Pyridine [3, 2-d] pyrimidine-2-amine derivative as well as preparation method and application thereof

Abstract

The invention discloses a pyridine [3, 2-d] pyrimidine-2-amine derivative as well as a preparation method and application thereof. The pyridine [3, 2-d] pyrimidine-2-amine derivative can be used as a TLR7 / 8 dual agonist and is used for treating diseases related to TLR activity, such as diseases, tumors, inflammatory diseases and the like caused by virus (such as HBV (Hepatitis B Virus) and HIV (Human Immunodeficiency Virus)) infection. Particularly, the pyridine [3, 2-d] pyrimidine-2-amine derivative provided by the invention has better physicochemical properties, druggability, heart safety and oral bioavailability, and has very good research and application values in the field of medicines.

Description

Technical Field

[0001] The present invention relates to the technical field of Toll-like receptor agonists (TLRs), and in particular to a pyridine [3,2-d] pyrimidine-2-amine derivative or a salt thereof used as a TLR7 / 8 dual agonist, a preparation method thereof, a pharmaceutical composition, and an application thereof in preparing a drug for treating diseases, tumors, and the like caused by viral (e.g., HBV, HIV) infection. Background Art

[0002] Toll-like receptors (TLR1, 2, 3-13) are a very important class of pattern recognition receptors that specifically recognize a variety of pathogen-associated molecular patterns (PAMPs) in the body. Most of them are on the cell surface, including TLR1, TLR2, TLR4, TLR5, TLR6, and TLR10, while some are expressed in endosomes, including TLR3, TLR7, TLR8, and TLR9 (Semin Immunol. 2007, 19(1): 24-32.).

[0003] TLR7 and TLR8 can specifically recognize single-stranded RNA of pathogens. Therefore, exogenous or endogenous ligands can activate TLR7 and 8 after binding to TLR7 / 8, causing a series of signal cascade reactions mediated by MyD88, activating the innate immune response. TLR7 / 8 is mainly expressed on innate immune cells, such as dendritic cells, macrophages and NK cells. Activating TLT7 / 8 on these cells can release proinflammatory factors and chemokines, thereby activating acquired immunity and killing or clearing cells infected with viruses. Therefore, TLR7 and 8 are also important bridges connecting natural immunity and acquired immunity. In particular, activation of TLR7 on plasma dendritic cells leads to the secretion of type I interferons (IFN-α / β) and activation of TLR8 on myeloid dendritic cells leads to the production of type II interferons (IFN-γ), TNF-α and IL-12, Th1-polarized cytokines that can induce T helper-1 type adaptive immune responses (J. Med. Chem. 2021, 64, 7507-7532).

[0004] Activation of TLR7 can activate antigen-presenting cells such as dendritic cells and macrophages, present tumor cell-related antigens to cytotoxic T lymphocytes, indirectly activate T cell immunity, and kill tumor cells. Activation of TLR8 can activate natural killer cells and play an anti-tumor activity. More importantly, activation of TLR8 can reprogram regulatory T cells (Treg cells) through the glycolytic pathway and reverse their immunosuppressive activity (Cell Metab. 2019, 29 (1): 103-123.e5.). In addition, activation of TLR8 can induce apoptosis of immunosuppressive myeloid cells (MDSC), thereby enhancing immune-mediated anti-tumor activity (J Leukoc Biol. 2018, 103 (1): 157-164.). In summary, TLR7 / 8 agonists have the effect of tumor immunotherapy.

[0005] On the other hand, TLR7 selective agonists (such as GS-9620) have shown significant therapeutic effects in the treatment of hepatitis B and AIDS in clinical trials. At the same time, TLR8 agonists also have significant effects in the treatment of viral infections (such as hepatitis B). In HIV infection, latent viral reservoirs, especially latently infected CD4 + The viral reservoir in T lymphocytes is the main obstacle in the treatment of AIDS. It can cause viral rebound, thus hindering the effectiveness of antiviral therapy (ART). Therefore, AIDS patients need to take medication for life to suppress the virus and alleviate other manifestations. At present, cocktail therapy is widely used in the clinic to treat AIDS patients. At present, immunotherapy strategies are a potential means to cure AIDS. One of the strategies is that activating viral reservoirs may make them more susceptible to immune-mediated killing. According to mechanism studies reported in relevant literature, the cytokine INF-α released by TLR7 activation and TNF-α released by TLR8 activation can both activate latent viruses in AIDS-infected cells. Therefore, simultaneous activation of TLR7 and TLR8 may have a synergistic effect in activating the latent AIDS virus reservoir (Front Microbiol. 2023, 14: 1033448). Summary of the invention

[0006] The present invention provides a pyridine [3,2-d] pyrimidine-2-amine derivative or a salt thereof, which can be used as a TLR7 / 8 receptor and has the characteristics of high selectivity, strong activity and good safety. The present invention also provides a preparation method of the pyridine [3,2-d] pyrimidine-2-amine derivative or a salt thereof, a pharmaceutical composition and an application thereof.

[0007] In a first aspect of the present invention, a pyridinium [3,2-d] pyrimidin-2-amine derivative is provided, which has the following structure:

[0008]

[0009]

[0010] in,

[0011] L1 is C1-C6 alkylene or C3-C6 cycloalkylene;

[0012] L2 is C1-C6 alkylene or -C(O)-;

[0013] Ring A is an aromatic ring or a heterocyclic ring, for example:

[0014] R0 is selected from: H, C1-C6 alkyl, C1-C6 haloalkyl, -O(C0-C6 alkyl), halogen, cyano, -N(C0-C6 alkyl)(C0-C6 alkyl), heterocyclyl;

[0015] R1 is selected from: H, -O(C0-C6 alkyl), -(C1-C6 alkylene)-OH, -OC(O)(C0-C6 alkyl), and a basic tertiary amine group;

[0016] R2 and R3 are independently selected from: H, C1-C6 alkyl, C1-C6 haloalkyl, -(C1-C6 alkylene)-COO-(C0-C6 alkyl), 4-7 membered heterocyclyl (especially 5- or 6-membered saturated heterocyclyl, such as nitrogen-containing heterocyclyl or oxygen-containing heterocyclyl); or R2 and R3 together with the nitrogen atom to which they are commonly attached form a heterocycle, which is optionally substituted by one or more groups selected from the following: halogen, C1-C6 alkyl, C1-C6 haloalkyl, -(C0-C6 alkylene)-OH, -(C0-C6 alkylene)- wherein the heterocyclyl is optionally substituted by one or more groups selected from the group consisting of halogen, C1-C6 alkyl, C1-C6 haloalkyl, -(C0-C6 alkylene)-OH, -(C0-C6 alkylene)-COO-(C0-C6 alkyl), -(C0-C6 alkylene)-N(C0-C6 alkyl)(C0-C6 alkyl), -(C0-C6 alkylene)-heterocyclyl; wherein the heterocyclyl is optionally substituted by one or more groups selected from the group consisting of halogen, C1-C6 alkyl, C1-C6 haloalkyl, -(C0-C6 alkylene)-OH, -(C0-C6 alkylene)-COO-(C0-C6 alkyl), -(C0-C6 alkylene)-N(C0-C6 alkyl)(C0-C6 alkyl).

[0017] Specifically, R0 can be selected from: H, methyl, ethyl, methoxy, halogen, cyano, hydroxyl, amine, trifluoromethyl, difluoromethyl,

[0018] In some embodiments of the present invention, R0 is H.

[0019] In some embodiments of the present invention, Some selected from:

[0020] in particular

[0021] In some embodiments of the present invention, ring A is a benzene ring, and the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0022]

[0023] In some embodiments of the present invention, L1 is selected from:

[0024] In some embodiments of the present invention, L2 is selected from:

[0025] In some embodiments of the invention, R1 is H or -O(C1-C3 alkyl), particularly H or methoxy.

[0026] In some embodiments of the present invention, R2 is selected from: H, C1-C3 alkyl (eg, methyl), C1-C3 haloalkyl (eg, -CF3, -CH2CF3).

[0027] In some embodiments of the present invention, R3 is selected from: C1-C3 alkyl (e.g., methyl), -(C1-C3 alkylene)-COO-(C0-C3 alkyl) (e.g., ), 5- or 6-membered saturated heterocyclic group (e.g. ).

[0028] In some embodiments of the present invention, R2 is C1-C3 alkyl (eg, methyl), and R3 is C1-C3 alkyl (eg, methyl).

[0029] In some embodiments of the present invention, R2 is selected from: H, C1-C3 alkyl (e.g., methyl), C1-C3 haloalkyl (e.g., -CF3, -CH2CF3), R3 is -(C1-C3 alkylene)-COO-(C0-C3 alkyl) (e.g., ).

[0030] In some embodiments of the present invention, R2 is H, R3 is a 4-6 membered saturated heterocyclic group (eg ).

[0031] In some embodiments of the present invention, R2 and R3 together with the nitrogen atom to which they are attached form a 4-10 membered saturated heterocyclic ring (eg

[0032] ), the heterocycle is optionally substituted by one or more groups selected from the group consisting of halogen (e.g., F), C1-C3 alkyl (e.g., methyl), C1-C3 haloalkyl (e.g., trifluoromethyl), -(C0-C3 alkylene)-OH (e.g., -OH, ), -(C0-C3 alkylene)-COO-(C0-C3 alkyl) (e.g. -COOH,

[0033] ), -(C0-C6 alkylene)-N(C0-C6 alkyl)(C0-C6 alkyl)(e.g. -NH2, ), -(C0-C6 alkylene)-(4-10 membered saturated heterocyclic group), wherein the 4-10 membered saturated heterocyclic group (e.g. ) is optionally substituted by one or more groups selected from the following: halogen, C1-C3 alkyl, C1-C3 haloalkyl, -(C0-C3 alkylene)-OH, -(C0-C3 alkylene)-COO-(C0-C3 alkyl), -(C0-C3 alkylene)-N(C0-C3 alkyl)(C0-C3 alkyl).

[0034] In some embodiments of the present invention, The section has the following structure:

[0035] In some embodiments of the present invention, the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0036]

[0037] in,

[0038] Ring B is a 4-10 membered saturated heterocyclic ring (including monocyclic and polycyclic rings, such as fused bicyclic rings and spirocyclic rings);

[0039] R4 is one or more independent substituents on the B ring, which are independently selected from: H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -(C0-C6 alkylene)-OH, -(C0-C6 alkylene)-COO-(C0-C6 alkyl), -(C0-C6 alkylene)-N(C0-C6 alkyl)(C0-C6 alkyl), -(C0-C6 alkylene)-(4-10 membered saturated heterocyclyl), wherein the 4-10 membered saturated heterocyclyl is optionally substituted by one or more groups selected from the following: halogen, C1-C3 alkyl, C1-C3 haloalkyl, -(C0-C3 alkylene)-OH, -(C0-C3 alkylene)-COO-(C0-C3 alkyl), -(C0-C3 alkylene)-N(C0-C3 alkyl)(C0-C3 alkyl).

[0040] In some embodiments of the invention, Ring B is selected from:

[0041] In some embodiments of the present invention, the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0042]

[0043] in,

[0044] X is selected from: single bond, N, O, CH, C(O), S, S(O), S(O)2;

[0045] R4 is one or more independent substituents on the ring, which are independently selected from: H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -(C0-C6 alkylene)-OH, -(C0-C6 alkylene)-COO-(C0-C6 alkyl), -(C0-C6 alkylene)-N(C0-C6 alkyl)(C0-C6 alkyl), -(C0-C6 alkylene)-(4-10 membered saturated heterocyclyl), wherein the 4-10 membered saturated heterocyclyl is optionally substituted with one or more groups selected from the following: halogen, C1-C3 alkyl, C1-C3 haloalkyl, -(C0-C3 alkylene)-OH, -(C0-C3 alkylene)-COO-(C0-C3 alkyl), -(C0-C3 alkylene)-N(C0-C3 alkyl)(C0-C3 alkyl).

[0046] In some embodiments of the present invention, the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0047]

[0048] Wherein, R4 is selected from: H, C1-C6 alkyl, -(C1-C6 alkylene)-OH, -(C1-C6 alkylene)-COO-(C0-C6 alkyl).

[0049] In some embodiments of the present invention, the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0050]

[0051] In some embodiments of the present invention, in formula IV-1 and IV-2, R4 is H.

[0052] In some embodiments of the present invention, in formula IV-1 and IV-2, R4 is a C1-C6 alkyl group, such as methyl, ethyl, n-propyl, isopropyl.

