WO2019042447A1 - 一种新毒素及其中间体的制备方法 - Google Patents

一种新毒素及其中间体的制备方法 Download PDF

Info

Publication number
WO2019042447A1
WO2019042447A1 PCT/CN2018/103781 CN2018103781W WO2019042447A1 WO 2019042447 A1 WO2019042447 A1 WO 2019042447A1 CN 2018103781 W CN2018103781 W CN 2018103781W WO 2019042447 A1 WO2019042447 A1 WO 2019042447A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
formula
group
antibody
protecting group
Prior art date
Application number
PCT/CN2018/103781
Other languages
English (en)
French (fr)
Inventor
许建烟
章瑛
屈博磊
蒋贵阳
Original Assignee
江苏恒瑞医药股份有限公司
上海恒瑞医药有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 江苏恒瑞医药股份有限公司, 上海恒瑞医药有限公司 filed Critical 江苏恒瑞医药股份有限公司
Priority to CN201880004457.6A priority Critical patent/CN109963835B/zh
Publication of WO2019042447A1 publication Critical patent/WO2019042447A1/zh

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/44—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members
    • C07D207/444—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members having two doubly-bound oxygen atoms directly attached in positions 2 and 5
    • C07D207/448—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members having two doubly-bound oxygen atoms directly attached in positions 2 and 5 with only hydrogen atoms or radicals containing only hydrogen and carbon atoms directly attached to other ring carbon atoms, e.g. maleimide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the invention relates to a method for preparing a new toxin and an intermediate thereof.
  • Patent application WO2016127790A1 (Publication No. 2016.08.18) relates to a class of ligand-cytotoxic drug conjugates, which are regulated by a receptor in the preparation of a medicament for treating cancer, by a receptor-cytotoxic drug conjugate
  • the compound represented by the formula (I) is found to be an important intermediate, and the structure is as follows:
  • the preparation method is as follows:
  • the compound 1i is directly condensed with the compound 7d to give the intermediate 7e, and finally reacted with 6-maleimidocaproyl chloride to obtain the target compound Mc-MMAF analog, which is a total of four steps.
  • Mc-MMAF The preparation method of Mc-MMAF is disclosed in the patent application WO2005081711A2 (publication date 2005.09.09).
  • the synthesis method of the invention adopts two fragments separately synthesized, and finally directly condenses to obtain the compound represented by the general formula (I), and the total yield is greatly improved.
  • the synthetic route of the method has the characteristics of low synthesis cost and is suitable for industrial production, and has remarkable characteristics. Social and economic benefits.
  • the technical problem to be solved by the present invention is to provide a process for preparing a compound of the formula (I) and an intermediate thereof, which comprises a compound of the formula (III) which undergoes a series of upper protecting groups, deprotecting groups, Amidation reaction with 6-maleimidocaproyl chloride to obtain a compound of the formula (I), which overcomes the limitations of the prior art on the substrate, has mild reaction conditions, is easy to operate, has high optical purity and is synthesized.
  • the advantages of low cost and the like are suitable for mass production.
  • the present invention provides a compound of the formula (III):
  • R 1 is selected from a hydrogen atom or a carboxyl protecting group.
  • the carboxy protecting group is selected from the group consisting of methyl, substituted methyl, ethyl, 2-substituted ethyl, allyl, tert-butyl, alkoxyalkyl, alkoxyalkoxyalkyl a class, a 2,6-dialkylphenyl group, a benzyl group, a substituted benzyl group, a silane group or a tin alkyl group;
  • the substituted methyl group is selected from the group consisting of 9-fluorenylmethyl, triisopropylsilylmethyl, cyclopropylmethyl, diphenylmethyl or triphenylmethyl;
  • the 2-substituted ethyl group is selected from the group consisting of 2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(p-toluenesulfonyl)ethyl or 2-cyanoethyl base;
  • the alkoxyalkyl group is selected from the group consisting of methoxymethyl, benzyloxymethyl or triisopropylsiloxymethyl;
  • the alkoxyalkoxyalkyl group is selected from the group consisting of methoxyethoxymethyl groups
  • the 2,6-dialkylphenyl group is selected from the group consisting of 2,6-dimethylphenyl, 2,6-diisopropylphenyl or 2,6-di-tert-butyl-4-methoxybenzene base;
  • the substituted benzyl group is selected from the group consisting of p-methylbenzyl, 2,4-dimethoxybenzyl, 2,6-dimethoxybenzyl, p-nitrobenzyl or o-nitrobenzyl;
  • the silane group is selected from the group consisting of trimethylsilyl, triethylsilyl, triisopropylsilyl or phenyldimethylsilyl;
  • the tin alkyl group is selected from the group consisting of trimethylstannyl
  • the compound of the formula (III) is selected from the group consisting of:
  • the invention further relates to a process for the preparation of a compound of the formula (III), which comprises reacting a compound of the formula (II) with 6-maleimidocaproic acid to give a formula (III) The step of showing the compound,
  • R 1 is as defined in formula (III);
  • R 2 is selected from a hydrogen atom or an amino protecting group
  • the amino protecting group is preferably an alkoxycarbonylamino protecting group, an acylamino protecting group, a sulfonylamino protecting group or an alkylamino protecting group;
  • the alkoxycarbonylamino protecting group is selected from the group consisting of benzyloxycarbonyl, tert-butoxycarbonyl, fluorenylmethoxycarbonyl, allyloxycarbonyl, trimethylsilyloxycarbonyl, methoxycarbonyl or ethoxycarbonyl;
  • the acylamino protecting group is selected from the group consisting of phthaloyl, trifluoroacetyl, pivaloyl, benzoyl, formyl or acetyl;
  • the sulfonyl amino protecting group is selected from p-toluenesulfonyl, o-nitrophenylsulfonyl or p-nitrophenylsulfonyl;
  • the alkyl-based amino protecting group is selected from the group consisting of trityl, 2,4-dimethoxybenzyl, p-methoxybenzyl or benzyl; preferably an alkoxycarbonyl-based amino protecting group, more preferably t-butoxy Carbonyl or fluorenylmethoxycarbonyl.
  • the reaction is a condensation reaction
  • the amide bond-forming condensing agent is selected from the group consisting of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-dicyclohexylcarbamate Imine, N,N'-diisopropylcarbodiimide, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate, 1-hydroxybenzene And triazole, 1-hydroxy-7-azobenzotriazole, O-benzotriazole-N,N,N',N'-tetramethyluron hexafluorophosphate, 2-(7-even Nitrobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphat
  • the above condensation reaction is preferably carried out under basic conditions selected from the group consisting of organic or inorganic bases, preferably triethylamine, diethylamine, N,N-diisopropylethylamine (diisopropyl) Ethyl ethylamine), pyridine, sodium hexamethyldisilazide, n-butyllithium, potassium t-butoxide or tetrabutylammonium bromide, the inorganic base being selected from the group consisting of lithium hydroxide, sodium hydroxide, and hydroxide Potassium, sodium hydride, sodium carbonate, sodium hydrogencarbonate, potassium carbonate, potassium hydrogencarbonate or cesium carbonate, more preferably N,N-diisopropylethylamine.
  • organic or inorganic bases preferably triethylamine, diethylamine, N,N-diisopropylethylamine (diisopropyl) Ethyl
  • R 1 carboxy protecting group is DMB (2,4-dimethoxybenzyl)
  • the compound of the formula (II) is as shown in the compound of the formula (IIa).
  • the compound of the formula (III) is represented by the compound of the formula (IIIa); or when the R 1 is a hydrogen atom, the compound of the formula (III) is represented by the compound of the formula (IIIb).
  • the method is
  • the compound of the formula (IIa) is condensed with 6-maleimidocaproic acid to give a compound of the formula (IIIa).
  • the method further comprises the following steps
  • the compound of the formula (II), wherein R 1 is a hydrogen atom, R 2 is an amino protecting group, the compound of the formula (II) is as shown by the compound of the formula (IIc); more preferably, The R 1 carboxy protecting group is DMB (2,4-dimethoxybenzyl), R 2 is an amino protecting group, and the compound of formula (II) is as shown in the compound of formula (IIb).
  • the method further comprises the following steps
  • the method further comprises the compound of the formula (IIc), wherein a compound represented by the formula (IIb) is obtained by adding a carboxy protecting group, and the compound of the formula (IIb) is deprotected by a Fmoc protecting group under basic conditions.
  • the base is selected from an organic base or an inorganic base, and the organic base is preferably triethylamine, diethylamine, N,N'-diisopropylethylamine, pyridine, sodium hexamethyldisilazide, n-butyl a lithium base, potassium t-butoxide or tetrabutylammonium bromide, the inorganic base being selected from the group consisting of lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium hydride, sodium carbonate, sodium hydrogencarbonate, potassium carbonate, potassium hydrogencarbonate or Barium carbonate, more preferably diethylamine.
  • the present invention also relates to a process for the preparation of a compound of the formula (I), which comprises reacting a compound of the formula (III) with a compound of the formula (IV) to give a compound of the formula (V): a step of deprotecting a compound of (V) to give a compound of formula (I),
  • A is selected from a 3-8 membered cycloalkyl group, preferably a 3-membered cycloalkyl group;
  • R is selected from halogen, preferably fluorine, chlorine, bromine or iodine, most preferably fluorine;
  • R 1 is as defined in formula (III);
  • R 3 is selected from a carboxy protecting group selected from the group consisting of 2,4-dimethoxybenzyl, benzyl, allyl, pentafluorophenyl, methyl, p-methylbenzyl, methoxy Ethoxymethyl or tert-butyl, preferably 2,4-dimethoxybenzyl;
  • R x is selected from an optionally substituted alkyl or cycloalkyl group selected from alkyl, haloalkyl, halogen, hydroxy, amino, nitro, cyano, alkoxy, hydroxyalkyl, cycloalkyl a heterocyclic group, an aryl group or a heteroaryl group;
  • n is selected from 1, 2, 3, 4 or 5.
  • the method further comprises the step of: condensing a compound of the formula (III) with a compound of the formula (IV) by a compound of the formula (III) to obtain a compound of the formula (V),
  • the condensing agent is selected from the group consisting of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide, N,N'-diiso Propylcarbodiimide, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7- Azobenzotriazole, O-benzotriazole-N,N,N',N'-tetramethyluron hexafluorophosphate, 2-(7-azobenzotriazole)-N, N,N',N'-tetramethylurea hexafluorophosphate, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate or benzotriazole hexafluorophosphate-
  • the above condensation reaction is preferably carried out under basic conditions selected from the group consisting of organic or inorganic bases, preferably triethylamine, diethylamine, N,N-diisopropylethylamine (diisopropyl) Ethyl ethylamine), pyridine, sodium hexamethyldisilazide, n-butyllithium, potassium t-butoxide or tetrabutylammonium bromide, the inorganic base being selected from the group consisting of lithium hydroxide, sodium hydroxide, and hydroxide Potassium, sodium hydride, sodium carbonate, sodium hydrogencarbonate, potassium carbonate, potassium hydrogencarbonate or cesium carbonate, more preferably N,N-diisopropylethylamine.
  • organic or inorganic bases preferably triethylamine, diethylamine, N,N-diisopropylethylamine (diisopropyl) Ethyl
  • the method is
  • the method comprises the condensation of a compound of the formula (IIIb) with a compound of the formula (IVa) under basic conditions and a condensing agent to obtain a compound of the formula (Va), (Va).
  • the compound is deprotected under the action of an acid to give a compound of the formula (Ia).
  • the condensing agent is selected from the group consisting of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide, N,N'-diiso Propylcarbodiimide, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7- Azobenzotriazole, O-benzotriazole-N,N,N',N'-tetramethyluron hexafluorophosphate, 2-(7-azobenzotriazole)-N, N,N',N'-tetramethylurea hexafluorophosphate, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate or benzotriazole hexafluorophosphate
  • the base is selected from an organic base or an inorganic base, and the organic base is preferably triethylamine, diethylamine, N,N-diisopropylethylamine (diisopropylethylamine), pyridine or hexamethyl.
  • Sodium silylamino, n-butyllithium, potassium t-butoxide or tetrabutylammonium bromide the inorganic base being selected from the group consisting of lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium hydride, sodium carbonate, sodium hydrogencarbonate Potassium carbonate, potassium hydrogencarbonate or cesium carbonate, more preferably N,N-diisopropylethylamine.
