Aminopyridine compounds and their applications

Abstract

Aminopyridine compounds, specifically compounds represented by formula (P) and pharmaceutically acceptable salts thereof, can be used to prepare drugs for treating deep fungal infections. #imgabs0#

Description

[0001] The present invention claims the following priority:

[0002] CN202011482610.1, application date: December 15, 2020;

[0003] CN202110528944.6, application date: May 14, 2021. Technical Field

[0004] The present invention relates to a series of aminopyridyl compounds, in particular to compounds represented by formula (P) and pharmaceutically acceptable salts thereof. Background Art

[0005] Invasive fungal disease (IFD) is the most lethal type of fungal infection, with morbidity and mortality rates showing a dramatic upward trend. The fungal cell wall is primarily composed of glucan, chitin, and mannoproteins. Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are anchored to the cell membrane and cell wall, mediating cross-linking between mannoproteins and glucan, and significantly influencing fungal cell wall synthesis, adhesion, and morphological transformation. Gwt1 is a key acetylase in the GPI synthesis process and plays an important role in the formation of GPI precursors. Inhibiting Gwt1 activity blocks GPI-AP synthesis, preventing fungal surface mannoproteins from cross-linking to the cell wall. This, in turn, impairs the ability to adhere to host surfaces and cell wall integrity, exerting an antifungal effect. Summary of the Invention

[0006] The present invention provides a compound represented by formula (P) or a pharmaceutically acceptable salt thereof,

[0007]

[0008] in,

[0009] Ring A is selected from

[0010] T1 and T2 are selected from CH and N;

[0011] L1 is selected from -O-, -CH2O- and -OCH2-, wherein -CH2O- and -OCH2- are optionally substituted with 1 or 2 halogens;

[0012] Each R1 is independently selected from H, F, Cl, Br, I, OH and NH2;

[0013] Each R2 is independently selected from F, Cl, OH, NH2 and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 R a replace;

[0014] R3 is selected from

[0015] Each R4 is independently selected from H, F, Cl, Br and I;

[0016] Each R a are independently selected from F, Cl, Br and I;

[0017] m is selected from 1, 2, 3 and 4;

[0018] n is selected from 1, 2, 3 and 4;

[0019] z is selected from 1, 2 and 3.

[0020] In some embodiments of the present invention, L1 is selected from -O-, -CH2O- and -OCH2-, and other variables are as defined in the present invention.

[0021] In some embodiments of the present invention, T1 is selected from N, and other variables are as defined in the present invention.

[0022] In some embodiments of the present invention, T2 is selected from CH, and other variables are as defined herein.

[0023] In some embodiments of the present invention, each R1 is independently selected from H and F, and other variables are as defined in the present invention.

[0024] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0025] In some embodiments of the present invention, each R2 is independently selected from F, Cl, OH, NH2, CH3, CH2CH3 and CH(CH3)2, and the CH3, CH2CH3 and CH(CH3)2 are optionally replaced by 1, 2 or 3 R a Substitution, other variables are as defined in the present invention.

[0026] In some embodiments of the present invention, each R2 is independently selected from F, Cl, OH, NH2, CH3, CHF2 and CH2F, and other variables are as defined in the present invention.

[0027] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0028] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0029] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0030] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0031] In some embodiments of the present invention, the ring A is selected from Other variables are as defined in the present invention.

[0032] In some embodiments of the present invention, each R4 is independently selected from H and F, and other variables are as defined in the present invention.

[0033] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0034] In some embodiments of the present invention, the compound or a pharmaceutically acceptable salt thereof is selected from

[0035]

[0036] in,

[0037] L1, T1, T2, R1, R2, R4 and m are as defined herein.

[0038] The present invention provides a compound represented by formula (IV) or a pharmaceutically acceptable salt thereof,

[0039]

[0040] in,

[0041] L2 is selected from

[0042] T1 and T2 are selected from CH and N;

[0043] L1 is selected from -O-, -CH2O- and -OCH2-;

[0044] R1 is selected from H, F, Cl, Br, I, OH and NH2;

[0045] Each R2 is independently selected from F, Cl, OH, NH2 and C 1-3Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 R a replace;

[0046] Each R a are independently selected from F, Cl, Br and I;

[0047] m is selected from 1, 2, 3 and 4;

[0048] Wherein formula (III) does not include molecules

[0049] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0050] In some embodiments of the present invention, each R2 is independently selected from F, Cl, OH, NH2, CH3, CH2CH3 and CH(CH3)2, and the CH3, CH2CH3 and CH(CH3)2 are optionally replaced by 1, 2 or 3 R a Substitution, other variables are as defined in the present invention.

[0051] In some embodiments of the present invention, each R2 is independently selected from F, NH2, CH3, CHF2 and CH2F, and other variables are as defined in the present invention.

[0052] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0053] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0054] In some embodiments of the present invention, the compound or a pharmaceutically acceptable salt thereof is selected from

[0055]

[0056] in,

[0057] Ar is selected from

[0058] L1, T1, T2, R1, R2 and m are as defined in the present invention;

[0059] And formula (I) does not include molecules

[0060] The present invention provides a compound represented by formula (III) or a pharmaceutically acceptable salt thereof,

[0061]

[0062] in,

[0063] L2 is selected from

[0064] T1 and T2 are selected from CH and N;

[0065] L1 is selected from -O-, -CH2O- and -OCH2-;

[0066] R1 is selected from H, F, Cl, Br, I, OH and NH2;

[0067] Wherein formula (III) does not include molecules

[0068] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0069] In some embodiments of the present invention, the compound or a pharmaceutically acceptable salt thereof is selected from

[0070]

[0071] in,

[0072] Ar is selected from

[0073] L1, T1, T2 and R1 are as defined in the present invention;

[0074] And formula (I) does not include molecules

[0075] Some other solutions of the present invention are obtained by any combination of the above variables.

[0076] The present invention also provides the following compounds or pharmaceutically acceptable salts thereof:

[0077]

[0078]

[0079] The present invention also provides use of the compound or a pharmaceutically acceptable salt thereof in preparing a drug for treating diseases related to Gwt1.

[0080] Technical Effects

[0081] The compound of the present invention has good antibacterial activity against Candida, Cryptococcus, and Aspergillus, can combat mouse mortality caused by candidemia, and has excellent therapeutic effects on Candida vaginal infection. Furthermore, the compound of the present invention has excellent pharmacokinetic properties.

[0082] Related definitions

[0083] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as indefinite or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0084] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0085] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; and salts of amino acids (such as arginine) and organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups and can be converted into either base or acid addition salts.

