A series of piperidine-substituted benzoic acid compounds and their uses

Abstract

Disclosed herein is a series of piperidine-substituted benzoic acid compounds and uses thereof, specifically, compounds represented by formula (I) and pharma- ceutically acceptable salts thereof: [Formula 1] TIFF2024501562000073.tif5360

Description

[Technical field]

[0001] This invention claims priority to the following: CN202011610857.7, the filing date is December 30, 2020; CN202110266011.4, the filing date is March 11, 2021; CN202110751316.4, the filing date is July 2, 2021; CN202111234085.6, application date is October 22, 2021. The present invention relates to a series of piperidine-substituted benzoic acid compounds, in particular to the compounds represented by formula (I) and pharma- ceutically acceptable salts thereof: [Background technology]

[0002] Immune diseases are diseases caused by an imbalance in immune regulation that affects the body's immune response. The complement system is an important component of the immune system and contains a group of proteins that are normally present in an inactive state, namely complement. Complement is activated by substances such as lipopolysaccharides, polysaccharides, peptidoglycans, teichoic acids, and condensed IgA and IgG4 in the alternative activation pathway to mediate immune and inflammatory responses. Here, the activators directly activate C3 to complete the chain reaction of each component from C5 to C9. Complement factor B, also called the C3 activator precursor, can be cleaved into two fragments, Ba and Bb, by complement factor D, and Bb combines with C3b to form the C3 convertase of the alternative pathway to function. Complement factor B acts on the AP pathway, and by inhibiting the activity of factor B, activation of the API pathway can be prevented without interfering with the CP and LP pathways, thus avoiding the increased risk of infection caused by inhibition of the complement system.

[0003] At present, there is no small molecule factor B inhibitor on the market, and Novartis' factor B inhibitor LNP023 is in phase III clinical research and has been used to treat diseases such as PNH, IgAN, C3G, etc. Therefore, there is an urgent need in the art to develop new small molecule factor B inhibitors of the complement system for increased clinical research and validation, and for use in treating various diseases caused by complement abnormalities. Summary of the Invention

[0004] The present invention provides a compound represented by formula (I) or a pharma- ceutically acceptable salt thereof. [ka] however, L is selected from a single bond, NR4 and O; R1 is fluorine, chlorine, C 1-5 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -C 1-3 Alkyl-C 1-3 Alkoxy, -C 1-3 Alkyl-C 3-6 Cycloalkyl and -C 1-3 alkyl-3 to 6 membered heterocycloalkyl The above C 1-5 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -C 1-3 Alkyl-C 1-3 Alkoxy, -C 1-3 Alkyl-C 3-6 Cycloalkyl and -C 1-3 Alkyl-3 to 6-membered heterocycloalkyl each independently optionally comprises 1, 2 or 3 R a is replaced by R2, R3 and R4 are each independently H and C 1-5 alkyl, 1-5 Alkyl is optionally one, two or three R b is replaced by Each R a and R bare each independently selected from H, F, Cl, Br, and I; The condition is that R1 is unsubstituted C 1-5 When selected from alkyl, R2 and R3 are not simultaneously selected from H.

[0005] In some aspects of the invention, the compound, or a pharma- ceutically acceptable salt thereof, is selected from the formula: [ka] however, L, R1, R2 and R3 are as defined herein; Carbon atoms marked with an "*" are chiral carbon atoms and exist in either (R) or (S) single enantiomeric form or enantiomerically enriched form.

[0006] In some aspects of the invention, the compound or a pharma- ceutically acceptable salt thereof is selected from: [ka] wherein L, R1, R2 and R3 are as defined in the present invention.

[0007] In some embodiments of the present invention, R1 is C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -C 1-3 Alkyl-C 1-3 Alkoxy, -C 1-3 Alkyl-C 3-6 Cycloalkyl and -C 1-3 alkyl-3 to 6 membered heterocycloalkyl; 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -C 1-3 Alkyl-C 1-3 Alkoxy, -C 1-3 Alkyl-C 3-6 Cycloalkyl and -C 1-3 Alkyl-3 to 6-membered heterocycloalkyl each independently optionally comprises 1, 2 or 3 R aand other variables are as defined in the present invention.

[0008] In some embodiments of the invention, R1 is selected from the group consisting of fluorine, chlorine, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, TIFF0007641388000004.tif15134, wherein said CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, TIFF0007641388000005.tif15134 may be independently selected to contain 1, 2 or 3 R a and other variables are as defined in the present invention.

[0009] In some embodiments of the invention, R1 is CH3, CF3, CH2CH3, CH2CHF2, CH2CF3, TIFF0007641388000006.tif17123, with other variables as defined in the present invention.

[0010] In some embodiments of the present invention, R1 is TIFF0007641388000007.tif17123, with other variables as defined in the present invention.

[0011] In some embodiments of the present invention, R1 is C 1-5 alkyl, R2 is selected from H and C 1-5 alkyl; R3 is selected from C 1-5 alkyl, and the other variables are as defined herein.

[0012] In some embodiments of the invention, R1 is selected from CH2CH3, R2 is selected from H and CH3, and R3 is selected from CHF2 and CH3, with the other variables being as defined herein.

[0013] In some embodiments of the invention, R2 is selected from H and CH3, and CH3 is optionally selected from 1, 2 or 3 R band other variables are as defined in the present invention.

[0014] In some embodiments of the present invention, R2 is selected from H, CH3, and CHF2, with other variables as defined herein.

[0015] In some aspects of the invention, R3 is selected from H and CH3, with other variables as defined herein.

[0016] In some aspects of the invention, R4 is selected from H and CH3, with other variables as defined herein.

[0017] In some aspects of the invention, L is selected from a single bond and 0, with the other variables being as defined herein.

[0018] In some embodiments of the present invention, the structural unit TIFF0007641388000008.tif2226 is TIFF0007641388000009.tif82164, with other variables as defined in the present invention.

[0019] In some embodiments of the present invention, the structural unit TIFF0007641388000010.tif2325 is TIFF0007641388000011.tif79160, with other variables as defined in the present invention.

[0020] In some embodiments of the present invention, the structural unit TIFF0007641388000012.tif2225 is TIFF0007641388000013.tif205156, with other variables as defined in the present invention.

[0021] The present invention provides a compound represented by formula (I) or a pharma- ceutically acceptable salt thereof. [ka] however, L is selected from a single bond, NR4 and O; R1 is fluorine, chlorine, C 1-5 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -C 1-3 Alkyl-C 1-3 Alkoxy, -C 1-3 Alkyl-C 3-6 Cycloalkyl and -C 1-3 alkyl-3 to 6 membered heterocycloalkyl; 1-5 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -C 1-3 Alkyl-C 1-3 Alkoxy, -C 1-3 Alkyl-C 3-6 Cycloalkyl and -C 1-3 Alkyl-3 to 6-membered heterocycloalkyl each independently optionally comprises 1, 2 or 3 R a is replaced by R2, R3 and R4 are each independently H and C 1-5 alkyl, 1-5 Alkyl is optionally one, two or three R b is replaced by Each R a and R b are each independently selected from H, F, Cl, Br, and I; The condition is that R1 is C 1-5 When selected from alkyl, R2 and R3 are not simultaneously selected from H.

[0022] Some further aspects of the invention are realized by combining any of the variables described above.

[0023] The present invention further provides a compound represented by the formula: or a pharma- ceutically acceptable salt thereof. [ka] .

[0024] In some embodiments of the present invention, the compound or a pharma- ceutically acceptable salt thereof may be selected from the group consisting of are selected. [ka] [ka] [ka] .

[0025] The present invention provides the use of the compound, or a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament relating to complement factor B.

[0026] The present invention further provides the following test method. Method 1. Drug efficacy test for passive Heymann nephritis (PHN) 1.1 Experimental Objective: To investigate the ability of the compounds of the present invention to improve renal function in rat Heymann nephritis induced by Sheep Anti-Rat Fx1A Serum (Goat Anti-Mouse Fx1A Serum), including assessing the reduction of proteinuria levels and the amelioration of renal tissue damage. 1.2 Experimental animals: Male SD rats, 7-10 weeks old, weighing 200-300g

[0027] 1.3 Experimental process: 1.3.1 Modeling: On day D-2 before administration, rat urine was collected. D1 was the first day of the experiment, and the animals in the control group (group 1) were injected once with 5 mL / kg of sheep non-immune serum via the tail vein, while the animals in the model group and administration groups (groups 2 to 6) were injected once with 5 mL / kg of sheep non-immune serum via the tail vein. Inject 5 mL / kg of Sheep Anti-Rat Fx1A Serum once via the tail vein.

[0028] 1.3.2 Administration: The administration method is intragastric administration twice a day with an 8-hour administration interval, and the administration volume is 10mL / kg. On day D1, 1 hour before modeling, the animals in groups 1 and 2 are administered blank solvent 20% PEG400 / 10% Solutol / 70% water, the animals in group 3 are administered LNP023 (60mpk), and the animals in groups 4 to 6 are administered different concentrations of compound 4B (5mpk, 20mpk and 60mpk). 8 hours after the first administration interval, each group is administered again with the same dose and administration volume as the first time. On days D2 to D14, the animals in each group are administered different compounds or solvents for 14 days (including day 1) continuously according to the dose, volume, administration method and frequency on day D1.

[0029] 1.3.3 Sample Collection: (1) Urine collection: On day D-2 before administration, and on days D4, D6, D8, D11, and D14 after administration, rat urine was collected 2 to 4 hours and 4 to 6 hours after the first administration, transferred to EP tubes, and stored in a refrigerator at -80℃ to -60℃, and used to detect rat urinary protein and creatinine. (2) Collection of kidney tissue: All animals were euthanized by CO2 inhalation anesthesia on day D15 after administration, and both kidneys were collected. The left kidney was cut transversely and the right kidney was cut longitudinally. The transversely cut half (left side) and the longitudinally cut half (right side) were fixed in formalin (put in the same EP tube), and the remaining transversely cut half (left side) and longitudinally cut half (right side) were embedded in OCT with the cut surface facing down (put in the same embedding cassette). After embedding, they were stored in a refrigerator at -80°C to -60°C as soon as possible and used for histopathological scoring.

[0030] 1.3.4 Sample Analysis: Urinary protein and creatinine data were read using an analyzer HITACHI LST008 AS(P). Glomerulopathy (characterized by glomerular hypertrophy, thickening of the basement membrane and Bowman's capsule, and podocyte hypertrophy / roundness) was scored by histopathological analysis, tubular degeneration score was scored by histopathological analysis, and analysis was scored for glomerular C3 deposition by IHC or IF staining. Effect of the Invention

[0031] The compounds of the present invention exhibit significant inhibitory activity against the activation of the alternative pathway in human serum, significantly inhibit LPS-stimulated complement activation, reduce urinary protein levels and improve renal function; PK results show that the compounds of the present invention exhibit longer half-life and higher drug exposure, have excellent in vivo pharmacokinetic properties and have excellent drug discovery properties. Definitions and Explanations

[0032] Unless otherwise stated, the following terms and phrases used herein have the following meanings: A particular term or phrase, unless otherwise defined, should be understood to be of its ordinary definition, not of an indefinite or unclear nature. When a trade name appears in this specification, it refers to the corresponding product or its active ingredient.

[0033] As used herein, "pharmacologically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are within the scope of sound medical judgment, suitable for contact with human and animal tissues, without significant toxicity, irritation, allergic response or other problem or complication, and consistent with a reasonable benefit / risk ratio.