  • Agents that provide acidic conditions include, but are not limited to, hydrogen chloride, hydrogen chloride in 1,4-dioxane solution, ammonium chloride, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid, phosphoric acid, and para-benzene. Methanesulfonic acid, preferably trifluoroacetic acid.
  • the invention also provides a process for the preparation of a compound of formula (IVa):
  • the compound of the formula (M3) is amidated with a compound of the formula (i) by a condensing agent to obtain a compound of the formula (IVb), and the compound of the formula (IVb) is deprotected by a base. a compound of the formula (IVa),
  • the condensing agent is selected from the group consisting of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide, N,N'-diiso Propylcarbodiimide, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7- Azobenzotriazole, O-benzotriazole-N,N,N',N'-tetramethyluron hexafluorophosphate, 2-(7-azobenzotriazole)-N, N,N',N'-tetramethylurea hexafluorophosphate, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate or benzotriazole hexafluorophosphate-
  • the base is selected from an inorganic base or an organic base, preferably an organic base selected from the group consisting of amine organic bases such as diethylamine, triethylamine or aqueous ammonia.
  • the invention also provides a preparation method of the compound of the formula (M3):
  • the base is selected from an inorganic base or an organic base, preferably an organic base selected from the group consisting of amine organic bases such as diethylamine, triethylamine, 4-dimethylaminopyridine or 1-(3-dimethyl Aminopropyl)-3-ethylcarbodiimide.
  • amine organic bases such as diethylamine, triethylamine, 4-dimethylaminopyridine or 1-(3-dimethyl Aminopropyl)-3-ethylcarbodiimide.
  • the invention further provides a process for the preparation of a compound of formula (i):
  • the compound of the formula (a) is reacted with a compound of the formula (b) under the action of an organic base and an organic boron to give a compound of the formula (c), the compound of the formula (c) in an oxidizing agent (such as hydrogen peroxide) and a base.
  • the reaction is carried out under the action of lithium hydroxide monohydrate to obtain a compound of the formula (d).
  • the compound of the formula (d) is reacted with a benzyl bromide in the presence of a base such as sodium hydrogencarbonate to obtain a compound of the formula (e).
  • the compound of the formula (e) is reacted with methyl iodide to obtain a compound of the formula (f), and the compound of the formula (f) is deprotected to give a compound of the formula (g), and the compound of the formula (g) is reacted with hydrochloric acid to obtain a compound of the formula (h) wherein a compound of the formula (h) is reacted with a 9-fluorenylmethyl-N-succinimidylcarboxylate under the action of a base such as N,N-diisopropylethylamine. a compound of formula (i).
  • the present invention also provides a method for producing an antibody drug conjugate or a pharmaceutically acceptable salt thereof, wherein the antibody drug conjugate is passed from the antibody to the linker unit and the formula (I) in the above scheme
  • the compound is preferably conjugated, and the antibody is preferably a TNF- ⁇ antibody, a VEGF antibody, a c-MET antibody, an EGFR antibody, a Her2 antibody or a PD-1 antibody, and the linker unit is preferably an alkylene group, an arylene group or a heteroarylene group. , alkylene sulfanyl or iminothioalkyl.
  • the TNF- ⁇ antibody is selected from the group consisting of etanercept, infliximab, or adalimumab
  • the VEGF antibody is selected from bevacizumab
  • the EGFR antibody is selected from the group consisting of cetuximab, Pani Monoclonal antibody or nimotuzumab
  • the Her2 antibody is selected from trastuzumab (Herceptin) or pertuzumab
  • the PD-1 antibody is selected from the group consisting of navobizumab or palmeduzumab.
  • the antibody drug conjugate is selected from the group consisting of
  • the antibody Ab is preferably a TNF- ⁇ antibody, a VEGF antibody, an EGFR antibody, a Her2 antibody or a PD-1 antibody, and y is in the range of 1-8, preferably 2-5; y is a positive real number and may be an integer or a non-integer.
  • halogenated as used in the present invention means substituted by "halogen atom", and "halogen atom” means a fluorine atom, a chlorine atom, a bromine atom, an iodine atom or the like.
  • alkyl group in the present invention means a linear or branched alkyl group having 1 to 6 carbon atoms, and includes, for example, "C 1-4 alkyl group", "C 1-3 alkyl group” and the like, and specific examples include But not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, new Pentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2 - dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-B A butyl group
  • haloalkyl refers to a group derived by substituting one or more "halogen atoms” for one or more hydrogen atoms on an "alkyl group” as described above. Defined.
  • hydroxyalkyl refers to a radical derived from one or more "hydroxy” groups substituted by one or more hydrogen atoms on an "alkyl group” as defined above.
  • alkoxy group as used in the present invention means a group which is bonded in an alkyl-O- form, wherein “alkyl group” is as defined above.
  • cycloalkyl group as used in the present invention means a saturated or partially unsaturated monocyclic or polycyclic cyclic group, and the number of ring atoms includes 3 to 20, preferably 3 to 8 carbon atoms, more preferably 3 to 6 carbon atom. Specific examples include, but are not limited to, cyclopropyl, cyclobutane, cyclopentyl, cyclohexane, cycloheptyl, cyclooctyl and the like.
  • heterocyclic group as used in the present invention means a cyclic group having 3 to 8 ring atoms in which at least one ring atom is a hetero atom such as a nitrogen atom, an oxygen atom or a sulfur atom.
  • a ring atom in the cyclic structure may be oxidized, preferably a "5-6 membered heterocyclic group.”
  • a ring atom e.g., a carbon atom, a nitrogen atom, or a sulfur atom
  • a ring atom in the cyclic structure may be oxidized, preferably a "5-6 membered heterocyclic group.”
  • Specific examples include, but are not limited to, aziridine, 2H-azepine, diaziryl, 3H-diazapropenyl, azetidinyl, 1,4-dioxo Heterocyclohexane, 1,3-dioxanyl, 1,3-dioxolyl, 1,4-dioxadienyl, tetrahydrofuranyl, dihydropyrrolyl , pyrrolidinyl, imidazolidinyl, 4,5-dihydroimidazolyl, pyrazolidin
  • aryl group as used in the present invention means an aromatic cyclic group having 6 to 15 ring carbon atoms, and includes “6-8 membered aryl group” and "6-15 membered fused aryl group".
  • a ring atom for example, a carbon atom, a nitrogen atom or a sulfur atom
  • examples thereof include, but are not limited to, phenyl, cyclooctadecenyl, naphthyl, anthryl, phenanthryl and the like.
  • heteroaryl group as used in the present invention means an aromatic cyclic group having 5 to 15 ring atoms (at least one of which is a hetero atom such as a nitrogen atom, an oxygen atom or a sulfur atom), including "5-8 membered heteroaryl", “5-15 membered heteroaryl”, optionally, a ring atom (for example, a carbon atom, a nitrogen atom or a sulfur atom) in the cyclic structure may be oxidized.
  • a "linker unit” as used herein refers to a chemical moiety comprising a covalent bond or a chain of atoms that covalently attaches an antibody to a drug moiety, for example, an alkylene group, an arylene group, a heteroarylene group, an alkylene sulfanyl group or The iminothioalkyl group, the linker unit together with the maleimide group of formula (I), forms a complete linker.
  • Linkers are known in the art, such as SMCC, SPP, SPDP, and the like.
  • Substituted as used in the present invention means that one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3, hydrogen atoms are independently substituted with each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art will be able to determine (by experiment or theory) substitutions that may or may not be possible without undue effort. For example, an amino group or a hydroxyl group having a free hydrogen may be unstable when combined with a carbon atom having an unsaturated (e.g., olefinic) bond.
  • the reaction step is shortened, and the reactants in the key step of the preparation of the compound of the formula (I) are different in the present invention.
  • the first step in the key step of the present invention consists of the compound of the formula (IIIb) and the formula (IVa).
  • the compound shown is reacted to give a compound of the formula (Va), and the second step is obtained by decarboxylation of a compound represented by the formula (Va) to give a compound of the formula (Ia), which is compared with the prior art.
  • the structure of the compound is determined by nuclear magnetic resonance (NMR) or/and mass spectrometry (MS).
  • NMR shift ( ⁇ ) is given in units of 10-6 (ppm).
  • the NMR was measured by a Bruker AVANCE-400 nuclear magnetic apparatus, and the solvent was deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), and the internal standard was tetramethylsilane ( TMS).
  • DMSO-d6 dimethyl sulfoxide
  • CDCl3 deuterated chloroform
  • CD3OD deuterated methanol
  • TMS tetramethylsilane
  • the measurement of the MS was carried out using a FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Thermo, model: Finnigan LCQ advantage MAX).
  • ESI FINNIGAN LCQAd
  • the HPLC was measured using an Agilent 1200 DAD high pressure liquid chromatograph (Sunfire C18 150 x 4.6 mm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18 150 x 4.6 mm column).
  • Chiral HPLC analysis assays were performed using LC-10A vp (Shimadzu) or SFC-analytical (Berger Instruments Inc.).
  • the average inhibition rate of the kinase and the IC50 value were determined using a NovoStar plate reader (BMG, Germany).
  • Thin layer chromatography silica gel plate uses Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate.
  • the specification of silica gel plate used for thin layer chromatography (TLC) is 0.15mm ⁇ 0.2mm.
  • the specification for thin layer chromatography separation and purification is 0.4mm. ⁇ 0.5mm.
  • the known starting materials of the present invention may be synthesized by or according to methods known in the art, or may be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Dari Companies such as chemicals.
  • the reactions can all be carried out under an argon atmosphere or a nitrogen atmosphere.
  • An argon atmosphere or a nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon having a volume of about 1 L.
  • the hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon of about 1 L volume.
  • the pressurized hydrogenation reaction was carried out using a Parr Model 3916EKX hydrogenation apparatus and a clear blue QL-500 type hydrogen generator or a HC2-SS type hydrogenation apparatus.
  • the hydrogenation reaction is usually evacuated, charged with hydrogen, and operated three times.
  • the microwave reaction used a CEM Discover-S Model 908860 microwave reactor.
  • the solution means an aqueous solution.
  • reaction temperature is room temperature and is 20 ° C to 30 ° C.
  • the progress of the reaction in the examples was monitored by thin layer chromatography (TLC).
  • TLC thin layer chromatography
  • the system used for the reaction was: A: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: petroleum ether And the ethyl acetate system, D: acetone, the volume ratio of the solvent is adjusted depending on the polarity of the compound.
  • Purification compounds using column chromatography eluent systems and thin layer chromatography developer systems include: A: dichloromethane and methanol systems, B: n-hexane and ethyl acetate systems, C: dichloromethane and acetone
  • A dichloromethane and methanol systems
  • B n-hexane and ethyl acetate systems
  • C dichloromethane and acetone
  • the volume ratio of the solvent is adjusted depending on the polarity of the compound, and a small amount of an alkaline or acidic reagent such as triethylamine or acetic acid may be added for adjustment.
  • the reaction mixture was evaporated to dryness and evaporated, evaporated, evaporated, evaporated
  • the organic layer was dried (MgSO4), filtered, evaporated,
  • the aqueous phase was washed with dichloromethane (150 mL ⁇ 3), and a 2N hydrochloric acid solution was added dropwise to pH 2-3 under ice bath, extracted with ethyl acetate (100 mL ⁇ 3), washed with water (500 mL) The sodium solution (500 mL) was evaporated.
  • the amount of the intermediate 1n required is completely different, the amount of the intermediate 1n of the present invention can be greatly reduced, the amount of the intermediate 1n is 1.94 g, and the prior art is 2.82 g; the present invention and the present invention
  • the technically important reaction steps are different.
  • the present invention is a 2-step reaction, and the prior art is a 4-step reaction.
  • the prior art requires a multi-step complex route synthesis of the compound of Formula 1, which can be greatly reduced after the synthesis method of the present invention is employed. Production costs, shortened reaction time, improved production efficiency, outstanding technical progress and significant economic effects.