[0086] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of the two.

[0087] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

[0088] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0089] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability to rotate freely about double bonds or single bonds forming ring carbon atoms.

[0090] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0091] Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

[0092] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed bond Indicate the relative configuration of stereocenters with a wavy line Indicates a wedge-shaped solid key or dotted wedge key Or use a wavy line Indicates a straight solid bond and straight dashed bond

[0093] Unless otherwise specified, when a compound contains a double bond structure, such as a carbon-carbon double bond, a carbon-nitrogen double bond, or a nitrogen-nitrogen double bond, and each atom on the double bond is connected to two different substituents (in a double bond containing a nitrogen atom, a lone pair of electrons on the nitrogen atom is considered as a substituent to which it is connected), if a wavy line is used between the atom on the double bond and its substituent in the compound, When connected, it represents the (Z) isomer, (E) isomer or a mixture of the two isomers of the compound. For example, the following formula (A) represents that the compound exists in the form of a single isomer of formula (A-1) or formula (A-2) or in the form of a mixture of two isomers of formula (A-1) and formula (A-2); the following formula (B) represents that the compound exists in the form of a single isomer of formula (B-1) or formula (B-2) or in the form of a mixture of two isomers of formula (B-1) and formula (B-2). The following formula (C) represents that the compound exists in the form of a single isomer of formula (C-1) or formula (C-2) or in the form of a mixture of two isomers of formula (C-1) and formula (C-2).

[0094]

[0095]

[0096] Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium at room temperature and readily interconvert into each other. If tautomerism is possible (e.g., in solution), chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via reorganization of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0097] Unless otherwise indicated, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of that isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0098] Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

[0099] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C). For example, deuterated drugs can be formed by replacing hydrogen with heavy hydrogen. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have advantages such as reduced toxic side effects, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of this invention.

[0100] The terms "optional" or "optionally" mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0101] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may be substituted or unsubstituted, and unless otherwise specified, the type and number of substituents can be any on the basis of chemical achievable.

[0102] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, with each occurrence of R being an independent choice. Furthermore, combinations of substituents and / or their variants are permissible only if such combinations result in stable compounds.

[0103] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0104] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0105] When the linking group is listed without specifying its linking direction, its linking direction is arbitrary, for example, The connecting group L is -MW-, in which case -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form You can also connect ring A and ring B in the opposite direction of reading from left to right to form Combinations of linkers, substituents, and / or variations thereof are permissible only if such combinations result in stable compounds.

[0106] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the chemical bond connection mode is non-positional and there are H atoms at the connectable sites, when the chemical bond is connected, the number of H atoms at the site will decrease accordingly with the number of connected chemical bonds, and become a group with a corresponding valence. The chemical bond connecting the site to other groups can be a straight solid bond. Straight dotted key or wavy lines For example, the straight solid bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy line in the phenyl group indicates that it is connected to other groups through the carbon atoms at positions 1 and 2 in the phenyl group; Indicates that any linkable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least In these four connection methods, even if an H atom is drawn on -N-, Still includes For groups connected in this way, when one chemical bond is connected, the H at that site will be reduced by one and become a corresponding monovalent piperidine group.

[0107] Unless otherwise specified, the term “C 1-6 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. 1-6 Alkyl groups include C 1-5 、C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-4 , C6 and C5 alkyl, etc.; which can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-6 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, and the like.

[0108] Unless otherwise specified, the term “C 1-4 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 4 carbon atoms. 1-4 Alkyl groups include C 1-2 、C 1-3 and C 2-3 Alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-4 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), and the like.

[0109] Unless otherwise specified, the term “C 1-3 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

[0110] Unless otherwise specified, C n-n+m or C n -C n+m Any specific case including n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C10 、C 11 , and C 12 , also includes any range from n to n+m, such as C 1-12 Including C 1-3 、C 1-6 、C 1-9 、C 3-6 、C 3-9 、C 3-12 、C 6-9 、C 6-12 , and C 9-12 Similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, an 11-membered ring, and a 12-membered ring, and also includes any range from n to n+m, for example, a 3-12-membered ring includes a 3-6-membered ring, a 3-9-membered ring, a 5-6-membered ring, a 5-7-membered ring, a 6-7-membered ring, a 6-8-membered ring, and a 6-10-membered ring, etc.

[0111] The term "protecting group" includes, but is not limited to, an "amino protecting group," a "hydroxy protecting group," or a "thiol protecting group." The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to, formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert-butyloxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-bis-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like. The term "hydroxy protecting group" refers to a protecting group suitable for preventing side reactions at the hydroxyl group. Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl and tert-butyl; acyl groups such as alkanoyl (e.g., acetyl); arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like.

[0112] The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention.

[0113] The structures of the compounds of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compounds, the absolute configuration can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data on the cultured single crystal using a Bruker D8 venture diffractometer, using CuKα radiation as the light source and scanning mode: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration. DETAILED DESCRIPTION

[0114] The present invention is described in detail below by way of examples, but is not intended to limit the present invention in any way. While the present invention has been described in detail herein, and specific embodiments thereof have been disclosed, it will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0115] Example 3

[0116] Synthesis route:

[0117]

[0118] Step 1: Synthesis of compound WX003-2

[0119] Compound WX003-1 (100 mg, 314.10 μmol) and compound WX001-1 (73 mg, 314.10 μmol) were dissolved in acetonitrile (5 mL). Cesium carbonate (256 mg, 785.26 μmol), XPhos (45 mg, 94.23 μmol), and Pd(CH3CN)2Cl2 (8 mg, 31.41 μmol) were added. The reaction was stirred at 90°C under nitrogen for 2 hours. LCMS showed that the starting materials were completely reacted. The reaction solution was directly filtered, and the filtrate was concentrated. The crude product was separated and purified by preparative thin-layer chromatography (petroleum ether:ethyl acetate = 1:1) to obtain compound WX003-2. MS m / z (ESI): 516.3 [M+H] + .

[0120] Step 2: Synthesis of compound WX003

[0121] Compound WX003-2 (50 mg, 96.97 μmol) was dissolved in formic acid (233 mg, 4.85 mmol) and stirred at 15°C for 16 hours. The reaction mixture was concentrated, and the crude product was purified by preparative HPLC (formic acid system, column: Phenomenex Luna C18, 75×30 mm×3 μm; mobile phase: [water (0.2% formic acid)-acetonitrile]; acetonitrile percentage: 20%-50%, 8 min). Compound WX003 was obtained. MS m / z (ESI): 316.1 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ = 8.17-8.16 (m, 1H), 7.92-7.90 (m, 1H), 7.73-7.70 (m, 1H), 7.49-7.47 (m, 1H), 7.47-7. 42(m,4H),6.99-6.97(m,1H),6.86(d,J=8.4Hz,1H),6.52-6.49(m,1H),6.08(s,2H),5.33(s,2H),3.92(s,2H).

