diluted with MeTHF (10 mL), washed with H 2 O (5 mL). The aqueous phase was extracted with MeTHF (3 ⁇ 5 mL). The combined organic phases were dried over MgSO 4 , filtered and concentrated in vacuo. The medium was purified by flash chromatography on 4 g of silica gel (gradient elution DCM/iPrOH) to give 28 mg of example 15 (81%).
  • example 16 28.8 mg of hu2H11_R35-74 were reacted with 115.2 ⁇ L of a 10.05 mM solution of example 15 in DMA (6 eq.) for 2 h 30. After purification on Superdex 200 pg in buffer B pH 6.5+10% NMP, concentration on Amicon Ultra-15, dilution in buffer B pH 6.5 to a final concentration of NMP at 5% and filtration on 0.22 ⁇ m PVDF filter, 16.12 mg of ADC were obtained that contained 2.2% of residual example 14.
  • Compound 48 (S)-tert-butyl 2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)-5-oxo-5-((((2R,3S,4S,5R,6R)-3,4,5-trihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methyl)amino)pentanoate
  • reaction medium was stirred at RT overnight. Then, 25 mg of compound 49 and 25 ⁇ L of EDC were added to the medium, stirring was maintained for 1 h. At this time, piperidine (160 ⁇ L, 1.58 mmol) was added to the medium. After 2 h, the medium was concentrated in vacuo and purified by flash chromatography on 20 g of silica gel (gradient elution DCM/MeOH) to give 78 mg of compound 50 (45%).
  • Example 17 5-(((S)-1-(((S)-1-((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)amino)-1,5-dioxo-5-((((2R,3S,4S,5R,6R)-3,4,5-trihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methyl)amino)pentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoic acid
  • Example 17 (45 mg, 37 ⁇ mol) was diluted in toluene and concentrated in vacuo. Then, to a solution of example 17 in THF (5 mL), DCM (2 mL) and DMF (1 mL) were added, under magnetic stirring, DSC (11.4 mg, 44.4 ⁇ mol) and DIEA (19 ⁇ L, 112.6 ⁇ mol). The reaction medium was stirred at RT under Ar for 3 h. Then, DMF and DSC were added to the medium until completion of the reaction. After 3 h, the medium was diluted with water (5 mL), and extracted with MeTHF (3 ⁇ 10 mL). The combined organic phases were dried over MgSO 4 , filtered and concentrated in vacuo. The mixture was purified by flash chromatography on 4 g of silica gel (gradient elution DCM/iPrOH) to give 21 mg of example 18 (43%).
  • example 19 The general method described previously was used for the preparation of example 19. 29.02 mg of hu2H11_R35-74 were reacted with 116.6 ⁇ L of a 10 mM solution of example 18 in DMA (6 eq.) for 2 h 30. After purification on Superdex 200 pg in buffer B pH 6.5+10% NMP, concentration on Amicon Ultra-15, dilution in buffer B pH 6.5 to a final concentration of NMP at 5% and filtration on 0.22 ⁇ m PVDF filter, 23.1 mg of example 19 were obtained as a colorless limpid solution at a concentration of 2.2 mg/mL with a DAR of 2.8 (HRMS), a monomeric purity of 99.9% and a global yield of 78%.
  • HRMS DAR of 2.8
  • Free-drug level was above the threshold of 1%: the ADC was concentrated on Amicon Ultra-15, purified on Sephadex G25 in buffer B pH 6.5+5% NMP and filtrated on 0.22 ⁇ m PVDF filter to provide 17.25 mg of example 19 as a colorless limpid solution at a concentration of 1.50 mg/mL with a DAR of 2.55 (HRMS), a monomeric purity of 99.7% and a global yield of 59%.
  • HRMS 2.55
  • Compound 56 (S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)-5-(((2S,3R,4R,5R)-6-((tert-butyldiphenylsilyl)oxy)-2,3,4,5-tetrahydroxyhexyl)amino)-5-oxopentanoic acid
  • reaction medium was stirred for 3 h at RT then quenched with H 2 O (15 mL) and stirring was carried on 10 min.