Abstract

一种新毒素及其中间体的制备方法，具体包括通式（III）化合物及其制备方法，以及通式（III）化合物经过一系列的上保护基、脱保护基和缩合反应得到通式（I）化合物的制备方法。

Description

一种新毒素及其中间体的制备方法 技术领域
本发明涉及一种新毒素及其中间体的制备方法。
背景技术
专利申请WO2016127790A1(公开日2016.08.18)中涉及一类配体-细胞毒性药物偶联物，该配体-细胞毒性药物偶联物及其药物组合物通过受体调节在制备治疗癌症的药物中的用途中具有良好的效果，在合成该类配体-细胞毒性药物偶联物时，发现通式(I)所示化合物是重要的中间体，结构如下所示：
Figure PCTCN2018103781-appb-000001
其制备方法如下所示：
Figure PCTCN2018103781-appb-000002
在该合成路线中，采用化合物1i与化合物7d直接缩合得到中间体7e，最后与6-马来酰亚氨基己酰氯反应得到目标化合物Mc-MMAF类似物，总计四步反应。
专利申请WO2005081711A2(公开日2005.09.09)中公开了Mc-MMAF的制备方法。
在合成本发明化合物的实验过程中，采用上述类似方法合成本发明通式(I)所 示化合物的过程中，总路线较长，产率较低，因此利用现有的合成方法大量合成通式(I)所示化合物还存在不少困难和不足，有必要开发高产率并适用于大规模合成通式(I)所示化合物的方法。
本发明的合成方法，采用两个片段分别合成，最后直接缩合得到通式(I)所示的化合物，总产率大大提高，该方法合成路线具有合成成本低，适合工业化生产等特点，具有显著的社会效益和经济效益。
本申请人的专利申请WO2017144015A1中也公开了(I)所示化合物的制备方法，其发明思路与本发明完全不同。
发明内容
本发明要解决的技术问题是提供一种制备式(I)所示化合物及其中间体的方法，所述方法包含通式(III)所示化合物经过一系列的上保护基、脱保护基、与6-马来酰亚氨基己酰氯发生酰胺化反应得到通式(I)所示的化合物，该方法克服现有技术对底物的限制，具有反应条件温和，操作简便，光学纯度高和合成成本低廉等优点，适于大规模生产。
本发明的技术方案如下：
本发明提供一种式(III)所示化合物：
Figure PCTCN2018103781-appb-000003
其中：
R 1选自氢原子或羧基保护基，
所述羧基保护基选自甲基、取代的甲基类、乙基、2-取代的乙基类、烯丙基、叔丁基、烷氧基烷基类、烷氧基烷氧基烷基类、2,6-二烷基苯基类、苄基、取代的苄基类、硅烷基类或锡烷基类；
所述取代的甲基类选自9-芴基甲基、三异丙基硅甲基、环丙基甲基、二苯基甲基或三苯基甲基；
所述2-取代的乙基类选自2,2,2-三氯乙基、2-(三甲基硅烷基)乙基、2-(对甲苯磺酰基)乙基或2-氰基乙基；
所述烷氧基烷基类选自甲氧基甲基、苄氧基甲基或三异丙基硅氧基甲基；
所述烷氧基烷氧基烷基类选自甲氧基乙氧基甲基；
所述2,6-二烷基苯基类选自2,6-二甲基苯基、2,6-二异丙基苯基或2,6-二叔丁基-4-甲氧基苯基；
所述取代的苄基类选自对甲基苄基、2,4-二甲氧基苄基、2,6-二甲氧基苄基、对硝基苄基或邻硝基苄基；
所述硅烷基类选自三甲基甲硅烷基、三乙基甲硅烷基、三异丙基甲硅烷基或苯基二甲基甲硅烷基、；
所述锡烷基类选自三甲基甲锡烷基；
优选甲基、烯丙基、叔丁基、苄基、2,4-二甲氧基苄基、对甲基苄基、五氟代苯基或甲氧乙氧甲基。
优选的，通式(III)所示化合物选自：
Figure PCTCN2018103781-appb-000004
本发明进一步涉及一种式(III)所示化合物的制备方法，其特征在于，所述方法包括由式(II)所示化合物与6-马来酰亚氨基己酸反应得到式(III)所示化合物的步骤，
Figure PCTCN2018103781-appb-000005
其中，
R 1如式(III)中所定义；
R 2选自氢原子或氨基保护基；
所述氨基保护基优选烷氧羰基类氨基保护基、酰基类氨基保护基、磺酰基类氨基保护基或烷基类氨基保护基；
所述烷氧羰基类氨基保护基选自苄氧羰基、叔丁氧羰基、笏甲氧羰基、烯丙氧羰基、三甲基硅乙氧羰基、甲氧羰基或乙氧羰基；
所述酰基类氨基保护基选自邻苯二甲酰基、三氟乙酰基、特戊酰基、苯甲酰基、甲酰基或乙酰基；
所述磺酰基类氨基保护基选自对甲苯磺酰基、邻硝基苯磺酰基或对硝基苯磺酰基；
所述烷基类氨基保护基选自三苯甲基、2,4-二甲氧基苄基、对甲氧基苄基或苄基；优选烷氧羰基类氨基保护基，更优选叔丁氧羰基或笏甲氧羰基。
所述的反应为缩合反应，形成酰胺键的缩合剂选自1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N,N'-二环己基碳化二亚胺、N,N'-二异丙基碳二酰亚胺、O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸酯、1-羟基苯并***、1-羟基-7-偶氮苯并三氮唑、O-苯并三氮唑-N,N,N',N'-四甲脲六氟磷酸酯、2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯、苯并三氮唑-1-基氧基三(二甲基氨基)磷鎓六氟磷酸盐或六氟磷酸苯并***-1-基-氧基三吡咯烷基磷，优选2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯；
上述缩合反应优选在碱性条件下进行，所述碱选自有机碱或无机碱，所述有机碱类优选三乙胺、二乙胺、N,N-二异丙基乙胺(二异丙基乙基胺)、吡啶、六甲基二硅基氨基钠，正丁基锂、叔丁醇钾或四丁基溴化铵，所述无机碱选自氢氧化锂、氢氧化钠、氢氧化钾、氢化钠、碳酸钠、碳酸氢钠、碳酸钾、碳酸氢钾或碳酸铯，更优选N,N-二异丙基乙胺。
优选的，所述的式(II)化合物，其中R 1为羧基保护基，R 2为氢原子；更优选的，所述的R 1羧基保护基为DMB(2,4-二甲氧基苄基)，所述的式(II)化合物如式(IIa)化合物所示。
优选的，所述的式(III)化合物，其中R 1为羧基保护基；更优选的，所述的R 1羧基保护基为DMB(2,4-二甲氧基苄基)，所述的式(III)化合物如式(IIIa)化合物所示；或者当所述的当R 1为氢原子时，所述的式(III)化合物如式(IIIb)化合物所示。
优选的，所述方法为
Figure PCTCN2018103781-appb-000006
其中式(IIa)所示化合物与6-马来酰亚氨基己酸发生缩合反应得到式(IIIa)所示化合物。
进一步优选的，所述方法还包括以下步骤
Figure PCTCN2018103781-appb-000007
式(IIIa)化合物经脱保护基反应，得到式(IIIb)化合物。
优选的，所述的式(II)化合物，当其中R 1为氢原子时，R 2为氨基保护基，所述的式(II)化合物如式(IIc)化合物所示；更优选的，所述的R 1羧基保护基为DMB(2,4-二甲氧基苄基)，R 2为氨基保护基，所述的式(II)化合物如式(IIb)化合物所示。
更优选的，所述方法还包括以下步骤
Figure PCTCN2018103781-appb-000008
上述方案中，所述方法还包括式(IIc)所示化合物加上羧基保护基后得到通式(IIb)所示化合物，式(IIb)所示化合物在碱性条件下脱Fmoc保护基得到式(IIa)所示化合物的步骤，
其中，
所述碱选自有机碱或无机碱，所述有机碱类优选三乙胺、二乙胺、N,N'-二异丙基乙胺、吡啶、六甲基二硅基氨基钠，正丁基锂、叔丁醇钾或四丁基溴化铵，所述无机碱选自氢氧化锂、氢氧化钠、氢氧化钾、氢化钠、碳酸钠、碳酸氢钠、碳酸钾、碳酸氢钾或碳酸铯，更优选二乙胺。
本发明还涉及式(I)所示化合物的制备方法，其特征在于，所述方法包括式(III)所示的化合物与式(IV)所示化合物反应得到式(V)所示化合物，式(V)所示化合物脱去保护基得到式(I)所示化合物的步骤，
Figure PCTCN2018103781-appb-000009
其中，
A选自3-8元环烷基，优选3元环烷基；
R选自卤素，优选氟、氯、溴或碘，最优选氟；
R 1如式(III)中定义；
R 3选自羧基保护基，所述羧基保护基选自2,4-二甲氧基苄基、苄基、烯丙基、五氟代苯基、甲基、对甲基苄基、甲氧乙氧甲基或叔丁基，优选2,4-二甲氧基苄基；
R x选自任选取代的烷基或环烷基，所述取代基选自烷基、卤代烷基、卤素、羟基、氨基、硝基、氰基、烷氧基、羟烷基、环烷基、杂环基、芳基或杂芳基；
n选自1、2、3、4或5。
优选的，所述方法还包括式(III)所示化合物在缩合剂的作用下与式(IV)所示化合物发生缩合反应得到式(V)所示化合物的步骤，
其中，
所述缩合剂选自1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N,N'-二环己基 碳化二亚胺、N,N'-二异丙基碳二酰亚胺、O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸酯、1-羟基苯并***、1-羟基-7-偶氮苯并三氮唑、O-苯并三氮唑-N,N,N',N'-四甲脲六氟磷酸酯、2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯、苯并三氮唑-1-基氧基三(二甲基氨基)磷鎓六氟磷酸盐或六氟磷酸苯并***-1-基-氧基三吡咯烷基磷，优选2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯。
上述缩合反应优选在碱性条件下进行，所述碱选自有机碱或无机碱，所述有机碱类优选三乙胺、二乙胺、N,N-二异丙基乙胺(二异丙基乙基胺)、吡啶、六甲基二硅基氨基钠，正丁基锂、叔丁醇钾或四丁基溴化铵，所述无机碱选自氢氧化锂、氢氧化钠、氢氧化钾、氢化钠、碳酸钠、碳酸氢钠、碳酸钾、碳酸氢钾或碳酸铯，更优选N,N-二异丙基乙胺。
更优选的，所述方法为
Figure PCTCN2018103781-appb-000010
优选的，所述方法包括式(IIIb)所示化合物与式(IVa)所示化合物在碱性条件下，缩合剂的作用下发生缩合反应得到式(Va)所示化合物，(Va)所示化合物在酸作用下脱去保护基得到式(Ia)所示化合物。
其中，缩合剂选自1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N,N'-二环己基 碳化二亚胺、N,N'-二异丙基碳二酰亚胺、O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸酯、1-羟基苯并***、1-羟基-7-偶氮苯并三氮唑、O-苯并三氮唑-N,N,N',N'-四甲脲六氟磷酸酯、2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯、苯并三氮唑-1-基氧基三(二甲基氨基)磷鎓六氟磷酸盐或六氟磷酸苯并***-1-基-氧基三吡咯烷基磷，优选2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯；
所述碱选自有机碱或无机碱，所述有机碱类优选三乙胺、二乙胺、N,N-二异丙基乙胺(二异丙基乙基胺)、吡啶、六甲基二硅基氨基钠，正丁基锂、叔丁醇钾或四丁基溴化铵，所述无机碱选自氢氧化锂、氢氧化钠、氢氧化钾、氢化钠、碳酸钠、碳酸氢钠、碳酸钾、碳酸氢钾或碳酸铯，更优选N,N-二异丙基乙胺。
提供酸性的条件的试剂包括但不限于氯化氢、氯化氢的1,4-二氧六环溶液、氯化铵、三氟乙酸、甲酸、乙酸、盐酸、硫酸、甲磺酸、硝酸、磷酸和对苯甲磺酸，优选三氟乙酸。
本发明还提供式(IVa)化合物的制备方法：
Figure PCTCN2018103781-appb-000011
式(M3)所示化合物在缩合剂的作用下与式(i)所示化合物发生酰胺化反应得到式(IVb)所示化合物，式(IVb)所示化合物在碱作用下脱去保护基得到式(IVa)所示化合物，
其中，
所述缩合剂选自1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N,N'-二环己基碳化二亚胺、N,N'-二异丙基碳二酰亚胺、O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸酯、1-羟基苯并***、1-羟基-7-偶氮苯并三氮唑、O-苯并三氮唑-N,N,N',N'-四甲脲六氟磷酸酯、2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯、苯并三氮唑-1-基氧基三(二甲基氨基)磷鎓六氟磷酸盐或六氟磷酸苯并***-1-基-氧基三吡咯烷基磷，优选2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯；
