Tricyclic compounds, methods for preparing same, and uses thereof

Abstract

The present invention belongs to the field of medicine, and particularly relates to a tricyclic compound, a method for preparing the same and its use.In particular, the present invention relates to a compound represented by formula (I) or its pharma-ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug, a method for preparing the same, a pharmaceutical composition containing the same, and their use. TIFF2025501049000118.tif52149

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is based on and claims priority from Chinese Patent Application No. 202111666009.2, filed on December 30, 2021, and Chinese Patent Application No. 202210467340.X, filed on April 29, 2022. The disclosures of these Chinese patent applications are incorporated herein in their entireties. [Technical field]

[0002] The present invention is in the field of medicinal chemistry and in particular relates to tricyclic compounds or their pharma-ceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs, processes for their preparation, pharmaceutical compositions containing them and their uses. [Background technology]

[0003] Toll-like receptors (TLRs) belong to the pattern recognition receptor (PRR) family, which can specifically recognize pathogen-associated molecular patterns (PAMPs). They are an important class of proteins involved in non-specific immunity and are the bridge between non-specific and specific immunity. Traditionally, 13 types of Toll-like receptors have been found in mammals, and 10 types (TLR1-10) have been found in humans, among which TLR1 / 2 / 4 / 5 / 6 / 10 are expressed on the cell surface and can rapidly recognize the products of bacterial metabolism, and TLR3 / 7 / 8 / 9 are expressed in endosomes and mainly recognize and monitor exogenous nucleic acid substances within pathogen cells.

[0004] TLR7 has a limited expression distribution in humans, and is mainly expressed by B cells and plasmacytoid dendritic cells (pDCs). In response to viral infection, TLR7 induces high levels of interferon-α, enhances the activation of pDCs' costimulation and antigen-presenting ability, promotes the proliferation of CD4+ T cells, and further activates CD8+ T cells to generate adaptive T cell responses. The expression of TLR8 is relatively more widespread, and is mainly expressed by monocytes, NK cells, and myeloid dendritic cells (mDCs) in humans. Stimulation of TLR8 can induce the release of various inflammatory cytokines, such as tumor necrosis factor α (TNF-α), IL-6, IL-12, and interferon-γ. At the same time, it has also been proven that the TLR8 signaling pathway is a necessary and sufficient condition for reversing the suppressive function of Tregs cells. Therefore, TLR7 and TLR8 play important roles in the innate and adaptive immunity of the organism, and are common targets for anti-cancer immunotherapy and anti-viral immunotherapy. Although several TLR7 and / or TLR8 agonists have been reported by different companies at present, there is still a great demand for TLR7 and / or TLR8 agonists with better activity, safety and physicochemical properties.

[0005] Currently, a new type of immunostimulatory antibody conjugate (ISAC) has emerged by linking immunomodulators, such as TLR7 and / or TLR8 agonists, to tumor-targeting antibodies via cleavable / non-cleavable linkers. Immunostimulatory antibody conjugates combine the precise tumor localization of antibodies with the sustained antitumor activity and immune memory efficacy of immunomodulators, such as TLR7 and / or TLR8 agonists, to safely induce antitumor immune responses while allowing systemic administration.

[0006] Therefore, in view of the above aspects, the present invention provides a class of tricyclic compounds with TLR7 and / or TLR8 agonist activity. This type of compound has excellent activity, safety and physical and chemical properties, and can be further used in immunostimulatory antibody conjugates, which is of great significance for the development of antitumor and antiviral drugs.

[0007] In one aspect, the present application provides a class of tricyclic compounds that have strong agonistic effects on TLR7 and / or TLR8, and thus are better suited for use in the prevention and / or treatment of diseases associated with the mediation of TLR7 and / or TLR8. The compounds also have various excellent properties, such as good physicochemical properties (e.g., solubility, physical and / or chemical stability) and good safety. The compound has the formula (I):

[0008] [ka] (In the formula, X 1 is selected from the group consisting of N and C; X 2 is selected from the group consisting of N and C; X 1 and X 2 at least one of is selected from N; X 3 is selected from the group consisting of O, S and N; X 4 are O, S, N and CR 4 selected from the group consisting of; R 1 is C 1~6 Alkyl and -C 1~6 Alkylene-OC 1~6 selected from the group consisting of alkyl; R 2 is hydrogen or -L 1 -L 2 -L 3 -L 4 having the formula: L 1 is a covalent bond and C 1~6 alkylene; L 2 is a covalent bond, C 3~10 Cycloalkyl, 3-12 membered heterocyclyl, C 6~10aryl and 5-10 membered heteroaryl, optionally wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C 1~6 Alkyl, C 1~6 Alkoxy and -C 1~6 Alkylene-NH 2 each independently substituted with one or more groups independently selected from the group consisting of: L 3 is a covalent bond, -O-, -S-, -C(O)-, -OC(O)-, -C(O)-O-, -OC(O)-O-, -NR 5 -, -C(O)-NR 5 -, -NR 5 -C(O)-, -OC(O)-NR 5 -, -NR 5 -C(O)-O-, -NR 5 -C(O)-NR 5 -, -S(O) m -NR 5 -, -NR 5 -S(O) m - and -NR 5 -S(O) m -NR 5 - selected from the group consisting of; L 4 is hydrogen, C 1~6 Alkyl, -(O-CH 2 CH 2 -) n O.C. 1~6 Alkyl and -C 1~6 Alkylene-(O-CH 2 CH 2 -) n O.C. 1~6 alkyl, optionally each alkyl group is selected from the group consisting of hydrogen, halogen, hydroxyl, -NH 2 , -NH-C(O)-OC 1~6 Alkyl, C 1~6 Alkoxy, carboxyl and -C(O)-OC 1~6 independently substituted with one or more groups independently selected from the group consisting of alkyl; R 3 is hydrogen or -L 5-L 6 -L 7 -L 8 having the formula: L 5 is a covalent bond and C 1~6 alkylene; L 6 is a covalent bond, C 3~10 Cycloalkyl, 3-12 membered heterocyclyl, C 6~10 aryl and 5-10 membered heteroaryl, optionally wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C 1~6 Alkyl, C 1~6 Alkoxy and -C 1~6 Alkylene-NH 2 each independently substituted with one or more groups independently selected from the group consisting of: L 7 is a covalent bond, -O-, -S-, -C(O)-, -OC(O)-, -C(O)-O-, -OC(O)-O-, -NR 6 -, -C(O)-NR 6 -, -NR 6 -C(O)-, -OC(O)-NR 6 -, -NR 6 -C(O)-O-, -NR 6 -C(O)-NR 6 -, -S(O) p -NR 6 -, -NR 6 -S(O) p - and -NR 6 -S(O) p -NR 6 - selected from the group consisting of; L 8 is hydrogen, C 1~6 Alkyl, -(O-CH 2 CH 2 -) q O.C. 1~6 Alkyl and -C 1~6 Alkylene-(O-CH 2 CH 2 -) q O.C. 1~6alkyl, optionally each alkyl group is selected from the group consisting of hydrogen, halogen, hydroxyl, -NH 2 , -NH-C(O)-OC 1~6 Alkyl, C 1~6 Alkoxy, carboxyl and -C(O)-OC 1~6 substituted with one or more groups independently selected from the group consisting of alkyl; R 4 is hydrogen, halogen, cyano, hydroxyl, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxy and C 3~10 cycloalkyl; Each R 5 is hydrogen and C 1~6 independently selected from the group consisting of alkyl; Each R 6 is hydrogen and C 1~6 independently selected from the group consisting of alkyl; each m is independently 1 or 2; Each n is independently selected from the group consisting of integers 1 to 25; each p is independently 1 or 2; and each q is independently selected from the group consisting of integers 1 to 25), or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

[0009] The present application further provides the following drug-linkers: D-L' wherein D is a fragment of a compound described in any one of the first aspects; and L is a linker), or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

[0010] The present application also provides a compound of formula (ISAC-I): [ka] (In the formula, D is a fragment of a compound described in any one of the first aspects; L is a fragment of the linker L', where L' is as defined in the second aspect; The Ab is an antibody capable of targeting a target antigen, optionally the antibody is modified (e.g., modified with a cross-linking agent); and z is selected from the group consisting of 1 to 10), or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

[0011] In another aspect, the present application provides the use of a compound of Formula (I) in the manufacture of a drug-linker compound.

[0012] In another aspect, the present application provides the use of a compound of formula (I) and / or a drug-linker in the preparation of an immunostimulatory antibody conjugate.

[0013] In another aspect, the present application provides a method for preparing an immunostimulatory antibody conjugate, the method comprising using a compound of formula (I) and / or a drug-linker.

[0014] In another aspect, the application provides pharmaceutical compositions comprising a prophylactically and / or therapeutically effective amount of a compound described herein or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, or a drug-linker or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, or an immunostimulatory antibody conjugate described in formula (ISAC-I), and one or more pharma- ceutically acceptable carriers.

[0015] In another aspect, the application provides a kit comprising a compound of formula (I) or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, or a Drug-Linker or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, or an immunostimulatory antibody conjugate of formula (ISAC-I), or a pharmaceutical composition.

[0016] In another aspect, the present application provides the use of a compound of formula (I) or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, or an immunostimulatory antibody conjugate of formula (ISAC-I), or a drug-linker or a pharma- ceutically acceptable salt, ester, stereoisomer, isomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, or a pharmaceutical composition, in the manufacture of a medicament for the prevention and / or treatment of a disease mediated by TLR7 and / or TLR8 (e.g., a tumor).

[0017] In another aspect, the present application provides a method for preventing and / or treating a disease mediated by TLR7 and / or TLR8 (e.g., tumors), comprising administering to a subject in need thereof an effective amount of a compound of formula (I) or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, or an immunostimulatory antibody conjugate of formula (ISAC-I), or a drug-linker or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, or a pharmaceutical composition.

[0018] In another aspect, the application provides methods of preparing the compounds described herein.

[0019] definition Unless otherwise defined below, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. References to technology used herein are intended to refer to technology commonly understood in the art, including modifications of the technology or equivalent substitutions that are obvious to those skilled in the art. The following terms are believed to be well understood by those skilled in the art, and the following definitions are provided to better explain the present invention.

[0020] As used herein, the terms "comprise," "include," "have," "contain," or "involve," and other variations thereof, are inclusive or open-ended and do not exclude other unrecited elements or method steps.

[0021] As used herein, the term "alkyl" is defined as a straight or branched chain saturated aliphatic hydrocarbon group. For example, as used herein, the term "C 1~6 "Alkyl" refers to a straight or branched chain group having from 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, or n-hexyl) optionally substituted with one or more (e.g., 1 to 3) suitable substituents such as halogen. 1~3 "Alkyl" refers to a straight or branched chain group having 1 to 3 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl) optionally substituted with one or more (e.g., 1 to 3) suitable substituents such as halogen.

[0022] As used herein, the term "alkoxy" refers to "alkyl-O-", where "alkyl" is as defined above. For example, the term "C 1~6 Alkoxy means "C 1~6 "C 1~6 The term "alkyl" is as defined above. 1~3 Alkoxy means "C 1~3"C 1~3 "Alkyl" is as defined above. Exemplary C 1~6 Alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy or n-hexyloxy.

[0023] As used in the present invention, "C 1~6 "Alkylene" refers to a divalent group formed by removing two hydrogens from a straight-chain or branched alkane containing 1 to 6 carbon atoms; 1~5 Alkylene", "C 1~4 Alkylene", "C 1~3 Alkylene", "C 1~2 Specific examples include, but are not limited to, -CH 2-、 -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 --etc.

[0024] As used herein, the term "cycloalkyl" refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic (e.g., bicyclic) hydrocarbon ring (e.g., a monocyclic ring such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or a bicyclic ring containing a spiro, fused, or bridged system such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, or bicyclo[5.2.0]nonyl, decalinyl, etc.), optionally substituted with one or more (e.g., 1-3) suitable substituents. For example, the term "C 3~6 "Cycloalkyl" refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic (e.g., bicyclic) hydrocarbon ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), which is optionally substituted with one or more (e.g., 1 to 3) suitable substituents, such as cyclopropyl substituted with methyl.

[0025] As used herein, the term "halogen" group is defined to include fluorine, chlorine, bromine, or iodine.

[0026] As used herein, the term "halo" refers to substitution with one or more (eg, 1 to 3) of the same or different halogen atoms.

[0027] As used herein, the term "haloalkyl" refers to an alkyl substituted with one or more (e.g., 1 to 3) of the same or different halogen atoms. For example, the term "C 1~6 "Haloalkyl" refers to haloalkyl having 1 to 6 carbon atoms, e.g., -CF 3 , -C 2 F 5 , -CHF 2 , -CH 2 F, -CH 2 CF 3 , -CH 2 Cl or -CH 2 CH 2 CF 3 Etc.

[0028] As used herein, the term "heterocycle" or "heterocyclyl" refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic ring group, e.g., having 2, 3, 4, 5, 6, 7, 8, or 9 carbon atoms and N, O, or S(O) in the ring. t (wherein t is 0, 1 or 2), one or more (e.g., 1, 2, 3 or 4) heteroatoms independently selected from the group consisting of, for example, 3- to 12-membered heterocyclyl, 3- to 7-membered heterocyclyl, 3- to 6-membered heterocyclyl, 5- to 6-membered heterocyclyl, etc., for example, 5- to 10-membered nitrogen-containing heterocyclyl, 6- to 10-membered oxygen-containing heterocyclyl, 6- to 8-membered sulfur-containing heterocyclyl, 5- to 8-membered oxygen-containing heterocyclyl, etc. Representative examples of heterocyclyl include, but are not limited to, oxiranyl, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuryl, tetrahydropyrrolidinyl, hexahydro-1H-pyrrolinyl, pyrrolidonyl, imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, tetrahydropyridyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, and the like.

[0029] As used herein, the term "aryl" or "aromatic ring" refers to an all-carbon monocyclic or fused ring polycyclic aromatic group having a conjugated pi-electron system. 6~10 Aryl" or "C 6~10 "Aromatic ring" refers to an aromatic group containing 6 to 10 carbon atoms, such as a phenyl (ring) or naphthyl (ring). Aryl may have one or more (e.g., 1 to 3) suitable substituents (e.g., halogen, -OH, -CN, -NO 2 , C 1~6 The aryl group is optionally substituted with an alkyl group, such as an alkyl group.

[0030] As used herein, the term "heteroaryl" refers to an aromatic cyclic group in which at least one ring atom is a heteroatom, such as a nitrogen atom, an oxygen atom, or a sulfur atom. Optionally, a ring atom (e.g., a carbon, nitrogen, or sulfur atom) in the ring structure may be replaced with oxygen. Specific examples include, but are not limited to, 5-10 membered heteroaryl, 5-10 membered nitrogen-containing heteroaryl, 6-10 membered oxygen-containing heteroaryl, 6-8 membered nitrogen-containing heteroaryl, 5-8 membered oxygen-containing heteroaryl, and the like, such as furyl, thienyl, pyrrolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5- Examples of such aryl groups include oxadiazolyl, 1,3,4-oxadiazolyl, pyridyl, 2-pyridonyl, 4-pyridonyl, pyrimidinyl, 1,4-dioxinyl, 2H-1,2-oxazinyl, 4H-1,2-oxazinyl, 6H-1,2-oxazinyl, 4H-1,3-oxazinyl, 6H-1,3-oxazinyl, 4H-1,4-oxazinyl, pyridazinyl, pyrazinyl, 1,2,3-triazinyl, 1,3,5-triazinyl, 1,2,4,5-tetrazinyl, azepinyl, 1,3-diazepinyl, azacyclooctatetraenyl, and the like.

[0031] Optionally, hydrogen in groups referred to in the present invention may be replaced with deuterium.

[0032] The term "substituted" means that one or more (e.g., 1, 2, 3, or 4) hydrogens on the designated atom are selectively replaced with the indicated group, provided that the designated atom has a valence that does not exceed its normal valence under the present circumstances, and such substitution results in a stable compound. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0033] When a substituent is described as "optionally substituted with," the substituent may be (1) unsubstituted or (2) substituted. When a carbon of a substituent is described as being optionally substituted with one or more of the enumerated substituents, the hydrogen or hydrogens bearing that carbon (to the extent of any hydrogens present) may or may not be substituted, independently and / or together, with the selected substituent. When a nitrogen of a substituent is described as being optionally substituted with one or more of the enumerated substituents, the hydrogen or hydrogens bearing the nitrogen (to the extent of any hydrogens present) may or may not be substituted, independently, with the selected substituent.

[0034] When substituents are described as "independently selected from" a set of groups, each substituent is selected independently from the other groups. Thus, each substituent can be the same or different from another (other) substituent.

[0035] As used herein, the term "one or more" means, for example, one or more than one under reasonable conditions, such as 2, 3, 4, 5, 6, 7, 8, 9 or 10. As used herein, unless otherwise indicated, the site of attachment of a substituent may be at any suitable position of the substituent. When a bond to a substituent is shown to be via a bond connecting two atoms in a ring, then such substituent may be bonded to any ring-forming atom in such substitutable ring.

[0036] The present invention also includes all pharma- ceutically acceptable isotopically labeled compounds that are identical to the compounds described herein except that one or more atoms are replaced with an atom having the same atomic number but a different atomic mass or mass number than the atom predominant in nature. Examples of isotopes suitable for inclusion in the compounds include, but are not limited to, isotopes of hydrogen (e.g., 2 H, 3 H, deuterium D, tritium T); isotopes of carbon (e.g., 11 C. 13 C, and 14C); isotopes of chlorine (e.g., 37 Cl); isotopes of fluorine (e.g., 18 F), isotopes of iodine (e.g. 123 I and 125 I), nitrogen isotopes (e.g., 13 N and 15 N), isotopes of oxygen (e.g. 15 O. 17 O and 18 O), phosphorus isotopes (e.g. 32 P), sulfur isotopes (e.g. 35 Certain isotopically labeled compounds of the present invention (e.g., those incorporating a radioactive isotope) are useful in drug and / or substrate tissue distribution studies (e.g., assays). The radioactive isotope tritium (i.e., 3 H) and carbon-14 (i.e. 14 C) are particularly useful for this purpose due to their ease of incorporation and detection. Positron emitting isotopes, e.g. 11 C. 18 F, 15 O, and 13 Substitution with N, etc., 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 schemes and / or examples, and by substituting the appropriate isotopically labeled reagent for the previously used non-labeled reagent. Pharmaceutically acceptable solvates of the present invention include those that are compatible with solvents of crystallization, e.g., D 2 O, acetone-d6 or DMSO-d 6 etc., which can be isotopically substituted.

[0037] The term "stereoisomer" refers to an isomer formed as a result of at least one asymmetric center. In the case of compounds having one or more (e.g., 1, 2, 3, or 4) asymmetric centers, racemic mixtures, single enantiomers, diastereomeric mixtures, and single diastereoisomers may occur. Some individual molecules may also exist as geometric isomers (cis / trans). Similarly, the compounds of the present invention may exist as mixtures of two or more structurally distinct forms (commonly referred to as tautomers) that are in rapid equilibrium. Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, and the like. It is to be understood that the scope of the present application encompasses all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

[0038] The present invention includes all possible crystalline forms, or polymorphs, of the compounds of the present invention, which may be a single polymorph or a mixture of two or more polymorphs in any ratio.

[0039] It should also be understood that some compounds of the present invention may exist in free form for use in therapy, or, where appropriate, as their pharmaceutically acceptable derivatives. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, solvates, metabolites or prodrugs, which can directly or indirectly provide the compound of the present invention or its metabolites or residues. Thus, when "compounds of the present invention" are referred to herein, it is also intended to include the aforementioned various derivative forms of the compound.

[0040] Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts. Suitable acid addition salts are formed from acids capable of forming pharma-ceutically acceptable salts. Suitable base addition salts are formed from bases capable of forming pharma-ceutically acceptable salts. For a review of suitable salts, see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" (Wiley-VCH, 2002). Methods for preparing pharma-ceutically acceptable salts of the compounds of the present invention are known to those skilled in the art.

[0041] The compounds of the present invention can exist in the form of solvates, preferably hydrates, which contain polar solvents as structural elements of the crystal lattice of the compounds. The amount of polar solvent, in particular water, can be present in a stoichiometric or non-stoichiometric ratio.

[0042] Those skilled in the art understand that not all nitrogen-containing heterocycles are capable of forming N-oxides, since nitrogen requires an available lone pair to oxidize and form an oxide.Those skilled in the art recognize that tertiary amines are capable of forming N-oxides.Synthetic methods for the preparation of N-oxides of heterocycles and tertiary amines are well known to those skilled in the art, and include the use of peroxyacids, such as peracetic acid and m-chloroperbenzoic acid (MCPBA), hydrogen peroxide, alkyl hydroperoxides, such as t-butyl hydroperoxide, sodium perborate, and dioxiranes, such as dimethyldioxirane, to oxidize heterocycles and tertiary amines. These methods for preparing N-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. Cheeseman and E. S. G. Werstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp 390-392, A. R. Katritzky and A. J. Boulton, eds., Academic Press.

[0043] Also included within the scope of the invention are metabolic products of the compounds of the invention, i.e., substances formed in vivo following administration of a compound of the invention. Such products may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, and the like, of the administered compound. Thus, the invention includes metabolic products of the compounds of the invention, including compounds produced by contacting a compound of the invention with a mammal for a period of time sufficient to produce those metabolic products.

[0044] The present invention further includes within its scope prodrugs of the compounds of the present invention, which are some derivatives of the compounds of the present invention, which may have little or no pharmacological activity themselves, and which, when administered to or on the body, can be converted, for example, by hydrolytic cleavage, to the compounds of the present invention having the desired activity. Typically, such prodrugs are functional derivatives of the compounds, which are easily converted in vivo to compounds having the desired therapeutic activity. Additional 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), and "Bioreversible Carriers in Drug Design", Pergamon Press, 1987 (ed. EB Roche, American Pharmaceutical Association). Prodrugs of the present invention can be obtained, for example, by replacing appropriate functional groups present in the compounds of the present invention with some moieties known to those skilled in the art as "pro-moieties" (e.g., as described in "Design of Prodrugs", H. Bundgaard (Elsevier, 1985)).

[0045] The present invention also encompasses the compounds of the present invention that contain protective groups.During the process for preparing the compounds of the present invention, it may be necessary and / or desirable to protect any sensitive or reactive groups of the molecules involved, thereby forming chemically protected forms of the compounds of the present invention.This can be achieved by conventional protective groups, such as those described in "Protective Groups in Organic Chemistry", edited by JFW MacOmie, Plenum Press, 1973; and TW Greene & PG M Huts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, all of which are incorporated herein by reference.Protective groups can be removed at a later appropriate stage using methods known in the art.

[0046] The term "about" means within ±10%, preferably within ±5%, and more preferably within ±2% of the value shown in the table.

[0047] The term "conjugate" refers to a substance obtained by linking a small molecule drug with a targeting moiety. In some embodiments of the present invention, the small molecule drug is linked to the targeting moiety via a linker. The linker can be cleaved under a specific environment (e.g., low intracellular pH environment, acidic tumor microenvironment) or specific action (e.g., action of lysosomal proteases), thereby separating the small molecule drug from the targeting moiety.

[0048] The term "linker" refers to a fragment that connects a small molecule drug to a targeting moiety.

[0049] The term "targeting moiety" refers to a portion of a conjugate that is capable of specifically binding to a target (or a portion of a target) on the surface of a cell. Through interaction of the targeting moiety with the target, the conjugate can be delivered to a certain cell population.

[0050] The term "fragment" refers to the remaining portion of a compound molecule after the loss of one or more atoms or groups of atoms (e.g., a hydrogen atom or a hydroxyl group). For example, a "fragment of a compound" refers to the remaining portion of a compound obtained by the loss of a hydrogen atom or a hydroxyl group after a compound described herein has been linked to a linker in an immunostimulatory antibody conjugate.

[0051] The term "crosslinker" refers to a reagent that has the same or different reactive groups at both ends of a molecule that can covalently interact with amino, sulfhydryl, hydroxyl, or maleimide groups on other molecules to produce a crosslinking effect, such as the crosslinker SATA.

[0052] compound The object of the present application is to provide a compound of formula (I):

[0053] [ka] (In the formula, X 1 is selected from the group consisting of N and C; X 2 is selected from the group consisting of N and C; X 1 and X 2 at least one of is selected from N; X 3 is selected from the group consisting of O, S and N; X 4 are O, S, N and CR 4 selected from the group consisting of; R 1 is C 1~6 Alkyl and -C 1~6 Alkylene-OC 1~6 selected from the group consisting of alkyl; R 2 is hydrogen or -L 1 -L 2 -L 3 -L 4 having the formula: L1 is a covalent bond and C 1~6 alkylene; L 2 is a covalent bond, C 3~10 Cycloalkyl, 3-12 membered heterocyclyl, C 6~10 aryl and 5-10 membered heteroaryl, optionally wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C 1~6 Alkyl, C 1~6 Alkoxy and -C 1~6 Alkylene-NH 2 each independently substituted with one or more groups independently selected from the group consisting of: L 3 is a covalent bond, -O-, -S-, -C(O)-, -OC(O)-, -C(O)-O-, -OC(O)-O-, -NR 5 -, -C(O)-NR 5 -, -NR 5 -C(O)-, -OC(O)-NR 5 -, -NR 5 -C(O)-O-, -NR 5 -C(O)-NR 5 -, -S(O) m -NR 5 -, -NR 5 -S(O) m - and -NR 5 -S(O) m -NR 5 - selected from the group consisting of; L 4 is hydrogen, C 1~6 Alkyl, -(O-CH 2 CH 2 ) n -OC 1~6 Alkyl and -C 1~6 Alkylene-(O-CH 2 CH 2 ) n -OC 1~6 alkyl, optionally each alkyl being selected from the group consisting of hydrogen, halogen, hydroxyl, -NH 2 , -NH-C(O)-OC 1~6 Alkyl, C1~6 Alkoxy, carboxyl and -C(O)-OC 1~6 independently substituted with one or more groups independently selected from the group consisting of alkyl; R 3 is hydrogen or -L 5 -L 6 -L 7 -L 8 having the formula: L 5 is a covalent bond and C 1~6 alkylene; L 6 is a covalent bond, C 3~10 Cycloalkyl, 3-12 membered heterocyclyl, C 6~10 aryl and 5-10 membered heteroaryl, optionally wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C 1~6 Alkyl, C 1~6 Alkoxy and -C 1~6 Alkylene-NH 2 each independently substituted with one or more groups independently selected from the group consisting of: L 7 is a covalent bond, -O-, -S-, -C(O)-, -OC(O)-, -C(O)-O-, -OC(O)-O-, -NR 6 -, -C(O)-NR 6 -, -NR 6 -C(O)-, -OC(O)-NR 6 -, -NR 6 -C(O)-O-, -NR 6 -C(O)-NR 6 -, -S(O) p -NR 6 -, -NR 6 -S(O) p - and -NR 6 -S(O) p -NR 6 - selected from the group consisting of; L 8 is hydrogen, C 1~6 Alkyl, -(O-CH 2 CH 2 ) q -OC1~6 Alkyl and -C 1~6 Alkylene-(O-CH 2 CH 2 ) q -OC 1~6 alkyl, optionally each alkyl being selected from the group consisting of hydrogen, halogen, hydroxyl, -NH 2 , -NH-C(O)-OC 1~6 Alkyl, C 1~6 Alkoxy, carboxyl and -C(O)-OC 1~6 independently substituted with one or more groups independently selected from the group consisting of alkyl; R 4 is hydrogen, halogen, cyano, hydroxyl, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxy and C 3~10 cycloalkyl; Each R 5 is hydrogen and C 1~6 independently selected from the group consisting of alkyl; Each R 6 is hydrogen and C 1~6 independently selected from the group consisting of alkyl; each m is independently 1 or 2; Each n is independently selected from the group consisting of integers 1 to 25; each p is independently 1 or 2; and each q is independently selected from the group consisting of integers 1 to 25), or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

[0054] In some embodiments, X 1 is N. In some embodiments, X 1 is C.

[0055] In some embodiments, X 2 is N. In some embodiments, X 2 is C.

[0056] In some embodiments, X 1 and X 2 Exactly one of is N.

[0057] In some embodiments, X 3 is O. In some embodiments, X 3 is S. In some embodiments, X 3 is N.

[0058] In some embodiments, X 4 is O. In some embodiments, X 4 is S. In some embodiments, X 4 is N. In some embodiments, X 4 CR 4 and preferably, X 4 is CH. In some embodiments, R 1 is C 1~4 Alkyl and -C 1~4 Alkylene-OC 1~4 selected from the group consisting of alkyl; Preferably, R 1 is C 1~4 Alkyl and -C 1~2 Alkylene-OC 1~2 In some embodiments, R is selected from the group consisting of alkyl, e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, 2-methoxyethyl, or ethoxymethyl. 1 is n-butyl, 2-methoxyethyl, or ethoxymethyl; preferably, R 1 is n-butyl.

[0059] In some embodiments, R 2 is hydrogen or -L 1 -L 2 -L 3 -L 4 Formula (In the formula, L1 is a covalent bond and C 1~6 alkylene; L 2 is a covalent bond, C 3~10 Cycloalkyl (e.g., C 3~6 Cycloalkyl or C 5~6 cycloalkyl), 3-12 membered heterocyclyl (e.g., 3-6 membered saturated or unsaturated nitrogen-containing monoheterocyclyl such as piperidinyl; 3-6 membered saturated or unsaturated oxygen-containing monoheterocyclyl such as tetrahydropyranyl), C 6~10 aryl (e.g., phenyl) and 5-10 membered heteroaryl (e.g., 5-6 membered nitrogen-containing heteroaryl, 5-6 membered oxygen-containing heteroaryl, 5-6 membered sulfur-containing heteroaryl), optionally wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C 1~6 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), C 1~6 Alkoxy (e.g., methoxy, ethoxy) and -C 1~6 Alkylene-NH 2 (For example, -CH 2 -NH 2 ) independently substituted with one or more groups selected from the group consisting of: 2 is a covalent bond, a 3- to 12-membered heterocyclyl (e.g., a 3- to 6-membered saturated or unsaturated nitrogen-containing monoheterocyclyl, such as piperidinyl; a 3- to 6-membered saturated or unsaturated oxygen-containing monoheterocyclyl, such as tetrahydropyranyl) and C 6~10 aryl (e.g., phenyl), and optionally the heterocyclyl and aryl are selected from the group consisting of hydrogen, halogen, cyano, C 1~6 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), C 1~6 Alkoxy (e.g., methoxy, ethoxy) and -C 1~6 Alkylene-NH 2(For example, -CH 2 -NH 2 each independently substituted with one or more groups independently selected from the group consisting of: 2 is selected from the group consisting of a covalent bond, piperidinyl, and tetrahydropyranyl; L 3 is a covalent bond or -C(O)-; L 4 is hydrogen, C 1~6 Alkyl, -(O-CH 2 CH 2 ) n -OC 1~6 Alkyl (e.g., -(O-CH 2 CH 2 ) n -O-methyl, -((O-CH 2 CH 2 ) n -O-ethyl) and -C 1~6 Alkylene-(O-CH 2 CH 2 ) n -OC 1~6 Alkyl (e.g., -CH 2 -(O-CH 2 CH 2 ) n -O-methyl, -CH 2 -CH 2 -(O-CH 2 CH 2 ) n -O-methyl, -CH 2 -(O-CH 2 CH 2 ) n -O-ethyl, -CH 2 -CH 2 -(O-CH 2 CH 2 ) n Optionally, each alkyl (e.g., methyl or ethyl) is selected from the group consisting of hydrogen, halogen, hydroxyl, -NH 2 , -NH-C(O)-OC 1~6 Alkyl (e.g., -NH-C(O)-O-methyl), C 1~6 Alkoxy (e.g., methoxy, ethoxy), carboxyl and -C(O)-OC1~6 alkyl (e.g., —C(O)—O-methyl), and each n is independently selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7; preferably, L 4 is hydrogen, C 1~6 Alkyl and -C 1~6 Alkylene-(O-CH 2 CH 2 ) n -OC 1~6 alkyl, optionally each alkyl being selected from the group consisting of hydroxyl, -NH 2 , C 1~6 Alkoxy, carboxyl and -C(O)-OC 1~6 alkyl, each n is selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7; preferably, L 4 is hydrogen, C 1~4 Alkyl and -C 1~4 Alkylene-(O-CH 2 CH 2 ) n -OC 1~4 alkyl, optionally each alkyl being selected from the group consisting of hydroxyl, -NH 2 , C 1~4 Alkoxy, carboxyl and -C(O)-OC 1~4 and n is selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7.