[0053] In some embodiments of the present invention, in formula IV-1 and IV-2, R4 is -(C1-C6 alkylene)-OH, such as

[0054] In some embodiments of the present invention, in formula IV-1 and IV-2, R1 is H. In other embodiments of the present invention, in formula IV-1 and IV-2, R1 is C1-C3 alkoxy, such as methoxy.

[0055] In some embodiments of the present invention, in formula IV-1 and IV-2, L1 is selected from: in particular

[0056] In some embodiments of the present invention, in formula IV-1 and IV-2, L2 is C1-C3 alkylene, especially methylene. In some embodiments of the present invention, in formula IV-1 and IV-2, L2 is

[0057] In some embodiments of the present invention, the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0058]

[0059] Wherein, R4 is one or more independent substituents on the ring, which are independently selected from: H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -(C0-C6 alkylene)-OH, -(C0-C6 alkylene)-COO-(C0-C6 alkyl), -(C0-C6 alkylene)-N(C0-C6 alkyl)(C0-C6 alkyl), -(C0-C6 alkylene)-(4-10 membered saturated heterocyclic group), wherein the The 4-10 membered saturated heterocyclic group is optionally substituted by one or more groups selected from the following: halogen, C1-C3 alkyl, C1-C3 haloalkyl, -(C0-C3 alkylene)-OH, -(C0-C3 alkylene)-COO-(C0-C3 alkyl), -(C0-C3 alkylene)-N(C0-C3 alkyl)(C0-C3 alkyl), in particular -(C1-C3 alkylene)-COO-(C0-C3 alkyl).

[0060] Specifically, in formula V, each R4 is independently selected from: H, halogen (such as F), C1-C6 alkyl, -(C0-C6 alkylene)-OH, -(C0-C6 alkylene)-COO-(C0-C6 alkyl); or two R4 connected to the same carbon atom together with the carbon atom form a C3-C6 carbocycle (such as a C3 saturated carbocycle).

[0061] In some embodiments of the present invention, the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0062]

[0063] In some embodiments of the present invention, in formula V and V-1, R4 and R4' are H.

[0064] In some embodiments of the present invention, in formula V and V-1, R4' is H, R4 is -(C0-C6 alkylene)-COO-(C0-C6 alkyl), such as

[0065] In some embodiments of the present invention, in formula V and V-1, R4 and R4' are independently selected from: halogen (such as F), C1-C6 alkyl (such as methyl).

[0066] In some embodiments of the present invention, in formula V and V-1, R1 is H. In other embodiments of the present invention, in formula V and V-1, R1 is C1-C3 alkoxy, such as methoxy.

[0067] In some embodiments of the present invention, in Formula V, V-1, L1 is selected from: in particular

[0068] In some embodiments of the present invention, in formula V and V-1, L2 is C1-C3 alkylene, especially methylene. In some embodiments of the present invention, in formula V and V-1, L2 is

[0069] In some embodiments of the present invention, the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0070]

[0071] In some embodiments of the present invention, the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0072]

[0073] In some embodiments of the present invention, in formula VI and VI-1, R2 is a C1-C3 alkyl group (eg, methyl group), and R3 is a C1-C3 alkyl group (eg, methyl group).

[0074] In some embodiments of the present invention, in formula VI, VI-1, R2 is H, and R3 is selected from:

[0075] In some embodiments of the present invention, in formula VI and VI-1, R2 is C1-C3 haloalkyl, such as -CF3, -CH2CF3, and R3 is -(C1-C3 alkylene)-COO-(C0-C3 alkyl) (e.g. ).

[0076] In some embodiments of the present invention, in formula VI, VI-1, R2 is H, R3 is -(C1-C3 alkylene)-COO-(C0-C3 alkyl), such as

[0077] In some embodiments of the present invention, in formula VI, VI-1, L1 is selected from: in particular

[0078] In some embodiments of the present invention, in formula VI and VI-1, R1 is H. In other embodiments of the present invention, in formula VI and VI-1, R1 is C1-C3 alkoxy, such as methoxy.

[0079] In some embodiments of the present invention, the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0080]

[0081] In some embodiments of the present invention, the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0082]

[0083] In some embodiments of the present invention, in formula VII and VII-1, R2 is a C1-C3 alkyl group (eg, methyl group), and R3 is a C1-C3 alkyl group (eg, methyl group).

[0084] In some embodiments of the present invention, in formula VII, VII-1, R2 is H, and R3 is selected from:

[0085] In some embodiments of the present invention, in formula VII and VII-1, R2 is C1-C3 haloalkyl, such as -CF3, -CH2CF3, and R3 is -(C1-C3 alkylene)-COO-(C0-C3 alkyl) (e.g. ).

[0086] In some embodiments of the present invention, in formula VII, VII-1, R2 is H, R3 is -(C1-C3 alkylene)-COO-(C0-C3 alkyl), such as

[0087] In some embodiments of the present invention, in Formula VII, VII-1, L1 is selected from: in particular

[0088] In some embodiments of the present invention, in formula VII and VII-1, R1 is H. In other embodiments of the present invention, in formula VII and VII-1, R1 is C1-C3 alkoxy, such as methoxy.

[0089] In some embodiments of the present invention, the pyridin[3,2-d]pyrimidin-2-amine derivative has the following structure:

[0090]

[0091]

[0092]

[0093] In the second aspect of the present invention, provided are pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, solvates, and isotopic derivatives of the pyridinium [3, 2-d] pyrimidin-2-amine derivatives of the first aspect.

[0094] In some embodiments of the invention, the stereoisomer has the following structure:

[0095]

[0096]

[0097] In the third aspect of the present invention, there is provided a method for preparing the pyridine [3,2-d] pyrimidine-2-amine derivative according to the first aspect, comprising the following steps:

[0098] Compound 1 and 2 react to obtain the target compound

[0099]

[0100] Among them, R L1 For the leaving group.

[0101] In some embodiments of the present invention, L2 is C1-C3 alkylene, especially methylene, R L1 The method comprises step (I): mixing compound 2, solvent I, compound 1 and catalyst I, reacting, separating and purifying.

[0102] Specifically, the solvent I is selected from: acetonitrile, ethanol, methanol, dimethylformamide, etc., one or more combinations thereof, especially acetonitrile.

[0103] Specifically, catalyst I is selected from: a combination of one or more of alkali, copper salt, silver salt, etc.; specifically, the alkali can be sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, sodium iodide, etc.; in some embodiments of the present invention, catalyst I includes potassium carbonate and sodium iodide.

[0104] Specifically, the reaction temperature is 50-100°C, for example, 50, 55, 60, 65, 70, 80, 90, 100°C, especially 55-65°C.

[0105] Specifically, the reaction time is 1-6 hours, for example, 1, 1.5, 2, 2.5, 3, 4, 5, 6 hours.

[0106] Specifically, the above reaction can be carried out under stirring.

[0107] In some embodiments of the present invention, L2 is -C(O)-, R L1 OH or OM + , M is an alkali metal (such as lithium, sodium, potassium), and the method comprises step (II): mixing compound 2, solvent II, compound 1, and condensation reagent II, reacting, and separating and purifying.

[0108] Specifically, solvent II is selected from: DMF, dichloromethane, DMSO, NMP, especially DMF.

[0109] Specifically, the condensation reagent II is selected from: HATU, TBTU, TPTU, PyBOP, EDC, DIC, etc., especially HATU.

[0110] Furthermore, the above reaction system may also include an acylation catalyst or activator II, such as DIEA, DMAP, HOBt, etc.

[0111] Specifically, the reaction temperature is 0-50°C, for example 0, 5, 10, 15, 20, 25, 30, 35, 40, 50°C, especially room temperature.

[0112] Specifically, the reaction time is 1-6 hours, for example, 1, 1.5, 2, 2.5, 3, 4, 5, 6 hours.

[0113] Specifically, the above reaction can be carried out under stirring.

[0114] Specifically, the method may further include the following reaction steps:

[0115] Compound 4, solvent III, compound 3, and catalyst III are mixed, reacted, separated, and purified.

[0116]

[0117] Among them, R L2 , R L3 is a leaving group (such as halogen), R L1 ' is an ester group (eg, a methyl ester group).

[0118] In some embodiments of the present invention, R L2 Cl, R L3 For Br.

[0119] Specifically, the solvent III is selected from: tetrahydrofuran, diethyl ether, dioxane, xylene, especially tetrahydrofuran.

[0120] Specifically, catalyst III is a metal catalyst. In some embodiments of the present invention, catalyst III includes a nickel chloride reagent, such as bis(triphenylphosphine)nickel chloride; more specifically, catalyst III also includes other metal catalysts, such as zinc powder.

[0121] Specifically, the reaction temperature is 0-50°C, for example 0, 5, 10, 15, 20, 25, 30, 35, 40, 50°C, especially room temperature.

[0122] Specifically, the reaction time is 6-30 hours, for example 6, 12, 18, 24, 30 hours.

[0123] Specifically, the above reaction is carried out under an inert atmosphere (eg, nitrogen).

[0124] Specifically, the above reaction can be carried out under stirring.

[0125] Specifically, the method may further include the following reaction steps:

[0126] Compound 5, solvent IV, 2-methoxyethylamine and catalyst IV are mixed, reacted, separated and purified.

[0127]

[0128] Specifically, solvent IV is selected from: DMF, dichloromethane, DMSO, NMP, especially DMF.

[0129] Specifically, the catalyst IV is selected from: HATU, TBTU, TPTU, PyBOP, EDC, DIC, etc., especially HATU.

[0130] Furthermore, the above reaction system may also include an activator IV, such as DIEA, DMAP, HOBt, etc.

[0131] Specifically, the reaction temperature is 0-50°C, for example 0, 5, 10, 15, 20, 25, 30, 35, 40, 50°C, especially room temperature.

[0132] Specifically, the reaction time is 6-30 hours, for example 6, 12, 18, 24, 30 hours.

[0133] Specifically, the above reaction can be carried out under stirring.

[0134] Specifically, the method may further include the following reaction steps:

[0135] Compound 6, solvent V, guanidine hydrochloride, catalyst V and a base are mixed, reacted, separated and purified.

[0136]

[0137] Among them, R L4 is a leaving group (eg halogen).

[0138] Specifically, solvent V is selected from: DMF, DMSO, THF, acetonitrile, especially DMF.

[0139] Specifically, the catalyst V is selected from: copper, copper iodide, cuprous iodide, cuprous oxide, palladium chloride or palladium acetate, especially cuprous oxide.

[0140] In particular, the base is selected from: sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, especially cesium carbonate.

[0141] Specifically, the reaction temperature is 100-150°C, for example, 100, 110, 120, 130, 140, 150°C.

[0142] Specifically, the reaction time is 6-30 hours, for example 6, 12, 18, 24, 30 hours.

[0143] Specifically, the above reaction can be carried out under stirring.

[0144] In the fourth aspect of the present invention, a pharmaceutical composition is provided, comprising the pyridine [3,2-d] pyrimidine-2-amine derivative or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative thereof as described in the first aspect, and one or more pharmaceutically acceptable excipients.

[0145] Specifically, the pharmaceutically acceptable excipients include, but are not limited to, sweeteners, diluents, stabilizers, emulsifiers, dispersants, preservatives, colorants, flavor enhancers, surfactants, wetting agents, disintegrants, suspending agents, isotonic agents, and solvents.

[0146] Specifically, the pharmaceutical composition can be prepared into tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, solutions, suppositories, injections, inhalants, and gels.

[0147] Specifically, the routes of administration of the pharmaceutical composition include, but are not limited to, oral, rectal, transmucosal, topical, transdermal, inhalation, parenteral, sublingual, intravaginal, intranasal, intramuscular, subcutaneous, and intravenous administration.

[0148] In some embodiments of the present invention, the pharmaceutical composition is administered orally.

[0149] Specifically, the pharmaceutical composition further contains at least one other therapeutic agent, which is selected from chemotherapeutic agents, immunotherapeutic agents, anti-angiogenic agents, cytokines, hormones, polynucleotides, antibodies, and immunologically active fragments.