[0034] The term "pharmaceutical acceptable salt" refers to a salt of a compound of the present invention, which is prepared with a relatively non-toxic acid or base with a compound having certain substituents discovered in this invention. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting these compounds with a sufficient amount of base in a separate 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 these compounds with a sufficient amount of acid in a separate solution or in a suitable inert solvent. Examples of pharma- ceutically acceptable acid addition salts include inorganic and organic acid salts, as well as salts of amino acids (e.g., arginine, etc.), and organic acids such as glucuronic acid, such as inorganic acids including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, and the like, and organic acids including, for example, 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, methanesulfonic acid, and the like. Certain compounds of the present invention contain both basic and acidic functional groups and can therefore be converted into any suitable base or acid addition salt.

[0035] The pharma- ceutical acceptable salts of the present invention can be synthesized from parent compounds that contain an acidic or basic group by conventional chemical methods. Typically, such salts are prepared by reacting the compounds in their free acid or base form with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or a mixture of both.

[0036] 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 and other mixtures thereof, such as mixtures enriched in enantiomers or non-enantiomers, and all such mixtures are within the scope of the present invention. Other asymmetric carbon atoms may be present in substituents such as alkyl. All such isomers and mixtures thereof are both within the scope of the present invention.

[0037] Unless otherwise stated, the terms "enantiomers" or "optical isomers" are stereoisomers that are mirror images of each other.

[0038] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" refer to the inability to freely rotate about double bonds or single bonds of ring carbon atoms.

[0039] Unless otherwise explained, the term "diastereomer" is a stereoisomer in which the molecules have two or more centers of chirality and the molecules are not mirror-images of each other.

[0040] Unless otherwise specified, "(+)" means dextrorotatory, "(-)" means levorotatory, and "(±)" means racemic.

[0041] Unless otherwise stated, solid wedge connections ( TIFF0007641388000019.tif1216) and dotted wedge bond ( TIFF0007641388000020.tif914) shows the absolute configuration of one stereocenter with a solid rod bond ( TIFF0007641388000021.tif914) and bar-dotted line bond ( TIFF0007641388000022.tif914) shows the relative arrangement of stereocenters, with wavy lines ( TIFF0007641388000023.tif914) with solid wedge connection ( TIFF0007641388000024.tif914) or dotted wedge bond ( TIFF0007641388000025.tif914) or wavy line ( TIFF0007641388000026.tif914) with solid bar join ( TIFF0007641388000027.tif914) and bar-dotted line bond ( TIFF0007641388000028.tif914).

[0042] Unless otherwise stated, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium and can be rapidly interconverted at room temperature. Tautomers can reach chemical equilibrium of tautomers when possible (e.g., in solution). For example, proton tautomers (also called prototropic tautomers) include interconversions via the transfer of a proton, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions by rearrangement of some bond electrons. Among them, a specific example of keto-enol tautomerization is the interconversion between the two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0043] Unless otherwise explained, the terms "enriched in one isomer," "isomer-enriched," "enriched in one enantiomer," or "enantiomer-enriched" mean that the amount of one isomer or enantiomer is less than 100% and that the amount of such isomer or enantiomer is 60% or more, or 70% or more, or 80% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more, or 99.5% or more, or 99.6% or more, or 99.7% or more, or 99.8% or more, or 99.9% or more.

[0044] Unless otherwise stated, 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 90%. If the isomeric content is 10%, the isomeric or enantiomeric excess (ee value) is 80%. The compounds of the present invention may contain unnatural atomic isotopes at one or more atoms that constitute the compounds. For example, tritium ( 3 H), iodine-125( 125 I) or C-14( 14 Compounds can be labeled with radioisotopes such as radioisotopes C). Also, for example, deuterium can be replaced with hydrogen to form deuterated drugs, where the bond formed between deuterium and carbon is stronger than the bond formed between normal hydrogen and carbon, and compared to non-deuterated drugs, deuterated drugs have the advantages of reduced toxic side effects, increased drug stability, improved therapeutic efficacy, and extended biological half-life of the drug. Conversion of the isotopic composition of the compounds of the invention, whether radioactive or not, is within the scope of the invention.

[0045] The terms "optionally" or "optionally" mean that the following event or circumstance may occur, but does not necessarily occur, and that the description includes the case where the described event or circumstance does not occur, if that event or circumstance occurs.

[0046] The term "substituted" refers to the replacement of any one or more hydrogen atoms at a particular atom with a substituent, which may include deuterium and hydrogen variants, provided that the particular valence state is correct and the compound is stable after substitution. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Substitution with oxygen does not occur in aromatic groups. The term "optionally substituted" refers to either substituted or unsubstituted, and unless otherwise defined, the type and number of substituents are any that are chemically stable and feasible.

[0047] When any variable (e.g., R) occurs more than once in a composition or structure of a compound, its definition is independent in each occurrence. Thus, for example, if a group is substituted with 0-2 R, then said group is optionally substituted with up to 2 R, and each occurrence of R is independently optional. Also, combinations of substituents and / or variants thereof are permissible only if such combinations result in stable compounds.

[0048] When the number of linking groups is 0, for example, -(CRR)0- means that the linking group is a single bond.

[0049] When one of the variables is a single bond, the two groups it links are directly linked; for example, when L in ALZ represents a single bond, the structure is actually AZ.

[0050] If the listed linking group does not specify another linking direction, then the linking direction is arbitrary, for example: In TIFF0007641388000029.tif1229, the linking group L is -MW-, where -MW- constitutes ring A and ring B in the same direction as the reading order from left to right. TIFF0007641388000030.tif1235 and construct ring A and ring B in the reverse order of left-to-right reading. Combinations of the above linking groups, substituents and / or variables thereof are permissible only if such combinations result in stable compounds.

[0051] Unless otherwise specified, when a group has one or more bondable sites, any one or more of the sites of the group can be bonded to other groups by chemical bonds. When the bonding mode of the chemical bond is delocalized and there is an H atom at the bondable site, the number of H atoms at the site is reduced to a group with a corresponding valence according to the number of bonded chemical bonds. The chemical bond that bonds the site to another group is represented by a straight solid bond ( TIFF0007641388000032.tif1012), Linear dashed line combination ( TIFF0007641388000033.tif1012), or wavy line ( For example, the straight solid bond in -OCH3 means that the group is bonded to another group via its oxygen atom. The straight dashed bond in TIFF0007641388000035.tif1213 means that both ends of the nitrogen atom in the group are bonded to other groups. The wavy lines in TIFF0007641388000036.tif1523 indicate that the phenyl group is bonded to another group via carbon atoms at positions 1 and 2. TIFF0007641388000037.tif1520 means that any available site on the piperidinyl group can be bonded to another group by one chemical bond, and at least TIFF0007641388000038.tif1885 contains four bond forms, and even though the H atom is drawn as -N-, TIFF0007641388000039.tif1819 contains TIFF0007641388000040.tif1819This bond type includes groups, but when one chemical bond is connected, one H at that site is reduced to the corresponding monovalent piperidine group.

[0052] Unless otherwise defined, the term "C 1-3 The term "alkyl," alone or in combination with other terms, refers to a straight or branched chain saturated hydrocarbon group composed of 1 to 3 carbon atoms, respectively. Said C 1-3 Alkyl has C 1―2 and C. 2―3 Alkyl, which may be monovalent (e.g., methyl), divalent (e.g., methylene) and polyvalent (e.g., methine). 1-3 Examples of alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl).

[0053] Unless otherwise defined, the term "C 1-5 The term "alkyl," alone or in combination with other terms, refers to a saturated hydrocarbon group having 1 to 5 carbon atoms, either linear or branched. 1-5 Alkyl has C 1-4 , C 1-3 , C 1-2 , C 2-5 , C 2-4 and C5 alkyl, which may be monovalent (e.g., methyl), divalent (e.g., methylene) and polyvalent (e.g., methine). 1―5 Illustrative examples of alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl, t-butyl), pentyl (including n-pentyl, isopentyl, and neopentyl), and the like.

[0054] Unless otherwise defined, the term "C 1―3 "Alkoxy", alone or in combination with other terms, refers to an alkyl group containing 1 to 3 carbon atoms, each of which is linked to the remainder of the molecule via an oxygen atom. 1-3 Alkoxy is C 1-2 , C 2-3 , C3 and C2 alkoxy, etc. 1-3 Examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy or isopropoxy), and the like.

[0055] Unless otherwise defined, "C 3-6 "Cycloalkyl" refers, alone or in combination with other terms, to a saturated monocyclic cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, and is defined as any one of the above C 3-6 Cycloalkyl is C 3-5 , C 4-5 Or C 5-6 cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. C 3-6 Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0056] Unless otherwise defined, the term "3- to 6-membered heterocycloalkyl" by itself or in combination with other terms refers to a saturated monocyclic ring group composed of 3 to 6 ring atoms, of which 1, 2, 3, and 4 ring atoms are independently selected from O, S, and N heteroatoms, and the remainder are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). p and p is 1 or 2.) Additionally, with respect to "3- to 6-membered heterocycloalkyl", a heteroatom can occupy the position connecting the heterocycloalkyl to the rest of the molecule. The 3- to 6-membered heteroaryl group includes 4- to 6-membered, 5- to 6-membered, 4-membered, 5-membered and 6-membered heteroaryl groups, and the like. Illustrative examples of 3-6 membered heteroaryl groups include, but are not limited to, heterobutyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, and the like), tetrahydropyran, piperidinyl (including 1-piperidinyl, 2-piperidinyl, and 3-piperidinyl, and the like), piperazinyl (including 1-piperazinyl and 2-piperazinyl, and the like), morpholinyl (including 3-morpholinyl and 4-morpholinyl, and the like), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, or hexahydropyridazinyl.

[0057] Unless otherwise defined, C n-n+m Or C n -C n+m includes any one specific embodiment of n to n+m carbons, for example, C 1-12 are C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 Including any one of the ranges n to n+m, for example, C 1-12 is C 1-3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C3-12 , C 6-9 , C 6-12 , and C 9-12 Similarly, n-membered to n+m-membered rings represent the number of atoms in the ring from n to n+m. For example, 3- to 12-membered rings are It 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 one of the ranges of n to n+m. For example, a 3- to 12-membered ring includes a 3- to 6-membered ring, a 3- to 9-membered ring, a 5- to 6-membered ring, a 5- to 7-membered ring, a 6- to 7-membered ring, a 6- to 8-membered ring, and a 6- to 10-membered ring.

[0058] The compounds of the present invention can be prepared by a variety of synthetic methods familiar to those skilled in the art, including the specific embodiments given below, embodiments in combination with other chemical synthetic methods, and equivalent alternative methods familiar to those skilled in the art, and preferred embodiments include, but are not limited to, the examples of the present invention.

[0059] All solvents used in this invention can be obtained commercially.

[0060] The present invention uses the following abbreviations: aq stands for water; eq stands for equivalent; DCM stands for dichloromethane; PE stands for petroleum ether; DMSO stands for dimethylsulfoxide; EtOAc stands for ethyl acetate; EtOH stands for ethanol; MeOH stands for methanol; DMF stands for N,N-dimethylformamide; CBz stands for benzyloxycarbonyl, an amine protecting group; BOC stands for tert-butoxycarbonyl, an amine protecting group; rt stands for room temperature; RT stands for retention time; O / N stands for overnight;THF stands for tetrahydrofuran;Boc2O stands for di-tert-butyl dicarbonate;TFA stands for trifluoroacetic acid;HCl stands for hydrochloric acid;DIPEA stands for diisopropylethylamine;TEA stands for triethylamine;NBS stands for N-bromosuccinimide;iPrOH stands for 2-propanol;mp stands for melting point;Pd(PPh3)4 stands for tetrakis(triphenylphosphine)palladium;Pd(dppf)Cl2 .CH2Cl2 represents [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride dichloromethane complex; Pd(dppf)Cl2 represents [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II); PPA represents polyphosphoric acid; NMP represents N-methylpyrrolidone; TBSCl represents tert-butyldimethylsilyl chloride; n-BuLi represents n-butyllithium; TBAF represents tetrabutylammonium fluoride; psi represents pounds per square inch; CO2 represents carbon dioxide; DEA represents diethylamine; PEG300 represents polyethylene glycol 300; Cremphor EL represents polyoxyethylene castor oil; and PBS represents phosphate buffer solution.