所述碱选自无机碱或有机碱，优选有机碱，有机碱选自胺类有机碱，例如二乙胺、三乙胺或氨水。
本发明还提供式(M3)化合物的制备方法：
Figure PCTCN2018103781-appb-000012
其中，式(M1)所示化合物在碱作用下与2,4-二甲氧基苄醇反应得到式(M2)所示化合物，式(M2)所示化合物在碱作用下脱去保护基得到式(M3)所示化合物；
所述碱选自无机碱或有机碱，优选有机碱，所述有机碱选自胺类有机碱，例如二乙胺、三乙胺、4-二甲氨基吡啶或1-(3-二甲基氨基丙基)-3-乙基碳二亚胺。
本发明进一步提供式(i)化合物的制备方法：
Figure PCTCN2018103781-appb-000013
其中，式(a)所示化合物在有机碱和有机硼作用下与式(b)所示化合物反应得到式(c)所示化合物，式(c)所示化合物在氧化剂(如双氧水)和碱(如一水氢氧化锂)作用下反应得到式(d)所示化合物，式(d)所示化合物在碱(如碳酸氢钠)作用与溴化苄反应得到式(e)所示化合物，式(e)所示化合物与碘甲烷反应得到式(f)所示化合物，式(f)所示化合物脱去保护基得到式(g)所示化合物，式(g)所示化合物与盐酸反应得到式(h)所示化合物，式(h)所示化合物在碱(如N,N-二异丙基乙胺)作用下与9-芴甲基-N-琥珀酰亚氨基羧酸酯反应得到式(i)所示化合物。
本发明还提供一种制备抗体药物偶联物或其药学上可接受盐的方法，其特征在于，所述抗体药物偶联物由抗体通过接头单元与上述方案中的通式(I)所示化合物偶联而成，所述抗体优选TNF-α抗体、VEGF抗体、c-MET抗体、EGFR抗体、Her2抗体或PD-1抗体，所述接头单元优选亚烃基、亚芳基、亚杂芳基、亚烃基硫烷基或亚氨基硫烷基。
优选的，所述TNF-α抗体选自依那西普、英夫利昔单抗、或阿达木单抗，VEGF抗体选自贝伐珠单抗，EGFR抗体选自西妥昔单抗、帕尼单抗或尼妥珠单抗，Her2抗体选自曲妥珠单抗(赫赛汀)或帕妥珠单抗，PD-1抗体选自纳武单抗或帕母单抗。
更优选的，所述抗体药物偶联物选自
Figure PCTCN2018103781-appb-000014
其中，抗体Ab优选TNF-α抗体、VEGF抗体、EGFR抗体、Her2抗体或PD-1抗体，y的范围为1-8，优选2-5；y是正实数，可以为整数或非整数。
发明详述
在本申请的说明书和权利要求书中，除非另有说明，否则本文中使用的科学和技术名词具有本领域技术人员所通常理解的含义。然而，为了更好地理解本发明，下面提供了部分相关术语的定义和解释。另外，当本申请所提供的术语的定义和解释与本领域技术人员所通常理解的含义不一致时，以本申请所提供的术语的定义和解释为准。
本发明所述“卤代”是指被“卤素原子”取代，“卤素原子”是指氟原子、氯原子、溴原子、碘原子等。
本发明所述“烷基”表示直链或支链的含有1-6个碳原子的烷基，包括例如“C 1-4烷基”、“C 1-3烷基”等，具体实例包括但不限于：甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、异戊基、2-甲基丁基、新戊基、1-乙基丙基、正己基、异己基、3-甲基戊基、2-甲基戊基、1-甲基戊基、3,3-二甲基丁基、2,2-二甲基丁基、1,1-二甲基丁基、1,2-二甲基丁基、1,3-二甲基丁基、2,3-二甲基丁基、2-乙基丁基、1,2-二甲基丙基等。
本发明所述的“卤代烷基”指一个或多个“卤素原子”取代“烷基”上的一个或多个氢原子所衍生的基团，所述“卤素原子”和“烷基”如前文所定义。
本发明所述的“羟基烷基”指一个或多个“羟基”取代“烷基”上的一个或多个氢原子所衍生的基团，所述“烷基”如前文所定义。
本发明所述的“烷氧基”是指以烷基-O-方式连接的基团，其中“烷基”如前文所定义。
本发明所述的“环烷基”指饱和或部分不饱和单环或多环环状基团，环原子个数包含3至20个，优选3至8个碳原子，更优选3至6个碳原子。具体实例包括但 不限于：环丙烷基、环丁烷基、环戊烷基、环己烷基、环庚烷基、环辛烷基等。
本发明所述的“杂环基”是指含有3-8个环原子(其中至少一个环原子为杂原子，例如氮原子、氧原子或硫原子)的环状基团。任选地，环状结构中的环原子(例如碳原子、氮原子或硫原子)可以被氧化，优选“5-6元杂环基”。具体实例包括但不仅限于：氮杂环丙烷基、2H-氮杂环丙烷基、二氮杂环丙烷基、3H-二氮杂环丙烯基、氮杂环丁烷基、1,4-二氧杂环己烷基、1,3-二氧杂环己烷基、1,3-二氧杂环戊烷基、1,4-二氧杂环己二烯基、四氢呋喃基、二氢吡咯基、吡咯烷基、咪唑烷基、4,5-二氢咪唑基、吡唑烷基、4,5-二氢吡唑基、2,5-二氢噻吩基、四氢噻吩基、4,5-二氢噻唑基、噻唑烷基、哌啶基、四氢吡啶基、哌啶酮基、四氢吡啶酮基、二氢哌啶酮基、哌嗪基、吗啉基、4,5-二氢噁唑基、4,5-二氢异噁唑基、2,3-二氢异噁唑基、噁唑烷基、2H-1,2-噁嗪基、6H-1,3-噁嗪基、4H-1,3-噻嗪基、6H-1,3-噻嗪基、2H-吡喃基、2H-吡喃-2-酮基、3,4-二氢-2H-吡喃基等。
本发明所述的“芳基”是指含有6-15个环碳原子的具有芳香性的环状基团，包括“6-8元芳基”、“6-15元稠芳基”，任选地，环状结构中的环原子(例如碳原子、氮原子或硫原子)可以被氧化。其实例包括但不限于：苯基、环辛四烯基、萘基、蒽基、菲基等。
本发明所述的“杂芳基”是指含有5-15个环原子(其中至少一个环原子为杂原子，例如氮原子、氧原子或硫原子)的具有芳香性的环状基团，包括“5-8元杂芳基”、“5-15元稠杂芳基”，任选地，环状结构中的环原子(例如碳原子、氮原子或硫原子)可以被氧化。具体实例包括但不限于：呋喃基、噻吩基、吡咯基、噻唑基、异噻唑基、噻二唑基、噁唑基、异噁唑基、噁二唑基、咪唑基、吡唑基、1,2,3-***基、1,2,4-***基、1,2,3-噁二唑基、1,2,4-噁二唑基、1,2,5-噁二唑基、1,3,4-噁二唑基、吡啶基、2-吡啶酮基、4-吡啶酮基、嘧啶基、哒嗪基、吡嗪基、1,2,3-三嗪基、1,3,5-三嗪基、1,2,4,5-四嗪基、氮杂环庚三烯基、1,3-二氮杂环庚三烯基、氮杂环辛四烯基、苯并呋喃基、苯并异呋喃基、苯并噻吩基、吲哚基、异吲哚基、苯并噁唑基、苯并咪唑基、吲唑基、苯并***基、喹啉基、2-喹啉酮基、4-喹啉酮基、1-异喹啉酮基、异喹啉基、吖啶基、菲啶基、苯并哒嗪基、酞嗪基、喹唑啉基、喹喔啉基、酚嗪基、喋啶基、嘌呤基、萘啶基、吩嗪基、吩噻嗪基等。
本发明所述的“接头单元”指包含使抗体共价附着于药物模块的共价键或原 子链的化学模块，例如，亚烃基、亚芳基、亚杂芳基、亚烃基硫烷基或亚氨基硫烷基，接头单元连同式(I)中的马来酰亚胺基团一起形成完整的接头。接头是本领域已知的，例如SMCC、SPP、SPDP等。
本发明所述“取代的”指基团中的一个或多个氢原子，优选为最多5个，更优选为1～3个氢原子彼此独立地被相应数目的取代基取代。不言而喻，取代基仅处在它们的可能的化学位置，本领域技术人员能够在不付出过多努力的情况下确定(通过实验或理论)可能或不可能的取代。例如，具有游离氢的氨基或羟基与具有不饱和(如烯属)键的碳原子结合时可能是不稳定的。
缩写词：
缩写 全称
Me 甲基
Boc 叔丁氧羰基
t-Bu 叔丁基
Bn 苄基
Ph 苯基
Tosyl 对甲基苯磺酰基
Fmoc 9-芴甲氧羰基
Alloc 烯丙氧羰基
Troc 三氯乙氧羰基
Cbz 苄氧羰基
PfP 五氟代苯基
PMB 对甲基苄基
MEM 甲氧乙氧甲基
Allyl 烯丙基
DMB 2,4-二甲氧基苄基
发明的有益效果
与现有技术(WO2005081711A2，公开日2005.09.09)相比，本发明制备式(I)所示化合物的技术方案具有以下优点：
(1)反应步骤缩短，本发明在制备式(I)所示化合物关键步骤中的反应物不同，本发明关键步骤中的第一步由反应物式(IIIb)所示化合物与式(IVa)所示化合物反应得到式(Va)所示化合物，第二步由式(Va)所示化合物脱去羧基保护基得到目标式(Ia)所示化合物，与现有技术比较，本发明的反应步骤缩短为两步，现有技术的反应步骤为四步；
(2)6-马来酰亚氨基己酰氯与片段连接的顺序不同；
(3)生产相同量的产物所需要反应物的投料量减少，本发明关键中间体式(IVa)所示化合物的投料量减少很多，本发明方法可以缩减生产成本，提高生产效率，具有突出的技术进步和显著的经济效果。
具体实施方式
以下将结合具体实例详细地解释本发明，使得本专业技术人员更全面地理解本发明，具体实例仅用于说明本发明的技术方案，并不以任何方式限定本发明。
实施例
化合物的结构是通过核磁共振(NMR)或/和质谱(MS)来确定的。NMR位移(δ)以10-6(ppm)的单位给出。NMR的测定是用Bruker AVANCE-400核磁仪，测定溶剂为氘代二甲基亚砜(DMSO-d6)、氘代氯仿(CDCl3)、氘代甲醇(CD3OD)，内标为四甲基硅烷(TMS)。
MS的测定用FINNIGAN LCQAd(ESI)质谱仪(生产商：Thermo，型号：Finnigan LCQ advantage MAX)。
HPLC的测定使用安捷伦1200DAD高压液相色谱仪(Sunfire C18 150×4.6mm色谱柱)和Waters 2695-2996高压液相色谱仪(Gimini C18 150×4.6mm色谱柱)。
手性HPLC分析测定使用LC-10A vp(Shimadzu)或者SFC-analytical(Berger Instruments Inc.)。
激酶平均抑制率及IC50值的测定用NovoStar酶标仪(德国BMG公司)。
薄层层析硅胶板使用烟台黄海HSGF254或青岛GF254硅胶板，薄层色谱法(TLC)使用的硅胶板采用的规格是0.15mm～0.2mm，薄层层析分离纯化产品采用的规格是0.4mm～0.5mm。
柱层析一般使用烟台黄海硅胶200～300目硅胶为载体。
手性制备柱层析使用Prep Star SD-1(Varian Instruments Inc.)或SFC-multigram(Berger Instruments Inc.)。
本发明的已知的起始原料可以采用或按照本领域已知的方法来合成，或可购买自ABCR GmbH&Co.KG、Acros Organics、Aldrich Chemical Company、韶远化学科技(Accela ChemBio Inc)、达瑞化学品等公司。
实施例中无特殊说明，反应能够均在氩气氛或氮气氛下进行。
氩气氛或氮气氛是指反应瓶连接一个约1L容积的氩气或氮气气球。
氢气氛是指反应瓶连接一个约1L容积的氢气气球。
加压氢化反应使用Parr 3916EKX型氢化仪和清蓝QL-500型氢气发生器或HC2-SS型氢化仪。
氢化反应通常抽真空，充入氢气，反复操作3次。
微波反应使用CEM Discover-S 908860型微波反应器。
实施例中无特殊说明，溶液是指水溶液。
实施例中无特殊说明，反应的温度为室温，为20℃～30℃。
实施例中的反应进程的监测采用薄层色谱法(TLC)，反应所使用的展开剂的体系有：A：二氯甲烷和甲醇体系，B：正己烷和乙酸乙酯体系，C：石油醚和乙酸乙酯体系，D：丙酮，溶剂的体积比根据化合物的极性不同而进行调节。纯化化合物采用的柱层析的洗脱剂的体系和薄层色谱法的展开剂体系包括：A：二氯甲烷和甲醇体系，B：正己烷和乙酸乙酯体系，C：二氯甲烷和丙酮体系，溶剂的体积比根据化合物的极性不同而进行调节，也可以加入少量的三乙胺和醋酸等碱性或酸性试剂进行调节。
实施例1
(S)-2-((2R,3R)-3-((1S,3S,5S)-2-((3R,4S,5S)-4-((S)-2-((S)-2-(6-(2,5-二氧代-2,5-二氢-1H-吡咯-1基)-N-甲基己酰氨基)-3-甲基丁酰氨基)-N,3-二甲基丁酰氨基)-3-甲氧基-5-甲基庚酰基)-2-氮杂二环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酰氨基)-3-(2-氟苯基)丙酸1
Figure PCTCN2018103781-appb-000015
Figure PCTCN2018103781-appb-000016
第一步
(1S,3S,5S)-叔丁基3-((1R,2R)-1-羟基-2-甲基-3-((4R,5S)-4-甲基-2-氧代-5-苯基噁唑烷-3-基)-3-氧代丙基)-2-氮杂双环[3.1.0]己烷-2-羧酸