  • the aqueous phase was extracted with DCM (3 ⁇ 20 mL), the combined organic phases were dried over MgSO 4 , filtered, concentrated in vacuo and purified by flash chromatography on 5 g of silica gel (gradient elution DCM/MeOH) to give 64 mg of Fmoc-protected dipeptide-cryptophycin intermediate as a white solid (64%).
  • Example 20 5-(((S)-1-(((S)-1-((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxy-benzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)amino)-1,5-dioxo-5-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-amino)pentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoic acid
  • reaction medium was stirred at RT overnight. Then, piperidine (32.8 ⁇ L, 327.4 ⁇ mol) was added to the medium, and stirring was maintained for 4 h 30. At this time, the medium was concentrated in vacuo, diluted with DMA and purified by flash chromatography on 15 g of C18-grafted silica gel (gradient elution H 2 O/CH 3 CN) to give 66 mg of compound 62(53%).
  • Example 21 (20S,23S)-20-((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)-23-isopropyl-14,22,25-trioxo-2,5,8,11-tetraoxa-15,21,24-triazanonacosan-29-oic acid
  • Example 22 (20S,23S)-2,5-dioxopyrrolidin-1-yl 20-((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)-23-isopropyl-14,22,25-trioxo-2,5,8,11-tetraoxa-15,21,24-triazanonacosan-29-oate
  • example 23 44.64 mg of hu2H11_R35-74 were reacted with 209.2 ⁇ L of a 10 mM solution of example 22 in DMA (7 eq.) for 3 h 30. After purification on Sephadex G25 in buffer B pH 6.5+5% NMP and concentration on Amicon Ultra-15, 36.6 mg of example 23 were obtained as a colorless limpid solution at a concentration of 1.83 mg/mL with a DAR of 3.6 (HRMS), a monomeric purity of 98.8% and a global yield of 82%.
  • HRMS DAR of 3.6
  • Free-drug level was above the threshold of 1%: the ADC was concentrated on Amicon Ultra-15, purified on Sephadex G25 in buffer B pH 6.5+5% NMP and filtrated on 0.22 ⁇ m PVDF filter to provide 32.6 mg of example 23 as a colorless limpid solution at a concentration of 1.63 mg/mL with a DAR of 3.3 (HRMS), a monomeric purity of 98.3% and a global yield of 73%.
  • HRMS DAR of 3.3
  • reaction medium was concentrated in vacuo, then diluted with water (200 mL), acidified with QS of aqueous 5N HCl, and extracted with DCM (2 ⁇ 200 mL). The combined organic phases were dried over MgSO 4 , filtered and concentrated in vacuo to afford 510 mg of compound 63 (53%).
  • reaction medium was stirred for 20 h at RT, partly concentrated in vacuo then diluted with H 2 O (10 mL), acidified to pH 3 with IR-120 (H) Amberlite resin (CAS number [78922-04-0]), filtered, concentrated in vacuo and purified by three consecutive flash chromatographies on 40 g, 25 g and 12 g of silica gel (gradient elution DCM/MeOH/H 2 O) to give 214 mg of compound 65 as a white lacquer (53%).
  • reaction medium was stirred for 3 h at RT before adding piperidine (91 ⁇ L, 915.3 ⁇ mol). Stirring was carried on for 1 h 30 at RT then the reaction medium was concentrated in vacuo and purified by two consecutive flash chromatographies on 25 g and 12 g of silica gel (gradient elution DCM/MeOH) to give 58 mg of compound 66 as a colorless lacquer (28%).
  • Example 24 (80S,83S)-80-((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)-83-isopropyl-74,82,85-trioxo-2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65, 68,71-tetracosaoxa-75,81,84-triazanonaoctacontan-89-oic acid
  • example 24 35 mg, 16.4 ⁇ mol
  • THF 3 mL
  • DSC 4.28 mg, 16.4 ⁇ mol
  • DIEA 2.7 ⁇ L, 16.4 ⁇ mol
  • the reaction medium was stirred for 20 h at RT, concentrated in vacuo and purified by flash chromatography on 4 g of silica gel (gradient elution DCM/iPrOH) to give 12.0 mg of example 25 as a white lacquer (33%).