[0061] The structure of the compound of the present invention can be confirmed by conventional methods known to those skilled in the art, and when the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means of those skilled in the art. For example, single crystal X-ray diffraction (SXRD), cultured single crystal is collected by Bruker D8 venture diffractometer, light source is CuKα radiation, scanning method: φ / ω scanning, after collecting relevant data, the absolute configuration can be confirmed by direct method (Shelxs97) crystal structure analysis. [Brief description of the drawings]

[0062] [Figure 1] Experimental results of an in vivo PD model of LPS-induced complement activation in mice. [Diagram 2] Experimental results of an in vivo pharmacodynamic model of passive Heymann nephritis in rats. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0063] The present invention will be specifically described below by way of examples, but is not intended to be an adverse restriction of the present invention. The present invention has been described in detail in this specification, and specific embodiments thereof have also been disclosed. It is therefore possible for a person skilled in the art to easily modify or change the specific embodiments of the present invention without departing from the spirit and scope of the present invention. It will be apparent that various changes and modifications can be made to the embodiments of the present invention.

[0064] Reference Example 1: Preparation of Intermediates M-2 and M-1 [ka]

[0065] Step 1 At 20° C., 4-dimethylaminopyridine (3.79 g) and di-tert-butyl dicarbonate (8.12 g) were added to a solution of compound MA (5.00 g) in acetonitrile (30.0 mL), the reaction solution was stirred at 20° C. for 1 hour, diluted with ethyl acetate (100.0 mL), washed successively with 1N hydrochloric acid (30.0 mL) and saturated saline (50.0 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound MB. 1 H NMR(400 MHz,CDCl3) δ 7.52 (d,J=4.0Hz,1H),6.87 (d,J=2.4Hz,1H),6.75 (d,J=2.4Hz,1H),6.48 (d,J=4.0Hz,1H),3.85 (s,3H),2.64 (s,3H),1.65 (s,9H).

[0066] Step 2 At 20°C, oxalyl chloride (5.25g) was added to a solution of N-methylformanilide (5.59g) in dichloromethane (50.0mL). The reaction solution was stirred at 20°C for 14 hours under nitrogen gas atmosphere. The reaction solution was transferred to a constant pressure dropping funnel and slowly added dropwise to a solution of compound MB (9.00g) in dichloromethane (50.0mL) at -14°C, and the reaction solution was stirred at -14°C for 3 hours under nitrogen gas atmosphere. Saturated sodium bicarbonate (50.0mL) was added to quench the reaction, washed with saturated saline (50mL x 2), the organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, the residue was washed with ethanol (20.0mL), filtered, and the cake was dried to obtain compound M-2. 1H NMR (400MHz,CDCl3) δ 10.62(s,1H),7.62 (d,J=3.6 Hz,1H),7.47 (d,J=3.6Hz,1H),6.73 (s,1H),3.95 (s,3H),2.68 (s,3H),1.63(s,9H);LC-MS:m / z=290.1[M+H] + .

[0067] Step 3 Sodium borohydride (980.7 mg) was added to a solution of compound M-2 (3.00 g, 10.37 mmol, 1 eq) in methanol (10.0 mL) at 0°C, and the reaction solution was stirred at 0°C for 2 hours. After the reaction was completed, saturated aqueous ammonium chloride solution (200.0 mL) was added to the reaction solution at 0°C. Then, the mixture was diluted with water (100.0 mL), extracted with ethyl acetate (20.0 mL x 2), washed with water (50.0 mL) and saturated saline (50.0 mL), and the organic phase was combined and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 2.2: 1) to obtain compound MC. 1 H NMR(400MHz,CDCl3) δ 7.5 3(d,J=.60Hz,1H),6.74(s,1H),6.61(d,J=3.60Hz,1H),4.89(s,2H),3.90(s,3H),2.63(s,3H),1.62(s,9H).

[0068] Step 4 At 20°C, (chloromethylene)dimethylammonium chloride (1.41g) was added to a solution of compound MC (1.60g) in methanol (1.2mL) and dichloromethane (20.0mL), and the reaction solution was kept warm and stirred for 1 hour. The mixture was cooled to 0°C, quenched by adding saturated sodium bicarbonate solution (20.0mL), extracted with ethyl acetate (30mL x 3), and the organic phases were combined, washed with saturated saline (20.0mL x 2), and dried over anhydrous sodium sulfate. The mixture was filtered and the filtrate was concentrated under reduced pressure to obtain compound M-1.

[0069] Example 1 Preparation of Compounds 1A, 1B, 1C and 1D [ka] [ka]

[0070] Step 1 Compound 1-1 (3.20 g) was dissolved in tetrahydrofuran (30 mL), the reaction solution was placed at -78 ° C, lithium bis (trimethylsilyl) amide (1 M, 10.5 mL) was added under the protection of nitrogen gas, the reaction system was kept warm and stirred for 1 hour, then the temperature was raised to 0 ° C, and compound 1,1,1-trifluoro-N-phenyl-N- ((trifluoromethyl) sulfonyl) methanesulfonamide (5.10 g) was added. The reaction solution was warmed to 30 ° C and stirred for 12 hours. Water (15 mL) was added to quench, and the mixture was extracted with ethyl acetate (40 mL × 3), and the organic phase was washed with saturated saline (40 mL × 1), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1: 0 to 2: 1) to obtain compound 1-2. 1 H NMR (400 MHz, CDCl3) δ 7.56-7.58(m,2H),7.28-7.38(m,6H),7.17-7.23(m,1H),5.89(br s,1H),5.01-5.19(m,3H),4.22(br s,1H),2.92-2.99(m,1H),2.58-2.73(m,1H),2.20-2.38(m,1H);LC-MS:m / z=467.0[M+H] + .

[0071] Step 2 At 20°C, 1,1'-bis(diphenylphosphino)ferrocenedichloropalladium(II) dichloromethane complex (175.1mg) and cesium carbonate (1.05g) were added to a solution of compound 1-2 (0.50g) and cyclobutylboronic acid (139.3mg) in toluene (10mL). The reaction solution was heated to 110°C and stirred under nitrogen gas for 14 hours. The reaction was cooled to room temperature, quenched by adding ice water (20mL), extracted with ethyl acetate (25mL x 3), the organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:10) to obtain compound 1-3. 1 H NMR (400 MHz, CDCl3) δ ppm 7.30-7.67(m,9H),5.37-5.78(m,2H),5.18-5.24(m,1H),5.06-5.17(m,1H),4.17-4.38(m,1H),2.86-3.01( m,2H),2.16-2.28(m,1H),2.08-2.17(m,2H),1.88-2.02(m,4H),1.71-1.82(m,1H);LC-MS:m / z=373.1[M+H] + .

[0072] Step 3 To a mixed solution of compound 1-3 (0.34 g) in isopropanol (2 mL), dioxane (2 mL) and water (4 mL), barium hydroxide (1.16 g) was added, and the reaction solution was heated to 100° C. and stirred for 20 hours. The reaction solution was cooled to room temperature, adjusted to pH<7 with 50% aqueous potassium hydrogen sulfate, extracted with ethyl acetate (40 mL×3), and the organic phases were combined, washed once with water (30 mL) and saturated saline (25 mL), and dried over anhydrous sodium sulfate. Compound 1-4 was obtained by filtration and concentration under reduced pressure. LC-MS: m / z=392.1 [M+H] + .

[0073] Step 4 At 20°C, trimethylsilyldiazomethane (2M, 919μL) was added to a mixed solution of compound 1-4 (0.36g) in methanol (1.2mL) and toluene (3.6mL), and the reaction solution was stirred at 20°C for 1 hour. The reaction solution was cooled to 0°C, quenched by adding acetic acid, diluted with water (40mL), extracted with ethyl acetate (35mL×3), and the organic phases were combined and washed successively with saturated aqueous sodium bicarbonate solution (25mL), water (25mL), and saturated saline (25mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (ethyl acetate:petroleum ether=1:3) to obtain compound 1-5. LC-MS: m / z =406.1[M+H] + .

[0074] Step 5 At 20° C., wet palladium on carbon (10%, 0.20 g) and hydrogen chloride-dioxane solution (4 M, 0.04 mL) were added to a solution of compound 1-5 (0.24 g) in ethanol (4 mL), and the reaction solution was stirred at 20° C. under an atmosphere of hydrogen gas (15 psi) for 1 hour. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 1-6. LC-MS: m / z=274.1 [M+H] + .

[0075] Step 6 Compound M-1 (1.01 g), cesium carbonate (1.64 g) and potassium iodide (334.0 mg) were added to a solution of compound 1-6 (0.55 g) in N,N-dimethylformamide (15 mL) at 20° C. The reaction solution was stirred at 20° C. for 15 hours, poured slowly into water (80 mL), extracted with ethyl acetate (90 mL×3), and the organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:1). The crude product was further separated and purified by high performance liquid chromatography (column: Phenomenex Gemini-NX 80 x 30 mm x 3 μm; mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; gradient: acetonitrile%: 57% to 87%) to obtain compound 1-7, which was further separated chirally (column: DAICEL CHIRALPAK AD (250 mm x 30 mm, 10 μm; mobile phase: A: carbon dioxide, B: [0.1% ammonium water-isopropanol]; gradient: B%: 35% to 35%) to obtain compounds 1-7A, 1-7B, 1-7C, and 1-7D.

[0076] SFC analysis and detection method: Column: Chiralpak AD-3 150mm×4.6mm ID, 3μm, Mobile phase: A: carbon dioxide, B: isopropanol (0.05% diethylamine), Gradient: B%: 5%-40% gradient flow for 5 min, 40% for 5 min, 5% for 2.5 min, Flow rate: 2.5mL / min. Compound 1-7A retention time: 4.238min, ee: 100%; Compound 1-7B retention time: 4.946min, ee: 100%; Compound 1-7C retention time: 5.530min, ee: 99.4%; Compound 1-7D retention time: 5.954min, ee: 100%.

[0077] Two-dimensional NMR analysis confirmed that the carbon-hydrogen bond of the piperidine ring of compound 1-7C bonded to the 4-membered cyclobutyl has a NOE correlation with the carbon-hydrogen bond of the benzoic acid, and that it is a cis structure. The carbon-hydrogen bond of the piperidine ring of compound 1-7D bonded to the 4-membered cyclobutyl has a NOE correlation with the carbon-hydrogen bond of the benzoic acid, and that it is a cis structure. LC-MS: m / z=547.2[M+H] + .

[0078] Step 7 Lithium hydroxide monohydrate (3.5 mg) was added to a mixed solution of compound 1-7A (15.0 mg) in methanol (1 mL) and tetrahydrofuran (0.5 mL), and the reaction solution was stirred at 50°C for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by high performance liquid chromatography (column: Phenomenex Gemini-NX C18 (80 x 30 mm x 3 μm); mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile %: 16% to 86%) to obtain compound 1A. LC-MS: m / z = 433.2 [M + H] + .

[0079] Lithium hydroxide monohydrate (4.6 mg) was added to a mixed solution of compound 1-7B (20.0 mg) in methanol (1 mL) and tetrahydrofuran (0.5 mL), and the reaction solution was stirred at 50° C. for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (column: Phenomenex Gemini-NX C18 (80×30 mm×3 μm); mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonite Compound 1B was obtained by separation and purification using ethyl acetate (16%~86%). LC-MS: m / z=433.3[M+H] + .