[0122] Example 4

[0123] Synthesis route:

[0124]

[0125] Step 1: Synthesis of compound WX004-1

[0126] WX001-1 (7 g, 29.95 mmol) and Cs2CO3 (19.52 g, 59.91 mmol) were dissolved in THF (210 mL). After nitrogen purge, Pd(MeCN)2Cl2 (777 mg, 3.00 mmol) and XPhos (2.14 g, 4.49 mmol) were added. The atmosphere was purged with nitrogen again, and trimethylsilyl acetylene (14.71 g, 149.77 mmol, 20.75 mL) was added via syringe. The reaction mixture was heated to 65°C and stirred for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (0-10% ethyl acetate in petroleum ether) to afford WX004-1. 1 H NMR (400MHz, CDCl3) δ=8.21-8.19(m,1H),7.65-7.60(m,1H),7.46-7.42(m,2H),7.40-7. 36(m,2H),6.92-6.90(m,1H),6.84-6.81(m,1H),5.39(s,2H),3.68(s,2H),0.21(s,9H).

[0127] Step 2: Synthesis of compound WX004-2

[0128] WX004-1 (0.9 g, 3.05 mmol) was dissolved in THF (12 mL), and HOAc (366 mg, 6.09 mmol, 348 μL) and TBAF (1 M THF, 6.09 mL) were added. The reaction was stirred at 20°C for 16 hours. Water (20 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated sodium chloride solution (10 mL × 3), and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (0-10% ethyl acetate in petroleum ether) to obtain WX004-2. 1 H NMR (400MHz, CDCl3) δ = 8.20 (dd, J = 1.3, 5.0Hz, 1H), 7.61-7.59 (m, 1H), 7.46-7.44 (m, 2H), 7.42-7.38 (m, 2H), 6.92-6.90 (m, 1H), 6.82 (d, J = 8.4Hz, 1H), 5.39 (s, 2H), 3.64 (d, J = 2.8H, 2H), 2.21 (t, J = 2.8Hz, 1H).

[0129] Step 3: Synthesis of compound WX004

[0130] To a reaction flask, WX004-3 (1 g, 4.55 mmol), WX004-2 (1.01 g, 4.55 mmol), and THF (100 mL) were added, followed by Pd(MeCN)2Cl2 (177 mg, 681.79 μmol), XPhos (325 mg, 681.79 μmol), CuI (87 mg, 454.52 μmol), and TEA (3.68 g, 36.36 mmol, 5.06 mL). The atmosphere was replaced with nitrogen three times, and the reaction mixture was stirred at 15°C for 16 hours. The mixture was filtered through celite, and the filtrate was added with water (50 mL) and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 1:1 to 0:1) and then by HPLC (Phenomenex Luna C18 column, 200 × 40 mm × 10 μm; mobile phase: [water (0.2% FA)-ACN]; ACN%: 15%-55%, 8 min) to obtain WX004. MS m / z (ESI): 316 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ = 8.17-8.16 (m, 1H), 7.99 (s, 1H), 7.72-7.71 (m, 1H), 7.42-7.36 (m, 5H), 7.0 0-6.98(m,1H),6.85(d,J=8.4Hz,1H),6.39(d,J=8.8Hz,1H),6.24(s,2H),5.32(s,2H),3.83(s,2H).

[0131] Example 5

[0132] Synthesis route:

[0133]

[0134] Compounds WX004-2 (563 mg, 2.52 mmol) and WX005-1 (500 mg, 2.10 mmol) were dissolved in THF (20 mL). Pd(MeCN)2Cl2 (54 mg, 210.08 μmol), XPhos (150 mg, 315.12 μmol), CuI (40 mg, 210.08 μmol), and TEA (1.70 g, 16.81 mmol, 2.34 mL) were added. The atmosphere was replaced with nitrogen three times and stirred at 15°C for 16 hours. The reaction mixture was filtered and the filtrate was concentrated to obtain the crude product. The crude product was initially isolated and purified by column chromatography (gradient elution, petroleum ether:ethyl acetate = 10:1 to 2:1), followed by preparative separation and purification (formic acid system, column: Phenomenex Luna C18 200×40mm×10μm; mobile phase: [water (0.2% FA)-ACN]; ACN %: 50%-90%, 8 min) to obtain WX005. MS m / z (ESI): 334 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ:8.17-8.16(m,1H),7.72-7.70(m,1H),7.53-7.51(m,1H),7.43-7.36(m,4H) ,6.99-6.98(m,1H),6.86(d,J=8.4Hz,1H),6.70(s,2H),6.30-6.27(m,1H),5.33(s,2H),3.86(s,2H).

[0135] Example 6

[0136] Synthesis route:

[0137]

[0138] To a reaction flask, WX006-1 (100 mg, 427.28 μmol) and WX004-2 (95 mg, 427.28 μmol) were added, followed by THF (5 mL) and stirring. Pd(MeCN)2Cl2 (11 mg, 42.73 μmol), XPhos (30 mg, 64.09 μmol), CuI (8 mg, 42.73 μmol), and TEA (346 mg, 3.42 mmol, 476 μL) were then added sequentially. The atmosphere was replaced with nitrogen three times, and the mixture was stirred at 15°C for 16 hours. The reaction mixture was filtered through celite and concentrated under reduced pressure to obtain the crude product. The crude product was separated by preparative thin-layer chromatography (petroleum ether:ethyl acetate = 2:1) and purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75 × 30 mm × 3 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; ACN%: 35%-55%, 8 min) to obtain WX006. MS m / z (ESI): 330 [M+H] + , 1 H NMR(400MHz, CDCl3)δ:8.19(d,J=3.2Hz,1H),7.58-7.61(m,1H),7.41-7.46(m,5H),6.87–6.90(m,1H) ), 6.81 (d, J = 8.4Hz, 1H), 6.29 (d, J = 8.4Hz, 1H), 5.40 (s, 2H), 4.51 (s, 2H), 3.86 (s, 2H), 2.52 (s, 3H).