[0150] Specifically, the antibodies include, but are not limited to: PRO542, which is an anti-HIV gp120 antibody fused to CD4 (Progenics / Genzyme Transgenics); MDX-010 (Medarex, New Jersey), which is a humanized anti-CTLA-4 antibody; SYNAGIS (MedImmune, Maryland), which is a humanized anti-respiratory syncytial virus (RSV) monoclonal antibody; HERCEPTIN (trastuzumab) (Genentech, California), which is a humanized anti-HER2 monoclonal antibody; Humanized anti-CD18 F(ab')2 (Genentech); CDP860, which is a humanized anti-CD18 F(ab')2 (Celltech, UK); Ostavir, which is a human anti-hepatitis B virus antibody (Protein Design Lab / Novartis); PROTOVIRTM, which is a humanized anti-CMV IgG1 antibody (Protein Design Lab / Novartis); MAK-195 (SEGARD), which is a murine anti-TNF-α F(ab')2 (Knoll Pharmaceuticals / BASF); llPharma / BASF); IC14, which is an anti-CD14 antibody (ICOS Pharmaceuticals (ICOS Pharm)); humanized anti-VEGF IgG1 antibody (Genentech); OVAREXM, which is a mouse anti-CA125 antibody (Altarex); PANOREXM, which is a mouse anti-17-IA cell surface antigen IgG2a antibody (GlaxoWellcome / Centocor); BEC2, which is a mouse anti-idiotype (GD3 epitope) IgG antibody (ImClone System); IMC-C2 25, which is a chimeric anti-EGFR IgG antibody (Immunoclonal Systems); VITAXIN™, which is a humanized anti-αVβ3 integrin antibody (Applied Molecular Evolution / MedImmune); Campath 1H / LDP-03, which is a humanized anti-CD52 IgG1 antibody (Leukosite); SmartM195, which is a humanized anti-CD33 IgG antibody (Protein Design Lab / Kanebo);RITUXAN™ (rituximab™), a chimeric anti-CD20 IgG1 antibody (IDEC Pharmaceuticals / Genentech, Roche / Zettyaku); LYMPHOCIDE™, a humanized anti-CD22 IgG antibody (Immunomedics); SmartID10, a humanized anti-HLA antibody (Protein Design Laboratories); ONCOLYM™ (Lym-1), a radiolabeled mouse anti-HLADR antibody (Techniclone); ABX-IL8, a human anti-IL8 antibody (Abgeni x)); anti-CD11a, a humanized IgG1 antibody (Genentech / Xoma); ICM3, a humanized anti-ICAM3 antibody (ICOS Pharmaceuticals); IDEC-114, a primatized anti-CD80 antibody (IDEC Pharmaceuticals / Mitsubishi); ZEVALIN™, a radiolabeled murine anti-CD20 antibody (IDEC / Schering AG); IDEC-131, a humanized anti-CD40L antibody (IDEC / Eisai); IDEC-151, a primatized anti-CD4 antibody (IDEC); IDEC-152, a primatized anti-CD4 antibody (IDEC); SMART anti-CD3, a humanized anti-CD3 IgG (Protein Design Laboratories); 5G1.1, a humanized anti-complement factor 5 (C5) antibody (Alexion Pharm); D2E7, a humanized anti-TNF-α antibody (CAT / BASF); CDP870, a humanized anti-TNF-α Fab fragment (Celtec); IDEC-151, a primatized anti-CD4 IgG1 antibody (IDEC Pharmaceuticals / SmithKline Beecham); MDX-CD4, a humanized anti- CD4 IgG antibody (Medarex / Eisai / Genmab); CDP571, a humanized anti-TNF-α IgG4 antibody (Celtech); LDP-02, a humanized anti-α4β7 antibody (Luxeter / Genentech); OrthoCloneOKT4A, a humanized anti-CD4 IgG antibody (OrthoBiotech); ANTOVATM, a humanized anti-CD40L IgG antibody (Biogen); ANTEGRENTM, a humanized anti-VLA-4 IgG antibody (Elan);MDX-33, a human anti-CD64 (FcγR) antibody (Medlex / Centeon); SCH55700, a humanized anti-IL-5 IgG4 antibody (Celtech / Schering); SB-240563 and SB-240683, humanized anti-IL-5 and anti-IL-4 antibodies, respectively (SmithKline Beecham); rhuMab-E25, a humanized anti-IgE IgG1 antibody (Genentech / Norvartis / Tanox Biosystems); ABX-CBL, a mouse anti-CD-147 IgM antibody (Abgenix); BTI-322, a rat anti-CD2 IgG antibody (Medical Immunology / BioTransplant); Odoclone / OKT3 (Orth Biosystems); oclone / OKT3), a mouse anti-CD3 IgG2a antibody (OrthoBiotech); SIMULECTTM, a chimeric anti-CD25 IgG1 antibody (Novartis); LDP-01, a humanized anti-β2-integrin IgG antibody (Luxet); anti-LFA-1, a mouse anti-CD18 F(ab′)2 (Pasteur-Merieux / Immunotech); CAT-152, a human anti-TGF-β2 antibody (Cambridge AbTech); CorsevinM, a chimeric anti-factor VII antibody (Santok); MDX-1106, a PD-1 antibody (Bristol-Myers Squibb); MDX-1105, a PDL1 antibody (Roche). ;

[0151] In some embodiments of the invention, the antibody is an HIV antibody.

[0152] In some embodiments of the present invention, the antibody is a PD-1 antibody.

[0153] Specifically, the anticancer agent includes but is not limited to: acivicin, aclarubicin, acodazole hydrochloride, aclonine, adolesin, aldesleukin, hexamethylmelamine, ambromycin, amenthranol acetate, aminoglutethimide, amsacrine, anastrozole, anthramycin, asparaginase, troline, azacitidine, azatepa, azomycin, batimastat, benzotepa, bicalutamide, bisantrene hydrochloride, binefad dimesylate, bisezole, bleomycin sulfate, buquina sodium, bropiridamine, busulfan, actinomycin C, caprotestosterone, carcetamide, carbetim, carboplatin, carmustine, carrubicin hydrochloride, carzelesin, cedifingol, chlorambucil, cyromycin, cisplatin, cladribine, clinatol mesylate, cyclophosphamide, cytarabine, Dacarbazine, actinomycin D, daunorubicin hydrochloride, decitabine, dextromaplatin, dezaguanine, dezaguanine mesylate, diazocone, docetaxel, doxorubicin, doxorubicin hydrochloride, droloxifene, droloxifene citrate, drolostanone propionate, dazomicin, edatrexa, eflornithine hydrochloride, elsamitrucin, enloplatin, enprobamate, epiripridine, epirubicin hydrochloride, erbulazole, esorubicin hydrochloride, estramustine, estramustine sodium phosphate, etaninidazole, etoposide, etoposide phosphate, chlorpheniramine, fadrozole hydrochloride, fazarabine, fenretinide, azauridine, fludarabine phosphate, fluorouracil, flucitabine, fosquatone, fostramecin sodium, gemcitabine, gemcitabine hydrochloride, hydroxyurea, idarubicin hydrochloride, ifosfamide , imofosine, interleukin II (including recombinant interleukin II or rIL2), interferon α2a, interferon α-2b, interferon α-n1, interferon α-n3, interferon β-Ia, interferon γ-Ib, iproplatin, irinotecan hydrochloride, lanreotide acetate, letrozole, leuprolide acetate, lirazo hydrochloride, lometrexol sodium, lomustine, losoxantrone hydrochloride, masoprofen, maytansine, nitrogen mustard hydrochloride, progesterone acetate, melengestrol acetate, phenylalanine mustard, menolipir, mercaptopurine, methotrexate, methotrexate sodium, chlorpheniramine, metodepa, mibutamide, mitokacin, mitoclopramide, mitoxantrone, mitoxetine, mitosan, mitoxetine, mitosep, mitotane, mitoxantrone hydrochloride, mycophenolic acid, nocodazole, nocodazole Lathromycin, Omaplatin, Oxysulen, Paclitaxel, Pegaspargase, Pelimycin, Namustine, Pelimycin Sulfate, Perfosfamide, Pipobroman, Piposufan, Pyroxantrone Hydrochloride, Plicamycin, Promectin, Porfil Sodium, Porfiromycin, Prednimustine, Procarbazine Hydrochloride, Puromycin, Puromycin Hydrochloride, Pyrazofuranin, Liboadenosine, Roglulimide, Safingol, Safingol Hydrochloride, Semustine, Simtraqin, Sodium Phosacetyl Aspartate, Sparamycin, Spirogermanium Hydrochloride, Spiromustine, Spiroplatin, Streptozocin, Streptozocin, Sulfonyl Chloride, Talithromycin, Tecogalan Sodium, Tegafur, Tiloxantrone Hydrochloride, Temoporfin, Teniposide, Teroxilon, Testolactone, Thiamidopamine, Thioguanine, Thiotepa, Thiazofurine, Tirapazamine,Toremifene citrate, triptolon acetate, triciribine phosphate, trimetrexate, trimetrexate glucuronide, triptorelin, tobradox hydrochloride, uramustine, uredepa, vapreotide, verteporfin, vinblastine sulfate, vincristine sulfate, vindesine, vindesine sulfate, vinposide sulfate, vinorelbine sulfate, vinorelbine tartrate, vinprodine sulfate, vinorelbine sulfate, vorozole, zeniplatin, jingsistatin, zorubicin hydrochloride. Other anticancer agents that can be used include, but are not limited to, 5-ethynyluracil, abiraterone, aclarubicin, acylfulvene, adecypenol, adolesin, aldesleukin, ALL-TK antagonists, hexamethylmelamine, aminostine, 2,4-dichlorophenoxyacetic acid (amidox), amifostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, andrographolide, angiogenesis inhibitors, antagonist D, antagonist G, anrelix, anti-dorsalization protein 1, antiandrogens, prostate cancer, antiestrogens, antineoplastons (antineoplasto n), antisense oligonucleotides, glycine aphidicolin, apoptosis gene regulators, apoptosis regulators, apurinic nucleic acids, ara-CDP-DL-PTBA, arginine deaminase, asulacrine, atamestane, amustine, axinastatin 1, axinastatin 2, axinastatin 3, azasetron, azatoxin, diazotyrosine, baccatin III derivatives, balanol, batimastat, BCR / ABL antagonists, benzochlorin, benzoylstaurosporine, beta-lactam derivatives, β-alethine, betaclamycin B, betulinic acid, bFGF inhibitors, bicalutamide, bisantrene, diazinon spermine, binefad, bistratene A, bizolexin, breflate, bropiridamine, budotitan, butylthionine imide, calcipotriol, calphostin C, camptothecin derivatives, canarypox IL2, capecitabine, carboxyamide-amino-triazole, carboxyamide triazole, CaRestM3, CARN700, cartilage Derived inhibitors, cardzelesin, casein kinase inhibitor (ICOS), castanospermine, bactericidal peptide B, cetrorelix, chlorins, chloroquinoxaline sulfonamide, cicaprost, cis-porphyrin, cladribine, enclomiphene analogs, clotrimazole, collismycin A, collismycin B, combretastatin A4, combretastatin analogs, conagenin, crambescidin 816, clenato, cryptophycin 8, cryptophycin A derivatives,Curacin A, cyclopentanthraquinone, cycloplatam, cypemycin, cytarabine sodium octadecyl phosphate, cytolytic factor, estradiol phosphate (cytostatin), dacximab, decitabine, dehydrogenated oxadiazine B, deslorelin, dexamethasone, right ifosfamide, right razoxane, right verapamil, diazadiquinone, dehydrogenated oxadiazine B, 3,4-dihydroxybenzohydroxamic acid (didox), diethylspermidine, dihydro 5-azacytidine, dihydropaclitaxel, 9-, dioxamycin, diphenylspiromustine, docetaxel, docosanol, dola Setron, doxifluridine, droloxifene, dronabinol, duocarmycin SA, ebselen, etiomustine, edilfosine, edrecolomab, eflornithine, elemene, etifur, epirubicin, etirostamide, estramustine analogs, estrogen agonists, estrogen antagonists, etanidazole, etoposide phosphate, exemestane, fadrozoles, fazarabine, fenretinide, filgrastim, finasteride, flavopiridol, fluasterone, fludarabine, fluorodaunorunicin, fomemax, formestane, fostramexine, fostramustine, gadolinium texap hyrin), gallium nitrate, galocitabine, ganirelix, gelatinase inhibitors, gemcitabine, glutathione inhibitors, hepsulfam, heregulin, cyclohexylbisacetamide, hyperanthrone, ibandronic acid, bilirubin, idoxifene, idemenone, imofosine, ilomastat, imidazoacridone, imiquimod, immunostimulatory peptides, insulin-like growth factor 1 receptor inhibitors, interferon agonists, interferons, interleukins, iodobenzylguanidine, iodoxorubicin, 4-sweet potato bitter alcohol, iropra, irsogladine, isobengazole, isohomohalicondrin B, itasetron, jasplakinolide , kahalalide F, lamellarin-N triacetic acid, lanreotide, leinamycin, lelagostine, mushroom polysaccharide sulfate, leptolstatin, letrozole, leukemia inhibitory factor, interferon alpha, leuprolide + estrogen + progesterone, leuprolide, levamisole, lirazo, linear polyamine analogs, lipophilic disaccharide peptides, lipophilic platinum compounds, lissoclinamide 7, lobaplatin, guanidineethylphosphoserine, lometrexol, lonidamine, losoxantrone, lovastatin, loxoribine, lutetiumtexaphyrin,Lysofylline, cytolytic peptide, metansine, mannostatin A, marimastat, maspin, mammary gland serine protease inhibitor, matrix lytic factor inhibitor, matrix metalloproteinase inhibitor, menolipin, thiobutanil, metirol, methionase, metoclopramide, MIF inhibitor, mifepristone, miltefosine, milistostatin, mismatched double-stranded RNA, mitoxantrone, dibromodole, mitomycin analogs, mitonadamide, mitotoxin, fibroblast growth factor-saponin, mitoxantrone, mofarotin, morasporastatin, monoclonal antibody, human chorionic gonadotropin, monophosphoryl lipid A+ mycobacterial cell wall skeleton, mopidarol, multidrug resistance gene inhibitor, multiple tumor suppressor 1-based therapy, mustard anticancer agent, Indian Ocean sponge (mycaperoxide) B, mycobacterial cell wall extract, myriaporone, N-acetyl danaline, N-substituted benzamide, nafarelin, nagrestip, naloxone + analgesic, napavin, naphterpin, natograstim, nedaplatin, nemorubicin, neridronic acid, neutral endopeptidase, nilutamide, nisamycin, nitric oxide regulator, nitrooxygen antioxidant, nitrullyn ), O6-benzylguanine, octreotide, okicenone, oligonucleotide, onapristone, ondansetron, ondansetron, oracin, oral cytokine inducer, ormaplatin, oxatron, oxaliplatin, oxaunomycin, paclitaxel, paclitaxel analog, paclitaxel derivative, palauamine, palmitoyl rhizoxin, pamidronate, panaxytriol, panomifen, parabactin, palzeptin, pegaspargase, perdecan, sodium pentothiosulfate, pentostatin, pentrozole, perflurane, perfosfamide, perillyl alcohol, phenazino mycin), phenylacetate, phosphatase inhibitors, picibanil, pilocarpine hydrochloride, pirarubicin, piroxine, placenta A, placenta B, plasminogen activator inhibitors, platinum complexes, platinum compounds, platinum-triamine complexes, porfibril sodium, porfibrin, chlorprednisolone, propyl diacridone, prostaglandin J2, proteasome inhibitors, protein A-based immunomodulators, protein kinase C inhibitors, protein kinase C inhibitors, microalgal, protein tyrosine phosphatase inhibitors, purine nucleoside phosphorylase inhibitors, purpurin, pyrazoline acridine, pyridoxalated hemoglobin polyoxyethylene conjugates,raf antagonists, raltitrexed, ramosetron, ras farnesyl protein transferase inhibitors, ras inhibitors, ras-GAP inhibitors, demethylated reteptine, etidronate rhenium Re186, rhizoxin, ribozymes, RII retinamide, rogulimide, rohitukine, romotide, roquimek, rubiginone B1, ruboxyl, safengo, saintopin, SarCNU, sarcophytol A, sargramostim, Sdi1 mimetics, semustine, senescence-derived inhibitor 1, sense oligonucleotides , signal transduction inhibitor, signal transduction regulator, single-chain antigen binding protein, sizoran, sobuzoxane, borca ​​sodium, sodium phenylacetate, solverol, somatomedin binding protein, sonarmin, fospartic acid, spicamycin D, spiromustine, splenopentin, spongistatin 1, squalamine, stem cell inhibitor, stem cell division inhibitor, stipiamide, matrix degrading inhibitor, sulfinosine, potent vasoactive intestinal peptide antagonist, suradista, suramin, swainsonine , synthetic mucopolysaccharides, tamustine, tamoxifen methyl iodide, tauromustine, tazarotene, tecogalan sodium, tegafur, tellurapyrylium, telomerase inhibitors, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, tetrazomine, thaliblastine, thiocobalamin, thrombopoietin, thrombopoietin mimetics, thymofasin, thymopoietin receptor agonist, thymosin, thyroid stimulating hormone, tin ethyl ester, tirapazamine, dichlorocyclopentadienyl titanium, topsentin, torel Mifen, totipotent stem cell factor, translation inhibitors, retinoic acid, triacetyl uridine, triciribine, trimetrexate, triptorelin, tropisetron, torosterol, tyrosine kinase inhibitors, tyrphostin, UBC inhibitors, ubenimex, urogenital sinus-derived growth inhibitory factor, ureakinase receptor antagonist, vapreotide, variolin B, vector system, erythrocyte gene therapy, vilaresol, veratrum, verdins, verteporfin, vinorelbine, vinxaltine, Vitaxin, vorozole, zanoteron, zeniplatin, benzyl retinol and statins.