将(4R,5S)-3-丙酰基-4-甲基-5-苯基-2-噁唑烷酮1b(14.7g，63.1mmol，采用专利申请“WO2015162291”公开的方法制备而得)溶解于二氯甲烷(120mL)中，冰浴冷 却至0℃，滴加三乙胺(9.7mL，72.6mmol)和三氟甲磺酸二丁硼(65mL，65mmol)，加毕，搅拌反应1小时。反应液于干冰-丙酮浴中冷却至-78℃，将(1S,3S,5S)-叔丁基3-甲酰基-2-氮杂双环[3.1.0]己烷-2-羧酸1a(12g，56.8mmol，采用专利申请“WO2008081399”公开的方法制备而得)溶解于二氯甲烷(10mL)中，向反应液中滴加上述溶液，加毕，搅拌反应2小时。撤去干冰丙酮浴，反应液自然升温至0℃，在0℃下，反应1.5小时。向反应液中加入磷酸盐缓冲液(pH＝7.0)和甲醇的混合溶液(100mL,V:V＝1:3)，再加入甲醇和双氧水(30％)的混合溶液(100mL,V:V＝2:1)，搅拌30分钟。反应液减压蒸除有机相，剩余水相用二氯甲烷(150mL×3)萃取，合并有机相，用水(100mL×2)洗涤，饱和氯化钠溶液(100mL×2)洗涤，用无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系C纯化所得残余物，得到标题产物1c(15.6g，产率72.3％)。
MS m/z(ESI):445.4[M+1]
第二步
(2R,3R)-3-((1S,3S,5S)-2-(叔丁氧羰基)-2-氮杂双环[3.1.0]己烷-3-基)-3-羟基-2-甲基丙酸
将1c(15.7g，35.3mmol)加入四氢呋喃(500mL)中，冰浴下，边缓慢滴加双氧水(15.3g，134mmol)边控制反应液温度在0℃，再加入一水氢氧化锂(2.5g，60mmol)，加毕，撤去冰浴，反应液温度自然升至室温，搅拌反应16小时。冰浴下，向反应液中分批缓慢加入亚硫酸钠(17.8g，141.2mmol)，加毕，搅拌反应30分钟，再加入水(150mL)，撤去冰浴，反应液减压蒸除有机相，剩余水相用二氯甲烷(150mL×3)洗涤，冰浴下，滴加2N的盐酸溶液至pH为2-3，用乙酸乙酯(100mL×3)萃取，用水(500mL)洗涤，饱和氯化钠溶液(500mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，得到标题产物1d(9.3g，产率92％)。
MS m/z(ESI):284.3[M-1]
第三步
(1S,3S,5S)-叔丁基3-((1R,2R)-3-(苄氧基)-1-羟基-2-甲基-3-氧代丙基)-2-氮杂双环[3.1.0]己烷-2-羧酸
将1d(9.3g，32.6mmol)溶解于N,N-二甲基甲酰胺(120mL)中，冰浴至0℃，加入碳酸氢钠(15.06g，179.3mmol)和溴化苄(27.9g，163.1mmol)，加毕，撤去冰浴，反应液于20℃搅拌16小时。冰浴下，向反应液中加入饱和氯化铵溶液(200mL) 淬灭反应，用乙酸乙酯(150mL×3)萃取反应液，有机相依次用水(200mL)和饱和氯化钠溶液(150mL)洗涤，用无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系C纯化所得残余物，得到标题产物1e(10.3g，产率84.2％)。MS m/z(ESI):376.4[M+1]
第四步
(1S,3S,5S)-叔丁基3-((1R,2R)-3-(苄氧基)-1-甲氧基-2-甲基-3-氧代丙基)-2-氮杂双环[3.1.0]己烷-2-羧酸
将1e(10.3g，27.4mmol)溶于N-甲基吡咯烷酮(90mL)中，加入碘甲烷(18mL，234mmol)，分批加入60％的氢化钠(1.28g，32.1mmol)，搅拌反应30分钟。冰浴下，将饱和氯化铵溶液(200mL)降温至0℃，加入反应液，搅拌10分钟，用乙酸乙酯(300mL×3)萃取，有机相依次用水(200mL×2)和饱和氯化钠溶液(200mL)洗涤，用无水硫酸钠干燥，过滤，滤液浓缩，用硅胶柱色谱法以洗脱剂体系C纯化所得残余物，得到标题产物1f(9.3g，产率87.2％)。
MS m/z(ESI):390.4[M+1]
第五步
(2R,3R)-3-((1S,3S,5S)-2-(叔丁氧羰基)-2-氮杂双环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酸
将1f(9.3g，23.88mmol)溶于四氢呋喃(90mL)中，加入钯碳(10％，1.86g)，氢气置换三次，反应液加热至35℃，搅拌反应30分钟。反应液经硅藻土过滤，滤液浓缩干燥，得到粗品标题产物1g(8.2g)，产品不经纯化直接进行下一步反应。
第六步
(2R,3R)-3-((1S,3S,5S)-2-氮杂双环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酸盐酸盐
将粗品1g(8.3g，27.7mmol)溶于氯化氢的1,4-二氧六环溶液(5.0M，50mL)中，搅拌反应30分钟。反应液减压浓缩干燥，得到粗品标题产物1h(8.2g)，产品不经纯化直接进行下一步反应。
第七步
(2R,3R)-3-((1S,3S,5S)-2-(((9H-芴-9-基)甲氧基)羰基)-2-氮杂双环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酸
将粗品1h(8.2g，41.2mmol)溶解于水(70mL)中，加入1,4-二氧六环(70mL)，冰浴下冷却至0℃，加入N,N-二异丙基乙胺(31.9g，247.2mmol)。将9-芴甲基-N- 琥珀酰亚氨基羧酸酯(18.06g，53.56mmol)溶解于1,4-二氧六环(70mL)中，将上述溶液加入到反应液中，加毕，撤去冰浴，反应液温度自然升至室温，搅拌反应2.5小时。反应液减压蒸除有机相，加入水(100mL)，冰浴下滴加1N的盐酸至反应液pH为2，用二氯甲烷(300mL×3)萃取，有机相依次用水(300mL×2)和饱和氯化钠溶液(300mL)洗涤，用无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系A纯化所得残余物，得到标题产物1i(8.7g，产率85％)。
MS m/z(ESI):422.4[M+1]
第八步
(S)-2,4-二甲氧基苄基2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-(2-氟苯基)丙酸
将(S)-2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-(2-氟苯基)丙酸1j(13g，32.6mmol)溶解于二氯甲烷(50mL)中，加入3,5-二甲氧基苯甲醇(5.93g，35.27mmol)，滴加N,N-二甲基甲酰胺二甲基缩醛(8.9mL，36.8mmol)，加毕，搅拌反应40小时。反应液依次用水(50mL)和饱和氯化钠溶液(50mL)洗涤，用无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系C纯化所得残余物，得到粗品标题产物1k(34.2g)，产品不经纯化直接进行下一步反应。
MS m/z(ESI):556.4[M+1]
第九步
(S)-2,4-二甲氧基苄基2-氨基-3-(2-氟苯基)丙酸
将1k(17g，30.6mmol)溶于二氯甲烷(40mL)中，加入二乙胺(80mL，0.78mmol)，搅拌反应3小时。反应液减压浓缩，用硅胶柱色谱法以洗脱剂体系A纯化所得残余物，得到标题产物1l(7.8g，产率76.4％)。
MS m/z(ESI):334.3[M+1]
第十步
(1S,3S,5S)-(9H-芴-9-基)甲基3-((1R,2R)-3-(((S)-1-((2,4-二甲氧基苄基)氧)-3-(2-氟苯基)-1-氧代丙基-2-基)氨基)-1-甲氧基-2-甲基-3-氧代丙基)-2-氮杂双环[3.1.0]己烷-2-羧酸
条件一：
将1i(2.5g，5.94mmol)和1l(2.04g，5.94mmol)溶于二氯甲烷和N,N-二甲基甲酰胺的混合溶液(36mL,V:V＝5:1)中，加入2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(2.7g，7.13mmol)和N,N-二异丙基乙胺(3.1mL，17.8mmol)，加毕，20℃搅拌反应1小时。向反应液中加入水(100mL)，搅拌，分层，有机相依次 用水(100mL×2)洗涤，饱和氯化钠溶液(100mL)洗涤，用无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系C纯化所得残余物，得到标题产物1m(3.9g，产率89.2％)。
条件二：
将化合物1i(9.1g，21.62mmol)，1l(7.5g，22.69mmol)和2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU，9.9g，25.94mmol)溶于二氯甲烷和N,N-二甲基甲酰胺的混合溶液(120mL，V:V＝5:1)中，加入N,N-二异丙基乙胺(8.4g，64.89mmol)，20℃反应1小时。向反应液中加入水(80mL)，搅拌分层，水相用二氯甲烷(100mL×2)萃取，合并有机相，用无水硫酸钠干燥，减压浓缩，用硅胶柱色谱法以洗脱剂体系C纯化所得残余物，得到标题产物1m(15.7g，产率98％)。
MS m/z(ESI):737.5[M+1]
第十一步
(S)-2,4-二甲氧基苄基2-((2R,3R)-3-((1S,3S,5S)-2-氮杂双环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酰胺)-3-(2-氟苯基)丙酸
条件一：
将1m(3.9g，5.29mmol)溶于二氯甲烷(20mL)中，加入二乙胺(40mL，0.39mmol)，20℃搅拌反应2小时。反应液减压蒸馏除去大部分二乙胺，加入甲苯(10mL)，减压浓缩干燥。所得残余物用二氯甲烷(100mL)溶解，依次用水(100mL×2)洗涤，饱和氯化钠溶液(100mL)洗涤，用无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系A纯化残留物，得到标题产物1n(2.31g，产率84.9％)。
条件二：
将1m(15.7g，21.3mmol)溶于二氯甲烷(100mL)中，于10-15℃缓慢加入二乙胺(150mL，1.5mmol)，搅拌反应3小时。反应液减压浓缩，用硅胶柱色谱法以洗脱剂体系A纯化所得残余物，得到标题产物1n(10g，产率91.7％)。
MS m/z(ESI):515.4[M+1]
第十二步
(5S,8S,11S,12R)-11-((S)-仲丁基)-1-(9H-芴-9-基)-5,8-二异丙基-12-甲氧基-4,10-二甲基-3,6,9-三氧代-2-氧杂-4,7,10-三氮杂十四-14-酸-2,4-二甲氧基苄酯1p
将(5S,8S,11S,12R)-11-((S)-仲丁基)-1-(9H-芴-9-基)-5,8-二异丙基-12-甲氧基 -4,10-二甲基-3,6,9-三氧代-2-氧杂-4,7,10-三氮杂十四-14-酸1o(5.13g，8.04mmol，采用专利申请“WO2013072813”公开的方法制备而得)溶于二氯甲烷(90mL)中，依次加入4-二甲氨基吡啶(100mg，0.8mmol)和1-(3-二甲基氨基丙基)-3-乙基碳二亚胺(2.31g，12.06mmol)，搅拌反应15分钟，滴加2,4-二甲氧基苄醇(1.62g，9.65mmol)的二氯甲烷溶液(10mL)，搅拌反应2小时。反应液中加入饱和氯化铵溶液(100mL)，搅拌，静置，分离有机相，有机相依次用饱和氯化铵溶液(100mL)、水(100mL)、饱和氯化钠溶液(100mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系C纯化所得残余物，得到标题产物1p(5.6g，产率88.5％)。MS m/z(ESI):788.8[M+1]
第十三步
(3R,5S)-4-((S)-N,3-二甲基-2-((S)-3-甲基-2-(甲氨基)丁酰氨基)丁酰氨基)-3-甲氧基-5-甲基庚酸-2,4-二甲氧基苄酯1q
将1p(4g，5.08mmol)溶于二氯甲烷(20mL)中，加入二乙胺(40mL)，搅拌反应3小时。反应液减压浓缩，用硅胶柱色谱法以洗脱剂体系A纯化所得残余物，得到标题产物1q(2.7g，产率94％)。
MS m/z(ESI):566.1[M+1]
第十四步
(3R,5S)-4-((S)-2-((S)-2-(6-(2,5-二氧代-2,5-二氢-1H-吡咯-1-基)-N-甲基己酰氨基)-3-甲基丁酰氨基)-N,3-二甲基丁酰氨基)-3-甲氧基-5-甲基庚酸-2,4-二甲氧基苄酯1r
