Camptothecin compound and use thereof

Abstract

The present invention relates to a compound represented by formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope labeled compound, metabolite or prodrug thereof, a pharmaceutical composition comprising same, and a use thereof in preventing or treating cancer and / or tumor and related diseases. The compound has good activity and a more stable lactone ring, also has both a low efflux rate and better safety, and has obvious druggability potential.

Description

A camptothecin-type compound and its uses Technical Field

[0001] This disclosure relates to a novel camptothecin analogue, pharmaceutical compositions comprising the camptothecin, and their use in the preparation of pharmaceuticals. Background Technology

[0002] DNA topoisomerases are a class of enzymes found in the cell nucleus that catalyze the breaking and binding of DNA strands, thereby controlling the topological state of DNA. Topoisomerases participate in the regulation of supercoiled template structures. DNA topoisomerase I catalyzes changes in the topological isomerism of DNA replication by forming a brief single-strand cleavage-binding cycle. Camptothecin, a pentacyclic quinoline alkaloid originally isolated from the bark and stems of the Chinese camptotheca, is one of the important lead compounds in the development of anticancer drugs. Its structure is a planar pentacyclic system, including a quinoline ring AB, a pyrrole ring C, a pyridone ring D, and an α-hydroxylactone ring E, with the 20th chiral carbon in the S configuration serving as the active site of camptothecin. Camptothecin is a Topo I inhibitor; it forms a stable Topo I-DNA-CPT ternary complex by binding to the Topo I-DNA binary complex, inhibiting DNA rejoining, causing DNA damage, and ultimately leading to cell death. To address the drawbacks of camptothecin, such as poor solubility, stability, and high toxicity, camptothecin derivatives can increase water solubility and improve drug properties by introducing water-soluble groups or preparing prodrugs. Several camptothecin derivatives, such as topotecan and irinotecan, have already been approved for marketing and are used to treat solid tumors. Developed by Daiichi Sankyo, the irinotecan derivative Dxd is a highly effective Topo I inhibitor. As an effective payload for antibody-drug conjugates (ADCs), it selectively kills tumor cells through antibody targeting. Compared to traditional chemotherapy drugs, ADCs can more precisely kill tumor cells while reducing toxicity to normal tissues. Several ADC drugs using Dxd as the payload have been approved for marketing or entered clinical trials, demonstrating excellent efficacy and manageable safety in solid tumors such as breast cancer and non-small cell lung cancer. However, Dxd is a substrate for the transporter P-gp and BCRP, making it easily effluxed by tumor cells, reducing its efficacy. Meanwhile, Dxd exhibits poor sensitivity in killing gastrointestinal tumor cells, which may be associated with the development of interstitial lung disease. Therefore, developing novel camptothecin derivatives that are sensitive to the gastrointestinal tract and have low efflux as effective payloads for ADCs is of great significance and has promising application prospects.

[0003] Invention Overview

[0004] Through extensive research, the inventors have surprisingly discovered that the camptothecin derivative described in this application has good activity, its lactone ring is more stable, and it has a low efflux rate and better safety, showing significant drug development potential.

[0005] One aspect of the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, metabolite, isotopically labeled compound, or prodrug thereof.

[0006] in:

[0007] R 1 and R 2 Independently selected from hydroxyl, halogen, -C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, -C 1-3 Alkyl-NR a R b or -NR a R b Or, R 1 R 2 It forms a 5-6 membered heterocyclic alkyl group with adjacent C atoms;

[0008] R 3 Selected from H, C 1-3 Alkyl or C 1-3 Halogenated alkyl groups;

[0009] R 4 Selected from C 1-6 Haloalkyl, C 1-6 Alkyl or C 3-10 cycloalkyl, and the R 4 Further by R 4A replace;

[0010] R 4A Selected from hydroxyl, thiol or -NR 4A-1 -(CH2) m -R 4A-2 And the R 4A It can be optionally substituted by halogens at substituted sites;

[0011] R 4A-1 Selected from H, C 1-3 Alkyl, C 1-3 Halogenated alkyl or C 1-3 Alkoxy;

[0012] R 4A-2 Selected from H, halogens, =O, CN, -S(=O)2-C 1-3Alkyl group, -S(=O)2-C 4-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heterocycloalkyl, C 6-10 Aryl or 5-7 heteroaryl;

[0013] Or, R 3 R 4 It forms a 5-10 membered cycloalkyl group with adjacent C atoms, wherein the 5-10 membered cycloalkyl group is optionally surrounded by one or more R atoms. x replace;

[0014] R x Independently selected from halogens, NR c R d hydroxyl group, C 1-3 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-3 Halogenated alkyl or C 1-3 Alkoxy;

[0015] R a and R b Independently selected from H and C 1-3 Alkyl or C 1-3 Halogenated alkyl groups;

[0016] R c and R d Independently selected from H or -COCH3;

[0017] n is 0 or 1;

[0018] m can be 0, 1, 2, 3, or 4;

[0019] When n is 0, and R 3 R 4 When forming a 5-10 membered cycloalkyl group with an adjacent C atom, the 5-10 membered cycloalkyl group is cyclohexyl, and the 5-10 membered cycloalkyl group is substituted with -NHAc, R 1 and R 2 Independently selected from halogens; or, the 5-10 membered cycloalkyl group is cyclohexyl, and the 5-10 membered cycloalkyl group is optionally substituted with a halogen, R 1 Hydroxyl, halogen, C 1-6 Halogenated alkyl, -C 1-3 Alkyl-NR a R b Or C 1-6 Alkoxy, R 2 It is a halogen; or, the 5-10 membered cycloalkyl group is cyclohexyl, R 1 For hydroxyl group, R 2It is a halogen; or, the 5-10 membered cycloalkyl group is a 5-10 membered bridged cycloalkyl group, and the 5-10 membered cycloalkyl group is optionally bonded by one or more R groups. x Substitution; or, the 5-10 membered cycloalkyl group is a partially unsaturated 5-10 membered cycloalkyl group, and the 5-10 membered cycloalkyl group is optionally replaced by one or more R groups. x replace;

[0020] When n is 1, and R 3 R 4 When R forms a 5-10 membered cycloalkyl group with adjacent C atoms, 1 and R 2 Independently selected from hydroxyl, halogen, -C 1-6 Alkyl, C 1-6 Halogenated alkyl or C 1-6 alkoxy, or, R 1 R 2 It forms a 5-6 membered heterocyclic alkyl group with adjacent C atoms;

[0021] When R 4 C 1-6 When alkyl, n is 1, and R 1 Selected from hydroxyl, halogen, -C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 alkoxy or -C 1- 3alkyl-NR a R b .

[0022] Another aspect of the invention provides a pharmaceutical composition comprising a preventive or therapeutically effective amount of the compound of the invention or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug, and one or more pharmaceutically acceptable carriers.

[0023] Another aspect of the invention provides the use of the compounds of the invention or pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs, or pharmaceutical compositions of the invention in the preparation of medicaments for the prevention or treatment of cancer and / or tumors and related conditions.

[0024] Another aspect of the present invention provides compounds of the present invention or pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs of the present invention, or pharmaceutical compositions of the present invention, for the prevention or treatment of cancer and / or tumors and related conditions.

[0025] Another aspect of the present invention provides a method for preventing or treating cancer and / or tumors and related conditions, the method comprising administering to an individual in need an effective amount of the compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug or pharmaceutical composition of the present invention.

[0026] Invention Details

[0027] definition

[0028] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to technical terms herein refer to techniques commonly understood in the art, including variations or equivalent substitutions of techniques that are obvious to one of ordinary skill in the art. While it is believed that the following terms will be well understood by one of ordinary skill in the art, the following definitions are set forth to better explain the invention.

[0029] The terms “including,” “comprising,” “having,” “containing,” or “involving,” and their other variations herein, are inclusive or open-ended and do not exclude other unlisted elements or method steps.

[0030] As used herein, the term "alkyl" is defined as a linear or branched saturated aliphatic hydrocarbon. In some embodiments, the alkyl group has 1 to 12, for example, 1 to 6 carbon atoms. For example, as used herein, the term "Cw alkyl" refers to a linear or branched group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, or n-hexyl) of 1 to 6 carbon atoms, optionally substituted with one or more (e.g., 1 to 3) suitable substituents such as halogens (in which case the group is referred to as "haloalkyl") (e.g., CH2F, CHF2, CF3, CCl3, C2F5, C2C15, CH2CF3, CH2Cl, or -CH2CH2CF3, etc.). The term "CM alkyl" refers to a linear or branched aliphatic hydrocarbon chain (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl) of 1 to 4 carbon atoms.

[0031] As used herein, the term "alkenyl" refers to a linear or branched monovalent hydrocarbon group containing a double bond; for example, "CM alkenyl" is an alkenyl group having 2-6 carbon atoms. The alkenyl group is, for example, vinyl, 1-propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl, and 4-methyl-3-pentenyl. When the compounds of the present invention contain an alkenyl group, the compounds may exist in pure E (iso-side), pure Z (iso-side), or any mixture thereof.

[0032] As used herein, the term "alkynyl" refers to a monovalent hydrocarbon group containing one or more triple bonds; for example, "CM alkynyl" is an alkynyl group having 2 to 6 carbon atoms. Examples of alkynyl groups include ethynyl or propynyl.

[0033] As used herein, the term "cycloalkyl group" refers to a saturated or unsaturated (i.e., having one or more double and / or triple bonds within the ring) monocyclic or polycyclic hydrocarbon ring having, for example, 3 to 10 (suitably 3 to 8, more preferably 3 to 6) cyclic carbon atoms, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclohexenyl, etc., wherein the polycyclic hydrocarbon ring includes spirocyclic, bridged, or fused rings.

[0034] As used herein, the term "cycloalkyl" should be interpreted broadly to include saturated and partially unsaturated (i.e., having one or more double and / or triple bonds within the ring) monocyclic or polycyclic hydrocarbon rings. For example, when describing compounds of formula (I) of the present invention in the preceding and following text, the expression "the 5-10 membered cycloalkyl is a partially unsaturated 5-10 membered cycloalkyl" is used. Those skilled in the art will understand that when "xy membered cycloalkyl" is mentioned herein, it actually refers to "C x-y "Cycloalkyl". In other words, in this application, "xy-membered cycloalkyl" and "C" are used interchangeably. x- y "Cycloalkyl" can be used interchangeably.

[0035] As used herein, the term "heterocyclic group" refers to a cyclic group having, for example, 3 to 10 (suitably 3 to 8, more preferably 3 to 6) ring atoms, wherein at least one ring atom is a heteroatom selected from N, O, S, and P, and the remaining ring atoms are C, and is either saturated (heterocyclic alkyl) or partially unsaturated (i.e., having one or more double and / or triple bonds within the ring). For example, "3 to 10-membered heterocyclic group" is a saturated or partially unsaturated heterocyclic group having 2 to 9 (e.g., 2, 3, 4, 5, 6, 7, 8, or 9) ring carbon atoms and one or more (e.g., 1, 2, 3, or 4) heteroatoms independently selected from N, O, and S. Examples of heterocyclic groups include, but are not limited to: ethylene oxide, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuranyl, dioxolinyl, pyrrolylyl, pyrrolidoneyl, imidazoalkyl, pyrazolyl, pyrrolinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, or trithianyl. The groups also encompass bicyclic systems, including spirocyclic, fused, or bridged systems (such as 8-azaspiro[4.5]decane, 3,9-diazaspiro[5.5]undecane, 2-azabicyclo[2.2.2]octane, etc.). The heterocyclic group may optionally be substituted with one or more (e.g., 1, 2, 3, or 4) suitable substituents.

[0036] As used herein, the term "aryl" refers to a monocyclic or polycyclic aromatic group with a conjugated electron system. For example, as used herein, the term "C WD "Aryl" refers to an aromatic group containing 6 to 10 carbon atoms, such as phenyl or naphthyl. The aryl group may optionally be replaced by one or more (such as 1 to 3) suitable substituents (e.g., halogen, -OH, -CN, -NO2, Cw alkyl, etc.).

[0037] As used herein, the term "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic aromatic ring system. For example, "5-14 membered heteroaryl" means having 5, 6, 8, 9, 10, 11, 12, 13, or 14 ring atoms, particularly 1, or 2, or 3, or 4, or 5, or 6, or 9, or 10 carbon atoms, and containing at least one heteroatom that may be the same or different (the heteroatom being, for example, oxygen, nitrogen, or sulfur), and additionally, in each case, may be benzofused. Examples of "heteroaryl" include, but are not limited to: thiophene, furanyl, pyrrole, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiazolyl, etc., and their benzo[derivatives]; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc., and their benzo[derivatives].