  • example 26 After storage overnight at 4° C., purification on Superdex 200 pg in buffer B pH 6.5+20% NMP, concentration on Amicon Ultra-15, buffer exchange on Sephadex G25 in buffer B pH 6.5+5% NMP, concentration on Amicon Ultra-15 and filtration on 0.22 ⁇ m PVDF filter, 33.35 mg of example 26 were obtained as a colorless limpid solution at a concentration of 2.9 mg/mL with a DAR of 3.9 (HRMS), a monomeric purity of 100% and a global yield of 60%.
  • HRMS DAR of 3.9
  • reaction medium was stirred for 4 h at RT before adding piperidine (95 ⁇ L, 961.9 ⁇ mol); stirring was carried on for 2 h at RT then the reaction medium was concentrated in vacuo and purified by flash chromatography on 25 g of silica gel (gradient elution DCM/MeOH) to give 132 mg of compound 67 as a colorless oil (62%).
  • Example 27 (80S,83S)-80-((4-((2R,3R)-3-((S)-1-((3S,7S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-6,6,7-trimethyl-3-neopentyl-2,5,9,12-tetraoxo-1-oxa-4,8,11-triazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)-83-isopropyl-74,82,85-trioxo-2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65, 68,71-tetracosaoxa-75,81,84-triazanonaoctacontan-89-oic acid
  • example 27 To a solution of example 27 (33 mg, 15.2 ⁇ mol) in THF (3 mL) were added DSC (4 mg, 15.6 ⁇ mol) and DIEA (2.55 ⁇ L, 15.4 ⁇ mol). The reaction medium was stirred for 20 h at RT then were added DSC (1 mg, 3.9 ⁇ mol) and DIEA (1 ⁇ L, 6.0 ⁇ mol). The reaction medium was stirred for 2 h at RT, concentrated in vacuo and purified by flash chromatography on 4 g of silica gel (gradient elution DCM/iPrOH/MeCN) to give 11 mg of example 28 as a white lacquer (32%).
  • example 29 After storage overnight at 4° C., purification on Superdex 200 pg in buffer B pH 6.5+20% NMP, concentration on Amicon Ultra-15, buffer exchange on Sephadex G25 in buffer B pH 6.5+5% NMP and filtration on 0.22 ⁇ m PVDF filter, 30.1 mg of example 29 were obtained as a colorless limpid solution at a concentration of 2.51 mg/mL with a DAR of 4 (HRMS), a monomeric purity of 98.4% and a global yield of 67%.
  • HRMS DAR of 4
  • reaction medium was stirred at RT for 1 h. At this time, DMA (1 mL) was added, and stirring was maintained overnight. Then, EDC (20 ⁇ L) was added and the reaction medium was stirred for 2 more hours. Piperidine (12.9 ⁇ L, 129 ⁇ mol) was then added to the medium. After 1 h30 of stirring, the reaction medium was concentrated in vacuo and purified by 3 consecutive flash chromatographies on silica gel (isocratic elution 40:5:0.5 v/v/v DCM/MeOH/H 2 O and 12:3:0.5 v/v/v DCM/MeOH/NH 4 OH) to give 47 mg of compound 69 (33%).
  • example 30 (14 mg, 11.4 ⁇ mol), under magnetic stirring, were added DMF (3 mL), DSC (4.2 mg, 15.6 ⁇ mol) and DIEA (6 ⁇ L, 34.5 ⁇ mol).
  • the reaction medium was stirred at RT for 4 h.
  • the medium was diluted with MeTHF (10 mL) and washed with H 2 O (5 mL).
  • the aqueous phase was extracted with MeTHF (10 mL), the combined organic phases were dried over MgSO 4 , filtered concentrated in vacuo and purified by flash chromatography on 4 g of silica gel (gradient elution DCM/iPrOH) to give 3.5 mg of example 31 (23%).