[0154] In the fifth aspect of the present invention, a vaccine adjuvant is provided, which comprises the pyridine [3,2-d] pyrimidine-2-amine derivative or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative described in the first aspect, or consists of the same.

[0155] Specifically, the vaccine adjuvant also includes one or more pharmaceutically acceptable excipients, such as sodium chloride, phosphate, etc.

[0156] In the sixth aspect of the present invention, there is provided the use of the pyridine [3,2-d] pyrimidine-2-amine derivative or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative described in the first aspect in the preparation of a drug for preventing and / or treating diseases associated with TLR activity.

[0157] In some embodiments of the invention, the TLR is TLR7 and / or TLR8.

[0158] In the seventh aspect of the present invention, there is provided the use of the pyridine [3,2-d] pyrimidine-2-amine derivative or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative described in the first aspect in the preparation of a drug for preventing and / or treating diseases caused by or related to pathogen infection, immune diseases, inflammatory diseases, and tumors.

[0159] Specifically, the pathogen can be a microorganism, a parasite (protozoa, worms, etc.) or other medium; specifically, the above-mentioned microorganism can be selected from: one or more of: viruses, chlamydia, rickettsia, mycoplasma, bacteria, spirochetes, fungi, etc.

[0160] Specifically, examples of viruses include, but are not limited to, the following: human immunodeficiency virus (HIV), hepatitis virus (A, B, C, or D), herpes virus (such as VZV, HSV-I, HAV-6, HSV-II, CMV, Epstein-Barr virus), adenovirus, influenza virus, flavivirus, echovirus, rhinovirus, coxsackievirus, coronavirus (e.g., SARS-CoV, MERS and SARS-CoV-2), respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum virus, poliovirus, rabies virus, JC virus, arboviral encephalitis virus, rabies virus, filovirus (e.g., Ebola virus, Marburg virus); in particular HIV and HBV.

[0161] Specifically, examples of diseases caused by or associated with pathogen infection include, but are not limited to, viral hepatitis (such as hepatitis A, hepatitis B, hepatitis C, hepatitis D, etc.), AIDS, rabies, dengue fever, Ebola virus disease, etc.

[0162] In some embodiments of the present invention, the disease caused by pathogen infection or associated with pathogen infection is an infectious disease, in particular an infectious disease caused by viral infection, such as AIDS and hepatitis B.

[0163] In some embodiments of the present invention, the immune disease is an autoimmune disease, including but not limited to systemic lupus erythematosus, rheumatoid arthritis, inflammatory bowel disease, Sjogren's syndrome, polymyositis, vasculitis, Wegener's granulomatosis, sarcoidosis, ankylosing spondylitis, Reiter's syndrome, psoriatic arthritis, Behcet's syndrome, etc.

[0164] In some embodiments of the present invention, the inflammatory disease includes, but is not limited to, one or more of osteoarthritis, gout, chronic obstructive pulmonary disease, periodic fever, rash, lymphadenopathy, sepsis, inflammatory bowel disease, asthma, and allergies.

[0165] Specifically, tumors include, but are not limited to, human sarcomas and carcinomas, such as fibrosarcoma, myosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, pancreatic cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms tumor, cervical cancer, testicular tumor, lung cancer, small cell lung cancer, epithelial carcinoma, glioma leukemias, such as acute lymphocytic leukemia and acute myeloblastic leukemia (myeloblastic, promyelocytic, myelomonocytic, monocytic, and erythroblastic leukemias); chronic leukemias (chronic myeloid (granulocytic) leukemia and chronic lymphocytic leukemia); and polycythemia vera, lymphomas (Hodgkin's and non-Hodgkin's diseases), multiple myeloma, Waldenstrom's macroglobulinemia, and heavy chain disease.

[0166] In some embodiments of the present invention, the tumor is selected from the group consisting of: colon cancer, bladder cancer, melanoma, meningioma, lung cancer, pancreatic cancer.

[0167] In the eighth aspect of the present invention, there is provided the use of the pyridine [3,2-d] pyrimidine-2-amine derivative or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative described in the first aspect in the preparation of a drug for improving the body's positive immune response, enhancing the anti-HIV effect, improving the efficacy of immunotherapy, and preventing HIV rebound.

[0168] In the ninth aspect of the present invention, a method for selectively regulating (enhancing) TLR activity is provided, which comprises the step of administering the pyridine [3,2-d] pyrimidine-2-amine derivative described in the first aspect or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative or the pharmaceutical composition described in the fourth aspect.

[0169] In some embodiments of the invention, the TLR is TLR7 and / or TLR8.

[0170] In particular, the method is performed in vivo or in vitro.

[0171] In the tenth aspect of the present invention, a method for preventing and / or treating a disease associated with TLR activity is provided, comprising the step of administering to a subject in need thereof the pyridine [3,2-d] pyrimidine-2-amine derivative described in the first aspect or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative thereof or the pharmaceutical composition described in the fourth aspect.

[0172] In some embodiments of the invention, the TLR is TLR7 and / or TLR8.

[0173] In some embodiments of the present invention, the disease is selected from: diseases caused by pathogen infection or associated with pathogen infection, immune diseases, inflammatory diseases, tumors, as described in the seventh aspect of the present invention.

[0174] Specifically, the subject is a mammal, especially a human.

[0175] In the eleventh aspect of the present invention, a method for preventing and / or treating diseases caused by or associated with pathogen infection, immune diseases, inflammatory diseases, and tumors is provided, which comprises the step of administering to a subject in need thereof the pyridine [3,2-d] pyrimidine-2-amine derivative described in the first aspect or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative or the pharmaceutical composition described in the fourth aspect.

[0176] Specifically, the disease has the definition of the seventh aspect of the present invention.

[0177] Specifically, the subject is a mammal, especially a human.

[0178] In the twelfth aspect of the present invention, a method for enhancing the body's positive immune response is provided, comprising the step of administering to a subject in need thereof the pyridine [3,2-d] pyrimidine-2-amine derivative described in the first aspect or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative, the pharmaceutical composition described in the fourth aspect or the vaccine adjuvant described in the fifth aspect.

[0179] In the thirteenth aspect of the present invention, a method for enhancing the anti-HIV effect is provided, which comprises the step of administering to a subject in need thereof the pyridine [3,2-d] pyrimidine-2-amine derivative or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative described in the first aspect or the pharmaceutical composition described in the fourth aspect.

[0180] In the fourteenth aspect of the present invention, a method for improving the efficacy of immunotherapy is provided, which comprises the step of administering to a subject in need thereof the pyridine [3,2-d] pyrimidine-2-amine derivative described in the first aspect or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative or the pharmaceutical composition described in the fourth aspect.

[0181] In some embodiments of the present invention, the method further comprises infusing adoptive immunotherapy cells (eg, chimeric antigen receptor T cells) into the subject simultaneously with or subsequent to the above-mentioned administering step.

[0182] In the fifteenth aspect of the present invention, a method for preventing HIV rebound is provided, which comprises the step of administering to a subject in need thereof the pyridine [3,2-d] pyrimidine-2-amine derivative described in the first aspect or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative or the pharmaceutical composition described in the fourth aspect.

[0183] Specifically, the pyridine [3,2-d] pyrimidine-2-amine derivative or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative thereof described in the first aspect or the pharmaceutical composition described in the fourth aspect is used as a delayed reversal agent (LRA).