[0139] Example 7

[0140] Synthesis route:

[0141]

[0142] Compound WX004-2 (191 mg, 854.57 μmol) and compound WX007-1 (0.2 g, 854.57 μmol) were dissolved in tetrahydrofuran (4 mL). Triethylamine (692 mg, 6.84 mmol, 952 μL) was added. After nitrogen purge, cuprous iodide (16 mg, 85.46 μmol), bis(acetonitrile)palladium dichloride (22 mg, 85.46 μmol), and 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (61 mg, 128.18 μmol) were added. The atmosphere was purged with nitrogen again and stirred at 40°C for 16 hours. The reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (eluent: 0-50% ethyl acetate in petroleum ether) to obtain compound WX007. MS m / z (ESI): 330.2 [M+H] + , 1H NMR (400MHz, CDCl3) δ = 8.23-8.17 (m,1H),8.08(s,1H),7.65-7.57(m,1H),7.49-7.41(m,4H),7.37(s,1H),6.94-6.89 (m,1H),6.82(d,J=8.3Hz,1H),5.39(s,2H),4.60(s,2H),3.84(s,2H),2.13(s,3H).

[0143] Example 8

[0144] Synthesis route:

[0145]

[0146] Compound WX008-1 (200 mg, 840.33 μmol) and compound WX004-2 (225 mg, 1.01 mmol) were dissolved in tetrahydrofuran (8 mL). Triethylamine (680 mg, 6.72 mmol) was added. After nitrogen purge, cuprous iodide (16 mg, 84.03 μmol), bis(acetonitrile)palladium dichloride (22 mg, 84.03 μmol), and 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (60 mg, 126.05 μmol) were added. The atmosphere was purged with nitrogen again and stirred at 40°C for 16 hours. The reaction mixture was cooled to room temperature, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75×30mm×3μm; mobile phase: [water (0.225% FA)-ACN]; ACN%: 50%-60%, 7 min) to obtain WX008. MS m / z (ESI): 334.0 [M+H] + , 1 H NMR (400MHz, CDCl3) δ = 8.11 (dd, J = 1.2, 5.0Hz, 1H), 7.53-7.50 (m, 1H), 7.40-7.36 (m, 2H), 7.34-7.31 (m, 2H), 7.21 (d, J = 1. 5Hz,1H),7.18(d,J=1.6Hz,1H),6.81(dd,J=5.6,6.8Hz,1H),6.73(d,J=8.2Hz,1H),5.30(s,2H),4.71(s,2H),3.74(s,2H).

[0147] Example 9

[0148] Synthesis route:

[0149]

[0150] Step 1: Synthesis of compound WX009-3

[0151] Compound WX009-1 (3.97 g, 26.07 mmol) and compound WX009-2 (3 g, 26.07 mmol, 2.38 mL) were dissolved in acetonitrile (30 mL), and K2CO3 (10.81 g, 78.21 mmol) was added. The mixture was stirred at 85°C for 21 hours. Water (30 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL x 2). The organic phases were combined and washed with saturated sodium chloride solution (20 mL). The organic phases were concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (0-15% ethyl acetate in petroleum ether) to obtain compound WX009-3. MS m / z (ESI): 247.8 [M+H] + , 1 H NMR (400MHz, CDCl3) δ=8.14-8.07(m,2H),7.83-7.79(m,1H),7.24-7.20(m,2H),6.84-6.82(m,1H),6.65-6.74(m,1H),3.94(s,3H).

[0152] Step 2: Synthesis of compound WX009-4

[0153] Compound WX009-3 (2 g, 8.09 mmol) was dissolved in toluene (30 mL). After nitrogen replacement, the system was cooled to 0°C and diisobutylaluminum hydride (1 M in THF, 24.27 mL) was slowly added dropwise. The reaction mixture was stirred for 2 hours. The reaction mixture was slowly added to potassium bisulfate solution (20 mL), stirred for 10 minutes, and extracted with ethyl acetate (20 mL x 3). The organic phases were combined and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (0-30% ethyl acetate in petroleum ether) to obtain compound WX009-4. MS m / z (ESI): 220.0 [M+H] + , 1 H NMR (400MHz, CDCl3) δ=7.76-7.72(m,1H),7.40(d,J=8.4Hz,2H),7.13(d,J=8.4Hz,2H),6.74-6.72(m,1H),6.63-6.61(m,1H),4.69(s,2H).

[0154] Step 3: Synthesis of compound WX009-5

[0155] Thionyl chloride (10 mL) was added to the reaction flask, the system was cooled to 0°C, and compound WX009-4 (1 g, 4.56 mmol) was added. The reaction mixture was warmed to 20°C and stirred for 30 minutes. The reaction mixture was concentrated under reduced pressure to obtain a crude product. Saturated sodium bicarbonate solution (50 mL) was added to the crude product, and the product was extracted with ethyl acetate (50 mL x 2). The organic phases were combined and concentrated under reduced pressure to obtain compound WX009-5. MS m / z (ESI): 238.0 [M+H] + , 1 H NMR (400MHz, CDCl3) δ = 7.70-7.68 (m, 1H), 7.39-7.32 (m, 2H), 7.09-7.04 (m, 2H), 6.68 (dd, J = 1.2, 8.0Hz, 1H), 6.55 (dd, J = 2.4, 7.6Hz, 1H), 4.54 (s, 2H).

[0156] Step 4: Synthesis of compound WX009-7

[0157] Compound WX009-5 (0.7 g, 2.95 mmol) was dissolved in tetrahydrofuran (15 mL), and cesium carbonate (1.92 g, 5.89 mmol) was added. After nitrogen purge, 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (211 mg, 441.81 μmol) and bis(acetonitrile)palladium dichloride (76 mg, 294.54 μmol) were added. The atmosphere was again purged with nitrogen, and trimethylsilyl acetylene (1.45 g, 14.73 mmol, 2.04 mL) was added by syringe. The reaction mixture was stirred at 65°C for 16 hours. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (0-10% ethyl acetate in petroleum ether) to obtain compound WX009-7. MS m / z (ESI): 300.1 [M+H] + , 1 H NMR (400MHz, CDCl3) δ = 7.80-7.75 (m, 1H), 7.40 (d, J = 8.8Hz, 2H), 7.14-7.10 (m, 2H), 6.74-6.73 (m, 1H), 6.62 (dd, J = 2.4, 7.6Hz, 1H), 3.69 (s, 2H), 0.21 (s, 9H).