[0038] As used herein, the term “halogenated” or “halogenated” is defined as including F, Cl, Br, or I.

[0039] The term "R" used in this application 1 R 2 "Forming a 5-6 membered heterocyclic alkyl group with adjacent C atoms", "The R 1 R 2 Composed of adjacent C atoms ”, and “R” 3 R 4 The phrase "forming a 5-10 membered cycloalkyl group with adjacent C atoms" and similar expressions refer to R. 1 and R 2 The connected ring C atom, or R 3 and R 4 The C atoms of the attached ring. In other words, R 1 and R 2 Together with the ring C atoms to which they are attached, they form the indicated cyclic group, which is fused with the ring containing the said ring C atoms; and R 3 and R 4 Together with the ring C atoms to which they are attached, they form the indicated cyclic group, which is fused with the ring containing the ring C atoms.

[0040] The term "substitution" refers to the selective replacement of one or more (e.g., one, two, three, or four) hydrogen atoms on a specified atom by a designated group, provided that the substitution does not exceed the normal valence of the specified atom in the present case and that the substitution forms a stable compound. Combinations of substituents and / or variables are permitted only if such combinations form a stable compound.

[0041] If a substituent is described as “optionally substituted,” then the substituent may be (1) unsubstituted or (2) substituted. If the carbon of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the carbon (to the extent that any hydrogens are present) may be substituted individually and / or together with independently selected optional substituents. If the nitrogen of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the nitrogen (to the extent that any hydrogens are present) may each be substituted with independently selected optional substituents.

[0042] If a substituent is described as being “independently selected” from a group, then each substituent is selected independently of the others. Therefore, each substituent may be the same as or different from another (other) substituent.

[0043] As used herein, the term "one or more" means one or more under reasonable conditions, such as two, three, four, five, or ten.

[0044] Unless otherwise specified, as used herein, the connection point of a substituent may be located at any suitable position of the substituent.

[0045] This invention also includes all pharmaceutically acceptable isotopically labeled compounds that are identical to the compounds of this invention, except that one or more atoms are replaced by atoms having the same atomic number but with an atomic mass or mass number different from the dominant atomic mass or mass number in nature. Examples of isotopes suitable for inclusion in the compounds of this invention include (but are not limited to) isotopes of hydrogen (e.g., deuterium). 2 H), tritium ( 3 H); carbon isotopes (e.g., ... 11 C 13 C and 14 C); isotopes of chlorine (e.g.) 36 Cl); isotopes of fluorine (e.g., Cl); 18 F); isotopes of iodine (e.g., F); 123 I and 125 I); nitrogen isotopes (e.g.) 13 N and 15 N); isotopes of oxygen (e.g., N); 15 O、 17 O and 18 O); isotopes of phosphorus (e.g., O); phosphorus isotopes ... 32 P); and isotopes of sulfur (e.g. 35 S). Certain isotope-labeled compounds of the present invention (e.g., those doped with radioactive isotopes) can be used in drug and / or substrate tissue distribution studies (e.g., analysis). Radioactive isotope tritium (i.e. 3 H) and carbon-14 (i.e. 14 C) It is particularly suitable for this purpose due to its ease of incorporation and detection. Using positron-emitting isotopes (e.g.) 11 C 18 F, 15 O and 13 N) substitution can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of the present invention can be prepared by methods similar to those described in the accompanying routes and / or examples and preparations, using a suitable isotopically labeled reagent instead of the previously used unlabeled reagent. Pharmaceutically acceptable solvates of the present invention include those in which the crystallization solvent can be isotopically substituted, for example, D. 2 O, acetone or DMSO-*.

[0046] The term "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In compounds having one or more (e.g., one, two, three, or four) asymmetric centers, racemic mixtures, single enantiomers, diastereomer mixtures, and individual diastereomers can be produced. Specific individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of the present invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers).

[0047] Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. It should be understood that the scope of this application covers all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

[0048] Solid lines may be used in this article. solid wedge Or virtual wedge The chemical bonds of the compounds of the present invention are depicted. Solid lines are used to depict bonds to asymmetric carbon atoms to indicate all possible stereoisomers (e.g., specific enantiomers, racemic mixtures, etc.) at that carbon atom. Solid or dashed wedges are used to depict bonds to asymmetric carbon atoms to indicate the presence of the indicated stereoisomers. When present in racemic mixtures, solid and dashed wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise specified, the compounds of the present invention are intended to exist as stereoisomers (including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, trans-blocking isomers, and mixtures thereof). The compounds of the present invention may exhibit more than one type of isomerism and may consist of mixtures thereof (e.g., racemic mixtures and diastereomer pairs).

[0049] This invention encompasses all possible crystalline forms or polymorphs of the compounds of this invention, which may be a single polymorph or a mixture of more than one polymorph in any proportion. It should also be understood that certain compounds of this invention may exist in a free form for therapeutic purposes, or, where appropriate, in their pharmaceutically acceptable derivative forms. In this invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, N-oxides, metabolites, or prodrugs, which, upon administration to a patient in need, can directly or indirectly provide the compounds of this invention or their metabolites or residues. Therefore, when referring to "compounds of this invention" herein, it is also intended to cover the various derivative forms of the compounds described above.

[0050] Pharmaceutically acceptable salts of the compounds of the present invention include their acid addition salts and base addition salts. Suitable acid addition salts are formed by acids that form pharmaceutically acceptable salts. Suitable base addition salts are formed by bases that form pharmaceutically acceptable salts. A review of suitable salts can be found in Stahl and Wermuth's "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the present invention are known to those skilled in the art.

[0051] As used herein, the term "ester" means an ester derived from the various general formula compounds of this application, including physiologically hydrolyzable esters (the compounds of the present invention that can be hydrolyzed under physiological conditions to release free acids or alcohols). The compounds of the present invention may themselves also be esters.

[0052] The compounds of the present invention can exist as solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent, particularly, for example, water, methanol, or ethanol, as a structural element of the lattice of the compound. The amount of the polar solvent, particularly water, can be stoichiometric or non-stoichiometric.

[0053] Those skilled in the art will understand that not all nitrogen-containing heterocycles can form oxides because nitrogen requires available lone pairs of electrons to be oxidized. Those skilled in the art will identify nitrogen-containing heterocycles that can form oxides. Those skilled in the art will also recognize that tertiary amines can form N-oxides. Synthetic methods for preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art, including the oxidation of heterocycles and tertiary amines with peroxyacids such as peracetic acid and m-chloroperoxybenzoic acid (MCPBA), hydrogen peroxide, alkyl peroxides such as tert-butyl peroxide, sodium perborate, and dioxiranes such as dimethyldioxirane. These methods for preparing W-oxides have been extensively described and reviewed in the literature, see, for example: T.L. Gilchrist, Comprehensive Organic Synthesis, vol. 7, pp. 748-750; A.R. Katritzky and A.J. Boulton, Eds., Academic Press; and G.W. H. Heeseman and E.S. G. Wierstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp. 390-392, A.R. Katritzky and A.J. Boulton, Eds., Academic Press.

[0054] The scope of this invention also includes metabolites of the compounds of this invention, i.e., substances formed in the body when the compounds of this invention are administered. Such products can be generated, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc., of the administered compound. Therefore, this invention includes metabolites of the compounds of this invention, including compounds obtained by methods that expose the compounds of this invention to mammals for a time sufficient to produce their metabolites.

[0055] This invention further includes, within its scope, prodrugs of the compounds of the invention, which are certain derivatives of the compounds of the invention that may themselves have little or no pharmacological activity, which, when administered to or onto the body, can be converted, for example, by hydrolysis and cleavage into the compounds of the invention having the desired activity. Typically, such prodrugs are functional group derivatives of the compounds that are readily converted in vivo into the compounds with the desired therapeutic activity. Further information regarding the use of prodrugs can be found in “Pro-drugs as Novel Delivery Systems,” Vol. 14, ACS Symposium Series (T. Higuchi and V. Stella). The prodrugs of the invention can be prepared, for example, by replacing suitable functional groups present in the compounds of the invention with certain portions known to those skilled in the art as “pro-moiety” (e.g., “Design of Prodmgs,” H. Bundgaard (Elsevier, 1985)).

[0056] This invention also covers compounds of the invention containing protecting groups. In any process of preparing the compounds of the invention, protection of sensitive or reactive groups on any relevant molecule may be necessary and / or desired, thereby forming a form of chemical protection for the compounds of the invention. This can be achieved by conventional protecting groups, for example, those described in T.W. Greene & P. ​​G. W. M. Uts, *Protective Groups in Organic Synthesis*, John Wiley & Sons, 1991, which are incorporated herein by reference. Protecting groups can be removed at appropriate subsequent stages using methods known in the art.

[0057] The term “about” means within ±10% of the stated value, preferably within ±5%, and more preferably within ±2%.

[0058] compound

[0059] One aspect of the present invention provides compounds of formula (I), or pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, N-oxides, metabolites, isotopically labeled compounds, or prodrugs thereof:

[0060] in:

[0061] R 1 and R 2 Independently selected from hydroxyl, halogen, -C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, -C 1-3 Alkyl-NR a R b or -NR a R b Or, R 1 R 2 It forms a 5-6 membered heterocyclic alkyl group with adjacent C atoms;

[0062] R 3 Selected from H, C 1-3 Alkyl or C 1-3 Halogenated alkyl groups;

[0063] R 4 Selected from C 1-6 Haloalkyl, C 1-6 Alkyl or C 3-10 cycloalkyl, and the R 4 Further by R 4A replace;

[0064] R 4A Selected from hydroxyl, thiol or -NR 4A-1 -(CH2) m -R 4A-2 And the R 4A It can be optionally substituted by halogens at substituted sites;

[0065] R 4A-1 Selected from H, C 1-3 Alkyl, C 1-3 Halogenated alkyl or C 1-3 Alkoxy;

[0066] R 4A-2 Selected from H, halogens, =O, CN, -S(=O)2-C 1-3 Alkyl group, -S(=O)2-C 4-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heterocycloalkyl, C 6-10 Aryl or 5-7 heteroaryl;

[0067] Or, R 3 R 4 It forms a 5-10 membered cycloalkyl group with adjacent C atoms, and the 5-10 membered cycloalkyl group is optionally surrounded by one or more R atoms.x replace;

[0068] R x Independently selected from halogens, NR c R d hydroxyl group, C 1-3 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-3 Halogenated alkyl or C 1-3 Alkoxy;

[0069] R a and R b Independently selected from H and C 1-3 Alkyl or C 1-3 Halogenated alkyl groups;

[0070] R c and R d Independently selected from H or -COCH3;

[0071] n is 0 or 1;

[0072] m can be 0, 1, 2, 3, or 4;

[0073] When n is 0, and R 3 R 4 When forming a 5-10 membered cycloalkyl group with an adjacent C atom, the 5-10 membered cycloalkyl group is cyclohexyl, and the 5-10 membered cycloalkyl group is substituted with -NHAc, R 1 and R 2 Independently selected from halogens; or, the 5-10 membered cycloalkyl group is cyclohexyl, and the 5-10 membered cycloalkyl group is optionally substituted with a halogen, R 1 Hydroxyl, halogen, C 1-6 Halogenated alkyl, -C 1-3 Alkyl-NR a R b Or C 1-6 Alkoxy, R 2 It is a halogen; or, the 5-10 membered cycloalkyl group is cyclohexyl, R 1 For hydroxyl group, R 2 It is a halogen; or, the 5-10 membered cycloalkyl group is a 5-10 membered bridged cycloalkyl group, and the 5-10 membered cycloalkyl group is optionally bonded by one or more R groups. x Substitution; or, the 5-10 membered cycloalkyl group is a partially unsaturated 5-10 membered cycloalkyl group, and the 5-10 membered cycloalkyl group is optionally replaced by one or more R groups. x replace;

[0074] When n is 1, and R 3 R 4 When R forms a 5-10 membered cycloalkyl group with adjacent C atoms,1 and R 2 Independently selected from hydroxyl, halogen, -C 1-6 Alkyl, C 1-6 Halogenated alkyl or C 1-6 alkoxy, or, R 1 R 2 It forms a 5-6 membered heterocyclic alkyl group with adjacent C atoms;

[0075] When R 4 C 1-6 When alkyl, n is 1 and R 1 Selected from hydroxyl, halogen, -C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 alkoxy or -C 1-3 Alkyl-NR a R b .

[0076] In some embodiments, this disclosure provides compounds of formula (I) as described above, wherein R x Independently selected from halogens, NR c R d C 1-3 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-3 Halogenated alkyl or C 1-3 Alkyl group.