  • example 32 After purification on Superdex 200 pg in buffer B pH 6.5+10% NMP, concentration on Amicon Ultra-15, dilution in buffer B pH 6.5 to a final concentration of NMP at 5% and filtration on 0.22 ⁇ m PVDF filter, 16.7 mg of example 32 were obtained as a colorless limpid solution at a concentration of 2.23 mg/mL with a DAR of 3 (HRMS), a monomeric purity of 100% and a global yield of 60%.
  • HRMS DAR of 3
  • Compound 70 di-tert-butyl 4,7,10,13-tetraoxahexadecane-1,16-dioate
  • IR spectrum as a KBr pellet; main absorption bands in reciprocal centimeters: 1458; 1344; 1212; 1184; 1046; 1037; 742; 737; 598; 576; 533 & 523.
  • reaction medium was stirred for 2 h at RT then acidified to pH 3 with Amberlite IR-120 (H) (CAS number [78922-04-0], filtered, concentrated in vacuo and purified by flash chromatography on 150 g of silica gel (DCM/MeOH/H 2 O) to give 824 mg of compound 74 as a colorless solid 61%).
  • Compound 75 sodium 1-carboxy-15-oxo-3,6,9,12-tetraoxa-16-azaoctadecane-18-sulfonate
  • reaction medium was stirred for 1 h at RT, acidified to pH 3 with Amberlit IR-120 (H) (CAS number [78922-04-0]), filtered, washed with DMF (5 mL), concentrated in vacuo and purified by flash chromatography on 100 g of silica gel (gradient elution DCM/MeOH/acetic acid) to give 447 mg of a colorless lacquer.
  • This lacquer was dissolved in 2:1 THF/H 2 O (120 mL), acidified to pH 3.3 with 0.1N HCl (about 4 mL) and concentrated in vacuo to give 354 mg of compound 76 as a white translucent solid (57%).
  • reaction medium as stirred for 2 h at RT then was added piperidine (134 ⁇ L, 1.36 mmol) and the stirring was carried on for 2 h.
  • the reaction medium was concentrated in vacuo and purified by two consecutive flash chromatographies on 50 g and 4 g of silica gel (gradient elution DCM/MeOH/H 2 O) to give 73 mg of compound 77 as a white solid (36%).
  • Example 33 sodium (25S,28S)-33-carboxy-25-((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclo-hexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)-28-isopropyl-4,19,27,30-tetraoxo-7,10,13,16-tetraoxa-3,20,26,29-tetraazatritriacontane-1-sulfonate
  • Example 34 sodium (25S,28S)-25-((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)-34-((2,5-dioxopyrrolidin-1-yl)oxy)-28-isopropyl-4,19,27,30,34-pentaoxo-7,10,13,16-tetraoxa-3,20,26,29-tetraazatetratriacontane-1-sulfonate
  • example 33 To a solution of example 33 (3.57 mg, 2.47 ⁇ mol) in DMA (93 ⁇ L) were added NHS (27.2 ⁇ L, 2.72 ⁇ mol) and EDC (111 ⁇ L, 2.22 ⁇ mol). The reaction medium was stirred for 48 h at RT: LCMS analysis showed that the presence of expected example 34 at 95% with 5% of residual example 33. This solution was used such as for conjugation and the preparation of example 35.
  • example 35 The general method described previously was used for the preparation of example 35. 11.16 mg of hu2H11_R35-74 were reacted with 53 ⁇ L of a 10 mM solution of example 34 in DMA (7 eq.) for 2 h 30 then were added 15 ⁇ L of a 10 mM solution of example 34 in DMA (2 eq.) for 2 h 30. After purification on PD-10 and Nap-10 columns in buffer B pH 6.5+5% NMP, 9.5 mg of example 34 were obtained as a colorless limpid solution at a concentration of 1.9 mg/mL with a DAR of 3.8 (HRMS), a monomeric purity of 97.5% and a global yield of 82%.