[0060] In some embodiments, R 2 is hydrogen.

[0061] In some embodiments, R 2 -L 1 -L 2 -L 3 -L 4 It has the formula:

[0062] In some embodiments, L 1 is a covalent bond. In some embodiments, L1 is C 1~6 Alkylene, for example, -CH 2 -, -CH 2 -CH 2 -, -CH 2 -CH 2 -CH 2 -, -CH 2 -CH 2 -CH 2 -CH 2 In some embodiments, L 1 is a covalent bond or -CH 2 -It is.

[0063] In some embodiments, L 2 is a covalent bond.

[0064] In some embodiments, L 2 is C 3~10 Cycloalkyl (e.g., C 3~6 Cycloalkyl or C 5~6 cycloalkyl), 3-12 membered heterocyclyl (e.g., 3-6 membered saturated or unsaturated nitrogen-containing monoheterocyclyl such as piperidinyl; 3-6 membered saturated or unsaturated oxygen-containing monoheterocyclyl such as tetrahydropyranyl), C 6~10 aryl (e.g., phenyl) and 5-10 membered heteroaryl (e.g., 5-6 membered nitrogen-containing heteroaryl, 5-6 membered oxygen-containing heteroaryl, 5-6 membered sulfur-containing heteroaryl), optionally wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C 1~6 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), C 1~6 Alkoxy (e.g., methoxy, ethoxy) and -C 1~6 Alkylene-NH 2 (For example, -CH 2 -NH 2 ) is substituted with one or more groups selected from the group consisting of:

[0065] In some embodiments, L 2 is 3- to 12-membered heterocyclyl (e.g., 3- to 6-membered saturated or unsaturated nitrogen-containing monoheterocyclyl, such as piperidinyl; 3- to 6-membered saturated or unsaturated oxygen-containing monoheterocyclyl, such as tetrahydropyranyl) and C 6~10 aryl (e.g., phenyl), and heterocyclyl and aryl are selected from the group consisting of -C 1~6 Alkylene-NH 2 (For example, -CH 2 -NH 2 ) is optionally replaced.

[0066] In some embodiments, L 3 is a covalent bond. In some embodiments, L 3 is -C(O)-.

[0067] In some embodiments, L 4 is hydrogen, C 1~6 Alkyl, -(O-CH 2 CH 2 ) n -OC 1~6 Alkyl (e.g., -(O-CH 2 CH 2 ) n -O-methyl, -((O-CH 2 CH 2 ) n -O-ethyl) and -C 1~6 Alkylene-(O-CH 2 CH 2 ) n -OC 1~6 Alkyl (e.g., -CH 2 -(O-CH 2 CH 2 ) n -O-methyl, -CH 2 -CH 2 -(O-CH 2 CH 2 ) n -O-methyl, -CH 2 -(O-CH 2 CH 2 ) n -O-ethyl, -CH 2 -CH2 -(O-CH 2 CH 2 ) n n is independently selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7; alkyl (e.g., methyl or ethyl) is selected from the group consisting of hydrogen, halogen, hydroxyl, -NH 2 , -NH-C(O)-OC 1~6 Alkyl (e.g., -NH-C(O)-O-methyl), C 1~6 Alkoxy (e.g., methoxy, ethoxy), carboxyl, and -C(O)-OC 1~6 and optionally substituted with one or more groups selected from the group consisting of alkyl (e.g., -C(O)-O-methyl).

[0068] In some embodiments, L 4 is hydrogen, C 1~6 Alkyl (e.g., methyl, ethyl), -(O-CH 2 CH 2 ) n -OC 1~6 Alkyl (e.g., -(O-CH 2 CH 2 ) n -O-methyl, -((O-CH 2 CH 2 ) n -O-ethyl) and -C 1~6 Alkylene-(O-CH 2 CH 2 ) n -OC 1~6 Alkyl (e.g., -CH 2 -(O-CH 2 CH 2 ) n -O-methyl, -CH 2 -CH 2 -(O-CH 2 CH 2 ) n -O-methyl, -CH 2 -(O-CH 2 CH 2 ) n -O-ethyl, -CH 2 -CH 2 -(O-CH 2 CH2 ) n -O-ethyl), where n is each independently selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7; alkyl (e.g., methyl or ethyl) is selected from the group consisting of hydrogen, hydroxyl, -NH 2 , C 1~6 Alkoxy (e.g., methoxy), carboxyl and -C(O)-OC 1~6 and optionally substituted with one or more groups selected from the group consisting of alkyl (e.g., -C(O)-O-methyl).

[0069] In some embodiments, L 4 is -(O-CH 2 CH 2 ) n -OC 1~6 Alkyl (e.g., -(O-CH 2 CH 2 ) n -O-methyl, -(O-CH 2 CH 2 ) n -O-ethyl) and -C 1~6 Alkylene-(O-CH 2 CH 2 ) n -OC 1~6 Alkyl (e.g., -CH 2 -(O-CH 2 CH 2 ) n -O-methyl, -CH 2 -CH 2 -(O-CH 2 CH 2 ) n -O-methyl, CH 2 -(O-CH 2 CH 2 ) n -O-ethyl, -CH 2 -CH 2 -(O-CH 2 CH 2 ) n n is independently selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7; alkyl (e.g., methyl or ethyl) is selected from the group consisting of hydroxyl, -NH 2, carboxyl and -C(O)-OC 1~6 and optionally substituted with one or more groups selected from the group consisting of alkyl (e.g., -C(O)-O-methyl).

[0070] In some embodiments, R 2 is hydrogen, C 1~6 Alkyl, C 1~6 Alkylene-NH 2 , C 1~6 Alkylene-3 to 12-membered heterocyclyl, C 1~6 Alkylene-C 6~10 Aryl, C 1~6 Alkylene-3 to 12-membered heterocyclyl-C 1~6 Alkylene-(O-CH 2 CH 2 ) n -OC 1~6 Alkyl, C 1~6 Alkylene-3 to 12-membered heterocyclyl-C(O)-C 1~6 Alkylene-(O-CH 2 CH 2 ) n -OC 1~6 alkyl; n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7; 3- to 12-membered heterocyclyl or C 6~10 Aryl is C 1~6 Alkyl, -C 1~6 Alkylene-NH 2 (For example, -CH 2 -NH 2 ) and -C 1~6 Alkylene-methoxy (e.g., -CH 2 CH 2 -OCH 3 ) are optionally substituted; alkyl and heterocyclyl are optionally substituted with hydrogen, hydroxyl, -NH 2 , carboxyl or -C(O)-OC 1~6 In some embodiments, each C is optionally substituted with one or more groups selected from the group consisting of alkyl (e.g., —C(O)—O-methyl). 1~6 Alkylene is independently methylene, ethylene, or butylene. In some embodiments, each C 1~6Alkyl is, independently, methyl or ethyl. In some embodiments, the 3-12 membered heterocyclyl is, for example, a 3-6 membered saturated or unsaturated nitrogen-containing monoheterocyclyl, such as piperidinyl. In some embodiments, the 3-12 membered heterocyclyl is, for example, a 3-6 membered saturated or unsaturated oxygen-containing monoheterocyclyl, such as tetrahydropyranyl. In some embodiments, C 6~10 Aryl is phenyl.

[0071] In some embodiments, R 2 is hydrogen, C 1~6 Alkylene-NH 2 , C 1~6 Alkylene-3 to 12-membered heterocyclyl, C 1~6 Alkylene-C 6~10 Aryl, C 1~6 Alkylene-3 to 12-membered heterocyclyl-C 1~6 Alkylene-(O-CH 2 CH 2 ) n -OC 1~6 Alkyl, C 1~6 Alkylene-3 to 12-membered heterocyclyl-C(O)-C 1~6 Alkylene-(O-CH 2 CH 2 ) n -OC 1~6 alkyl; n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7; 3- to 12-membered heterocyclyl or C 6~10 Aryl is -C 1~6 Alkylene-NH 2 (For example, -CH 2 -NH 2 ) are optionally substituted; alkyl and heterocyclyl are optionally substituted with hydrogen, hydroxyl, -NH 2 , carboxyl or -C(O)-OC 1~6 In some embodiments, each C is optionally substituted with one or more groups selected from the group consisting of alkyl (e.g., —C(O)—O-methyl). 1~6 Alkylene is independently methylene, ethylene, or butylene. In some embodiments, each C 1~6Alkyl is, independently, methyl or ethyl. In some embodiments, the 3-12 membered heterocyclyl is, for example, a 3-6 membered saturated or unsaturated nitrogen-containing monoheterocyclyl, such as piperidinyl. In some embodiments, the 3-12 membered heterocyclyl is, for example, a 3-6 membered saturated or unsaturated oxygen-containing monoheterocyclyl, such as tetrahydropyranyl. In some embodiments, C 6~10 Aryl is phenyl.

[0072] In some embodiments, R 2 is hydrogen, methyl, -CH 2 -CH 2 -CH 2 -CH 2 -NH 2 , -CH 2 -Piperidine, -CH 2 -piperidinyl-methyl, -CH 2 -Tetrahydropyran, -CH 2 -Phenyl-CH 2 -NH 2 , -CH 2 -Piperidinyl-CH 2 -CH 2 -(O-CH 2 CH 2 ) 2 -NH 2 , -CH 2 -Piperidinyl-CH 2 -CH 2 -(O-CH 2 CH 2 ) 6 -O-methyl, -CH 2 -Piperidinyl-C(O)-CH 2 -CH 2 -(O-CH 2 CH 2 ) 2 -NH 2 , -CH 2 -Piperidinyl-C(O)-CH 2 -CH 2 -(O-CH 2 CH 2 ) 7 -NH 2 , -CH 2 -Piperidinyl-C(O)-CH2 -(O-CH 2 CH 2 ) 2 -OH, -CH 2 -Piperidinyl-C(O)-CH 2 -(O-CH 2 CH 2 ) 4 -OH, -CH 2 -Piperidinyl-C(O)-CH 2 -O-CH 2 CH 2 -O-CH 2 -COOH, -CH 2 -Piperidinyl-C(O)-CH 2 -(O-CH 2 CH 2 ) 3 -O-CH 2 -COOH, -CH 2 -Piperidinyl-C(O)-CH 2 -O-CH 2 CH 2 -O-CH 2 -COOCH 3 , -CH 2 -Piperidinyl-C(O)-CH 2 -(O-CH 2 CH 2 ) 3 -O-CH 2 -COOCH 3 , -CH 2 -Piperidinyl-CH 2 -CH 2 -O-CH 3 , -CH 2 -Piperidinyl-CH 2 -CH 2 -O-CH 2 CH 2 -O-CH 3 , -CH 2 -Piperidinyl-CH 2 -CH 2 -(O-CH 2 CH 2 ) 2 -O-CH 3 is selected from the group consisting of:

[0073] In some embodiments, R 2is hydrogen, -CH 2 -CH 2 -CH 2 -CH 2 -NH 2 , -CH 2 -Piperidine, -CH 2 -Tetrahydropyran, -CH 2 -Phenyl-CH 2 -NH 2 , -CH 2 -Piperidyl-CH 2 -CH 2 -(O-CH 2 CH 2 ) 2 -NH 2 , -CH 2 -Piperidinyl-CH 2 -CH 2 -(O-CH 2 CH 2 ) 6 -O-methyl, -CH 2 -Piperidinyl-C(O)-CH 2 -CH 2 -(O-CH 2 CH 2 ) 2 -NH 2 , -CH 2 -Piperidinyl-C(O)-CH 2 -CH 2 -(O-CH 2 CH 2 ) 7 -NH 2 , -CH 2 -Piperidinyl-C(O)-CH 2 -(O-CH 2 CH 2 ) 2 -OH, -CH 2 -Piperidinyl-C(O)-CH 2 -(O-CH 2 CH 2 ) 4 -OH, -CH 2 -Piperidinyl-C(O)-CH 2 -O-CH 2 CH 2 -O-CH 2 -COOH, -CH 2 -Piperidinyl-C(O)-CH2 -(O-CH 2 CH 2 ) 3 -O-CH 2 -COOH, -CH 2 -Piperidinyl-C(O)-CH 2 -O-CH 2 CH 2 -O-CH 2 -COOCH 3 , -CH 2 -Piperidinyl-C(O)-CH 2 -(O-CH 2 CH 2 ) 3 -O-CH 2 -COOCH 3 is selected from the group consisting of: In some embodiments, R 3 is hydrogen or -L 5 -L 6 -L 7 -L 8 Formula (In the formula, L 5 represents a covalent bond and, for example, -CH 2 -, -CH 2 -CH 2 -, -CH 2 -CH 2 -CH 2 -or-CH 2 -CH 2 -CH 2 -CH 2 - C etc. 1~6 alkylene; preferably, L 5 is a covalent bond; L 6 is a covalent bond, C 3~10 Cycloalkyl (e.g., C 3~6 Cycloalkyl or C 5~6 cycloalkyl), 3- to 12-membered heterocyclyl (e.g., 3- to 6-membered saturated or unsaturated nitrogen-containing monoheterocyclyl (e.g., piperidinyl, tetrahydropyridyl (e.g., 1,2,3,6-tetrahydropyridyl), piperazinyl) or 3- to 6-membered saturated or unsaturated oxygen-containing monoheterocyclyl (e.g., tetrahydropyranyl)), C 6~10aryl (e.g., phenyl) and 5-10 membered heteroaryl (e.g., 5-6 membered nitrogen-containing heteroaryl, 5-6 membered oxygen-containing heteroaryl, 5-6 membered sulfur-containing heteroaryl), optionally wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C 1~6 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), C 1~6 Alkoxy (e.g., methoxy, ethoxy) and -C 1~6 Alkylene-NH 2 (For example, -CH 2 -NH 2 ) are each independently substituted with one or more groups selected from the group consisting of: 6 is selected from the group consisting of a covalent bond and a 3- to 12-membered heterocyclyl (e.g., a 3- to 6-membered saturated or unsaturated nitrogen-containing monoheterocyclyl (e.g., piperidinyl, tetrahydropyridyl (e.g., 1,2,3,6-tetrahydropyridyl), piperazinyl) or a 3- to 6-membered saturated or unsaturated oxygen-containing monoheterocyclyl (e.g., tetrahydropyranyl)), where heterocyclyl is selected from the group consisting of hydrogen, halogen, cyano, C 1~6 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), C 1~6 Alkoxy (e.g., methoxy, ethoxy) and -C 1~6 Alkylene-NH 2 (For example, -CH 2 -NH 2 Optionally substituted with one or more groups independently selected from the group consisting of: 6 is selected from the group consisting of a covalent bond, 1,2,3,6-tetrahydropyridyl, piperazinyl, and tetrahydropyranyl; L 7 is a covalent bond or -C(O)-, preferably L 7 is a covalent bond; L 8 is hydrogen, C1~6 Alkyl, -(O-CH 2 CH 2 ) q -OC 1~6 Alkyl (e.g., -(O-CH 2 CH 2 ) q -O-methyl, -(O-CH 2 CH 2 ) q -O-ethyl) and -C 1~6 Alkylene-(O-CH 2 CH 2 ) q -OC 1~6 Alkyl (e.g., -CH 2 -(O-CH 2 CH 2 ) q -O-methyl, -CH 2 -CH 2 -(O-CH 2 CH 2 ) q -O-methyl, -CH 2 -(O-CH 2 CH 2 ) q -O-ethyl, -CH 2 -CH 2 -(O-CH 2 CH 2 ) q Optionally, each alkyl (e.g., methyl or ethyl) is selected from the group consisting of hydrogen, halogen, hydroxyl, -NH 2 , -NH-C(O)-OC 1~6 Alkyl (e.g., -NH-C(O)-O-methyl), C 1~6 Alkoxy (e.g., methoxy, ethoxy), carboxyl and -C(O)-OC 1~6 alkyl (e.g., -C(O)-O-methyl), and each q is independently selected from the group consisting of 1, 2, 3, 4, 5, 6, 7; preferably, L 8 is hydrogen, C 1~6 Alkyl and -C 1~6 Alkylene-(O-CH 2 CH 2 ) q-OC 1~6 alkyl, optionally each alkyl being selected from the group consisting of hydroxyl, -NH 2 , C 1~6 Alkoxy, carboxyl and -C(O)-OC 1~6 alkyl, and q is selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7; preferably, L 8 is hydrogen, C 1~4 Alkyl and -C 1~4 Alkylene-(O-CH 2 CH 2 ) q -OC 1~4 alkyl, optionally each alkyl being selected from the group consisting of one or more C 1~4 alkoxy groups, and q is selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7.

[0074] In some embodiments, R 3 is hydrogen.

[0075] In some embodiments, R 3 is the formula -L 5 -L 6 -L 7 -L 8 It is.

[0076] In some embodiments, L 5 is a covalent bond. In some embodiments, L 5 is C 1~6 Alkylene, for example, -CH 2 -, -CH 2 -CH 2 -, -CH 2 -CH 2 -CH 2 -, -CH 2 -CH 2 -CH 2 -CH 2 -And more.

[0077] In some embodiments, L 6 is a covalent bond.

[0078] In some embodiments, L 6 is C 3~10 Cycloalkyl (e.g., C 3~6 Cycloalkyl or C 5~6 cycloalkyl), 3- to 12-membered heterocyclyl (e.g., 3- to 6-membered saturated or unsaturated nitrogen-containing monoheterocyclyl (e.g., piperidinyl, tetrahydropyridyl (e.g., 1,2,3,6-tetrahydropyridyl), piperazinyl) or 3- to 6-membered saturated or unsaturated oxygen-containing monoheterocyclyl (e.g., tetrahydropyranyl)), C 6~10 aryl (e.g., phenyl) and 5-10 membered heteroaryl (e.g., 5-6 membered nitrogen-containing heteroaryl, 5-6 membered oxygen-containing heteroaryl, 5-6 membered sulfur-containing heteroaryl), wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C 1~6 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), C 1~6 Alkoxy (e.g., methoxy, ethoxy) and -C 1~6 Alkylene-NH 2 (For example, -CH 2 -NH 2 ) is optionally substituted with one or more groups selected from the group consisting of:

[0079] In some embodiments, L 6 is C 3~10 Cycloalkyl (e.g., C 3~6 Cycloalkyl or C 5~6 cycloalkyl), 3- to 12-membered heterocyclyl (e.g., 3- to 6-membered saturated or unsaturated nitrogen-containing monoheterocyclyl (e.g., piperidyl, tetrahydropyridyl (e.g., 1,2,3,6-tetrahydropyridyl), piperazinyl)), C 6~10aryl (e.g., phenyl) and 5-10 membered heteroaryl (e.g., 5-6 membered nitrogen-containing heteroaryl, 5-6 membered oxygen-containing heteroaryl, 5-6 membered sulfur-containing heteroaryl), wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C 1~6 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), C 1~6 Alkoxy (e.g., methoxy, ethoxy) and -C 1~6 Alkylene-NH 2 (For example, -CH 2 -NH 2 ) is optionally substituted with one or more groups selected from the group consisting of:

[0080] In some embodiments, L 6 is selected from the group consisting of 3-12 membered heterocyclyl (e.g., 3-6 membered saturated or unsaturated nitrogen-containing monoheterocyclyl (e.g., piperidinyl, tetrahydropyridyl (e.g., 1,2,3,6-tetrahydropyridyl), piperazinyl) or 3-6 membered saturated or unsaturated oxygen-containing monoheterocyclyl (e.g., tetrahydropyranyl)). In some embodiments, heterocyclyl is selected from the group consisting of C 1~6 Substituted with alkyl (eg, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl).

[0081] In some embodiments, L 6 is selected from the group consisting of 3-12 membered heterocyclyl (e.g., 3-6 membered saturated or unsaturated nitrogen-containing monoheterocyclyl (e.g., piperidinyl, tetrahydropyridyl (e.g., 1,2,3,6-tetrahydropyridyl), piperazinyl)). In some embodiments, heterocyclyl is 1~6Substituted with alkyl (eg, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl).

[0082] In some embodiments, L 7 is a covalent bond.

[0083] In some embodiments, L 8 is hydrogen, C 1~6 Alkyl, -(O-CH 2 CH 2 ) q -OC 1~6 Alkyl (e.g., -(O-CH 2 CH 2 ) q -O-methyl, -(O-CH 2 CH 2 ) q -O-ethyl) and -C 1~6 Alkylene-(O-CH 2 CH 2 ) q -OC 1~6 Alkyl (e.g., -CH 2 -(O-CH 2 CH 2 ) q -O-methyl, -CH 2 -CH 2 -(O-CH 2 CH 2 ) q -O-methyl, -CH 2 -(O-CH 2 CH 2 ) q -O-ethyl, -CH 2 -CH 2 -(O-CH 2 CH 2 ) q -O-ethyl), where q is each independently selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7; alkyl (e.g., methyl or ethyl) is selected from the group consisting of hydrogen, halogen, hydroxyl, -NH 2 , -NH-C(O)-OC 1~6 Alkyl (e.g., -NH-C(O)-O-methyl), C1~6 Alkoxy (e.g., methoxy, ethoxy), carboxyl, and -C(O)-OC 1~6 and optionally substituted with one or more groups selected from the group consisting of alkyl (e.g., -C(O)-O-methyl).

[0084] In some embodiments, L 8 is hydrogen, C 1~6 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), -CH 2 -CH 2 -O-CH 3 , -CH 2 -CH 2 -O-CH 2 CH 2 -O-CH 3 and -CH 2 -CH 2 -(O-CH 2 CH 2 ) 2 -O-CH 3 is selected from the group consisting of:

[0085] In some embodiments, L 8 is hydrogen, C 1~6 It is selected from the group consisting of alkyl (eg, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl).

[0086] In some embodiments, R 3 is hydrogen, C 1~6 Alkyl (e.g., methyl), C 1~6 3-6 membered saturated or unsaturated nitrogen-containing monoheterocyclyl (e.g., piperidyl, tetrahydropyridyl (e.g., 1,2,3,6-tetrahydropyridyl), piperazinyl) optionally substituted with alkyl (e.g., methyl); 1~6 Alkylene-C 1~6 Alkoxy (e.g., -CH 2 -CH 2 -O-CH 3), -C 1~6 Alkylene-(O-CH 2 CH 2 ) q -OC 1~6 Alkyl (e.g., -CH 2 -CH 2 -O-CH 2 CH 2 -O-CH 3 and -CH 2 -CH 2 -(O-CH 2 CH 2 ) 2 -O-CH 3 ), C 1~6 and q is independently selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7.

[0087] In some embodiments, R 3 is hydrogen, C 1~6 Alkyl (e.g., methyl), C 1~6 It is selected from the group consisting of 3-6 membered saturated or unsaturated nitrogen-containing monoheterocyclyl (eg, piperidyl, tetrahydropyridyl (eg, 1,2,3,6-tetrahydropyridyl), piperazinyl) optionally substituted with alkyl (eg, methyl).

[0088] In some embodiments, R 3 is hydrogen, methyl, tetrahydropyranyl, and piperidinyl, 1,2,3,6-tetrahydropyridyl and methyl-substituted piperazinyl, -CH 2 -CH 2 -O-CH 3 , -CH 2 -CH 2 -O-CH 2 CH 2 -O-CH 3 and -CH 2 -CH 2 -(O-CH 2 CH 2 ) 2 -O-CH 3is selected from the group consisting of:

[0089] In some embodiments, R 3 is selected from the group consisting of hydrogen, methyl, and piperidinyl, 1,2,3,6-tetrahydropyridyl, and piperazinyl substituted with methyl.

[0090] In some embodiments, R 4 is hydrogen, halogen, cyano, C 1~6 Alkyl, C 1~6 Alkoxy and C 3~10 In some embodiments, R 4 is hydrogen, C 1~3 Alkyl, C 1~3 Alkoxy and C 3~6 In some embodiments, R 4 is hydrogen.

[0091] In some embodiments, each R 5 is hydrogen and C 1~3 alkyl (e.g., methyl, ethyl, n-propyl, isopropyl); preferably, R 5 is hydrogen.

[0092] In some embodiments, each R 6 is hydrogen and C 1~3 is independently selected from the group consisting of alkyl (eg, methyl, ethyl, n-propyl, isopropyl).

[0093] In some embodiments, n and q are each independently selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25.

[0094] The present invention encompasses compounds of formula (I) obtained by any combination of the preferred groups described above. In some embodiments, the compound of the present invention has formula (II-A), formula (II-B), formula (II-C), formula (II-D), formula (II-E), or formula (II-F):

[0095] [ka] (In the formula, R 1 , R 2 and R 3 Each of the groups has the structure: In some embodiments, the compound of the present invention has formula (III-A) or formula (III-B):

[0096] [ka] (In the formula, R 1 , R 3 , L 3 and L 4 Each of the groups has the structure: In some embodiments, the compound of the present invention has formula (IV-A) or formula (IV-B):

[0097] [ka] (In the formula, R 1 , L 3 , L 4 , L 7 and L 8 Each of the groups has the structure: In some embodiments, the compound of the present invention has formula (VA) or formula (VB):

[0098] [ka] (In the formula, L 3 , L 4 , L 7 and L 8Each of the groups has the structure: In some embodiments, the compound of the present invention is

[0099] [ka] TIFF2025501049000010.tif168149 TIFF2025501049000011.tif173149 TIFF2025501049000012.tif174149 TIFF2025501049000013.tif148149 TIFF2025501049000014.tif191149 is selected from the group consisting of: The present invention also relates to a drug-linker: D-L' where D is a fragment of the compound described above; and L' is a linker), or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof. In some embodiments, L′ is -L 9 -L 10 -L 11 (In the formula, L 9 is non-existent, or

[0100] [ka] Selected from; L 10 is absent or -(CH 2 CH 2 O) s CH 2 CH 2 NH-, -(OCH 2 CH 2 ) s NH- and -C(O)-(CH2 CH 2 O) s CH 2 CH 2 NH-; s is selected from the group consisting of integers from 1 to 10; L 11 teeth,

[0101] [ka] and Z 1 is a chemical bond and

[0102] [ka] selected from the group consisting of; Z 2 is a chemical bond and C 1~20 alkylene; Z 3 is a chemical bond, C 2~6 Alkenylene and C 2~6 alkynylene; A is,

[0103] [ka] selected from the group consisting of; LG is a leaving group for nucleophilic substitution reactions, and is a halogen, nitro, benzenesulfonate, p-toluenesulfonate, trifluoromethanesulfonate, -S(O) 2 -R 7 and -S(O)-R 7 selected from the group consisting of; R 7 is C 1~6 Alkyl and C 1~6 haloalkyl). In some embodiments, L' is

[0104] [ka] TIFF2025501049000020.tif196149 is selected from. The present invention also relates to a compound having the structure:

[0105] [ka] TIFF2025501049000022.tif204149 TIFF2025501049000023.tif178149 TIFF2025501049000024.tif159149 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, comprising: The present invention also relates to a compound of formula (ISAC-I):

[0106] [ka] (In the formula, D is a fragment of a compound described in any one of the preceding items; L is a fragment of a linker L', where L' is as defined in any of the preceding items; The Ab is an antibody capable of targeting a target antigen, optionally the antibody is modified (e.g., modified with a cross-linking agent); z is selected from the group consisting of 1 to 10; Preferably, the immunostimulatory antibody conjugate has a DAR value of, for example, 1 to 10, such as: 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 2 to 3, 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 2 to 10, 3 to 4, 3 to 5, 3 to 6, 3 to 7, 3 to 8, 3 to 9, 3 to 10, 4 to 5, 4 to 6, 4 to 7, 4 to 8, 4 to 9, 4 to 10, 5 to 6, 5 to 7, 5 to 8, 5 to 9, 5 to 10, 6 to 7, 6 to 8, 6 to 9, 6 to 10, 7 to 8, 7 to 9, 7 to 10, 8 to 9, 8 to 10, or 9 to 10, preferably 1 to 8, such as 1.0 to 1.5, 1.0~2.0, 1.0~2.5, 1.0~3.0, 1.0~3.5, 1.0~4.0, 1.0~4.5, 1.0~5.0, 1.0~5.5, 1.0~6.0, 1.0~6.5, 1.0~7.0, 1.0~7.5, 1.0~8.0, 1.5~2.0, 1.5~2.5, 1 .5~3.0, 1.5~3.5, 1.5~4.0, 1.5~4.5, 1.5~5.0, 1.5~5.5, 1.5~6.0, 1.5~6.5, 1.5~7.0, 1.5~7.5, 1.5~8.0, 2.0~2.5, 2.0~3.0, 2.0~3.5, 2.0~4.0, 2.0~4. 5, 2.0~5.0, 2.0~5.5, 2.0~6.0, 2.0~6.5, 2.0~7.0, 2.0~7.5, 2.0~8.0, 2.5~3.0, 2.5~3.5, 2.5~4.0, 2.5~4.5, 2.5~5.0, 2.5~5.5, 2.5~6.0, 2.5~6.5, 2.5 ~7.0, 2.5~7.5, 2.5~8.0, 3.0~3.5, 3.0~4.0, 3.0~4.5, 3.0~5.0, 3.0~5.5, 3.0~6.0, 3.0~6.5, 3.0~7.0, 3.0~7.5, 3.0~8.0, 3.5~4.0, 3.5~4.5, 3.5~5.0, 3.5~5.5, 3.5~6.0, 3.5~6.5, 3.5~7.0, 3.5~7.5, 3.5~8.0, 4.0~4.5, 4.0~5.0, 4.0~5.5, 4.0~6.0, 4.0~6.5, 4.0~7.0, 4.0~7.5, 4.0~8.0, 4.5~5.0, 4.5~5 .5, 4.5~6.0, 4.5~6.5, 4.5~7.0, 4.5~7.5, 4.5~8.0, 5.0~5.5, 5.0~6.0, 5.0~6.5, 5.0~7.0, 5.0~7.5, 5.0~8.0, 5.5~6.0, 5.5~6.5, 5.5~7.0, 5.5~7.5, 5.5-8.0, 6.0-6.5, 6.0-7.0, 6.0-7.5, 6.0-8.0, 6.5-7.0, 6.5-7.5, 6.5-8.0, 7.0-7.5, 7.0-8.0 or 7.5-8.0), or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof.

[0107] In some embodiments, L is a structure formed by addition or nucleophilic substitution reaction of L' as described above. In some particular embodiments, L is the corresponding linker fragment having a succinimide moiety generated by addition reaction of a maleimide of L', or L is a linker formed by removal of the leaving group (e.g., methylsulfonyl) of L' via a nucleophilic substitution reaction.

[0108] In some embodiments, the crosslinker contains two identical or different reactive groups (e.g., carboxyl, carbonyl, ester, hydroxyl, thiol, amino, etc.). In some embodiments, the reactive groups in the crosslinker are free or protected. In some embodiments, the antibody reacts with one reactive group in the crosslinker to obtain a crosslinked modified antibody. In some embodiments, in the crosslinked modified antibody, the other reactive group of the crosslinker moiety that does not react with the antibody is in protected or deprotected form. In some embodiments, one of the two reactive groups of the crosslinker is linked to the antibody and the other is linked to L. In some embodiments, the crosslinker is the crosslinker SATA. In some embodiments, formula (ISAC-I) has the structure:

[0109] [ka] has. The present invention also relates to a compound of formula (ISAC-I):

[0110] [ka] (In the formula, D is selected from the group consisting of fragments of the compound of formula (I); L is a linker; The Ab is an antibody capable of targeting a target antigen, such as an antibody that targets a tumor cell; optionally, the antibody is modified (e.g., modified with a cross-linking agent); such as a trastuzumab monoclonal antibody or a trastuzumab monoclonal antibody modified with the cross-linking agent SATA; and z is selected from the group consisting of 1 to 10), or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

[0111] How to prepare Another object of the present invention is to provide a method for preparing the compounds of the present invention. For example, the present invention provides a method for preparing a compound of formula (II-A), comprising: 2 is not hydrogen, but R 3 is not hydrogen, the following steps:

[0112] [ka] Including, X represents a halogen atom, including chlorine, bromine and iodine. R 3 When R is methyl, II-A-9 is trimethylboroxane. 3 is not methyl, II-A-9 is

[0113] [ka] It is.