[0077] In some embodiments, this disclosure provides compounds of formula (I) as described above, wherein:

[0078] R 4 Selected from C 1-6 Haloalkyl, C 1-6 Alkyl or C 3-10 cycloalkyl, and the R 4 Further by R 4A replace;

[0079] R 4A Selected from hydroxyl or -NR 4A-1 -(CH2) m -R 4A-2 And the R 4A It can be optionally substituted by halogens at substituted sites;

[0080] R 4A-2 Selected from H, halogens, =O, CN, -S(=O)2-C 1-3 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heterocycloalkyl, C 6-10 Aryl or 5-7 heteroaryl compounds.

[0081] In some embodiments, this disclosure provides compounds of formula (I) as described above, wherein R 4A-1 For H.

[0082] In some embodiments, this disclosure provides compounds of formula (I) as described above, wherein

[0083] R 4 Selected from C 1-6 Haloalkyl, C 1-6 Alkyl or C 3-10 cycloalkyl, the R 4 Further replaced by hydroxyl groups, and R c and R d H stands for H independently.

[0084] In some embodiments, this disclosure provides compounds of formula (I) as described above, wherein the compounds of formula (I) are compounds of formula (I-1):

[0085] in:

[0086] R 1 Selected from hydroxyl, halogen, C 1-6 Halogenated alkyl, -C 1-3 Alkyl-NR a R b Or C 1-6 Alkoxy;

[0087] R 2 Selected from halogens;

[0088] R L Optionally selected from halogens, preferably halogens, more preferably F; and

[0089] m is 2.

[0090] In some preferred embodiments, R 1 The derivative is selected from hydroxyl, F, Cl, Br, I, -CF3, -CHF2, -CH2F, -CF2CH3, -CF2CH2CH3, -CH2CF3, -CH2CF2CH3, -CH2CH2CF3, -CH2-N(CH3)2, -CH2-NH2, methoxy, ethoxy, or propoxy, preferably hydroxyl, F, -CH2-N(CH3)2, -CH2-NH2, or methoxy.

[0091] In some embodiments, this disclosure provides compounds of formula (I) as described above, wherein the compounds of formula (I) are compounds of formula (I-2):

[0092] in:

[0093] R 1 and R 2 Independently selected from hydroxyl, halogen, C 1-6 Halogenated alkyl, -C 1-3 Alkyl-NR a R b -NR a R b or C 1-6 Alkoxy;

[0094] R L Selected from halogens, -OH, C 1-3 Alkyl or C 1-3 Halogenated alkyl groups; and

[0095] m can be 0, 1, 2, or 3.

[0096] In some preferred embodiments, R 1 and R 2 The radical is independently selected from hydroxyl, F, Cl, Br, I, -CF3, -CHF2, -CH2F, -CF2CH3, -CF2CH2CH3, -CH2CF3, -CH2CF2CH3, -CH2CH2CF3, -CH2-N(CH3)2, -NH2, methoxy, ethoxy, or propoxy, preferably hydroxyl, F, or NH2. In a more preferred embodiment, R 1 It is a hydroxyl group, F or NH2, and R 2 It is F.

[0097] In some embodiments, this disclosure provides compounds of formula (I) as described above, wherein the compounds of formula (I) are compounds of formula (I-3):

[0098] Where: R 1 and R 2 Independently selected from halogens; R L Selected from -NHAc; and m is 1.

[0099] In some implementation schemes, R 1 and R 2 The component is independently selected from F, Cl, Br, or I, preferably F or Cl. In a more preferred embodiment, R... 1 It is F or Cl, and R 2 It is F.

[0100] In some embodiments, this disclosure provides compounds of formula (I) as described above, wherein the compounds of formula (I) are compounds of formula (I-4):

[0101] in:

[0102] R3 Selected from H;

[0103] R 4 Selected from C 1-6 Halogenated alkyl or C 3-10 cycloalkyl, and the R 4 Further by R 4A replace.

[0104] In some preferred embodiments, the R 4A Selected from hydroxyl or -NH-(CH2) m -R 4A-2 And the R 4A It can be optionally substituted with halogens at sites where it can be replaced.

[0105] In some preferred embodiments, R 3 Selected from H, R 4 Selected from C 1-6 Halogenated alkyl groups, and the R 4 Further converted to hydroxyl groups or -NH-(CH2) m -R 4A-2 replace.

[0106] In some preferred embodiments, R 4A-2 Selected from H, halogens, CN, -S(=O)2-C 1-3 Alkyl or C 3-6 Cycloalkyl, and m is 0, 1, 2, or 3. In some preferred embodiments, R 4A-2 Selected from H, F, Cl, Br, I, CN, -S(=O)2-CH3 or C 3-6 cycloalkyl, wherein the C 3-6 The cycloalkyl group can be monocyclic or bridged, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[1.1.1]pentane, or bicyclo[2.1.1]hexane. In some preferred embodiments, m is 0, 1, or 2.

[0107] In some preferred embodiments, R 3 Selected from H, R 4 Selected from C 1-6 Halogenated alkyl or C 3-10 cycloalkyl, and the R 4 It is further replaced by hydroxyl groups.

[0108] In some preferred embodiments, R 3 Selected from H, R 4 Selected from C 3-10 cycloalkyl, and the R 4 It is further replaced by hydroxyl groups.

[0109] In some preferred embodiments, the R 4 Selected from -CF2OH, -CF2CH2OH, -CF2CH2CH2OH, -CH2CF2OH, -CH2CF2CH2OH, -CH2CH2CF2OH, Cyclobutyl, cyclopentyl, cyclohexyl Preferred CF₂OH, -CF₂CH₂OH, -CF₂CH₂CH₂OH, -CH₂CF₂OH, -CH₂CF₂CH₂OH, -CH₂CH₂CF₂OH Cyclobutyl, cyclopentyl, cyclohexyl More Or cyclohexyl.

[0110] In some preferred embodiments, the R 4 Selected from -CF2OH, -CF2CH2OH, -CF2CH2CH2OH, -CH2CF2OH, -CH2CF2CH2OH, -CH2CH2CF2OH, Cyclobutyl, cyclopentyl, or cyclohexyl, preferably Or cyclohexyl.

[0111] In some implementations, the R 4 It can also be a hydroxylated cyclobutyl, a hydroxylated cyclopentyl, or a hydroxylated cyclohexyl, for example...

[0112] In some preferred embodiments, the R 1 R 2 It forms a 5-6 membered heterocyclic alkyl group with adjacent C atoms, preferably the R group. 1 R 2 Composed of adjacent C atoms

[0113] In some other preferred embodiments, the R 1 Selected from C 1-3 Alkyl or C 1-3 Alkyl group.

[0114] In some preferred embodiments, the R 1 Selected from C 1-3 Alkyl group, preferably methyl group.

[0115] In some preferred embodiments, the R 1 Selected from C 1-3 Alkyl groups, preferably methoxy groups.

[0116] In some preferred embodiments, the R 2Selected from F, Cl, Br or I, with F being preferred.

[0117] In some embodiments, this disclosure provides compounds of formula (I) as described above, wherein the compounds of formula (I) are compounds of formulas (I-5):

[0118] Where R 4 Selected from C 1-6 Halogenated alkyl or C 1-6 Alkyl group, and the R 4 It is further replaced by hydroxyl groups.

[0119] In some preferred embodiments, the R 4 Selected from -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, -CF2OH, -CF2CH2OH, -CF2CH2CH2OH, -CH2CF2OH, -CH2CF2CH2OH, -CH2CH2CF2OH, Preferred

[0120] In some preferred embodiments, the R 1 R 2 It forms a 5-6 membered heterocyclic alkyl group with adjacent C atoms, preferably the R group. 1 R 2 Composed of adjacent C atoms

[0121] In some embodiments, this disclosure provides compounds of formula (I) as described above, wherein the compounds of formula (I) are compounds of formula (I-6):

[0122] in:

[0123] R 1 and R 2 Independently selected from hydroxyl, halogen, -C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, -C 1-3 Alkyl-NR a R b , or -NR a R b Or, R 1 R 2 It forms a 5-6 membered heterocyclic alkyl group with adjacent C atoms;

[0124] R L Selected from halogens, NR c R d -OH, C 1-3Alkyl, C 1-3 Alkoxy or C 1-3 Halogenated alkyl groups; and

[0125] m can be 0, 1, 2, or 3.

[0126] In some preferred embodiments, R L Selected from halogens, NR c R d Or C 1-3 Alkyl groups, preferably F, Cl, Br, I, methyl or NH2, more preferably F, methyl or NH2.

[0127] In some other preferred embodiments, R L Selected from halogens, -OH, C 1-3 Alkyl, C 1-3 Alkoxy or C 1-3 Halogenated alkyl groups, preferably halogenated or C24-hydroxyl groups. 1-3 Alkyl, more preferably F, Cl, Br, I or methyl, more preferably F or methyl.

[0128] In some preferred embodiments, m is 2 or 3.

[0129] In some preferred embodiments, R 1 Hydroxyl group, halogen, -C 1-3 Alkyl, or -NR a R b Preferably, it contains hydroxyl, F, Cl, Br, I, methyl or NH2, more preferably methyl or NH2.

[0130] In some preferred embodiments, R 2 The halogen is preferred, with F, Cl, Br or I being more preferred, and F being even more preferred.

[0131] In some embodiments, this disclosure provides compounds of formula (I) as described above, wherein the compounds of formula (I) are compounds of formula (I-7):

[0132] in:

[0133] R 1 and R 2 Independently selected from hydroxyl, halogen, -C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, -C 1-3 Alkyl-NR a R b , or -NR a R b Or, R 1 R 2 It forms a 5-6 membered heterocyclic alkyl group with adjacent C atoms;

[0134] R L Selected from halogens, -OH, C 1-3 Alkyl, C 1-3 Alkoxy or C 1-3 Halogenated alkyl groups; and

[0135] m can be 0, 1, 2, or 3.

[0136] In some preferred embodiments, R L Selected from halogen or C 1-3 Alkyl group, preferably F, Cl, Br, I or methyl, more preferably F or methyl.

[0137] In some preferred embodiments, m is 2 or 3.

[0138] In some preferred embodiments, R 1 Hydroxyl group, halogen, -C 1-3 Alkyl, or -NR a R b Preferably, it contains hydroxyl, F, Cl, Br, I, methyl or NH2, more preferably methyl or NH2.

[0139] In some preferred embodiments, R 2 The halogen is preferred, with F, Cl, Br or I being more preferred, and F being even more preferred.

[0140] In some embodiments, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, metabolite, isotopically labeled compound, or prodrug thereof, wherein the compound is selected from:

[0141] Pharmaceutical compositions and treatment methods

[0142] In some embodiments, the present invention provides pharmaceutical compositions comprising a preventatively or therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug, and one or more pharmaceutically acceptable carriers. The pharmaceutical composition is preferably a solid, semi-solid, liquid, or gaseous formulation. In some embodiments, the pharmaceutical composition may further comprise one or more other therapeutic agents. In preferred embodiments, the pharmaceutical composition is preferably administered orally, intravenously, intraarterially, subcutaneously, intraperitoneally, intramuscularly, or transdermally.

[0143] In some embodiments, the present invention provides the use of the compounds of the present invention or pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs, or pharmaceutical compositions of the present invention in the preparation of medicaments for the prevention or treatment of cancer and / or tumors and related conditions.

[0144] In some embodiments, the present invention provides compounds of the present invention or pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs of the present invention, or pharmaceutical compositions of the present invention, for the prevention or treatment of cancer and / or tumors and related conditions.

[0145] In some embodiments, the present invention provides a method for preventing or treating cancer and / or tumors and related conditions, the method comprising administering to an individual in need an effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug of the present invention, or a pharmaceutical composition of the present invention.

[0146] In some embodiments, the cancers and / or tumors and related conditions are preferably selected from: cancers occurring in the esophagus, stomach, intestines, rectum, mouth, pharynx, larynx, lungs, colon, breast, uterus, endometrium, ovary, prostate, testes, bladder, kidneys, liver, pancreas, bone, connective tissue, skin, eyes, brain and central nervous system, as well as thyroid cancer, leukemia, Hodgkin's disease, lymphoma and myeloma.

[0147] In this invention, "pharmaceutically acceptable carrier" refers to a diluent, excipient, vehicle, or medium that is administered co-administered with a therapeutic agent and is suitable, to the extent of reasonable medical judgment, for contact with human and / or other animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0148] Unless otherwise stated, as used herein, the term “treating” means to reverse, alleviate, or inhibit the progression of a disease or condition to which such term applies, or one or more symptoms of such a disease or condition, or to prevent such a disease or condition, or one or more symptoms of such a disease or condition.