[0158] Step 5: Synthesis of compound WX009-8

[0159] Compound WX009-7 (0.7 g, 2.34 mmol) was dissolved in tetrahydrofuran (10 mL), and acetic acid (281 mg, 4.68 mmol, 267 μL) and tetrabutylammonium fluoride (1 M solution in tetrahydrofuran) (1 M, 4.68 mL) were added. The reaction was stirred at 20°C for 16 hours. Water (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (0-10% ethyl acetate in petroleum ether). Compound WX009-8 was obtained. MS m / z (ESI): 228.0 [M+H] + , 1 H NMR (400MHz, CDCl3) δ = 7.79-7.73 (m, 1H), 7.41 (d, J = 8.4Hz, 2H), 7.13 (d, J = 8.4Hz, 2H ),6.75-6.73(m,1H),6.64-6.62(m,1H),3.65(d,J=2.8Hz,2H),2.23(t,J=2.8Hz,1H).

[0160] Step 6: Synthesis of compound WX009

[0161] Compound WX004-3 (0.5 g, 2.27 mmol) and compound WX009-8 (620 mg, 2.73 mmol) were dissolved in tetrahydrofuran (20 mL). Triethylamine (1.84 g, 18.18 mmol, 2.53 mL) was added. After nitrogen purge, cuprous iodide (43 mg, 227.26 μmol), bis(acetonitrile)palladium dichloride (59 mg, 227.26 μmol), and 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (163 mg, 340.89 μmol) were added. The atmosphere was again purged with nitrogen, and the reaction mixture was stirred at 40°C for 16 hours. After cooling to room temperature, the reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75×30 mm×3 μm; mobile phase: [water (0.225% FA)-ACN]; ACN%: 15%-45%, 7 min). Compound WX009 was obtained. MS m / z (ESI): 320.1 [M+H] + , 1H NMR (400MHz, CDCl3) δ = 8.14 (s, 1H), 7.78-7.76 (m, 1H), 7.54 (dd, J = 2.0, 8.8Hz, 1H), 7.44 (d, J = 8.8Hz, 2H), 7.17-7.1 0(m,2H),6.75(dd,J=1.2,8.0Hz,1H),6.62(dd,J=2.4,7.6Hz,1H),6.49(d,J=8.4Hz,1H),5.17(s,2H),3.84(s,2H).

[0162] Example 10

[0163] Synthesis route:

[0164]

[0165] Step 1: Synthesis of compound WX010-2

[0166] Compound WX010-1 (0.2 g, 1.59 mmol) was dissolved in acetic acid (2 mL). The system was cooled to 0°C and N-iodosuccinimide (357 mg, 1.59 mmol) was added. The reaction was stirred at 20°C for 2 hours. Water (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (0-30% ethyl acetate in petroleum ether). Compound WX010-2 was obtained. MS m / z (ESI): 252.7 [M+H] + .

[0167] Step 2: Synthesis of compound WX010

[0168] Compound WX010-2 (0.1 g, 396.78 μmol) and compound WX004-2 (106 mg, 476.14 μmol) were dissolved in tetrahydrofuran (4 mL). Triethylamine (321 mg, 3.17 mmol, 441.82 μL) was added. After nitrogen purge, cuprous iodide (8 mg, 39.68 μmol), bis(acetonitrile)palladium dichloride (10 mg, 39.68 μmol), and 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (28 mg, 59.52 μmol) were added. The atmosphere was again purged with nitrogen, and the reaction mixture was stirred at 40°C for 16 hours. The reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75×30 mm×3 μm; mobile phase: [water (0.225% FA)-ACN]; ACN%: 30%-60%, 7 min) to obtain compound WX010. MS m / z (ESI): 348.1 [M+H] + ,1 H NMR (400MHz, CDCl3) δ = 8.20 (d, J = 4.4Hz, 1H), 7.66-7.58 (m, 1H), 7.52-7.41 (m, 4H), 7.23 (d, J = 10.8H z,1H),6.95-6.89(m,1H),6.82(d,J=8.0Hz,1H),5.39(s,2H),4.75(s,2H),3.87(s,2H),2.50(s,3H).

[0169] Example 11

[0170] Synthesis route:

[0171]

[0172] Step 1: Synthesis of compound WX011-2

[0173] Compound WX011-1 (2 g, 10.75 mmol) was dissolved in dichloromethane (20 mL). The system was cooled to 0°C and diethylaminosulfur trifluoride (2.25 g, 13.98 mmol, 1.85 mL) was added. The reaction mixture was warmed to 20°C and stirred for 16 hours. The reaction mixture was added to a saturated sodium bicarbonate solution (10 mL) and extracted with ethyl acetate (10 mL x 3). The organic phase was concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (0-30% ethyl acetate in petroleum ether) to obtain compound WX011-2, MS m / z (ESI): 209.9 [M+H] + .

[0174] Step 2: Synthesis of compound WX011-3

[0175] Compound WX011-2 (1.5 g, 7.21 mmol), N,N-dimethylethylenediamine (64 mg, 721.14 μmol, 77.62 μL), and cuprous oxide (52 mg, 360.57 μmol) were dissolved in ethylene glycol (15 mL). Aqueous ammonia (9.64 g, 76.98 mmol, 10.59 mL, 28% content) and potassium carbonate (199 mg, 1.44 mmol) were added. The reaction mixture was stirred at 120°C for 12 hours. After cooling to room temperature, the mixture was added to water (20 mL) and extracted with ethyl acetate (20 mL x 3). The organic phases were combined and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (0-30% ethyl acetate in petroleum ether) to obtain compound WX011-3. 1 H NMR (400MHz, CDCl3) δ = 7.57-7.53 (m, 1H), 6.95 (d, J = 7.2Hz, 1H), 6.63-6.29 (m, 2H), 4.68 (s, 2H).

[0176] Step 3: Synthesis of compound WX011-4

[0177] Compound WX011-3 (0.3 g, 2.08 mmol) was dissolved in acetic acid (1.5 mL), and dichloromethane (1.5 mL) was added. The system was cooled to 0°C, and N-iodosuccinimide (468 mg, 2.08 mmol) was added. The resulting reaction solution was stirred for 2 hours. Water (10 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (10 mL x 2). The organic phases were combined and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (0-30% ethyl acetate in petroleum ether) to obtain compound WX011-4. MS m / z (ESI): 270.9 [M+H] + . 1 H NMR (400MHz, CDCl3) δ = 7.52 (d, J = 8.4Hz, 1H), 6.44 (t, J = 54.0Hz, 1H), 6.11 (d, J = 7.6Hz, 1H), 4.51 (s, 2H).