[0114] R 1 , R 2 and R 3 is as defined above, and R2 Also 3 nor hydrogen; (1) reacting compound II-A-1 and compound II-A-2 to obtain compound II-A-3; The reaction is preferably carried out in a suitable organic solvent. The organic solvent can be selected from the group consisting of N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, toluene, tetrahydrofuran and any combination thereof, preferably N,N-dimethylformamide. The reaction is preferably carried out in the presence of a suitable organic or inorganic base, which can be selected from the group consisting of N,N-diisopropylethylamine, triethylamine, 4-dimethylaminopyridine, potassium carbonate, sodium carbonate, preferably N,N-diisopropylethylamine. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 1 to 12 hours.

[0115] (2) subjecting compound II-A-3 to a reduction reaction to obtain compound II-A-4; The reaction is preferably carried out under a reduction system of sodium dithionite, a palladium metal catalyst or a platinum metal catalyst, and hydrogen. The palladium metal catalyst or the platinum metal catalyst can be selected from the group consisting of palladium on carbon, palladium hydroxide, platinum dioxide, preferably palladium on carbon. The reaction is preferably carried out in a suitable solvent, which can be selected from the group consisting of methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, water, and any combination thereof. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 2 to 16 hours.

[0116] (3) reacting compound II-A-4 with compound II-A-5 to obtain compound II-A-6; The reaction is preferably carried out in a suitable organic solvent. The organic solvent can be selected from the group consisting of N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, dichloromethane, tetrahydrofuran and any combination thereof, preferably tetrahydrofuran. The reaction is preferably carried out in the presence of a suitable organic or inorganic base, which can be selected from the group consisting of N,N-diisopropylethylamine, triethylamine, 4-dimethylaminopyridine, potassium carbonate, sodium carbonate, preferably N,N-diisopropylethylamine. The reaction is preferably carried out at a suitable temperature, preferably -10 to 60°C. The reaction is preferably carried out for a suitable time, for example, 2 to 12 hours.

[0117] (4) subjecting compound II-A-6 to a ring-closure reaction to obtain compound II-A-7; The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of toluene, xylene, mesitylene, N,N-dimethylformamide, N-methylpyrrolidone, and any combination thereof, preferably toluene. The reaction is preferably carried out with the participation of o-chlorobenzoic acid. The reaction is preferably carried out at a suitable temperature, preferably 50 to 150° C. The reaction is preferably carried out for a suitable time, for example, 2 to 12 hours.

[0118] (5) subjecting compound II-A-7 to a halogenation reaction to obtain compound II-A-8; The reaction is preferably carried out in a suitable organic solvent. The organic solvent can be selected from the group consisting of N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, formic acid, glacial acetic acid, and any combination thereof, preferably a combination of N,N-dimethylformamide and glacial acetic acid. The reaction is preferably carried out in the presence of a suitable halogenating agent, which can be selected from the group consisting of N-chlorosuccinimide, N-bromosuccinimide, dibromohydantoin, bromine, N-iodosuccinimide, and any combination thereof, preferably N-bromosuccinimide. The reaction is preferably carried out at a suitable temperature, preferably -10 to 60°C. The reaction is preferably carried out for a suitable time, for example, 2 to 36 hours.

[0119] (6) subjecting compound II-A-8 and compound II-A-9 to a coupling reaction to obtain compound II-A-10; The coupling reaction is preferably carried out in the presence of a metal catalyst and a base. Preferably, the metal catalyst is a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, 1,1'-bisdiphenylphosphinoferrocenepalladium dichloride, tetrakis(triphenylphosphine)palladium, palladium acetate, preferably 1,1'-bisdiphenylphosphinoferrocenepalladium dichloride. The base is an organic base or an inorganic base, such as N,N-diisopropylethylamine, triethylamine, sodium tert-butoxide, potassium carbonate, cesium carbonate, sodium carbonate, preferably potassium carbonate. The reaction is preferably carried out in a suitable solvent, which may be selected from the group consisting of N,N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, 1,4-dioxane, water, and any combination thereof, preferably a combination of 1,4-dioxane and water. The reaction is preferably carried out at a suitable temperature, preferably 60 to 150°C. The reaction is preferably carried out for an appropriate time, for example, 1 to 12 hours.

[0120] (7) reacting compound II-A-10 to obtain compound II-A-11; The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of N,N-dimethylformamide, dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, and any combination thereof, preferably dichloromethane. The reaction is preferably carried out in the presence of a suitable oxidizing agent, which may be selected from the group consisting of m-chloroperoxybenzoic acid, hydrogen peroxide, tert-butyl hydroperoxide, and any combination thereof, preferably m-chloroperoxybenzoic acid. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 1 to 12 hours.

[0121] (8) reacting compound II-A-11 to obtain compound II-A; The reaction is preferably carried out in a suitable solvent. The solvent can be selected from the group consisting of N,N-dimethylformamide, dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, aqueous ammonia, and any combination thereof, and is preferably a combination of dichloromethane and aqueous ammonia. The reaction is preferably carried out in the presence of p-toluenesulfonyl chloride. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 2 to 24 hours. In particular, the present invention also provides a process for preparing a compound of formula (II-A'), comprising the steps of: 3 is hydrogen, and R 2 is not hydrogen, so next step:

[0122] [ka] (In the formula, R 1 and R 2 is as defined above, and R 2 is not hydrogen); (1) reacting compound II-A-7 to obtain compound II-A-12; The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of N,N-dimethylformamide, dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, and any combination thereof, preferably dichloromethane. The reaction is preferably carried out in the presence of a suitable oxidizing agent, which may be selected from the group consisting of m-chloroperoxybenzoic acid, hydrogen peroxide, tert-butyl hydroperoxide, and any combination thereof, preferably m-chloroperoxybenzoic acid. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 1 to 12 hours.

[0123] (2) reacting compound II-A-12 to obtain compound II-A'; The reaction is preferably carried out in a suitable solvent. The solvent can be selected from the group consisting of N,N-dimethylformamide, dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, aqueous ammonia, and any combination thereof, and is preferably a combination of dichloromethane and aqueous ammonia. The reaction is preferably carried out in the presence of p-toluenesulfonyl chloride. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 2 to 24 hours. In particular, the present invention also provides a method for preparing a compound of formula (II-A″), comprising the steps of: 2 is hydrogen, and R 3 is not hydrogen, so next step:

[0124] [ka] where X represents a halogen atom, including chlorine, bromine and iodine. R 3 When R is methyl, II-A-9 is trimethylboroxine.3 is not methyl, IA-9 is

[0125] [ka] It is. R 1 and R 3 is as defined above, and R 3 is not hydrogen; (1) subjecting compound II-A-1 to a substitution reaction with ammonia in methanol to obtain compound II-A-2'; The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of methanol, ethanol, isopropanol, 1,4-dioxane, tetrahydrofuran, and any combination thereof, and is preferably methanol. The reaction is preferably carried out at a suitable temperature, preferably 0 to 100° C. The reaction is preferably carried out for a suitable time, for example, 1 to 12 hours.

[0126] (2) subjecting compound II-A-2' to a reduction reaction to obtain compound II-A-3'; The reaction is preferably carried out under a reduction system of sodium dithionite, a palladium metal catalyst or a platinum metal catalyst, and hydrogen. The palladium metal catalyst or the platinum metal catalyst can be selected from the group consisting of palladium on carbon, palladium hydroxide, platinum dioxide, preferably palladium on carbon. The reaction is preferably carried out in a suitable solvent, which can be selected from the group consisting of methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, water, and any combination thereof. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 2 to 16 hours.

[0127] (3) reacting compound II-A-3' with compound II-A-5 to obtain compound II-A-4'; The reaction is preferably carried out in a suitable organic solvent. The organic solvent can be selected from the group consisting of N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, dichloromethane, tetrahydrofuran and any combination thereof, preferably tetrahydrofuran. The reaction is preferably carried out in the presence of a suitable organic or inorganic base, which can be selected from the group consisting of N,N-diisopropylethylamine, triethylamine, 4-dimethylaminopyridine, potassium carbonate, sodium carbonate, preferably N,N-diisopropylethylamine. The reaction is preferably carried out at a suitable temperature, preferably -10 to 60°C. The reaction is preferably carried out for a suitable time, for example, 2 to 12 hours.

[0128] (4) subjecting compound II-A-4' to a ring-closing reaction to obtain compound II-A-5'; The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of methanol, ethanol, isopropanol, 1,4-dioxane, tetrahydrofuran, and any combination thereof, preferably ethanol. The reaction is preferably carried out in the presence of a suitable organic or inorganic base, which may be selected from the group consisting of N,N-diisopropylethylamine, triethylamine, sodium hydroxide, potassium hydroxide, potassium carbonate, sodium carbonate, preferably sodium hydroxide. The reaction is preferably carried out at a suitable temperature, preferably 0 to 100°C. The reaction is preferably carried out for a suitable time, for example, 2 to 12 hours.

[0129] (5) subjecting compound II-A-5' to a halogenation reaction to obtain compound II-A-6'; The reaction is preferably carried out in a suitable organic solvent. The organic solvent can be selected from the group consisting of N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, formic acid, glacial acetic acid, and any combination thereof, preferably a combination of N,N-dimethylformamide and glacial acetic acid. The reaction is preferably carried out in the presence of a suitable halogenating agent, which can be selected from the group consisting of N-chlorosuccinimide, N-bromosuccinimide, dibromohydantoin, bromine, N-iodosuccinimide, and any combination thereof, preferably N-bromosuccinimide. The reaction is preferably carried out at a suitable temperature, preferably -10 to 60°C. The reaction is preferably carried out for a suitable time, for example, 2 to 36 hours.

[0130] (6) reacting compound II-A-6' with di-tert-butyl dicarbonate to obtain a mixture of compound II-A-7-1' and compound II-A-7-2'; The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of N,N-dimethylformamide, dichloromethane, tetrahydrofuran and any combination thereof, preferably tetrahydrofuran. The reaction is preferably carried out in the presence of a suitable organic or inorganic base, which may be selected from the group consisting of N,N-diisopropylethylamine, triethylamine, 4-dimethylaminopyridine, potassium carbonate, sodium carbonate, preferably a mixture of triethylamine and 4-dimethylaminopyridine. The reaction is preferably carried out at a suitable temperature, preferably -10 to 60°C. The reaction is preferably carried out for a suitable time, for example, 2 to 24 hours.

[0131] (7) reacting the mixture of compound II-A-7-1′ and compound II-A-7-2′ to obtain a mixture of compound II-A-8-1′ and compound II-A-8-2′; The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of N,N-dimethylformamide, dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, and any combination thereof, preferably dichloromethane. The reaction is preferably carried out in the presence of a suitable oxidizing agent, which may be selected from the group consisting of m-chloroperoxybenzoic acid, hydrogen peroxide, tert-butyl hydroperoxide, and any combination thereof, preferably m-chloroperoxybenzoic acid. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 1 to 12 hours.

[0132] (8) reacting the mixture of compound II-A-8-1′ and compound II-A-8-2′ with tert-butylamine to obtain a mixture of compound II-A-9-1′ and compound II-A-9-2′; The reaction is preferably carried out in a suitable solvent. The solvent may be selected from the group consisting of N,N-dimethylformamide, dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, and any combination thereof, and is preferably dichloromethane. The reaction is preferably carried out in the presence of p-toluenesulfonyl chloride. The reaction is preferably carried out at a suitable temperature, preferably from -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, from 2 to 24 hours.

[0133] (9) subjecting the mixture of compound II-A-9-1′ and compound II-A-9-2′ to a coupling reaction with compound II-A-9 to obtain compound II-A-10′; The coupling reaction is preferably carried out in the presence of a metal catalyst and a base. Preferably, the metal catalyst is a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, 1,1'-bisdiphenylphosphinoferrocenepalladium dichloride, tetrakis(triphenylphosphine)palladium, palladium acetate, preferably 1,1'-bisdiphenylphosphinoferrocenepalladium dichloride. The base is an organic base or an inorganic base, such as N,N-diisopropylethylamine, triethylamine, sodium tert-butoxide, potassium carbonate, cesium carbonate, sodium carbonate, preferably potassium carbonate. The reaction is preferably carried out in a suitable solvent, which may be selected from the group consisting of N,N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, 1,4-dioxane, water, and any combination thereof, preferably a combination of 1,4-dioxane and water. The reaction is preferably carried out at a suitable temperature, preferably 60 to 150°C. The reaction is preferably carried out for an appropriate time, for example, 1 to 12 hours.

[0134] (10) reacting compound II-A-10′ to obtain compound II-A″; The organic solvent can be selected from the group consisting of methanol, ethanol, isopropanol, 1,4-dioxane, and any combination thereof, and is preferably methanol. The reaction is carried out in the presence of a suitable inorganic acid selected from the group consisting of sulfuric acid, hydrochloric acid, hydrobromic acid, and any combination thereof, preferably hydrochloric acid. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 1 to 12 hours. In particular, the present invention provides a process for preparing a compound of formula (II-A), 2 is not hydrogen, but R 3 is not hydrogen, the following steps:

[0135] [ka] (In the formula, R 1 , R 2 and R 3 is as defined above, and R 2 and R 3 is not hydrogen); (1) reacting compound II-A-11 with tert-butylamine to obtain compound II-A-13; The reaction is preferably carried out in a suitable solvent. The solvent may be selected from the group consisting of N,N-dimethylformamide, dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, and any combination thereof, and is preferably dichloromethane. The reaction is preferably carried out in the presence of p-toluenesulfonyl chloride. The reaction is preferably carried out at a suitable temperature, preferably from -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, from 2 to 24 hours.

[0136] (2) reacting compound II-A-13 to obtain compound II-A; The reaction is preferably carried out in a suitable organic solvent. The organic solvent can be selected from the group consisting of methanol, ethanol, isopropanol, 1,4-dioxane, and any combination thereof, and is preferably methanol. The reaction is carried out in the presence of a suitable inorganic acid selected from the group consisting of sulfuric acid, hydrochloric acid, hydrobromic acid, and any combination thereof, preferably hydrochloric acid. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 1 to 12 hours. In particular, the present invention also provides a process for preparing a compound of formula (II-A), comprising the steps of: 2 is not hydrogen, R 3 is not hydrogen, so next step:

[0137] [ka] where X represents a halogen atom, including chlorine, bromine and iodine. R 3 When R is methyl, II-A-9 is trimethylboroxine. 3 is not methyl, II-A-9 is

[0138] [ka] It is. R 1 , R 2 and R 3 is as defined above, and R 2 Also 3 nor hydrogen; (1) reacting compound II-A-8 to obtain compound II-A-14; The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of N,N-dimethylformamide, dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, and any combination thereof, preferably dichloromethane. The reaction is preferably carried out in the presence of a suitable oxidizing agent, which may be selected from the group consisting of m-chloroperoxybenzoic acid, hydrogen peroxide, tert-butyl hydroperoxide, and any combination thereof, preferably m-chloroperoxybenzoic acid. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 1 to 12 hours.

[0139] (2) reacting compound II-A-14 with tert-butylamine to obtain compound II-A-15; The reaction is preferably carried out in a suitable solvent. The solvent may be selected from the group consisting of N,N-dimethylformamide, dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, and any combination thereof, and is preferably dichloromethane. The reaction is preferably carried out in the presence of p-toluenesulfonyl chloride. The reaction is preferably carried out at a suitable temperature, preferably from -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, from 2 to 24 hours.

[0140] (3) subjecting compound II-A-15 and compound II-A-9 to a coupling reaction to obtain compound II-A-13; The coupling reaction is preferably carried out in the presence of a metal catalyst and a base. Preferably, the metal catalyst is a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, 1,1'-bisdiphenylphosphinoferrocenepalladium dichloride, tetrakis(triphenylphosphine)palladium, palladium acetate, preferably 1,1'-bisdiphenylphosphinoferrocenepalladium dichloride. The base is an organic base or an inorganic base, such as N,N-diisopropylethylamine, triethylamine, sodium tert-butoxide, potassium carbonate, cesium carbonate, sodium carbonate, preferably potassium carbonate. The reaction is preferably carried out in a suitable solvent, which may be selected from the group consisting of N,N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, 1,4-dioxane, water, and any combination thereof, preferably a combination of 1,4-dioxane and water. The reaction is preferably carried out at a suitable temperature, preferably 60 to 150°C. The reaction is preferably carried out for an appropriate time, for example, 1 to 12 hours.

[0141] (4) reacting compound II-A-13 to obtain compound II-A; The reaction is preferably carried out in a suitable organic solvent. The organic solvent can be selected from the group consisting of methanol, ethanol, isopropanol, 1,4-dioxane, and any combination thereof, and is preferably methanol. The reaction is carried out in the presence of a suitable inorganic acid selected from the group consisting of sulfuric acid, hydrochloric acid, hydrobromic acid, and any combination thereof, preferably hydrochloric acid. The reaction is preferably carried out at a suitable temperature, preferably -10 to 80°C. The reaction is preferably carried out for a suitable time, for example, 1 to 12 hours.

[0142] The detailed conditions of the above reaction steps are well known in the art, and the present invention is not particularly limited thereto. According to the teachings of the present invention combined with the general knowledge in the art, those skilled in the art can select and replace each substituent in the general formula to prepare different compounds, and these selections and replacements are all within the protection scope of the present invention.

[0143] Pharmaceutical Compositions and Kits Another object of the present invention is to provide pharmaceutical compositions comprising a prophylactically or therapeutically effective amount of a compound of the present invention or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, and one or more pharma- ceutically acceptable carriers.

[0144] Another object of the present invention is to provide a kit comprising a compound of the present invention or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, or a pharmaceutical composition of the present invention. Optionally, the kit further comprises instructions for use.

[0145] As used herein, a "pharmaceutical acceptable carrier" refers to a diluent, adjuvant, excipient, or vehicle with which a therapeutic is administered that is suitable for contact with tissues obtained from humans and / or other animals without undue toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0146] Pharmaceutically acceptable carriers that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, sterile liquids. The pharmaceutical compositions can be in the form of, for example, a solid, semi-solid, liquid or gaseous preparation.

[0147] The pharmaceutical compositions of the present invention may act systemically and / or locally. For this purpose, the pharmaceutical compositions may be administered by a suitable route, for example by injection or transdermal administration; or by oral administration or inhalation administration.

[0148] The content or amount of the compound of the present invention in the pharmaceutical composition may be about 0.001 mg to about 1000 mg, suitably 0.01 to 800 mg, preferably 0.05 to 500 mg.

[0149] In some embodiments, the present invention provides methods for preparing a pharmaceutical composition of the present invention, the methods comprising combining a compound of the present invention or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof with one or more pharma- ceutically acceptable carriers.

[0150] Treatment Methods and Uses The compounds of the present invention or their pharma- ceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs, or pharmaceutical compositions of the present invention have potent agonistic activity against the target TLR7 and / or TLR8.

[0151] Another object of the present invention is to provide a use of the compounds of the present invention in the manufacture of an immunostimulatory antibody conjugate, in some embodiments, the immunostimulatory antibody conjugate comprises a small molecule ligand capable of interacting with TLR7 and / or TLR8.

[0152] Another object of the present invention is to provide the use of a compound of the present invention or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, or an immunostimulatory antibody conjugate of the present invention, or a pharmaceutical composition of the present invention, in the manufacture of a medicament for the prevention and / or treatment of a disease mediated by TLR7 and / or TLR8.

[0153] Another object of the present invention is to provide a method for preventing and / or treating a disease mediated by TLR7 and / or TLR8, comprising the step of administering to a subject in need thereof a prophylactically or therapeutically effective amount of a compound of the present invention or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug, or an immunostimulatory antibody conjugate of the present invention, or a pharmaceutical composition of the present invention.

[0154] In some embodiments, the disease mediated by TLR7 and / or TLR8 is preferably a tumor.

[0155] As used herein, the term "effective amount" refers to an amount sufficient to achieve a desired prophylactic or therapeutic effect, e.g., an amount that achieves the alleviation of one or more symptoms associated with the disease being treated.

[0156] Dosage regimens can be adjusted to provide the optimum desired response. For example, a single bolus can be administered, several divided doses can be administered over time, or the dosage can be proportionally increased or decreased as indicated by the exigencies of the therapeutic situation. It should be noted that dosages vary with the type and severity of the condition to be alleviated, and can include single or multiple administrations. It should further be understood that for any given individual, the specific dosage regimen will be adjusted over time according to the individual's needs and the professional judgment of the person administering or supervising the administration of the compounds of the present invention.

[0157] The amount of a compound of this invention administered will depend on the individual being treated, the severity of the disease or condition, the rate of administration, the compound's therapeutic effect, and the judgment of the prescribing physician. In some cases, dosages up to the lower end of the range may be sufficient, while in other cases, larger doses may be used without causing any adverse side effects, provided that the larger dose is initially divided into several smaller doses to be administered throughout the day.

[0158] As used herein, unless otherwise indicated, the term "treatment" refers to reversing, alleviating, or ameliorating the progression of the disease or condition to which such term applies, or one or more symptoms of such disease or condition.

[0159] The term "prevention" refers to inhibiting and delaying the onset of a disease, including not only prevention before onset, but also prevention of recurrence of the disease after treatment.

[0160] As used herein, the term "subject" includes humans or non-human animals. Exemplary human subjects include human subjects (referred to as patients) suffering from a disease (e.g., a disease described herein) or normal subjects. "Non-human animals" in the present invention include 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.).

[0161] The accompanying drawings described in this specification are used to provide a further understanding of the present invention and constitute a part of this application. The schematic examples of the present invention and their description are used to explain the present invention and are not to be construed as unduly limiting the present invention. [Brief description of the drawings]

[0162] [Figure 1] FIG. 1 shows the curves of changes in tumor volume over time in the groups of Experimental Example 4. Detailed Model for Implementing the Invention

[0163] In order to make the objectives and technical solutions of the present invention clearer, the embodiments of the present invention will be described in detail below in combination with examples. However, those skilled in the art will understand that the following examples are only used to illustrate the present invention, and should not be considered as any limitation of the scope of the present invention. In these examples, those without specific conditions were carried out according to conventional conditions or conditions suggested by the manufacturers. Reagents or equipment not indicated by the manufacturers are all commercially available conventional products.

[0164] The structure of the compound was confirmed by nuclear magnetic resonance ( 1 The molecular weight was determined by 1 H NMR or mass spectrometry (MS). 1 H NMR was determined using a JEOL Eclipse 400 NMR instrument. The solvent used was deuterated methanol (CD 3 OD), deuterated chloroform (CDCl 3 ) or hexadeuteriodimethylsulfoxide (DMSO-d 6 ), the internal standard was tetramethylsilane (TMS), and chemical shifts (δ) were in parts per million (ppm).

[0165] MS was measured using an Agilent (ESI) mass spectrometer: manufacturer: Agilent, model: Agilent 6120B. High performance liquid chromatography preparation method: Instrument model: Agilent 1260; Chromatography column: Waters SunFire Prep C18 OBD (19 mm × 150 mm × 5.0 μm); Chromatography column temperature: 25 °C; Flow rate: 20.0 mL / min; Detection wavelength: 214 nm; Elution gradient: (0 min: A 10%, B 90%; 16.0 min: A 90%, B 10%); Mobile phase A: acetonitrile; Mobile phase B: 0.05% formic acid in water. Aluminum plates (20×20 cm) from Merck were used as silica gel plates for thin layer chromatography (TLC), and GF 254 (1 mm) from Yantai was used for TLC separation and purification.

[0166] The reaction was monitored by thin layer chromatography (TLC) or LC-MS; the developing solvent systems included dichloromethane / methanol, n-hexane / ethyl acetate, and petroleum ether / ethyl acetate, and the volume ratio of the solvents was adjusted according to the polarity of the compounds or by adding triethylamine, etc.

[0167] For microwave reactions, a Biotage Initiator+ (400W, RT approximately 300° C.) microwave reactor was used.

[0168] Column chromatography generally uses 200-300 mesh silica gel as a carrier. The eluent system includes dichloromethane / methanol system and petroleum ether / ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and may also be adjusted by adding a small amount of triethylamine. Unless otherwise specified in these examples, reaction temperatures were room temperature (20°C to 35°C); Reagents used in the present invention were purchased from companies such as Acros Organics, Aldrich Chemical Company, and Topbiochem Ltd.

[0169] In the general synthesis methods and synthesis examples, as well as intermediate synthesis examples, the meanings of the abbreviations are as follows:

[0170] [Table 1]

[0171] I. Compound examples Example 1: Preparation of 1-(4-aminobutyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 1)

[0172] [ka]

[0173] Step 1: Preparation of tert-butyl (4-((6-nitrothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate 7-Chloro-6-nitrothieno[3,2-b]pyridine (10.0 g, 46.59 mmol) and N-tert-butoxycarbonyl-1,4-butanediamine (10.53 g, 55.91 mmol) were dissolved in N,N-dimethylformamide (150 mL), cooled in an ice-water bath, N,N-diisopropylethylamine (12.04 g, 93.18 mmol) was added to the reaction solution at 0° C., slowly warmed to room temperature, and stirred for 6 hours. The reaction solution was poured into water (1500 mL), extracted three times with ethyl acetate (450 mL), the organic phase was combined, washed three times with saturated saline (150 mL), the organic phase was dehydrated over anhydrous sodium sulfate, and concentrated to obtain the title compound (16.61 g, yield: 97.3%). MS m / z (ESI): 367.1 [M+H] + .

[0174] Step 2: Preparation of tert-butyl (4-((6-aminothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate tert-Butyl (4-((6-nitrothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate (16.61 g, 49.37 mmol) was dissolved in a mixture of tetrahydrofuran (100 mL) and methanol (100 mL), added with 10% palladium on carbon (2.0 g, 11.97 mmol), exchanged with hydrogen three times, the reaction was stirred at room temperature for 12 hours, subjected to suction filtration, the filter cake was washed with methanol (30 mL), and the filtrate was concentrated to give the title compound (14.47 g, yield: 87.1%). MS m / z (ESI): 337.1 [M+H] + .

[0175] Step 3: Preparation of tert-butyl (4-((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate tert-Butyl (4-((6-aminothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate (14.47 g, 43.01 mmol) and N,N-diisopropylethylamine (11.12 g, 86.02 mmol) were dissolved in tetrahydrofuran (250 mL), cooled in an ice-water bath, and n-valeryl chloride (7.78 g, 64.52 mmol) was slowly added dropwise at 0° C., then slowly warmed to room temperature and stirred for 4 hours. Water (100 mL) was added to the reaction system to quench the reaction, and the system was concentrated to remove as much THF as possible. The resulting residue was added with water (300 mL) and extracted three times with ethyl acetate (300 mL). The organic phases were combined and washed three times with saturated brine (150 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the title compound (16.72 g, yield: 92.4%). MS m / z (ESI): 421.0 [M+H] + .

[0176] Step 4: Preparation of tert-butyl (4-(2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate tert-Butyl (4-((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate (16.72 g, 39.76 mmol) and o-chlorobenzoic acid (3.11 g, 19.88 mmol) were added to toluene (300 mL) and heated to reflux, then water was separated and the reaction was stirred for 4 hours. After cooling to room temperature, the reaction was concentrated to remove the solvent, and the resulting residue was purified by silica gel column chromatography (eluent: 100% ethyl acetate) to give the title compound (14.76 g, yield: 92.2%). MS m / z (ESI): 403.2 [M+H] + .

[0177] Step 5: Preparation of tert-butyl (4-(7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate tert-Butyl (4-(2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate (5.41 g, 13.44 mmol) was completely dissolved in a mixture of N,N-dimethylformamide (75 mL) and glacial acetic acid (25 mL), cooled in an ice-water bath, and N-bromosuccinimide (4.78 g, 26.88 mmol) was added slowly in batches to the reaction solution at 0° C., slowly warmed to room temperature, and stirred for 24 hours. The reaction mixture was poured into water (300 mL) and extracted three times with ethyl acetate (300 mL). The organic phases were combined and washed three times with saturated aqueous sodium bicarbonate solution (150 mL) and three times with saturated saline solution (150 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate:petroleum ether = 1:1, volume ratio) to obtain the title compound (4.95 g, yield: 76.5%). MS m / z (ESI): 481.0 [M+H] + .

[0178] Step 6: Preparation of tert-butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate tert-Butyl (4-(7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate (1.0 g, 2.08 mmol), trimethylboroxine (50 wt% in THF, 2.62 g, 10.4 mmol), potassium carbonate (575.0 mg, 4.16 mmol), 1,1'-bisdiphenylphosphinoferrocenepalladium dichloride (307.3 mg, 0.42 mmol) were added sequentially to a mixture of 1,4-dioxane (10 mL) and water (1 mL), and the reaction was sparged with nitrogen for 5 min, then heated to 100 °C and stirred under microwave for 2 h. After cooling to room temperature, the reaction system was concentrated to remove the solvent, and the resulting residue was purified by silica gel column chromatography (eluent: 100% ethyl acetate) to obtain the title compound (0.65 g, yield: 75.0%). MS m / z (ESI): 417.1 [M+H]+ .

[0179] Step 7: Preparation of 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide tert-Butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate (0.65 g, 1.56 mmol) was dissolved in dichloromethane (10 mL), cooled in an ice-water bath, m-chloroperoxybenzoic acid (538.5 mg, 3.12 mmol) was added slowly in batches to the reaction solution at 0° C., slowly warmed to room temperature, and stirred for 4 hours. The reaction solution was poured into water (100 mL), extracted three times with dichloromethane (60 mL), the organic phases were combined, washed twice with saturated aqueous sodium bicarbonate solution (30 mL), washed twice with saturated aqueous sodium sulfite solution (30 mL), washed three times with saturated brine (45 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated to give the title compound (640 mg, yield: 94.8%). MS m / z (ESI): 433.2 [M+H] + .

[0180] Step 8: Preparation of tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide (640 mg, 1.48 mmol) was dissolved in a mixed solvent of dichloromethane (10 mL) and aqueous ammonia (5 mL), cooled in an ice-water bath, and p-toluenesulfonyl chloride (564.1 mg, 2.96 mmol) was slowly added in batches to the reaction solution at 0° C., slowly warmed to room temperature, and stirred for 12 hours. The reaction mixture was poured into water (100 mL) and extracted three times with dichloromethane (60 mL). The organic phases were combined and washed three times with saturated brine (45 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate:methanol=15:1, volume ratio) to obtain the title compound (266 mg, yield: 41.6%). MS m / z (ESI): 432.2 [M+H] + .

[0181] Step 9: Preparation of 1-(4-aminobutyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 1) tert-Butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate (266 mg, 0.62 mmol) was dissolved in a mixed solvent of methanol (10 mL) and hydrogen chloride in 1,4-dioxane (4N, 5 mL) and stirred at room temperature for 12 hours. After removing the solvent by concentration, a mixed solvent of dichloromethane and methanol (volume ratio 4:1, 10 mL) and saturated aqueous sodium bicarbonate solution (2 mL) were added to the residue in turn, stirred at room temperature for 5 minutes, and then the solvent was removed by concentration. The obtained residue was purified by preparative high performance liquid chromatography to obtain the title compound (130 mg, yield: 63.3%). MS m / z (ESI): 332.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6): δ 8.34 (s, 2H), 6.98 (d, J = 1.2 Hz, 1H), 6.06 (s, 2H),4.24 (t, J = 8.0 Hz, 2H), 2.86 (t, J = 8.0 Hz, 2H), 2.78 (t, J = 8.0 Hz, 2H),2.56 (d, J = 1.2 Hz, 3H), 1.83-1.75 (m, 4H), 1.62-1.58 (m, 2H), 1.47-1.40 (m, 2H),0.96 (t, J = 8.0 Hz, 3H).