[0149] As used herein, “individual” includes both human and non-human animals. Exemplary human individuals include human individuals suffering from a disease (such as the disease described herein) (referred to as patients) or normal individuals. In this invention, “non-human animals” includes all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock, and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.). Beneficial effects

[0150] The camptothecin derivative of this invention exhibits good activity, a more stable lactone ring, low efflux rate, and improved safety. It shows great potential as a prodrug toxin in the development of ADC drugs (e.g., drugs for treating gastrointestinal tumors). Detailed Implementation

[0151] Example

[0152] The present invention is further described below with reference to embodiments and experimental examples, but these embodiments are not intended to limit the scope of the invention.

[0153] Example 1

[0154] Synthesis route:

[0155] (1) 3-fluoro-2-methoxybenzaldehyde 1a (10 g, 64.9 mmol), 2-carboxyethyltriphenylphosphine bromide (40.4 g, 97.35 mmol), and potassium tert-butoxide (21.8 g, 194.7 mmol) were dissolved in anhydrous tetrahydrofuran (100 mL), and the reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed by TLC monitoring, the reaction mixture was diluted with ethyl acetate, washed three times with saturated brine, dried and concentrated, and separated by column chromatography (dichloromethane / methanol = 15 / 1) to obtain compound 1b (11 g).

[0156] (2) Compound 1b (11 g, 52.4 mmol) and palladium on carbon (5 g) were dissolved in anhydrous ethanol (50 mL), and the reaction mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS to ensure the reaction was complete. The reaction mixture was filtered through diatomaceous earth, and the liquid was concentrated under reduced pressure to obtain compound 1c (10 g).

[0157] (3) Compound 1c (10 g, 47.2 mmol) was dissolved in polyphosphoric acid (50 mL), and then stirred at 110 °C for 3 h. The reaction was monitored by LCMS until completion. Sodium carbonate solution was added to the reaction solution to adjust the pH to 8, and then extracted with ethyl acetate. The resulting product was purified by silica gel column chromatography to obtain compound 1d (5.2 g). LCMS (ESI) m / z: 195.2 [M+H] + .

[0158] (4) Compound 1d (5.2 g, 26.8 mmol) was dissolved in concentrated sulfuric acid (5 mL), and concentrated nitric acid (1 mL) was slowly added at 0 °C. The reaction solution was stirred at room temperature for 2 hours. The reaction of the starting materials was monitored by TLC until the reaction was complete. Ice water was added to the reaction solution to precipitate the solid. The solid was filtered off and purified by silica gel column chromatography to obtain compound 1e (2.4 g).

[0159] (5) Compound 1e (2.4 g, 10 mmol) and palladium on carbon (1 g) were dissolved in anhydrous ethanol (10 mL), and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was monitored by LCMS to indicate completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the liquid was concentrated under reduced pressure to obtain compound 1f (2 g). LCMS (ESI) m / z: 210.1 [M+H] + .

[0160] (6) Compound 1f (2.1 g, 9.57 mmol), (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f] containing indene-3,6,10(4H)-trione (2.5 g, 9.57 mmol) and p-toluenesulfonic acid (454 mg, 2.4 mmol) were dissolved in anhydrous toluene solution (10 mL) and heated to 110 °C. The mixture was stirred at this temperature for 16 hours. After the reaction was monitored by LCMS, the reaction solution was concentrated under reduced pressure and passed through a silica gel column to obtain compound 1 g (1.5 g). LCMS (ESI) m / z: 437.1 [M+H] + .

[0161] (7) 1 g (150 mg, 0.34 mmol) of compound and 2 mL of boron tribromide were dissolved in 5 mL of anhydrous dichloromethane and stirred at room temperature for 16 hours. After the reaction was completed by LCMS monitoring, the reaction solution was concentrated under reduced pressure and purified to obtain compound 1 (10 mg).

[0162] Example 2

[0163] Synthesis route:

[0164] (1) 20 g (76.3 mmol) of 1-bromo-4-fluoro-5-methyl-2-nitrobenzene 2a was dissolved in 200 mL of 1,4-dioxane. 1.9 g (3.8 mmol) of di(tri-tert-butylphosphine)palladium and 37.1 mL (228.9 mmol) of tert-butyl butyrate were added to the solution, and the mixture was stirred at 100 °C under nitrogen protection for 16 hours. The organic phase was collected by filtration, diluted with 500 mL of ethyl acetate, and washed with 500 mL of water. The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give compound 2b (18.0 g).

[0165] (2) Compound 2b (18.0 g, 55.7 mmol) was dissolved in a methanol solution (200 mL), and 10% wet palladium on carbon (1.8 g, 10% w / w) was added to the solution. The mixture was stirred at room temperature for 16 hours. The reaction solution was filtered through diatomaceous earth, the filter cake was washed with methanol, and the organic layer was concentrated to obtain crude compound 2c (17.2 g).

[0166] (3) Compound 2c (17.2 g, 52.9 mmol) was dissolved in dichloromethane (170 mL), and trifluoroacetic acid (85 mL) was added to the solution. The reaction was stirred at room temperature for 4 hours. The reaction solution was washed with saturated brine (10 mL) and extracted with dichloromethane (100 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give the target compound 2d (11.2 g).

[0167] (4) Compound 2d (11.2 g, 41.6 mmol) was dissolved in trifluoroacetic acid (110 mL), and trifluoroacetic anhydride (22 mL) was added to the solution. The reaction mixture was stirred at room temperature for 2 hours. The reaction solution was washed with saturated brine (10 mL) and extracted with ethyl acetate (100 mL). The organic layer was adjusted to alkaline pH with saturated sodium bicarbonate solution, and the organic phase was collected and concentrated. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give the target compound 2e (8.5 g).

[0168] (5) Compound 2e (500 mg, 2.0 mmol) was dissolved in anhydrous tetrahydrofuran solution (10 mL), and bis(trimethylsilylaminolithium) (6.0 mL, 1.0 M, 6.0 mmol) was slowly added dropwise to the reaction solution under nitrogen protection at -78 °C. After stirring the mixture at -78 °C for 30 minutes, a tetrahydrofuran solution (10 mL) of N-fluorobis(benzenesulfonamide) (1.88 g, 6.0 mmol) was slowly added dropwise to the above mixture. After the addition was complete, the reaction was brought to room temperature and stirred for 2 hours. The reaction solution was poured into ice water (10 mL) and extracted with ethyl acetate (10 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to obtain the target compound 2f (440 mg).

[0169] (6) 10 mL of hydrobromic acid aqueous solution was added to compound 2f (100 mg, 1.53 mmol), followed by 85 mL of trifluoroacetic acid. The reaction was refluxed and stirred at 100 °C for 6 hours. The reaction mixture was extracted with dichloromethane (20 mL x 2). The organic layer was dried over anhydrous sodium sulfate and concentrated to give 2 g (80 mg) of crude compound. LCMS (ESI) m / z: 230.1 [MH] -

[0170] (7) 2 g (80 mg, 0.35 mmol) of compound was dissolved in toluene (8 mL), and then (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f] indene-3,6,10(4H)-trione (100 mg, 0.38 mmol) and p-toluenesulfonic acid (12 mg, 0.07 mmol) were added. The mixture was sealed and heated to 130 °C with stirring for 16 hours. The reaction solution was removed by vacuum distillation, and the crude product was purified by preparative thin-layer chromatography (methanol:dichloromethane = 1:20) to obtain compound 2 (5 mg).

[0171] Example 3

[0172] Synthesis route:

[0173] (1) A mixture of 12M hydrochloric acid solution (1 mL) and 1,4-dioxane (4 mL) was added to compound 2f (100 mg, 1.53 mmol), and the resulting mixture was stirred at 60 °C for 2 hours. The reaction solution was diluted with ethyl acetate (5 mL), washed with water (5 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude compound 3a (80 mg).

[0174] (2) Compound 3a (80 mg, 0.35 mmol) was dissolved in toluene (8 mL), and then (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f] indene-3,6,10(4H)-trione (94 mg, 0.36 mmol) and p-toluenesulfonic acid (11 mg, 0.06 mmol) were added. The mixture was sealed and heated to 130 °C with stirring for 16 hours. The reaction solution was removed by vacuum distillation, and the crude product was purified by preparative thin-layer chromatography (methanol: dichloromethane = 1:20) to obtain compound 3 (45.0 mg).

[0175] Example 4

[0176] Synthesis route:

[0177] (1) Iodine (46.2 g, 182 mmol) and sodium iodate (36.0 g, 182 mmol) were added to a concentrated sulfuric acid (455 mL) solution of 1-bromo-2-fluoro-4-nitrobenzene 4a (100 g, 455 mmol) under ice-water bath conditions. The reaction mixture was slowly heated to room temperature and stirred for 48 hours. The reaction mixture was slowly poured into a saturated sodium sulfite solution at 0 °C to quench the reaction and stirred for 30 minutes. The precipitated yellow solid was filtered off, the filter cake was washed twice with water, dissolved in ethyl acetate, dried with anhydrous sodium sulfate, filtered, and concentrated to obtain the target compound 4b (80 g crude product). This crude product was used directly in the next step of the reaction without further purification.

[0178] (2) Under ice-water bath conditions, concentrated hydrochloric acid (42 mL) and stannous chloride (110 g, 578 mmol) were added to an ethanol (1000 mL) solution of compound 4b (50 g, 145 mmol). The reaction mixture was slowly heated to room temperature and stirred for 4 hours. After concentration under reduced pressure, the reaction mixture was diluted with water, adjusted to pH 10-12 with 2N sodium hydroxide solution, filtered through diatomaceous earth, and the filtrate was extracted with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, and dried with anhydrous sodium sulfate. The desiccant was removed by filtration. The diatomaceous earth filter cake was slurried through a dichloromethane:methanol (10:1) mixture and filtered. The organic solvents were combined and concentrated under reduced pressure to obtain the target compound 4c (35 g crude product). This crude product was used directly in the next reaction without further purification. LCMS (ESI) m / z: 313.7, 315.7 [MH] - .

[0179] (3) Acetic anhydride (23 mL, 244 mmol) was added to a solution of compound 4c (35 g, 111 mmol) in ethyl acetate (370 mL) under ice-water bath conditions. The reaction mixture was slowly heated to room temperature and stirred for 16 hours. The reaction solution was diluted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) and reversed-phase column chromatography (acetonitrile: 0.5% formic acid aqueous solution = 0%–40%) to obtain the target compound 4d (13 g). LCMS (ESI) m / z: 357.9, 359.9 [M+H] + .

[0180] (4) Butyric acid (0.475 mL, 5.59 mmol) and triethylamine (3.88 mL, 27.9 mmol) were added to an anhydrous N,N-dimethylformamide (30 mL) solution of compound 4d (2 g, 5.59 mmol). Under argon protection, tri-o-tolylphosphide (340 mg, 1.12 mmol) and palladium acetate (125 mg, 0.559 mmol) were added to the reaction solution. Six parallel reactions were simultaneously heated and stirred at 80 °C for 3 hours. The reaction solutions were combined, diluted with water, adjusted to pH 3-4 with 2N hydrochloric acid solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The desiccant was removed by filtration and the solution was concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (methanol:dichloromethane = 0%–10%) to obtain the target compound 4e (1.5 g). LCMS(ESI) m / z: 315.9, 317.9 [M+H] + .

[0181] (5) Platinum dioxide (150 mg) was added to a tetrahydrofuran (50 mL) solution of compound 4e (1.5 g, 5.59 mmol). The reaction was stirred at room temperature under hydrogen pressure for 4 hours. The reaction solution was filtered through diatomaceous earth to obtain platinum dioxide. The filter cake was washed twice with methanol, and the filtrates were combined and concentrated under reduced pressure to obtain the target compound 4f (1.4 g crude product). This crude product was used directly in the next step without further purification. LCMS (ESI) m / z: 316.0, 318.0 [MH] - .

[0182] (6) Trifluoroacetic anhydride (6.12 mL, 44.0 mmol) was added to a solution of compound 4f (1.4 g, 4.40 mmol) in 6 mL of trifluoroacetic acid. The reaction was stirred at 30 °C for 24 hours. The reaction mixture was diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (ethyl acetate: petroleum ether = 0%–15%) to give 4 g (500 mg) of the target compound. LCMS (ESI) m / z: 300.0, 302.0 [M+H] + .

[0183] (7) At -65°C, 1M bis(trimethylsilylamino)lithium (3.17 mL, 3.17 mmol) was slowly added dropwise to an argon-protected solution of 4 g (475 mg, 1.58 mmol) of the compound in 3 mL of anhydrous tetrahydrofuran. After stirring the reaction solution at -65°C for 0.5 hours, a solution of N-fluorobis(benzenesulfonamide) (749 mg, 2.37 mmol) in 1 mL of tetrahydrofuran was slowly added dropwise to the above reaction solution, and the mixture was brought to room temperature and stirred for 2 hours. The reaction solution was quenched with saturated ammonium chloride aqueous solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated sodium chloride solution and dried with anhydrous sodium sulfate. The desiccant was removed by filtration and the solution was concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (ethyl acetate: petroleum ether = 0%–15%) to obtain the target compound 4 h (192 mg). LCMS (ESI) m / z: 336.0, 338.0.