[0178] Step 4: Synthesis of compound WX011

[0179] Compound WX011-4 (120 mg, 444.41 μmol) and compound WX004-2 (119 mg, 533.30 μmol) were dissolved in THF (5 mL). Triethylamine (360 mg, 3.56 mmol), cuprous iodide (13 mg, 66.66 μmol), and 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (32 mg, 66.66 μmol) were added. After nitrogen purge, bis(acetonitrile)palladium dichloride (12 mg, 44.44 μmol) was added. The atmosphere was purged with nitrogen again, and the reaction mixture was stirred at 40°C for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (0-50% ethyl acetate in petroleum ether) to obtain compound WX011. MS m / z (ESI): 365.9 [M+H] + . 1 H NMR (400MHz, CDCl3) δ = 8.20 (d, J = 4.0Hz, 1H), 7.64-7.55 (m,2H),7.50-7.46(m,2H),7.44-7.40(m,2H),7.05-6.78(m,3H),6.56(d,J=8.4Hz,1H),5.40(s,2H),4.81(s,2H),3.88(s,2H).

[0180] Example 12

[0181] Synthesis route:

[0182]

[0183] Step 1: Synthesis of compound WX012-2

[0184] Compound WX012-1 (CAS: 34160-40-2, 2.6 g, 13.98 mmol) was dissolved in methanol (40 mL), and sodium borohydride (529 mg, 13.98 mmol) was added. The reaction solution was stirred at 20°C for 0.5 hours. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL x 2). The organic phases were combined, washed with saturated aqueous sodium chloride (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound WX012-2. 1 H NMR (400MHz, CDCl3) δ = 7.58-7.53 (m, 1H), 7.38 (d, J = 7.6Hz, 1H), 7.31 (d, J = 7.6Hz, 1H), 4.75 (s, 2H).

[0185] Step 2: Synthesis of compound WX012-3

[0186] At -78°C under nitrogen, compound WX012-2 (2.55 g, 13.56 mmol) was dissolved in dichloromethane (40 mL) and slowly added dropwise to a solution of diethylaminosulfur trifluoride (6.56 g, 40.69 mmol, 5.38 mL) in dichloromethane (65 mL). The resulting reaction solution was stirred for 1 hour, then warmed to 20°C and stirred for 15 hours. The reaction solution was poured into ice water (200 mL) with stirring and extracted with dichloromethane (50 mL x 2). The organic phases were combined, washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated by COMBI-FLASH column chromatography (eluent: petroleum ether:ethyl acetate = 100:0 to 2:1) to obtain compound WX012-3. 1 HNMR (400MHz, CDCl3) δ = 7.67-7.61 (m, 1H), 7.46-7.43 (m, 2H), 5.46 (d, J = 46.4Hz, 2H).

[0187] Step 3: Synthesis of compound WX012-4

[0188] Compound WX012-3 (2 g, 10.53 mmol), aqueous ammonia (15.81 g, 126.31 mmol, 17.37 mL, 28% content), and potassium carbonate (291 mg, 2.11 mmol) were added to ethylene glycol (20 mL). Cuprous oxide (75 mg, 526.28 μmol) and dimethylethylenediamine (93 mg, 1.05 mmol, 114.97 μL) were added with stirring. The resulting reaction solution was stirred at 130°C for 12 hours. After cooling to room temperature, water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL x 2). The organic phases were combined, washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by COMBI-FLASH column separation (eluent: petroleum ether:ethyl acetate = 100:0 to 2:1) to obtain compound WX012-4. 1 HNMR (400MHz, CDCl3) δ = 7.41-7.37 (m, 1H), 6.69 (d, J = 7.2Hz, 1H), 6.37 (d, J = 8.0Hz, 1H), 5.22 (d, J = 47.2Hz, 2H), 4.49 (s, 2H).

[0189] Step 4: Synthesis of compound WX012-5

[0190] Compound WX012-4 (210 mg, 1.66 mmol) was dissolved in dichloromethane (1 mL) and glacial acetic acid (1 mL), and N-iodosuccinimide (374 mg, 1.66 mmol) was added. The reaction was stirred at 20°C for 1 hour. Dichloromethane (10 mL) was added to the reaction solution, washed with saturated aqueous sodium sulfite solution (20 mL × 2) and saturated aqueous sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by COMBI-FLASH separation (eluent: petroleum ether: ethyl acetate = 100:0 to 2:1) to obtain compound WX012-5. MS m / z (ESI): 252.8 [M+H] + . 1 H NMR (400MHz, CDCl3) δ=7.68 (d, J=8.8Hz, 1H), 6.21 (d, J=8.4Hz, 1H), 5.32 (d, J=47.2Hz, 2H), 4.68 (s, 2H).

[0191] Step 5: Synthesis of compound WX012

[0192] Compound WX012-5 (100 mg, 396.78 μmol), WX004-2 (133 mg, 595.17 μmol), N,N-diisopropylethylamine (205 mg, 1.59 mmol, 276 μL), cuprous iodide (7 mg, 39.68 μmol), and dichlorobis(triphenylphosphine)palladium (14 mg, 19.84 μmol, 0.05 eq) were added to DMF (2 mL). The atmosphere was purged with nitrogen three times and stirred at 30°C for 2 hours. Ethyl acetate (10 mL) was added to the reaction solution, and the mixture was washed with water (10 mL x 2) and saturated sodium chloride solution (20 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by COMBI-FLASH separation (petroleum ether:ethyl acetate = 100:0 to 2:1) to obtain compound WX012. MS m / z(ESI):347.9[M+H] + . 1 HNMR (400MHz, DMSO-d6)δ=8.19-8.18(m,1H),7.75-7.70(m,1H),7.48-7.39(m,5H),7.01-6.98(m ,1H),6.87(d,J=8.0Hz,1H),6.46-6.45(m,3H),5.34(s,2H),5.32(d,J=47.6Hz,2H),3.90(s,2H).

[0193] Example 13

[0194] Synthesis route:

[0195]

[0196] NaI (3.18 g, 21.25 mmol) was added to a reaction flask, followed by compound WX004 (6.7 g, 21.25 mmol). After nitrogen substitution, anhydrous THF (70 mL) and WX013-1 (16.49 g, 63.74 mmol) were added. The resulting reaction solution was stirred at 25°C for 16 hours. Ethyl acetate (50 mL) was added to the reaction solution, and the mixture was washed with saturated sodium bicarbonate solution (30 mL x 2). The organic phase was concentrated under reduced pressure to obtain crude product WX013-2. Crude product WX013-2 (10 g, 18.60 mmol) was dissolved in DCM (100 mL), the system was cooled to 0°C, and TFA (25 mL, 337.65 mmol) was added. The resulting reaction solution was stirred at 25°C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the crude product. Water (20 mL) was added, the pH was adjusted to 7-8 with aqueous ammonia, and the mixture was washed with ethyl acetate (30 mL x 3). The aqueous phase was purified by column chromatography (reverse-phase C18 column, 40 g, eluent: acetonitrile / water (0.1% ammonia), gradient: 0-20%). The resulting fraction was concentrated under reduced pressure to remove the acetonitrile, and a small amount of formic acid was added to obtain WX013. MS m / z (ESI): 426.1 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ=8.14-8.10(m,2H),7.80-7.76(m,2H),7.46-7.40(m,4H),7.0 9-7.01(m,2H),6.90(d,J=8.4Hz,1H),5.64(d,J=8.0Hz,2H),5.29(s,2H),3.84(s,2H).