[0184] The inventors previously designed and prepared compound N01 (see Chinese patent CN108069963B). Due to its strong lipid solubility and alkalinity, it has potential cardiac safety risks. After a series of optimizations based on selectivity, PK / PD properties and safety, the inventors obtained the compound of the present invention, which has better physicochemical properties, drugability, cardiac safety and oral bioavailability, and will have very good research and application value in the medical field. DETAILED DESCRIPTION

[0185] Unless otherwise defined, all scientific and technical terms used in the present invention have the same meanings as commonly understood by one of ordinary skill in the art to which the present invention relates.

[0186] Related definitions:

[0187] The term "alkyl" refers to a hydrocarbon group formed by losing a hydrogen atom from an alkane molecule, which can be a straight chain or a branched chain and is connected to the rest of the molecule by a single bond. In the present invention, C0 alkyl refers to H, i.e., C0-C6 alkyl includes H and C1-C6 alkyl.

[0188] The term "alkylene" refers to a hydrocarbon group (divalent alkyl) formed by losing two hydrogen atoms from an alkane molecule, which can be a straight chain or a branched chain and is connected to the rest of the molecule by a single bond. In the present invention, C0 alkylene refers to a single bond, i.e., C0-C6 alkylene includes a single bond and C1-C6 alkylene.

[0189] The term "cycloalkylene" refers to a hydrocarbon group (a divalent group) formed by losing two hydrogen atoms from a cycloalkane molecule and connected to the rest of the molecule by a single bond. For example, cyclopropylene can be

[0190] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0191] The term "haloalkyl" refers to a group formed by replacing one or more hydrogen atoms in an alkyl group with a halogen atom (e.g., fluorine, chlorine, bromine or iodine), for example, -CHF2, -CH2F, -CF3, -CH2-CH2F, -CH2-CHF2, -CH2-CF3, -CH2CH2-CF3.

[0192] The term "heterocyclyl" refers to a 3- to 18-membered ring group containing 2 to 17 carbon atoms and 1 to 10 heteroatoms selected from N, O or S atoms. The heterocyclyl may be a monocyclic, bicyclic, tricyclic, or tetracyclic polycyclic system, which may include a fused (two rings share two ring atoms), a spiro (two rings share one ring atom) or a bridged (two rings share three or more ring atoms) ring system (excluding the case of linked rings). The heterocyclyl may be partially saturated (heteroaryl) or fully saturated (heterocycloalkyl). Suitable heteroaryl groups in the compounds of the present invention contain 1, 2 or 3 heteroatoms selected from N, O or S atoms, and include, for example, coumarin, including 8-coumarin, quinolyl, including 8-quinolyl, isoquinolyl, pyridyl, pyrazinyl, pyrazolyl, pyrimidinyl, furanyl, pyrrolyl, thienyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, imidazolyl, indolyl, isoindole, indazolyl, indolizinyl, phthalazinyl, pteridinyl, Purinyl, oxadiazolyl, thiadiazolyl, furazinyl, pyridazinyl, triazinyl, cinnolinyl, benzimidazolyl, benzofuranyl, benzofurazinyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, furopyridinyl, pyridooxazepinyl, pyridoazepinyl, pyridodiazepinyl, pyridooxepanyl, pyridocycloheptanyl, benzoxepanyl, benzoazepinyl, benzodiazepinyl, benzoxepanyl. Suitable heterocycloalkyl groups (i.e., saturated heterocyclic groups) in the compounds of the present invention contain 1, 2 or 3 heteroatoms selected from N, O or S atoms, and the heterocycloalkyl groups include, for example, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, oxathiolanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxirane, thiirane, azepaninyl, oxetan ... azepinyl, diazepinyl, triazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, dihydroindolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothiolanyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3H-indolyl and quinolizinyl.

[0193] The term "TLR" refers to Toll like receptor.

[0194] The term "pharmaceutically acceptable" refers to a material that has no adverse biological or other effects, for example, the material can be incorporated into a pharmaceutical composition administered to a patient without causing any adverse biological effects or interacting in a deleterious manner with any other ingredient in the composition in which it is contained. When the term "pharmaceutically acceptable" is used to refer to a pharmaceutical carrier or excipient, it means that the carrier or excipient meets the required standards of toxicology and manufacturing testing or is included in the inactive ingredient guidelines established by the U.S. Food and Drug Administration.

[0195] The term "pharmaceutically acceptable salt" refers to an acidic or basic salt that is theoretically non-toxic, non-irritating, and non-allergenic, and can achieve or provide clinically acceptable pharmacokinetic properties, absorption, distribution, and metabolic properties of the drug molecule, and can achieve the desired purpose. The salts described in the present invention include acidic or basic salts that are pharmaceutically acceptable to the acidic group, basic group, or amphoteric group of the compound. A list of suitable salts can be found in SM Birge, et al., J. Pharm. Sci., 66, 1-19 (1977).

[0196] The term "combination" or "pharmaceutical combination" means a product resulting from the mixing or combining of more than one active ingredient and includes both fixed and non-fixed combinations of the active ingredients.

[0197] The term "vaccine adjuvant" refers to a class of substances that can non-specifically change or enhance the body's specific immune response to antigens and play an auxiliary role.

[0198] The term "adoptive immune cell therapy" refers to a tumor treatment method that directly kills tumors or stimulates the body's anti-tumor immune response by re-infusing immune effector cells with anti-tumor activity that have been cultured and expanded in vitro. It mainly includes several major categories, such as TIL, LAK, CIK, DC, NK, TCR-T, and CAR-T.

[0199] The term "TLR disease" or "disease or disorder associated with TLR activity" refers to any disease state associated with a toll-like receptor.

[0200] The term "tumor" refers to an abnormal mass of tissue, wherein the growth of the mass exceeds the growth of normal tissue and is not coordinated with the growth of normal tissue. Tumors can be "benign" or "malignant", depending on the following characteristics: degree of cell differentiation (including morphology and function), growth rate, local invasion and metastasis. "Benign tumors" are usually well-differentiated, characterized by slower growth than malignant tumors, and remain confined to the site of origin. In addition, benign tumors do not have the ability to infiltrate, invade or metastasize to distant sites. In some cases, some "benign" tumors may later lead to malignant tumors, which may be caused by additional genetic changes in a subpopulation of the neoplastic cells of the tumor, and these tumors are called "precancerous tumors". "Malignant tumors" are usually poorly differentiated (anaplastic) and have a characteristic rapid growth, accompanied by progressive infiltration, invasion and destruction of surrounding tissues. In addition, malignant tumors usually have the ability to metastasize to distant sites.

[0201] The term "cancer" refers to a malignant tumor (Stedman's Medical Dictionary, 25th ed.; Hensyled.; Williams & Wilkins: Philadelphia, 1990).

[0202] The term "autoimmune disease" refers to a disease caused by the body's immune response to its own antigens, resulting in damage to its own tissues.

[0203] The term "inflammation" is the body's defense response to stimulation, manifested as redness, swelling, heat, pain, and dysfunction, etc. It can be infectious inflammation caused by infection, or non-infectious inflammation not caused by infection, such as inflammation caused by immune response (such as various types of hypersensitivity reactions, inflammation caused by some autoimmune diseases). The term "inflammatory disease" refers to a disease with inflammation.

[0204] The term "diseases associated with pathogen infection" mainly refers to diseases caused by pathogen infection, which include symptoms of body damage and infection response caused by pathogen invasion.

[0205] The term "patient" or "subject" and the like are used interchangeably herein and refer to any animal or cell thereof treated according to the methods described herein, whether in vitro or in situ. Specifically, the aforementioned animal includes mammals, e.g., rats, mice, guinea pigs, rabbits, dogs, monkeys (e.g., cynomolgus monkeys) or humans, in particular humans.

[0206] The term "treating" refers to preventing, curing, reversing, attenuating, alleviating, minimizing, inhibiting, suppressing and / or halting one or more clinical symptoms of a disease after onset of the disease.

[0207] The term "prevent" refers to avoiding, minimizing or making the onset or development of a disease difficult by treating it before it occurs.

[0208] The room temperature described in the present invention is 20-30°C.

[0209]

[00136] Various publications, patents, and published patent specifications are cited herein, the disclosures of which are incorporated by reference in their entireties.

[0210] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0211] Compound Preparation Example:

[0212] Example 1: Synthesis of intermediates

[0213]

[0214] (1) Synthesis of Compound M3

[0215] Dissolve clopyralid (50 g, 260.4 mmol) in N,N-dimethylformamide (800 mL), add guanidine hydrochloride (25 g, 263.2 mmol), cuprous oxide (7.4 g, 52.1 mmol) and cesium carbonate (211.6 g, 651.1 mmol), and stir at 120°C for 18 hours. Cool to room temperature, filter, fully dissolve the filter cake in methanol (1 L), filter, and concentrate the filtrate under reduced pressure to obtain the crude target product M3 as a gray-green solid (50 g, crude product). The product is directly used for the next step without purification. MS m / z (ESI): 197.0 [M+1].

[0216] (2) Synthesis of Compound M5

[0217] M3 (10 g, 51.0 mmol) was dissolved in N, N-dimethylformamide (80 mL), and N, N-diisopropylethylamine (19.7 g, 153.0 mmol), 1H-benzotriazol-1-yloxytripyrrolidino hexafluorophosphate (26.5 g, 51.0 mmol) and 2-methoxyethylamine (3.8 g, 51.0 mmol) were added and stirred at room temperature for 18 hours. The reaction solution was diluted with ethyl acetate (500 mL), washed with saturated brine (500 mL × 3), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: ethyl acetate (0.5% ammonia water)-petroleum ether gradient: 0-80%) to obtain the target product M5 as a yellow solid (1.1 g, 8.5%). MS m / z (ESI): 254.1 [M+1].

[0218] (3) Synthesis of Compound M7

[0219] Dissolve methyl 4-bromomethylbenzoate (2.7 g, 11.8 mmol) in tetrahydrofuran (80 mL), add zinc powder (2.3 g, 35.4 mmol) under ice bath, replace with nitrogen three times, and stir at 0 ° C nitrogen for 0.5 hours. Then add bis(triphenylphosphine)nickel chloride (780 mg, 1.2 mmol) and M5 (1.5 g, 5.9 mmol) in tetrahydrofuran (80 mL) under 0 ° C nitrogen, and stir at room temperature nitrogen for 18 hours. The reaction solution was quenched with water (200 mL), extracted with ethyl acetate (200 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: methanol-dichloromethane gradient: 0-3%) to obtain the target product M7 as a yellow solid (1.1 g, 50.8%). MS m / z (ESI): 368.2 [M+1].

[0220] (4) Synthesis of Compound M8

[0221] M7 (1.1 g, 3.0 mmol) was dissolved in tetrahydrofuran (40 mL), and a tetrahydrofuran solution of lithium aluminum hydroxide (1 M) (4.5 mL, 4.5 mmol) was slowly added under nitrogen at 0°C, and stirred for 2 hours under nitrogen at 0°C. The reaction solution was added dropwise to an iced aqueous solution of ammonium chloride (200 mL) to quench, and extracted with ethyl acetate (200 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: methanol-dichloromethane gradient: 0-4%) to obtain the target product M8 as a yellow oil (0.8 g, 78.7%). MS m / z (ESI): 340.1 [M+1].

[0222] (5) Synthesis of Compound M9

[0223] M8 (0.6 g, 1.8 mmol) was dissolved in dichloromethane (20 mL), thionyl chloride (1.1 g, 9 mmol) was slowly added at 0°C, and stirred at 0°C for 1 hour. The reaction solution was directly concentrated under reduced pressure to obtain the crude target product M9 as a brown solid (0.6 g, crude product), which was directly used for the next step without purification. MS m / z (ESI): 358.2 [M+1].

[0224] Example 2: Synthesis of Compound T1

[0225]

[0226] M9 (40 mg, 0.112 mmol) (prepared in Example 1) was dissolved in acetonitrile (5 mL), and N-methylpiperazine (16 mg, 0.168 mmol), potassium carbonate (32 mg, 0.224 mmol) and sodium iodide (1.6 mg, 0.011 mmol) were added and stirred at 60°C for 2 hours. After cooling to room temperature, the reaction solution was quenched with water (20 mL), extracted with ethyl acetate (20 mL×3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by C18 silica gel column chromatography (mobile phase: acetonitrile-water (0.05% formic acid) gradient: 5%-60%) to obtain the target product T1 as a white solid (12.7 mg, 27.1%). MS m / z (ESI): 422.2 [M+1]. 1 H NMR(400MHz,MeOD)δ8.49(s,2H),7.70(d,J=8.6Hz,1H),7.59(d,J=8.6Hz,1H),7.32(s,4H),4.26(s,2 H),3.88(t,J=5.5Hz,2H),3.70(t,J=5.5Hz,2H),3.63(s,2H),3.44(s,3H),3.03(s,4H),2.69(s,7H).