将1q(2.7g，4.77mmol)溶于二氯甲烷和N,N-二甲基甲酰胺(48mL,V/V＝5：1)混合溶剂中，依次加入6-(2,5-二氧代-2,5-二氢-1H-吡咯-1-基)己酸(1.1g，5.25mmol，采用公知的方法“Journal of Medcinal Chemistry,2013,56(24),9955-9968”制备而得)、2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(2.18g，5.72mmol)和二异丙基乙基胺(3.3mL，19mmol)，搅拌反应1小时。反应液中加入水(100mL)，搅拌，静置，分离有机相，有机相依次用水(100mL×2)、饱和氯化钠溶液洗涤(100mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系C纯化所得残余物，得到标题产物1r(3g，产率82.9％)。
MS m/z(ESI):759.8[M+1]
第十五步
(3R,5S)-4-((S)-2-((S)-2-(6-(2,5-二氧代-2,5-二氢-1H-吡咯-1-基)-N-甲基己酰氨基)-3- 甲基丁酰氨基)-N,3-二甲基丁酰氨基)-3-甲氧基-5-甲基庚酸1s
(3R,5S)-4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-N-methylhexanamido)-3-methylbutanamido)-N,3-dimethylbutanamido)-3-methoxy-5-methylheptanoic acid
将1r(3g，3.95mmol)溶于三氟乙酸和二氯甲烷(50mL,V/V＝2.5％)混合溶剂中，搅拌反应15分钟。反应液中加入二氯甲烷，过滤，滤饼用二氯甲烷淋洗，滤液依次用水(100mL)、饱和氯化钠溶液洗涤(100mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，得到粗品标题产物1s(2.3g)，产物不经纯化直接进行下一步反应。MS m/z(ESI):609.7[M+1]
第十六步
(2S)-2-((2R,3R)-3-((1S,3S,5S)-2-((3R,5S)-4-((S)-2-((S)-2-(6-(2,5-二氧代-2,5-二氢-1H-吡咯-1-基)-N-甲基己酰氨基)-3-甲基丁酰氨基)-N,3-二甲基丁酰氨基)-3-甲氧基-5-甲基庚酰基)-2-氮杂二环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酰氨基)-3-(2-氟苯基)丙酸-2,4-二甲氧基苄酯1t
将粗品1s(2.3g，3.78mmol)和(S)-2-((2R,3R)-3-((1S,3S,5S)-2-氮杂二环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酰氨基)-3-(2-氟苯基)丙酸-2,4-二甲氧基苄酯1n(1.94g，3.78mmol)溶于二氯甲烷和N,N-二甲基甲酰胺(36mL,V/V＝5:1)混合溶剂中，依次加入2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(1.72g，4.54mmol)和二异丙基乙基胺(2.0mL，11.34mmol)，搅拌反应1小时。反应液中加入水(100mL)，搅拌，静置，分离有机相，有机相依次用水(100mL×2)、饱和氯化钠溶液洗涤(100mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系A纯化所得残余物，得到标题产物1t(1.94g，产率46.5％)。
MS m/z(ESI):1122.6[M+18]
第十七步
(2S)-2-((2R,3R)-3-((1S,3S,5S)-2-((3R,5S)-4-((S)-2-((S)-2-(6-(2,5-二氧代-2,5-二氢-1H-吡咯-1基)-N-甲基己酰氨基)-3-甲基丁酰氨基)-N,3-二甲基丁酰氨基)-3-甲氧基-5-甲基庚酰基)-2-氮杂二环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酰氨基)-3-(2-氟苯基)丙酸1
将1t(1.94g，1.75mmol)溶于三氟乙酸和二氯甲烷(40mL,V/V＝2.5％)混合溶剂中，搅拌反应15分钟。反应液减压浓缩，用硅胶柱色谱法以洗脱剂体系A纯化所得残余物，得到标题产物1(1.17g，产率69.8％)。
MS m/z(ESI):956.1[M+1]
1H-NMR(400MHz,CD 3OD):δ7.36-7.30(m,1H),7.29-7.21(m,1H),7.17-7.02(m,2H),6.83-6.79(m,2H),4.81-4.71(m,2H),4.69-4.55(m,2H),4.25-4.15(m,1H),4.13-4.04(m,1H),3.96-3.85(m,2H),3.70-3.61(m,1H),3.55-3.46(m,3H),3.40-3.21(m,4H),3.18-3.10(m,2H),3.07-2.96(m,4H),2.67-2.56(m,2H),2.54-2.34(m,3H),2.29-2.17(m,2H),2.10-1.99(m,1H),1.89-1.57(m,7H),1.52-1.28(m,6H),1.21-1.11(m,4H),1.07-0.96(m,6H),0.95-0.81(m,12H),0.80-0.69(m,1H)
对比例
采用现有技术的方法，直接用两个重要中间体连接生成式1所示化合物的方法：
Figure PCTCN2018103781-appb-000017
第1步
(2S)-2,4-二甲氧基苄基2-((2R,3R)-3-((1S,3S,5S)-2-((5S,8S,11S,12R)-11-((S)-仲丁基)-1-(9H-芴-9-基)-5,8-二异丙基-12-甲氧基-4,10-二甲基-3,6,9-三羰基-2-氧杂-4,7,10-三氮杂十四烷基-14-酰基)-2-氮杂双环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酰胺)-3-(2-氟苯基)丙酸酯
将1n(2.97g，5.77mmol)和(5S,8S,12R)-11-((S)-仲丁基)-1-(9H-芴-9-基)-5,8-二异丙基-12-甲氧基-4,10-二甲基-3,6,9-三羰基-2-氧杂-4,7,10-三氮杂十四烷-14-羧酸1o(3.68g，5.77mmol，采用专利申请“WO 2013072813”公开的方法制备而得)溶于 二氯甲烷和N,N-二甲基甲酰胺的混合溶液(48mL,V:V＝5:1)中，加入2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(2.63g，6.92mmol)和N,N-二异丙基乙胺(3mL，17.31mmol)，20℃搅拌反应1小时。加入水(100mL)，搅拌，分层，有机相依次用水(100mL×2)洗涤，饱和氯化钠溶液(100mL)洗涤，用无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系A纯化残留物，得到标题产物1p(5.3g，产率81.0％)。
MS m/z(ESI):1151.6[M+18]
第2步
(2S)-2,4-二甲氧基苄基2-((2R,3R)-3-((1S,3S,5S)-2-((3R,5S)-4-((S)-N,3-二甲基-2-((S)-3-甲基-2-(甲基氨基)丁酰氨基)丁酰氨基)-3-甲氧基-5-甲基庚酰基)-2-氮杂双环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酰氨基)-3-(2-氟苯基)丙酸酯
将1p(3.86g，3.4mmol)溶于二氯甲烷(20mL)中，加入二乙胺(40mL，0.39mmol)，20℃搅拌反应2小时。反应液减压蒸馏除大部分二乙胺，所得残余物中加入甲苯(10mL)，继续减压浓缩干燥。向所得残余物种加入水(100mL)和二氯甲烷(100mL)，搅拌，分液，有机相依次用水(100mL×2)洗涤，饱和氯化钠溶液(100mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系A纯化残留物，得到标题产物1q(2.73g，产率87％)。
MS m/z(ESI):912.9[M+1]
第3步
(2S)-2,4-二甲氧基苄基2-((2R,3R)-3-((1S,3S,5S)-2-((3R,5S)-4-((S)-2-((S)-2-(6-(2,5-二氧代-2,5-二氢-1H-吡咯-1-基)-N-甲基己酰氨基)-3-甲基丁酰氨基)-N,3-二甲基丁酰氨基)-3-甲氧基-5-甲基庚酰基)-2-氮杂双环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酰氨基)-3-(2-氟苯基)丙酸酯
将1q(1.7g，1.86mmol)溶于二氯甲烷和N,N-二甲基甲酰胺的混合溶液(30mL,V:V＝5:1)中，依次加入6-马来酰亚氨基己酸(0.39g，1.86mmol)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(0.85g，2.23mmol)和N,N-二异丙基乙胺(0.97mL，5.58mmol)，搅拌反应1小时。向反应液中加入50mL水(100mL)，搅拌，分层，有机相依次用水(50mL×2)洗涤，饱和氯化钠溶液(50mL)洗涤，用无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱色谱法以洗脱剂体系A纯化残留物，得到标题产物1r(1.01g，产率49％)。
MS m/z(ESI):1122.7[M+18]
第4步
(2S)-2-((2R,3R)-3-((1S,3S,5S)-2-((3R,5S)-4-((S)-2-((S)-2-(6-(2,5-二羰基-2,5-二氢-1H-吡咯-1-基)-N-甲基己酰氨基)-3-甲基丁酰氨基)-N,3-二甲基丁酰氨基)-3-甲氧基-5-甲基庚酰基)-2-氮杂双环[3.1.0]己烷-3-基)-3-甲氧基-2-甲基丙酰氨基)-3-(2-氟苯基)丙酸
将1r(2.5g，2.26mmol)溶于三氟乙酸的二氯甲烷溶液中(2.5％，50mL)，搅拌反应20分钟。冰浴下减压蒸除二氯甲烷，所得残余物减压干燥后用二氯甲烷溶解，湿法上样，用硅胶柱色谱法以洗脱剂体系A纯化，得到标题产物1(1.4g，产率64.8％)。
MS m/z(ESI):956.0[M+1]
1H-NMR(400MHz,CD 3OD):δ7.36-7.30(m,1H),7.29-7.21(m,1H),7.17-7.02(m,2H),6.83-6.79(m,2H),4.81-4.71(m,2H),4.69-4.55(m,2H),4.25-4.15(m,1H),4.13-4.04(m,1H),3.96-3.85(m,2H),3.70-3.61(m,1H),3.55-3.46(m,3H),3.40-3.21(m,4H),3.18-3.10(m,2H),3.07-2.96(m,4H),2.67-2.56(m,2H),2.54-2.34(m,3H),2.29-2.17(m,2H),2.10-1.99(m,1H),1.89-1.57(m,7H),1.52-1.28(m,6H),1.21-1.11(m,4H),1.07-0.96(m,6H),0.95-0.81(m,12H),0.80-0.69(m,1H).
表1、本发明实施例与现有技术对比例制备式1所示化合物的投料量比较
Figure PCTCN2018103781-appb-000018
结论：
由表1可知，本发明在第14步时6-马来酰亚氨基己酰氯与片段连接后，在生成相同量的目标式1所示化合物时，本发明与现有技术关键步骤使用的中间体不同，本发明采用中间体1s与1n反应(其中1s由1o制备得到)，对比例使用中间体1o与1n反应；本发明与现有技术比较，等量的中间体1o制备等量的式1化合物，所需要的中间体1n的量二者完全不相同，本发明中间体1n的量可以减少很多，中间体1n的量为1.94g，而现有技术为2.82g；本发明与现有技术关键反应步骤不相同，本发明为2步反应，现有技术为4步反应，现有技术需要经过多步复杂的 路线合成式1所示化合物，在采用本发明的合成方法后能大大降低生产成本，缩短反应时间、提高生产效率，具有突出的技术进步和显著的经济效果。

