Halogenated olefin camptothecin derivative, pharmaceutical composition, and preparation method therefor and use thereof

Abstract

The present invention relates to a camptothecin derivative, and a preparation method therefor and the use thereof in medicine. Specifically, the present invention relates to a camptothecin compound as shown in general formula (I), a preparation method therefor, a pharmaceutical composition containing the compound, and a conjugate thereof, and the use of the compound as a therapeutic agent, in particular as an inhibitor of topoisomerase I, and in the preparation of a drug for treating and / or preventing a disease or disorder by means of inhibiting topoisomerase I.

Description

Halogenated olefin camptothecin derivatives, pharmaceutical compositions, and preparation methods and applications thereof

[0001] This application claims priority to a prior application, patent application number 202410154407.3, filed with the State Intellectual Property Office of China on February 2, 2024, entitled “Halogenated olefin camptothecin derivatives, pharmaceutical compositions, preparation methods, and uses thereof.” The entirety of the prior application is incorporated herein by reference. Technical Field

[0002] The present invention belongs to the field of medicine, and in particular relates to a halogenated olefin camptothecin derivative, a pharmaceutical composition, and a preparation method and application thereof. Background Art

[0003] Camptothecin (CPT) is a natural product isolated from Camptotheca acuminata, a plant of the Davidiaceae family. It is a pentacyclic compound composed of a quinoline ring (AB), a pyrrole ring (C), a pyridone ring (D), and an α-hydroxylactone ring (E), with the 20-position in the S configuration (see the structural formula below). Due to its excellent anticancer activity, it was introduced to clinical practice in the early 1970s. However, clinical trials were terminated due to the development of severe side effects such as diarrhea and hemorrhagic cystitis.

[0004] Research data indicates that camptothecin can form a ternary complex with DNA topoisomerase I in cells, inhibiting DNA unwinding and leading to blocked DNA replication, which in turn causes cell death (Cancer Res. 1989, 49, 6365). Camptothecin and its derivatives have demonstrated potent antitumor activity in animal models of lung, breast, colorectal, and ovarian cancers (Nature Review Cancer. 2006, 6, 789). Currently, several camptothecin-based drugs have been approved for marketing for cancer treatment (Med Res. Rev. 2015, 35, 753). Irinotecan is used to treat colorectal cancer; topotecan is used to treat ovarian cancer; and belotecan is used to treat ovarian cancer and small cell lung cancer. Other camptothecin derivatives include ixitecan, rubitecan, diflutecan, lortotecan, gimatecan, ximintecan, gimitecan, and ilotecan. Camptothecin drugs or their derivatives often have hematotoxicity caused by bone marrow suppression, such as leukopenia, thrombocytopenia, anemia, neutropenia, etc., as well as gastrointestinal side effects such as nausea, vomiting, and diarrhea. Clinical studies have found that measures to improve the safety and efficacy of camptothecin compounds include increasing water solubility, improving their pharmacokinetic properties, increasing activity, reducing dosage, or using their conjugates with antibodies to form antibody-drug conjugates. Therefore, the development of camptothecin compounds and their conjugates with novel structures that can improve efficacy and ameliorate safety issues still has high clinical demand and application value. Summary of the Invention

[0005] The present invention provides a compound represented by Formula I, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound:

[0006] wherein Y is a halogen;

[0007] R1, R2, R3, R4 are the same or different and are independently selected from H, OH, CN, halogen, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 1-10 Alkoxy, halogenated C 1-10 Alkyl, halogenated C 1-10 Alkoxy, C 3-10 Cycloalkyl or halogenated C 3-10 Cycloalkyl;

[0008] R 51 、R 52 The same or different, independently selected from H, OH, CN, halogen, C 1-10 Alkyl, halogenated C 1-10 Alkyl, C 3-10 Cycloalkyl or halogenated C3-10 Cycloalkyl;

[0009] Alternatively, R2 and R3 together with the carbon atoms to which they are attached form C 3-10 Carbocyclic ring or 3-10 membered heterocyclic ring;

[0010] Alternatively, R3 and R4 together with the carbon atoms to which they are attached form C 3-10 Carbocyclic ring or 3-10 membered heterocyclic ring;

[0011] Alternatively, R4 and R 51 Together with the carbon atoms to which they are attached, they form C 5-10 Carbocyclic ring or 5-10 membered heterocyclic ring;

[0012] L1 is absent or selected from unsubstituted or optionally substituted by one, two or more selected from halogen, CN, C 1-10 Alkyl, halogenated C 1-10 Alkyl, or C 3-10 Substituents of cycloalkyl substituted C 1-10 alkylene;

[0013] X is selected from CR X1 R X2 NR X3 , O or S; R X1 、R X2 and R X3 the same or different, which are absent or independently selected from H or C 1-10 alkyl;

[0014] R6 is selected from H, halogen or C 1-10 alkyl;

[0015] R7 is selected from OH or NH2;

[0016] R 81 、R 82 The same or different, independently selected from H or C 1-10 alkyl;

[0017] Or, when X is selected from CR X1 R X2 or NR X3 When R X1 、R X2 or R X3 Any one and R 81 Together with their respective connected atoms, they form C 3-10 Carbocyclic ring or 3-10 membered heterocyclic ring;

[0018] R 91 、R 92 the same or different, independently selected from H, halogen, C 1-10 Alkyl, halogenated C1-10 Alkyl or C 3-10 Cycloalkyl;

[0019] Or, R 81 and R 91 Together with the carbon atoms to which they are attached, they form C 3-10 Carbocyclic ring or 3-10 membered heterocyclic ring;

[0020] wherein the carbocyclic or heterocyclic ring is unsubstituted or optionally substituted by one, two or more selected from halogen, CN, C 1-10 Alkyl, C 1-10 Alkoxy, halogenated C 1-10 Alkyl, halogenated C 1-10 Alkoxy, C 3-10 Cycloalkyl or halogenated C 3-10 Substitution of cycloalkyl groups;

[0021] m is selected from 0, 1, 2, 3, 4, 5 or 6.

[0022] According to some embodiments, Y is selected from F, Cl, Br.

[0023] According to some embodiments, Y is selected from F.

[0024] According to some embodiments, R1 is selected from H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 3-6 Cycloalkyl or halogenated C 3-6 Cycloalkyl.

[0025] According to some embodiments, R1 is selected from H.

[0026] According to some embodiments, R2 is selected from H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 3-6 Cycloalkyl or halogenated C 3-6 Cycloalkyl.

[0027] According to some embodiments, R2 is selected from H or F.

[0028] According to some embodiments, R3 is selected from H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 3-6 Cycloalkyl or halogenated C 3-6 Cycloalkyl.

[0029] According to some embodiments, R3 is selected from H or methyl.