[0227] Example 3: Synthesis of Compound T2

[0228]

[0229] M9 (50 mg, 0.14 mmol) was dissolved in acetonitrile (2 mL), and N-hydroxyethylpiperazine (26.8 mg, 0.21 mmol), potassium carbonate (38.8 mg, 0.28 mmol) and sodium iodide (2.5 mg, 0.014 mmol) were added, and stirred at 60 ° C for 2 hours. After cooling to room temperature, the reaction solution was quenched with water (20 mL), extracted with ethyl acetate (20 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by C18 silica gel column chromatography (mobile phase: acetonitrile-water (0.05% formic acid) gradient: 5%-60%) to obtain the target product T2 as a white solid (16.7 mg, 26.4%). MS m / z (ESI): 452.2 [M+1]. 1 HNMR (400MHz, DMSO) δ7.81(s,1H),7.51(d,J=8.6Hz,1H),7.40(d,J=8.6Hz,1H),7.23(q,J=8.1Hz,4H),4.12(s,2H),3.66(dd,J=11.6,5.8 Hz,2H),3.56(t,J=5.8Hz,2H),3.49(t,J=6.1Hz,2H),3.42(s,2H),3.30(s,3H),2.50(d,J=1.7Hz,4H),2.46(t,J=6.2Hz,2H),2.38(s,4H).

[0230] Example 4: Synthesis of Compound T3

[0231]

[0232] (1) Synthesis of Compound T3-1

[0233] M9 (120 mg, 0.335 mmol) was dissolved in acetonitrile (5 mL), and tert-butyl piperazine-1-carboxylate (72 mg, 0.402 mmol), potassium carbonate (90 mg, 0.670 mmol) and sodium iodide (5 mg, 0.033 mmol) were added, and stirred at 60 ° C for 4 hours. After cooling to room temperature, the reaction solution was quenched with water (20 mL), extracted with ethyl acetate (20 mL × 3), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with an eluent system (mobile phase: methanol-dichloromethane gradient: 0-7%) to obtain the target product T3-1 as a yellow solid (132 mg, 77.6%). MS m / z (ESI): 508.3 [M+1].

[0234] (2) Synthesis of Compound T3

[0235] T3-1 (132 mg, 0.260 mmol) was dissolved in a methanol solution of hydrogen chloride (6 M) (10 mL) and stirred at 60°C for 2 hours. After cooling to room temperature, the reaction solution was concentrated under reduced pressure, and the residue was purified by C18 silica gel column chromatography (mobile phase: acetonitrile-water (0.05% formic acid) gradient: 5%-60%) to obtain the target product T3 as a yellow solid (36 mg, 34.1%). MS m / z (ESI): 408.3 [M+1]. 1 H NMR (400MHz, MeOD) δ7.78(d,J=8.6Hz,1H),7.70(d,J=8.6Hz,1H),7.61(d,J=7.9Hz,2H),7.51(d,J=8.0H z,2H),4.47(s,2H),4.34(s,2H),3.91(t,J=5.5Hz,2H),3.71(t,J=5.5Hz,2H),3.62(s,8H),3.44(s,3H).

[0236] Example 5: Synthesis of Compound T4

[0237]

[0238] M9 (60 mg, 0.17 mmol) was dissolved in acetonitrile (4 mL), 4-aminotetrahydropyran (26 mg, 0.26 mmol), potassium carbonate (94 mg, 0.68 mmol) and sodium iodide (26 mg, 0.17 mmol) were added, and stirred at 60 ° C for 2 hours. After cooling to room temperature, the reaction solution was quenched with water (50 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by C18 silica gel column chromatography (mobile phase: acetonitrile-water (0.05% formic acid) gradient: 5%-15%) to obtain the target product T4 as a yellow solid (25.9 mg, 36.4%). MS m / z (ESI): 423.0 [M+1]. 1 H NMR (400MHz, CD3OD) δ8.48(s,2H),7.72(d,J=8.6Hz,1H),7.60(d,J=8.6Hz,1H),7.51–7.39(m,4H),4.31(s,2H),4.21(s,2H),4.05(dd,J=11 .7,4.5Hz,2H),3.88(t,J=5.5Hz,2H),3.70(t,J=5.5Hz,2H),3.51–3.37(m,6H),2.09(dd,J=12.3,2.3Hz,2H),1.72(td,J=12.2,4.6Hz,2H).

[0239] Example 6: Synthesis of Compound T5

[0240]

[0241] To a solution of M9 (100 mg, 0.280 mmol) in ACN (2 mL) was added K2CO3 (116 mg, 0.841 mmol), compound T5-1 (53.4 mg, 0.336 mmol) and NaI (4.2 mg, 0.028 mmol). The mixture was stirred at 80°C for 2 hours. The reactant was evaporated and purified by C18 gel chromatography (5-75% ACN / H2O mobile phase system, containing 0.05% formic acid) to obtain the target product T5 as a white solid (19.8 mg, 14.7% yield).

[0242] LC / MS:B230007-041-P1-35150.D(ES,m / z):[M+H] + =481.2.

[0243] 1 H NMR(400MHz,MeOD)δ7.74(d,J=8.6Hz,1H),7.66(d,J=8.6Hz,1H),7.44–7.3 6(m,4H),4.29(s,2H),3.99–3.92(m,3H),3.89(t,J=5.5Hz,2H),3.70(d,J=5 .5Hz,2H),3.69–3.58(m,2H),3.42(s,3H),3.05(dd,J=27.6,11.8Hz,2H),2. 65(td,J=12.0,3.5Hz,1H),2.47–2.29(m,3H),1.71(dt,J=14.5,7.6Hz,2H).

[0244] Example 7: Synthesis of Compound T6

[0245]

[0246] (1) Synthesis of Compound T6-3

[0247] To a solution of T6-1 (1 g, 10.1 mmol) in ACN (10 mL) was added DIEA (5.73 g, 44.4 mmol), KI (123 mg, 0.741 mmol) and T6-2 (1.48 g, 8.916 mmol). The mixture was stirred at room temperature for 18 hours. The reaction was quenched with H2O (300 mL) and extracted with EA (300 mL×2). The organic layer was evaporated and purified by silica gel chromatography (0-8% MeOH / DCM) to give the target product T6-3 as a yellow oil (220 mg, 11.8% yield).

[0248] LC / MS:B230007-035-3P1-34131.D(ES,m / z):[M+H] + =186.1.

[0249] (2) Synthesis of Compound T6-4

[0250] To a solution of T6-3 (200 mg, 0.560 mmol) in ACN (2 mL) was added K2CO3 (232 mg, 1.682 mmol), M9 (124.4 mg, 0.672 mmol) and KI (10 mg, 0.060 mmol). The reaction was quenched with H2O (50 mL) and extracted with EA (50 mL×2). The organic layer was evaporated and purified by silica gel chromatography (0-5% MeOH / DCM) to give the target product T6-4 as a yellow solid (70 mg, 24.7% yield).

[0251] LC / MS:B230007-037-2P1-34787.D(ES,m / z):[M+H] + =507.2.

[0252] (3) Synthesis of Compound T6

[0253] To a solution of T6-4 (70 mg, 0.138 mmol) in MeOH (1 mL) was added H2O (1 mL) and LiOH (29.1 mg, 0.693 mmol). The mixture was stirred at room temperature for 2 hours. The reaction was evaporated and purified by C18 gel chromatography (5-75% ACN / H2O mobile phase system, containing 0.05% formic acid) to give the target product T6 as a white solid (15.8 mg, 23.9% yield).

[0254] LC / MS:B230007-039-P1-39228.D(ES,m / z):[M+H] + =479.2.

[0255] 1H NMR (400MHz, DMSO) δ7.56(s,1H),7.52(d,J=8.6Hz,1H),7.39(d,J=8.6Hz,1H),7.25(q,J=8.1Hz,4H),6.07(s,2H),4 .14(s,2H),3.90(s,2H),3.70–3.66(m,2H),3.59(t,J=5.7Hz,2H),3.48(d,J=9.9Hz,2H),3.34(s,2H),3.33(s,3H).

[0256] Example 8: Synthesis of Compound T7

[0257]

[0258] (1) Synthesis of Compound T7-2

[0259] 3-Methoxy-4-bromomethylbenzoic acid methyl ester (T7-1) (300 mg, 1.18 mmol) was dissolved in tetrahydrofuran (8 mL), and zinc powder (230 mg, 3.54 mmol) was added under ice bath, and nitrogen was replaced three times, and stirred at 0 ° C nitrogen for 0.5 hours. Then, bis(triphenylphosphine)nickel chloride (78 mg, 0.12 mmol) and M5 (150 mg, 0.59 mmol) (prepared in Example 1) in tetrahydrofuran (8 mL) were added under 0 ° C nitrogen, and stirred at room temperature nitrogen for 18 hours. The reaction solution was quenched with water (50 mL), extracted with ethyl acetate (50 mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: methanol-dichloromethane gradient: 0-3%) to obtain the target product T7-2 as a yellow solid (230 mg, 48.8%). MS m / z(ESI):398.2[M+1].

[0260] (2) Synthesis of Compound T7-3

[0261] T7-2 (220 mg, 0.554 mmol) was dissolved in tetrahydrofuran (10 mL), and a tetrahydrofuran solution of lithium aluminum hydroxide (1 M) (0.8 mL, 0.8 mmol) was slowly added under nitrogen at 0°C, and stirred for 2 hours under nitrogen at 0°C. The reaction solution was added dropwise to an iced aqueous solution of ammonium chloride (50 mL) to quench, and extracted with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: methanol (1% ammonia water)-dichloromethane gradient: 0-5%) to obtain the target product T7-3 as a yellow oil (127 mg, 59.4%). MS m / z (ESI): 370.1 [M+1].

[0262] (3) Synthesis of Compound T7-4

[0263] T7-3 (120 mg, 0.324 mmol) was dissolved in dichloromethane (5 mL), thionyl chloride (114 mg, 0.973 mmol) was slowly added at 0°C, and stirred at 0°C for 1 hour. The reaction solution was directly concentrated under reduced pressure to obtain the crude target product T7-4 as a brown solid (120 mg, crude product), which was directly used for the next step without purification. MS m / z (ESI): 388.2 [M+1].

[0264] (4) Synthesis of Compound T7

[0265] T7-4 (65 mg, 0.168 mmol) was dissolved in acetonitrile (3 mL), and morpholine (26 mg, 0.251 mmol), potassium carbonate (46 mg, 0.336 mmol) and sodium iodide (2.6 mg, 0.017 mmol) were added, and stirred at 60°C for 2 hours. After cooling to room temperature, the reaction solution was quenched with water (20 mL), extracted with ethyl acetate (20 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by C18 silica gel column chromatography (mobile phase: acetonitrile-water (0.05% formic acid) gradient: 5%-60%) to obtain the target product T7 as a white solid (13.4 mg, 18.2%). MS m / z (ESI): 439.2 [M+1]. 1 H NMR (400MHz, MeOD) δ8.37(s,1H),7.56(d,J=8.7Hz,1H),7.41(d,J=8.7Hz,1H),7.09(d,J=7.6Hz,1H),6.93(s,1H),6.81(d,J=7.5Hz ,1H),4.12(s,2H),3.75(t,J=5.5Hz,2H),3.72(s,3H),3.61(s,4H),3.57(t,J=5.6Hz,4H),3.31(s,3H),3.21(dt,J=3.2,1.6Hz,4H).

[0266] Example 9: Synthesis of Compound T8

[0267]

[0268] T7-4 (60 mg, 0.155 mmol) was dissolved in acetonitrile (2 mL), and dimethylamine tetrahydrofuran solution (2M) (0.25 mL, 0.465 mmol), potassium carbonate (43 mg, 0.309 mmol) and sodium iodide (3 mg, 0.016 mmol) were added, and stirred at 50 ° C for 2 hours. After cooling to room temperature, the reaction solution was quenched with water (20 mL), extracted with ethyl acetate (20 mL × 3), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by C18 silica gel column chromatography (mobile phase: acetonitrile-water (0.05% formic acid) gradient: 5%-60%) to obtain the target product T8 as a white solid (10.9 mg, 17.8%). MS m / z (ESI): 397.2 [M+1]. 1 H NMR (400MHz, MeOD) δ8.54(s,1H),7.65–7.60(m,1H),7.48(d,J=8.6Hz,1H),7.28(d,J=7.6Hz,1H),7.09(s,1H),6.99(d,J=7.6Hz,1H ),4.25(s,2H),4.03(d,J=8.8Hz,2H),3.87(s,3H),3.83(t,J=5.4Hz,2H),3.68(t,J=5.5Hz,2H),3.43(s,3H),2.67(d,J=5.2Hz,6H).