[0080] Lithium hydroxide monohydrate (23.0 mg) was added to a mixed solution of compound 1-7C (0.10 g) in methanol (1 mL) and tetrahydrofuran (0.5 mL), and the reaction solution was stirred at 50° C. for 15 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by high performance liquid chromatography (column: Phenomenex Gemini-NX C18 (80×30 mm×3 μm); mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 16% to 86%) to obtain compound 1C. 1 H NMR (400 MHz,DMSO-d6) δ ppm 10.80(s,1H),7.95(d,J=8.80Hz,2H),7.64(d,J=7.60Hz,2H),7.24(t,J=2.80Hz,1H), 6.65(s,1H),6.45(t,J=2.40Hz,1H),3.70(s,3H),3.54(d,J=12.00Hz,1H),3.10-3.30 (m,2H),2.78-2.81(m,1H),2.50-2.55(m,1H),2.41(s,3H),1.56-2.01(m,8H),1.48-1 .51(m,1H),1.34(m,1H),1.02-1.11(m,1H),0.81-0.93(m,1H);LC-MS:m / z=433.3[M+H] + .

[0081] Lithium hydroxide monohydrate (23.0 mg) was added to a mixed solution of compound 1-7D (0.10 g) in methanol (1 mL) and tetrahydrofuran (0.5 mL), and the reaction solution was stirred at 50° C. for 15 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by high performance liquid chromatography (column: Phenomenex Gemini-NX C18 (80×30 mm×3 μm); mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 16% to 86%) to obtain compound 1D. 1H NMR(400 MHz,DMSO-d6) δ ppm 10.80(s,1H),7.95(d,J=8.40Hz,2H),7.64(d,J=7.60Hz,2H),7.24(t,J=2.80Hz,1H ),6.65(s,1H),6.43-6.47(m,1H),3.70(s,3H),3.54(d,J=11.60Hz,1H),3.10-3.30 (m,2H),2.78-2.81(m,1H),2.50-2.55(m,1H),2.41(s,3H),1.54-2.01(m,8H),1.50 (m,1H),1.34(m,1H),1.02-1.11(m,1H),0.81-0.96(m,1H);LC-MS:m / z=433.2[M+H] + .

[0082] Example 2 Preparation of Compounds 2A, 2B, 2C and 2D [ka] [ka]

[0083] Step 1 At 0°C, a dichloromethane solution of titanium tetrachloride (1M, 1.2mL) was dissolved in tetrahydrofuran (1mL), and a tetrahydrofuran (1mL) solution of compound 1-1 (0.20g) and dimethyl maleate (79.0mg) was added, and the reaction solution was stirred at 0°C for 1 hour. Pyridine (236.6mg) was added, and the reaction solution was stirred at 20°C for 15 hours. The reaction solution was poured into 10% citric acid, adjusted to pH<7, and extracted with ethyl acetate (20mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (ethyl acetate:petroleum ether=1:3) to obtain compound 2-2. 1H NMR (400 MHz, CDCl3) δ ppm 7.52(d,J=8.40Hz,2H),7.08-7.32(m,7H),5.23-5.42(m,1H),5.06-5.12(m,1H),4.94-5.05(m,1H),4.01-4.11(m,1H), 3.66-3.72(m,6H),3.29-3.39(m,1H),3.16-3.27(m,1H),2.74-2.90(m,2H),2.54-2.65(m,1H);LC-MS:m / z=449.1[M+H] + .

[0084] Step 2 A solution of compound 2-2 (0.20 g) in methanol (2 mL) was added with sodium borohydride at 0° C. The reaction solution was warmed to 25° C. and stirred for 3 hours. The reaction solution was quenched by adding ice water (30 mL), extracted with dichloromethane (30 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 2-3. LC-MS: m / z=451.4 [M+H] + .

[0085] Step 3 Sodium borohydride (923.7 mg) was added to a solution of compound 2-3 (1.10 g) in ethanol (15 mL) at 25°C. The reaction solution was heated to 70°C and stirred for 2 hours. The reaction solution was cooled to room temperature, quenched by adding ice water (50 mL), extracted with ethyl acetate (50 mL), and the organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:0) to obtain compound 2-4. LC-MS: m / z = 395.1 [M + H] + .

[0086] Step 4 Sodium hydrogen (purity: 60%, 60.8 mg) was added to a solution of compound 2-4 (0.50 g) in tetrahydrofuran (6 mL) at 0°C. The reaction solution was stirred at 0°C for 0.5 hours, methanesulfonyl chloride (0.24 g) was added, the reaction solution was warmed to 25°C, and stirred for 1.5 hours. Ice water (5 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 2:1) to obtain compound 2-5. LC-MS: m / z = 473.1 [M + H] + .

[0087] Step 5 At 0°C, sodium hydrogen (purity: 60%, 330.1 mg) was added to a solution of compound 2-5 (1.30 g) in tetrahydrofuran (40 mL), and the reaction solution was warmed to 20°C and stirred for 1 hour. Next, the reaction solution was heated to 60°C and stirred for 16 hours. The reaction solution was cooled to room temperature, quenched by adding ice water (60 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 2:1) to obtain compound 2-6. LC-MS: m / z = 377.1 [M + H] + .

[0088] Step 6 To a mixture of compound 2-6 (0.55 g) in isopropanol (2 mL), dioxane (2 mL) and water (4 mL), barium hydroxide (1.85 g) was added, and the reaction solution was heated to 100° C. and stirred for 14 hours. The reaction solution was cooled to room temperature, and a 50% aqueous potassium hydrogen sulfate solution was added to adjust the pH to <7, and extracted with ethyl acetate (40 mL×3). The organic phases were combined, washed once with water (40 mL) and saturated saline (45 mL), and dried over anhydrous sodium sulfate. The mixture was filtered and the filtrate was concentrated under reduced pressure to obtain compound 2-7 (0.58 g). LC-MS: m / z=396.1 [M+H] + .

[0089] Step 7 At 20°C, (trimethylsilyl)diazomethane (2M, 1.47mL) was added to a mixed solution of compound 2-7 (0.58g) in methanol (1.5mL) and toluene (4.5mL), and the reaction solution was stirred at 20°C for 2 hours. The mixture was cooled to 0°C, quenched by adding acetic acid, diluted with water (40mL), extracted with ethyl acetate (45mL x 3), and the organic phases were combined and washed successively with saturated aqueous sodium bicarbonate solution (45mL), water (45mL), and saturated saline (50mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 2:1). Compound 2-8 was obtained by separation and purification. LC-MS: m / z=410.1[M+H] + .

[0090] Step 8 At 20° C., wet palladium carbon (content: 10%, 0.2 g) was added to a solution of compound 2-8 (0.40 g) in methanol (5 mL), and the reaction solution was stirred at 20° C. under an atmosphere of hydrogen gas (15 psi) for 1 hour. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 2-9. LC-MS: m / z=276.1 [M+H] + .

[0091] Step 9 Compound M-1 (0.65 g), cesium carbonate (769.2 mg) and potassium iodide (156.8 mg) were added to a solution of compound 2-9 (0.26 g) in N,N-dimethylformamide (6 mL) at 20° C. The reaction solution was stirred at 20° C. for 16 hours. The reaction solution was slowly poured into water (50 mL), extracted with ethyl acetate (20 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was first purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:2), and then separated and purified by high performance liquid chromatography (column: Phenomenex Gemini-NX C18 (80 × 30 mm × 3 μm); mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 57% ~ 87%) to obtain compound 2-10, which was then separated by chiral (column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm); mobile phase: A phase: carbon dioxide, B phase: [0.1% ammonia water-isopropanol]; B%: 40% ~ 40%) to obtain compound 2-10A, compound 2-10B, compound 2-10C, and compound 2-10D.

[0092] SFC analysis and detection method: Column: Chiralpak AD-3 150 mm × 4.6 mm ID, 3 μm, Mobile phase: A: carbon dioxide, B: isopropanol (0.05% diethylamine), Gradient: Mobile phase B: 5% to 40% gradient flow for 5 min, hold at 40% for 5 min, hold at 5% for 2.5 min, Flow rate: 2.5 mL / min. Compound 2-10A retention time: 4.333 min, ee: 100%); Compound 2-10B retention time: 4.835 min, ee: 99.5%; Compound 2-10C retention time: 5.299 min, ee: 99.1%; Compound 2-10D retention time: 5.698 min, ee: 98.1%. LC-MS: m / z = 549.2 [M+H] + .

[0093] Step 10 Lithium hydroxide monohydrate (18.4 mg) was added to a mixed solution of compound 2-10A (80.0 mg) in methanol (3 mL) and tetrahydrofuran (1.5 mL), and the reaction solution was heated to 50°C and stirred for 14 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by high performance liquid chromatography (column: Phenomenex Gemini-NX C18 (80 x 30 mm x 3 μm); mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 15% to 45%) to obtain compound 2A. 1 H NMR (400 MHz,CD3OD) δ ppm 8.12(d,J=8.0Hz,1H),8.12(d,J=8.0Hz,1H),7.61(d,J=8.0Hz,2H),7.31(d,J=2.4Hz,1H),6.77(s,1H),6.33(br s,1H),4.93-4.96(m,2H),4.63(s,1H),4.44-4.80(m,2H),4.22-4.32 (m,1H),4.03-4.09(m,1H),3.77(s,3H),3.67-3.75(m,1H),3.06-3.30 (m,2H),2.51(s,3H),2.42-2.51(m,1H),2.20-2.37(m,1H),1.96-2.09(m,1H),1.72-1.81(m,1H),1.54-1.63(m,1H);LC-MS:m / z=435.2[M+H] + .

[0094] Compound 2-10B (80.0 mg) in methanol (3 mL) and tetrahydrofuran ( Lithium hydroxide monohydrate (18.4 mg) was added to the mixture of 1.5 mL of benzoic acid and 1.5 mL of ethyl acetate, and the reaction solution was stirred at 50°C for 14 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by high performance liquid chromatography (column: Phenomenex Gemini-NX C18 (80 x 30 mm x 3 μm); mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 15%-45%) to obtain compound 2B. As measured by two-dimensional NMR, the carbon-hydrogen bond between piperidine and the four-membered epoxide was an equatorial bond, and the carbon-hydrogen bond between benzoic acid and the piperidine was an axial bond, and compound 2B was in a trans configuration.1 H NMR (400 MHz, CD3OD) δ ppm 8.13 (d,J=8.4Hz,2H), 7.61 (d,J=8.4 Hz,2H),7.30(d,J=3.2Hz,1H),6.72(s,1H),6.31(d,J=2.8Hz,1H),4.85-4.91(m,2H ),4.36-4.46(m,3H),4.27(d,J=12.8Hz,1H),4.03(d,J=12.8Hz,1H),3.74(s,3H),3. 66-3.72(m,1H),3.16-3.25(m,2H),2.50(s,3H),2.42-2.47(m,1H),2.27-2.30(m,1 H),2.03-2.10(m,1H),1.78-1.81(m,1H),1.59-1.63(m,1H);LC-MS:m / z=435.2[M+H] + .