[0184] (8) 3 mL of 4 M hydrochloric acid-methanol solution was added to compound 4h (100 mg, 0.298 mmol). The reaction mixture was stirred at room temperature for 16 hours. The solvent was removed by concentration under reduced pressure to obtain the target compound 4i (80 mg). LCMS (ESI) m / z: 294.0, 296.0 [M+H] + .

[0185] (9) In a 5 mL microwave tube, potassium N-aminomethyltrifluoroborate (42 mg, 0.179 mmol), cesium carbonate (166 mg, 0.510 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (14 mg, 0.017 mmol) were added to a solution of compound 4i (50 mg, 0.170 mmol) in 1 mL of 1,4-dioxane and water (0.2 mL). The reaction mixture was sealed under argon protection and heated at 110 °C for 16 hours. The reaction mixture was diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and concentrated under reduced pressure. The crude product was purified by preparative agar (petroleum ether:ethyl acetate = 4:1) to obtain the target compound 4j (14 mg). LCMS (ESI) m / z: 343.4 [MH] - .

[0186] (10) In a 5 mL microwave tube, (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f] indene-3,6,10(4H)-trione (11 mg, 0.407 mmol) and p-toluenesulfonic acid (1.4 mg, 0.008 mmol) were added to a mixture of compound 4j (14 mg, 0.407 mmol) and toluene (5 mL). The reaction mixture was sealed and heated at 130 °C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent, and the crude product was purified by preparative chromatography (dichloromethane:methanol = 10:1) and high performance liquid chromatography (acetonitrile:formic acid aqueous solution) to obtain target compound 4 (2 mg).

[0187] Example 5

[0188] Synthesis route:

[0189] Formaldehyde (0.764 mg, 0.025 mmol) and formic acid (5 mg, 0.106 mmol) were added to a methanol (1 mL) solution of compound 4 (10 mg, 0.021 mmol). The reaction mixture was stirred at room temperature for 1 hour under argon protection. Sodium cyanoborohydride (2 mg, 0.032 mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for another 16 hours under argon protection. The reaction mixture was purified by preparative thin-layer chromatography (dichloromethane:methanol = 10:1) to give compound 5 (6 mg).

[0190] Example 6

[0191] Synthesis route:

[0192] (1) 100 g (526.27 mmol) of 3-bromo-5-fluoroaniline 6a was dissolved in 1 L of anhydrous N,N-dimethylformamide solution. 77.30 g (578.90 mmol) of NCS was added to the mixture, and the mixture was stirred at 60 °C for 6 hours. After the reaction was completed as monitored by LCMS, the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain the target compound 6b (47 g). LCMS (ESI) m / z: 226.1 [M+H] + .

[0193] (2) Compound 6b (47 g, 209.39 mmol) was dissolved in anhydrous ethyl acetate (500 mL). Acetic anhydride (32.07 g, 314.09 mmol) was added to the mixture, and the mixture was stirred at 50 °C for 16 hours. After the reaction was completed as monitored by LCMS, the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain the target compound 6c (39 g). LCMS (ESI) m / z: 267.9 [M+H] + .

[0194] (3) Compound 6c (39 g, 146.34 mmol) was dissolved in anhydrous tetrahydrofuran solution (400 mL), and n-butyllithium (122.93 mL, 2.5 M, 307.32 mmol) was slowly added dropwise to the reaction solution under nitrogen protection at -65 °C. After stirring the mixture at -65 °C for 1 hour, a tetrahydrofuran solution (100 mL) of tert-butyl 3-oxocyclobutane-1-carboxylate (37.36 g, 219.52 mmol) was slowly added dropwise to the above mixture, and the resulting mixture was stirred at -65 °C for 0.5 hours. The reaction was slowly raised to room temperature and stirred for another 16 hours. After the reaction was completed by LCMS monitoring, the reaction solution was poured into a saturated ammonium chloride solution (200 mL) and extracted with ethyl acetate (200 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The concentrated crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give the target compound 6d (7.3 g). LCMS (ESI) m / z: 356.2 [MH] - .

[0195] (4) Compound 6d (4 g, 11.18 mmol) was dissolved in anhydrous trifluoroacetic acid solution (40 mL). Triethylsilane (40 mL) was added to the mixture, and the mixture was stirred at 50 °C for 16 hours. After the reaction was monitored by LCMS, the reaction solution was poured into water (100 mL), the pH was adjusted to 6 with saturated sodium bicarbonate, and the mixture was extracted with dichloromethane (1 L). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to high performance liquid chromatography (acetonitrile, trifluoroacetic acid aqueous solution system) to obtain the target compound 6e (1.8 g). LCMS (ESI) m / z: 286.0 [M+H] + .

[0196] (5) Compound 6e (0.5 g, 1.75 mmol) was dissolved in anhydrous trifluoroacetic acid solution (5 mL). Trifluoroacetic anhydride (5 mL) was added to the mixture, and the mixture was stirred at 100 °C for 8 hours. After the reaction was monitored by LCMS, the reaction solution was poured into water (100 mL), the pH was adjusted to 7 with saturated sodium bicarbonate, and extracted with ethyl acetate (50 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain the target compound 6f (0.117 g). LCMS (ESI) m / z: 268.1 [M+H] + .

[0197] (6) Compound 6f (0.117 g, 0.437 mmol) was dissolved in dioxane (4 mL). Palladium acetate (9.81 mg, 0.043 mmol), S-(-)-1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (54.43 mg, 0.087 mmol), diphenylmethaneimine (118.82 mg, 0.655 mmol, 1.5 equivalents), and cesium carbonate (427.23 mg, 1.31 mmol, 3 equivalents) were added to the mixture, and the mixture was stirred at 100 °C for 16 hours. After the reaction was completed as monitored by LCMS, the solvent was removed by vacuum distillation. The crude product was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain the target compound 6 g (0.051 g). LCMS (ESI) m / z: 413.1 [M+H] + .

[0198] (7) 6 g (0.051 g, 0.123 mmol) of compound was dissolved in 4 M HCl / EA solution (4 mL). The mixture was stirred at room temperature for 2 hours. After the reaction was completed by LCMS monitoring, the solvent was removed by vacuum distillation. The crude product obtained by concentration was slurryed with diethyl ether to obtain the target compound 6 h (0.025 g). LCMS (ESI) m / z: 207.2 [M+H] + .

[0199] (8) Compound 6h (0.025 g, 0.121 mmol) was dissolved in anhydrous toluene solution (4 mL). p-Toluenesulfonic acid (4.18 mg, 0.024 mmol) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f]-indene-3,6,10(4H)-trione (47.87 mg, 0.18 mmol, 1.5 equivalents) were added to the mixture, and the resulting mixture was stirred at 130 °C for 16 hours. After the reaction was monitored by LCMS, the solvent was removed by vacuum distillation. The crude product obtained by concentration was used to prepare the target compound 6 (1.1 mg) by high performance liquid chromatography (acetonitrile, trifluoroacetic acid aqueous solution system).

[0200] Example 7

[0201] Synthesis route:

[0202] (1) Compound 7a (10 g, 38.16 mmol) was dissolved in anhydrous tetrahydrofuran solution (100 mL), and n-butyllithium (32.05 mL, 2.5 M, 80.13 mmol) was slowly added dropwise to the reaction solution under nitrogen protection at -65 °C. After stirring the mixture at -65 °C for 1 hour, a tetrahydrofuran solution (50 mL) of tert-butyl 3-oxocyclobutane-1-carboxylate (9.74 g, 57.23 mmol) was slowly added dropwise to the above mixture, and the resulting mixture was stirred at -65 °C for 0.5 hours. The reaction was slowly raised to room temperature and stirred for another 16 hours. After the reaction was completed by LCMS monitoring, the reaction solution was poured into a saturated ammonium chloride solution (100 mL) and extracted with ethyl acetate (200 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The concentrated crude product was subjected to high-performance liquid chromatography (HPLC) (acetonitrile, trifluoroacetic acid aqueous solution system) to obtain the target compound 7b (3.9 g). LCMS (ESI) m / z: 352.2 [M+H] - .

[0203] (2) Compound 7b (3.9 g, 11.04 mmol, 1.00 equivalent) was dissolved in anhydrous trifluoroacetic acid solution (30 mL). Triethylsilane (30 mL) was added to the mixture, and the mixture was stirred at 50 °C for 16 hours. After the reaction was monitored by LCMS, the reaction solution was poured into water (100 mL), the pH was adjusted to 6 with saturated sodium bicarbonate, and the solution was extracted with dichloromethane (1 L). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product obtained by concentration was subjected to high performance liquid chromatography (acetonitrile, trifluoroacetic acid aqueous solution system) to obtain the target compound 7c (1.3 g). LCMS (ESI) m / z: 282.2 [M+H] + .

[0204] (3) Compound 7c (1.3 g, 4.62 mmol) was dissolved in anhydrous trifluoroacetic acid solution (15 mL), and trifluoroacetic anhydride (15 mL) was added to the mixture. The resulting mixture was stirred at 100 °C for 8 hours. After the reaction was monitored by LCMS, the reaction solution was poured into water (300 mL), the pH was adjusted to 7 with saturated sodium bicarbonate, and the mixture was extracted with ethyl acetate (200 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain the target compound 7d (0.31 g). LCMS (ESI) m / z: 264.1 [M+H] + .

[0205] (4) Compound 7d (0.31 g, 1.18 mmol) was dissolved in 4 M hydrochloric acid / methanol (10 mL), and the mixture was stirred at room temperature for 16 hours. After the reaction was completed as monitored by LCMS, the solvent was removed by vacuum distillation. The crude product was concentrated to obtain the target compound 7e (0.26 g). LCMS (ESI) m / z: 222.1 [M+H] + .

[0206] (5) Compound 7e (0.26 g, 1.18 mmol) was dissolved in anhydrous toluene solution (15 mL). p-Toluenesulfonic acid (40.47 mg, 0.235 mmol) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f] indene-3,6,10(4H)-trione (464.07 mg, 1.76 mmol) were added to the mixture, and the mixture was stirred at 130 °C for 16 hours. After the reaction was monitored by LCMS, the solvent was removed by vacuum distillation. The concentrated crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give the target compound 7f (0.083 g, 0.185 mmol, yield 15.75%) as a brown solid. LCMS (ESI) m / z: 449.1 [M+H]+ .

[0207] (6) Compound 7f (83 mg, 0.185 mmol, 1 equivalent) was dissolved in dichloromethane (4 mL), and boron tribromide (238.14 mg, 0.925 mmol) was added to the reaction solution. The mixture was stirred at 0 °C for 2 hours. After the reaction was completed by LCMS monitoring, the mixture was quenched with methanol at 0 °C, and the reaction solution was concentrated under reduced pressure. 7 (11.24 mg) was obtained by preparative purification.

[0208] Example 8

[0209] Synthesis route:

[0210] In a 20 mL sealed tube, (R)-5-ethyl-5-hydroxy-4,5,8,9-tetrahydro-1H,3H-oxabino[3,4-f] indene-3,7,11-trione (113 mg, 0.406 mmol), p-toluenesulfonic acid (38 mg, 0.222 mmol, 0.6 equivalents), and anhydrous magnesium sulfate (1 g) were added to a mixture of compound 8a (100 mg, 0.369 mmol) and acetic acid (10 mL). The reaction mixture was sealed under argon protection and heated at 105 °C for 12 hours. After cooling, the mixture was diluted with ethyl acetate, filtered, and the filter cake was washed with ethyl acetate. The filtrate was concentrated under reduced pressure to remove the solvent. The crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol = 10:1) and preparative high-performance liquid chromatography (0.5% formic acid aqueous solution, acetonitrile) to obtain the target compounds 8-1 (0.94 mg) and 8-2 (1.95 mg).

[0211] Example 9

[0212] Synthesis route:

[0213] (1) Benzo[d][1,3]dioxane-5-amine 9a (20 g, 146 mmol) was dissolved in a mixture of dichloromethane (500 mL) and methanol (250 mL). Tetrabutylammonium tribromide (80.2 g, 166 mmol) was added to the solution, and the resulting mixture was stirred at room temperature for 20 minutes. The solution was diluted with dichloromethane (1000 mL) and washed with 10% sodium thiosulfate solution (2 x 1000 mL) and saturated brine (1000 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give the target compound 9b (15 g).