[0197] Test Example 1. Minimum Inhibitory Activity Test of Antifungal Drugs

[0198] 1. Purpose of the experiment

[0199] The minimum inhibitory concentration (MIC) and minimum effective concentration (MEC) of the test drugs against fungi were determined.

[0200] 2. Experimental strains and test culture medium

[0201] Experimental strains: Candida parapsilosis ATCC 22019; Candida albicans ATCC MYA-2876; Candida albicans WX-CA009; Candida glabrata ATCC15126; Candida tropicalis ATCC 750;

[0202] Cryptococcus neoformans H99 ATCC 208821;

[0203] Aspergillus fumigatus ATCC-MYA-4609; Aspergillus flavusATCC MYA-1004

[0204] Test medium: RPMI1640 (containing 0.165 M MOPS, pH 7.0)

[0205] 3. Experimental Procedure

[0206] 3.1. Preparation of compound master plate

[0207] On the day of the experiment, dissolve the compound in the vial in 100% DMSO to a stock solution concentration of 6.24 mg / mL. Then dilute 10-fold with DMSO to 0.624 mg / mL for later use.

[0208] In a 96-well microplate (V-bottom), serially dilute the compound solution (0.624 mg / mL) in DMSO by 2-fold to obtain a 100× working solution (wells 1 to 11). The concentrations are 624, 312, 156, 78, 39, 20, 10, 5, 2.5, 1.25, and 0.625 μg / mL. A 100% DMSO positive control is used (well 12). This is the compound master plate.

[0209] 3.2. Preparation of inoculum

[0210] Streak -80°C frozen bacteria Candida parapsilosis ATCC 22019, Candida albicans ATCCMYA-2876, Candida albicans WX-CA009, Candida glabrata ATCC15126, and Candida tropicalis ATCC 750 onto SDA plates and place in a 35±2°C incubator for aerobic culture for 24 hours.

[0211] The -80°C frozen bacteria Cryptococcus neoformans H99 ATCC 208821 were streaked onto SDA plates and placed in a 35±2°C incubator for aerobic culture for 48 h.

[0212] Streak -80°C frozen Aspergillus fumigatus ATCC-MYA-4609 and Aspergillus flavus ATCC MYA-1004 onto SDA plates and culture in an aerobic incubator at 30±2°C for 6 days.

[0213] On the day of the experiment, for strains Candida parapsilosis ATCC 22019, Candida albicans ATCC MYA-2876, Candida albicans WX-CA009, Candida glabrata ATCC15126, Candida tropicalis ATCC 750, and Cryptococcus neoformans H99 ATCC 208821, the plates were removed, colonies on the plates were picked, and suspended in physiological saline. The turbidity of the bacterial suspension was then adjusted to OD600 = 0.2 using a turbidimeter. This bacterial suspension contained ~3.0 × 10 6 CFU / mL. Then dilute the bacterial suspension with the test medium to a concentration of ~3.0×10 3 CFU / mL. This is the inoculum.

[0214] For strains Aspergillus fumigatus ATCC MYA-4609 and Aspergillus flavus ATCC MYA-1004, after removing the plate, add 3 mL of 0.9% saline containing 0.1% Tween 20 to the plate and gently collect the spores. Count the spores using a hemocytometer and adjust the spore suspension to ~5 × 10 6 The spore suspension was then diluted with test medium to a concentration of 0.8 to 1 × 10 5 Spores / mL.

[0215] MIC and MEC Detection

[0216] Transfer 2 μL of 100× working solution from the compound master plate (prepared in 3.1) to a round-bottom 96-well plate (containing 98 μL of assay medium). Then, add 100 μL of the bacterial inoculum (prepared in 3.2) to each well to create an MIC assay plate. The final assay concentrations of the compounds were 6.24, 3.12, 1.56, 0.78, 0.39, 0.20, 0.10, 0.05, 0.025, 0.0125, and 0.006 μg / mL. 1% DMSO was used as a growth control.

[0217] For strains Candida parapsilosis ATCC 22019; Candida albicans ATCC MYA-2876; Candida albicans WX-CA009; Candida glabrata ATCC15126; and Candida tropicalis ATCC 750, all test plates were placed in an incubator at 35±2°C and aerobically cultured for 24 hours.

[0218] For the mold strains Aspergillus fumigatus ATCC-MYA-4609 and Aspergillus flavus ATCC MYA-1004, all test plates were placed at 35±°C under aerobic conditions for 48 h.

[0219] For Cryptococcus neoformans H99 ATCC 208821, all test plates were placed in an incubator at 35±2°C for aerobic culture for 72 h.

[0220] 3.4. Reading MIC and MEC

[0221] After incubation, the MIC (μg / mL) and MEC (μg / mL) of the test compound against yeast and mold were determined by visual observation or microscopic observation of the test plate according to the standards in Table 1 below.

[0222] Table 1. MIC / MEC criteria for test compounds against fungi

[0223]

[0224] 4. Experimental Results

[0225] Table 2. Antibacterial test results

[0226]

[0227]

[0228] Note: The experimental results are the results of 3 independent experiments, unit: μg / mL

[0229] Conclusion: The compounds of the present invention have good antibacterial activity against Candida, Cryptococcus and Aspergillus.

[0230] Test Example 2: Pharmacokinetic Evaluation Experiment in Mice

[0231] Objective: To determine the plasma concentration of a test compound at different times after intraperitoneal injection in female CD-1 mice using LC / MS / MS. This study aims to investigate the pharmacokinetic behavior of the test compound in mice and evaluate its pharmacokinetic characteristics.

[0232] Drug preparation: Weigh an appropriate amount of sample and prepare a clear or suspended solution.

[0233] Dosage regimen: Two healthy female CD-1 mice were purchased from Beijing Weitonglihua Experimental Animal Co., Ltd., fed a normal diet, and administered intraperitoneally.