[0095] Lithium hydroxide monohydrate (4.6 mg) was added to a mixed solution of compound 2-10C (0.02 g) in methanol (1 mL) and tetrahydrofuran (0.5 mL), and the reaction solution was stirred at 50 ° C for 14 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by high performance liquid chromatography (column: Phenomenex Gemini-NX 80 × 30 mm × 3 μm; mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile %: 15% to 45%) to obtain compound 2C. As measured by two-dimensional NMR, the carbon-hydrogen bond between piperidine and the four-membered epoxide and the carbon-hydrogen bond between benzoic acid were both axial bonds, and compound 2C was in a cis configuration. 1H NMR (400 MHz, CD3OD) δ ppm 8.13(d,J=8.0Hz,2H),7.61(d,J=8.0Hz,2H),7.31(d,J=3.2Hz,1H),6.75(s,1H),6.32(s,1H) ,4.71-4.81(m,2H),4.47-4.56(m,2H),4.29-4.39(m,2H),3.93-3.958(m,1H),3.42-3.48(m,1 H),3.75(s,3H),3.11-3.21(m,1H),2.79-2.83(m,1H),2.51(s,3H),2.16-2.29(m,1H),1.91- 2.01(m,1H),1.79-1.85(m,1H),1.59-1.74(m,1H),1.39-1.46(m,1H);LC-MS:m / z=435.2[M+H] + .

[0096] Lithium hydroxide monohydrate (4.6 mg) was added to a mixed solution of compound 2-10D (0.02 g) in methanol (1 mL) and tetrahydrofuran (0.5 mL), and the reaction solution was stirred at 50°C for 14 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by high-performance liquid chromatography (column: Phenomenex Gemini-NX C18 (80 x 30 mm x 3 μm); mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 15% to 45%) to obtain compound 2D. Measurement by two-dimensional NMR revealed that the carbon-hydrogen bond between piperidine and the four-membered epoxide and the carbon-hydrogen bond with benzoic acid were both axial bonds, and 2D was in a cis configuration. 1H NMR(400 MHz,CD3OD) δ ppm 8.14(d,J=8.0Hz,2H),7.62(d,J=8.0Hz,2H),7.31(d,J=3.2Hz,1H),6.75(s,1H ),6.32(s,1H),4.71-4.81(m,2H),4.46-4.57(m,2H),4.26-4.45(m,2H),3.98- 4.06(m,1H),3.76(s,3H),3.41-3.49(m,1H),3.14-3.26(m,1H),2.76-2.88(m, 1H),2.51(s,3H),2.16-2.29(m,1H),1.95-2.05(m,1H),1.78-1.87(m,1H),1.56 -1.75(m,1H),1.36-1.51(m,1H);LC-MS:m / z=435.2[M+H] + .

[0097] Example 3 Preparation of Compounds 3A, 3B, 3C and 3D [ka] [ka]

[0098] Step 1 Compound 1-2 (66.0 mg) and cyclopropylboronic acid (24.3 mg) were dissolved in 1,4-dioxane (1 mL), potassium phosphate (96.1 mg) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (5.8 mg) were added, and the reaction solution was stirred at 90°C under nitrogen gas protection for 4.5 hours. The reaction solution was concentrated under reduced pressure, quenched by adding water (5 mL), extracted with ethyl acetate (10 mL), and the organic phase was washed with saturated saline, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel thin layer chromatography (petroleum ether:ethyl acetate). =2:1) ​​to obtain compound 3-3. 1H NMR(400 MHz,CDCl3) δ 7.38-7.61(m,2H),7.08-7.33(m,7H),5.44(br s,1H),4.94-5.18(m,2H),3.97-4.24(m,1H),2.75-2.96(m,1H),2.05-2.23(m,1H),1.77-1.80(m,1H) ,1.31-1.44(m,1H),1.11-1.28(m,1H),0.53-0.68(m,2H),0.37-0.52(m,2H);LC-MS:m / z=359.2[M+H] + .

[0099] Step 2 Compound 3-3 (0.3 g) was dissolved in a mixture of isopropanol, 1,4-dioxane and water (2 mL: 2 mL: 5 mL), and barium hydroxide (573.6 mg) was added to the reaction solution, which was then heated to 100°C and stirred for 12 hours while maintaining the temperature. The reaction solution was cooled to room temperature, and a 50% aqueous potassium hydrogen sulfate solution was added to adjust the pH to 5-6, and dichloromethane (30 mL x 3) was added for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 3-4. LC-MS: m / z = 378.0 [M + H] + .

[0100] Step 3 Compound 3-4 (50.0 mg) was dissolved in a mixture of methanol (1.5 mL) and toluene (0.5 mL), and (trimethylsilyl)diazomethane (2 M, 199 μL) was added at room temperature. The reaction solution was kept warm and stirred for 12 hours. Acetic acid (1 mL) and water (10 mL) were added to quench the reaction, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phase was washed with saturated saline (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified on a silica gel thin-layer chromatography plate (petroleum ether: ethyl acetate = 3:1) to obtain compound 3-5. 1H NMR(400 MHz, CDCl3) δ 7.95-7.97(m,2H),7.28-7.58(m,7H),5.55(br s,1H),5.03-5.24(m,2H),4.09-4.21(m,1H),3.92(s,3H),2.84-3.05(m,1H),2.11-2.19(m,1H),1.84-1.88(m,1H),1.63(br s,1H),1.37-1.53(m,1H),0.38-0.7(m,4H);LC-MS:m / z=392.2[M+H] + .

[0101] Step 4 Compound 3-5 (250.0 mg) was dissolved in a mixture of methanol (3 mL) and ethyl acetate (3 mL), and barium hydroxide (0.10 g) was added. The reaction solution was stirred at 15°C and H2 (15 Psi) for 2 hours. The reaction solution was directly filtered and concentrated under reduced pressure, and the residue was separated and purified by silica gel thin layer chromatography (petroleum ether:ethyl acetate = 1:1) to obtain compound 3-6. LC-MS: m / z = 259.9 [M + H] + .

[0102] Step 5 Compound 3-6 (0.20 g) was dissolved in 1,2-dichloroethane (5 mL), compound M-2 (133.9 mg) and sodium triacetylcyanoborohydride (269.7 mg) were added, and the reaction solution was stirred at 15 ° C for 20 hours. Water (3 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phase was washed with saturated saline (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified by high-performance liquid chromatography (column: Boston Green ODS 150 × 30 mm × 5 μm; mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; B (acetonitrile) %: 43% ~ 73%, 9 min) to obtain compound 3-7. Compound 3-7 was separated and purified by SFC chiral column chromatography (column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm); mobile phase: phase A: carbon dioxide, phase B: [0.1% aqueous ammonia-ethanol]; B%: 35% to 35%) to give compound 3-7A. As a result, compounds 3-7B, 3-7C, and 3-7D were obtained. SFC detection method: Column: Chiralpak AD-3 150 mm x 4.6 mm ID, 3 μm, Mobile phase: A: carbon dioxide, B: isopropanol (0.05% diethylamine), Gradient: B%: 5% to 40% gradient flow for 5 min, hold at 40% for 5 min, hold at 5% for 2.5 min, Flow rate: 2.5 mL / min. Compound 3-7A retention time: 3.781 min, ee: 100%; Compound 3-7B retention time: 4.455 min, ee: 74.9%; Compound 3-7C retention time: 5.030 min, ee: 100%; Compound 3-7D retention time: 5.527 min, ee: 97.2%.

[0103] Step 6 At 20°C, compound 3-7A (10.0 mg) was dissolved in a mixture of tetrahydrofuran (0.2 mL) and methanol (0.4 mL), then lithium hydroxide aqueous solution (1 M, 0.2 mL) was added, and the reaction system was stirred at 20°C for 36 hours. 50% potassium hydrogen sulfate aqueous solution was added to adjust the pH to 6-7, and extracted with ethyl acetate (10 mL). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was separated and purified by high performance liquid chromatography (column: Boston Green ODS 150 x 30 mm x 5 μm; mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; acetonitrile%: 20%-60%) to obtain compound 3A. 1 H NMR(400 MHz,CD3OD) δ 8.24(d,J=8.2Hz,2H), 7.76 (d,J=8.2Hz,2H),7.35(d,J=3.2Hz,1H),6.78(s,1H),6.39(d,J=3.0Hz,1H),4. 79-4.90(m,1H),4.17-4.48(m,2H),3.77(s,3H),3.42-3.70(m,2H),2.52(s,3H) ,2.21-2.39(m,1H),2.02-2.19(m,2H),1.91-1.96(m,1H),1.33-1.55(m,1H),1 .03-1.12(m,1H),0.59-0.69(m,2H),0.09-0.19(m,2H);LC-MS:m / z=419.2[M+H]+ .

[0104] At 20°C, compound 3-7B (20.0 mg) was dissolved in a mixture of tetrahydrofuran (0.3 mL) and methanol (0.6 mL), then lithium hydroxide monohydrate aqueous solution (1 M, 0.3 mL) was added, and the reaction system was stirred at 20°C for 36 hours. A 50% aqueous potassium hydrogen sulfate solution was added to adjust the pH to 6-7, the mixture was concentrated under reduced pressure, and the residue was separated and purified by high performance liquid chromatography (column: Boston Green ODS 150 x 30 mm x 5 μm; mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; acetonitrile%: 20%-60%) to obtain compound 3B. 1 H NMR (400 MHz, CD3OD) δ 8.24(d,J=8.2Hz,2H),7.76(d,J=8.2Hz,2H),7.35(d,J=3.0Hz,1H),6.78(s,1H),6.39(d,J=3.2Hz,1H) ,4.79-4.90(m,1H),4.19-4.44(m,2H),3.77(s,3H),3.44-3.71(m,2H),2.52(s,3H),2.27-2.39(m,1H), 2.03-2.17(m,2H),1.88-1.93(m,1H),1.42-1.53(m,1H),1.07-1.12(m,1H),0.62-0.73(m,2H),0.17-0.26(m,2H);LC-MS:m / z=419.2[M+H] + .

[0105] Compound 3-7C (18.0 mg) was dissolved in a mixture of tetrahydrofuran (0.3 mL) and methanol (0.6 mL) at 20°C, lithium hydroxide monohydrate (1 M, 0.3 mL) was then added, and the reaction system was stirred at 20°C for 36 hours. A 50% aqueous solution of potassium hydrogen sulfate was added to adjust the pH to 6-7, the mixture was concentrated under reduced pressure, and the residue was separated and purified by high performance liquid chromatography (column: Boston Green ODS 150 x 30 mm x 5 μm; mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; acetonitrile%: 20%-60%) to obtain compound 3C. 1 H NMR (400 M Hz,CD3OD) δ 8.23(d,J=8.0Hz,2H),7.67-7.80(m,2H), 7.33 (d,J=3.0Hz,1H),6.77 (s, 1H),6.34(d,J=3.0Hz,1H),4.45-4.55(m,1H),4.34(d,J=12.4Hz,1H),4.13(d, J=12.4Hz,1H),3.76(s,3H),3.53-3.66(m,1H),3.21-3.34(m,1H),2.51(s,3H) ,2.28-2.39(m,1H),1.88-2.04(m,2H),1.63-1.77(m,1H),1.04-1.21(m,1H),0 .55-0.67(m,1H),0.36-0.50(m,2H),0.07-0.28(m,2H);LC-MS:m / z=419.2[M+H] + .

[0106] Compound 3-7D (20.0 mg) was dissolved in a mixture of tetrahydrofuran (0.3 mL) and methanol (0.6 mL) at 20°C, lithium hydroxide monohydrate (1 M, 0.3 mL) was then added, and the reaction system was stirred at 20°C for 36 hours. A 50% aqueous solution of potassium hydrogen sulfate was added to adjust the pH to 6-7, the mixture was concentrated under reduced pressure, and the residue was separated and purified by high performance liquid chromatography (column: Boston Green ODS 150 x 30 mm x 5 μm; mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; acetonitrile%: 20%-60%) to obtain compound 3D. 1 H NMR (400 MHz,CD3OD) δ 8.24(br s,2H),7.75(br s,2H),7.33(br s,1H),6.76(br s,1H),6.34(br s,1H),4.24-4.58(m,2H),4.09-4.21(m,1H),3.76(br s,3H),3.50-3.66(m,1H),2.59-2.65(m,1H),2.50(br s,3H),2.09-2.30(m,1H),1.86-2.07(m,2H),1.59-1.79(m,1H),1.02-1. 30(m,1H),0.35-0.77(m,3H),0.12-0.51(m,2H);LC-MS:m / z=419.2[M+H] + .