[0214] (2) Compound 9b (9.5 g, 44.0 mmol) was dissolved in ethyl acetate (100 mL), and acetic anhydride (9 mL, 96.7 mmol) was added to the solution. The resulting mixture was stirred overnight at room temperature. The solution was diluted with ethyl acetate (500 mL) and washed with saturated brine (500 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to give the target compound 9c (10.7 g).

[0215] (3) Compound 9c (5 g, 19.4 mmol) was dissolved in anhydrous tetrahydrofuran solution (50 mL), and n-butyllithium (17 mL, 2.5 M, 43.6 mmol, 2.25 equivalents) was slowly added dropwise to the reaction solution under nitrogen protection at -65 °C. After stirring the mixture at -65 °C for 1 hour, a tetrahydrofuran solution (20 mL) of 3-(benzyloxy)-N-methoxy-N-methylcyclobutane-1-carboxamide (4.35 g, 17.4 mmol) was slowly added dropwise to the above mixture, and the resulting mixture was stirred at -65 °C for 1 hour. The reaction was quenched by adding saturated ammonium chloride solution (100 mL), and the resulting mixture was extracted with ethyl acetate (100 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to give the target compound 9d (2.68 g). LCMS(ESI) m / z: 368.2 [M+H] + .

[0216] (4) 7 mL of 4 M hydrochloric acid-methanol solution was added to compound 9d (250 mg, 0.680 mmol), and the resulting mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure to obtain the target compound 9e (200 mg). LCMS (ESI) m / z: 326.1 [M+H] +

[0217] (5) Compound 9e (150 mg, 0.461 mmol) was dissolved in a mixed solution of glacial acetic acid (1 drop), methanol (1 mL), and tetrahydrofuran (5 mL). Palladium on carbon (30 mg) and palladium hydroxide (30 mg) were then added. The mixture was heated to 65 °C under hydrogen pressure and stirred for 4 hours. After cooling to room temperature, the reaction solution was filtered through diatomaceous earth. The filter cake was washed with methanol to obtain the filtrate, which was then distilled under reduced pressure to obtain the target compound 9f (100 mg). LCMS (ESI) m / z: 236.1 [M+H] + .

[0218] (6) 9f (100 mg, 0.425 mmol) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f] containing indene-3,6,10(4H)-trione (112 mg, 0.425 mmol) and p-toluenesulfonic acid (15 mg, 0.085 mmol) were weighed into a microwave tube, toluene (2 mL) was added, the tube was sealed, and the mixture was stirred at 130 °C for 16 hours. The reaction solution was removed by vacuum distillation. The crude product was dissolved in N,N-dimethylformamide and filtered. The solution was prepared by high performance liquid chromatography (acetonitrile, trifluoroacetic acid aqueous solution system), and lyophilized to obtain target compound 9 (10.6 mg).

[0219] Example 10

[0220] Synthesis route:

[0221] (1) 1-Bromo-4-fluoro-5-methyl-2-nitrobenzene 10a (2 g, 8.55 mmol, 1.00 equivalent) was dissolved in tetrahydrofuran (20 mL). Triphenylphosphine palladium dichloride (899 mg, 12.8 mmol), cuprous iodide (325 mg, 1.28 mmol), triethylamine (3.56 mL, 25.64 mmol), and but-3-yn-1-ol (0.97 mL, 12.8 mmol) were added to the solution, and the resulting mixture was stirred at 70 °C under nitrogen protection for 16 hours. The solution was filtered, the organic phase was collected, diluted with ethyl acetate (50 mL), and washed with water (50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give the target compound 10b (1.1 g).

[0222] (2) Compound 10b (1.1 g, 4.9 mmol) was dissolved in a mixture of ethanol and water (20 mL). Tin (1.17 g, 9.96 mmol), sodium sulfide (115 mg, 1.48 mmol), and concentrated hydrochloric acid (4.18 mL, 49.3 mmol) were added to the solution, and the mixture was stirred at 78 °C for 2 hours. The solution was adjusted to alkalinity with saturated sodium sulfite and washed with ethyl acetate (10 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give the target compound 10c (630 mg).

[0223] (3) Compound 10c (630 mg, 3.26 mmol, 1.00 equivalent), acetyl chloride (0.58 mL, 8.15 mmol, 2.5 equivalent), and triethylamine (2.27 mL, 16.3 mmol, 5.00 equivalent) were dissolved in dichloromethane (6 mL) at 0 °C and stirred at room temperature for 16 hours. The reaction mixture was washed with saturated brine (10 mL) and extracted with dichloromethane (10 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to give the target compound 10d (390 mg, 1.32 mmol, yield 47%) as a white solid.

[0224] (4) Compound 10d (390 mg, 1.32 mmol) was dissolved in anhydrous tetrahydrofuran solution (4 mL), and bis(trimethylsilylaminolithium) (6.6 mL, 1.0 M, 6.6 mmol) was slowly added dropwise to the reaction solution under nitrogen protection at -65 °C. After stirring the mixture at -65 °C for 30 min, a tetrahydrofuran solution of N-fluorobis(benzenesulfonamide) (1.67 g, 5.28 mmol) (15 mL) was slowly added dropwise to the above mixture, and the resulting mixture was stirred at -65 °C for 1 h. The reaction was slowly raised to room temperature and stirred for another 7 h. The reaction solution was poured into ice water (10 mL) and extracted with ethyl acetate (10 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to give the target compound 10e (57 mg).

[0225] (5) A mixture of 12M hydrochloric acid solution (0.3 mL) and 1,4-dioxane (0.6 mL) was added to 10e (57 mg, 0.2 mmol), and the resulting mixture was stirred at 60 °C for 2 hours. The reaction solution was diluted with ethyl acetate (5 mL), washed with water (5 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 10f (57 mg). LCMS (ESI) m / z: 230.1 [M-OH] + .

[0226] (6) 10f (57 mg, 0.23 mmol) was dissolved in toluene (1 mL), and then (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f] containing indene-3,6,10(4H)-trione (73 mg, 0.280 mmol) and p-toluenesulfonic acid (8 mg, 0.05 mmol) were added. The mixture was sealed and heated to 130 °C with stirring for 16 hours. The reaction solution was removed by vacuum distillation. The crude product was dissolved in N,N-dimethylformamide and filtered. It was prepared by high performance liquid chromatography (acetonitrile, trifluoroacetic acid aqueous solution system) and lyophilized to obtain target compound 10 (6.9 mg).

[0227] Example 11

[0228] Synthesis route:

[0229] (1) 5-Bromobenzo[d][1,3]dioxane 11a (68 g, 338.3 mmol) was dissolved in a mixture of acetic acid (410 mL) and water (3 mL) at room temperature. Fuming nitric acid (136 mL) was added to the mixture at 0 °C, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was poured into ice water (500 mL) to precipitate a solid. The solid was filtered and washed with water until the pH of the solution was neutral. The filter cake was collected, dried, and the target compound 11b (77.0 g) was obtained.

[0230] (2) Compound 11b (5.0 g, 20.3 mmol) was dissolved in N,N-dimethylformamide solution (25 mL). Buty-3-yn-1-ol (3.1 mL, 40.6 mmol), bis(triphenylphosphine)palladium dichloride (428 mg, 0.6 mmol), cuprous iodide (193 mg, 1.02 mmol), and triethylamine (7.1 mL, 50.8 mmol) were added to the solution. The reaction mixture was stirred at 50 °C for 3 hours. The reaction mixture was poured into 1% ammonia water (50 mL), stirred for 30 minutes, filtered, and the filter cake was collected. Water (50 mL) was added to the filter cake, and the mixture was stirred for 20 minutes. The mixture was then filtered. After drying the filter cake, compound 11c (4.1 g) was obtained.

[0231] (3) Compound 11c (1.0 g, 4.25 mmol), tin (1.0 g, 8.5 mmol), sodium sulfide (99 mg, 1.28 mmol), and concentrated hydrochloric acid (3.5 mL, 42.5 mmol) were added to a mixed solution of ethanol (18 mL) and water (2 mL) at room temperature. The mixture was heated to 78 °C and stirred for 2 hours. After the reaction was complete, the mixture was filtered while hot and concentrated. The crude product was pulped with ethyl acetate, and the filter cake was collected. The filter cake was adjusted to alkalinity with sodium bicarbonate and extracted with ethyl acetate. The organic phase was dried and concentrated to obtain compound 11d (512 mg).

[0232] (4) Compound 11d (51 mg, 0.23 mmol) was dissolved in toluene (1 mL), and then (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f] indene-3,6,10(4H)-trione (73 mg, 0.280 mmol) and p-toluenesulfonic acid (8 mg, 0.05 mmol) were added. The mixture was sealed and heated to 130 °C with stirring for 16 hours. The reaction solution was removed by vacuum distillation. The crude product was dissolved in N,N-dimethylformamide and filtered. It was prepared by high performance liquid chromatography (acetonitrile, trifluoroacetic acid aqueous solution system) and lyophilized to obtain target compound 11 (6.1 mg).

[0233] Example 12

[0234] Synthesis route:

[0235] Compound 12a (50 mg, 0.22 mmol), p-toluenesulfonic acid (7.6 mg, 0.044 mmol), and INT1-6 (61.5 mg, 0.22 mmol) were dissolved in toluene (5 mL) and reacted at 120 °C for 16 hours. After the reaction was completed as monitored by LCMS, the reaction solution was cooled to room temperature, filtered, and the filter cake was prepared by pre-HPLC (acetonitrile:water (formic acid)) to give compound 12 (13 mg).

[0236] Example 13

[0237] Synthesis route:

[0238] (1) Acetyl chloride (164 mg, 3.1 mmol) and triethylamine (423 mg, 4.2 mmol) were dissolved in dichloromethane (6 mL) containing 1-(2-amino-4-fluoro-5-methoxyphenyl)-4-hydroxybut-1-one 15a (190 mg, 0.84 mmol) at 0 °C and stirred at room temperature for 16 hours. The reaction mixture was washed with saturated brine (10 mL) and extracted with dichloromethane (10 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. The crude product was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to give the target compound 4-(2-acetamido-4-fluoro-5-methoxyphenyl)-4-oxobutylacetate 15b (140 mg). LCMS (ESI) m / z: 312.1 [M+H] + .

[0239] (2) NFSI (567 mg, 1.8 mmol), 1,3-dimethylimidazolinone (0.2 mL, 1.8 mmol), and compound 15b (140 mg, 0.45 mmol) were dissolved in anhydrous tetrahydrofuran solution (5 mL). LiHMDS (2.25 mL, 1 M, 2.25 mmol) was slowly added dropwise to the reaction solution under nitrogen protection at -65 °C. The addition was completed in approximately 1 hour, and the reaction was continued to be stirred at -65 °C for 3 hours. The reaction was quenched by adding saturated sodium thiosulfate solution (10 mL), the temperature was raised to room temperature, and stirring was continued for 1 hour. The mixture was then extracted with ethyl acetate (5 mL x 3). The combined organic phases were washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reversed-phase rapid column chromatography (0.1% formic acid aqueous solution, acetonitrile). The eluent was neutralized with saturated sodium bicarbonate and extracted with ethyl acetate (10 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the target compound 15c (40 mg, 0.13 mmol).

[0240] (3) A mixture of 12M hydrochloric acid solution (0.3 mL) and 1,4-dioxane (0.6 mL) was added to N-(2-(2,2-difluoro-4-hydroxybutyryl)-5-fluoro-4-methoxyphenyl)acetamide 15c (40 mg, 0.13 mmol), and the resulting mixture was stirred at 60 °C for 2 hours. The reaction solution was diluted with ethyl acetate (5 mL) and washed with water (5 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude 1-(2-amino-4-fluoro-5-methoxyphenyl)-2,2-difluoro-4-hydroxybut-1-one 15d (43 mg).

[0241] (4) Dissolve 1-(2-amino-4-fluoro-5-methoxyphenyl)-2,2-difluoro-4-hydroxybut-1-one 15d (43mg, 0.16mmol) in toluene (1mL), then add (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f] indene-3,6,10(4H)-trione (52mg, 0.2mmol) and p-toluenesulfonic acid (6mg, 0.03mmol), seal the mixture and heat it to 130°C and stir for 16 hours. The reaction solution was removed by vacuum distillation. The crude product was dissolved in N,N-dimethylformamide and filtered. It was prepared by high performance liquid chromatography (acetonitrile and trifluoroacetic acid aqueous solution system). The product was lyophilized to obtain the target compound 15,(S)-11-(1,1-difluoro-3-hydroxypropyl)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-1,12-dihydro-14H-pyrano[3',4':6,7] indene[1,2-b]quinoline-3,14(4H)-dione (12 mg).