[0182] Example 2: Preparation of 1-(4-aminobutyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 2)

[0183] [ka]

[0184] Step 1: Preparation of 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide

[0185] The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 7 was replaced with tert-butyl (4-(2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate to obtain the title compound (210 mg, yield: 93.2%). MS m / z (ESI): 419.0 [M+H] + .

[0186] Step 2: Preparation of tert-butyl (4-(4-amino-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate The synthetic route of Example 1 was applied, and the raw material 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide in step 8 was replaced with 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide to obtain the title compound (152 mg, yield: 45.4%). MS m / z (ESI): 418.1 [M+H] + .

[0187] Step 3: Preparation of 1-(4-aminobutyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 2) The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl (4-(4-amino-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate to obtain the title compound (62 mg, yield: 52.5%). MS m / z (ESI): 318.0 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 8.32 (s, 2H), 7.74-7.73 (m, 1H), 7.27-7.25 (m, 1H),6.14 (s, 2H), 4.30 (t, J = 8.0 Hz, 2H), 2.91-2.87 (m, 2H), 2.79-2.76 (m, 2H),1.85-1.76 (m, 4H), 1.63-1.58 (m, 2H), 1.50-1.41 (m, 2H), 0.96 (t, J = 8.0 Hz,3H).

[0188] Example 3: Preparation of 2-butyl-7-methyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 3)

[0189] [ka] Step 1: Preparation of benzyl 4-(((6-nitrothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate Applying the synthetic route of Example 1, the starting material N-tert-butoxycarbonyl-1,4-butanediamine in step 1 was replaced with benzyl 4-(aminomethyl)tetrahydro-1(2H)-picolinate to give the title compound (1.53 g, yield: 92.5%). MS m / z (ESI): 427.0 [M+H] + .

[0190] Step 2: Preparation of benzyl 4-(((6-aminothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate Benzyl 4-(((6-nitrothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate (2.0 g, 4.69 mmol) was completely dissolved in absolute ethanol (30 mL), added with water (5 mL), cooled in an ice-water bath, sodium dithionite (8.16 g, 46.9 mmol) was added to the reaction solution at 0° C., slowly warmed to room temperature, and stirred for 4 hours. The reaction solution was poured into water (150 mL), stirred to precipitate a solid, filtered, washed with water, and dried to obtain the title compound (1.65 g, yield: 88.7%). MS m / z (ESI): 397.1 [M+H] + .

[0191] Step 3: Preparation of benzyl 4-(((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-aminothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 3 was replaced with benzyl 4-(((6-aminothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate to give the title compound (1.47 g, yield: 90.2%). MS m / z (ESI): 481.1 [M+H] + .

[0192] Step 4: Preparation of benzyl 4-((2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 4 was replaced with benzyl 4-(((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate to give the title compound (1.18 g, yield: 91.8%). MS m / z (ESI): 463.1 [M+H] + .

[0193] Step 5: Preparation of benzyl 4-((7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 5 was replaced with benzyl 4-((2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carbamate to give the title compound (726 mg, yield: 74.3%). MS m / z (ESI): 541.1 [M+H] + .

[0194] Step 6: Preparation of benzyl 4-((2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 6 was replaced with benzyl 4-((7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to obtain the title compound (623 mg, yield: 76.7%). MS m / z (ESI): 477.1 [M+H] + .

[0195] Step 7: Preparation of 1-((1-((benzyloxy)carbonyl)piperidin)-4-yl)methyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 6 was replaced with benzyl 4-((2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to obtain the title compound (517 mg, yield: 93.2%). MS m / z (ESI): 493.2 [M+H] + .

[0196] Step 8: Preparation of benzyl 4-((4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide in step 8 was replaced with 1-((1-((benzyloxy)carbonyl)piperidin-4-yl)methyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide to obtain the title compound (287 mg, yield: 38.2%). MS m / z (ESI): 492.2 [M+H] + .

[0197] Step 9: Preparation of 2-butyl-7-methyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 3) Benzyl 4-((4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate (150 mg, 0.31 mmol) was added to trifluoroacetic acid (6 mL), heated to 50° C., and stirred for 3 hours. After removing the solvent by concentration, a mixed solvent of dichloromethane and methanol (volume ratio 4:1, 10 mL) and a saturated aqueous sodium bicarbonate solution (10 mL) were added to the residue in turn, stirred at room temperature for 5 minutes, and then the solvent was removed by concentration. The obtained residue was purified by preparative high performance liquid chromatography to obtain the title compound (22 mg, yield: 19.9%). MS m / z (ESI): 358.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6): δ 8.24 (s, 1H), 6.99-6.98 (m, 1H), 6.11 (br, 2H), 4.14(d, J = 8.0 Hz, 2H), 3.26-3.23 (m, 2H), 2.88-2.76 (m, 4H), 2.57-2.55 (m, 3H),2.19-2.12(m, 1H), 1.87-1.76 (m, 2H), 1.68-1.62(m, 2H), 1.56-1.39 (m, 4H), 0.96(t, J = 8.0 Hz, 3H).

[0198] Example 4: Preparation of 1-(4-aminobutyl)-2-(2-methoxyethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 4)

[0199] [ka] Step 1: Preparation of tert-butyl (4-((6-(3-methoxypropionamido)thieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate The synthetic route of Example 1 was applied, and the raw material n-pentanoyl chloride in step 3 was replaced with 3-methoxypropionyl chloride to give the title compound (847 mg, yield: 93.2%). MS m / z (ESI): 423.1 [M+H] + .

[0200] Step 2: Preparation of tert-butyl (4-(2-(2-methoxyethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 4 was replaced with tert-butyl (4-((6-(3-methoxypropionamido)thieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate to give the title compound (480 mg, yield: 46.3%). MS m / z (ESI): 405.1 [M+H] + .

[0201] Step 3: Preparation of 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-(2-methoxyethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 7 was replaced with tert-butyl (4-(2-(2-methoxyethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)carbamate to obtain the title compound (200 mg, yield: 90.7%). MS m / z (ESI): 421.1 [M+H] + .

[0202] Step 4: Preparation of tert-butyl (4-(4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate The synthetic route of Example 1 was applied, and the raw material 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide in step 8 was replaced with 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-(-2-methoxyethyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide to obtain the title compound (84 mg, yield: 42.7%). MS m / z (ESI): 420.2 [M+H] + .

[0203] Step 5: Preparation of 1-(4-aminobutyl)-2-(2-methoxyethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 4) The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl (4-(4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate to obtain the title compound (22 mg, yield: 74.7%). MS m / z (ESI): 320.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 14.49 (br, 1H), 8.55 (br, 2H), 8.17-8.12 (m, 3H),7.55-7.53 (m, 1H), 4.41 (t, J = 8.0 Hz, 2H), 3.85 (t, J = 8.0 Hz, 2H), 3.31 (s,3H), 3.26 (t, J = 8.0 Hz, 2H), 2.84-2.79 (m, 2H), 1.93-1.85 (m, 2H), 1.73-1.66(m, 2H).

[0204] Example 5: Preparation of 1-(4-aminobutyl)-2-(2-ethoxymethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 5)

[0205] [ka] Step 1: Preparation of tert-butyl (4-((6-(2-ethoxyacetamido)thieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate The synthetic route of Example 1 was applied, and the raw material n-pentanoyl chloride in step 3 was replaced with 2-ethoxyacetyl chloride to give the title compound (863 mg, yield: 92.7%). MS m / z (ESI): 423.1 [M+H] + .

[0206] Step 2: Preparation of tert-butyl (4-(2-(ethoxymethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 4 was replaced with tert-butyl (4-((6-(2-ethoxyacetamido)thieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate to give the title compound (517 mg, yield: 50.4%). MS m / z (ESI): 405.2 [M+H] + .

[0207] Step 3: Preparation of 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-(ethoxymethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 7 was replaced with tert-butyl (4-(2-(ethoxymethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate to obtain the title compound (187 mg, yield: 89.3%). MS m / z (ESI): 421.1 [M+H] + .

[0208] Step 4: Preparation of tert-butyl (4-(4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate The synthetic route of Example 1 was applied, and the raw material 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide in step 8 was replaced with 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-(ethoxymethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide to obtain the title compound (76 mg, yield: 38.5%). MS m / z (ESI): 420.2 [M+H] + .

[0209] Step 5: Preparation of 1-(4-aminobutyl)-2-(2-ethoxymethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 5) The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl (4-(4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate to obtain the title compound (33 mg, yield: 81.4%). MS m / z (ESI): 320.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 14.49 (br, 1H), 8.70 (br, 2H), 8.20 (d, J = 4.0 Hz,1H), 8.11 (br, 2H), 7.56 (d, J = 4.0 Hz, 1H), 4.84 (s, 2H), 4.46-4.42 (m, 2H),3.62-3.56 (m, 2H), 2.84-2.79 (m, 2H), 1.98-1.90 (m, 2H), 1.76-1.68 (m, 2H),1.18 (t, J = 8.0 Hz, 3H).

[0210] Example 6: Preparation of 2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 6)

[0211] [ka] Step 1: Preparation of 6-nitrothieno[3,2-b]pyridin-7-amine 7-Chloro-6-nitrothieno[3,2-b]pyridine (2.0 g, 9.23 mmol) was dissolved in methanol (20 mL) and added with ammonia in methanol (7N, 26.4 mL, 184.8 mmol), heated to 50° C. and stirred for 4 h. After cooling to room temperature, the reaction was concentrated to remove the solvent to give the title compound of this step (1.73 g, yield: 96.1%). MS m / z (ESI): 196.1 [M+H] + .

[0212] Step 2: Preparation of thieno[3,2-b]pyridine-6,7-diamine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-nitrothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 2 was replaced with 6-nitrothieno[3,2-b]pyridin-7-amine to give the title compound (1.4 g, yield: 98.4%). MS m / z (ESI): 166.1 [M+H] + .

[0213] Step 3: Preparation of N-(7-aminothieno[3,2-b]pyridin-6-yl)pentanamide The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-aminothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 3 was replaced with thieno[3,2-b]pyridine-6,7-diamine to give the title compound (2.5 g, yield: 97.6%). MS m / z (ESI): 250.1 [M+H] + .

[0214] Step 4: Preparation of 2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine N-(7-aminothieno[3,2-b]pyridin-6-yl)pentanamide (2.5 g, 8.48 mmol) was added to ethanol (30 mL), sodium hydroxide (1.7 g, 42.4 mmol) was added at room temperature, and the mixture was heated to 50° C. and stirred for 6 hours. The reaction was quenched by adding water (150 mL), adjusted to pH=6 with dilute hydrochloric acid, extracted three times with ethyl acetate (150 mL), and the organic phases were combined, washed twice with saturated sodium bicarbonate solution (60 mL), washed twice with saturated saline (60 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated to give the title compound (1.5 g, yield: 76.5%). MS m / z (ESI): 232.0 [M+H] + .

[0215] Step 5: Preparation of 7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 5 was replaced with 2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine to give the title compound (1.4 g, yield: 65.7%). MS m / z (ESI): 310.0 [M+H] + .

[0216] Step 6: Preparation of a mixture of tert-butyl 7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-carboxylate and tert-butyl 7-bromo-2-butyl-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-3-carboxylate 7-Bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine (1 g, 3.20 mmol) and 4-dimethylaminopyridine (40 mg, 0.32 mmol) were dissolved in tetrahydrofuran (10 mL), and triethylamine (0.65 g, 6.40 mmol) and di-tert-butyl dicarbonate (1.1 g, 4.8 mmol) were added successively at room temperature, warmed to 40° C., and stirred for 12 hours. The reaction mixture was poured into water (100 mL), extracted three times with dichloromethane (60 mL), the organic phases were combined, washed three times with saturated saline (30 mL), the organic phase was dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate:petroleum ether=3:1, volume ratio) to obtain the title compound of this step (1.0 g, yield: 76.7%). MS m / z (ESI): 410.0 [M+H] + .

[0217] Step 7: Preparation of a mixture of 7-bromo-1-(tert-butoxycarbonyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide and 7-bromo-3-(tert-butoxycarbonyl)-2-butyl-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide A mixture of tert-butyl 7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-carboxylate and tert-butyl 7-bromo-2-butyl-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-3-carboxylate (1.0 g, 2.43 mmol) was dissolved in dichloromethane (10 mL), cooled in an ice-water bath, and slowly added to the reaction solution at 0° C. along with m-chloroperoxybenzoic acid (0.98 g, 4.86 mmol), slowly warmed to room temperature, and stirred for 4 hours. The reaction mixture was poured into water (100 mL) and extracted three times with dichloromethane (60 mL). The organic phases were combined and washed twice with saturated aqueous sodium bicarbonate solution (30 mL), twice with saturated aqueous sodium sulfite solution (30 mL), and three times with saturated brine (45 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the title compound of this step (1.0 g, yield: 90.9%). MS m / z (ESI): 426.0 [M+H] + .

[0218] Step 8: Preparation of a mixture of tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-carboxylate and tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-3-carboxylate A mixture of 7-bromo-1-(tert-butoxycarbonyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide and 7-bromo-3-(tert-butoxycarbonyl)-2-butyl-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide (1.0 g, 2.34 mmol) and tert-butylamine (0.85 g, 11.7 mmol) was dissolved in dichloromethane (20 mL), cooled in an ice-water bath, and p-toluenesulfonyl chloride (0.89 g, 4.68 mmol) was added slowly in a batch to the reaction mixture at 0° C., slowly heated to room temperature, and stirred for 12 hours. The reaction mixture was added with water (100 mL) and extracted three times with dichloromethane (60 mL). The organic phases were combined and washed three times with saturated saline (45 mL). The organic phase was dehydrated and concentrated over anhydrous sodium sulfate. The resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate:petroleum ether = 5:1, volume ratio) to obtain a mixture of the title compounds of this step (0.6 g, yield: 53.1%). MS m / z (ESI): 481.2 [M+H] + .

[0219] Step 9: Preparation of N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine tert-Butyl 7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-carboxylate and tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-3-carboxylate (0.1 g, 0.21 mmol), 1-methyl-4-(4,4,5, A mixture of 5-tetramethyl-1,3,2-dioxaborocyclopentan-2-yl)-1,2,3,6-tetrahydropyridine (95.0 mg, 0.42 mmol), potassium carbonate (90.0 mg, 0.63 mmol) and 1,1'-bisdiphenylphosphinoferrocene palladium dichloride (15 mg, 0.02 mmol) was added to a mixed solvent of 1,4-dioxane (4 mL) and water (0.5 mL). After blowing nitrogen into the reaction system for 5 minutes, the temperature was raised to 90°C under microwave and stirred for 2 hours. After cooling to room temperature, the reaction system was concentrated to remove the solvent, and the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 10:1, volume ratio) to obtain the title compound (40 mg, yield: 50.0%). MS m / z (ESI): 398.3 [M+H] + .

[0220] Step 10: Preparation of N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (40 mg, 0.1 mmol) was dissolved in methanol (5 mL), 10% palladium on carbon (20 mg, 0.12 mmol) was added, hydrogen substitution was performed three times, the reaction system was warmed to room temperature, stirred for 12 hours, subjected to suction filtration, the filter cake was washed with methanol (5 mL), and the filtrate was concentrated to obtain the title compound (30 mg, yield: 75.0%). MS m / z (ESI): 400.1 [M+H] + .

[0221] Step 11: Preparation of 2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 6) N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (20 mg, 0.05 mmol) was dissolved in methanol (3 mL), concentrated hydrochloric acid (0.5 mL) was added at room temperature, and the mixture was heated to 50° C. and stirred for 4 hours. After cooling to room temperature, the reaction system was concentrated to remove the solvent, and the resulting residue was purified by preparative high performance liquid chromatography to obtain the title compound (18 mg, yield: 94.7%). MS m / z (ESI): 344.1 [M+H] + . 1 H NMR (400 MHz, CD 3 OD): δ 7.31 (s, 1H), 3.70-3.62 (m, 2H), 3.56-3.45 (m, 1H),3.26-3.19 (m, 2H), 3.00-2.96 (m, 2H), 2.94 (s, 3H), 2.42-2.38 (m, 2H),2.12-2.08 (m, 2H), 1.91-1.84 (m, 2H), 1.51-1.42 (m, 2H), 1.00 (t, J = 8.0 Hz,3H).

[0222] Example 7: Preparation of 2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 8)

[0223] [ka]

[0224] The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (25 mg, yield: 78.2%). MS m / z (ESI): 342.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.15 (s, 1H), 8.54 (s, 2H), 7.43 (s, 1H), 6.37 (s,1H), 4.01-3.93 (m, 2H), 3.82 -3.79 (m, 1H), 3.62-3.57 (m, 1H), 3.29-3.27 (m,1H), 2.96-2.81 (m, 7H), 1.82-1.71 (m, 2H), 1.41-1.35 (m, 2H), 0.93 (t, J = 8.0Hz, 3H).

[0225] Example 8: Preparation of 2-butyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 25)

[0226] [ka] Step 1: Preparation of 1-((1-((benzyloxy)carbonyl)piperidin-4-yl)methyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide The synthetic route of Example 1 was applied, and the starting material tert-butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 7 was replaced with benzyl 4-((2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate (250 mg, yield: 85.6%). MS m / z (ESI): 479.2 [M+H] + .

[0227] Step 2: Preparation of benzyl 4-((4-amino-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide in step 8 was replaced with 1-(1-((benzyloxy)carbonyl)piperidin-4-yl)methyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide to obtain the title compound (155 mg, yield: 56.7%). MS m / z (ESI): 478.2 [M+H] + .

[0228] Step 3: Preparation of 2-butyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 25) The synthetic route of Example 3 was applied, and the starting material benzyl 4-((4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate in step 9 was replaced with benzyl 4-((4-amino-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to obtain the title compound (20 mg, yield: 53.8%). MS m / z (ESI): 344.2 [M+H] + . 1 H NMR (400 MHz, CDCl3): δ 7.40 (d, J = 8.0 Hz, 1H), 7.31 (d, J = 8.0 Hz, 1H), 5.19 (s, 2H), 4.09(d, J = 8.0 Hz, 2H), 3.28-3.25 (m, 2H), 2.80-2.76 (m, 2H), 2.63-2.57 (m, 4H),2.11-2.06 (m, 1H), 1.83-1.75 (m, 2H), 1.76- 1.66 (m, 3H), 1.46-1.36 (m, 2H),0.99 (t, J = 8.0 Hz, 3H).

[0229] Example 9: Preparation of 2-butyl-7-methyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 26)

[0230] [ka] Step 1: Preparation of 6-nitro-N-((tetrahydro-2H-pyran-4-yl)methyl)thieno[3,2-b]pyridin-7-amine Applying the synthetic route of Example 1, the raw material N-tert-butoxycarbonyl-1,4-butanediamine in step 1 was replaced with 4-aminomethyltetrahydropyran to give the title compound (0.8 g, yield: 93.6%). MS m / z (ESI): 294.1 [M+H] + .

[0231] Step 2:N 7 Preparation of -((tetrahydro-2H-pyran-4-yl)methyl)thieno[3,2-b]pyridine-6,7-diamine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-nitrothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 2 was replaced with 6-nitro-N-((tetrahydro-2H-pyran-4-yl)methyl)thieno[3,2-b]pyridin-7-amine to give the title compound (0.55 g, yield: 76.6%). MS m / z (ESI): 264.1 [M+H] + .

[0232] Step 3: Preparation of N-(7-(((tetrahydro-2H-pyran-4-yl)methyl)amino)thieno[3,2-b]pyridin-6-yl)pentanamide The synthetic route of Example 1 was applied to convert the raw material tert-butyl (4-((6-aminothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 3 into N 7 -((tetrahydro-2H-pyran-4-yl)methyl)thieno[3,2-b]pyridine-6,7-diamine to give the title compound (0.7 g, yield: 85.3%). MS m / z (ESI): 348.1 [M+H] + .

[0233] Step 4: Preparation of 2-butyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 4 was replaced with N-(7-(((tetrahydro-2H-pyran-4-yl)methyl)amino)thieno[3,2-b]pyridin-6-yl)pentanamide to give the title compound (0.6 g, yield: 82.2%). MS m / z (ESI): 330.1 [M+H] + .

[0234] Step 5: Preparation of 7-bromo-2-butyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 5 was replaced with 2-butyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine to obtain the title compound (0.7 g, yield: 76.1%). MS m / z (ESI): 408.0 [M+H] + .

[0235] Step 6: Preparation of 2-butyl-7-methyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 6 was replaced with 7-bromo-2-butyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine to obtain the title compound (0.15 g, yield: 80.3%). MS m / z (ESI): 344.2 [M+H] + .

[0236] Step 7: Preparation of 2-butyl-7-methyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 7 was replaced with 2-butyl-7-methyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine (0.15 g, yield: 76.4%). MS m / z (ESI): 360.1 [M+H] + .

[0237] Step 8: Preparation of 2-butyl-7-methyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 26) The synthetic route of Example 1 was applied, and the raw material 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide in step 8 was replaced with 2-butyl-7-methyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide to obtain the title compound (12 mg, yield: 12.0%). MS m / z (ESI): 359.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 6.97 (s, 1H), 6.05 (s, 2H), 4.12 (d, J = 8.0 Hz, 2H),3.85-3.82 (m, 2H), 3.23-3.16 (m, 2H), 2.89-2.81 (m, 2H), 2.56 (s, 3H),2.08-2.05 (m, 1H), 1.83-1.75 (m, 2H), 1.52-1.37 (m, 6H), 0.95 (t, J = 8.0 Hz,3H).

[0238] Example 10: Preparation of 1-((1-(2-(2-(2-aminoethoxy)ethoxy)ethyl)piperidin-4-yl)methyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 15)

[0239] [ka] Step 1: Preparation of tert-butyl (2-(2-(2-(4-((4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidin-1-yl)ethoxy)ethyl)carbamate 2-Butyl-7-methyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (60.0 mg, 0.17 mmol), 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl 4-methylbenzenesulfonate (77.0 mg, 0.19 mmol) and N,N-diisopropylethylamine (40.0 mg, 0.31 mmol) were dissolved in N,N-dimethylformamide (2 mL), heated to 70° C., and stirred for 12 hours. The reaction mixture was poured into water (30 mL) and extracted three times with ethyl acetate (30 mL). The organic phases were combined and washed three times with saturated brine (30 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated. The resulting residue was purified by preparative high performance liquid chromatography to obtain the title compound (30.1 mg, yield: 30.0%). MS m / z (ESI): 589.4 [M+H] + .

[0240] Step 2: Preparation of 1-((1-(2-(2-(2-aminoethoxy)ethoxy)ethyl)piperidin-4-yl)methyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 15) The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl (2-(2-(2-(4-((4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidin-1-yl)ethoxy)ethyl)carbamate to obtain the title compound (10.7 mg, yield: 53.1%). MS m / z (ESI): 489.4 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.94 (s, 1H), 8.42-8.24 (m, 4H), 7.30 (s, 1H),4.25-4.23 (m, 2H), 3.86-3.83 (m, 2H), 3.65-3.60 (m, 2H), 3.50-3.48 (m, 2H),3.21-3.18 (m, 2H), 2.95-2.90 (m, 4H), 2.66 (s, 3H), 2.51-2.48 (m, 2H),1.88-1.73 (m, 5H), 1.54-1.42 (m, 2H), 1.33-1.27 (m, 6H), 0.96 (t, J = 8.0 Hz,3H).

[0241] Example 11: Preparation of 1-((1-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)piperidin-4-yl)methyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 16)

[0242] [ka] The synthetic route of Example 10 was applied, and the raw material 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl 4-methylbenzenesulfonate in step 1 was replaced with 2,5,8,11,14,17,20-heptoxan-22-yl 4-methylbenzenesulfonate to give the title compound (25.3 mg, yield: 37.6%). MS m / z (ESI): 680.3 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 6.97-6.95 (m, 1H), 6.05 (s, 2H), 4.10-4.09 (m, 2H),3.50-3.30 (m, 26H), 3.20 (s, 3H), 2.87-2.82 (m, 4H), 2.57-2.55 (m, 3H),2.42-2.39 (m, 2H), 1.85-1.74 (m, 5H), 1.48-1.36 (m, 6H), 0.94 (t, J = 8.0 Hz,3H).

[0243] Example 12: Preparation of 1-(4-((4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidin-1-yl)-3-(2-(2-aminoethoxy)ethoxy)propan-1-one (Compound 17)

[0244] [ka] Step 1: Preparation of tert-butyl (2-(2-(3-(4-((4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidin-1-yl)-3-oxopropoxy)ethoxy)ethyl)carbamate 2-Butyl-7-methyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (40.0 mg, 0.11 mmol), N-tert-butoxycarbonyl-3-[2-(2-aminoethoxy)ethoxy]propionic acid (33.6 mg, 0.12 mmol), triethylamine (48.6 mg, 0.48 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (45.6 mg, 0.12 mmol) were added sequentially to N,N-dimethylformamide (2 mL) and the reaction was stirred at room temperature for 1 h. The reaction mixture was poured into water (30 mL) and extracted three times with ethyl acetate (30 mL). The organic phases were combined and washed three times with saturated brine (30 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated. The residue was purified by preparative high performance liquid chromatography to obtain the title compound (20.7 mg, yield: 30.5%). MS m / z (ESI): 617.4 [M+H] + .

[0245] Step 2: Preparation of 1-(4-((4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidin-1-yl)-3-(2-(2-aminoethoxy)ethoxy)propan-1-one (compound 17) The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl (2-(2-(3-(4-((4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidin-1-yl)-3-oxopropoxy)ethoxy)ethyl)carbamate to obtain the title compound (4.2 mg, yield: 42.8%). MS m / z (ESI): 517.3 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6): δ 8.33 (s, 2H), 6.98-6.97 (m, 1H), 6.09 (s, 2H),4.43-4.40 (m, 2H), 4.12-4.11 (m, 2H), 3.92-3.88 (m, 2H), 3.64-3.60 (m, 2H),3.56-3.54 (m, 2H), 2.92-2.83 (m, 5H), 2.58-2.54 (m, 4H), 2.44- 2.37 (m, 2H),2.08-1.99 (m, 1H), 1.83-1.75 (m, 2H), 1.54-1.17 (m, 8H), 0.95 (t, J = 8.0 Hz,3H).

[0246] Example 13: Preparation of 2-butyl-7-methyl-1-((1-methylpiperidin-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 28)

[0247] [ka] Step 1: Preparation of tert-butyl 4-(((6-nitrothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate Applying the synthetic route of Example 1, the raw material N-tert-butoxycarbonyl-1,4-tert-4-butanediamine in step 1 was replaced with 1-tert-butoxycarbonyl-4-aminomethylpiperidine to give the title compound (29.0 g, yield: 93.9%). MS m / z (ESI): 393.2 [M+H] + .

[0248] Step 2: Preparation of tert-butyl 4-(((6-aminothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-nitrothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 2 was replaced with tert-butyl 4-(((6-nitrothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate to give the title compound (22.0 g, yield: 92.4%). MS m / z (ESI): 363.2 [M+H] + .

[0249] Step 3: Preparation of tert-butyl 4-(((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-aminothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 3 was replaced with tert-butyl 4-(((6-aminothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate to give the title compound (30.0 g, yield: 98.4%). MS m / z (ESI): 447.3 [M+H] + .

[0250] Step 4: Preparation of tert-butyl 4-((2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 4 was replaced with tert-butyl 4-(((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate to give the title compound (19.0 g, yield: 79.5%). MS m / z (ESI): 429.2 [M+H] + .

[0251] Step 5: Preparation of tert-butyl 4-((7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 5 was replaced with tert-butyl 4-((2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to give the title compound (18.0 g, yield: 72.9%). MS m / z (ESI): 507.2 [M+H] + .

[0252] Step 6: Preparation of tert-butyl 4-((2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 6 was replaced with tert-butyl 4-((7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to obtain the title compound (4.0 g, yield: 82.5%). MS m / z (ESI): 443.2 [M+H] + .

[0253] Step 7: Preparation of 1-((1-(tert-butoxycarbonyl)piperidin-4-yl)methyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 7 was replaced with tert-butyl 4-((2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to give the title compound (5.02 g, yield: 89.3%). MS m / z (ESI): 459.3 [M+H] + .

[0254] Step 8: Preparation of tert-butyl 4-((2-butyl-4-(tert-butylamino)-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide in step 8 was replaced with 1-((1-(tert-butoxycarbonyl)piperidin-4-yl)methyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide to obtain the title compound (4.05 g, yield: 75.7%). MS m / z (ESI): 514.3 [M+H] + .

[0255] Step 9: Preparation of N-(tert-butyl)-2-butyl-7-methyl-1-((1-methylpiperidin-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine tert-Butyl 4-((2-butyl-4-(tert-butylamino)-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate (100 mg, 0.19 mmol) was dissolved in tetrahydrofuran (3 mL), lithium aluminum hydride (37.7 mg, 0.99 mmol) was added at room temperature, heated to 70 ° C., and stirred for 4 hours. Sodium sulfate decahydrate (300.0 mg) was added to the reaction solution at room temperature, stirred for 30 minutes, filtered, the filter cake was washed with tetrahydrofuran (10 mL), the filtrate was concentrated, and the residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 10:1, volume ratio) to obtain the title compound (40.3 mg, yield: 49.6%). MS m / z (ESI): 428.3 [M+H] + .

[0256] Step 10: Preparation of 2-butyl-7-methyl-1-((1-methylpiperidin-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 28) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with N-(tert-butyl)-2-butyl-7-methyl-1-((1-methylpiperidin-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (16.7 mg, yield: 54.7%). MS m / z (ESI): 372.2 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6): δ 6.97-6.96 (m, 1H), 6.05 (s, 2H), 4.11-4.09 (m, 2H),3.34 (s, 3H), 2.86-2.82 (m, 2H), 2.75-2.72 (m, 2H), 2.57-2.55 (m, 3H), 1.87-1.71 (m, 5H), 1.45-1.40 (m, 6H), 0.96 (t, J = 8.0 Hz, 3H).

[0257] Example 14: Preparation of 2-butyl-1-methyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 32)

[0258] [ka] Step 1: Preparation of N-methyl-6-nitrothieno[3,2-b]pyridin-7-amine Applying the synthetic route of Example 1, the starting material N-tert-butoxycarbonyl-1,4-butanediamine in step 1 was replaced with methylamine in tetrahydrofuran solution (2.0 M) to give the title compound (760 mg, yield: 77.9%). MS m / z (ESI): 210.0 [M+H] + .

[0259] Step 2:N 7 Preparation of -methylthieno[3,2-b]pyridine-6,7-diamine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-nitrothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 2 was replaced with N-methyl-6-nitrothieno[3,2-b]pyridin-7-amine to give the title compound (620 mg, yield: 97.3%). MS m / z (ESI): 180.1 [M+H] + .

[0260] Step 3: Preparation of N-(7-(methylamino)thieno[3,2-b]pyridin-6-yl)pentanamide The synthetic route of Example 1 was applied to convert the raw material tert-butyl (4-((6-aminothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 3 into N 7 -methylthieno[3,2-b]pyridine-6,7-diamine to give the title compound (1.05 g, yield: 93.7%). MS m / z (ESI): 264.1 [M+H] + .