[0107] Example 4 Preparation of Compounds 4A and 4B [ka] [ka]

[0108] Step 1 At 25°C, under the protection of nitrogen gas, potassium tert-butoxide (1.40g) was added to a solution of trimethylsulfoxonium iodide (2.50g) in 1,2-dichloroethane (15.0mL), the reaction solution was kept warm and stirred for 0.5 hours, the temperature was cooled to 0°C, a solution of compound 1-1 (3.50g) in 1,2-dichloroethane (15.0mL) was added, the reaction system was warmed to 25°C, and the reaction was stirred for 12 hours. Water (10.0mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (50.0mL x 3). The organic phase was washed with saturated saline, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate, petroleum ether ratio: 100% to 35%) to obtain compound 4-2. LC-MS: m / z = 349.1 [M + H] + .

[0109] Step 2 Compound 4-2 (1.70 g) was dissolved in dichloromethane (20.0 mL), then boron trifluoride diethyl etherate (1.04 g) was added, and the reaction system was stirred at 25 °C for 12 hours. Water (5.0 mL) was added to quench the reaction, followed by extraction with dichloromethane (20.0 mL x 3). The organic phase was washed with saturated saline (20.0 mL x 1), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:0 to 2:1) to obtain compound 4-3. LC-MS: m / z = 349.1 [M + H] + .

[0110] Step 3 At 25°C, under the protection of nitrogen gas, tert-butoxide (744.8mg) was added to a solution of methyltriphenylphosphonium iodide (3.13g) in tetrahydrofuran (15.0mL), the reaction solution was stirred at 25°C for 1 hour, then a solution of compound 4-3 (1.80g) in tetrahydrofuran (10.0mL) was added, and the reaction solution was stirred for 12 hours. Water (10.0mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (20mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 to 3:1) to obtain compound 4-4. 1 H NMR (400 MHz, CDCl3) δ 7.57-7.71(m,2H),7.29-7.47(m,7H),5.73-5.79(m,1H),5.63-5.70(m,1H),5.20(br s,2H),4.96-5.06(m,2H),4.14-4.35(m,1H),2.76-2.90(m,1H),2.36-2.39(m,1H),2.04-2. 18(m,1H),1.79-1.83(m,1H),1.64-1.72(m,1H),1.25-1.46(m,1H);LC-MS:m / z=347.1[M+H] + .

[0111] Step 4 At 20℃, under the protection of nitrogen gas, compound 4-4 (1.20g), zinc copper reagent (4.47g) and phosphorus oxychloride (584.3mg) were dissolved in ethylene glycol dimethyl ether (50.0mL), trichloroacetyl chloride (3.15g) was added, and the reaction solution was stirred at 20℃ for 12 hours. It was concentrated under reduced pressure, and the residue was separated and purified by flash silica gel column chromatography (ethyl acetate / petroleum ether, ethyl acetate ratio: 0-50%) to obtain compound 4-5. LC-MS: m / z=457,459[M+H] + .

[0112] Step 5 Compound 4-5 (1.50 g) was dissolved in dioxane (2.0 mL) at 20°C, and glacial acetic acid (15.75 g) and zinc powder (2.14 g) were added. After the addition was completed, the temperature was raised to 80°C and stirred for 1 hour while maintaining the temperature. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by flash silica gel column chromatography (ethyl acetate / petroleum ether, ethyl acetate ratio: 0-50%) to obtain compound 4-6. 1 H NMR(400 MHz,CDCl3) δ:7.63(d,J=8.4Hz,2H),7.21-7.48(m,7H),5.64(br s,1H),5.19(br s,2H),4.28(br s,1H),3.03-3.20(m,2H),2.64-2.87(m,3H),2.30-2.43(m,1H),2.02-2.19(m,1H),1.67(m,2H),1.21-1.41(m,2H).

[0113] Step 6 At 15°C, compound 4-6 (1.2g) was dissolved in dichloromethane (30.0mL), cooled to -15°C, diethylaminosulfur trifluoride (2.49g) was added, the temperature was slowly raised to 15°C, and stirred for 12 hours. The reaction solution was slowly poured into ice water (100.0mL) to quench, the pH was adjusted to 7 with saturated aqueous sodium bicarbonate, extracted with dichloromethane (100mL x 3), and the organic phases were combined and concentrated under reduced pressure. The residue was separated and purified by flash silica gel column chromatography (ethyl acetate / petroleum ether, ethyl acetate ratio: 0-50%) to obtain compound 4-7. 1 H NMR (400 MHz, CDCl3) δ 7.64(d,J=8.4Hz,2H),7.10-7.50(m,7H),5.62(br s,1H),5.19(br s,2H),4.24(br s,1H),2.74-2.77(m,1H),2.52-2.72(m,2H),2.20-2.35(m,1H),2.06-2.22(m,2H),1.78- 1.91(m,1H),1.59-1.68(m,1H),1.50-1.57(m,1H),1.23-1.39(m,1H),1.07-1.21(m,1H).

[0114] Step 7 At 20°C, compound 4-7 (1.00 g) was dissolved in a mixture of isopropanol (5.0 mL), dioxane (5.0 mL) and water (10.0 mL), and barium hydroxide (960.1 mg) was added. The temperature was raised to 100°C and the mixture was stirred for 12 hours while maintaining the temperature. The reaction solution was cooled to room temperature, poured into water (100.0 mL) to quench the reaction, adjusted to pH 5-6 with 1M hydrochloric acid, extracted with ethyl acetate (100 mL x 3), and the organic phases were combined and concentrated under reduced pressure to obtain compound 4-8. LC-MS: m / z = 430 [M + H] + .

[0115] Step 8 Compound 4-8 (0.98 g) was dissolved in a mixture of methanol (5.0 mL) and toluene (15.0 mL) at 20° C., then trimethylsilyldiazomethane (2 M, 2.45 mL) was slowly added, and the reaction was stirred at 20° C. for 0.5 h. Acetic acid (2.0 mL) was slowly added dropwise to quench the reaction, and then the reaction was concentrated under reduced pressure. The concentrate was subjected to flash silencing. Compounds 4-9 were obtained by separation and purification using silica gel column chromatography (ethyl acetate / petroleum ether, ethyl acetate ratio: 0-50%). The results of two-dimensional NMR spectrum (HSQC, COSY, NOE) analysis showed that the CH bond between piperidine and difluorocyclobutane is an axial bond, and there is NOE correlation with the two hydrogens at the α-position of the benzene ring, and the relative configuration of difluorocyclobutane and methyl benzoate can be determined to be a trans configuration. 1 H NMR(400 MHz,CDCl3) δ 8.04(br d,J=8.4Hz,2H),7.10-7.50(m,7H),5.64(br s,1H),5.21(s, 2H),4.27(br s,1H),3.94(s,3H),2.77-2.92(m,1H),2.51-2.76(m,2H),2.27-2.41(m,1H),2.08-2.26(m,2H), 1.73-1.92(m,1H),1.47-1.69(m,2H),1.30-1.45(m,1H),1.08-1.23(m,1H);LC-MS:m / z=444[M+H] +.

[0116] Step 9 At 25°C, compound 4-9 (900.0 mg) was dissolved in ethyl acetate (15.0 mL), then a solution of palladium hydroxide (180.0 mg) in methanol (5.0 mL) was added, and the reaction system was replaced with hydrogen gas three times, and then the hydrogen gas pressure was maintained at 15 psi, and the mixture was stirred at room temperature for 2 hours. After filtration, the filtrate was concentrated under reduced pressure to obtain compound 4-10. 1 H NMR (400 MHz,CDCl3) δ 8.00(d,J=8.4Hz,2H),7.45(d,J=8.4Hz,2H),3.92-3.98(m,1H),3.91(s,3H),2.953-2.96(m,2H),2.64-2.76(m,2H),2.38-2.5 2(m,1H),2.05-2.25(m,2H),1.80-1.90(m,1H),1.70-1.79(m,2H),1.61-1.68(m,1H),1.43-1.52(m,1H);LC-MS:m / z=310[M+H] + .

[0117] Step 10 At 25°C, compound 4-10 (70.0 mg) and compound M-1 (180.0 mg) were dissolved in N,N-dimethylformamide (3.0 mL), then cesium carbonate (221.2 mg) and potassium iodide (37.6 mg, 226.28 μmol, 1 eq.) were added, and the reaction solution was kept warm and stirred for 12 hours. The reaction solution was poured into water (100 mL) to quench, extracted with ethyl acetate (70 mL x 3), and the organic phases were combined and concentrated under reduced pressure. The residue was separated and purified by silica gel thin layer chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound 4-11, which was further separated chirally (chiral column: Phenomenex-Cellulose-2 (250 mm × 30 mm, 10 μm)); mobile phase: [A: carbon dioxide, B: ethanol (0.1% ammonia water)]; gradient B%: 20% ~ 20%) to obtain compounds 4-11A and 4-11B. SFC analysis and detection method: Column: Cellulose 2 150 mm x 4.6 mm ID, 5 μm, Mobile phase: A: carbon dioxide, B: ethanol (0.05% diethylamine), Gradient: B%: 5%-40% gradient flow for 5 min, 40% for 5 min, 5% for 2.5 min, Flow rate: 2.5 mL / min. Retention time of compound 4-11A: 3.267 min, retention time of compound 4-11B: 3.514 min. LC-MS: m / z=583 [M+H] + .

[0118] Step 11 Compound 4A: At 20°C, compound 4-11A (50.0 mg) was dissolved in a mixture of tetrahydrofuran (1.0 mL), methanol (2.0 mL) and water (1.0 mL), then lithium hydroxide (40.0 mg) was added, the reaction solution was heated to 50°C and stirred for 12 hours. The reaction solution was poured into water (20.0 mL), the pH was adjusted to 4-5 with 4M aqueous acetic acid, extracted with ethyl acetate (30 mL x 3), the organic phase was collected and washed with saturated saline (20.0 mL), It was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was dissolved in a mixed solvent of water (10.0 mL) and acetonitrile (2.0 mL) and lyophilized to obtain compound 4A (SFC detection method: column: Chiralpak AD-3 150 mm × 4.6 mm ID, 3 μm, mobile phase: A: carbon dioxide, B: isopropanol (0.05% diethylamine), gradient: B%: 5% to 40% gradient flow for 5.5 min, hold at 40% for 3 min, hold at 5% for 1.5 min, flow rate: 2.5 mL / min, retention time: 6.521 min, ee: 99.7%). 1H NMR (400MHz, DMSO-d6)δ:10.80 (br s,1H),7.89(d,J=8.4Hz,2H),7.52(br d,J=8.0Hz,2H),7.24(t,J=2.8Hz,1H),6.64(s,1H),6.48(t,J=2.4Hz,1H),3.70(s,3H),3.50-3.60(m,2H),2.67-2.77(m,2H),2 .41(s,3H),2.08-2.25(m,4H),1.71-1.80(m,1H),1.58-1.68(m,1H),1.48-1.58(m,3H),1.34-1.37(m,2H);LC-MS:m / z=469[M+H] + .