  • HRMS DAR of 3.8
  • Compound 80 4-((S)-20-((S)-2-amino-3-methylbutanamido)-14-oxo-2,5,8,11-tetraoxa-15-azahenicosanamido)benzyl 4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16yl)ethyl)-oxiran-2-yl)benzylcarbamate
  • reaction medium was stirred for 20 h at RT, concentrated in vacuo and purified by flash chromatography on 4 g of silica gel (gradient elution DCM/MeOH) to give 38 mg of Fmoc-protected intermediate as a colorless lacquer (92%).
  • Example 36 (20S,23S)-20-((4-((((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxy-benzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)oxy)methyl)-phenyl)carbamoyl)-23-isopropyl-14,22,25-trioxo-2,5,8,11-tetraoxa-15,21,24-triazanonacosan-29-oic acid
  • example 36 To a solution of example 36 (10 mg, 7.11 ⁇ mol) in THF (2 mL) were added DSC (2 mg, 7.65 ⁇ mol) and DIEA (1.2 ⁇ L, 7.26 ⁇ mol). The reaction medium was stirred for 20 h at RT, concentrated in vacuo and purified by two consecutive flash chromatographies on 0.6 g and 2 g of silica gel (gradient elution DCM/iPrOH) to give 3.3 mg of example 37 as a white lacquer (31%).
  • reaction medium was stirred for 20 h at RT then was added compound 82 (34 mg, 18.6 ⁇ mol) and stirring carried on for 4 h.
  • the reaction medium was concentrated in vacuo and purified by two consecutive flash chromatographies on 25 g of silica gel and 15 g of diol-modified silica gel (gradient elution DCM/MeOH) to give 152 mg of crude product that was purified by reverse phase chromatography on a 10 ⁇ m C18 column 250 ⁇ 40 mm (gradient elution MeCN/H 2 O) to give 78 mg of the Fmoc-protected intermediate containing 16% of compound 83 as a white powder (29%).
  • example 38 To a solution of example 38 (29 mg, 12.7 ⁇ mol) in THF (3 mL) were added DSC (3.6 mg, 13.8 ⁇ mol) and DIEA (2.1 ⁇ L, 12.9 ⁇ mol). The reaction medium was stirred for 20 h at RT, concentrated in vacuo and purified by flash chromatography on 4 g of silica gel (gradient elution DCM/iPrOH) to give 6.9 mg of example 39 as a colorless lacquer (23%).
  • the fractions containing the monomeric modified antibody were pooled and filtered through a Steriflip® filter unit (0.22 ⁇ m Durapore® PVDF membrane, Millipore) to provide 173 mg of modified antibody at a concentration of 5.75 mg/mL with a ratio of linker per antibody of 5.2 (HRMS), a monomeric purity of 97% and a global yield of 89%.
  • a Steriflip® filter unit (0.22 ⁇ m Durapore® PVDF membrane, Millipore
  • example 41 The general method described previously was used for the preparation of example 41. 45 mg of modified hu2H11_R35-74 were reacted with 604 ⁇ L of a 6.89 mM solution of example 40 in DMA (14 eq.) for 4 h 30 then were added 604 ⁇ L of a 10 mM solution of example 40 in DMA (14 eq.) and stirring was pursued overnight at RT.
  • example 41 After purification on Superdex 200 pg in buffer B pH 6.5+20% NMP, concentration on Amicon Ultra-15, gel filtration on PD-10 in buffer B pH 6.5+5% NMP and filtration on 0.22 ⁇ m PVDF filter, 28 mg of example 41 were obtained as a colorless limpid solution at a concentration of 1.6 mg/mL with a DAR of 3.44 (HRMS), a monomeric purity of 99.6% and a yield of 62%.
  • HRMS DAR of 3.44
  • Compound 85 sodium 2-(4-((3-(allyloxy)-3-oxopropyl)(3-(tert-butoxy)-3-oxopropyl)amino)-4-oxobutanamido)ethane-1-sulfonate
  • reaction medium was stirred at RT overnight then were added a solution of compound 73 (1.96 g, 13.3 mmol) in H 2 O (10 mL) and sodium hydrogenocarbonate (225.2 mg, 2.68 mmol).