[0030] According to some embodiments, R4 is selected from H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 3-6 Cycloalkyl or halogenated C 3-6 Cycloalkyl.

[0031] According to some embodiments, R4 is selected from H or Cl.

[0032] According to some embodiments, R 51 、R 52 The same or different, independently selected from H or C 1-4 alkyl.

[0033] According to some embodiments, R 51 、R 52 All are selected from H.

[0034] According to some embodiments, R2 and R3 together with the carbon atom to which they are attached form C 5-6 Carbocyclic ring or 5-6 membered heterocyclic ring.

[0035] According to some embodiments, R2 and R3 together with the carbon atoms to which they are attached form a 5-membered oxygen heterocycle (e.g. and in The position is fused with a benzene ring ).

[0036] According to some embodiments, R3 and R4 together with the carbon atom to which they are attached form a 5-membered carbocyclic ring (e.g. and in The position is fused with a benzene ring ).

[0037] According to some embodiments, R4 and R 51 Together with the carbon atoms to which they are attached, they form C 5-7 Carbocyclic ring or 5-7 membered heterocyclic ring.

[0038] According to some embodiments, R4 and R 51 Together with the carbon atoms to which they are attached, they form C 5-7 Carbocyclic ring or 5-7 membered oxygen-containing heterocyclic ring.

[0039] According to some embodiments, L1 is absent or selected from C 1-4 Alkylene.

[0040] According to some embodiments, L1 is absent or selected from methylene.

[0041] According to some embodiments, X is selected from CH2, NH or O.

[0042] According to some embodiments, R6 is selected from H or F.

[0043] According to some embodiments, R 81 、R 82 All are selected from H.

[0044] According to some embodiments, when X is selected from CR X1 R X2 or NR X3 When R X1 、R X2 or R X3 Any one and R 81 Together with their respective connected atoms, they form C 5-6 Carbocyclic ring or 5-6 membered N-containing heterocyclic ring.

[0045] According to some embodiments, when X is selected from NR X3 When R X3 and R 81 Together with the atoms to which they are attached, they form a tetrahydropyrrole ring or a piperidine ring.

[0046] According to some embodiments, R 91 、R 92 All are selected from H.

[0047] According to some embodiments, R 81 and R 91 Together with the carbon atoms to which they are attached, they form C 5-7 Carbocyclic ring or 5-7 membered heterocyclic ring.

[0048] According to some embodiments, R 81 and R 91 Together with the carbon atoms to which they are attached, they form a cyclopentane ring or a cyclohexane ring.

[0049] According to some embodiments, the carbocyclic or heterocyclic ring is unsubstituted or optionally substituted by one, two or more groups selected from halogen, CN, C 1-4 Alkyl (such as methyl, ethyl), C 1-4 Alkoxy (e.g. methoxy, ethoxy), halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 3-6 Cycloalkyl or halogenated C 3-6 The cycloalkyl group is substituted with a substituent.

[0050] According to some embodiments, m is selected from 0, 1, 2 or 3.

[0051] According to some embodiments, the compound structure shown in Formula I is as follows:

[0052] Among them, R1, R2, R3, R4, R 51 、R 52 , R7, R 81 、R 82 、R 91 、R 92 , L1, X, and m independently have the definitions described herein.

[0053] According to some embodiments, the compound structure shown in Formula I is as follows:

[0054] Among them, R1, R2, R3, R4, R6, R7, R 81 、R 82 、R 91 、R 92 , L1, X, m are independently defined as described herein;

[0055] X2 selected from CR X21 R X22 NR X23 , O or S; R X21 、R X22 and R X23 The same or different, independently selected from H or C 1-10 alkyl.

[0056] According to some embodiments, X2 is selected from CH2 or O.

[0057] According to some embodiments, the compound structure shown in Formula I is as follows:

[0058] wherein R1, R2, R3, R4, R6, R7, and X are independently defined as described herein.

[0059] According to an embodiment of the present invention, the structure of the compound of formula I is shown below:

[0060] According to some embodiments, the present invention also provides a structural fragment D of the compound of Formula I described herein after dehydrogenation.

[0061] According to some embodiments, the structure of D is as follows:

[0062] The present invention also provides a compound represented by Formula V, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound: L'-D (Formula V)

[0063] wherein L' is a linker containing a linker portion M capable of reacting with an antibody or an antigen-binding fragment thereof, and after L'-D reacts with the antibody or an antigen-binding fragment thereof, L' forms a linker L;

[0064] The definition of D is as shown in this article.

[0065] The present invention also provides an antibody drug conjugate of formula VI, Ab-[LD] β (Formula VI)

[0066] Wherein, Ab is an antibody or an antigen-binding fragment thereof, D has the definition as described in the text of the application, L is a linker connecting Ab and D, and β is selected from an integer or decimal between 1-10.

[0067] The present invention also provides a method for preparing the compound of formula I, comprising the following steps:

[0068] Among them, R1, R2, R3, R4, R 51 、R 52 、L1、X、Y、R6、R7、R 81 、R 82 、R 91 、R 92 and m are independently defined as described herein; Z is a leaving group (eg, F, Cl, Br).

[0069] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the compounds represented by Formula I or Formula V, their racemates, stereoisomers, tautomers, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds thereof.

[0070] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the antibody-drug conjugate represented by Formula VI.

[0071] According to an embodiment of the present invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

[0072] According to an embodiment of the present invention, the pharmaceutical composition may further contain one or more additional therapeutic agents.

[0073] The present invention also provides a method for treating tumor diseases, comprising administering to a patient a preventively or therapeutically effective amount of at least one of the compounds represented by Formula I or Formula V or the antibody-drug conjugate represented by Formula VI, their racemates, stereoisomers, tautomers, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds thereof.

[0074] The present invention also provides a method for treating tumor diseases, comprising administering to a patient a preventive or therapeutically effective amount of the above-mentioned pharmaceutical composition.

[0075] The tumor disease is selected from breast cancer, gastric cancer, lung cancer, colorectal cancer, large intestine cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal cancer, melanoma or leukemia.

[0076] In some embodiments, the patient comprises a mammal, preferably a human.

[0077] The present invention also provides at least one of the compounds represented by Formula I, Formula V, or the antibody-drug conjugate represented by Formula VI, their racemates, stereoisomers, tautomers, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds for treating tumor diseases, or pharmaceutical compositions thereof.

[0078] The present invention also provides the use of at least one of the compounds represented by Formula I, Formula V, or the antibody-drug conjugate represented by Formula VI, their racemates, stereoisomers, tautomers, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds thereof, or the pharmaceutical compositions thereof, in the preparation of topoisomerase I inhibitors and / or in the preparation of drugs for preventing or treating diseases or conditions associated with topoisomerase I.