[0242] Example 14

[0243] Step 1: Synthesize compound 26-1

[0244] Compound 15 (200 mg, 0.407 mmol, 1 equivalent) was dissolved in dichloromethane (4 mL). Carbon tetrabromide (534 mg, 2.04 mmol, 5 equivalents) and triphenylphosphine (270 mg, 0.815 mmol, 2 equivalents) were added under ice bath conditions. The reaction mixture was stirred at room temperature for 3 hours. The reaction solution was quenched with saturated sodium chloride aqueous solution (10 mL) and extracted with dichloromethane (10 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. The crude product was subjected to rapid silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give the target compound 26-1 (100 mg).

[0245] Step 2: Synthesize compound 26

[0246] (3-fluorobicyclo[1.1.1]pent-1-yl)methylamine hydrochloride (14.6 mg, 0.144 mmol, 2 equivalents) was dissolved in N,N-dimethylformamide (2 mL), and triethylamine (22 mg, 0.216 mmol, 3 equivalents) was added. The mixture was stirred at room temperature for 10 minutes, and then compound 26-1 (40 mg, 0.072 mmol, 1 equivalent) and sodium iodide (16 mg, 0.108 mmol, 1.5 equivalents) were added. The reaction mixture was stirred at 50 °C for 16 hours. The reaction solution was prepared by preparative high performance liquid chromatography (acetonitrile, formic acid aqueous solution system), and lyophilized to obtain target compound 26 (5 mg). 1 H NMR (400MHz, DMSO-d6) δ8.19–8.13(m,1H),7.56(d,J=8.8Hz,1H),7.34(s,1H),6.56(s,1H),5.44(s ,2H),5.38(s,2H),4.11(s,3H),3.09(s,2H),2.90(s,2H),2.06–1.83(m,8H),0.87(t,J=7.3Hz,3H).

[0247] Other compounds disclosed herein can be prepared by methods similar to those described in the above embodiments (with appropriate modifications if necessary).

[0248] Experimental Example 1: Cell Viability Test

[0249] The inhibitory activity of the drug compounds on the in vitro proliferation of SNU-5, MKN-45, SW480, LS1034, and AsPC-1 tumor cells was tested. After 6 days of culture, cell proliferation was detected using CGT reagent (CellTiter-Glo). The inhibitory activity was determined based on the IC50 value. 50 The in vitro activity of the compound was determined by measuring its value.

[0250] Note: DXd's sensitivity to killing tumor cells: SNU-5 (high), MKN-45 (medium), SW480 (medium), LS1034 (low) and AsPC-1 (low).

[0251] A. Materials and Equipment

[0252] Reagents and consumables: SNU-5 (ATCC), MKN-45 (CTCC), SW480 (ATCC), LS1034 (ATCC), and AsPC-1 (ATCC); IMDM medium (Gibco), RPMI-1640 medium (Gibco), Leibovitz's L-15 medium (Gibco), trypsin (Gibco), fetal bovine serum (Vivacell), penicillin and streptomycin (Invitrogen), phenol blue (Biological Industries), DPBS (Biosera), DMSO (Sigma), Staurosporine 96 (Selleck), CellTiter-Glo (Promega), 96-well cell culture plates (Corning), 96-well V-plates (Corning)

[0253] Instruments: Inverted microscope (OLYMPUS), CO2 incubator (ThermoFisher), centrifuge (Eppendorf), cell counter (Shanghai Meng Microbial Medical), multi-functional microplate reader (PerkinElmer).

[0254] B. Testing Methods

[0255] Experimental steps:

[0256] 1. Culture SNU-5, MKN-45, SW480, LS1034 and AsPC-1 in their respective cell cultures (see Table 1) until the confluence is about 70-90%.

[0257] 2. Passage the cells (cells should be passaged 2-15 after revival) and resuspend the cells in the corresponding complete culture medium (see Table 1). After cell counting (it is recommended that the cell viability be ≥90%), record the cell passage number and viability.

[0258] 3. Seed 90 μL of cells into each well of a 96-well plate and incubate overnight at 37°C.

[0259] 4. The following day, prepare the drug according to the preparation protocol in Table 2. Take out the 96-well plate and add 10 μL of 10x test drug, positive control staurosporine, or carrier control to each well. The total system volume is 100 μL.

[0260] 5. After mixing, place the cell culture plate in a 37°C, 5% CO2 or 37°C, 0% CO2 (SW480 cells) incubator and continue incubation for 6 days.

[0261] 6. After incubation, remove the cell culture plate and allow it to equilibrate to room temperature.

[0262] 7. Add 50 μL of CellTiter Glo reagent to each well, lyse at room temperature for 2 minutes, and then equilibrate for 10 minutes.

[0263] 8. Use an Envision microplate reader to read the plate and detect the luminescence value.

[0264] 9. Data Analysis: Using the four-parameter fitting formula in GraphPad software, an S-shaped curve of cell survival was plotted and graphed to calculate the IC50 of drug-induced cell killing.

[0265] Table 1. Cell lines

[0266] Table 2. Medication Dispensing Plan

[0267] The in vitro inhibitory activity against tumor cell proliferation was tested according to the above experimental steps, and the results are shown in Table 3.

[0268] Table 3

[0269] *DXd's cytotoxic IC50 value in various cell lines 50 The value is the average of multiple measurements.

[0270] Experimental results showed that compounds 1, 3, and 11 exhibited significantly better cytotoxic activity than Dxd on all five tumor cell lines.

[0271] Experimental Example 2: Assay of the extracellular excretion rate of camptothecin derivative Caco-2

[0272] A. Cell culture and seeding plates

[0273] Cell culture was performed using high-glucose DMEM medium containing L-glutamine, supplemented with 10% fetal bovine serum, 0.1 mg / mL streptomycin, 100 units of penicillin, and 1× non-essential amino acids.

[0274] Caco-2 cells were cultured in cell culture flasks. The incubator was set to 37°C, 5% CO2, and 95% relative humidity. Cells were ready for Transwell inoculation when confluence reached 70-90%.

[0275] Before cell seeding, add 50 μL of cell culture medium to each well in the upper chamber of the Transwell and 25 mL of cell culture medium to the lower culture plate. After incubating the culture plate in a 37°C, 5% CO2 incubator for 1 hour, it can be used for cell seeding.

[0276] After cell digestion, the cell suspension was transferred to a round-bottom centrifuge tube and centrifuged at 120g for 5 minutes.

[0277] The cells were resuspended in culture medium to a final concentration of 6.86 × 10⁻⁶. 5 Cells / mL. Add 50 μL of cell suspension to each well of a 96-well Transwell plate, resulting in a final seeding density of 2.4 × 10⁶ cells / mL. 5 cells / cm 2 .

[0278] Change the medium 48 hours after inoculation and culture for 14-18 days, changing the medium every other day.

[0279] The process of changing the culture medium is as follows: separate the Transwell chamber from the receiving plate, discard the culture medium in the receiving plate first, then discard the culture medium in the Transwell chamber, and finally add 75 μL of fresh culture medium to each chamber and 25 mL of fresh culture medium to the receiving plate.

[0280] B. Evaluation of cell monolayer membrane integrity

[0281] After 14-18 days of culture, Caco-2 should have completely merged and differentiated. At this point, it can be used for the breakthrough test.

[0282] The resistance of the single-layer film was measured using a resistance meter (Millipore, USA), and the resistance of each pore was recorded.

[0283] After the assay was completed, the Transwell culture plate was returned to the incubator.

[0284] Calculation of resistance value: Measured resistance value (ohms) × film area (cm²) 2 ) = TEER value (ohm·cm) 2 ),

[0285] If the TEER value is <230 ohms·cm 2 If so, the hole cannot be used for a penetration test.

[0286] C. Solution preparation

[0287] To prepare 1L of HBSS (10mM HEPES, pH 7.4): Weigh 2.38g HEPES and 0.35g sodium bicarbonate, add 900mL of pure water to dissolve them, then add 100mL of 10×HBSS and stir well. Adjust the pH to 7.4 and finally filter.

[0288] Prepare a high-concentration DMSO stock solution of the test substance and dilute it to 1 mM with DMSO. Then, dilute it accordingly with HBSS (10 mM HEPES, pH 7.4) to obtain a test concentration of 5 μM.

[0289] High-concentration DMSO stock solutions of the control drugs digoxin and metoprolol were prepared and diluted to 2 mM with DMSO. Subsequently, they were diluted accordingly with HBSS (10 mM HEPES, pH 7.4) to obtain a test concentration of 10 μM.

[0290] D. Drug penetration test

[0291] Remove the Transwell culture plate from the incubator. Rinse the cell monolayer twice with HBSS (10mM HEPES, pH 7.4) buffer and incubate at 37°C for 30 minutes.

[0292] The transport rate of the compound from the top to the base was determined. 125 μL of the dosing end solution was added to each well of the upper chamber (top), followed by a 50 μL sample transferred to 200 μL of acetonitrile containing an internal standard. This was used as the 0-minute dosing sample from the top of the Transwell plate for analysis. 235 μL of the receiving end solution was added to each well of the lower chamber (base).

[0293] The transport rate of the compound from the base to the top was determined. 75 μL of receiver solution was added to each well in the upper chamber (top), and 285 μL of dosing solution was added to each well in the lower chamber (base). Then, 50 μL of the sample was transferred to 200 μL of acetonitrile containing an internal standard as the 0-minute dosing sample from the base of the Transwell plate for analysis.

[0294] After merging the upper and lower transfer devices, incubate at 37°C for 2 hours.

[0295] Transfer 50 μL of sample from the working solution preparation plate to 200 μL of acetonitrile containing the internal standard as a 0-minute dosing sample for testing.

[0296] After incubation, 50 μL of sample was taken from each well of the upper and lower chambers of the Transwell plate and added to a new sample tube. 200 μL of acetonitrile containing the internal standard was added to the sample tube, vortexed for 10 minutes, and then centrifuged at 3220 g for 30 minutes. 150 μL of the supernatant was collected, diluted with an equal volume of water, and then analyzed by LC-MS / MS. All samples were prepared in duplicate.

[0297] The integrity of the cell monolayer was assessed after 2 hours of incubation using fluorescein (LY) leakage. Fluorescein stock solution was diluted to a final concentration of 100 μM using HBSS (10 mM HEPES, pH 7.4). 100 μL of fluorescein solution was added to each well of the upper Transwell plate, and 300 μL of HBSS (10 mM HEPES, pH 7.4) was added to each well of the lower receiving plate. After incubation at 37°C for 30 minutes, 80 μL of solution was aspirated from both the upper and lower layers of each well into a new 96-well plate. Fluorescence was measured using a microplate reader at excitation wavelength of 485 nm and emission wavelength of 530 nm.

[0298] E. Data Analysis

[0299] All calculations were performed using Microsoft Excel. The apparent permeability coefficient (P0.05) of the compound in Caco-2 cells was calculated based on the specific concentrations at the receiving and administering ends. app (Unit: cm / s), calculated using the following formula:

[0300] in:

[0301] V A This refers to the volume of the solution at the receiving end (0.235 mL from the top to the base, and 0.075 mL from the base to the top).

[0302] Area is the area of ​​the Transwell-96-well plate membrane (0.143 cm²). 2 );

[0303] time is the incubation time (unit: seconds);

[0304] [drug] receiver The drug concentration at the receiving end;

[0305] [drug]initial,donor is the initial drug concentration at the dosing end.

[0306] The efflux ratio is calculated using the following formula:

[0307] in:

[0308] Papp(B-A) The apparent permeability coefficient is measured from the base to the tip.

[0309] P app(A-B) The apparent permeability coefficient is measured from the top to the base.

[0310] Recovery rate (Recovery%) is calculated using the following formula:

[0311] in:

[0312] V A The volume of the solution at the receiving end (unit: mL);

[0313] V D The volume of the solution at the administration end (unit: mL);

[0314] [drug] receiver The drug concentration at the receiving end;

[0315] [drug]initial,donor is the initial drug concentration at the dosing end.

[0316] [drug] donor This represents the drug concentration at the dosing end.

[0317] The integrity of the Caco-2 cell monolayer membrane was calculated using the following formula:

[0318] in:

[0319] I receiver This refers to the fluorescence density of the receiving aperture (0.3 mL).

[0320] I donor This refers to the fluorescence density of the drug delivery well (0.1 mL).

[0321] Integrity is expressed as LY Leakage percentage (%LY Leakage).

[0322] %LY Leakage < 1.5% indicates an intact monolayer cell membrane. For individual cases where %LY Leakage > 1.5%, if P... app The values ​​are similar to those of other parallel holes, and based on scientific judgment, the final data can be adopted.