  • the reaction medium was stirred for 1 h at RT, diluted with H 2 O (6 mL), acidified to pH 3 with Amberlite IR-120 (H) (CAS number [78922-04-4]), filtered, concentrated in vacuo and purified by flash chromatography on silica gel (gradient elution DCM/MeOH/H 2 O) to give 270 mg of compound 85 as a colorless foam (21%).
  • Example 42 sodium (20S,23S)-28-(2-carboxyethyl)-20-((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)-23-isopropyl-14,22,25,29,32-pentaoxo-2,5,8,11-tetraoxa-15,21,24,28,33-pentaazapentatriacontane-35-sulfonate
  • Example 43 sodium (6S,9S)-14-(2-carboxyethyl)-6-((4-((2R,3R)-3-((S)-1-((3S,10R,16S,E)-10-(3-chloro-4-methoxybenzyl)-3-isobutyl-6,6-dimethyl-2,5,9,12-tetraoxo-1,4-dioxa-8,11-diazacyclohexadec-13-en-16-yl)ethyl)oxiran-2-yl)benzyl)carbamoyl)-9-isopropyl-3,8,11,15,18-pentaoxo-1-((2R,3S,4S,5R,6R)-3,4,5-trihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)-2,7,10,14,19-pentaazahenicosane-21-sulfonate
  • reaction medium was stirred at RT overnight, diluted with DMF, filtered over 0.45 ⁇ m and purified by two consecutive reverse phase chromatographies on a 5 ⁇ m C18 column 30 ⁇ 150 mm (gradient elution MeCN/0.2N ammonium acetate pH 5.6) to give 4.7 mg of example 43 as a white solid (25%).
  • the conjugates of formula (V) were subjected to stability studies evaluating aggregation (looking at monomeric purity) and loss of DAR upon removal of the organic co-solvent used in the formulation and under accelerated stress conditions (10 day heating at 40° C.).
  • ADC stored in 10 mM phosphate 140 mM NaCl pH6.5 5% NMP were filtered on a SEC column to remove the NMP, formulated in 10 mM phosphate 70 mM NaCl 5% sucrose pH6.5 and analyzed by HRMS and SEC HPLC to determine the DAR and the monomeric purity.
  • ADC in 10 mM phosphate 70 mM NaCl 5% sucrose pH6.5 were heated at 40° C. during 10 days and analyzed by HRMS and SEC HPLC to determine the DAR and the monomeric purity.
  • Drug release by Cathepsin B was evaluated in vitro by incubating the ADC (1.5 ⁇ g/mL) in the presence of human Cathepsin B (Calbiochem #219362, 40 ⁇ g/mL) in acetate buffer pH 5 at 37° C. for 24 h and detecting the drug by UPLC-MS-SIR-DAD-ELSD. The slope was determined based on the kinetics over the 1 st hour, the half-life on the 24 h kinetics.
  • Cryptophycin conjugates of formula (V) were cleaved by human Cathepsin B with a kinetics similar to the one of reference ADCs.
  • MDA-MB-231 cells in their exponential growth phase were trypsinized and resuspended in their culture medium (DMEM/F12 Gibco #21331, 10% FCS Gibco #10500-056, 2 nM glutamine Gibco #25030).
  • the cell suspension was seeded in Cytostar 96-well culture plates (GE Healthcare Europe, #RPNQ0163) in the whole culture medium containing serum at a density of 5000 cells/well. After incubation for 4 h, successive dilutions of the ADC were added to the wells at concentrations decreasing from 10 ⁇ 7 to 10 ⁇ 12 M (in triplicate for each concentration).
  • the cells were cultured at 37° C. in an atmosphere containing 5% CO 2 in the presence of the ADC for 3 d.
  • Cryptophycin conjugates of formula (V), as well as ADC1 and ADC2 were found to inhibit the proliferation of MDA-MB-231 cell line with IC 50 ranging from 19 pM to 67 pM and selectivity ratio ADC alone vs ADC+naked antibody between 109 and >1493.
  • MTD was determined as the maximal dose that does not induce 15% body weight loss during 3 consecutive days for an individual mouse or 20% body weight loss during 1 day or mortality. It was evaluated after a single intravenous (i.v.) bolus injection in 3 female SCID mice and during a period of 28 days post-treatment.