[0079] In some embodiments, the disease or condition is a tumor, which includes breast cancer, gastric cancer, lung cancer, colorectal cancer, large intestine cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal cancer, melanoma or leukemia. Beneficial effects

[0080] The compounds or antibody-drug conjugates provided by the present invention have good tumor inhibitory effects and can be used to treat or prevent cancer, as well as to prepare drugs for treating or preventing such conditions and diseases.

[0081] Definitions and Explanations of Terms

[0082] Unless otherwise indicated, the definitions of groups and terms in this specification and claims, including definitions used as examples, exemplary definitions, preferred definitions, definitions in tables, and definitions of specific compounds in the Examples, may be arbitrarily combined and coupled with one another. The group definitions and compound structures resulting from such combinations and couplings should be understood to be within the scope of this specification and / or claims.

[0083] Unless otherwise indicated, numerical ranges recited in this specification and claims are equivalent to reciting at least each specific integer value therein. For example, the numerical range "1-12" is equivalent to reciting each integer value in the numerical range "1-12," namely, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12. In addition, when a numerical range is defined as a "number," it should be understood that both endpoints of the range, each integer within the range, and each decimal within the range are recited.

[0084] The term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0085] “C 1-10 "Alkyl" means straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, "C 1-8 "Alkyl" means straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms, "C 1-6 The term "alkyl" refers to straight-chain and branched alkyl groups having 1, 2, 3, 4, 5 or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, or the like or isomers thereof.

[0086] “C 2-10 "Alkenyl" is understood to mean preferably a linear or branched monovalent hydrocarbon radical containing one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, more preferably "C 2-8 Alkenyl". "C 2-10 "Alkenyl" is understood to mean preferably a linear or branched monovalent hydrocarbon radical containing one or more double bonds and having 2, 3, 4, 5, 6, 7 or 8 carbon atoms, e.g. 2-6 alkenyl), having 2 or 3 carbon atoms (i.e., C2-3 It is understood that when the alkenyl group contains more than one double bond, the double bonds may be separated from one another or conjugated. The alkenyl group is, for example, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, pent-4-enyl, (E)-pent-3-enyl, (Z)-pent-3-enyl, (E)-pent-2-enyl, (Z)-pent-2-enyl, (E)- Pent-1-enyl, (Z)-pent-1-enyl, hex-5-enyl, (E)-hex-4-enyl, (Z)-hex-4-enyl, (E)-hex-3-enyl, (Z)-hex-3-enyl, (E)-hex-2-enyl, (Z)-hex-2-enyl, (E)-hex-1-enyl, (Z)-hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl , 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl, (Z)-1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl, (E)-1-methylbut-2-enyl, (Z)-1-methyl But-2-enyl, (E)-3-methylbut-1-enyl, (Z)-3-methylbut-1-enyl, (E)-2-methylbut-1-enyl, (Z)-2-methylbut-1-enyl, (E)-1-methylbut-1-enyl, (Z)-1-methylbut-1-enyl, 1,1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.

[0087] The term "C 2-10 "Alkynyl" is understood to mean preferably a linear or branched monovalent hydrocarbon radical containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, e.g. having 2, 3, 4, 5, 6, 7 or 8 carbon atoms (i.e. "C 2-8 alkynyl”), having 2, 3, 4, 5, or 6 carbon atoms (i.e., “C 2-6 Alkynyl”), having 2 or 3 carbon atoms (“C 2-3The alkynyl group is, for example, ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hex-4-ynyl, hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-3-ynyl, 1-methylbut-2-ynyl, 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpent-4-ynyl, In some embodiments, the alkynyl group is ethynyl, prop-1-ynyl or prop-2-ynyl.

[0088] The term "C 3-10 "Cycloalkyl" is understood to mean a saturated monovalent monocyclic, bicyclic (eg bridged, spiro) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. 3-10 The cycloalkyl group may be a monocyclic hydrocarbon group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon group such as borneol, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonyl, 2,6-diazaspiro[3,4]octyl, or a tricyclic hydrocarbon group such as adamantyl.

[0089] Unless otherwise defined, the term "3-6 membered heterocyclyl" refers to a saturated or unsaturated non-aromatic ring or ring system, for example, a 4-, 5- or 6-membered monocyclic ring, and containing at least one, for example 1, 2, 3, 4, 5 or more heteroatoms selected from O, S and N, wherein N and S may also be optionally oxidized to various oxidation states to form nitrogen oxides, -S(O)- or -S(O)2- states. The heterocyclyl may include fused or bridged rings and spirocyclic rings. In particular, the heterocyclyl may include, but is not limited to, a 4-membered ring such as azetidinyl, oxetanyl; a 5-membered ring such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl.

[0090] The term "C 6-14 "Aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring ("C 6-14 or a ring having 9 carbon atoms ("C9 aryl"), for example indanyl or indenyl, or a ring having 10 carbon atoms ("C 10 aryl) such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms ("C 13 aryl), such as fluorenyl, or a ring having 14 carbon atoms ("C 14 aryl”), such as anthracenyl. When the C 6-20 When the aryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there is no limitation on the substitution position, and for example, the substitution may be at the ortho, para or meta position.

[0091] The term "5-14 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and which contain 1 to 5, preferably 1 to 3, heteroatoms each independently selected from N, O and S and which, in each case, may additionally be benzofused. "Heteroaryl" also refers to a radical in which a heteroaromatic ring is fused to one or more aryl, alicyclic or heterocyclyl rings, wherein the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7-, or 8-indolizinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4-, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinolizinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl, 2-, 3-, 4-, 5- or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl, 2-, 4-, 6- or 7-pteridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-4aHcarbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-carbazolylcarbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8- or 9-carbolinyl, 1-, 2-, 3- , 4-, 6-, 7-, 8-, 9- or 10-phenanthridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-acridinyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-piperidinyl, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenanthrolinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenothiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenanthrolinyl, , 3-, 4-, 5-, 6- or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-benzoisoquinolyl, 2-, 3-, 4- or thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-7H-pyrazino[2,3-c]carbazolyl, 2-, 3-, 5-, 6- or 7-2H-furo[3,2-b]pyranyl, 2-, 3-, 4-, 5-, 7- or 8-5H-pyrido[2,3-d]-o-oxazinyl, 1-, 3- or 5-1H-pyrazolo[4,3-d]-oxazolyl, 2-, 4- or 5-4H-imidazo[4,5-d]thiazolyl, 3-, 5- or 8-pyrazino[2,3-d]pyridazinyl, 2-, 3-, 5- or 6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8- or 9-furo[3,4-c]cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10 or 11-4H-pyrido[2,3-c]carbazolyl, 2-, 3-, 6- or 7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thiazolyl phenyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzoimidazolyl, 2-, 4-, 4-, 5-, 6-, or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7-, or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 11-4H-pyrrolo[1,2-b][2]benzazapinyl. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl. When the 5-20 membered heteroaryl group is linked to other groups to form the compounds of the present invention, the carbon atoms on the 5-20 membered heteroaryl ring may be linked to the other groups, or heteroatoms on the 5-20 membered heteroaryl ring may be linked to the other groups. When the 5-20 membered heteroaryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there is no limitation on the substitution site, for example, a hydrogen atom connected to a carbon atom on a heteroaryl ring may be substituted, or a hydrogen atom connected to a heteroatom on a heteroaryl ring may be substituted.