[0269] Example 10: Synthesis of Compound T9

[0270]

[0271] (1) Synthesis of Compound T9-1

[0272] M7 (200 mg, 0.54 mmol) (prepared in Example 1) was dissolved in methanol (4 mL), lithium hydroxide (40 mg, 1.65 mmol) and water (2 mL) were added, and stirred at room temperature for 4 hours. The reaction solution was adjusted to pH 2 with a methanol solution of hydrochloric acid and then directly concentrated under reduced pressure to obtain a crude target product T9-1 (240 mg, crude product) as a light yellow solid. The product was directly used for the next step without purification. MS m / z (ESI): 354.1 [M+1].

[0273] (2) Synthesis of Compound T9

[0274] T9-1 (80 mg, 0.23 mmol) was dissolved in N,N-dimethylformamide (3 mL), and N,N-diisopropylethylamine (88.5 mg, 0.69 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (86.9 mg, 0.23 mmol) and dimethylamine (2M) (0.11 mL) were added and stirred at room temperature for 1 hour. The reaction solution was quenched with water (50 mL), extracted with ethyl acetate (50 mL×3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: dichloromethane-methanol gradient: 0-10%) to obtain the target product T9 as a yellow solid (22.2 mg, 28%). MS m / z (ESI): 381.3 [M+1]. 1 HNMR (400MHz, DMSO) δ8.15(s,1H),7.73(t,J=5.5Hz,1H),7.48(d,J=8.6Hz,1H),7.39(d,J=8.6Hz,1H),7.30(t,J=6.0H z,4H),6.38(s,2H),4.14(s,2H),3.62(dd,J=11.5,5.8Hz,2H),3.52(t,J=5.7Hz,2H),3.26(s,3H),2.93–2.84(m,6H).

[0275] Example 11: Synthesis of Compound T10

[0276]

[0277] (1) Synthesis of Compound T10-2

[0278] To a solution of T9-1 (100 mg, 0.283 mmol) in DMF (1 mL) were added DIEA (87.7 mg, 0.679 mmol), HATU (86.1 mg, 0.226 mmol) and T10-1 (28.3 mg, 0.318 mmol). The mixture was stirred at room temperature for 2 hours. The reaction was quenched with H2O (20 mL) and extracted with EA (20 mL×2). The organic layer was evaporated to give the target product T10-2 as a yellow solid (50 mg, crude).

[0279] LC / MS:B230007-008-4P1-23153.D(ES,m / z):[M+H] + =425.2.

[0280] (2) Synthesis of Compound T10

[0281] To a solution of T10-2 (45 mg, 0.106 mmol) in MeOH (1 mL) was added H2O (1 ml) and LiOH (22.3 mg, 0.531 mmol). The mixture was stirred at room temperature for 1 hour. The reaction was evaporated and purified by C18 gel chromatography (5-75% ACN / H2O mobile phase system, containing 0.05% formic acid) to give the desired product T10 as a white solid (9.2 mg, 21.1% yield).

[0282] LC / MS:B230007-009-P1-23798.D(ES,m / z):[M+H] + =411.2.

[0283] 1 H NMR (400MHz, DMSO) δ8.70(t,J=5.8Hz,1H),7.79(dd,J=10.8,7.1Hz,3H),7.51(d,J=8.6Hz,1H),7.41(dd,J=8.4,1.5 Hz,3H),6.47(s,2H),4.20(s,2H),3.87(d,J=5.8Hz,2H),3.66(q,J=5.8Hz,2H),3.56(t,J=5.8Hz,2H),3.30(s,3H).

[0284] Example 12: Synthesis of Compound T11

[0285]

[0286] (1) Synthesis of Compound T11-2

[0287] To a solution of T11-1 (50 mg, 0.314 mmol) in MeOH (1 mL) was added SOCl2 (186 g, 1.576 mmol). The mixture was stirred at room temperature for 1 hour. The reaction was evaporated to give the target product T11-2 as a yellow oil (50 mg, crude).

[0288] LC / MS:B230007-010-1P1-23384.D(ES,m / z):[M+H] + =174.2.

[0289] (2) Synthesis of Compound T11-3

[0290] To a solution of T9-1 (100 mg, 0.283 mmol) in DMF (1 mL) was added DIEA (109.6 mg, 0.850 mmol), HATU (107.6 mg, 0.283 mmol) and T11-2 (49 mg, 0.283 mmol). The mixture was stirred at room temperature for 2 hours. The reaction was quenched with ice-H2O (20 mL) and extracted with EA (20 mL×3). The organic layer was evaporated to give the target product T11-3 as a yellow solid (50 mg, crude).

[0291] LC / MS:B230007-011-3P1-23945.D(ES,m / z):[M+H] + =509.2.

[0292] (3) Synthesis of Compound T11

[0293] To a solution of T11-3 (45 mg, 0.089 mmol) in MeOH (1 mL) were added H2O (0.5 mL) and LiOH (18.6 mg, 0.443 mmol). The mixture was stirred at room temperature for 1 hour. The reaction was quenched with ice-H2O (20 mL) and extracted with EA (20 mL×2). The organic layer was evaporated and purified by C18 gel chromatography (5-75% ACN / H2O mobile phase system, containing 0.05% formic acid) to obtain the target product T11 as a white solid (17.1 mg, 39.0% yield).

[0294] LC / MS:B230007-012-P1-24142.D(ES,m / z):[M+H] + =495.2.

[0295] 1 H NMR(400MHz,MeOD)δ8.39(s,1H),7.60(d,J=8.6Hz,1H),7.49(d,J=8.6Hz,1H),7 .32(d,J=8.2Hz,2H),7.26(d,J=8.2Hz,2H),4.33(d,J=12.3Hz,1H),4.18(s,2H), 3.95–3.63(m,3H),3.57(t,J=5.5Hz,2H),3.42(d,J=46.8Hz,2H),3.21(dt,J=3. 2,1.6Hz,3H),3.17–2.51(m,3H),2.24(d,J=47.0Hz,2H),1.61(d,J=79.3Hz,2H).

[0296] Biological test examples:

[0297] Example 1: EC of small molecule agonists 50 Determination

[0298] Reagent: HEK-Blue TM hTLR7、HEK-Blue TM Reagents required for hTLR8 and control HEK-Blue Null2k cell culture and detection: DMEM (4.5g / l glucose), bovine serum FBS, streptomycin (50μg / ml), penicillin (50U / ml) Blasticidin (10mg / ml), ZeocinTM (10mg / ml), Normocin (50mg / ml), HEK-BlueTMDetection. Basic culture medium: DMEM + 10% FBS + streptomycin + penicillin + Normocin (100μg / ml); Selective culture medium for resistance genes: DMEM + 10% FBS + streptomycin + penicillin + Normocin (100μg / ml) + Blasticidin (100μg / ml) and Zeocin TM (100 μg / ml).

[0299] Cell preparation: HEK-Blue TM hTLR7、HEK-Blue TM hTLR8 cell line was purchased from Invivogen. Take out the cells frozen in liquid nitrogen, quickly put them into a 37℃ water bath, shake them from time to time, and completely thaw them within 1 minute; transfer the cells to 15mL preheated basal culture medium and resuspend them; centrifuge at 1000r / min for 5min, discard the supernatant; resuspend in 1ml basal culture medium, transfer to a T25 culture flask, add 5ml culture medium, and culture in a 37℃ incubator; after two stable passages, add selective antibiotics for screening: HEK-Blue TM hTLR7 or HEK-Blue TM hTLR8: 100 μg / ml Zeocin+Blasticidin (30 μg / ml), HEK-Blue Null2-k: 100 μg / ml Zeocin; change the medium three times a week; when the cell volume reaches 70%-80%, add PBS and gently blow to make the cells fall off.

[0300] Activity detection steps: (1) Add 20 μL of the sample to be tested to each well of a 96-well plate (set up three replicates); (2) Add 20 μL of the positive control (R848); Add 20 μL of the negative control (ddH2O); (3) Remove the T75 culture flask from the incubator, discard the culture medium, add 5-10 mL of preheated PBS, and gently blow the cells; add 2-5 mL of PBS to the culture flask and return it to the incubator for 1-2 minutes, gently blow to scatter the cells; (4) Mix well, count the cells, and do not centrifuge. Use HEK-Blue TM Resuspend the cells in the test solution to adjust the number of cells. Do not incubate for too long to avoid too dark background or false positives. The number of HEK-Blue hTLR cells is about 2.2×10 5 / mL, 180 μL (about 40,000 cells) was added to each well, and the amount of HEK-Blue Null2-k cells was about 2.8×10 5 / ml, 180 μL per well (about 50,000 cells); (5) incubate at 37°C for 6-16 h, and detect SEAP readings at 620-655 nm.

[0301] Effect % = (mean OD of drug group at each concentration - mean OD of H2O group) / (OD of positive drug group max Average value-average value of OD in H2O group)×100. GraphPad Prism5 software was used to fit the Lg [concentration]-effect curve to calculate the EC 50 .

[0302] Experimental results: Concentration for 50% of maximal effect (EC 50 ) refers to the concentration of small molecules that can cause 50% of the maximum agonist effect. The experimental results are shown in Table 1. The half-maximal effect concentration of the positive control R848 is hTLR7 (75nM) and hTLR8 (480nM). In Table 1, the unit of agonist activity is nanomolar unless otherwise specified. The results show that the test compound has good inhibitory activity against hTLR7 and hTLR8.

[0303] Table 1 EC of compounds 50 Measurement results

[0304]

[0305]

[0306]

[0307] Example 2: Investigation of the physicochemical properties of the compounds

[0308] Test compounds: compounds T2, T3, T7, and control compound N01 (for its preparation, see Chinese patent CN108069963B).

[0309] The clogP (lipid-water partition coefficient) values ​​of the test compounds were predicted by chemdraw software, and the results are shown in Table 2, indicating that the compounds of the present invention have better hydrophilicity.

[0310] The solubility of the compounds in phosphate buffer at pH = 7.4 was further tested. The results are shown in Table 2. The solubility of compounds T2, T3, and T7 was increased by more than 10 times compared with compound N01.

[0311] The above results indicate that the compounds of the present invention have better physical and chemical properties and drugability.

[0312] Table 2 Lipid-water partition coefficient and solubility test results of the compounds

[0313] Compound clogP Solubility (μM) N01 4.7 125 T2 2.9 1670 T3 3.0 1458 T7 2.9 1276

[0314] Example 3: In vitro metabolic stability study of compounds

[0315] Evaluate the metabolic stability of compounds in in vitro liver microsomes. The experimental operation method and steps are as follows:

[0316] 1 μM of the compound was incubated in 1 mg / mL liver microsomes at 37°C. The reaction system was 0.1 M potassium phosphate buffer (pH 7.4), containing 3 mmol / L MgCl2 and 1 mmol / L NADPH. At the beginning of the reaction, the small molecule and liver microsomes were pre-incubated at 37°C for 10 minutes, and then the cofactor NADPH was added to initiate the metabolic reaction, and the reaction was terminated with three volumes of ice methanol containing internal standard (200 nM labetalol) at 0, 5, 10, 20, 30 and 60 minutes. After incubation at room temperature for 10 minutes, the sample was centrifuged at 4000 rpm and 4°C for 20 minutes, and the supernatant was diluted 1:1 with chromatographic pure water for quantitative analysis. The quantification of the compound was performed by LC-MS / MS on an Agilent quadrupole time-of-flight instrument (Agilent 6510). The peak of the designated ion signal was detected after 6 minutes of separation on a C18 column (50×2.1mm; 2.6μm) in a mobile phase gradient containing 0.1% formic acid-water. For metabolite identification, the mobile phase gradient was 1:1, the retention time was extended to 20 minutes, and the metabolites were analyzed by MS / MS, and the obvious molecular ion peaks were analyzed. The detailed operation process is as follows:

[0317] 1) Add 10 μL / well of compound working solution to all 96-well reaction plates (T0, T5, T10, T20, T30, T60) except the blank.

[0318] 2) Add 80 μL / well of the microsome solution to all reaction wells (T0, T5, T10, T20, T30, T60, and blank).

[0319] 3) Pre-incubate at 37°C for 10 minutes.