Claims (15)

  1. 一种通式(III)所示的化合物，
    Figure PCTCN2018103781-appb-100001
    其中：
    R 1选自氢原子或羧基保护基。
  2. 一种如权利要求1所述通式(III)所示化合物的制备方法，其特征在于，所述方法包括由式(II)所示化合物与6-马来酰亚氨基己酸反应得到式(III)所示化合物的步骤，
    Figure PCTCN2018103781-appb-100002
    其中，
    R 1如权利要求1中定义；
    R 2选自氢原子或氨基保护基。
  3. 如权利要求2所述的方法，其特征在于，所述方法为
    Figure PCTCN2018103781-appb-100003
  4. 如权利要求3所述的方法，其特征在于，所述方法还包括以下步骤
    Figure PCTCN2018103781-appb-100004
  5. 如权利要求3所述的方法，其特征在于，所述方法还包括以下步骤
    Figure PCTCN2018103781-appb-100005
  6. 如权利要求5所述的方法，其特征在于，式(IIb)所示化合物在碱性条件下脱Fmoc保护基得到式(IIa)所示化合物，其中，所述碱选自有机碱或无机碱，所述有机碱优选三乙胺、二乙胺、N,N'-二异丙基乙胺、吡啶、六甲基二硅基氨基钠，正丁基锂、叔丁醇钾或四丁基溴化铵，所述无机碱选自氢氧化锂、氢氧化钠、氢氧化钾、氢化钠、碳酸钠、碳酸氢钠、碳酸钾、碳酸氢钾或碳酸铯，更优选二乙胺。
  7. 一种通式(I)所示化合物的制备方法，其特征在于，所述方法包括式(III)所示的化合物与式(IV)所示化合物反应得到式(V)所示化合物，式(V)所示化合物脱去保护基得到式(I)所示化合物的步骤，
    Figure PCTCN2018103781-appb-100006
    其中，
    A选自3-8元环烷基，优选3元环烷基；
    R选自卤素，优选氟、氯、溴或碘，最优选氟；
    R 1如权利要求1中定义；
    R 3选自羧基保护基，所述羧基保护基优选2,4-二甲氧基苄基、苄基、烯丙基、五氟代苯基、甲基、对甲基苄基、甲氧乙氧甲基或叔丁基，优选2,4-二甲氧基苄基；
    R x选自任选取代的烷基或环烷基，所述取代基选自烷基、卤代烷基、卤素、羟基、氨基、硝基、氰基、烷氧基、羟基烷基、环烷基、杂环基、芳基或杂芳基；
    n选自1、2、3、4或5。
  8. 如权利要求7所述的方法，其特征在于，式(III)所示化合物在缩合剂的作用下与式(IV)所示化合物发生酰胺化反应得到式(V)所示化合物，所述缩合剂选自1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N,N'-二环己基碳化二亚胺、N,N'-二异丙基碳二酰亚胺、O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸酯、1-羟基苯并***、1-羟基-7-偶氮苯并三氮唑、O-苯并三氮唑-N,N,N',N'-四甲脲六氟磷酸酯、 2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯、苯并三氮唑-1-基氧基三(二甲基氨基)磷鎓六氟磷酸盐或六氟磷酸苯并***-1-基-氧基三吡咯烷基磷，优选2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯。
  9. 如权利要求7所述的方法，其特征在于，所述方法为
    Figure PCTCN2018103781-appb-100007
  10. 如权利要求9所述的方法，其特征在于，所述方法还包括以下步骤
    Figure PCTCN2018103781-appb-100008
  11. 如权利要求10所述的方法，其特征在于，所述方法还包括以下步骤
    Figure PCTCN2018103781-appb-100009
  12. 如权利要求10所述的方法，其特征在于，所述方法还包括以下步骤
    Figure PCTCN2018103781-appb-100010
  13. 一种制备抗体药物偶联物或其药学上可接受盐的方法，其特征在于，所述抗体药物偶联物由抗体通过接头单元与权利要求7所述的通式(I)所示化合物反应而成，所述抗体优选TNF-α抗体、VEGF抗体、EGFR抗体、c-MET抗体、Her2抗体或PD-1抗体。
  14. 如权利要求13所述的方法，其特征在于，所述TNF-α抗体选自依那西普、英夫利昔单抗、或阿达木单抗，VEGF抗体选自贝伐珠单抗，EGFR抗体选自西妥昔单抗、帕尼单抗或尼妥珠单抗，Her2抗体选自曲妥珠单抗(赫赛汀)或帕妥珠单抗，PD-1抗体选自纳武单抗或帕母单抗。
  15. 如权利要求13所述的方法，其特征在于，所述抗体药物偶联物选自
    Figure PCTCN2018103781-appb-100011
    其中，抗体Ab如权利要求13中所定义，y的范围为1-8，优选2-5。
PCT/CN2018/103781 2017-09-04 2018-09-03 一种新毒素及其中间体的制备方法 WO2019042447A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201880004457.6A CN109963835B (zh) 2017-09-04 2018-09-03 一种新毒素及其中间体的制备方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201710785396 2017-09-04
CN201710785396.9 2017-09-04