[0092] The term "spirocyclic" refers to a ring system in which two rings share one ring atom.

[0093] The term "fused ring" refers to a ring system in which two rings share two ring atoms.

[0094] The term "bridged ring" refers to a ring system in which two rings share three or more ring atoms.

[0095] Unless otherwise specified, a heterocyclic group, heteroaryl group, or heteroarylene group includes all possible isomeric forms thereof, such as positional isomers thereof. Thus, for some illustrative non-limiting examples, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-positions, etc. (if present) may include 1, 2, or more substituted or bonded forms thereof, including pyridin-2-yl, pyridin-2-ylene, pyridin-3-yl, pyridin-3-ylene, pyridin-4-ylene, and pyridin-4-ylene; thienyl or thienylene group includes thien-2-yl, thien-2-ylene, thien-3-ylene, and thien-3-ylene; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.

[0096] The term "alkyloxy" refers to -O-(alkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy include methoxy, ethoxy, propoxy, butoxy. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkyloxy, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, or heterocycloalkyloxy.

[0097] The term "alkylamino" refers to -NH-(alkyl), wherein alkyl is as defined above. Non-limiting examples of alkylamino include methylamino, ethylamino, propylamino, isopropylamino, butylamino, and the like.

[0098] The term "(alkyl)2amino" refers to -N-(alkyl)2, wherein alkyl is as defined above. Non-limiting examples of (alkyl)2amino include dimethylamino, methylethylamino, diethylamino, dipropylamino, methylpropylamino, diisopropylamino, dibutylamino, and the like.

[0099] "Haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

[0100] In the general formula or compound structure of the present application, an atom connected to a carbon atom in a carbon-carbon double bond (C=C) by a wavy line indicates that the position of the atom on one side of the carbon-carbon double bond is not fixed, that is, it can form either an E configuration or a Z configuration. It can represent structure It can also represent structure Unless otherwise stated, similar expressions in this application shall be interpreted the same as above.

[0101] Wavy lines intersecting chemical bonds Used to indicate the connection position between a group and other atoms in the molecular structure. Indicates connection with the 3-position of pyridyl. When the group connection position is not fixed, taking pyridyl as an example, The above-mentioned embodiment shows that the pyridyl group can be connected to any position on the pyridyl group. Unless otherwise specified, similar expressions in this application are interpreted the same as above.

[0102] It will be appreciated by those skilled in the art that the compounds of formula (I) may exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they may form acid addition salts; if these compounds have an acidic center, they may form base addition salts; if these compounds contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amino group), they may also form internal salts.

[0103] The compounds of the present invention may exist in the form of solvates (e.g., hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0104] Depending on their molecular structure, the compounds of the present invention may be chiral and therefore may exist in various enantiomeric forms. Thus, these compounds may exist in racemic or optically active forms. The compounds of the present invention encompass isomers or mixtures thereof, racemates, in which each chiral carbon is in the R or S configuration. The compounds of the present invention or their intermediates can be separated into enantiomeric compounds by chemical or physical methods well known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are prepared from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, appropriate N-protected amino acids (e.g., N-benzoylproline or N-phenylsulfonylproline) or various optically active camphorsulfonic acids. Chromatographic enantiomer resolution can also be advantageously performed with the aid of optically active resolving agents (e.g., dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chirally derivatized methacrylate polymers immobilized on silica gel). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, e.g., hexane / isopropanol / acetonitrile.

[0105] The corresponding stable isomers can be separated according to known methods, for example by extraction, filtration or column chromatography.

[0106] The term "patient" refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans.

[0107] The term "therapeutically effective amount" refers to that amount of an active compound or drug that will elicit the biological or medical response that a researcher, veterinarian, physician, or other clinician is seeking in a tissue, system, animal, individual, or human, and includes one or more of the following: (1) prevents disease, e.g., prevents a disease, disorder, or condition in an individual who is susceptible to the disease, disorder, or condition but who is not yet experiencing or developing the pathology or symptoms of the disease. (2) inhibits disease, e.g., inhibits the disease, disorder, or condition (i.e., prevents further development of the pathology and / or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder, or condition. (3) alleviates disease, e.g., alleviates the disease, disorder, or condition (i.e., reverses the pathology and / or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder, or condition. DETAILED DESCRIPTION

[0108] The technical solutions of the present invention will be described in further detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are encompassed within the scope of protection that the present invention is intended to protect.

[0109] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0110] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of ppm is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), and tetramethylsilane (TMS) was used as the internal standard.

[0111] MS measurements were performed using an Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS liquid chromatography-mass spectrometer (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS), a Waters ACQuity UPLC-QD / SQD (manufacturer: Waters, MS model: Waters ACQuity Qda Detector / Waters SQ Detector), and a THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model: THERMO Q Exactive).

[0112] High performance liquid chromatography (HPLC) analysis was performed using Agilent 1260II HPLC and Waters Acquity UPLC H-Class HPLC.

[0113] Chiral HPLC analysis was performed using a Waters Acquity UPCC high performance liquid chromatograph.

[0114] High-performance liquid chromatography (HPLC) was performed using Waters MS-triggered Prep-LC with SQD2 detector, Waters MS triggered Prep-LC with Acquity QDA detector, Waters MS-triggered Prep-LC with QDA detector, and GILSON Prep LC with UV detector.

[0115] The CombiFlash rapid preparation instrument used was Combiflash Rf200 (TELEDYNE ISCO).

[0116] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm~0.2mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm~0.5mm.

[0117] Silica gel column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0118] Average kinase inhibition rate and IC 50 The values ​​were determined using a NovoStar microplate reader (BMG, Germany).

[0119] The starting materials known in the present invention can be synthesized by methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Darui Chemicals, and other companies.

[0120] Unless otherwise specified in the examples, all reactions can be carried out under an argon atmosphere or a nitrogen atmosphere.

[0121] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1 L.

[0122] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

[0123] The pressurized hydrogenation reaction uses a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0124] The hydrogenation reaction is usually carried out by evacuating the chamber and filling it with hydrogen, and the operation is repeated three times.

[0125] A CEM Discover-S 908860 microwave reactor was used for the microwave reaction.