[0320] 4) After pre-incubation, add 10 μL / well of NADPH to each reaction plate to start the reaction.

[0321] 5) At the set end time point, 300 μL / well of 4° C. methanol solution (containing internal standard: 200 nM labetalol) was added to each reaction plate to terminate the reaction.

[0322] 6) Seal each plate and shake for 10 minutes.

[0323] 7) After shaking, centrifuge at 4000 rpm and 4°C for 20 minutes.

[0324] 8) Take out 300 μl of supernatant, add 300 μl / well of HPLC-grade water, mix well, and then detect by LC-MS.

[0325] Data Analysis:

[0326] The first-order kinetic equation was used to calculate the metabolic half-life (T1 / 2) and metabolic clearance rate (CLint(mic)) (μL / min / mg).

[0327] First-order kinetic formula:

[0328] C t =C0 exp(-k e t), k e is a constant

[0329] When C t =1 / 2C0, T 1 / 2 =Ln2 / k e =0.693 / k e ;

[0330] C Lint (mic)=0.693 / T 1 / 2 ×1 / (Microsome concentration:mg / mL)

[0331] The results are shown in Table 3.

[0332] Table 3 Results of liver microsomal stability evaluation of compounds

[0333]

[0334] The test results showed that the half-life of the new compound T3 and T2 was significantly extended by 5 to 25 times. This indicates that the oxygen substitution of the butyl chain slowed down the metabolic rate, and it is speculated that the oxidation of the β-position of the N01 alkyl chain is a potential metabolic pathway. The above results indicate that the new compound may have better oral bioavailability.

[0335] Example 4: Investigation of cardiac potassium channel (hERG) inhibitory activity of compounds

[0336] The rapidly activated potassium channel encoded by the human ether-a-go-go-related gene (hERG) is an important ion channel involved in the formation of phase 3 repolarization of the cardiac action potential. Drug blockade of the hERG channel can lead to prolonged cardiac repolarization, which is manifested as a prolonged QT interval on the electrocardiogram, known as long QT syndrome. Drug-induced delayed ventricular repolarization may in some cases induce a fatal arrhythmia, torsade de pointes. In this study, the whole-cell patch clamp technique (a standard method for hERG safety evaluation) was used to study the inhibitory effect of small molecules on the hERG potassium channel and to assess their risk of inducing ventricular repolarization toxicity.

[0337] The test results are shown in Table 4. Compound N01 showed strong inhibitory activity against hERG, with an inhibition rate of about 63.22% at a concentration of 3 μM, which is at the edge of the clinical drug safety requirements. Compared with N01, the overall lipophilicity of compounds T2 and T3 was significantly reduced, and the inhibitory activity against hERG was also greatly reduced. The inhibition rates at 10 μM were 30.09% and 32.32%, respectively, which are weak inhibitors according to the general standard (IC 50 >10μM). The test results show that the compounds of the present invention significantly improve cardiac safety.

[0338] Table 4 hERG inhibitory activity evaluation results of compounds

[0339]

[0340] a IC 50 value.

[0341] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

[0342] The aforementioned embodiments and methods described in the present invention may be varied based on the ability, experience and preference of those skilled in the art.

[0343] In the present invention, merely listing the steps of the method in a certain order does not constitute any limitation on the order of the method steps.

Claims

1. A compound or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or isotope derivative thereof, wherein the compound has the following structure: in, L1 is C1-C6 alkylene or C3-C6 cycloalkylene; L2 is C1-C6 alkylene or -C(O)-; Ring A is selected from: R0 is selected from: H, C1-C6 alkyl, C1-C6 haloalkyl, -O(C0-C6 alkyl), halogen, cyano, -N(C0-C6 alkyl)(C0-C6 alkyl), heterocyclyl; R1 is selected from: H, -O(C0-C6 alkyl), -(C1-C6 alkylene)-OH, -OC(O)(C0-C6 alkyl), and a basic tertiary amine group; R2 and R3 are independently selected from: H, C1-C6 alkyl, C1-C6 haloalkyl, -(C1-C6 alkylene)-COO-(C0-C6 alkyl), 4-7 membered heterocyclyl (especially 5- or 6-membered saturated heterocyclyl, such as nitrogen-containing heterocyclyl or oxygen-containing heterocyclyl); or R2 and R3 together with the nitrogen atom to which they are commonly attached form a heterocycle, which is optionally substituted by one or more groups selected from the following: halogen, C1-C6 alkyl, C1-C6 haloalkyl, -(C0-C6 alkylene)-OH, -(C0-C6 alkylene)- -(C0-C6 alkyl)-COO-(C0-C6 alkyl), -(C0-C6 alkylene)-N(C0-C6 alkyl)(C0-C6 alkyl), -(C0-C6 alkylene)-heterocyclyl; wherein the heterocyclyl is optionally substituted by one or more groups selected from the following: halogen, C1-C6 alkyl, C1-C6 haloalkyl, -(C0-C6 alkylene)-OH, -(C0-C6 alkylene)-COO-(C0-C6 alkyl), -(C0-C6 alkylene)-N(C0-C6 alkyl)(C0-C6 alkyl); Preferably, the compound has the following structure:

2. The compound according to claim 1, characterized in that L1 is selected from:

3. The compound according to claim 1, characterized in that L2 is selected from:

4. The compound according to claim 1, characterized in that R1 is H or -O(C1-C3 alkyl), preferably H or methoxy.

5. The compound according to any one of claims 1 to 4, characterized in that R2 is C1-C3 alkyl (e.g. methyl), R3 is C1-C3 alkyl (e.g. methyl); or, R2 is selected from: H, C1-C3 alkyl (e.g. methyl), C1-C3 haloalkyl (e.g. -CF3, -CH2CF3), R3 is -(C1-C3 alkylene)-COO-(C0-C3 alkyl) (e.g. or, R2 is H, R3 is a 4-6 membered saturated heterocyclic group (e.g. R2 and R3 together with the nitrogen atom to which they are attached form a 4-10 membered saturated heterocyclic ring (e.g. The heterocycle is optionally substituted by one or more groups selected from the group consisting of halogen (e.g., F), C1-C3 alkyl (e.g., methyl), C1-C3 haloalkyl (e.g., trifluoromethyl), -(C0-C3 alkylene)-OH (e.g., -OH, -(C0-C3 alkylene)-COO-(C0-C3 alkyl) (e.g. -COOH, -(C0-C6 alkylene)-N(C0-C6 alkyl)(C0-C6 alkyl)(e.g. -NH2, -(C0-C6 alkylene)-(4-10 membered saturated heterocyclic group), wherein the 4-10 membered saturated heterocyclic group (e.g. Optionally substituted by one or more groups selected from the following: halogen, C1-C3 alkyl, C1-C3 haloalkyl, -(C0-C3 alkylene)-OH, -(C0-C3 alkylene)-COO-(C0-C3 alkyl), -(C0-C3 alkylene)-N(C0-C3 alkyl)(C0-C3 alkyl).

6. The compound according to any one of claims 1 to 4, characterized in that The compound has the following structure: in, X is selected from: single bond, N, O, CH, C(O), S, S(O), S(O)2; R4 is one or more independent substituents on the ring, which are independently selected from: H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -(C0-C6 alkylene)-OH, -(C0-C6 alkylene)-COO-(C0-C6 alkyl), -(C0-C6 alkylene)-N(C0-C6 alkyl)(C0-C6 alkyl), -(C0-C6 alkylene)-(4-10 membered saturated heterocyclyl), wherein the 4-10 membered saturated heterocyclyl is optionally substituted with one or more groups selected from the following: halogen, C1-C3 alkyl, C1-C3 haloalkyl, -(C0-C3 alkylene)-OH, -(C0-C3 alkylene)-COO-(C0-C3 alkyl), -(C0-C3 alkylene)-N(C0-C3 alkyl)(C0-C3 alkyl); Preferably, the compound has the following structure: Wherein, R4 is selected from: H, C1-C6 alkyl, -(C1-C6 alkylene)-OH, -(C1-C6 alkylene)-COO-(C0-C6 alkyl); or, The compound has the following structure: Wherein, R4 is one or more independent substituents on the ring, which are independently selected from: H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -(C0-C6 alkylene)-OH, -(C0-C6 alkylene)-COO-(C0-C6 alkyl), -(C0-C6 alkylene)-N(C0-C6 alkyl)(C0-C6 alkyl), -(C0-C6 alkylene)-(4-10 membered saturated heterocyclyl), wherein the 4-10 membered saturated heterocyclyl is optionally substituted by one or more groups selected from the following: halogen, C1-C3 alkyl, C1-C3 haloalkyl, -(C0-C3 alkylene)-OH, -(C0-C3 alkylene)-COO-(C0-C3 alkyl), -(C0-C3 alkylene)-N(C0-C3 alkyl)(C0-C3 alkyl).

7. The compound according to claim 1, characterized in that The compound is selected from the following structures:

8. The compound according to claim 1, characterized in that The stereoisomers have the following structures:

9. A pharmaceutical composition comprising the pyridinium [3, 2-d] pyrimidin-2-amine derivative according to any one of claims 1 to 8 or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative thereof, and one or more pharmaceutically acceptable excipients.

10. A vaccine adjuvant comprising the pyridine [3, 2-d] pyrimidine-2-amine derivative according to any one of claims 1 to 8 or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative thereof, and optionally, one or more pharmaceutically acceptable excipients.

11. Use of the pyridine [3,2-d] pyrimidine-2-amine derivative according to any one of claims 1 to 8 or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative in the preparation of a drug for preventing and / or treating a disease associated with TLR activity; Preferably, the TLR is TLR7 and / or TLR8.

12. Use of the pyridine [3,2-d] pyrimidine-2-amine derivative according to any one of claims 1 to 8 or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative in the preparation of a drug for preventing and / or treating diseases caused by or related to pathogen infection, immune diseases, inflammatory diseases, and tumors; Preferably, the pathogen is a virus selected from the group consisting of human immunodeficiency virus (HIV), hepatitis virus (A, B, C, or D), herpes virus (such as VZV, HSV-I, HAV-6, HSV-II, CMV, Epstein-Barr virus), adenovirus, influenza virus, flavivirus, echovirus, rhinovirus, coxsackievirus, coronavirus (e.g., SARS-CoV, MERS and SARS-CoV-2), respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum virus, poliovirus, rabies virus, JC virus, arboviral encephalitis virus, rabies virus, filovirus (e.g., Ebola virus, Marburg virus); More preferably, the disease caused by pathogen infection or associated with pathogen infection is selected from: viral hepatitis (such as hepatitis A, hepatitis B, hepatitis C, hepatitis D, etc.), AIDS, rabies, dengue fever, Ebola virus disease; Preferably, the immune disease is an autoimmune disease selected from the group consisting of systemic lupus erythematosus, rheumatoid arthritis, inflammatory bowel disease, Sjogren's syndrome, polymyositis, vasculitis, Wegener's granulomatosis, sarcoidosis, ankylosing spondylitis, Reiter's syndrome, psoriatic arthritis, and Behcet's syndrome; Preferably, the inflammatory disease is selected from one or more of osteoarthritis, gout, chronic obstructive pulmonary disease, periodic fever, rash, lymphadenopathy, sepsis, inflammatory bowel disease, asthma, and allergy; Preferably, the tumor is selected from the group consisting of fibrosarcoma, myosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, pancreatic cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung cancer, small cell lung cancer, epithelial carcinoma, glioma, astrocytoma, One or more of medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, leukemia (e.g., acute lymphocytic leukemia and acute myeloblastic leukemia (myeloblastic, promyelocytic, myelomonocytic, monocytic and erythroblastic leukemia); chronic leukemia (chronic myeloid (granulocytic) leukemia and chronic lymphocytic leukemia)), polycythemia vera, lymphoma (Hodgkin's disease and non-Hodgkin's disease), multiple myeloma, Waldenstrom's macroglobulinemia and heavy chain disease.

13. Use of the pyridine [3,2-d] pyrimidine-2-amine derivative according to any one of claims 1 to 8 or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative in the preparation of a drug for improving the body's positive immune response, enhancing the anti-HIV effect, improving the efficacy of immunotherapy, and preventing HIV rebound.

14. A method for selectively regulating (enhancing) TLR activity, comprising the step of administering the pyridine [3,2-d] pyrimidine-2-amine derivative according to any one of claims 1 to 8 or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, isotope derivative thereof, or the pharmaceutical composition according to the fourth aspect; Preferably, the TLR is TLR7 and / or TLR8; Preferably, the method is performed in vivo or in vitro.

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