[0261] Step 4: Preparation of 2-butyl-1-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 4 was replaced with N-(7-(methylamino)thieno[3,2-b]pyridin-6-yl)pentanamide to give the title compound (605 mg, yield: 67.9%). MS m / z (ESI): 246.1 [M+H] + .

[0262] Step 5: Preparation of 7-bromo-2-butyl-1-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 5 was replaced with 2-butyl-1-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine to give the title compound (576 mg, yield: 79.6%). MS m / z (ESI): 324.0 [M+H] + .

[0263] Step 6: Preparation of 7-bromo-2-butyl-1-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 7 was replaced with 7-bromo-2-butyl-1-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine to obtain the title compound (622 mg, yield: 95.3%). MS m / z (ESI): 340.0 [M+H] + .

[0264] Step 7: Preparation of 7-bromo-2-butyl-1-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine The synthetic route of Example 1 was applied, and the raw material 1-(4-((tert-butoxycarbonyl)amino)butyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide in step 8 was replaced with 7-bromo-2-butyl-1-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide to obtain the title compound (534 mg, yield: 56.9%). MS m / z (ESI): 339.0 [M+H] + .

[0265] Step 8: Preparation of 2-butyl-1-methyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine The synthetic route of Example 6 was applied, and the mixture of tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-carboxylate and tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-3-carboxylate was replaced with 7-bromo-2-butyl-1-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine as the raw material for step 9 to obtain the title compound (52 mg, yield: 43.5%). MS m / z (ESI): 356.3 [M+H] + .

[0266] Step 9: Preparation of 2-butyl-1-methyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 32) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 10 was replaced with 2-butyl-1-methyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (12.5 mg, yield: 89.8%). MS m / z (ESI): 358.3 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 8.18 (s, 2H), 7.04 (s, 1H), 3.87 (s, 3H), 3.12-2.84(m, 5H), 2.48-2.35 (m, 5H), 2.07-2.04 (m, 2H), 1.82-1.70 (m, 4H), 1.44-1.37 (m,2H), 0.93 (t, J = 8.0 Hz, 3H).

[0267] Example 15: Preparation of 2-butyl-7-(1-methylpiperidin-4-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 34)

[0268] [ka] Step 1: Preparation of 7-bromo-2-butyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide The synthetic route of Example 6 was applied, and tert-butyl 7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-carboxylate and tert-butyl 7-bromo-2-butyl-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-3-carboxylate were replaced with 7-bromo-2-butyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine as the raw material for step 7 to obtain the title compound (0.65 g, yield: 93.2%). MS m / z (ESI): 424.0 [M+H] + .

[0269] Step 2: Preparation of 7-bromo-N-(tert-butyl)-2-butyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine The synthetic route of Example 6 was applied, and the mixture of 7-bromo-1-(tert-butoxycarbonyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide and 7-bromo-3-(tert-butoxycarbonyl)-2-butyl-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide was replaced with 7-bromo-2-butyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide as the raw material for step 8 to obtain the title compound (0.42 g, yield: 56.5%). MS m / z (ESI): 479.1 [M+H] + .

[0270] Step 3: Preparation of N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine The synthetic route of Example 6 was applied, replacing tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-carboxylate and tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-3-carboxylate with 7-bromo-N-(tert-butyl)-2-butyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to give the title compound (93.8 mg, yield: 67.7%). MS m / z (ESI): 496.2 [M+H] + .

[0271] Step 4: Preparation of N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 10 was replaced with N-(tert-butyl)2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (90.7 mg, yield: 90.0%). MS m / z (ESI): 498.2 [M+H] + .

[0272] Step 5: Preparation of 2-butyl-7-(1-methylpiperidin-4-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 34) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (20.6 mg, yield: 29.7%). MS m / z (ESI): 442.2 [M+H] + . 1 H NMR (400 MHz, CD 3 OD): δ 7.17 (s, 1H), 4.25-4.23 (m, 2H), 3.95-3.92 (m, 2H),3.34-3.29 (m, 4H), 3.18-3.12 (m, 2H), 2.98-2.94 (m, 3H), 2.83 (s, 3H),2.39-2.35 (m, 2H), 2.27-2.01 (m, 3H), 1.94-1.86 (m, 2H), 1.59-1.47 (m, 6H),1.03 (t, J = 8.0 Hz, 3H).

[0273] Example 16: Preparation of 2-butyl-7-(1-methylpiperidin-4-yl)-1-((1-methylpiperidin-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 38)

[0274] [ka] Step 1: Preparation of 7-bromo-1-((1-(tert-butoxycarbonyl)piperidin-4-yl)methyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 7 was replaced with tert-butyl 4-((7-bromo-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to give the title compound (4.1 g, yield: 89.3%). MS m / z (ESI): 523.1 [M+H] + .

[0275] Step 2: Preparation of tert-butyl 4-((7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate The synthetic route of Example 6 was applied, and the raw materials 7-bromo-1-(tert-butoxycarbonyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide and 7-bromo-3-(tert-butoxycarbonyl)-2-butyl-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide in step 8 were replaced with 7-bromo-1-((1-(tert-butoxycarbonyl)piperidin-4-yl)methyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide to obtain the title compound (2.8 g, yield: 64.7%). MS m / z (ESI): 578.2 [M+H] + .

[0276] Step 3: Preparation of tert-butyl 4-((2-butyl-4-(tert-butylamino)-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate The synthetic route of Example 6 was applied, and the raw material tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-carboxylate and tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-3-carboxylate of 9 were replaced with tert-butyl 4-((7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to obtain the title compound (143 mg, yield: 49.2%). MS m / z (ESI): 595.3 [M+H] + .

[0277] Step 4: Preparation of N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1-((1-methylpiperidin-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine The synthetic route of Example 13 was applied, and the raw material tert-butyl 4-((2-butyl-4-(tert-butylamino)-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate in step 9 was replaced with tert-butyl 4-((2-butyl-4-(tert-butylamino)-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to obtain the title compound (110 mg, yield: 64.2%). MS m / z (ESI): 509.3 [M+H] + .

[0278] Step 5: Preparation of N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1-((1-methylpiperidin-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 10 was replaced with N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (60 mg, yield: 62.4%). MS m / z (ESI): 511.4 [M+H] + .

[0279] Step 6: Preparation of 2-butyl-7-(1-methylpiperidin-4-yl)-1-((1-methylpiperidin-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 38) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1-((1-methylpiperidin-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (15 mg, yield: 37.6%). MS m / z (ESI): 455.3 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6) δ 7.01 (s, 1H), 6.05 (s, 2H), 4.13-4.11 (m, 2H),2.86-2.72 (m, 7H), 2.19 (s, 3H), 2.10 (s, 3H), 2.02-1.96 (m, 4H), 1.82-1.65 (m,7H), 1.48-1.37 (m, 6H), 0.94 (t, J = 8.0 Hz, 3H).

[0280] Example 17: Preparation of 2-butyl-1-((1-methylpiperidin-4-yl)methyl)-7-(tetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (Compound 40)

[0281] [ka] Step 1: Preparation of tert-butyl 4-((2-butyl-4-(tert-butylamino)-7-(3,6-dihydro-2H-pyran-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate tert-Butyl 4-((7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate (407 mg, 0.70 mmol), 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (168 mg, 0.80 mmol), potassium carbonate (180 mg, 1.3 mmol) and 1,1'-bisdiphenylphosphinoferrocene palladium dichloride (44 mg, 0.06 mmol) were added sequentially to a mixture of 1,4-dioxane (10 mL) and water (1 mL). The reaction was sparged with nitrogen for 5 min, then heated to 90° C. under microwave and stirred for 2 h. After cooling to room temperature, the reaction system was concentrated to remove the solvent, and the resulting residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 10:1, volume ratio) to obtain the title compound (320 mg, yield: 78.6%). MS m / z (ESI): 582.3 [M+H] + .

[0282] Step 2: Preparation of N-(tert-butyl)-2-butyl-7-(3,6-dihydro-2H-pyran-4-yl)-1-((1-methylpiperidin-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine The synthetic route of Example 13 was applied, and the raw material tert-butyl 4-((2-butyl-4-(tert-butylamino)-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate in step 9 was replaced with tert-butyl 4-((2-butyl-4-(tert-butylamino)-7-(3,6-dihydro-2H-pyran-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to obtain the title compound (280 mg, yield: 69.4%). MS m / z (ESI): 496.3 [M+H] + .

[0283] Step 3: Preparation of N-(tert-butyl)-2-butyl-1-((1-methylpiperidin-4-yl)methyl)-7-(tetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 10 was replaced with N-(tert-butyl)-2-butyl-7-(3,6-dihydro-2H-pyran-4-yl)-1-(1-methylpiperidin-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (98 mg, yield: 78.1%). MS m / z (ESI): 498.3 [M+H] + .

[0284] Step 4: Preparation of 2-butyl-1-((1-methylpiperidin-4-yl)methyl)-7-(tetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 40) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with N-(tert-butyl)-2-butyl-1-((1-methylpiperidin-4-yl)methyl)-7-(tetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (27 mg, yield: 82.3%). MS m / z (ESI): 442.3 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 7.03 (s, 1H), 6.09 (s, 2H), 4.14-4.12 (m, 2H),3.97-3.93 (m, 2H), 3.49-3.43 (m, 2H), 3.18-3.12 (m, 2H), 2.86-2.79 (m, 4H),2.16 (s, 3H), 1.96-1.93 (m, 2H), 1.83-1.70 (m, 6H), 1.46-1.40 (m, 6H), 0.94 (t,J = 8.0 Hz, 3H).

[0285] Example 18: Preparation of 2-butyl-1-[(1-methylhexahydropyridin-4-yl)methyl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 27)

[0286] [ka] 2-Butyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (50.0 mg, 0.15 mmol) was dissolved in methanol (2 mL) and diluted with glacial acetic acid (9.6 mg, 0.16 mmol) and aqueous formaldehyde (H 2 37 wt.% in O, 121.6 mg, 1.5 mmol) was added to the reaction at 0° C. After stirring at 0° C. for 30 min, the reaction was added along with sodium cyanoborohydride (28.3 mg, 0.45 mmol), slowly warmed to room temperature, and stirred for 2 h. The reaction was concentrated, and the resulting residue was purified by preparative high performance liquid chromatography to give the title compound (15.1 mg, yield: 28.0%). MS m / z (ESI): 358.2 [M+H] + . 1 H NMR (400 MHz, CD 3 OD): δ 7.66 (d, J = 5.6 Hz, 1H), 7.29 (d, J = 5.6 Hz, 1H),4.26 (d, J = 7.6 Hz, 2H),3.01-2.84 (m, 4H), 2.25 (s, 3H), 2.02-1.84 (m, 5H),1.70-1.46 (m, 6H), 1.02 (t, J = 7.6 Hz, 3H).

[0287] Example 19: Preparation of 1-(hexahydropyridin-4-ylmethyl)-2-(2-methoxyethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 55)

[0288] [ka] Step 1: Preparation of tert-butyl 4-[({6-[(3-methoxypropionyl)amino]thieno[3,2-b]pyridin-7-yl}amino)methyl]hexahydropyridine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-aminothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 3 was replaced with tert-butyl 4-((6-aminothieno[3,2-b]pyridin-7-yl)amino)methyl)piperidine-1-carboxylate, and the raw material n-pentanoyl chloride was replaced with 3-methoxypropionyl chloride to give the title compound (1.5 g, yield: 95.5%). MS m / z (ESI): 449.2 [M+H] + .

[0289] Step 2: Preparation of tert-butyl 4-{[2-(2-methoxyethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridine-1-carboxylate The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-((6-pentanamidothieno[3,2-b]pyridin-7-yl)amino)butyl)carbamate in step 4 was replaced with tert-butyl 4-[({6-[(3-methoxypropionyl)amino]thieno[3,2-b]pyridin-7-yl}amino)methyl]hexahydropyridine-1-carboxylate to give the title compound (1.1 g, yield: 74.8%). MS m / z (ESI): 431.2 [M+H] + .

[0290] Step 3: Preparation of 2-(2-methoxyethyl)-1-[(1-(tert-butoxycarbonyl)hexahydropyridin-4-yl)methyl]thieno[3,2-b]imidazo[4,5-d]pyridine-5-oxide The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 7 was replaced with tert-butyl 4-{[2-(2-methoxyethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridine-1-carboxylate to obtain the title compound (1.1 g, yield: 96.5%). MS m / z (ESI): 447.2 [M+H] + .

[0291] Step 4: Preparation of tert-butyl 4-{[2-(2-methoxyethyl)-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridine-1-carboxylate The synthetic route of Example 6 was applied, and 7-bromo-1-(tert-butoxycarbonyl)-2-butyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide and 7-bromo-3-(tert-butoxycarbonyl)-2-butyl-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-5-oxide were replaced with 2-(2-methoxyethyl)-1-[(1-(tert-butoxycarbonyl)hexahydropyridin-4-yl)methyl]thieno[3,2-b]imidazo[4,5-d]pyridine-5-oxide as the raw material of step 8 to obtain the title compound (0.52 g, yield: 42.2%). MS m / z (ESI): 502.3 [M+H] + .

[0292] Step 5: Preparation of 1-(hexahydropyridin-4-ylmethyl)-2-(2-methoxyethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (compound 55) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with tert-butyl 4-{[2-(2-methoxyethyl)-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridine-1-carboxylate to obtain the title compound (30 mg, yield: 74.3%). MS m / z (ESI): 346.2 [M+H] + . 1 H NMR (400 MHz, CD3 OD): δ 8.03 (d, J = 5.6 Hz, 1H), 7.49 (d, J = 5.6 Hz, 1H),4.42 (d, J = 7.6 Hz, 2H), 3.97 (t, J = 6.0 Hz, 2H), 3.44-3.42 (m, 2H), 3.38 (s,3H), 3.26 (t, J = 6.0 Hz, 2H), 3.01-2.89 (m, 2H), 2.42-2.31 (m, 1H), 1.92-1.89(m, 2H), 1.78-1.65 (m, 2H).

[0293] Example 20: Preparation of 2-(2-methoxyethyl)-1-[(1-methylhexahydropyridin-4-yl)methyl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 56)

[0294] [ka] The synthetic route of Example 18 was applied, replacing the raw material 2-butyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine with 1-(hexahydropyridin-4-ylmethyl)-2-(2-methoxyethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to give the title compound (18.3 mg, yield: 32.3%). MS m / z (ESI): 360.2 [M+H] + . 1 H NMR (400 MHz, CD 3OD) δ: 7.92 (d, J = 5.6 Hz, 1H), 7.38 (d, J = 5.6 Hz, 1H),4.40 (d, J = 7.6 Hz, 2H), 3.95 (t, J = 6.0 Hz, 2H), 3.50-3.48 (m, 2H), 3.38 (s,3H), 3.24 (t, J = 6.0 Hz, 2H), 3.02-2.95 (m, 2H), 2.84 (s, 3H), 2.37-2.31 (m,1H), 1.96-1.91 (m, 2H), 1.79-1.71 (m, 2H).

[0295] Example 21: Preparation of 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-(1-methylhexahydropyridin-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 36)

[0296] [ka] Step 1: Preparation of tert-butyl-4-{[2-butyl-7-(1-methylhexahydropyridin-4-yl)-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridine-1-carboxylate The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 10 was replaced with tert-butyl 4-((2-butyl-4-(tert-butylamino)-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to obtain the title compound (50 mg, yield: 66.2%). MS m / z (ESI): 597.4 [M+H] + .

[0297] Step 2: Preparation of 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-(1-methylhexahydropyridin-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (compound 36) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with tert-butyl 4-{[2-butyl-7-(1-methylhexahydropyridin-4-yl)-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridine-1-carboxylate to obtain the title compound (20 mg, yield: 64.3%). MS m / z (ESI): 441.3 [M+H] + . 1 H NMR (400 MHz, CD 3 OD) δ: 7.12 (s, 1H), 4.30 (d, J = 7.6 Hz, 2H), 3.54-3.39(m, 4H), 3.26-3.20 (m, 1H), 3.08-2.89 (m, 6H), 2.81 (s, 3H), 2.32-2.28 (m, 3H),2.15-2.02 (m, 2H), 1.96-1.68 (m, 6H), 1.56-1.50 (m, 2H), 1.03 (t, J = 7.6 Hz,3H).

[0298] Example 22: Preparation of 2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}-7-(1-methylhexahydropyridin-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 42)

[0299] [ka] 2-Butyl-1-(hexahydropyridin-4-ylmethyl)-7-(1-methylhexahydropyridin-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (20.0 mg, 0.05 mmol), 2-bromoethyl methyl ether (13.9 mg, 0.1 mmol) and triethylamine (15.2 mg, 0.15 mmol) were added to N,N-dimethylformamide (2 mL) in sequence, heated to 70° C. and stirred for 12 hours. The reaction solution was poured into water (20 mL), extracted three times with ethyl acetate (30 mL), the organic phases were combined, washed twice with saturated saline (10 mL), the organic phase was dehydrated over anhydrous sodium sulfate, concentrated, and the resulting residue was purified by preparative high performance liquid chromatography to obtain the title compound (4.2 mg, yield: 16.8%). MS m / z (ESI): 499.4 [M+H] + . 1 H NMR (400 MHz, CD 3 OD) δ: 7.14 (s, 1H), 4.29 (d, J = 7.6 Hz, 2H), 3.65 (t, J =5.2 Hz, 2H), 3.53-3.43 (m, 4H), 3.37 (s, 3H), 3.27-3.25 (m, 1H), 3.15-3.00 (m, 4H), 2.99-2.91 (m, 2H), 2.83 (s, 3H), 2.75-2.72 (m, 2H), 2.39-2.35 (m, 2H),2.21-1.99 (m, 3H), 1.97-1.86 (m, 2H), 1.81-1.72 (m, 4H), 1.53-1.49 (m, 2H),1.03 (t, J = 7.6 Hz, 3H).

[0300] Example 23: Preparation of 2-butyl-7-(hexahydropyridin-4-yl)-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 41)

[0301] [ka] Step 1: Preparation of 7-bromo-2-butyl-1-(hexahydropyridin-4-ylmethyl)-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl 4-((7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to give the title compound (0.5 g, yield: 95.5%). MS m / z (ESI): 478.1 [M+H] + .

[0302] Step 2: Preparation of 7-bromo-2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridine The synthetic route of Example 22 was applied, and the raw material 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-(1-methylhexahydropyridin-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine was replaced with 7-bromo-2-butyl-1-(hexahydropyridin-4-ylmethyl)-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridine to obtain the title compound (0.42 g, yield: 75.1%). MS m / z (ESI): 536.2 [M+H] + .

[0303] Step 3: Preparation of tert-butyl 4-(2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl)-1,2,3,6-tetrahydropyridine-1-carboxylate The synthetic route of Example 6 was applied, and a mixture of tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-carboxylate and tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-3-carboxylate was used as the raw material for Step 9 to obtain 7-bromo-2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl]phenylmethyl. {ethyl}-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridine, raw material 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborocyclopentan-2-yl)-1,2,3,6-tetrahydropyridine was replaced with tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborocyclopentan-2-yl)-1,2,3,6-tetrahydropyridine-1-carboxylate to obtain the title compound (0.35 g, yield: 70.0%). MS m / z (ESI): 639.5 [M+H] + .

[0304] Step 4: Preparation of tert-butyl 4-(2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl)hexahydropyridine-1-carboxylate The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 10 was replaced with tert-butyl 4-(2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl)-1,2,3,6-tetrahydropyridine-1-carboxylate to obtain the title compound (0.32 g, yield: 91.5%). MS m / z (ESI): 641.5 [M+H] + .

[0305] Step 5: Preparation of 2-butyl-7-(hexahydropyridin-4-yl)-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (compound 41) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with tert-butyl 4-(2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl)hexahydropyridine-1-carboxylate to obtain the title compound (0.22 g, yield: 91.6%). MS m / z (ESI): 485.3 [M+H] + . 1 H NMR (400 MHz, CD 3 OD) δ: 7.35 (s, 1H), 4.36 (d, J = 7.6 Hz, 2H), 3.75-3.73(m, 2H), 3.68-3.61 (m, 3H), 3.60-3.53 (m, 3H), 3.48-3.46 (m, 1H), 3.39 (s, 3H),3.25-3.19 (m, 2H), 3.06-2.95 (m, 4H), 2.37-2.35 (m, 3H), 2.13 -2.09 (m, 2H),2.01-1.81 (m, 6H), 1.55-1.52 (m, 2H), 1.04 (t, J = 7.6 Hz, 3H).

[0306] Example 24: Preparation of 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 51)

[0307] [ka] Step 1: Preparation of benzyl 4-{2-butyl-4-(tert-butylamino)-1-[(1-((tert-butyloxy)carbonyl)hexahydropyridin-4-yl)methyl]thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}-1,2,3,6-tetrahydropyridine-1-carboxylate The synthetic route of Example 6 was applied, and a mixture of tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-carboxylate and tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-3-carboxylate was used as the raw material for Step 9. 4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate and 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborocyclopentan-2-yl)-1,2,3,6-tetrahydro-pyridine was replaced with benzyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborocyclopentan-2-yl)-1,2,3,6-tetrahydropyridine-1-carboxylate to give the title compound (0.81 g, yield: 69.5%). MS m / z (ESI): 715.4 [M+H] + .

[0308] Step 2: Preparation of tert-butyl 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridine-1-carboxylate Benzyl 4-{2-butyl-4-(tert-butylamino)-1-[(1-((tert-butyloxy)carbonyl)hexahydropyridin-4-yl)methyl]thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}-1,2,3,6-tetrahydropyridine-1-carboxylate (0.8 g, 1.12 mmol), ammonium formate (2.12 g, 33.6 mmol) and 10% palladium on carbon (100 mg, 0.094 mmol) were added successively to absolute ethanol (20 mL), heated to 80° C. and stirred for 12 hours. After cooling to room temperature, the reaction system was suction filtered, the filter cake was washed with ethanol (15 mL), the filtrate was concentrated, the residue was added together with water (100 mL), extracted three times with ethyl acetate (60 mL), the organic phase was combined, washed three times with saturated saline solution (30 mL), the organic phase was dehydrated over anhydrous sodium sulfate, concentrated, and the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 10:1, volume ratio) to obtain the title compound (0.55 g, yield: 84.6%). MS m / z (ESI): 583.4 [M+H] + .

[0309] Step 3: Preparation of tert-butyl 4-({2-butyl-4-(tert-butylamino)-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridine-1-carboxylate Triethylene glycol monomethyl ether (0.5 g, 3.05 mmol) and Dess-Martin oxidant (1.94 g, 4.57 mmol) were added to anhydrous dichloromethane (10 mL) in order, and the reaction system was stirred at room temperature for 12 hours. The reaction solution was slowly added to a mixture of saturated ammonium thiosulfate solution and saturated sodium bicarbonate solution (20 mL, volume ratio 1:1), the mixture was stirred at room temperature for 0.5 hours, extracted three times with dichloromethane (15 mL), the organic phase was combined, washed three times with saturated saline (15 mL), the organic phase was dehydrated with anhydrous sodium sulfate and concentrated to obtain crude 2-(2,5,8-trioxaoctyl)acetaldehyde (0.45 g, yield: 90%).

[0310] In a separate reaction flask, tert-butyl 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridine-1-carboxylate (0.3 g, 0.51 mmol), sodium cyanoborohydride (161.8 mmol, 2.6 mmol) and glacial acetic acid (6 mg, 0.1 mmol) were added sequentially to anhydrous methanol (4 mL). The above 2-(2,5,8-trioxafoctyl)acetaldehyde (130 mg, 0.78 mmol) in anhydrous methanol solution (1 mL) was slowly added dropwise at room temperature and the reaction was stirred at room temperature for 2 hours. The reaction mixture was poured into water (50 mL) and extracted three times with dichloromethane (30 mL). The organic phases were combined and washed three times with saturated saline (15 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 10:1, volume ratio) to obtain the title compound (0.22 g, yield: 58.6%). MS m / z (ESI): 729.5 [M+H] + .

[0311] Step 4: Preparation of 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (compound 51) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with tert-butyl 4-({2-butyl-4-(tert-butylamino)-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridine-1-carboxylate to obtain the title compound (132 mg, yield: 75.8%). MS m / z (ESI): 573.4 [M+H] + . 1 H NMR (400 MHz, CD 3 OD) δ: 7.13 (s, 1H), 4.30 (d, J = 7.6 Hz, 2H), 3.82 (t, J =5.2 Hz, 2H), 3.71-3.62 (m, 6H), 3.60-3.52 (m, 4H), 3.43-3.38 (m, 2H), 3.36 (s,3H), 3.20-3.17 (m, 3H), 3.01-2.88 (m, 6H), 2.31-2.28 (m, 3H), 2.08-2.05 (m,2H), 1.97-1.81 (m, 4H), 1.68-1.66 (m, 2H), 1.56-1.50 (m, 2H), 1.03 (t, J = 7.6Hz, 3H).

[0312] Example 25: Preparation of 2-butyl-1-[(1-methylhexahydropyridin-4-yl)methyl]-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 52)

[0313] [ka] Applying the synthetic route of example 18, 2-butyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine was replaced with 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to give the title compound (7.2 mg, yield: 22.5%). MS m / z (ESI): 587.4 [M+H] + . 1 H NMR (400 MHz, CD 3 OD) δ: 7.08 (s, 1H), 4.24 (d, J = 7.6 Hz, 2H), 3.82-3.53(m, 10H), 3.38-3.35 (m, 4H), 3.15-2.85 (m, 8H), 2.63-2.59 (m, 2H), 2.39 (s,3H), 2.27-1.84 (m, 9H), 1.74-1.45 (m, 6H), 1.02 (t, J = 7.6 Hz, 3H).

[0314] Example 26: Preparation of 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-[1-(2,5,8,11,14-pentaoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 57)

[0315] [ka] Step 1: Preparation of tert-butyl 4-({2-butyl-4-(tert-butylamino)-7-[1-(2,5,8,11,14-pentaoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridine-1-carboxylate Applying the synthetic route of Example 24, the raw material triethylene glycol monomethyl ether in step 3 was replaced with 2,5,8,11,14-pentoxa-16-hexadecanol to give the title compound (0.35 g, yield: 54.5%). MS m / z (ESI): 817.5 [M+H] + .

[0316] Step 2: Preparation of 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-[1-(2,5,8,11,14-pentaoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (compound 57) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with tert-butyl 4-({2-butyl-4-(tert-butylamino)-7-[1-(2,5,8,11,14-pentaoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridine-1-carboxylate to obtain the title compound (0.16 g, yield: 82.4%). MS m / z (ESI): 661.5 [M+H] + . 1 H NMR (400 MHz, CD 3OD) δ: 7.11 (s, 1H), 4.30 (d, J = 7.6 Hz, 2H), 3.78 (t, J =5.2 Hz, 2H), 3.70-3.54 (m, 16H), 3.51-3.36 (m, 7H), 3.16-3.05 (m, 3H),2.99-2.86 (m, 4H), 2.78-2.70 (m, 2H), 2.35-2.20 (m, 3H), 2.09-1.79 (m, 6H),1.74-1.62 (m, 2H), 1.55-1.50 (m, 2H), 1.03 (t, J = 7.6 Hz, 3H).

[0317] Example 27: Preparation of 2-butyl-1-[(1-methylhexahydropyridin-4-yl)methyl]-7-[1-(2,5,8,11,14-pentoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 58)

[0318] [ka] The synthetic route of Example 18 was applied, replacing 2-butyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine with 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-[1-(2,5,8,11,14-trioxadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine, to give the title compound (9.2 mg, yield: 31.3%). MS m / z (ESI): 675.5 [M+H] + . 1 H NMR (400 MHz, CD 3OD) δ: 7.07 (s, 1H), 4.23 (d, J = 7.6 Hz, 2H), 3.75-3.47(m, 22H), 3.38-3.32 (m, 2H), 3.25-3.22 (m, 2H), 3.05-2.90 (m, 3H), 2.81-2.77(m, 2H), 2.48-2.44 (m, 2H), 2.31 (s, 3H), 2.20-1.84 (m, 8H), 1.66 -1.48 (m,6H), 1.02 (t, J = 7.6 Hz, 3H).

[0319] Example 28: Preparation of 2-butyl-7-(hexahydropyridin-4-yl)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 33)

[0320] [ka] Step 1: Preparation of tert-butyl-4-{2-butyl-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}-1,2,3,6-tetrahydropyridine-1-carboxylate 7-Bromo-N-(tert-butyl)-2-butyl-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (8.2 g, 17.10 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborocyclopentan-2-yl)-1,2,3,6-tetrahydropyridine-1-carboxylate (7.93 g, 25.7 mmol), potassium carbonate (4.73 g, 34.2 mmol) and 1,1'-bisdiphenylphosphinoferrocene palladium dichloride (620.4 mg, 0.86 mmol) were added in sequence to a mixed solvent of 1,4-dioxane (120 mL) and water (10 mL). The reaction system was bubbled with nitrogen for 5 minutes, then heated to 100° C. and stirred for 2 hours. After cooling to room temperature, the reaction system was concentrated to remove the solvent, and the resulting residue was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 1:3, volume ratio) to obtain the title compound (4.01 g, yield: 40.3%). MS m / z (ESI): 582.3 [M+H] + .

[0321] Step 2: Preparation of tert-butyl-4-{2-butyl-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridine-1-carboxylate Tert-butyl 4-{2-butyl-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}-1,2,3,6-tetrahydropyridine-1-carboxylate (0.5 g, 0.86 mmol), ammonium formate (1.08 g, 17.2 mmol) and 10% palladium on carbon (20 mg, 0.019 mmol) were added sequentially to anhydrous ethanol (10 mL), slowly heated to 80° C. and stirred for 12 hours. After cooling to room temperature, the reaction system was filtered by suction, the filter cake was washed with ethanol (15 mL), the filtrate was concentrated, and the residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 10:1, volume ratio) to obtain the title compound (0.40 g, yield: 79.7%). MS m / z (ESI): 584.3 [M+H] + .

[0322] Step 3: Preparation of 2-butyl-7-(hexahydropyridin-4-yl)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (compound 33) tert-Butyl 4-{2-butyl-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridine-1-carboxylate (0.3 g, 0.51 mmol) was dissolved in methanol (3 mL), concentrated hydrochloric acid (0.5 mL) was added at room temperature, and the mixture was heated to 50° C. and stirred for 4 hours. After cooling to room temperature, the reaction system was concentrated to remove the solvent, and the resulting residue was purified by preparative high performance liquid chromatography to obtain the title compound (0.19 g, yield: 87.1%). MS m / z (ESI): 428.3 [M+H] + . 1 H NMR (400 MHz, CD 3OD) δ: 7.11 (s, 1H), 4.23 (d, J = 7.6 Hz, 2H), 3.95-3.92(m, 2H), 3.47-3.44 (m, 2H), 3.15-3.08 (m, 2H), 2.98-2.94 (m, 2H), 2.31-2.15 (m,3H), 2.02-1.86 (m, 4H), 1.57-1.49 (m, 5H), 1.37-1.26 (m, 4H), 1.03 (t, J = 7.6Hz, 3H).

[0323] Example 29: Preparation of 1-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}ethyl-1-one (Compound 59)

[0324] [ka] 2-Butyl-7-(hexahydropyridin-4-yl)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (50 mg, 0.12 mmol) and triethylamine (18 mg, 0.18 mmol) were dissolved in anhydrous dichloromethane (2 mL), and added with acetic anhydride (13 mg, 0.13 mmol) at 0° C. and stirred for 2 hours at 0° C. The reaction solution was concentrated to remove the solvent, and the resulting residue was purified by preparative high performance liquid chromatography to obtain the title compound (37 mg, yield: 65.7%). MS m / z (ESI): 470.2 [M+H] + . 1 H NMR (400 MHz, CD 3OD) δ: 7.11 (s, 1H), 4.67-4.63 (m, 1H), 4.23-4.22 (m, 2H),4.07-4.03 (m, 1H), 3.95-3.92 (m, 2H), 3.36-3.22 (m, 4H), 2.95 (t, J = 8.0 Hz,2H), 2.82-2.76 (m, 1H), 2.22-2.11 (m, 6H), 1.92-1.86 (m, 2H), 1.82-1.61 (m,2H), 1.57-1.47 (m, 6H), 1.02 (t, J = 8.0 Hz, 3H).