[0126] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0127] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0128] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the column chromatography eluent system used for the purified compounds, and the developing solvent system for thin layer chromatography included: A: dichloromethane / methanol system, B: n-hexane / ethyl acetate system. The volume ratio of the solvents was adjusted according to the polarity of the compounds, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid were optionally added for adjustment.

[0129] Example 1

[0130] (S)-11-(((3,3-difluoro-2-(hydroxymethyl)allyl)amino)methyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione

[0131] The first step: N-(3-fluoro-4-methylphenyl)acetamide (001b)

[0132] Under an ice bath, 3-fluoro-4-methylaniline 001a (25 g, 0.2 mol) was added portionwise to a mixture of acetic anhydride (75 mL) and concentrated sulfuric acid (0.5 mL). The reaction was then stirred at 10°C for 30 minutes. After completion of the reaction, the mixture was poured into ice water to precipitate a solid, which was filtered and washed three times with water. The filter cake was collected and dried to yield compound 001b (30 g), which was used directly in the next reaction without further purification.

[0133] MS m / z(ESI):168.1(M+1) + .

[0134] Step 2: N-(2-bromo-5-fluoro-4-methylphenyl)acetamide (001c)

[0135] Compound 001b (17 g, 0.10 mol) and sodium acetate (10 g, 0.12 mol) were added to acetic acid (40 mL) and heated to 60°C. A solution of bromine (19.5 g, 0.12 mol) in acetic acid (40 mL) was then added. The reaction was stirred at 80°C for 2 hours. After completion of the reaction, the mixture was poured into ice water to precipitate a solid, which was filtered and washed three times with water. The filter cake was collected and dried to obtain the crude product, Compound 001c (23 g), which was used directly in the next reaction without further purification.

[0136] MS m / z(ESI):246.0(M+1) + .

[0137] Step 3: N-(2-(1-ethoxyvinyl)-5-fluoro-4-methylphenyl)acetamide (001d)

[0138] Compound 001c (4 g, 16.26 mmol) was dissolved in 1,4-dioxane (40 mL), and tributyl-(1-ethoxyvinyl)tin (7.06 g, 19.56 mmol) and bis(triphenylphosphine)palladium dichloride (230 mg, 0.33 mmol) were added. Under nitrogen, the reaction was stirred at 100°C for 2 hours. After completion of the reaction, the filtrate was filtered, collected, and the solvent was evaporated to dryness to obtain a crude product. The crude product was purified by column chromatography using System B to obtain Compound 001d (2.6 g, 67% yield).

[0139] MS m / z(ESI):238.0(M+1) + .

[0140] Step 4: N-(2-acetyl-5-fluoro-4-methylphenyl)acetamide (001e)

[0141] Compound 001d (2.5 g, 10.5 mmol) was dissolved in a solution of hydrochloric acid in dioxane (20 mL, 4 mol / L). The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the mixture was neutralized with saturated sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and the solvent was evaporated to give compound 001e (1.9 g, 87% yield).

[0142] MS m / z(ESI):210.0(M+1) + .

[0143] Step 5: N-(2-(2-bromoacetyl)-5-fluoro-4-methylphenyl)acetamide (001f)

[0144] Compound 001e (1.5 g, 7.2 mmol) was dissolved in acetic acid (20 mL), and a solution of hydrobromic acid in acetic acid (14.4 mmol, 33% wt) was slowly added. Liquid bromine (1.15 g, 7.2 mmol) was then slowly added. The reaction was stirred at room temperature for half an hour. After completion of the reaction, the mixture was poured into ice water to precipitate a solid, which was filtered and washed three times with water. The filter cake was collected and dried to obtain the crude product, Compound 001f (1.7 g, 82% yield). The product was used directly in the next reaction without further purification.

[0145] MS m / z(ESI):288.0(M+1)+.

[0146] Step 6 1-(2-amino-4-fluoro-5-methylphenyl)-2-bromoethane-1-one (0.01 g)

[0147] Compound 001f (1.5 g, 5.2 mmol) was dissolved in ethanol (20 mL) and hydrobromic acid (2 mL) was added. The reaction was stirred at 80°C for 1 hour. After completion of the reaction, the system was adjusted to neutrality with saturated sodium bicarbonate aqueous solution, then extracted with ethyl acetate. The organic phase was collected and the solvent was evaporated to obtain the crude product, Compound 001g (1.1 g), which was used directly in the next reaction without purification.

[0148] MS m / z(ESI):246.0(M+1) + .

[0149] Step 7: (S)-11-(bromomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizine[1,2-b]quinoline-3,14(4H)-dione (001h)

[0150] Compound 001g (500 mg, 2.03 mmol) was dissolved in toluene (10 mL), and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (588 mg, 2.24 mmol) and p-toluenesulfonic acid monohydrate (463 mg, 2.44 mmol) were added. The reaction was stirred at 100°C for 1 hour. After completion of the reaction, the solvent was evaporated to obtain a crude product, which was purified by column chromatography using System B to obtain Compound 001h (500 mg, 52% yield).

[0151] MS m / z(ESI):473.0(M+1) + .

[0152] Step 8: Tert-butyl (2-(((tert-butyldiphenylsilyl)oxy)methyl)-3,3-difluoroallyl)carbamate (001j)

[0153] Tert-butyl (3-((tert-butyldiphenylsilyl)oxy)-2-oxopropyl)carbamate 001i (300 mg, 0.70 mmol) was dissolved in N,N-dimethylformamide (2 mL), and difluoromethyl (2-pyridyl) sulfone (163 mg, 0.84 mmol) was added. At -50 ° C, a solution of potassium tert-butoxide in tetrahydrofuran (0.1 mL, 1 mol / L) was slowly added dropwise. The temperature was then raised to room temperature and stirred for 1 hour. After the reaction was completed, water was added, and the system was extracted with dichloromethane. The organic phase was collected and the solvent was dried to obtain a crude product. The crude product was purified by column chromatography separation system B to obtain compound 001j (70 mg, yield: 25%).

[0154] MS m / z(ESI):484.2(M+1) + .

[0155] Step 9: 2-((tert-Butyldiphenylsilyl)oxy)methyl)-3,3-difluoroprop-2-en-1-amine (001k)

[0156] Compound 001j (70 mg, 0.15 mmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (0.2 mL) was added. The reaction was stirred at room temperature for 1 hour. After the reaction, the solvent was evaporated to obtain a crude product, compound 001k (53 mg), which was used directly in the next reaction without purification.

[0157] MS m / z(ESI):362.0(M+1) + .

[0158] Step 10: (S)-11-((2-(((tert-butyldiphenylsilyl)oxy)methyl)-3,3-difluoroallyl)amino)methyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizine[1,2-b]quinoline-3,14(4H)-dione (0011)

[0159] Compound 001k (53 mg, 0.16 mmol) and compound 001h (50 mg, 0.11 mmol) were dissolved in N,N-dimethylformamide (5 mL), and N,N-diisopropylethylamine (41 mg, 0.32 mmol) was added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the mixture was spin-dried to obtain compound 001l (79 mg), which was used directly in the next step without purification.