[0119] Compound 4B: At 20° C., compound 4-11B (46.0 mg) was dissolved in a mixture of tetrahydrofuran (1.0 mL), methanol (2.0 mL) and water (1.0 mL), and then lithium hydroxide (40.0 mg) was added thereto. The reaction solution was heated to 50° C. and stirred for 12 hours. The reaction solution was poured into water (20.0 mL), the pH was adjusted to 4-5 with 4 M aqueous acetic acid, and extracted with ethyl acetate (30 mL x 3). The organic phase was collected, washed with saturated saline (20.0 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was dissolved in a mixed solvent of water (10.0 mL) and acetonitrile (2.0 mL), and lyophilized to obtain compound 4B (SFC detection method: column: Chiralpak AD-3 150 mm x 4.6 mm ID, 3 μm, mobile phase: A: carbon dioxide, B: isopropanol (0.05% diethylamine), gradient: B%: 5%-40% gradient flow for 5.5 min, hold at 40% for 3 min, hold at 5% for 1.5 min, flow rate: 2.5 mL / min, retention time: 4.832 min, ee: 96.0%). 1H NMR(400 MHz,DMSO-d6) δ:10.80(br s,1H),7.93(d,J=8.0Hz,2H),7.61(br d,J=7.4Hz,2H),7.24(t,J=2.8Hz,1H),6.64(s,1H),6.48(t,J=2.0Hz,1H),3.70(s,3H),3.45 -3.65(m,2H),2.67-2.78(m,2H),2.41(s,3H),2.11-2.28(m,4H),1.71-1.80(m,1H),1.62(br s,1H),1.45-1.58(m,3H),1.30-1.40(m,2H);LC-MS:m / z=469[M+H] + .

[0120] Example 5: Preparation of Compound 4C [ka]

[0121] Step 1 At 25°C, compound 5-1 (1.00 kg) and compound 5-2 (919.93 g) were dissolved in 2-methyltetrahydrofuran (10.0 L), and tetraethoxytitanium (4.81 kg) was added in batches. The reaction solution was heated to 80°C under the protection of nitrogen gas and stirred for 16 hours. The reaction solution was poured into ice water (15.0 L), filtered, the cake was washed with 2-methyltetrahydrofuran (2.0 L x 3), the filtrate was collected, washed with saturated saline (3.0 L), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove 2-methyltetrahydrofuran (8.0 L), and the remaining solution was poured into petroleum ether (10.0 L) at 0°C, stirred for 1 hour, filtered, the cake was washed with petroleum ether (1.0 L x 3), and further dried under reduced pressure to obtain compound 5-3. 1 H NMR (400 MHz, CDCl3) δ:7.96-7.98(m,2H),7.73-7.75(m,2H),2.81(s,3H),1.34(s,9H).

[0122] Step 2 At -20°C, under the protection of nitrogen gas, lithium diisopropylamide (2M tetrahydrofuran solution, 12.08mL) was added dropwise to a solution of compound 5-3 (5.00g) in 2-methyltetrahydrofuran (100.0mL), and after completion of the addition, the reaction solution was stirred at -20°C to -10°C for 0.5 hours, and a solution of compound 5-4 (5.74g) in 2-methyltetrahydrofuran (25.0mL) was added, the temperature was slowly raised, and the mixture was stirred at -20°C to -10°C for 7 hours. The reaction was quenched by adding a saturated aqueous ammonium chloride solution (30.00mL), diluted with water (200.0mL), and extracted with ethyl acetate (100.0mL x 2). The organic phases were combined, washed with saturated brine (100mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, ethyl acetate ratio: 0-20%) to obtain compound 5-5. LC-MS: m / z=439.1

[0123] Step 3 Compound 5-5 (5.00 g) was dissolved in tetrahydrofuran (50.0 mL) at 0°C, sodium borohydride (0.62 g) was added, and the reaction system was stirred at 0°C for 3 hours. Water (30.0 mL) was added to quench the reaction, followed by extraction with ethyl acetate (30.0 mL x 2). The organic phase was washed with saturated saline (30.0 mL x 1), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, ethyl acetate ratio: 0-100%) to obtain compound 5-6. LC-MS: m / z = 399.2 [M + H] + .

[0124] Step 4 At 25°C, under the protection of nitrogen gas, triphenylphosphine (1.18g) and diisopropyl azodicarboxylate (913.3mg) were added to a solution of compound 5-6 (600.0mg) in tetrahydrofuran (15.0mL). The reaction solution was stirred at 25°C for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 0-50%) to obtain compound 5-7. LC-MS: m / z = 381.1 [M+H] + .

[0125] Step 5 At 20°C, under the protection of nitrogen gas, compound 5-7 (200.0 mg) was dissolved in methanol (1.0 mL) and concentrated H2SO4 (1.0 mL) was added. The reaction solution was heated to 80°C and stirred for 3 hours. The reaction solution was cooled to room temperature, diluted with water (50.0 mL), adjusted to pH 7-8 with saturated aqueous sodium bicarbonate solution, and extracted with ethyl acetate (50.0 mL x 2). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel thin layer chromatography. Compound 5-8 was obtained by separation and purification using a plate (dichloromethane:methanol, methanol ratio: 0-10%). LC-MS: m / z=310.2[M+H] + .

[0126] Step 6 Sodium borohydride acetate (137.0 mg) was added to 1,2-dichloroethane (1.0 mL) containing compound 5-8 (50.0 mg) and compound M-2 (51.4 mg) at 25°C. The mixture was stirred at 25°C for 16 hours. Water (200.0 mL) was added for dilution, and the mixture was extracted with ethyl acetate (50.0 mL x 2). The organic phases were combined, washed with water (100.0 mL x 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, ethyl acetate ratio: 0-15%) to obtain compound 5-9. LC-MS: m / z = 583.4 [M + H] + .

[0127] Step 7 At 25° C., compound 5-9 (30.0 mg) was dissolved in tetrahydrofuran (0.5 mL) and methanol (0.5 mL), lithium hydroxide monohydrate (13.0 mg) was added, and the reaction solution was stirred at 50° C. for 16 hours. The reaction solution was cooled to room temperature, quenched by adding acetic acid (0.1 mL), diluted with water (50.0 mL), extracted with ethyl acetate (25.0 mL×2), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure to obtain compound 4C. LC-MS: m / z=469.1 [M+H] + .

[0128] Example 6: Preparation of Compound 4D [ka]

[0129] Step 1 At 25°C, compound 5-1 (50.00g) and compound 6-2 (45.92g) were dissolved in 2-methyltetrahydrofuran (500.0mL), and tetraethoxytitanium (235.72g) was added in batches. The reaction solution was heated to 80°C under the protection of nitrogen gas and stirred for 16 hours. The reaction solution was poured into ice water (15.0L), filtered, the cake was washed with 2-methyltetrahydrofuran (3.0L), the filtrate was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove 2-methyltetrahydrofuran (400.0mL), and the remaining solution was poured into petroleum ether (200.0mL) at 0°C, stirred for 1 hour, filtered, the cake was collected, and dried under reduced pressure to obtain compound 6-3. 1 H NMR (400MHz, CDCl3 ) δ:7.95-7.97(d,J=8.0Hz,2H),7.72-7.74(d,J=8.0Hz,2H),2.79(s,3H),1.33(s,9H).

[0130] Step 2 At -20°C, under the protection of nitrogen gas, lithium diisopropylamide (2M tetrahydrofuran solution, 60.40mL) was added dropwise to a solution of compound 6-3 (25.00g) in 2-methyltetrahydrofuran (500.0mL), and after completion of the addition, the reaction solution was stirred at -20°C to -15°C for 1 hour, and a solution of compound 5-4 (35.35g) in 2-methyltetrahydrofuran (125.0mL) was added, the temperature was slowly raised, and the mixture was stirred at -20°C to -15°C for 4 hours. The reaction was quenched by adding a saturated aqueous ammonium chloride solution (60.00mL), diluted with water (300.0mL), and extracted with ethyl acetate (200.0mL x 2). The organic phases were combined, washed with water (200.0mL x 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, ethyl acetate ratio: 0 to 13%) to obtain compound 6-5.

[0131] Step 3 Compound 6-5 (5.20 g) was dissolved in tetrahydrofuran (52.0 mL) at 0°C, sodium borohydride (0.50 g) was added, and the reaction system was stirred at 0°C for 1 hour. Saturated aqueous ammonium chloride solution (50.0 mL) was added to quench the reaction, water (200.0 mL) was added to dilute the reaction, and the mixture was extracted with ethyl acetate (100.0 mL x 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, ethyl acetate ratio: 0-33%) to obtain compound 6-6. LC-MS: m / z = 441.1 [M + H] + .

[0132] Step 4 Compound 6-6 (3.30 g) was dissolved in tetrahydrofuran (15.0 mL) at -15°C, lithium triisobutylhydroborate (1 M tetrahydrofuran solution, 5.02 mL) was added, and the reaction system was stirred at -15°C for 4 hours. Saturated ammonium chloride solution (50.0 mL) was added to quench the reaction, water (200.0 mL) was added to dilute the reaction, and the mixture was extracted with ethyl acetate (50.0 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (dichloromethane:methanol, methanol ratio: 0-5%) to obtain compound 6-7. LC-MS: m / z = 399.2 [M + H] + .

[0133] Step 5 At 25°C, triphenylphosphine (5.30g) and diisopropyl azodicarboxylate (4.08g) were added to a solution of compound 6-7 (2.30g) in tetrahydrofuran (46.0mL). The reaction solution was stirred at 25°C under nitrogen gas protection for 16 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, ethyl acetate ratio: 0-40%) to obtain compound 6-8. LC-MS: m / z=381.1[M+H] + .

[0134] Step 6 At 20°C, under the protection of nitrogen gas, compound 6-8 (500.0 mg) was dissolved in methanol (5.0 mL) and concentrated H2SO4 (5.0 mL) was added. The reaction solution was heated to 80°C and stirred for 16 hours. The reaction solution was cooled to room temperature, the pH was adjusted to 7-8 with saturated aqueous sodium bicarbonate solution, and extracted with ethyl acetate (50.0 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was separated and purified on a silica gel thin layer chromatography plate (dichloromethane:methanol = 0-10%). Compound 6-9 was obtained. LC-MS: m / z=310.1 [M+H] + .

[0135] Step 7 Sodium borohydride acetate (356.3 mg) was added to 1,2-dichloroethane (3.0 mL) containing compound 6-9 (130.0 mg) and compound M-2 (133.7 mg) at 25°C. The mixture was stirred at 25°C for 16 hours. Water (200.0 mL) was added for dilution, and the mixture was extracted with ethyl acetate (50.0 mL x 2). The organic phases were combined, washed with water (100.0 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, ethyl acetate ratio: 0-15%) to obtain compound 6-10. LC-MS: m / z =583.4 [M+H] + .

[0136] Step 8 At 25°C, compound 6-10 (30.0 mg) was dissolved in tetrahydrofuran (0.5 mL) and methanol (0.5 mL), lithium hydroxide monohydrate (13.0 mg) was added, and the reaction solution was stirred at 50°C for 16 hours. The reaction solution was cooled to room temperature, quenched by adding acetic acid (0.1 mL), diluted with water (50.0 mL), extracted with ethyl acetate (25.0 mL x 2), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure to obtain compound 4D. LC-MS: m / z = 469.1 [M + H] + .

[0137] Biological Test Data: Experimental Example 1: Wieslab alternative complement pathway activation inhibition (enzyme activity test) The purpose of this experiment is to determine the inhibitory activity of the compounds of the present invention against the alternative complement pathway in human serum by using the Wieslab® alternative complement pathway kit.