[0325] Example 30: Preparation of 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-(3,4,5,6-tetrahydro-2H-pyran-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 39)

[0326] [ka] Step 1: Preparation of tert-butyl 4-({2-butyl-4-(tert-butylamino)-7-(3,4,5,6-tetrahydro-2H-pyran-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridine-1-carboxylate The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 10 was replaced with tert-butyl 4-((2-butyl-4-(tert-butylamino)-7-(3,6-dihydro-2H-pyran-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)methyl)piperidine-1-carboxylate to obtain the title compound (150 mg, yield: 93.2%). MS m / z (ESI): 584.4 [M+H] + .

[0327] Step 2: Preparation of 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-(3,4,5,6-tetrahydro-2H-pyran-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (compound 39) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with tert-butyl 4-({2-butyl-4-(tert-butylamino)-7-(3,4,5,6-tetrahydro-2H-pyran-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridine-1-carboxylate to obtain the title compound (25 mg, yield: 38.1%). MS m / z (ESI): 428.3 [M+H] + .

[0328] Example 31: Preparation of 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-[1-(2-methoxyethyl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (Compound 47)

[0329] [ka] Step 1: Preparation of tert-butyl 4-({2-butyl-7-[1-(2-methoxyethyl)hexahydropyridin-4-yl]-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridine-1-carboxylate Applying the synthetic route of Example 24, the raw material triethylene glycol monomethyl ether in step 3 was replaced with 2-methoxyethanol to give the title compound (260 mg, yield: 76.3%). MS m / z (ESI): 641.4 [M+H] + .

[0330] Step 2: Preparation of 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-[1-(2-methoxyethyl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (compound 47) The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with tert-butyl 4-({2-butyl-7-[1-(2-methoxyethyl)hexahydropyridin-4-yl]-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridine-1-carboxylate to obtain the title compound (34 mg, yield: 42.3%). MS m / z (ESI): 485.3 [M+H] + .

[0331] II. Preparation of conjugate intermediates Example 2-1: Preparation of N-(4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)-4-((2,5-dioxy-2,5-dihydro-1H-pyrrol-1-yl)methyl)cyclohexylcarboxamide (compound D-L'-1)

[0332] [ka] 1-(4-aminobutyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (compound 1) (1.50 g, 4.53 mmol) was dissolved in a mixed solvent of dichloromethane (10 mL) and N,N-dimethylformamide (10 mL), cooled in an ice-water bath, and 4-(N-maleimidomethyl)cyclohexyl-1-carboxylic acid succinimidyl ester (1.82 g, 5.43 mmol) and N,N-diisopropylethylamine (1.17 g, 9.06 mmol) were added to the reaction solution at 0° C., slowly warmed to room temperature, and stirred for 12 hours. The reaction mixture was poured into water (150 mL) and extracted three times with dichloromethane (100 mL). The organic phases were combined and washed three times with saturated saline (100 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 10:1, volume ratio) to obtain the title compound (1.55 g, yield: 62.1%). MS m / z (ESI): 551.3 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 8.15 (s, 1H), 7.02 (s, 2H), 6.96-6.94 (m, 1H), 6.08(s, 2H), 4.19 (t, J = 8.0 Hz, 2H), 3.23-3.21 (m, 3H), 3.08-3.03 (m, 2H),2.86-2.82 (m, 2H), 2.55-2.52 (m, 3H), 1.95-1.91 (m, 1H), 1.80 -1.70 (m, 4H),1.63-1.40 (m, 8H), 1.23-1.16 (m, 2H), 0.95 (t, J = 8.0 Hz, 3H), 0.90-0.79 (m,2H).

[0333] Example 2-2: Preparation of N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-({4-[({6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]-1-oxohex-5-ynyl}amino)methyl]-1,2,3-triazol-1-yl}methyl)cyclohexylcarboxamide (compound D-L'-2)

[0334] [ka] Step 1: Preparation of N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-(hydroxymethyl)cyclohexylcarboxamide 1-(4-aminobutyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine (0.5 g, 1.51 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.69 g, 1.81 mmol), 4-(hydroxymethyl)cyclohexyl-1-carboxylic acid (0.24 g, 1.51 mmol) and N,N-diisopropylethylamine (0.39 g, 3.02 mmol) were added to anhydrous N,N-dimethylformamide (10 mL) in that order and stirred at room temperature for 12 hours. The reaction mixture was poured into water (30 mL) and extracted three times with dichloromethane (30 mL). The organic phases were combined and washed three times with saturated saline (15 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 10:1, volume ratio) to obtain the title compound (0.61 g, yield: 85.6%). MS m / z (ESI): 472.3 [M+H] + .

[0335] Step 2: Preparation of 4-({[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]amino}carbonyl)cyclohexyl]methyl 4-methylbenzenesulfonate N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-(hydroxymethyl)cyclohexylcarboxamide (0.5 g, 1.06 mmol), 4-dimethylaminopyridine (13 mg, 0.11 mmol), N,N-diisopropylethylamine (0.27 g, 2.12 mmol) and p-toluenesulfonyl chloride (0.3 g, 1.59 mmol) were added successively to anhydrous N,N-dimethylformamide (10 mL) and stirred at room temperature for 12 hours. The reaction mixture was poured into water (30 mL) and extracted three times with dichloromethane (30 mL). The organic phases were combined and washed three times with saturated saline (15 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 10:1, volume ratio) to obtain the title compound (0.31 g, yield: 46.7%). MS m / z (ESI): 626.3 [M+H] + .

[0336] Step 3: Preparation of N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-(azidomethyl)cyclohexylcarboxamide [4-({[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]amino}carbonyl)cyclohexyl]methyl 4-methylbenzenesulfonate (0.15 g, 0.24 mmol) and sodium azide (31.2 mg, 0.48 mmol) were added successively to anhydrous N,N-dimethylformamide (3 mL), heated to 70° C., and stirred for 12 hours. The reaction solution was poured into water (30 mL), extracted three times with dichloromethane (30 mL), the organic phase was combined, washed three times with saturated saline (15 mL), and the organic phase was dehydrated over anhydrous sodium sulfate and concentrated to obtain the title compound (110 mg, yield: 92.3%). MS m / z (ESI): 497.3 [M+H] + .

[0337] Step 4: Preparation of N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-({4-[({6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]-1-oxohex-5-ynyl}amino)methyl]-1,2,3-triazol-1-yl}methyl)cyclohexylcarboxamide N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-(azidomethyl)cyclohexylcarboxamide (100mg, 0.20mmol), 6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]-N-(prop-1-ynyl-3-yl)hex-5-ynylamide (68mg, 0.22mmol), and sodium ascorbate (59mg, 0.30mmol) and anhydrous copper sulfate (64mg, 0.40mmol) were added in sequence to a mixed solvent of dimethyl sulfoxide (2mL) and water (0.5mL), and the mixture was stirred at room temperature for 1 hour. The reaction mixture was poured into water (30 mL) and extracted three times with dichloromethane (30 mL). The organic phases were combined and washed three times with saturated brine (15 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated. The resulting residue was purified by preparative high performance liquid chromatography to obtain the title compound (30 mg, yield: 18.7%). MS m / z (ESI): 802.4 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.10 (s, 2H), 8.39 (s, 1H), 7.88 (s, 1H), 7.70 (s,1H), 6.94 (s, 1H), 6.03 (s, 2H), 4.29-4.17 (m, 6H), 3.41-3.35 (m, 6H), 3.05-2.84(m, 4H), 2.30 (s, 2H), 1.91-1.23 (m, 20H), 1.22-0.93 (m, 5H).

[0338] Example 2-3: Preparation of N-{26-[4-(4-amino-2-butyl-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hexa-5-ynamide (compound D-L'-3)

[0339] [ka] Step 1: Preparation of tert-butyl-4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}-1,2,3,6-tetrahydropyridine-1-carboxylate The synthetic route of Example 6 was applied, and the raw material 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborocyclopentan-2-yl)-1,2,3,6-tetrahydropyridine in step 9 was replaced with tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborocyclopentan-2-yl)-1,2,3,6-tetrahydropyridine-1-carboxylate to give the title compound (0.32 g, yield: 56.8%). MS m / z (ESI): 484.3 [M+H] + .

[0340] Step 2: Preparation of tert-butyl-4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridine-1-carboxylate The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 10 was replaced with tert-butyl 4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}-1,2,3,6-tetrahydropyridine-1-carboxylate to obtain the title compound (0.25 g, yield: 83.3%). MS m / z (ESI): 486.3 [M+H] + .

[0341] Step 3: Preparation of 2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl 4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridine-1-carboxylate to obtain the title compound (0.16 g, yield: 80.8%). MS m / z (ESI): 386.3 [M+H] + .

[0342] Step 4: Preparation of tert-butyl {[26-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]amino}carboxylate The synthetic route of Example 24 was applied, and the raw material triethylene glycol monomethyl ether in step 3 was replaced with tert-butyl [(26-hydroxy-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl) amino] carboxylate, and the raw material tert-butyl 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino) thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl) methyl] hexahydropyridine-1-carboxylate was replaced with 2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridine to obtain the title compound (80 mg, yield: 45.2%). MS m / z (ESI): 881.6 [M+H] + .

[0343] Step 5: Preparation of 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosan-1-yl]oxa}ethyl-1-amine The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with tert-butyl {[26-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]amino}carboxylate to obtain the title compound (62 mg, yield: 75.3%). MS m / z (ESI): 725.4 [M+H] + .

[0344] Step 6: Preparation of N-{26-[4-(4-amino-2-butyl-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosan-1-yl]oxy}ethyl-1-amine (50 mg, 64.0 μmol), N,N-diisopropylethylamine (16.5 mg, 128.0 μmol) and 1-({1-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]-6-oxohex-1-ynyl-6-yl}oxy)tetrahydropyrrole-2,5-dione (28.1 mg, 77 μmol) were added sequentially to N,N-dimethylformamide (2 mL) and stirred at room temperature for 1 hour. The reaction mixture was purified by preparative high performance liquid chromatography to obtain the title compound (20 mg, yield: 32.0%). MS m / z (ESI): 975.5 [M+H] + . 1 H NMR (400 MHz, D2 O): δ 8.75 (s, 2H), 8.50 (s, 1H), 7.07 (s, 1H), 3.96-3.93(m, 2H), 3.86-3.46 (m, 35H), 3.43 (s, 3H), 3.42-3.25 (m, 4H), 2.91 (t, J = 7.6Hz, 2H), 2.57-2.41 (m, 6H), 2.11 (s, 2H), 1.96-1.75 (m, 4H), 1.44-1.37 (m, 2H),0.97 (t, J = 7.6 Hz, 3H).

[0345] Example 2-4: Preparation of N-(26-{4-[(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-4)

[0346] [ka] Step 1: Preparation of tert-butyl [(26-{4-[(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)amino]carboxylate The synthetic route of Example 24 was applied, and the raw material triethylene glycol monomethyl ether in step 3 was replaced with tert-butyl [(26-hydroxy-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl) amino] carboxylate, and the raw material tert-butyl 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino) thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl) methyl] hexahydropyridine-1-carboxylate was replaced with 2-butyl-7-methyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (70 mg, yield: 52.3%). MS m / z (ESI): 853.5 [M+H] + .

[0347] Step 2: Preparation of 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-4-amine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl [(26-{4-[(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)amino]carboxylate to obtain the title compound (55 mg, yield: 85.6%). MS m / z (ESI): 753.5 [M+H] + .

[0348] Step 3: Preparation of N-(26-{4-[(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}-3,6,9,12,15,18,21,24-octoxahexacosan-1-yl)-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Examples 2-3 was applied, and the reaction material 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosane-1-yl]oxy}ethyl-1-amine in step 6 was replaced with 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to obtain the title compound (19 mg, yield: 47.2%). MS m / z (ESI): 1003.6 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.12 (s, 2H), 8.00-7.97 (m, 1H), 7.05 (s, 1H),6.69-6.52 (br, 2H), 4.17 (d, J = 7.6 Hz, 2H), 3.61-3.47 (m, 36H), 2.92-2.81 (m,4H), 2.62-2.54 (m, 8H), 2.27 (t, J = 7.6 Hz, 2H), 2.17-2.04 (m, 1H), 1.87-1.64(m, 8H), 1.45-1.42 (m, 2H), 1.27-1.22 (m, 2H), 0.96 (t, J = 7.6 Hz, 3H).

[0349] Example 2-5: Preparation of N-(26-{4-[(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide (compound D-L'-5)

[0350] [ka] 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-4-amine (20 mg, 26.6 μmol), N,N-diisopropylethylamine (7 mg, 53.2 μmol) and 1-{6-[(2,5-dioxotetrahydro-1H-pyrrol-1-yl)oxy]-6-oxohexyl}pyrrole-2,5-dione (10 mg, 31.9 μmol) were added to N,N-dimethylformamide (1 mL) in sequence and stirred at room temperature for 1 hour. The reaction solution was purified by preparative high performance liquid chromatography to obtain the title compound (11 mg, yield: 43.7%). MS m / z (ESI): 946.5 [M+H] + . 1 H NMR (400 MHz, CD 3 OD): δ 6.99 (s, 1H), 4.24-4.21 (m, 2H), 3.69 (s, 2H),3.65-3.53 (m, 31H), 3.53-3.44 (m, 4H), 3.34-3.32 (m, 2H), 2.98-2.84 (m, 4H),2.61 (s, 3H), 2.39-2.32 (m, 2H), 2.17-2.08 (m, 4H), 1.91-1.85 (m, 2H),1.76-1.48 (m, 10H), 1.33-1.23 (m, 2H), 1.03 (t, J = 7.6 Hz, 3H).

[0351] Example 2-6: Preparation of N-(26-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-6)

[0352] [ka] Step 1: Preparation of tert-butyl-4-{2-butyl-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}-1,2,3,6-tetrahydropyridine-1-carboxylate The synthetic route of Example 6 was applied, and a mixture of tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-carboxylate and tert-butyl 7-bromo-2-butyl-4-(tert-butylamino)-3H-imidazo[4,5-d]thieno[3,2-b]pyridine-3-carboxylate was used as the raw material for step 9. (4,4,5,5-tetramethyl-1,3,2-dioxaborocyclopentan-2-yl)-1,2,3,6-tetrahydropyridine was replaced with tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborocyclopentan-2-yl)-1,2,3,6-tetrahydropyridine-1-carboxylate to give the title compound (0.25 g, yield: 72.8%). MS m / z (ESI): 582.3 [M+H]+ .

[0353] Step 2: Preparation of tert-butyl-4-{2-butyl-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridine-1-carboxylate The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 10 was replaced with tert-butyl 4-{2-butyl-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}-1,2,3,6-tetrahydropyridine-1-carboxylate to obtain the title compound (0.22 g, yield: 88.3%). MS m / z (ESI): 584.4 [M+H] + .

[0354] Step 3: Preparation of 2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl 4-{2-butyl-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridine-1-carboxylate to obtain the title compound (0.13 g, yield: 81.2%). MS m / z (ESI): 484.3 [M+H] + .

[0355] Step 4: Preparation of tert-butyl {[26-(4-{2-butyl-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxohexacosan-1-yl]amino}carboxylate Applying the synthetic route of Example 24, the starting material triethylene glycol monomethyl ether in step 3 was replaced with tert-butyl [(26-hydroxy-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)amino]carboxylate to give the starting material tert-butyl 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino)thieno[3, 2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridine-1-carboxylate was replaced with 2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridine to give the title compound (52 mg, yield: 46.3%). MS m / z (ESI): 979.6 [M+H] + .

[0356] Step 5: Preparation of 7-[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine The synthetic route of Example 6 was applied, and the raw material N-(tert-butyl)-2-butyl-7-(1-methylpiperidin-4-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 11 was replaced with tert-butyl {[26-(4-{2-butyl-4-(tert-butylamino)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octoxahexacosan-1-yl]amino}carboxylate to obtain the title compound (40 mg, yield: 90.9%). MS m / z (ESI): 823.5 [M+H] + .

[0357] Step 6: Preparation of N-(26-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Examples 2-3 was applied to convert the reactant 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosan-1-yl]oxy}ethyl-1-amine in Step 6 into 7-[1- (26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to give the title compound (19 mg, yield: 37.1%). MS m / z (ESI): 1073.5 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.12 (s, 2H), 8.17 (s, 1H), 7.96 (s, 1H), 7.02 (s,1H), 6.08 (s, 2H), 4.13 (d, J = 7.6 Hz, 2H), 3.85-3.82 (m, 2H), 3.57-3.30 (m,31H), 3.26-3.15 (m, 6H), 3.03-3.00 (m, 2H), 2.92-2.82 (m, 3H), 2.60-2.55 (m,5H), 2.31-2.17 (m, 8H), 1.84-1.65 (m, 6H), 1.48-1.38 (m, 6H), 0.94 (t, J = 7.2Hz, 3H).

[0358] Example 2-7: Preparation of N-{26-[4-(4-amino-2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-7)

[0359] [ka] Step 1: Preparation of tert-butyl ({26-[4-(4-amino-2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}amino)carboxylate The synthetic route of Example 24 was applied, and the raw material triethylene glycol monomethyl ether in step 3 was replaced with tert-butyl [(26-hydroxy-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl) amino] carboxylate, and the raw material tert-butyl 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino) thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl) methyl] hexahydropyridine-1-carboxylate was replaced with 2-butyl-7-(hexahydropyridin-4-yl)-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to obtain the title compound (45 mg, yield: 43.3%). MS m / z (ESI): 980.6 [M+H] + .

[0360] Step 2: Preparation of 7-[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]-2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl ({26-[4-(4-amino-2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}amino)carboxylate to obtain the title compound (33 mg, yield: 82.5%). MS m / z (ESI): 880.5 [M+H] + .

[0361] Step 3: Preparation of N-{26-[4-(4-amino-2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Examples 2-3 was applied to convert the reactant material 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosan-1-yl]oxy}ethyl-1-amine in Step 6 into 7-[1-(2 6-amino-3,6,9,12,15,18,21,24-octaoxahexadecan-1-yl)hexahydropyridin-4-yl]-2-butyl-1-{[1-(2-methoxyethyl)hexahydropyridin-4-yl]methyl}thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine was replaced to give the title compound (10 mg, yield: 23.8%). MS m / z (ESI): 1130.6 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.12 (s, 2H), 7.95 (s, 1H), 7.01 (s, 1H), 6.05 (s,2H), 4.14-4.08 (m, 2H), 3.55 -3.45 (m, 22H), 3.43-3.37 (m, 14H), 3.23-3.15 (m,7H), 2.99-2.97 (m, 2H), 2.89-2.82 (m, 4H), 2.45-2.32 (m, 3H), 2.26 (t, J = 7.6Hz, 2H), 2.16-1.94 (m, 8H), 1.85-1.65 (m, 8H), 1.49-1.36 (m, 6H), 0.94 (t, J =7.6 Hz, 3H).

[0362] Example 2-8: Preparation of N-[26-(4-{[4-amino-2-butyl-7-(3,4,5,6-tetrahydro-2H-pyran-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxohexacosan-1-yl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-8)

[0363] [ka] Step 1: Preparation of tert-butyl {[26-(4-{[4-amino-2-butyl-7-(3,4,5,6-tetrahydro-2H-pyran-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]amino}carboxylate The synthetic route of Example 24 was applied, and the raw material triethylene glycol monomethyl ether in step 3 was replaced with tert-butyl [(26-hydroxy-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl) amino] carboxylate, and the raw material tert-butyl 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl) methyl]hexahydropyridine-1-carboxylate was replaced with 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-(3,4,5,6-tetrahydro-2H-pyran-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to obtain the title compound (115 mg, yield: 54.3%). MS m / z (ESI): 923.6 [M+H] + .

[0364] Step 2: Preparation of 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-(3,4,5,6-tetrahydro-2H-pyran-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl {[26-(4-{[4-amino-2-butyl-7-(3,4,5,6-tetrahydro-2H-pyran-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octoxahexacosan-1-yl]amino}carboxylate to obtain the title compound (90 mg, yield: 88.2%). MS m / z (ESI): 823.5 [M+H] + .

[0365] Step 3: Preparation of N-[26-(4-{[4-amino-2-butyl-7-(3,4,5,6-tetrahydro-2H-pyran-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Examples 2 and 3 was applied to convert the raw material of Step 6, 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosan-1-yl]oxy}ethyl-1-amine into 1-{[1-( 26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-(3,4,5,6-tetrahydro-2H-pyran-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine was replaced to give the title compound (20 mg, yield: 32.6%). MS m / z (ESI): 1073.5 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.11 (s, 2H), 7.95 (s, 1H), 7.03 (s, 1H), 6.07 (s,2H), 4.11 (d, J = 7.6 Hz, 2H), 3.98-3.92 (m, 2H), 3.55-3.35 (m, 36H), 3.25-3.11(m, 3H), 2.92-2.82 (m, 4H), 2.70-2.65 (m, 2H), 2.41 (s, 2H), 2.36-2.31 (m, 2H),2.26 (t, J = 7.2 Hz, 2H), 1.97-1.93 (m, 2H), 1.86-1.65 (m, 8H), 1.44-1.34 (m,6H), 0.94 (t, J = 7.6 Hz, 3H).

[0366] Example 2-9: Preparation of N-{26-[4-({4-amino-2-butyl-7-[1-(2-methoxyethyl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-9)

[0367] [ka] Step 1: Preparation of tert-butyl ({26-[4-({4-amino-2-butyl-7-[1-(2-methoxyethyl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}amino)carboxylate The synthetic route of Example 24 was applied, and the raw material triethylene glycol monomethyl ether in step 3 was replaced with tert-butyl [(26-hydroxy-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl) amino] carboxylate, and the raw material tert-butyl 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino) thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl) methyl] hexahydropyridine-1-carboxylate was replaced with 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-[1-(2-methoxyethyl)hexahydropyridin-4-yl] thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to obtain the title compound (90 mg, yield: 24.7%). MS m / z (ESI): 980.6 [M+H] + .

[0368] Step 2: Preparation of 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-[1-(2-methoxyethyl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl ({26-[4-({4-amino-2-butyl-7-[1-(2-methoxyethyl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}amino)carboxylate to obtain the title compound (75 mg, yield: 93.7%). MS m / z (ESI): 880.5 [M+H] + .

[0369] Step 3: Preparation of N-{26-[4-({4-amino-2-butyl-7-[1-(2-methoxyethyl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Examples 2 and 3 was applied to convert the reactant material 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosan-1-yl]oxy}ethyl-1-amine in Step 6 into 1-{[1-( 26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-[1-(2-methoxyethyl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine was replaced to give the title compound (25 mg, yield: 28.1%). MS m / z (ESI): 1130.6 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.12 (s, 2H), 8.19 (s, 1H), 7.95 (s, 1H), 7.02 (s,1H), 6.09 (s, 2H), 4.12 (d, J = 7.2 Hz, 2H), 3.56-3.35 (m, 36H), 3.29-3.17 (m,5H), 3.05-2.80 (m, 7H), 2.60-2.50 (m, 6H), 2.30-2.12 (m, 5H), 2.02-1.92 (m,4H), 1.86-1.65 (m, 7H), 1.51-1.36 (m, 6H), 0.94 (t, J = 7.6 Hz, 3H).

[0370] Example 2-10: Preparation of N-[(1-{[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}-1,2,3-triazol-4-yl)methyl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-10)

[0371] [ka] Step 1: Preparation of 4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}[4-(hydroxymethyl)cyclohexyl]methanone The synthetic route of step 1 of Example 2-2 was applied, and 1-(4-aminobutyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine was replaced with 2-butyl-7-(hexahydropyridin-4-yl)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to obtain the title compound (0.27 g, yield: 86.8%). MS m / z (ESI): 568.4 [M+H] + .

[0372] Step 2: Preparation of 4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}carbonyl)cyclohexyl]hexahydropyridin-1-yl}carbonyl)hexahydropyridin-1-yl]methyl 4-methylbenzenesulfonate The synthetic route of step 2 of Example 2-2 was applied, and N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-(hydroxymethyl)cyclohexylcarboxamide was replaced with {4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}[4-(hydroxymethyl)cyclohexyl]methanone to obtain the title compound (0.20 g, yield: 51.2%). MS m / z (ESI): 722.3 [M+H]+ .

[0373] Step 3: Preparation of 4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}[4-(azidomethyl)cyclohexyl]methanone The synthetic route of step 3 of Example 2-2 was applied, and [4-({4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl}amino}carbonyl)cyclohexyl]4-methylbenzenesulfonate was replaced with [4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl 4-methylbenzenesulfonate to obtain the title compound (0.15 g, yield: 87.3%). MS m / z (ESI): 593.3 [M+H] + .

[0374] Step 4: Preparation of N-[(1-{[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}-1,2,3-triazol-4-yl)methyl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of step 4 of Example 2-2 was applied, and N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-(azidomethyl)cyclohexylcarboxamide was replaced with {4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}[4-(azidomethyl)cyclohexyl]methanone to obtain the title compound (56 mg, yield: 34.5%). MS m / z (ESI): 898.4 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.11 (s, 2H), 8.37 (d, J = 5.6 Hz, 1H), 7.92 (s, 1H),7.05 (s, 1H), 6.08 (s, 2H), 4.56-4.53 (m, 1H), 4.35-4.26 (m, 3H), 4.13 (d, J =7.6 Hz, 2H), 4.02 (s, 1H), 3.85-3.82 (m, 2H), 3.41 (s, 3H), 3.26-3.09 (m, 4H),2.88-2.62 (m, 5H), 2.36-2.25 (m, 3H), 2.15-1.96 (m, 5H), 1.84-1.77 (m, 4H),1.74-1.28 (m, 16H), 0.94 (t, J = 7.2 Hz, 3H).

[0375] Example 2-11: Preparation of N-(24-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-24-oxo-3,6,9,12,15,18,21-heptaoxotetracosan-1-yl)-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-11)

[0376] [ka] Step 1: Preparation of tert-butyl [(24-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-24-oxo-3,6,9,12,15,18,21-heptaoxatetracosan-1-yl)amino]carboxylate The synthetic route of step 1 of Example 2-2 was applied, and 1-(4-aminobutyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine was replaced with 2-butyl-7-(hexahydropyridin-4-yl)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine, and 4-(hydroxymethyl)cyclohexyl-1-carboxylic acid was replaced with 3-[(2,2-dimethyl-4-oxo-5-aza-3,8,11,14,17,20,23-heptaoxapentacosane-25-yl)oxy]propanoic acid to obtain the title compound (0.15 g, yield: 72.9%). MS m / z (ESI): 907.5 [M+H] + .

[0377] Step 2: Preparation of 1-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-3-[(20-amino-3,6,9,12,15,18-hexaoxaeicosan-1-yl)oxyl]propan-1-one The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl [(24-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-24-oxo-3,6,9,12,15,18,21-heptaoxatetracosan-1-yl)amino]carboxylate to obtain the title compound (0.12 g, yield: 86.5%). MS m / z (ESI): 807.5 [M+H] + .

[0378] Step 3: Preparation of N-(24-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-24-oxo-3,6,9,12,15,18,21-heptaoxatetracosan-1-yl)-6-[2-(methyldioxaylidene-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Example 2-3 was applied to convert the raw material of Step 6, 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosan-1-yl]oxy}ethyl-1-amine into 1-{4-[4-amino-2 -butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-3-[(20-amino-3,6,9,12,15,18-hexaoxaicosan-1-yl)oxy]propan-1-one to give the title compound (44 mg, yield: 27.1%). MS m / z (ESI): 1057.5 [M+H] + . 1 H NMR (400 MHz, CD 3 OD): δ 8.95 (s, 2H), 7.10 (s, 1H), 4.69-4.67 (m, 1H),4.24-4.14 (m, 3H), 3.95-3.91 (m, 2H), 3.80-3.73 (m, 2H), 3.65-3.48 (m, 27H),3.38-3.32 (m, 6H), 3.29-3.21 (m, 2H), 2.97-2.92 (m, 2H), 2.81-2.77 (m, 2H),2.66-2.55 (m, 3H), 2.39 (t, J = 7.6 Hz, 2H), 2.20-2.10 (m, 3H), 1.97-1.85 (m,4H), 1.82-1.60 (m, 2H), 1.59-1.46 (m, 6H), 1.02 (t, J = 7.6 Hz, 3H).

[0379] Example 2-12: Preparation of N-[(1-{[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}-1,2,3-triazacyclopentan-4-yl)methyl]-3-({27-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]-22-oxo-21-aza-3,6,9,12,15,18-hexaoxaheptacosane-26-yn-1-yl}oxa)propionamide (compound D-L'-12)

[0380] [ka] Step 1: Preparation of tert-butyl [(24-oxo-25-aza-3,6,9,12,15,18,21-heptaoxaoctacosan-27-yn-1-yl)amino]carboxylate The synthetic route of step 1 of Example 2-2 was applied, and 1-(4-aminobutyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine was replaced with propargylamine, and 4-(hydroxymethyl)cyclohexyl-1-carboxylic acid was replaced with 3-[(2,2-dimethyl-4-oxo-5-aza-3,8,11,14,17,20,23-heptaoxapentacosan-25-yl)oxy]propionic acid to obtain the title compound (0.16 g, yield: 72.7%). MS m / z (ESI): 535.3 [M+H] + .

[0381] Step 2: Preparation of 3-[(20-amino-3,6,9,12,15,18-hexaoxaeicosan-1-yl)oxy]-N-(prop-2-ynyl)propionamide The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl [(24-oxo-25-aza-3,6,9,12,15,18,21-heptaoxaoctacosan-27-yn-1-yl)amino]carboxylate to obtain the title compound (0.11 g, yield: 84.6%). MS m / z (ESI): 435.3 [M+H] + .

[0382] Step 3: Preparation of 3-({27-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]-22-oxo-21-aza-3,6,9,12,15,18-hexaoxaheptacosane-26-yn-1-yl}oxa)-N-(prop-2-ynyl)propionamide The synthetic route of Examples 2-3 was applied, and the raw material 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosane-1-yl]oxy}ethyl-1-amine in step 6 was replaced with 3-[(20-amino-3,6,9,12,15,18-hexaoxaeicosan-1-yl)oxy]-N-(prop-2-ynyl)propanamide to obtain the title compound (82 mg, yield: 46.2%). MS m / z (ESI): 685.3 [M+H] + . Step 4: Preparation of N-[(1-{[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}-1,2,3-triazacyclopentan-4-yl)methyl]-3-({27-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]-22-oxo-21-aza-3,6,9,12,15,18-hexaoxaheptacosane-26-yn-1-yl}oxa)propionamide

[0383] By applying the synthetic route of Step 4 of Example 2-2, N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-(azidomethyl)cyclohexylcarboxamide was converted to {4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}[4-(azidomethyl)cyclohexyl ]methanone and 6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]-N-(prop-1-ynyl-3-yl)hex-5-ynylamide was replaced with 3-({27-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]-22-oxo-21-aza-3,6,9,12,15,18-hexaoxaheptacosane-26-yn-1-yl}oxy)-N-(prop-2-ynyl)propionamide to give the title compound (28 mg, yield: 45.1%). MS m / z (ESI): 1277.6 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.12 (s, 2H), 8.35-8.32 (m, 1H), 7.97-7.94 (m, 1H),7.90 (s, 1H), 7.04 (s, 1H), 6.08 (s, 2H), 4.56-4.53 (m, 1H), 4.37-4.26 (m, 4H),4.15-4.11 (m, 2H), 4.03-4.01 (m, 1H), 3.85-3.81 (m, 2H), 3.66-3.39 (m, 38H),3.26-3.15 (m, 4H), 2.92-2.83 (m, 2H), 2.80-2.56 (m, 4H), 2.41-2.22 (m, 4H),2.10-2.06 (m, 4H), 1.82-1.72 (m, 4H), 1.53- 1.37 (m, 11H), 0.94 (t, J = 7.6 Hz,3H).