[0160] MS m / z(ESI):754.2(M+1) + .

[0161] Step 11 (S)-11-(((3,3-difluoro-2-(hydroxymethyl)allyl)amino)methyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizine[1,2-b]quinoline-3,14(4H)-dione (001)

[0162] Compound 0011 (79 mg, 0.1 mmol) was dissolved in 4N hydrochloric acid in methanol (2 mL). The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was purified using preparative HPLC (Waters MS-triggered Prep-LC equipped with a QDA detector, column: Gemini 5μm C18 100 x 21.2 mm, 5μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 10-minute gradient, gradient ratio: acetonitrile phase 20%-30%, flow rate: 25 mL / min) to obtain compound 001 (9.6 mg, two-step yield: 17%).

[0163] MS m / z(ESI):516.2(M+1) + .

[0164] 1 H NMR(400MHz,DMSO-d6)δ9.44(s,1H),8.45(d,1H),8.00(d,1H),7.36(s,1H),6.56(s,1H),5.58(s,2H),5.46(s ,2H),4.87(s,1H),4.19(s,2H),4.03(s,1H),2.56(s,3H),2.04–1.95(m,1H),1.92–1.81(m,2H),0.88(t,3H).

[0165] Example 2

[0166] (S)-4-Ethyl-8-fluoro-11-((3-fluoro-2-(hydroxymethyl)allyl)amino)methyl)-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione

[0167] Step 1: Tert-butyl (2-(((tert-butyldiphenylsilyl)oxy)methyl)-3-fluoroallyl)carbamate (002a)

[0168] (Fluoromethyl)triphenylphosphine tetrafluoroborate (382 mg, 1.0 mmol) and sodium bis(trimethylsilyl)amide (183 mg, 1.0 mmol) were dissolved in tetrahydrofuran (10 mL) and stirred at -78°C for 1 hour. Compound 001i (300 mg, 0.7 mmol) was slowly added dropwise, and the reaction mixture was allowed to warm to room temperature and stirred overnight. LCMS confirmed the reaction was complete. The reaction mixture was poured into water and extracted three times with ethyl acetate, washed once with saturated brine, and concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography on System B to afford Compound 002a (150 mg, 48% yield).

[0169] MS m / z(ESI):466.2(M+Na) + .

[0170] Step 2: 2-(((tert-Butyldiphenylsilyl)oxy)methyl)-3-fluoroprop-2-en-1-amine (002b)

[0171] Compound 002a (150 mg, 0.32 mmol) was dissolved in dioxane hydrochloride solution (10 mL), stirred at room temperature for 2 hours, and the solvent was evaporated under reduced pressure to obtain a crude product, compound 002b (105 mg), which was directly used for the next reaction without purification.

[0172] MS m / z(ESI):344.2(M+1) + .

[0173] Step 3: (S)-11-((2-(((tert-butyldiphenylsilyl)oxy)methyl)-3-fluoroallyl)amino)methyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizine[1,2-b]quinoline-3,14(4H)-dione (002c)

[0174] Compound 002b (50 mg, 0.15 mmol) was dissolved in N,N-dimethylformamide (10 mL), and compound 001h (52 mg, 0.11 mmol) and N,N-diisopropylethylamine (23.4 mg, 0.18 mmol) were added. The reaction mixture was heated to 50°C and stirred for 6 hours. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain the crude product, compound 002c (75 mg), which was used directly in the next step without purification.

[0175] MS m / z(ESI):736.3(M+1) + .

[0176] Step 4: (S)-4-ethyl-8-fluoro-11-((3-fluoro-2-(hydroxymethyl)allyl)amino)methyl)-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (002)

[0177] Compound 002c (75 mg, 0.1 mmol) was dissolved in 4 M hydrochloric acid and dioxane solution (2 mL). The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was purified using preparative HPLC (Waters MS-triggered Prep-LC equipped with an SQD2 detector, column: Xbridge 5u C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 13-minute gradient, gradient ratio: acetonitrile phase 18% to 28%, flow rate: 20 mL / min) to obtain compound 002 (2 mg, yield: 4%).

[0178] MS m / z(ESI):498.2(M+1) + .

[0179] 1 H NMR(400MHz,DMSO-d6)δ8.28(d,1H),7.86(d,1H),7.29(s,1H),6.89(d,1H),6.50(s,1H),5.41(s,2H),5.3 7(s,2H),4.84(s,1H),4.22(s,2H),3.95(s,2H),3.36(d,2H),2.50(s,3H),1.89-1.79(m,2H),0.85(t,3H).

[0180] Example 3

[0181] (S)-7-Ethyl-14-((3-fluoro-2-(hydroxymethyl)allyl)amino)methyl)-7-hydroxy-10,13-dihydro-11H-[1,3]dioxetine[4,5-g]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-8,11(7H)-dione

[0182] Compound 003e was prepared using compound 003a as the starting material using a method similar to that used in the synthesis of 001h in Example 1.

[0183] Compound 003e and compound 002b were used as starting materials, using a synthesis method similar to that in Example 2, and purified by HPLC (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5u C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 15%-25%, flow rate: 20 mL / min) to obtain compound 003 (2 mg, yield: 5%).

[0184] MS m / z(ESI):510.1(M+1) + .

[0185] 1 H NMR (400MHz, CD3OD) δ8.21-8.03(m,1H),7.72(s,1H),7.55(d,1H),6.26(s,1H),6.05(s,2H),5. 38(s,2H),5.33(t,2H),4.21(s,2H),3.33(d,2H),2.21-2.16(m,2H),1.60(s,2H),0.88(t,3H).

[0186] Biological evaluation

[0187] The present invention is further described and explained below in conjunction with test examples, but these test examples are not intended to limit the scope of the present invention.

[0188] The structure of the comparative compound (Dxd, commercially available) used in the test is as follows:

[0189] Test Example 1 Inhibitory experiment of compound on SK-BR-3 cell proliferation

[0190] 1.1 Experimental Materials

[0191] 1.2 Experimental instruments

[0192] 1.3 Test Method

[0193] (1) Cell plating: First, culture the tumor cells SK-BR-3 in the corresponding culture medium, digest the cells with trypsin, resuspend the cells after centrifugation, count the cells, adjust the cells to the appropriate concentration, and plate them on a 384-well plate.

[0194] (2) Co-incubation of compounds and tumor cells: After cells adhered, 100 nL of diluted bioactive substance (test compound) was added to the cell culture plate using ECHO. The final DMSO concentration in the cell plate wells was 0.33%. The cells were incubated in a 37°C, 5% CO2 incubator for 72 h.