[0234] Procedure: 2 hours before dosing, animals were orally administered 1-aminobenzotriazole (ABT) (50 mg / kg, 5 mg / mL in saline). Approximately 30 μL of blood was collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after dosing and placed into commercial anticoagulant tubes pre-coated with EDTA-K2. The tubes were centrifuged for 10 minutes to separate plasma and stored at -60°C. The target compound content in the plasma samples was determined by LC / MS / MS. The results are shown in Table 3.

[0235] Table 3. Pharmacokinetic results in mice

[0236]

[0237] Conclusion: The compound exhibited higher exposure in mice when combined with ABT.

[0238] Test Example 3: Pharmacokinetic evaluation experiments in rats, dogs, and monkeys

[0239] Objective: To evaluate the drugability of compounds by measuring their pharmacokinetic properties in different animal species.

[0240] Experimental materials: CD-1 mice, Sprague-Dawley rats, beagle dogs, and cynomolgus monkeys.

[0241] Drug preparation: Weigh an appropriate amount of sample and prepare a clear or suspended solution.

[0242] Experimental Procedure: The pharmacokinetic characteristics of the compound were tested in animals following intravenous and oral administration using a standard protocol. The candidate compound was formulated as a clear solution (for intravenous administration) or a homogeneous suspension (for oral administration) and administered to the animals as a single dose. Whole blood samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours. The supernatant was separated and plasma samples were obtained by centrifugation at 3200 g for 10 minutes. Plasma concentrations were quantified by LC-MS / MS analysis, and pharmacokinetic parameters such as peak concentration, time to peak concentration, clearance, half-life, and area under the concentration-time curve (AUC) were calculated.

[0243] Table 4 Pharmacokinetic parameters of the compounds of the present invention measured in various species

[0244]

[0245] *The concentration of the parent drug was tested in the oral administration experiment

[0246] Experimental results: As shown in Table 4, WX013 was completely decomposed into the parent drug WX004 within 0.5 h in the plasma of rats, dogs and monkeys.

[0247] Conclusion: The pharmacokinetic properties of the compounds of the present invention are good and meet the requirements for drug development.

[0248] Test Example 4: Mouse Candidemia Efficacy Model

[0249] Experimental animals: Female CD-1 mice, 7 weeks old, 27–29 g, n = 5 or 8;

[0250] Microbial pathogen: Candida albicans ATCC MYA-2876;

[0251] Inoculation level and route: 2.0-4.0E+05 CFU / mouse, infection by tail vein injection;

[0252] Treatment: Treatment started 1 hour after infection, with oral administration of ABT first, followed by intraperitoneal injection of the test compound 2 hours later, once a day for a total of 7 days, with a dosing volume of 10 mL / kg.

[0253] Observation indicators: weight changes and mortality of mice in each group within 7 days after infection.

[0254] Conclusion: CD-1 mice injected intravenously with a specific dose of Candida albicans ATCC MYA-2876 exhibited a 100% mortality rate within 7 days, resulting in severe candidemia. In this model, after oral administration of 50 mg / kg ABT, the test compounds WX004, WX006, and WX009 completely protected mice from candidemia at a low dose of 26 mg / kg (n=5) after infection with Candida albicans ATCC MYA-2876. Furthermore, WX004 achieved a 100% survival rate at a dose of 6 mg / kg (n=8).

[0255] Test Example 5: Efficacy Study on the Candida Vaginal Infection Model in Mice

[0256] Experimental animals: Female C3H / NeH mice, 6–8 weeks old, 19–21 g, n = 5–6;

[0257] Microbial pathogen: Candida albicans ATCC MYA-4788;

[0258] Inoculation level and route: 5.0E+05 CFU / mouse, vaginal instillation infection;

[0259] Test substance: WX004: 20mpk

[0260] Treatment: ABT was administered orally 22 hours after infection, and the test compound was injected intraperitoneally 24 hours after infection. This experiment consisted of a vehicle group and a test compound group, with drug administration once daily for 3 days at a volume of 10 mL / kg.

[0261] Observational indicators: 96 hours after infection, vaginal tissue and vaginal wash fluid were collected from each group of mice for CFU counts. Plasma samples were collected 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours, and 24 hours after the last dose.

[0262] Conclusion: A stable vaginal infection model was established in mice after vaginal instillation of a specific dose of Candida albicans ATCC MYA-4788. The bacterial loads in the vaginal tissue and vaginal wash fluid of the vehicle group were 4.8±0.08 lg and 3.7±0.15 lg, respectively. Compared with the vehicle group, the test compound WX004 at a 20 mpk dose significantly reduced the bacterial loads in the vaginal tissue fluid and vaginal wash fluid by 2.7 lg and 2.5 lg, respectively (P<0.001).

Claims

1. A compound represented by formula (P) or a pharmaceutically acceptable salt thereof, in, Ring A is selected from T1 and T2 are selected from CH and N; L1 is selected from -O-, -CH2O- and -OCH2-, wherein -CH2O- and -OCH2- are optionally substituted with 1 or 2 halogens; Each R1 is independently selected from H, F, Cl, Br, I, OH and NH2; Each R2 is independently selected from F, Cl, OH, NH2 and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 R a replace; R3 is selected from Each R4 is independently selected from H, F, Cl, Br and I; Each R a are independently selected from F, Cl, Br and I; m is selected from 1, 2, 3 and 4; n is selected from 1, 2, 3 and 4; z is selected from 1, 2 and 3; The compound represented by formula (P) does not include 2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein L1 is selected from -O-, -CH2O- and -OCH2-.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein T1 is selected from N.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein T2 is selected from CH.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Each R1 is independently selected from H and F.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Structural unit Selected from 7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Each R2 is independently selected from F, Cl, OH, NH2, CH3, CH2CH3 and CH(CH3)2, wherein CH3, CH2CH3 and CH(CH3)2 are optionally replaced by 1, 2 or 3 R a replace.

8. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Each R2 is independently selected from F, Cl, OH, NH2, CH3, CHF2 and CH2F.

9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Structural unit Selected from 10. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Ring A is selected from 11. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Each R4 is independently selected from H and F.

12. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Structural unit Selected from 13. The compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, which is selected from in, L1, T1, T2, R1, R2, R4 and m are as defined in any one of claims 1 to 12.

14. The following compound or a pharmaceutically acceptable salt thereof:

15. Use of the compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof in the preparation of medicaments for treating fungal infections.

16. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is, in, L2 is L1 is selected from -O-, -CH2O- and -OCH2-; T1, T2, R1, R2 and m are as defined in claim 1.

17. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is, in, L2 is L1 is selected from -O-, -CH2O- and -OCH2-; T1, T2 and R1 are as defined in claim 1.

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