[0384] Example 2-13: Preparation of N-{2-[(3-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-3-oxopropyl)oxy]ethyl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-13)

[0385] [ka] Step 1: Preparation of tert-butyl ({2-[(3-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-3-oxopropyl)oxy]ethyl}amino)carboxylate The synthetic route of step 1 of Example 2-2 was applied, and 1-(4-aminobutyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine was replaced with 2-butyl-7-(hexahydropyridin-4-yl)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine, and 4-(hydroxymethyl)cyclohexyl-1-formic acid was replaced with 3-[(2,2-dimethyl-4-oxo-5-aza-3-oxaheptan-7-yl)oxy]propanoic acid to obtain the title compound (0.13 g, yield: 89.1%). MS m / z (ESI): 643.4 [M+H] + .

[0386] Step 2: Preparation of 1-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-3-[(2-aminoethyl)oxa]propan-1-one The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl ({2-[(3-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-3-oxopropyl)oxy]ethyl}amino)carboxylate to obtain the title compound (0.12 g, yield: 93.7%). MS m / z (ESI): 543.3 [M+H] + .

[0387] Step 3: Preparation of N-{2-[(3-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-3-oxopropyl)oxa]ethyl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Example 2-3 was applied, and the raw material 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18.21-heptaoxatricosane-1-yl]oxy}ethyl-1-amine in step 6 was replaced with 1-{4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}-3-[(2-aminoethyl)oxy]propan-1-one to obtain the title compound (43 mg, yield: 12.7%). MS m / z (ESI): 793.3 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6): δ 9.11 (s, 2H), 8.47 (s, 2H), 7.95-7.91 (m, 1H), 7.26(s, 1H), 4.56-4.52 (m, 1H), 4.24-4.19 (m, 2H), 4.03-3.99 (m, 1H), 3.87-3.82 (m,2H), 3.64 (t, J = 6.4 Hz, 2H), 3.43-3.39 (m, 6H), 3.34 -3.30 (m, 1H), 3.23-3.17(m, 4H), 3.16-3.12 (m, 1H), 2.96-2.90 (m, 2H), 2.71-2.59 (m, 3H), 2.54 (t, J =7.2 Hz, 2H), 2.30-2.26 (m, 2H), 2.06 (d, J = 7.6 Hz, 3H), 1.86-1.75 (m, 4H),1.69-1.58 (m, 1H), 1.50-1.40 (m, 6H), 0.95 (t, J = 7.2 Hz, 3H).

[0388] Example 2-14: Preparation of N-[(1-{[4-({4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}-1,2,3-triazacyclopent-4-yl)methyl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hexan-5-ynylamide (compound D-L'-14)

[0389] [ka] Step 1: Preparation of 4-[(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}[4-(hydroxymethyl)cyclohexyl]methanone The synthetic route of step 1 of Example 2-2 was applied, and 1-(4-aminobutyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine was replaced with 2-butyl-7-methyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (0.15 g, yield: 81.3%). MS m / z (ESI): 498.3 [M+H] + .

[0390] Step 2: Preparation of 4-({4-[(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl 4-methylbenzenesulfonate The synthetic route of step 2 of Example 2-2 was applied, and N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-(hydroxymethyl)cyclohexylcarboxamide was replaced with {4-[(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}[4-(hydroxymethyl)cyclohexyl]methanone to obtain the title compound (0.1 g, yield: 68.3%). MS m / z (ESI): 652.3 [M+H] + .

[0391] Step 3: Preparation of 4-[(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}[4-(azidomethyl)cyclohexyl]methanone The synthetic route of step 3 of Example 2-2 was applied, and [4-({4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl}amino}carbonyl)cyclohexyl]methyl 4-methylbenzenesulfonate was replaced with [4-({4-[(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl 4-methylbenzenesulfonate to obtain the title compound (94 mg, yield: 91.4%). MS m / z (ESI): 523.3 [M+H] + .

[0392] Step 4: Preparation of N-[(1-{[4-({4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}-1,2,3-triazolan-4-yl)methyl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of step 2 of Example 2-2 was applied, and N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-(azidomethyl)cyclohexylcarboxamide was replaced with {4-[(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}[4-(azidomethyl)cyclohexyl]methanone to obtain the title compound (12 mg, yield: 21.4%). MS m / z (ESI): 828.4 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6): δ 9.11 (s, 2H), 8.44-8.40 (m, 3H), 7.90 (s, 1H), 7.24(s, 1H), 4.52-4.16 (m, 7H), 3.94-3.91 (m, 1H), 3.00-2.83 (m, 3H), 2.68-2.50 (m,6H), 2.41-2.26 (m, 5H), 2.10-2.06 (m, 2H), 1.85-1.23(m, 19H), 0.95 (t, J = 7.6Hz, 3H).

[0393] Example 2-15: Preparation of N-(26-{4-[(4-amino-2-butylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-15)

[0394] [ka] Step 1: Preparation of tert-butyl [(26-{4-[(4-amino-2-butylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)amino]carboxylate The synthetic route of Example 24 was applied, and the raw material triethylene glycol monomethyl ether in step 3 was replaced with tert-butyl [(26-hydroxy-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl) amino] carboxylate, and the raw material 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino) thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl) methyl] hexahydropyridine-1-carboxylate was replaced with 2-butyl-1-(piperidin-4-ylmethyl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine to obtain the title compound (45 mg, yield: 39.6%). MS m / z (ESI): 839.5 [M+H] + .

[0395] Step 2: Preparation of 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butylthieno[3,2-b]imidazo[4,5-d]pyridin-4-amine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl [(26-{4-[(4-amino-2-butylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)amino]carboxylate to obtain the title compound (33 mg, yield: 84.5%). MS m / z (ESI): 739.5 [M+H] + .

[0396] Step 3: Preparation of N-(26-{4-[(4-amino-2-butylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridin-1-yl}-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Examples 2-3 was applied, and the reactant 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosane-1-yl]oxy}ethyl-1-amine in step 6 was replaced with 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butylthieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to obtain the title compound (12 mg, yield: 27.3%). MS m / z (ESI): 989.5 [M+H] + . 1 H NMR (400 MHz, CD 3 OD) δ: 8.96 (s, 2H), 8.02 (d, J = 5.6 Hz, 1H), 7.49 (d, J =5.6 Hz, 1H), 4.39 (d, J = 7.6 Hz, 2H), 3.83-3.80 (m, 2H), 3.73-3.46 (m,30H),3.38-3.32 (m, 8H), 2.99 (t, J = 7.6 Hz, 4H), 2.59 (t, J = 7.2 Hz, 2H), 2.40 (t,J = 7.2 Hz, 3H), 2.02- 1.74 (m, 8H), 1.61-1.48 (m, 3H), 1.04 (t, J = 7.6 Hz,3H).

[0397] Example 2-16: Preparation of N-[26-(4-{[4-amino-2-(2-methoxyethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-16)

[0398] [ka] Step 1: Preparation of tert-butyl {[26-(4-{[4-amino-2-(2-methoxyethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]amino}carboxylate The synthetic route of Example 24 was applied, and the raw material triethylene glycol monomethyl ether in step 3 was replaced with tert-butyl [(26-hydroxy-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl) amino] carboxylate, and the raw material 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino) thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl) methyl] hexahydropyridine-1-carboxylate was replaced with 1-(hexahydropyridin-4-ylmethyl)-2-(2-methoxyethyl) thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to obtain the title compound (52 mg, yield: 45.2%). MS m / z (ESI): 841.5 [M+H] + .

[0399] Step 2: Preparation of 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-(2-methoxyethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl {[26-(4-{[4-amino-2-(2-methoxyethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]amino}carboxylate to obtain the title compound (39 mg, yield: 86.5%). MS m / z (ESI): 741.5 [M+H] + .

[0400] Step 3: Preparation of N-[26-(4-{[4-amino-2-(2-methoxyethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Example 2-3 was applied, and the reactant 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosane-1-yl]oxy}ethyl-1-amine in step 6 was replaced with 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-(2-methoxyethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to obtain the title compound (20 mg, yield: 38.5%). MS m / z (ESI): 991.5 [M+H] + . 1 H NMR (400 MHz, CD 3OD): δ 8.95 (s, 2H), 7.68 (d, J = 5.6 Hz, 1H), 7.30 (d, J =5.6 Hz, 1H), 4.36-4.32 (m, 2H), 3.94-3.89 (m, 2H), 3.73-3.68 (m, 2H), 3.65-3.48(m, 32H), 3.42-3.35 (m, 8H), 3.23 (t, J = 6.4 Hz, 2H), 2.94 (s, 2H), 2.62-2.35(m, 6H), 2.24-2.10 (m, 1H), 1.95-1.90 (m, 2H), 1.81-1.57 (m, 4H).

[0401] Example 2-17: Preparation of N-[26-(4-{[4-amino-2-butyl-7-(1-methylhexahydropyridin-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-17)

[0402] [ka] Step 1: Preparation of tert-butyl {[26-(4-{[4-amino-2-butyl-7-(1-methylhexahydropyridin-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]amino}carboxylate The synthetic route of Example 24 was applied, and the raw material triethylene glycol monomethyl ether in step 3 was replaced with tert-butyl [(26-hydroxy-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl) amino] carboxylate, and the raw material 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino) thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl) methyl] hexahydropyridine-1-carboxylate was replaced with 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-(1-methylhexahydropyridin-4-yl) thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to obtain the title compound (52 mg, yield: 54.3%). MS m / z (ESI): 936.6 [M+H] + .

[0403] Step 2: Preparation of 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-(1-methylhexahydropyridin-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl {[26-(4-{[4-amino-2-butyl-7-(1-methylhexahydropyridin-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]amino}carboxylate to obtain the title compound (39 mg, yield: 84.5%). MS m / z (ESI): 836.5 [M+H] + .

[0404] Step 3: Preparation of N-[26-(4-{[4-amino-2-butyl-7-(1-methylhexahydropyridin-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl]methyl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Example 2-3 was applied to convert the raw material of Step 6, 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosan-1-yl]oxy}ethyl-1-amine to 1-{[ 1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-(1-methylhexahydropyridin-4-yl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine was substituted to give the title compound (23 mg, yield: 46.1%). MS m / z (ESI): 1086.6 [M+H] + . 1 H NMR (400 MHz, CD 3 OD): δ 8.95 (s, 2H), 7.11 (s, 1H), 4.28 (d, J = 7.6 Hz, 2H),3.75-3.71 (m, 2H), 3.65-3.49 (m, 30H), 3.43-3.33 (m, 8H), 3.18-3.15 (m, 1H),3.05-2.91 (m, 4H), 2.80-2.75 (m, 2H), 2.68 (s, 3H), 2.62-2.52 (m, 4H), 2.39 (t,J = 7.6 Hz, 2H), 2.31-2.11 (m, 4H), 2.05-1.86 (m, 6H), 1.79-1.65 (m 4H),1.55-1.48 (m, 2H), 1.03 (t, J = 7.6 Hz, 3H).

[0405] Example 2-18: Preparation of N-{26-[4-({4-amino-2-butyl-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-18)

[0406] [ka] Step 1: Preparation of tert-butyl ({26-[4-({4-amino-2-butyl-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}amino)carboxylate The synthetic route of Example 24 was applied, and the raw material triethylene glycol monomethyl ether in step 3 was converted to tert-butyl [(26-hydroxy-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)amino]carboxylate, and the raw material tert-butyl 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino)thieno[3,2-b] Imidazo[4,5-d]pyridin-1-yl)methyl]hexahydropyridine-1-carboxylate was replaced with 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to give the title compound (28 mg, yield: 61.3%). MS m / z (ESI): 1068.7 [M+H] + .

[0407] Step 2: Preparation of 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl ({26-[4-({4-amino-2-butyl-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octoxahexacosan-1-yl}amino)carboxylate to obtain the title compound (22 mg, yield: 86.5%). MS m / z (ESI): 968.7 [M+H] + .

[0408] Step 3: Preparation of N-{26-[4-({4-amino-2-butyl-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Example 2-3 was applied to convert the raw material of Step 6, 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosan-1-yl]oxy}ethyl-1-amine into 1-{[1-(26-amino -3,6,9,12,15,18,21,24-Octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-[1-(2,5,8-trioxadecan-10-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to give the title compound (15 mg, yield: 53.5%). MS m / z (ESI): 1218.7 [M+H] + . 1 H NMR (400 MHz, CD 3 OD): δ 8.96 (s, 2H), 7.06 (s,1H), 4.21 (d, J = 7.6 Hz, 2H),3.73-3.50 (m, 43H), 3.40-3.33 (m, 8H), 3.20-3.16 (m, 2H), 3.10-2.90 (m, 5H),2.75-2.70 (m, 2H), 2.65-2.55 (m, 4H), 2.42-2.32 (m, 4H), 2.15-1.85 (m, 10H),1.65-1.45 (m, 6H), 1.02 (t, J = 7.6 Hz, 3H).

[0409] Example 2-19: Preparation of N-{26-[4-({4-amino-2-butyl-7-[1-(2,5,8,11,14-pentaoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octoxahexacosan-1-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-19)

[0410] [ka] Step 1: Preparation of tert-butyl ({26-[4-({4-amino-2-butyl-7-[1-(2,5,8,11,14-pentaoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}amino)carboxylate Applying the synthetic route of Example 24, the raw material triethylene glycol monomethyl ether in step 3 was replaced with tert-butyl [(26-hydroxy-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)amino]carboxylate, and the raw material 4-[(2-butyl-7-(hexahydropyridin-4-yl)-4-(tert-butylamino)thieno[3,2-b]imidazo[4, 5-d]pyridin-1-yl)methyl]hexahydropyridine-1-carboxylate was replaced with 2-butyl-1-(hexahydropyridin-4-ylmethyl)-7-[1-(2,5,8,11,14-pentaoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to give the title compound (45 mg, yield: 56.3%). MS m / z (ESI): 1156.8 [M+H] + .

[0411] Step 2: Preparation of 1-{[1-(26-amino-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-[1-(2,5,8,11,14-pentaoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl ({26-[4-({4-amino-2-butyl-7-[1-(2,5,8,11,14-pentaoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosane-1-yl}amino)carboxylate to obtain the title compound (35 mg, yield: 85.3%). MS m / z (ESI): 1056.7 [M+H] + .

[0412] Step 3: Preparation of N-{26-[4-({4-amino-2-butyl-7-[1-(2,5,8,11,14-pentaoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-1-yl}methyl)hexahydropyridin-1-yl]-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Examples 2 and 3 was applied to convert the raw material of Step 6, 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosan-1-yl]oxy}ethyl-1-amine into 1-{[1-(26-amino-3,6, 9,12,15,18,21,24-Octaoxahexacosan-1-yl)hexahydropyridin-4-yl]methyl}-2-butyl-7-[1-(2,5,8,11,14-pentaoxahexadecan-16-yl)hexahydropyridin-4-yl]thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine to give the title compound (22 mg, yield: 51.2%). MS m / z (ESI): 1306.7 [M+H] + . 1 H NMR (400 MHz, CD 3 OD): δ 8.96 (s, 2H), 7.07 (s, 1H), 4.23 (d, J = 7.6 Hz, 2H),3.73-3.69 (m, 2H), 3.67-3.48 (m, 40H), 3.38-3.32 (m, 10H), 3.23-3.20 (m, 2H),3.15-2.99 (m, 4H), 2.98-2.92 (m, 2H), 2.72-2.62 (m, 4H), 2.58 (t, J = 7.2 Hz,2H), 2.49-2.35 (m, 4H), 2.22-1.84 (m, 12H), 1.69-1.46 (m, 7H), 1.32-1.15(m,3H), 1.02 (t, J = 7.6 Hz, 3H).

[0413] Example 2-20: Preparation of N-[2-({[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}oxy)ethyl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-20)

[0414] [ka] Step 1: Preparation of benzyl 4-(hydroxymethyl)cyclohexane-1-carboxylate 4-(Hydroxymethyl)cyclohexane-1-carboxylic acid (3 g, 19.0 mmol), potassium carbonate (5.25 g, 38.0 mmol) and tetrabutylammonium iodide (70.0 mg, 0.19 mmol) were added to N,N-dimethylformamide (20 mL) in sequence, cooled in an ice-water bath, bromotoluene (3.9 g, 22.8 mmol) was slowly added to the reaction solution at 0° C., slowly heated to room temperature and stirred for 24 hours. The reaction solution was poured into water (150 mL), extracted three times with ethyl acetate (75 mL), the organic phases were combined, washed three times with saturated aqueous sodium bicarbonate solution (30 mL), washed three times with saturated saline solution (30 mL), the organic phase was dehydrated over anhydrous sodium sulfate, concentrated, and the resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate: petroleum ether = 5: 1, volume ratio) to obtain the title compound (4.3 g, yield: 91.7%). MS m / z (ESI): 249.2 [M+H] + .

[0415] Step 2: Preparation of benzyl 4-(4-oxo-2,5-dioxahept-1-yl)cyclohexane-1-carboxylate Benzyl 4-(hydroxymethyl)cyclohexane-1-carboxylate (1g, 4.03mmol) and rhodium dipolyacetate (89.0mg, 0.2mmol) were added to dichloromethane (20mL) in sequence, cooled in an ice-water bath, ethyl diazoacetate (690.0mg, 6.04mmol) was slowly added dropwise to the reaction solution at 0°C, the reaction system was bubbled with nitrogen for 5 minutes, and the reaction was allowed to proceed at room temperature for 18 hours. The reaction solution was poured into water (60mL), extracted three times with ethyl acetate (30mL), the organic phases were combined, washed twice with saturated saline (15mL), the organic phase was dehydrated over anhydrous sodium sulfate, concentrated, and the resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate: petroleum ether = 5:1, volume ratio) to obtain the title compound (475.7mg, yield: 35.3%). MS m / z (ESI): 335.2 [M+H] + .

[0416] Step 3: Preparation of 4-(4-oxo-2,5-dioxahept-1-yl)cyclohexane-1-carboxylic acid Benzyl 4-(4-oxo-2,5-dioxahept-1-yl)cyclohexane-1-carboxylate (300 mg, 0.90 mmol) was dissolved in methanol (3 mL) and added together with 10% palladium on carbon (15.0 mg, 0.09 mmol), purged with hydrogen three times, the reaction was stirred at room temperature for 2 hours, suction filtered, the filter cake was washed with methanol (3 mL), and the filtrate was concentrated to give the title compound (190.0 mg, yield: 86.3%). MS m / z (ESI): 245.2 [M+H] + .

[0417] Step 4: Preparation of ethyl ({[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}oxy)acetate The synthetic route of Example 2-2 was applied, and the raw material 1-(4-aminobutyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 1 was replaced with 2-butyl-7-(hexahydropyridin-4-yl)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine, and 4-(hydroxymethyl)cyclohexyl-1-carboxylic acid was replaced with 4-(4-oxo-2,5-dioxahept-1-yl)cyclohexane-1-carboxylic acid to obtain the title compound (277 mg, yield: 98.3%). MS m / z (ESI): 654.4 [M+H] + .

[0418] Step 5: Preparation of 4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}(4-{[(2-hydroxyethyl)oxy]methyl}cyclohexyl)methanone Ethyl ({[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}oxy)acetate (270 mg, 0.41 mmol) was dissolved in a mixed solvent of tetrahydrofuran (4 mL) and absolute ethanol (6 mL), calcium chloride (64.4 mg, 0.58 mmol) and sodium borohydride (65.4 mg, 1.73 mmol) were added at room temperature, and the mixture was heated to 60° C. and stirred for 8 hours. The reaction mixture was poured into water (60 mL) and extracted three times with dichloromethane (45 mL). The organic phases were combined and washed three times with saturated brine (15 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 8:1, volume ratio) to obtain the title compound (150 mg, yield: 59.8%). MS m / z (ESI): 612.4 [M+H] + .

[0419] Step 6: Preparation of 2-({4-(4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}oxy)ethyl methanesulfonate The synthetic route of Example 2-2 was applied, and the raw material N-[4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl]-4-(hydroxymethyl)cyclohexylcarboxamide in step 2 was replaced with {4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}(4-{[(2-hydroxyethyl)oxy]methyl}cyclohexyl)methanone, and p-toluenesulfonyl chloride was replaced with methanesulfonyl chloride to obtain the title compound (140 mg, yield: 82.8%). MS m / z (ESI): 690.3 [M+H] + .

[0420] Step 7: Preparation of 4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}(4-{[(2-azidoethyl)oxy]methyl}cyclohexyl)methanone The synthetic route of embodiment 2-2 was applied, and the raw material [4-({4-(4-amino-2-butyl-7-methylthieno[3,2-b]imidazo[4,5-d]pyridin-1-yl)butyl}amino}carbonyl)cyclohexyl]4-methylbenzenesulfonate in step 3 was replaced with 2-({4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}oxy)ethyl methanesulfonate to obtain the title compound (119 mg, yield: 92.2%). MS m / z (ESI): 637.4 [M+H] + .

[0421] Step 8: Preparation of 4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}(4-{[(2-aminoethyl)oxyl]methyl}cyclohexyl)methanone {4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}(4-{[(2-azidoethyl)oxyl]methyl}cyclohexyl)methanone (119 mg, 0.19 mmol) was dissolved in tetrahydrofuran (3 mL) and ammonia (H 2 The mixture was dissolved in a mixed solvent of 1,2-dichloromethane (14 wt.% in 2O, 2 mL), triphenylphosphine (62.9 mg, 0.24 mmol) was added at room temperature, and the mixture was heated to 50° C. and stirred for 6 hours. After cooling to room temperature, the reaction system was concentrated to remove the solvent, and the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol=9:1, volume ratio) to obtain the title compound (101.8 mg, yield: 87.7%). MS m / z (ESI): 611.4 [M+H] + .

[0422] Step 9: Preparation of N-[2-({[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]methyl}oxy)ethyl]-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Examples 2-3 was applied, and the reactant 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosane-1-yl]oxy}ethyl-1-amine in step 6 was replaced with {4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}(4-{[(2-aminoethyl)oxy]methyl}cyclohexyl)methanone to obtain the title compound (20 mg, yield: 44.9%). MS m / z (ESI): 861.4 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.11 (s, 2H), 7.95-7.93 (m, 1H), 7.03 (s, 1H), 6.07(s, 2H), 4.50 (s, 1H), 4.13 (d, J = 7.6 Hz, 2H), 3.99 (s, 1H), 3.84-3.82 (m,2H), 3.41 (s, 3H), 3.40-3.38 (m, 2H), 3.24-3.15 (m, 5H), 2.88-2.82 (m, 2H),2.69-2.65 (m, 3H), 2.56 (d, J = 7.2 Hz, 2H), 2.33 (s, 1H), 2.26 (t, J = 7.2 Hz,2H), 2.05 (s, 4H), 1.84-1.74 (m, 4H), 1.63-1.38 (m, 16H), 1.23 (s, 1H), 0.94(t, J = 7.2 Hz, 3H).

[0423] Example 2-21: Preparation of N-{1-[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]-2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide (compound D-L'-21)

[0424] [ka] Step 1: Preparation of tert-butyl 4-(acetoxymethyl)cyclohexane-1-carboxylate 4-(Hydroxymethyl)cyclohexane-1-carboxylic acid (2g, 12.6mmol) was added to tert-butyl acetate (30mL), cooled in an ice-water bath, perchloric acid (0.13g, 1.26mmol) was slowly added to the reaction solution at 0°C, slowly warmed to room temperature, and stirred for 12 hours. The reaction solution was poured into water (100mL), extracted three times with dichloromethane (60mL), the organic phases were combined, washed twice with saturated sodium bicarbonate (30mL), washed three times with saturated saline (45mL), and the organic phase was dehydrated over anhydrous sodium sulfate and concentrated to obtain the title compound (3g, yield: 92.9%). MS m / z (ESI): 257.2 [M+H] + .

[0425] Step 2: Preparation of tert-butyl 4-(hydroxymethyl)cyclohexane-1-carboxylate tert-Butyl 4-(acetoxymethyl)cyclohexane-1-carboxylate (3 g, 11.7 mmol) was dissolved in a mixed solvent of methanol (30 mL) and water (6 mL), cooled in an ice-water bath, lithium hydroxide (1.4 g, 58.5 mmol) was slowly added to the reaction solution at 0° C., slowly warmed to room temperature, and stirred for 3 hours. The reaction solution was poured into water (100 mL), extracted three times with ethyl acetate (75 mL), the organic phases were combined, washed three times with saturated saline (30 mL), the organic phase was dehydrated with anhydrous sodium sulfate, concentrated, and the obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate: petroleum ether = 1:5, volume ratio) to obtain the title compound (1.2 g, yield: 47.9%). MS m / z (ESI): 215.2 [M+H] + .

[0426] Step 3: Preparation of tert-butyl 4-(32,32-dimethyl-30-oxo-29-aza-2,5,8,11,14,17,20,23,26,31-decaoxatritriacontan-1-yl)cyclohexane-1-carboxylate tert-Butyl 4-(hydroxymethyl)cyclohexane-1-carboxylate (0.22 g, 1.03 mmol) was dissolved in tetrahydrofuran (6 mL) and cooled in an ice-water bath. Potassium tert-butoxide (0.12 g, 1.03 mmol) was added to the reaction solution at 0°C; after stirring for 20 minutes, tert-butyl [(26-bromo-3,6,9,12,15,18,21,24-octaoxahexacosan-1-yl)amino]carboxylate (0.59 g, 1.03 mmol) in 2 mL of tetrahydrofuran was slowly added dropwise to the reaction solution, which was then slowly warmed to room temperature and stirred for 12 hours. The reaction mixture was poured into a saturated aqueous solution of ammonium chloride (50 mL) and extracted three times with dichloromethane (45 mL). The organic phases were combined and washed three times with saturated brine (30 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated to obtain the title compound (0.5 g, yield: 68.6%). MS m / z (ESI): 710.5 [M+H] + .

[0427] Step 4: Preparation of 4-(32,32-dimethyl-30-oxo-29-aza-2,5,8,11,14,17,20,23,26,31-decaoxatritriacontan-1-yl)cyclohexane-1-carboxylic acid tert-Butyl 4-(32,32-dimethyl-30-oxo-29-aza-2,5,8,11,14,17,20,23,26,31-decaoxatritriacontan-1-yl)cyclohexane-1-carboxylate (0.18 g, 0.25 mmol) was dissolved in a mixture of tetrahydrofuran (2 mL) and methanol (1 mL), cooled in an ice-water bath, and sodium hydroxide (0.6 g, 15.2 mmol) in an aqueous solution (3.8 mL) was slowly added to the reaction solution at 0° C., slowly heated to 60° C., and stirred for 12 hours. The reaction mixture was poured into water (30 mL) and extracted three times with dichloromethane (15 mL). The remaining aqueous phase was slowly adjusted to pH=7 using dilute hydrochloric acid (2N, 7.5 mL) and extracted three times with dichloromethane (30 mL). The organic phases were combined and washed three times with saturated saline (15 mL). The organic phase was dehydrated over anhydrous sodium sulfate and concentrated. The resulting residue was purified by high performance liquid chromatography to obtain the title compound (0.1 g, yield: 61.2%). MS m / z (ESI): 654.4 [M+H] + .

[0428] Step 5: Preparation of tert-butyl ({1-[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]-2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl}amino)carboxylate The synthetic route of Example 2-2 was applied, and the raw material 1-(4-aminobutyl)-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-4-amine in step 1 was replaced with 2-butyl-7-(hexahydropyridin-4-yl)-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-4-amine, and 4-(hydroxymethyl)cyclohexyl-1-carboxylic acid was replaced with 4-(32,32-dimethyl-30-oxo-29-aza-2,5,8,11,14,17,20,23,26,31-decaoxatritriacontan-1-yl)cyclohexane-1-carboxylic acid to obtain the title compound (37 mg, yield: 61.5%). MS m / z (ESI): 1063.6 [M+H] + .

[0429] Step 6: Preparation of {4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}[4-(28-amino-2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-1-yl)cyclohexyl]methanone The synthetic route of Example 1 was applied, and the raw material tert-butyl (4-(4-amino-2-butyl-7-methyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl)butyl)carbamate in step 9 was replaced with tert-butyl ({1-[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]-2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl}amino)carboxylate to obtain the title compound (32 mg, yield: 95.2%). MS m / z (ESI): 963.6 [M+H] + .

[0430] Step 7: Preparation of N-{1-[4-({4-[4-amino-2-butyl-1-(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}carbonyl)cyclohexyl]-2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl}-6-[2-(methyldioxo-λ6-thio)pyrimidin-5-yl]hex-5-ynylamide The synthetic route of Example 2-3 was applied to convert the raw material 2-{[23-(4-{2-butyl-4-(tert-butylamino)-1H-thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl}hexahydropyridin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosan-1-yl]oxy}ethyl-1-amine to {4-[4-amino-2-butyl-1 -(3,4,5,6-tetrahydro-2H-pyran-4-ylmethyl)thieno[3,2-b]imidazo[4,5-d]pyridin-7-yl]hexahydropyridin-1-yl}[4-(28-amino-2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-1-yl)cyclohexyl]methanone to give the title compound (23 mg, yield: 55.3%). MS m / z (ESI): 1213.6 [M+H] + . 1 H NMR (400 MHz, CD 3OD): δ 8.97 (s, 2H), 7.11 (s, 1H), 4.69-4.67 (m, 1H),4.25-4.10 (m, 3H), 3.94 (d, J = 11.2 Hz, 3H), 3.73-3.43 (m, 36H), 3.42-3.33 (m,6H), 3.26-3.21 (m, 2H), 3.01-2.90 (m, 2H), 2.75-2.70 (m, 2H), 2.58 (t, J = 7.2Hz, 2H), 2.40 (t, J = 7.2 Hz, 2H), 2.26-2.11 (m, 3H), 2.00-1.40 (m, 20H),1.32-1.22 (m 1H), 1.02 (t, J = 7.2 Hz, 3H).

[0431] III. Preparation of Immunostimulatory Antibody Conjugates (ISACs) Example 3-1: ISAC-1

[0432] [ka] The trastuzumab monoclonal antibody stock solution was adjusted to pH=7.24 with 1M disodium hydrogen phosphate solution, then diluted to 10mg / mL with phosphate buffered saline (PBS) of pH=7.2-7.4 to obtain trastuzumab monoclonal antibody buffer solution. DMSO was used as a solvent to prepare a 70mM SATA crosslinker solution. A 10-fold molar amount of trastuzumab monoclonal antibody SATA crosslinker solution was added to the trastuzumab monoclonal antibody buffer solution at room temperature, shaken to mix thoroughly, and then reacted at room temperature for 1 hour, then purified with a NAP-5 gel column, washed with phosphate buffered saline (PBS) of pH=7.2-7.4, and the filtrate was collected as the buffer solution of trastuzumab monoclonal antibody modified with SATA crosslinker.

[0433] 0.05M hydroxylamine and 2.5mM EDTA were added to the resulting buffer solution of trastuzumab monoclonal antibody modified with SATA crosslinker, shaken to mix thoroughly, and reacted at room temperature for 2 hours, then purified on a NAP-5 gel column and washed with phosphate buffered saline (PBS) containing 5mM EDTA at pH=7.2-7.4, and the filtrate was collected as the buffer solution of purified deacetylated SATA modified trastuzumab monoclonal antibody.

[0434] To the buffer solution of the purified deacetylated SATA-modified trastuzumab monoclonal antibody, the target compound of Example 2-1 in a DMSO solution (10 mM) of 10 times the molar amount of trastuzumab monoclonal antibody was added, shaken to mix thoroughly, and reacted at room temperature for 3 hours, then purified with a NAP-5 gel column, washed with phosphate buffered saline (PBS) at pH=7.2-7.4, and the filtrate was collected to obtain ISAC-1 (3.4 mg / mL, 3.2 mL) in PBS buffer, and stored at 4°C.