[0323] Table 4. Permeability coefficients of test substances in Caco-2 cell models

[0324] Experimental results showed that the efflux rates of compounds 1, 3, 6, 10 and 26 were significantly lower than those of Dxd, with compound 1 having the lowest efflux rate.

[0325] In addition to those described herein, various modifications of the invention will be apparent to those skilled in the art based on the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. All references cited in this application (including all patents, patent applications, journal articles, books, and any other disclosures) are incorporated herein by reference in their entirety.

Claims

1. A compound having the structure of Formula (I): or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, wherein: R 1 and R 2 are independently selected from the group consisting of hydroxy, halogen, -C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, -C 1-3 alkyl-NR a R b or -NR a R b or, alternatively, R 1 , R 2 and the adjacent C atoms form a 5-6 membered heterocycloalkyl group; R 3 selected from H, C 1-3 alkyl or C 1-3 haloalkyl; R 4 selected from C 1-6 haloalkyl, C 1-6 alkyl or C 3-10 cycloalkyl, and said R 4 is further substituted by R 4A ; R 4A selected from hydroxyl, thiol, or -NR 4A-1 -(CH2) m -R 4A-2 , and said R 4A optionally substituted with halogen at the sites that can be substituted; R 4A-1 selected from H, C 1-3 alkyl, C 1-3 haloalkyl or C 1-3 alkoxy; R 4A-2 H, halogen, =0, CN, -S(=0)2-C 1-3 alkyl, -S(=0)2-C 4-6 alkyl, C 3-6 cycloalkyl, 5-6 membered heterocycloalkyl, C 6-10 aryl or 5-7 membered heteroaryl; or R 3 , R 4 and the adjacent C atom form a 5-10 membered cycloalkyl, and said 5-10 membered cycloalkyl is optionally substituted with one or more R x ; R x independently selected from halogen, NR c R d , hydroxy, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl or C 1-3 alkoxy; R a and R b are independently selected from H, C 1-3 alkyl or C 1-3 haloalkyl; R c and R d are independently selected from H or -COCH3; n is 0 or 1 ; m is 0, 1, 2, 3 or 4; when n is 0, and R 3 , R 4 forms a 5-10 membered cycloalkyl with the adjacent C atom, the 5-10 membered cycloalkyl is cyclohexyl, and the 5-10 membered cycloalkyl is substituted with -NHAc, R 1 and R 2 are independently selected from halogen; or, the 5-10 membered cycloalkyl is cyclohexyl, and the 5-10 membered cycloalkyl is optionally substituted with halogen, R 1 is hydroxyl, halogen, C 1-6 haloalkyl, -C 1-3 alkyl-NR a R b or C 1-6 alkoxy, R 2 is halogen; or, the 5-10 membered cycloalkyl is cyclohexyl, R 1 is hydroxyl, R 2 is halogen; or, the 5-10 membered cycloalkyl is a 5-10 membered bridged cycloalkyl, and the 5-10 membered cycloalkyl is optionally substituted with one or more R x ; or, the 5-10 membered cycloalkyl is a partially unsaturated 5-10 membered cycloalkyl, and the 5-10 membered cycloalkyl is optionally substituted with one or more R x ; when n is 1, and R 3 , R 4 and the adjacent C atom form a 5-10 membered cycloalkyl group, R 1 and R 2 are independently selected from the group consisting of hydroxyl, halogen, -C 1-6 alkyl, C 1-6 haloalkyl or C 1-6 alkoxy, or R 1 , R 2 and the adjacent C atom form a 5-6 membered heterocycloalkyl group; When R 4 C 1-6 When alkyl, n is 1 and R 1 Selected from hydroxyl, halogen, -C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 alkoxy or -C 1-3 Alkyl-NR a R b .

2. The compound of claim 1, wherein R x is independently selected from halogen, NR c R d , C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, or C 1-3 alkoxy.

3. The compound of claim 1 or 2, wherein: R 4 selected from C 1-6 haloalkyl, C 1-6 alkyl or C 3-10 cycloalkyl, and said R 4 is further substituted by R 4A R 4A is selected from hydroxyl or -NR 4A-1 -(CH2) m -R 4A-2 , and said R 4A is optionally substituted with halogen at the sites where substitution is possible; R 4A-2 is selected from H, halogen, =0, CN, -S(=0)2-C 1-3 alkyl, C 3-6 cycloalkyl, 5-6 membered heterocycloalkyl, C 6-10 aryl or 5-7 membered heteroaryl; Preferably, wherein R 4A-1 is H; Preferably, said R 4 is selected from C 1-6 haloalkyl, C 1-6 alkyl, or C 3-10 cycloalkyl, said R 4 is further substituted by hydroxy, and R c and R d are independently selected as H.

4. The compound of any one of claims 1 to 3, wherein the compound of Formula (I) is a compound of Formula (I-l): wherein: R 1 selected from the group consisting of hydroxyl, halogen, C 1-6 haloalkyl, -C 1-3 alkyl-NR a R b or C 1-6 alkoxy; preferably, R 1 is selected from the group consisting of hydroxyl, F, CI, Br, I, -CF3, -CHF2, -CH2F, -CF2CH3, -CF2CH2CH3, -CH2CF3, -CH2CF2CH3, -CH2CH2CF3, -CH2-N(CH3)2, -CH2-NH2, methoxy, ethoxy, or propoxy, further preferred hydroxyl, F, -CH2-N(CH3)2, -CH2-NH2, or methoxy; R 2 selected from halogen; R L optionally selected from halogen, preferably halogen, more preferably F; and m is 2.

5. The compound of claim 1, wherein the compound of Formula (I) is a compound of Formula (I-2): ###00010### (I-2) ​ wherein: R 1 and R 2 are independently selected from the group consisting of hydroxyl, halogen, C 1-6 haloalkyl, -C 1-3 alkyl-NR a R b , -NR a R b , or C 1-6 alkoxy; preferably, R 1 and R 2 are independently selected from the group consisting of hydroxyl, F, Cl, Br, I, -CF3, -CHF2, -CH2F, -CF2CH3, -CF2CH2CH3, -CH2CF3, -CH2CF2CH3, -CH2CH2CF3, -CH2-N(CH3)2, -NH2, methoxy, ethoxy, or propoxy, more preferably hydroxyl, F or NH2, further preferably, R 1 is hydroxyl, F or NH2, and R 2 is F; R L selected from halogen, -OH, C 1-3 alkyl or C 1-3 haloalkyl; and m is 0, 1, 2 or 3.

6. The compound of any one of claims 1-3, wherein the compound of Formula (I) is a compound of Formula (I-3): wherein: R 1 and R 2 are independently selected from halogen; preferably, R 1 and R 2 are independently selected from F, CI, Br or I, more preferably F or CI; further preferably, R 1 is F or CI, and R 2 is F; R L selected from -NHAc; and m is 1.

7. The compound of any one of claims 1- to 3, wherein the compound of Formula (I) is a compound of Formula (I-4): wherein: R 3 selected from H; R 4 selected from C 1-6 haloalkyl or C 3-10 cycloalkyl, and said R 4 is further substituted by R 4A substituted; R 1 selected from C 1-3 alkyl or C 1-3 alkoxy, preferably methyl or methoxy; R 2 selected from F, CI, Br or I, preferably F; Or, the R 1 R 2 It forms a 5-6 membered heterocyclic alkyl group with adjacent C atoms; preferably, the R 1 R 2 Composed of adjacent C atoms 8. The compound of claim 7, wherein: said R 4A selected from hydroxyl or -NH-(CH2) m -R 4A-2 and said R 4A optionally substituted with halogen at the sites where substitution can occur; and / or R 4 selected from C 1-6 haloalkyl, and said R 4 further substituted by hydroxy or -NH-(CH2) m -R 4A-2 substituted; R is selected from H, F, CI, Br, I, CN, -S(=0)2-CH3, or C 4A-2 R is selected from H, F, CI, Br, I, CN, -S(=0)2-CH3, or C 1-3 R is selected from H, F, CI, Br, I, CN, -S(=0)2-CH3, or C 3-6 R is selected from H, F, CI, Br, I, CN, -S(=0)2-CH3, or C 4A-2 R is selected from H, F, CI, Br, I, CN, -S(=0)2-CH3, or C 3-6 R is selected from H, F, CI, Br, I, CN, -S(=0)2-CH3, or C 3-6 R is selected from H, F, CI, Br, I, CN, -S(=0)2-CH3, or C 9. The compound of claim 7, wherein R 4 is selected from C 1-6 haloalkyl or C 3-10 cycloalkyl, and said R 4 is further substituted with hydroxy.

10. The compound of claim 7, wherein R 4 is selected from the group consisting of -CF2OH, -CF2CH2OH, -CF2CH2CH2OH, -CH2CF2OH, -CH2CF2CH2OH, -CH2CH2CF2OH, hydroxy-substituted cyclobutyl, hydroxy-substituted cyclopentyl, hydroxy-substituted cyclohexyl, is preferably -CF2OH, -CF2CH2OH, -CF2CH2CH2OH, -CH2CF2OH, -CH2CF2CH2OH, -CH2CH2CF2OH, hydroxy-substituted cyclobutyl, hydroxy-substituted cyclopentyl, hydroxy-substituted cyclohexyl, Further preferably, R 4 is selected from the group consisting of -CF2OH, -CF2CH2OH, -CF2CH2CH2OH, -CH2CF2OH, -CH2CF2CH2OH, -CH2CH2CF2OH, hydroxy-substituted cyclobutyl, hydroxy-substituted cyclopentyl, or hydroxy-substituted cyclohexyl, more preferably hydroxy-substituted cyclobutyl, hydroxy-substituted cyclopentyl, or hydroxy-substituted cyclohexyl.

11. The compound of any one of claims 1 to 3, wherein the compound of Formula (I) is a compound of Formula (I-5): wherein: R 4 selected from C 1-6 haloalkyl or C 1-6 alkyl, and said R 4 further substituted by hydroxy; preferably, said R 4 selected from -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, -CF2OH, -CF2CH2OH, -CF2CH2CH2OH, -CH2CF2OH, -CH2CF2CH2OH, -CH2CH2CF2OH, Further preferred R 1 and R 2 form, together with the adjacent C atoms, a 5-6 membered heterocycloalkyl group; preferably, R 1 and R 2 form, together with the adjacent C atoms, a 12. The compound of claim 1, wherein the compound of Formula (I) is a compound of Formula (I-6): ###00035### (I-6) ​ wherein: R 1 and R 2 are independently selected from the group consisting of hydroxy, halogen, -C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, -C 1-3 alkyl-NR a R b , or -NR a R b , or, R 1 , R 2 and the adjacent C atom form a 5-6 membered heterocycloalkyl group; R L selected from halogen, NR c R d , -OH, C 1-3 alkyl, C 1-3 alkoxy or C 1-3 haloalkyl, preferably halogen, -OH, C 1-3 alkyl, C 1-3 alkoxy or C 1-3 haloalkyl; and m is 0, 1, 2 or 3, preferably 2 or 3.

13. The compound of claim 1, wherein the compound of Formula (I) is a compound of Formula (I-7): ###00036### (I-7) ​ wherein: R 1 and R 2 are independently selected from the group consisting of hydroxy, halogen, -C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, -C 1-3 alkyl-NR a R b , or -NR a R b , or, R 1 , R 2 and the adjacent C atom form a 5-6 membered heterocycloalkyl group; R L selected from halogen, -OH, C 1-3 alkyl, C 1-3 alkoxy or C 1-3 haloalkyl; and m is 0, 1, 2 or 3.

14. The compound of claim 1, wherein the compound of Formula (I) is the following compound:

15. A pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound of any one of claims 1-14, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, and one or more pharmaceutically acceptable carriers.

16. Use of a compound of any one of claims 1-14, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, or a pharmaceutical composition of claim 15, for the manufacture of a medicament for the prevention or treatment of cancer and / or tumors and related conditions thereof.

17. A compound of any one of claims 1-14, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, or a pharmaceutical composition of claim 15, for use in the prevention or treatment of cancer and / or tumors and related conditions thereof.

18. A method of preventing or treating cancer and / or tumors and related conditions thereof, comprising administering to an individual in need thereof an effective amount of a compound of any one of claims 1-14, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, or a pharmaceutical composition of claim 15.

19. The method of claim 18, wherein the cancer and / or tumors and related conditions thereof are selected from the group consisting of cancers arising in the esophagus, stomach, intestine, rectum, mouth, pharynx, larynx, lung, colon, breast, uterus, endometrium, ovary, prostate, testis, bladder, kidney, liver, pancreas, bone, connective tissue, skin, eye, brain, and central nervous system, as well as thyroid cancer, leukemia, Hodgkin's disease, lymphoma, and myeloma.

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