  • Tested cryptophycin conjugates of formula (V) displayed MTD in SCID mice ranging from 30 mg/kg to >40 mg/kg.
  • the primary efficacy end points were ⁇ T/ ⁇ C, percent median regression, partial and complete regressions (PR and CR).
  • Changes in tumor volume for each treated (T) and control (C) were calculated for each tumor by subtracting the tumor volume on the day of first treatment (staging day) from the tumor volume on the specified observation day.
  • Tumor free survivor (TFS) was defined as the animals with undetectable tumors at the end of the study (>100 days post last treatment).
  • the invention also relates to the use of cryptophycin conjugates of formula (V) as anticancer agents.
  • the invention also relates to the use of cryptophycin conjugates of formula (V) for the preparation of a medicament for treating cancer, for instance breast cancer.
  • the present invention also provides conjugates of formula (V) according to the present invention for use in the treatment of cancer.
  • the present invention also provides medicaments which comprise at least one conjugate of formula (V).
  • These medicaments are employed therapeutically, especially in the treatment of cancer, for instance breast cancer.
  • the present invention relates to pharmaceutical compositions comprising as active principle a conjugate of formula (V) according to the invention.
  • These pharmaceutical compositions comprise an effective dose of at least one conjugate of formula (V) according to the invention and also at least one pharmaceutically acceptable excipient.
  • the present invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising at least one conjugate of formula (V) according to the present invention, and also at least one pharmaceutically acceptable excipient.
  • excipients are selected, in accordance with the pharmaceutical form and method of administration desired, from the customary excipients, which are known to a person skilled in the art.
  • the present invention also provides a method of treating the pathologies indicated above such as cancer, for instance breast cancer, which comprises administering to a patient an effective dose of a conjugate of formula (V) according to the invention.

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US11951165B2 (en) 2016-12-30 2024-04-09 Vaxcyte, Inc. Conjugated vaccine carrier proteins
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CZ64599A3 (cs) * 1996-08-30 1999-10-13 Eli Lilly And Company Farmaceutické sloučeniny
US20050175619A1 (en) 2004-02-05 2005-08-11 Robert Duffy Methods of producing antibody conjugates
USRE47123E1 (en) 2006-07-18 2018-11-13 Sanofi EPHA2 receptor antagonist antibodies
FR2947269B1 (fr) 2009-06-29 2013-01-18 Sanofi Aventis Nouveaux composes anticancereux
KR101364475B1 (ko) 2009-06-30 2014-02-19 알까뗄 루슨트 무선 로컬 영역 네트워크, 관련된 액세스 제어기 및 액세스 포인트 디바이스에서 모바일 단말을 위한 로밍 방법
AR078470A1 (es) 2009-10-02 2011-11-09 Sanofi Aventis Anticuerpos que se unen especificamente al receptor epha2
CA2934030A1 (en) * 2013-10-15 2015-04-23 Sorrento Therapeutics Inc. Drug-conjugates with a targeting molecule and two different drugs
MX2021008464A (es) 2013-10-15 2023-03-03 Seattle Genetics Inc Farmaco-enlazadores modificados con polietilenglicol para la farmacocinetica mejorada de conjugados de ligando-farmaco.
WO2015073575A2 (en) * 2013-11-12 2015-05-21 Centre For Probe Development And Commercialization Residualizing linkers and uses thereof
WO2016065145A2 (en) 2014-10-22 2016-04-28 The Johns Hopkins University Psma targeted reversed carbamates and methods of use thereof
EP3229844B1 (en) * 2014-12-09 2020-03-04 AbbVie Inc. Antibody drug conjugates with cell permeable bcl-xl inhibitors
JP2018515457A (ja) * 2015-04-21 2018-06-14 アッヴィ・ステムセントルクス・エル・エル・シー カリケアマイシン構築物および使用方法
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CN108066772B (zh) * 2016-11-14 2021-07-13 中国科学院上海药物研究所 靶向tacstd2的抗体与药物偶联体(adc)分子

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