[0435] Example 3-2 The trastuzumab monoclonal antibody stock solution was adjusted to pH=7.6-7.7 with 1M disodium hydrogen phosphate solution, and then diluted to 10 mg / mL with phosphate buffered saline (PBS) at pH=7.6-7.7 to obtain a trastuzumab monoclonal antibody buffer solution. 0.1M EDTA and TCEP (5.5 equivalents, 10 mM) were added to the trastuzumab monoclonal antibody buffer solution in order, shaken to mix thoroughly, and reacted at room temperature for 2 hours. A 10-fold molar amount of the conjugate intermediate in a DMSO solution (10 mM) of the trastuzumab monoclonal antibody was added to the reaction system at room temperature, shaken to mix thoroughly, and reacted at room temperature for 2 hours. The mixture was then purified on a NAP-5 gel column, washed with a histidine hydrochloride buffer solution of pH=6.0-6.1, and the filtrate was collected to obtain ISAC-2, ISAC-3, ISAC-4, ISAC-5, ISAC-6, and ISAC-7 in PBS buffer, and stored at 4°C. IV. Drug / Antibody Ratios for Immunostimulatory Antibody Conjugates (ISACs): Determination of DAR Values The molecular weight of ISAC was determined by LC-MS, and the drug / antibody ratio DAR value was calculated. Chromatographic determination conditions: Liquid chromatography column: Thermo MAbPac RP 3.0*100mm; Mobile phase A: 0.1% FA / H 2 O; Mobile phase B: 0.1% FA / ACN; Flow rate: 0.25 ml / min; sample chamber temperature: 8°C; column temperature: 60°C; injection volume: 2 μl;

[0436] [Table 2] Mass spectrometry conditions: Mass spectrometry model: AB Sciex Triple TOF 5600+; GS1 35;GS2 35;CUR 30;TEM 350;ISVF 5500;DP 250;CE 10;Integration time 0.5s; m / z 600-4000, time bins-40 total. The mean DAR values ​​of the immunostimulatory antibody conjugates (ISACs) were calculated using CE-SDS and the results are shown in Table 5:

[0437] [Table 3]

[0438] Biological testing Experimental Example 1: Determination of the agonist activity of compounds on HEK-Blue hTLR7 cells Experimental steps: 1. HEK-Blue hTLR7 cells (Invivogen) were cultured in DMEM medium (Hyclone) containing 10% FBS heat-inactivated fetal bovine serum (Corning). On the day of detection, the state of the cells was observed under a microscope, the cells were collected and resuspended, the cell concentration was adjusted to 50 μL per well after counting, and the total amount of cells was 2 × 10 4 and the cells were seeded into 96-well plates.

[0439] 2. After the cells were attached to the wall overnight, 50 μL of test compounds were added to the well plate at different concentrations, with final concentrations of 100 μM, 33.3 μM, 11.11 μM, 3.70 μM, 1.23 μM, 0.41 μM, 0.14 μM, 0.05 μM, and 0 μM, and the final concentration of DMSO was 1%. The compounds were incubated at 37° C., CO 2 The mixture was incubated with the cells for 20 h in a 5% incubator.

[0440] 3. After incubation, observation was performed under a microscope to determine the maximum signal well, the state of cells with the highest and lowest compound concentrations, and whether the compound crystallized at high concentrations. 10 μL of cell culture supernatant was taken and transferred to a new 96-well transparent plate, and 90 μL of QUANTI-Blue (Invivogen) detection reagent was added to each well and incubated at 37°C for 3 h. OD 620 The results were read using a multi-function microplate reader (BMG LABTECH).

[0441] 4. EC 50 was calculated by fitting log(agonist) vs. response-variable slope with Graphpad Prism software, and Emax was the OD at which the activation effect of the test compound was maximal. 620 It was decided.

[0442] Test Results: The activation activity of the compounds against HEK-Blue hTLR7 cells was determined according to the above method, and the activity of the compounds was expressed as EC 50 and Emax, and the results are shown in Table 1.

[0443] [Table 4]

[0444] These results showed that compounds 1, 2, 3, 4, 5, 6, 8, 15, 16, 17, 25, 26, 27, 28, 32, 33, 34, 38, 40, 41, 42, 55, 56, and 59 had strong activating effects on human TLR7.

[0445] Experimental Example 2: Determination of the agonist activity of compounds on HEK-Blue hTLR8 cells Experimental steps: 1. HEK-Blue hTLR8 cells (Invivogen) were cultured in DMEM medium (Hyclone) containing 10% FBS heat-inactivated fetal bovine serum (Corning). On the day of detection, the state of the cells was observed under a microscope, the cells were collected and resuspended, the cell concentration was adjusted to 50 μL per well after counting, and the total amount of cells was 2 × 10 4 and the cells were seeded into 96-well plates.

[0446] 2. After the cells had attached to the wall overnight, 50 μL of test compounds were added to the well plate at different concentrations, resulting in final concentrations of 100 μM, 25 μM, 6.25 μM, 1.56 μM, 0.39 μM, 0.10 μM, 0.02 μM, 0.006 μM, 0 μM, or 100 μM, 33.3 μM, 11.11 μM, 3.70 μM, 1.23 μM, 0.41 μM, 0.14 μM, 0.05 μM, 0 μM, and a final concentration of DMSO of 1%. The compounds were incubated at 37° C., CO 2 The mixture was incubated with the cells for 20 hours in a 5% incubator.

[0447] 3. After incubation, observation was performed under a microscope to determine the maximum signal well, the state of cells with the highest and lowest concentrations of compound, and whether the compound crystallized at high concentrations. 10 μL of cell culture supernatant was transferred to a new 96-well transparent plate, and 90 μL of QUANTI-Blue (Invivogen) detection reagent was added to each well and incubated at 37°C for 3 hours, and the OD 620 The results were read using a multi-function microplate reader (BMG LABTECH).

[0448] 4. EC50 was calculated by fitting log(agonist) vs. response-variable slope with Graphpad Prism software, and Emax was the OD at which the activation effect of the test compound was maximal. 620 It was decided.

[0449] Test Results: The activation activity of the compounds against HEK-Blue hTLR8 cells was determined according to the method described above, and the activity of the compounds was expressed as EC 50 and Emax, and the results are shown in Table 2.

[0450] [Table 5]

[0451] These results demonstrated that compounds 1, 2, 3, 6, 27 and 33 had strong activating effects on human TLR8.

[0452] Experimental Example 3: Determining the ability of immunostimulatory antibody conjugates (ISACs) to stimulate peripheral blood mononuclear cells (PBMCs) to secrete TNF-α in the presence of tumor cells Experimental steps: 1. Logarithmic growth phase HCC1954 tumor cells expressing HER2 (Kangnuotai) were harvested and washed twice with PBS. The cells were then resuspended in RPMI1640 + 10% heat-inactivated FBS complete medium to a cell density of 2 × 10 5 The HCC1954 tumor cells were cultured in a 96-well plate (Corning) at 1 × 10 per well, adjusted to a concentration of 1 × 10 per mL. 4 The cells were inoculated.

[0453] 2. PBMCs (Sai ​​Li Biology) were resuscitated and resuspended in RPMI1640 + 10% heat-inactivated FBS complete medium to a density of 6 × 10 5 The tumor cells were seeded in a 96-well plate, and the tumor cells were added to the wells at 3 × 10 cells / mL. 4 PBMCs were added.

[0454] 3.4.6 8.4 nM of Trastuzumab monoclonal antibody and immune stimulatory antibody conjugate ISAC-1, and 55.6 nM of immune stimulatory antibody conjugates ISAC-2, ISAC-3, ISAC-4, ISAC-5, ISAC-6 and ISAC-7, prepared by dilution in 100 μL of RPMI 1640 + 10% heat inactivated FBS complete medium, were added to wells respectively, and empty wells were added with 100 μL of RPMI 1640 + 10% heat inactivated FBS complete medium.

[0455] 5.CO 2 After 48 hours of incubation in a 5% CO2 incubator at 37°C, 50 μL of supernatant was taken from each well for detection. Primary and HRP-labeled secondary antibodies were incubated, and color development was performed according to the steps of the Human TNF-α ELISA Detection Kit (Invitrogen), and the absorbance was read at 450 nm. The concentration of TNF-α was calculated according to the standard curve using Graphpad Prism software.

[0456] Test Results: The ability of trastuzumab monoclonal antibody and each immunostimulatory antibody conjugate (ISAC) to stimulate peripheral blood mononuclear cells (PBMCs) to secrete TNF-α in the presence of HCC1954 tumor cells was determined according to the methods described above, and the results are shown in Table 3:

[0457] [Table 6]

[0458] These results demonstrated that the immunostimulatory antibody conjugates ISAC-1, ISAC-2, ISAC-3, ISAC-4, ISAC-5, ISAC-6 and ISAC-7 stimulated greater TNF-α secretion than trastuzumab monoclonal antibody at comparable concentrations in an in vitro co-incubation system of HCC1954 tumor cells and PBMCs.

[0459] Experimental Example 4: Efficacy evaluation of ISAC in a SCID-beige mouse xenograft tumor model Experimental steps: 1.5×10 6 HCC1954 cells (mixed with Matrigel in an equal volume at 1:1 ratio and suspended in 0.1 mL of PBS) were subcutaneously inoculated into the right armpit of each SCID beige mouse (Weitong Lihua Co., Ltd.).

[0460] 2. The tumor grows to about 150 mm 3 When the tumor volume of each mouse was 100-1500 μg / kg, the mice with irregular tumor volume, too small tumor volume, and too large tumor volume were excluded, and the remaining mice were randomly divided into groups according to tumor volume and animal body weight, and were administered with saline (vehicle control), trastuzumab monoclonal antibody (9.95 mg / kg), and immune stimulatory antibody conjugate ISAC-1 (10 mg / kg, the same molar concentration as trastuzumab monoclonal antibody) by single intraperitoneal injection (ip), respectively, to observe the efficacy of the test compound on the tumor-bearing mouse model and the animal tolerance of the test compound.

[0461] 3. During the experiment, the mice were weighed twice a week, tumor volumes were measured, and the data were recorded.

[0462] 4. Data statistics, calculation formula for tumor volume (V): V = 1 / 2 × a × b 2 (wherein a and b represent length and width, respectively). The efficacy of an anticancer drug was evaluated in terms of tumor growth inhibition rate (TGI) (%), calculated using the following formula: TGI (%) (tumor volume) =[1-(T Vt -T V0 ) / (C Vt -C V0 )]×100% (where, T V0 represents the average tumor volume of the test compound group at the time of grouping and administration, and T Vt represents the average tumor volume of the test compound group on day t after administration, and C V0 represents the mean tumor volume of the vehicle group at the time of grouping and administration, and C Vt (TGI (%)) indicates the average tumor volume in the vehicle group on day t after administration.(tumor volume) =100%-(T Vt -T V0 ) / T V0 When the tumor shrinks from its initial volume, i.e., V t <V 0 If the tumor disappeared completely, this was defined as a complete tumor regression (CR).

[0463] Test Results: FIG. 1 shows the time course curves of tumor volume for each group in Experimental Example 4, which shows the in vivo efficacy of each test substance against a SCID-beige mouse xenograft tumor model subcutaneously inoculated with HCC1954 human breast cancer cells. 21 days after administration, the immunostimulatory antibody conjugate ISAC-1 was able to significantly inhibit tumor growth, with a tumor volume TGI of 180.25%, P<0.001, compared with the vehicle control group, while the trastuzumab monoclonal antibody showed no obvious effect (FIG. 1, Table 4). These results showed that a single intraperitoneal (ip) injection of the immunostimulatory antibody conjugate ISAC-1 showed a highly significant antitumor effect, clearly superior to that of the trastuzumab monoclonal antibody, in a SCID-beige mouse xenograft tumor model subcutaneously inoculated with HCC1954 human breast cancer cells.

[0464] [Table 7]

[0465] The above examples are not intended to limit the solutions of the present application in any way. In addition to those described herein, various modifications of the present invention will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited in this application, including all patents, patent applications, journal articles, books, and other publications, is hereby incorporated by reference in its entirety.

Claims

1. Formula (I): 【Chemistry 1】 (In the formula, X １ is selected from the group consisting of N and C; X ２ is selected from the group consisting of N and C; X １ and X ２ is selected from N; X ３ is selected from the group consisting of O, S and N; X ４ is O, S, N and CR ４ selected from the group consisting of: R １ is C １～６ Alkyl and -C １～６ Alkylene -O-C １～６ selected from the group consisting of alkyl; R ２ is selected from hydrogen or a group of formula -L １ -L ２ -L ３ -L ４ having L １ is a covalent bond and C １～６ alkylene; L ２ is a covalent bond, C ３～１０ Cycloalkyl, 3- to 12-membered heterocyclyl, C ６～１０ aryl and 5-10 membered heteroaryl, optionally wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C １～６ Alkyl, C １～６ Alkoxy and -C １～６ Alkylene-NH ２ each independently substituted with one or more groups independently selected from the group consisting of: L ３ is a covalent bond, -O-, -S-, -C(O)-, -O-C(O)-, -C(O)-O-, -O-C(O)-O-, -NR ５ -, -C(O)-NR ５ --, --NR ５ -C(O)-, -OC(O)-NR ５ --, --NR ５ -C(O)-O-, -NR ５ -C(O)-NR ５ -, -S(O) ｍ -NR ５ --, --NR ５ -S (O) ｍ - and -NR ５ -S (O) ｍ -NR ５ - selected from the group consisting of; L ４ is hydrogen, C １～６ Alkyl, -(O-CH ２ CH ２ ) ｎ -O-C １～６ Alkyl and -C １～６ Alkylene-(O-CH ２ CH ２ ) ｎ -O-C １～６ alkyl, optionally each alkyl being selected from the group consisting of hydrogen, halogen, hydroxyl, -NH ２ , -NH-C(O)-OC １～６ Alkyl, C １～６ Alkoxy, carboxyl and -C(O)-O-C １～６ independently substituted with one or more groups independently selected from the group consisting of alkyl; R ３ is hydrogen or -L ５ -L ６ -L ７ -L ８ having the formula: L ５ is a covalent bond and C １～６ alkylene; L ６ is a covalent bond, C ３～１０ Cycloalkyl, 3- to 12-membered heterocyclyl, C ６～１０ aryl and 5-10 membered heteroaryl, optionally wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C １～６ Alkyl, C １～６ Alkoxy and -C １～６ Alkylene-NH ２ each independently substituted with one or more groups independently selected from the group consisting of: L ７ is a covalent bond, -O-, -S-, -C(O)-, -O-C(O)-, -C(O)-O-, -O-C(O)-O-, -NR ６ -, -C(O)-NR ６ --, --NR ６ -C(O)-, -OC(O)-NR ６ --, --NR ６ -C(O)-O-, -NR ６ -C(O)-NR ６ -, -S(O) ｐ -NR ６ --, --NR ６ -S (O) ｐ - and -NR ６ -S (O) ｐ -NR ６ - selected from the group consisting of; L ８ is hydrogen, C １～６ Alkyl, -(O-CH ２ CH ２ ) ｑ -O-C １～６ Alkyl and -C １～６ Alkylene-(O-CH ２ CH ２ ) ｑ -O-C １～６ alkyl, optionally each alkyl being selected from the group consisting of hydrogen, halogen, hydroxyl, -NH ２ , -NH-C(O)-OC １～６ Alkyl, C １～６ Alkoxy, carboxyl and -C(O)-O-C １～６ independently substituted with one or more groups independently selected from the group consisting of alkyl; R ４ is hydrogen, halogen, cyano, hydroxyl, C １～６ Alkyl, C １～６ Haloalkyl, C １～６ Alkoxy and C ３～１０ cycloalkyl; Each R ５ is hydrogen and C １～６ independently selected from the group consisting of alkyl; Each R ６ is hydrogen and C １～６ independently selected from the group consisting of alkyl; each m is independently 1 or 2; Each n is independently selected from the group consisting of integers 1 to 25; each p is independently 1 or 2; and each q is independently selected from the group consisting of integers 1 to 25, or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

2. R １ is C １～４ Alkyl and -C １～４ Alkylene -O-C １～４ selected from the group consisting of alkyl; Preferably, R １ is C １～４ Alkyl and -C １～２ Alkylene -O-C １～２ selected from the group consisting of alkyl; Preferably, R １ is selected from the group consisting of n-butyl, 2-methoxyethyl, or ethoxymethyl; Preferably, R １ is n-butyl; 10. The compound of claim 1 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

3. R ２ is hydrogen, or R ２ Is, -L １ -L ２ -L ３ -L ４ Formula (In the formula, L １ represents a covalent bond and, for example, -CH ２ --, --CH ２ -CH ２ --, --CH ２ -CH ２ -CH ２ - or -CH ２ -CH ２ -CH ２ -CH ２ - and other C １～６ alkylene; preferably, L １ is a covalent bond or -CH ２ - and L ２ is a covalent bond, C ３～１０ Cycloalkyl (e.g., C such as piperidinyl) ３～６ Cycloalkyl or C ５～６ cycloalkyl), 3-12 membered heterocyclyl (e.g., 3-6 membered saturated or unsaturated nitrogen-containing monoheterocyclyl; e.g., 3-6 membered saturated or unsaturated oxygen-containing monoheterocyclyl such as tetrahydropyranyl), C ６～１０ aryl (e.g., phenyl) and 5-10 membered heteroaryl (e.g., 5-6 membered nitrogen-containing heteroaryl, 5-6 membered oxygen-containing heteroaryl, 5-6 membered sulfur-containing heteroaryl), optionally wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C １～６ Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), C １～６ Alkoxy (e.g., methoxy, ethoxy) and -C １～６ Alkylene-NH ２ (For example, -CH ２ -NH ２ ) independently substituted with one or more groups selected from the group consisting of: ２ is a covalent bond, 3- to 12-membered heterocyclyl (e.g., 3- to 6-membered saturated or unsaturated nitrogen-containing monoheterocyclyl, such as piperidinyl; 3- to 6-membered saturated or unsaturated oxygen-containing monoheterocyclyl, such as tetrahydropyranyl), and C ６～１０ aryl (e.g., phenyl), and optionally the heterocyclyl and aryl are selected from the group consisting of hydrogen, halogen, cyano, C １～６ Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), C １～６ Alkoxy (e.g., methoxy, ethoxy) and -C １～６ Alkylene-NH ２ (For example, -CH ２ -NH ２ each independently substituted with one or more groups independently selected from the group consisting of: ２ is selected from the group consisting of a covalent bond, piperidinyl, and tetrahydropyranyl; L ３ is a covalent bond or -C(O)-; L ４ is hydrogen, C １～６ Alkyl, -(O-CH ２ CH ２ ) ｎ -O-C １～６ Alkyl (e.g., -(O-CH ２ CH ２ ) ｎ -O-methyl, -((O-CH ２ CH ２ ) ｎ —O-ethyl) and —C １～６ Alkylene-(O-CH ２ CH ２ ) ｎ -O-C １～６ Alkyl (e.g., —CH ２ -(O-CH ２ CH ２ ) ｎ -O-methyl, -CH ２ -CH ２ -(O-CH ２ CH ２ ) ｎ -O-methyl, -CH ２ -(O-CH ２ CH ２ ) ｎ -O-ethyl, -CH ２ -CH ２ -(O-CH ２ CH ２ ) ｎ Optionally, each alkyl (e.g., methyl or ethyl) is selected from the group consisting of hydrogen, halogen, hydroxyl, —NH ２ , -NH-C(O)-OC １～６ Alkyl (e.g., —NH—C(O)—O-methyl), C １～６ Alkoxy (e.g., methoxy, ethoxy), carboxyl and -C(O)-O-C １～６ each n is independently selected from the group consisting of 1, 2, 3, 4, 5, 6, 7; preferably, L ４ is hydrogen, C １～６ Alkyl and -C １～６ Alkylene-(O-CH ２ CH ２ ) ｎ -O-C １～６ alkyl, optionally each alkyl being selected from the group consisting of hydroxyl, -NH ２ , C １～６ Alkoxy, carboxyl and -C(O)-O-C １～６ alkyl, each n is selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7; preferably, L ４ is hydrogen, C １～４ Alkyl and -C １～４ Alkylene-(O-CH ２ CH ２ ) ｎ -O-C １～４ alkyl, optionally each alkyl being selected from the group consisting of hydroxyl, -NH ２ , C １～４ Alkoxy, carboxyl and -C(O)-O-C １～４ and n is selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7.

3. A compound according to claim 1 or 2, or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof.

4. R ３ is hydrogen, or R ３ Is, -L ５ -L ６ -L ７ -L ８ Formula (In the formula, L ５ represents a covalent bond and, for example, -CH ２ --, --CH ２ -CH ２ --, --CH ２ -CH ２ -CH ２ -or-CH ２ -CH ２ -CH ２ -CH ２ - and other C １～６ alkylene; preferably, L ５ is a covalent bond; L ６ is a covalent bond, C ３～１０ Cycloalkyl (e.g., C ３～６ Cycloalkyl or C ５～６ cycloalkyl), 3-12 membered heterocyclyl (e.g., 3-6 membered saturated or unsaturated nitrogen-containing monoheterocyclyl (e.g., piperidinyl, tetrahydropyridyl (e.g., 1,2,3,6-tetrahydropyridyl), piperazinyl) or 3-6 membered saturated or unsaturated oxygen-containing monoheterocyclyl (e.g., tetrahydropyranyl)), C ６～１０ aryl (e.g., phenyl) and 5-10 membered heteroaryl (e.g., 5-6 membered nitrogen-containing heteroaryl, 5-6 membered oxygen-containing heteroaryl, 5-6 membered sulfur-containing heteroaryl), optionally wherein cycloalkyl, heterocyclyl, aryl and heteroaryl are selected from the group consisting of hydrogen, halogen, cyano, C １～６ Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), C １～６ Alkoxy (e.g., methoxy, ethoxy) and -C １～６ Alkylene-NH ２ (For example, -CH ２ -NH ２ each independently substituted with one or more groups selected from the group consisting of: ６ is selected from the group consisting of a covalent bond and 3-12 membered heterocyclyl (e.g., 3-6 membered saturated or unsaturated nitrogen-containing monoheterocyclyl (e.g., piperidinyl, tetrahydropyridyl (e.g., 1,2,3,6-tetrahydropyridyl), piperazinyl) or 3-6 membered saturated or unsaturated oxygen-containing monoheterocyclyl (e.g., tetrahydropyranyl)), where heterocyclyl is selected from the group consisting of hydrogen, halogen, cyano, C １～６ Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl), C １～６ Alkoxy (e.g., methoxy, ethoxy) and -C １～６ Alkylene-NH ２ (For example, -CH ２ -NH ２ Optionally substituted with one or more groups independently selected from the group consisting of: ６ is selected from the group consisting of a covalent bond, 1,2,3,6-tetrahydropyridyl, piperazinyl, and tetrahydropyranyl; L ７ is a covalent bond or -C(O)-, preferably L ７ is a covalent bond; L ８ is hydrogen, C １～６ Alkyl, -(O-CH ２ CH ２ ) ｑ -O-C １～６ Alkyl (e.g., -(O-CH ２ CH ２ ) ｑ -O-methyl, -(O-CH ２ CH ２ ) ｑ —O-ethyl) and —C １～６ Alkylene-(O-CH ２ CH ２ ) ｑ -O-C １～６ Alkyl (e.g., —CH ２ -(O-CH ２ CH ２ ) ｑ -O-methyl, -CH ２ -CH ２ -(O-CH ２ CH ２ ) ｑ -O-methyl, -CH ２ -(O-CH ２ CH ２ ) ｑ -O-ethyl, -CH ２ -CH ２ -(O-CH ２ CH ２ ) ｑ Optionally, each alkyl (e.g., methyl or ethyl) is selected from the group consisting of hydrogen, halogen, hydroxyl, —NH ２ , -NH-C(O)-OC １～６ Alkyl (e.g., —NH—C(O)—O-methyl), C １～６ Alkoxy (e.g., methoxy, ethoxy), carboxyl and -C(O)-O-C １～６ each q is independently selected from the group consisting of 1, 2, 3, 4, 5, 6, 7; preferably, L ８ is hydrogen, C １～６ Alkyl and -C １～６ Alkylene-(O-CH ２ CH ２ ) ｑ -O-C １～６ alkyl, optionally each alkyl being selected from the group consisting of hydroxyl, -NH ２ , C １～６ Alkoxy, carboxyl and -C(O)-O-C １～６ alkyl, and q is selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7; preferably, L ８ is hydrogen, C １～４ Alkyl and -C １～４ Alkylene-(O-CH ２ CH ２ ) ｑ -O-C １～４ alkyl, optionally each alkyl being selected from the group consisting of one or more C １～４ alkoxy groups, and q is selected from the group consisting of 1, 2, 3, 4, 5, 6, and 7; 4. The compound according to any one of claims 1 to 3, or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

5. Formula (II-A), Formula (II-B), Formula (II-C), Formula (II-D), Formula (II-E) or Formula (II-F): 【Chemistry 2】 (In the formula, R １ , R ２ and R ３ or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

6. Formula (III-A) or Formula (III-B): 【Chemistry 3】 (In the formula, R １ , R ３ , L ３ and L ４ wherein each of the groups is as defined in any one of claims 1 to 4), or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

7. Formula (IV-A) or Formula (IV-B): 【Chemistry 4】 (In the formula, R １ , L ３ , L ４ , L ７ and L ８ or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

8. Formula (VA) or Formula (V-B): 【Chemistry 5】 (In the formula, L ３ , L ４ , L ７ and L ８ or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

9. The following formula: 【Chemistry 6】 【change】 【change】 【change】 【change】 【change】 2. The compound of claim 1 having the structure: or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

10. The following formula: D-L' where D is a fragment of a compound according to any one of claims 1 to 9; L' is a linker; Preferably, L' is -L ９ -L １０ -L １１ (In the formula, L ９ is non-existent, or 【Chemistry 7】 Selected from: L １０ is absent or -(CH ２ CH ２ O) ｓ CH ２ CH ２ NH-, -(OCH ２ CH ２ ) ｓ NH- and -C(O)-(CH ２ CH ２ O) ｓ CH ２ CH ２ NH-; s is selected from the group consisting of integers from 1 to 10; L １１ teeth, 【Chemistry 8】 and Z １ is a chemical bond and 【Chemistry 9】 selected from the group consisting of: Z ２ is a chemical bond and C １～２０ alkylene; Z ３ is a chemical bond, C ２～６ Alkenylene and C ２～６ alkynylene; A is, 【Chemistry 10】 selected from the group consisting of: LG is a leaving group for nucleophilic substitution reactions, and is a halogen, nitro, benzenesulfonate, p-toluenesulfonate, trifluoromethanesulfonate, -S(O) ２ -R ７ and -S(O)-R ７ selected from the group consisting of: R ７ is C １～６ Alkyl and C １～６ haloalkyl); Preferably, L' is 【Chemistry 11】 【change】 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof.

11. The drug-linker has the following structure: 【Chemistry 12】 【change】 【change】 【change】 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, according to claim 10, selected from the group consisting of:

12. Formula (ISAC-I): 【Chemistry 13】 (In the formula, D is a fragment of a compound according to any one of claims 1 to 9; L is a fragment of the linker L', L' being as defined in claim 10; An Ab is an antibody capable of targeting a target antigen, optionally modified (e.g., modified with a cross-linking agent); z is selected from the group consisting of 1 to 10; Preferably, the immunostimulatory antibody conjugate has a DAR value, for example, of 1 to 10, for example: 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 2 to 3, 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 2 to 10, 3 to 4, 3 to 5, 3 to 6, 3 to 7, 3 to 8, 3 to 9, 3 to 10, 4 to 5, 4 to 6, 4 to 7, 4 to 8, 4 to 9, 4 to 10, 5 to 6, 5 to 7, 5 to 8, 5 to 9, 5 to 10, 6 to 7, 6 to 8, 6 to 9, 6 to 10, 7 to 8, 7 to 9, 7 to 10, 8 to 9, 8 to 10, or 9 to 10, preferably 1 to 8, for example 1.

0. Up to 1.5, 1.0 to 2.0, 1.0 to 2.5, 1.0 to 3.0, 1.0 to 3.5, 1.0 to 4.0, 1.0 to 4.5, 1.0 to 5.0, 1.0 to 5.5, 1.0 to 6.0, 1.0 to 6.5, 1.0 to 7.0, 1.0 to 7.5, 1.0 to 8.0, 1.5 to 2.0, 1.5 to 2.5, 1.5-3.0, 1.5-3.5, 1.5-4.0, 1.5-4.5, 1.5-5.0, 1.5-5.5, 1.5-6.0, 1.5-6.5, 1.5-7.0, 1.5-7.5, 1.5-8.0, 2.0-2.5, 2.0-3.0, 2.0-3.5, 2.0-4.0, 2.0-4 .5, 2.0-5.0, 2.0-5.5, 2.0-6.0, 2.0-6.5, 2.0-7.0, 2.0-7.5, 2.0-8.0, 2.5-3.0, 2.5-3.5, 2.5-4.0, 2.5-4.5, 2.5-5.0, 2.5-5.5, 2.5-6.0, 2.5-6.5, 2. .5 to 7.0, 2.5 to 7.5, 2.5 to 8.0, 3.0 to 3.5, 3.0 to 4.0, 3.0 to 4.5, 3.0 to 5.0, 3.0 to 5.5, 3.0 to 6.0, 3.0 to 6.5, 3.0 to 7.0, 3.0 to 7.5, 3.0 to 8.0, 3.5 to 4.0, 3.5 to 4.5, 3.5 to 5. 0, 3.5-5.5, 3.5-6.0, 3.5-6.5, 3.5-7.0, 3.5-7.5, 3.5-8.0, 4.0-4.5, 4.0-5.0, 4.0-5.5, 4.0-6.0, 4.0-6.5, 4.0-7.0, 4.0-7.5, 4.0-8.0, 4.5-5.0, 4.5 Up to 5.5, 4.5 to 6.0, 4.5 to 6.5, 4.5 to 7.0, 4.5 to 7.5, 4.5 to 8.0, 5.0 to 5.5, 5.0 to 6.0, 5.0 to 6.5, 5.0 to 7.0, 5.0 to 7.5, 5.0 to 8.0, 5.5 to 6.0, 5.5 to 6.5, 6.0 to 7.0, 6.0 to 7.5,6.0-8.0, 6.5-7.0, 6.5-7.5, 6.5-8.0, 7.0-7.5, 7.0-8.0 or 7.5-8.0), or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof.

13. Use of a compound in the manufacture of an immunostimulatory antibody conjugate, said compound being selected from the compounds according to any one of claims 1 to 9.

14. A method for preparing an immunostimulatory antibody conjugate comprising the step of using a compound according to any one of claims 1 to 9.

15. A pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound according to any one of claims 1 to 9 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, a drug-linker according to claim 10 or 11 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, or an immunostimulatory antibody conjugate according to claim 12, and one or more pharma- ceutically acceptable carriers.

16. Use of a compound according to any one of claims 1 to 9 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, a drug-linker according to claim 10 or 11 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, an immunostimulatory antibody conjugate according to claim 12, or a pharmaceutical composition according to claim 15, in the manufacture of a medicament for the prophylaxis and / or treatment of a disease mediated by TLR7 and / or TLR8 (e.g. a tumor).

17. 16. A method for preventing and / or treating a disease mediated by TLR7 and / or TLR8 (e.g. tumors) comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 9 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, a drug-linker according to claim 10 or 11 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, an immunostimulatory antibody conjugate according to claim 12, or a pharmaceutical composition according to claim 15.

18. A kit comprising a compound according to any one of claims 1 to 9 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, a drug-linker according to claim 10 or 11 or a pharma- ceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, an immunostimulatory antibody conjugate according to claim 12, or a pharmaceutical composition according to claim 15.

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