[0195] (3) After the incubation, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the wells were shaken on a rapid shaker for 2 minutes, centrifuged at 1000 rpm for 1 minute, and placed in the dark at room temperature for 30 minutes. The chemiluminescence signal value was read using an Envision instrument.

[0196] (4) Cell viability assay: IC was calculated using GraphPad Prism 8 software 50 , the IC of the compound was obtained using the following nonlinear fitting formula: 50 (See Table 1):

[0197] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0198] Y: inhibition rate; X: log value of compound concentration;

[0199] Inhibition rate (%) = 100 - (reading value of compound well - reading value of low reading control well) / (reading value of high reading control well - reading value of low reading control well) * 100;

[0200] High-reading control wells: cells plus 100 nL DMSO;

[0201] Low reading control wells: wells without cells.

[0202] Table 1: Inhibitory activity of test compounds on SK-BR-3 proliferation

[0203] The results showed that the compound of the present invention has a strong inhibitory effect on the proliferation of SK-BR-3 cells. Test Example 2 Inhibition experiment of test compounds on the proliferation of MDA-MB-468 cells

[0204] 2.1 Experimental Materials

[0205] 2.2 Experimental instruments

[0206] 2.3 Test Method

[0207] (1) Cell plating: First, culture the tumor cells MDA-MB-468 in the corresponding culture medium, digest the cells with trypsin, resuspend the cells after centrifugation, count the cells, adjust the cells to the appropriate concentration, and plate them on a 384-well plate.

[0208] (2) Co-incubation of compounds and tumor cells: After cells adhered, 100 nL of diluted bioactive substance (test compound) was added to the cell culture plate using ECHO. The final DMSO concentration in the cell plate wells was 0.33%. The cells were incubated in a 37°C, 0% CO2 incubator for 72 h.

[0209] (3) After the incubation, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the wells were shaken on a rapid shaker for 2 minutes, centrifuged at 1000 rpm for 1 minute, and placed in the dark at room temperature for 30 minutes. The chemiluminescence signal value was read using an Envision instrument.

[0210] (4) Cell viability assay: IC was calculated using GraphPad Prism 8 software 50 , the IC of the compound is obtained using the following nonlinear fitting formula 50 (See Table 2):

[0211] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0212] Y: inhibition rate; X: log value of compound concentration;

[0213] Inhibition rate (%) = 100 - (reading value of compound well - reading value of low reading control well) / (reading value of high reading control well - reading value of low reading control well) * 100;

[0214] High read value control well: cells plus 100nL DMSO

[0215] Low reading control wells: wells without cells.

[0216] Table 2: Antiproliferation activity of test compounds on MDA-MB-468

[0217] The results showed that the test compound had a strong inhibitory effect on the proliferation of MDA-MB-468 cells.

[0218] Test Example 3 Inhibitory experiment of test compound on NCI-N87 cell proliferation

[0219] 3.1 Experimental Materials

[0220] 3.2 Experimental instruments

[0221] 3.3 Test Method

[0222] (1) Cell plating: First, culture the tumor cells NCI-N87 in the corresponding culture medium, digest the cells with trypsin, resuspend the cells after centrifugation, count the cells, adjust the cells to an appropriate concentration, and plate them on a 384-well plate.

[0223] (2) Co-incubation of compounds and tumor cells: After cells adhered, 100 nL of diluted bioactive substance (test compound) was added to the cell culture plate using ECHO. The final DMSO concentration in the cell plate wells was 0.33%. The cells were incubated in a 37°C, 0% CO2 incubator for 72 h.

[0224] (3) After the incubation, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the wells were shaken on a rapid shaker for 2 minutes, centrifuged at 1000 rpm for 1 minute, and placed in the dark at room temperature for 30 minutes. The chemiluminescence signal value was read using an Envision instrument.

[0225] (4) Cell viability assay: IC was calculated using GraphPad Prism 8 software 50 , the IC of the compound was obtained using the following nonlinear fitting formula: 50 (See Table 3):

[0226] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0227] Y: inhibition rate; X: log value of compound concentration;

[0228] Inhibition rate (%) = 100 - (reading value of compound well - reading value of low reading control well) / (reading value of high reading control well - reading value of low reading control well) * 100;

[0229] High-reading control wells: cells plus 100 nL DMSO;

[0230] Low reading control wells: wells without cells.

[0231] Table 3 Inhibitory activity of test compounds on NCI-N87 cell proliferation

[0232] The results showed that the test compound had a strong inhibitory effect on the proliferation of NCI-N87 cells.

[0233] The above is an exemplary description of the implementation methods of the technical solution of the present invention. It should be understood that the scope of protection of the present invention is not limited to the above implementation methods. Any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art within the spirit and principles of the present invention shall be included in the scope of protection of the claims of this application.

Claims

1. A compound of Formula I, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt, or prodrug thereof: in, Y is halogen; R1, R2, R3, R4 are the same or different and are independently selected from H, OH, CN, halogen, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 1-10 Alkoxy, halogenated C 1-10 Alkyl, halogenated C 1-10 Alkoxy, C 3-10 Cycloalkyl or halogenated C 3-10 Cycloalkyl; R 51 、R 52 The same or different, independently selected from H, OH, CN, halogen, C 1-10 Alkyl, halogenated C 1-10 Alkyl, C 3-10 Cycloalkyl or halogenated C 3-10 Cycloalkyl; Alternatively, R2 and R3 together with the carbon atoms to which they are attached form C 3-10 Carbocyclic ring or 3-10 membered heterocyclic ring; Alternatively, R3 and R4 together with the carbon atoms to which they are attached form C 3-10 Carbocyclic ring or 3-10 membered heterocyclic ring; Alternatively, R4 and R 51 Together with the carbon atoms to which they are attached, they form C 5-10 Carbocyclic ring or 5-10 membered heterocyclic ring; L1 is absent or selected from unsubstituted or optionally substituted by one, two or more selected from halogen, CN, C 1-10 Alkyl, halogenated C 1-10 Alkyl, or C 3-10 Substituents of cycloalkyl substituted C 1-10 alkylene; X is selected from CR X1 R X2 NR X3 , O or S; R X1 、R X2 and R X3 the same or different, which are absent or independently selected from H or C 1-10 alkyl; R6 is selected from H, halogen or C 1-10 alkyl; R7 is selected from OH or NH2; R 81 、R 82 The same or different, independently selected from H or C 1-10 alkyl; Or, when X is selected from CR X1 R X2 or NR X3 When R X1 、R X2 or R X3 Any one and R 81 Together with their respective connected atoms, they form C 3-10 Carbocyclic ring or 3-10 membered heterocyclic ring; R 91 、R 92 the same or different, independently selected from H, halogen, C 1-10 Alkyl, halogenated C 1-10 Alkyl or C 3-10 Cycloalkyl; Or, R 81 and R 91 Together with the carbon atoms to which they are attached, they form C 3-10 Carbocyclic ring or 3-10 membered heterocyclic ring; wherein the carbocyclic or heterocyclic ring is unsubstituted or optionally substituted by one, two or more selected from halogen, CN, C 1-10 Alkyl, C 1-10 Alkoxy, halogenated C 1-10 Alkyl, halogenated C 1-10 Alkoxy, C 3-10 Cycloalkyl or halogenated C 3-10 Substitution of cycloalkyl groups; m is selected from 0, 1, 2, 3, 4, 5 or 6.