Publications (1)

Publication Number Publication Date
WO2019042447A1 true WO2019042447A1 (zh) 2019-03-07

Family

ID=65526239

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2018/103781 WO2019042447A1 (zh) 2017-09-04 2018-09-03 一种新毒素及其中间体的制备方法

Country Status (3)

Country Link
CN (1) CN109963835B (zh)
TW (1) TW201912626A (zh)
WO (1) WO2019042447A1 (zh)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109912684A (zh) * 2019-03-08 2019-06-21 联宁（苏州）生物制药有限公司 一种用于抗体药物偶联物的药物-连接子mc-mmaf的制备方法及其中间体
CN113813399A (zh) * 2021-04-19 2021-12-21 联宁（苏州）生物制药有限公司 喜树碱类化合物、其抗体药物偶联物及其应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104254342A (zh) * 2011-12-14 2014-12-31 西雅图基因公司 新抗体药物缀合物(adc)及其用途
WO2016127790A1 (zh) * 2015-02-15 2016-08-18 江苏恒瑞医药股份有限公司 配体-细胞毒性药物偶联物、其制备方法及其应用
WO2017144015A1 (zh) * 2016-02-26 2017-08-31 江苏恒瑞医药股份有限公司 一种新毒素及其中间体的制备方法
WO2018140275A2 (en) * 2017-01-26 2018-08-02 Seattle Genetics, Inc. Novel auristatin derivatives and related antibody-drug conjugates (adcs) and methods of preparation thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104254342A (zh) * 2011-12-14 2014-12-31 西雅图基因公司 新抗体药物缀合物(adc)及其用途
WO2016127790A1 (zh) * 2015-02-15 2016-08-18 江苏恒瑞医药股份有限公司 配体-细胞毒性药物偶联物、其制备方法及其应用
WO2017144015A1 (zh) * 2016-02-26 2017-08-31 江苏恒瑞医药股份有限公司 一种新毒素及其中间体的制备方法
WO2018140275A2 (en) * 2017-01-26 2018-08-02 Seattle Genetics, Inc. Novel auristatin derivatives and related antibody-drug conjugates (adcs) and methods of preparation thereof

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109912684A (zh) * 2019-03-08 2019-06-21 联宁（苏州）生物制药有限公司 一种用于抗体药物偶联物的药物-连接子mc-mmaf的制备方法及其中间体
WO2020181687A1 (zh) * 2019-03-08 2020-09-17 联宁（苏州）生物制药有限公司 一种用于抗体药物偶联物的药物-连接子mc-mmaf的制备方法及其中间体
KR20210125484A (ko) * 2019-03-08 2021-10-18 레베나 바이오파마 컴퍼니 리미티드 항체 약물 결합체에 사용되는 약물-링커 mc-mmaf의 제조 방법 및 이의 중간체
JP2022518601A (ja) * 2019-03-08 2022-03-15 聯寧（蘇州）生物製薬有限公司 抗体薬物複合体用薬物リンカーｍｃ－ｍｍａｆの調製方法及びその中間体
AU2019433421B2 (en) * 2019-03-08 2022-08-18 Levena Biopharma Co., Ltd. Preparation method for and intermediate of drug-linker for antibody drug conjugate MC-MMAF
JP7292751B2 (ja) 2019-03-08 2023-06-19 聯寧（蘇州）生物製薬有限公司 抗体薬物複合体用薬物リンカーｍｃ－ｍｍａｆの調製方法及びその中間体
KR102590042B1 (ko) * 2019-03-08 2023-10-17 레베나 (쑤저우) 바이오파마 컴퍼니 리미티드 항체 약물 결합체에 사용되는 약물-링커 mc-mmaf의 제조 방법 및 이의 중간체
CN113813399A (zh) * 2021-04-19 2021-12-21 联宁（苏州）生物制药有限公司 喜树碱类化合物、其抗体药物偶联物及其应用

Also Published As

Publication number Publication date
CN109963835B (zh) 2022-10-21
TW201912626A (zh) 2019-04-01
CN109963835A (zh) 2019-07-02

Similar Documents

Publication Publication Date Title
US20230145050A1 (en) Sulphonyl urea derivatives as nlrp3 inflammasome modulators
CA2958097C (en) Pyrrolopyrimidine compounds used as tlr7 agonist
US4923986A (en) Derivatives of physiologically active substance K-252
AU2018354972B2 (en) An improved process for the preparation of ribociclib and its salts
US8933225B2 (en) Process for the synthesis of E1 activating enzyme inhibitors
CA2971766A1 (en) Hemiasterlin derivatives for conjugation and therapy
JP2016513103A (ja) アルビシジン誘導体、その使用および合成
US10421716B2 (en) Process for preparing alpha-carboxamide pyrrolidine derivatives
JP4012145B2 (ja) ピロール−イミダゾールポリアミドの固相合成法
WO2019042447A1 (zh) 一种新毒素及其中间体的制备方法
WO2007037518A9 (ja) ムチリン誘導体及びそれを含有する医薬組成物
CN112979532B (zh) 邻苯二甲酰亚胺类化合物、制备方法和应用
TWI732826B (zh) 一種新毒素及其中間體的製備方法
France et al. Synthesis of a protected derivative of (2R, 3R)-β-hydroxyaspartic acid suitable for Fmoc-based solid phase synthesis
CN115190882A (zh) 通过共同中间体有效制备多拉司他汀和奥里斯他汀类似物
CN110028508B (zh) 一种抗肿瘤的重氮双环类细胞凋亡蛋白抑制剂
CA3095071A1 (en) Non-natural amatoxin-type antibody conjugate
EP3983387B1 (en) Sulfonylurea derivatives and uses thereof
US20230113609A1 (en) Sstr5 antagonists
CN111233661A (zh) 靶向泛素化降解ERRα蛋白的化合物及其药用组合物和应用
HARADA et al. New Route to 1, 3, 3a, 8a-Tetrahydro-2H-benzofuro [2, 3-b] pyrrol-2-ones from Methyl α-Hydroxy-4H-1, 2-benzoxazine-4-acetates Obtained by Ring Transformation of 4-Aryl-2-isoxazoline 2-Oxides
Songsichan Macrocyclization of omega-Isocyanoaldehydes and Towards the Total Synthesis of Jamaicensamide A
EP4208459A1 (en) Sulphamoyl urea derivatives containing alkyl-oxacycloalkyl moiety and uses thereof
JP2024511295A (ja) Ｃｙｐ１１ａ１阻害薬及びその中間体を調製する方法
TW202216662A (zh) 製備精胺酸酶抑制劑及其合成中間物之方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18849711

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18849711

Country of ref document: EP

Kind code of ref document: A1