2. The compound of formula I according to claim 1, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound, characterized in that: Y is selected from F, Cl, Br; Preferably, Y is selected from F; Preferably R1 is selected from H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 3-6 Cycloalkyl or halogenated C 3-6 Cycloalkyl; Preferably, R1 is selected from H; Preferably R2 is selected from H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 3-6 Cycloalkyl or halogenated C 3-6 Cycloalkyl; Preferably, R2 is selected from H or F; Preferably, R3 is selected from H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 3-6 Cycloalkyl or halogenated C 3-6 Cycloalkyl; Preferably, R3 is selected from H or methyl; Preferably, R4 is selected from H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 3-6 Cycloalkyl or halogenated C 3-6 Cycloalkyl; Preferably, R4 is selected from H or Cl; Preferably, R 51 、R 52 The same or different, independently selected from H or C 1-4 alkyl; Preferably, R 51 、R 52 All selected from H; Preferably, R2 and R3 together with the carbon atom to which they are attached form C 5-6 Carbocyclic or 5-6 membered heterocyclic ring; Preferably, R2 and R3 together with the carbon atoms to which they are attached form a 5-membered oxygen heterocycle (e.g. and in The position is fused with a benzene ring )； Preferably, R3 and R4 together with the carbon atoms to which they are attached form a 5-membered carbocyclic ring (e.g. and in The position is fused with a benzene ring )； Preferably, R4 and R 51 Together with the carbon atoms to which they are attached, they form C 5-7 Carbocyclic or 5-7 membered heterocyclic ring; Preferably, R4 and R 51 Together with the carbon atoms to which they are attached, they form C 5-7 Carbocyclic ring or 5-7 membered oxygen-containing heterocyclic ring; Preferably, L1 is absent or selected from C 1-4 alkylene; Preferably, L1 is absent or selected from methylene; Preferably, X is selected from CH2, NH or O; Preferably, R6 is selected from H or F; Preferably, R 81 、R 82 All selected from H; Preferably, when X is selected from CR X1 R X2 or NR X3 When R X1 、R X2 or R X3 Any one and R 81 Together with their respective connected atoms, they form C 5-6 Carbocyclic ring or 5-6 membered N-containing heterocyclic ring; Preferably, when X is selected from NR X3 When R X3 and R 81 Together with the atoms to which they are attached, they form a tetrahydropyrrole ring or a piperidine ring; Preferably, R 91 、R 92 All selected from H; Preferably, R 81 and R 91 Together with the carbon atoms to which they are attached, they form C 5-7 Carbocyclic or 5-7 membered heterocyclic ring; Preferably, R 81 and R 91 Together with the carbon atoms to which they are attached, they form a cyclopentane ring or a cyclohexane ring; Preferably, the carbocyclic or heterocyclic ring is unsubstituted or optionally substituted by one, two or more groups selected from halogen, CN, C 1-4 Alkyl (such as methyl, ethyl), C 1-4 Alkoxy (e.g. methoxy, ethoxy), halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 3-6 Cycloalkyl or halogenated C 3-6 Substitution of cycloalkyl groups; Preferably, m is selected from 0, 1, 2 or 3.

3. The compound of formula I according to claim 1 or 2, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound, characterized in that: The compound represented by formula (I) has the structure shown below: Among them, R1, R2, R3, R4, R 51 、R 52 , R7, R 81 、R 82 、R 91 、R 92 , L1, X, m are independently defined as described herein; Preferably, the compound represented by formula (I) has the structure shown below: Among them, R1, R2, R3, R4, R6, R7, R 81 、R 82 、R 91 、R 92 , L1, X, m are independently defined as described herein; X2 selected from CR X21 R X22 NR X23 , O or S; R X21 、R X22 and R X23 The same or different, independently selected from H or C 1-10 alkyl; Preferably, X2 is selected from CH2 or O; Preferably, the compound represented by formula (I) has the structure shown below: wherein R1, R2, R3, R4, R6, R7, and X are independently defined as described herein.

4. The compound of formula I according to any one of claims 1 to 3, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound, characterized in that: The structure of the compound of formula I is shown below:

5. A structural fragment D having the structure of the compound of formula I according to any one of claims 1 to 4 after dehydrogenation; Preferably, the structure of D is as follows:

6. A compound represented by Formula V, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound: L'-D (Formula V) in, L' is a linker containing a linker portion M that can react with an antibody or an antigen-binding fragment thereof, and after L'-D reacts with the antibody or an antigen-binding fragment thereof, L' forms a linker L; D has the definition described in claim 5.

7. An antibody drug conjugate of formula VI, Ab-[LD] β (Formula VI) in, Ab is an antibody or an antigen-binding fragment thereof, D has the definition described in claim 5, L is a linker connecting Ab and D, and β is selected from an integer or decimal between 1 and 10.

8. A method for preparing the compound according to any one of claims 1 to 4, comprising the following steps: Among them, R1, R2, R3, R4, R 51 、R 52 、L1、X、R6、R7、R 81 、R 82 、R 91 、R 92 and m are independently defined as described herein; Z is a leaving group (eg, F, Cl, Br).

9. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the compound of formula I according to any one of claims 1 to 4 or the compound of formula V according to claim 6, or a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt, or prodrug thereof; Preferably, the pharmaceutical composition comprises a therapeutically effective amount of the antibody-drug conjugate of claim VI according to claim 7; Preferably, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

10. Use of at least one of the compound of formula I according to any one of claims 1 to 4, the compound of formula V according to claim 6, or the antibody-drug conjugate of formula VI according to claim 7, their racemates, stereoisomers, tautomers, solvates, polymorphs, pharmaceutically acceptable salts, or prodrugs thereof, or the pharmaceutical composition according to claim 9 in the preparation of a topoisomerase I inhibitor and / or in the preparation of a medicament for preventing or treating a disease or condition associated with topoisomerase I; Preferably, the disease or condition is a tumor, and the tumor includes breast cancer, gastric cancer, lung cancer, colorectal cancer, large intestine cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal cancer, melanoma or leukemia.

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