[0138] Testing Method: Serum was diluted with diluent (1:23). Drug was added to the diluted serum and diluted 5-fold gradient with 8 concentration gradients up to 10mM or 50mM. Incubated at room temperature for 15 minutes. Compound and serum mixture was added to 96-well plate (100μL / well) provided with the kit and activated at 37℃ for 1 hour. Washed 3 times with wash buffer. Detection antibody provided with the kit (100μL) was added and incubated at room temperature for 30 minutes. Washed 3 times with wash buffer. Substrate (100μL) was added and incubated at room temperature for 30 minutes. Absorbance at 405nM was detected with a microplate reader.

[0139] Experimental results: The inhibitory activity of the test compounds against the alternative complement pathway is shown in Table 1. [Table 1] Conclusion: The compounds of the present invention have significant inhibitory activity against the activation of the bypass pathway in human serum. has.

[0140] Experimental Example 2: Pharmacokinetic study in mice Study objective: To detect the in vivo plasma pharmacokinetics of the compounds of the present invention after a single intravenous injection and intragastric administration in male C57BL / 6J mice.

[0141] Experimental animals: male C57BL / 6J mice, 7-9 weeks old, weight: 17-23g; Supplier: Shanghai Sippe-Bk Lab Animal Co.,Ltd.

[0142] Experimental process: Injection (IV): Dose: 1 mg / kg (Vehicle: 10% PG / 5% Solutol / 85% PBS (1x pH 7.4)); Oral (PO): Dose: 10 mg / kg (Vehicle: 30% PEG300 / 10% Cremphor EL / 60% PBS (1x pH 7.4)).

[0143] Sample collection: 0.03 mL blood samples were collected by saphenous vein puncture of experimental animals at each time point, and the actual blood collection time was recorded. All blood samples were placed in 1.5 mL commercial EDTA-K2 anticoagulant tubes (supplier: Jiangsu Kangjian Medical Apparatus Co., Ltd.). Within 30 min after blood samples were collected, they were centrifuged at 3000 g for 10 min at 4°C to aspirate the supernatant plasma, which was then immediately placed on dry ice and stored in a -80°C refrigerator for use in LC-MS / MS analysis.

[0144] Data analysis: WinNonlin TM Plasma concentrations were processed using a noncompartmental model in Version 6.3 (Pharsight, Mountain View, CA) pharmacokinetic software, and the pharmacokinetic parameters Cl, T were calculated by the linear-logarithmic ladder method. 1 / 2 , C max , AUC 0-last was calculated, and the results are shown in Table 2. [Table 2]

[0145] Experimental Conclusion: The results of the pharmacokinetic study of the compound of the present invention in mice show that it has a longer half-life than the reference compound and a significantly higher oral plasma exposure than the reference compound LNP023, indicating that the compound of the present invention has good pharmacokinetic properties.

[0146] Experimental Example 3: Pharmacokinetic study in rats Test objective: To detect the in vivo plasma pharmacokinetics of the compounds of the present invention after a single intravenous injection and intragastric administration in male Wistar Han mice.

[0147] Experimental animals: male Wistar Han rats, 7-9 weeks old, weight: 231.03-243.2g; Supplier: Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.

[0148] Experimental process: Injection (IV): Dose: 1mpk (Vehicle: 30% PEG300 / 10% Cremphor EL / 60% PBS (1x pH 7.4)); Oral (PO): Dose: 5mpk or 30mpk (Vehicle: 30% PEG300 / 10% Cremphor EL / 60% PBS (1x pH 7.4)).

[0149] Sample collection: Approximately 0.3 mL of blood samples were collected by jugular vein puncture of the experimental animals at each time point, and the actual blood collection time was recorded. All blood samples were placed in 1.5 mL commercial EDTA-K2 anticoagulant tubes (supplier: Jiangsu Kangjian Medical Apparatus Co., Ltd.). After blood samples were collected, they were centrifuged at 3200 g at 4°C for 10 min to aspirate the supernatant plasma, which was then immediately placed on dry ice and stored in a -80°C refrigerator for use in LC-MS / MS analysis.

[0150] Data analysis: Plasma concentrations were processed using a noncompartmental model in Phoenix WinNonlin 6.3 pharmacokinetic software, and the pharmacokinetic parameters Cl, C were calculated by the linear-logarithmic ladder method. max , T 1 / 2 , AUC 0-last was calculated, and the results are shown in Table 3. [Table 3]

[0151] Experimental Conclusion: The results of the pharmacokinetic study of the compound of the present invention in rats show that it has a longer half-life than the reference compound and a significantly higher oral plasma exposure than the reference compound LNP023, indicating that the compound of the present invention has good pharmacokinetic properties.

[0152] Experimental example 4: In vivo PD model of LPS-induced complement activation in mice Experimental objective: to detect the inhibitory effect of the compound of the present invention on the activation of complement in mice induced by LPS stimulation.

[0153] Experimental animals: female C57BL / 6J mice, 7-9 weeks old, weight: 17-23g; Supplier: Shanghai Sippe-Bk Lab Animal Co.,Ltd.

[0154] Experimental process: Complement activation was induced in mice by intraperitoneal injection of 100 μg lipopolysaccharide (LPS) in Salmonella typhimurium (Sigma) in 100 μL sterile PBS. Negative control animals were injected intraperitoneally with 100 μL sterile PBS and administered alone by gavage (PO). Positive control animals were injected intraperitoneally with LPS and PO drug vehicle (0.5% (w / v) methylcellulose and 0.5% (v / v) Tween 80). The administration and plasma collection times were as shown below: TIFF0007641388000055.tif25146

[0155] Sample collection: 0.3 mL of blood was collected from the orbital venous plexus. All blood samples were placed in 1.5 mL commercial EDTA-K2 anticoagulant tubes (supplier: Jiangsu Kangjian Medical Apparatus Co., Ltd.). Within 30 minutes of collection, blood samples were centrifuged at 3000 g for 10 minutes at 4°C to aspirate the supernatant plasma, which was then immediately placed on dry ice and stored in a -80°C refrigerator for use in the analysis of downstream C3d protein levels after complement activation by Westernblot.

[0156] Sample analysis: Mouse plasma (5 μL) + lysis buffer (27.5 μL) + loading buffer (12.5 μL) + reducing buffer (5 μL) were mixed uniformly and incubated at 100°C for 15 minutes. The loading amount was 5 μL / well, i.e., the loading amount of plasma in each well was 0.5 μL.

[0157] Experimental results: In this experiment, Novartis Factor B inhibitor LNP023 was used as the reference drug, and the test compounds were reference compound LNP023, compound 3A and compound 3C. All three drugs could significantly inhibit the level of C3d, that is, inhibit complement activation induced by LPS, and the inhibitory effect of compound 3A and compound 3C was better than that of LNP023. The specific experimental results are shown in Figure 1, and the data are expressed as follows: Mean ± SEM, n = 5; ###, p < 0.001, normal group vs. model group, t-test; ****, p < 0.0001, treatment group vs. model group, one-way ANOVA.

[0158] Experimental conclusion: The compounds of the present invention can significantly inhibit LPS-stimulated complement activation, and the inhibitory effect is better than that of LNP023.

[0159] Experimental example 5: In vivo pharmacodynamic model of passive Heymann nephritis in rats Experimental Objective: To investigate the ability of the compounds of the present invention to improve renal function in rats with Heymann nephritis induced by Sheep Anti-Rat Fx1A Serum (Goat Anti-Mouse Fx1A Serum), including evaluating the reduction of proteinuria levels and the improvement of renal tissue damage.

[0160] Experimental animals: male SD rats, 7-10 weeks old, weight: 200-300g; Supplier: Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.

[0161] Experimental process: 1. Modeling: On day D-2, before administration, rat urine was collected. On day D1, the first day of the experiment, animals in the control group (group 1) were injected once with 5mL / kg of sheep non-immune serum via the tail vein, and animals in the model group and administration groups (groups 2-6) were injected once with 5mL / kg of sheep anti-rat Fx1A serum via the tail vein.

[0162] 2. Dosage: The administration method was intragastric administration twice a day with an 8-hour administration interval, and the administration volume was 10mL / kg. On day D1, 1 hour before modeling, the animals in groups 1 and 2 were administered blank solvent 20% PEG400 / 10% Solutol / 70% water, the animals in group 3 were administered LNP023 (60mpk), and the animals in groups 4 to 6 were administered different concentrations of compound 4B (5mpk, 20mpk and 60mpk), and 8 hours after the first administration interval, each group was administered again with the same dose and administration volume as the first. On days D2 to D14, the animals in each group were administered different compounds or solvents continuously for 14 days (including day 1) according to the dose, volume, administration method and frequency on day D1.

[0163] 3. Sample collection: Urine collection: On D~2 days before administration, and on D4, D6, D8, D11, and D14 after administration, rat urine was collected 2~4 hours and 4~6 hours after the first administration, transferred to EP tubes, and stored in a refrigerator at -80℃~-60℃ for use in the detection of rat urinary protein and creatinine.

[0164] Kidney sample collection: All animals were euthanized by CO2 inhalation anesthesia on day 15, and bilateral kidneys were harvested. The left kidney was cut transversely and the right kidney was cut longitudinally, and the transverse half (left side) and longitudinal half (right side) were fixed in formalin (in the same EP tube).

[0165] 4. Sample analysis: (1) Rat urinary creatinine uCRE was usually diluted 10 times (Decrease mode), 10 μL sample + 90 μL saline (0.9% saline), if it exceeded the upper limit of detection, it was diluted 20 times (Increase mode), 5 μL sample + 95 μL saline (0.9% saline); rat urinary total protein uTP was detected at the original multiple, if it exceeded the upper limit of detection, it was first diluted 10 times (Decrease mode), 10 μL sample + 90 μL saline (0.9% saline), if it still exceeded the upper limit of detection after 10 times dilution, it was diluted 100 times (Increase mode), 2 μL sample + 198 μL saline (0.9% saline); data was read by the analytical instrument HITACHI LST008 AS(P).

[0166] (2) Renal pathology score: HE staining was used to analyze the degree of drug damage to rat kidneys (paraffin sections). Score criteria: 0 means normal, 1 means slight cell infiltration in the mesangium, 2 means more cell infiltration in the mesangium, 3 means some glomerular mesangial cells have proliferated and some mesangial cells have infiltrated, and 4 means tubular atrophy, formation and sclerosis of glomerular crescents.

[0167] Experimental results: Novartis Factor B inhibitor LNP023 was used as the reference drug in the detection of urinary protein and creatinine in rats. Both LNP023 and compound 4B could significantly inhibit the urinary protein level of rats and improve the renal function of rats, and compound 4B was more effective than LNP023 at a dose of 60mpk. The specific experimental results are shown in Figure 2, and the data are expressed as follows: . Mean±SEM, n=5;###, p<0.001, normal group vs. model group, t-test; ****, p<0.0001, treated group vs. model group, one-way ANOVA.

[0168] The pathological score results are shown in Table 4. [Table 4]

[0169] Experimental conclusion: According to the pathological score results, compared with the model group, the compounds of the present invention in different dosage groups can significantly improve the degree of renal lesions in model animals. The compounds of the present invention can reduce the urinary protein level of rats and improve renal function, and are more effective than LNP023.

Claims

1. A compound represented by formula (I) or a pharma- ceutically acceptable salt thereof. 【Chemistry 1】 (however, L is a single bond or NR ４ is selected from R １ is C ３－６ Cycloalkyl, 3- to 6-membered heterocycloalkyl, -C １－３ Alkyl-C １－３ Alkoxy, -C １－３ Alkyl-C ３－６ Cycloalkyl and -C １－３ alkyl-3 to 6 membered heterocycloalkyl; ３－６ Cycloalkyl, 3- to 6-membered heterocycloalkyl, -C １－３ Alkyl-C １－３ Alkoxy, -C １－３ Alkyl-C ３－６ Cycloalkyl and -C １－３ Alkyl-3 to 6-membered heterocycloalkyl is independently selected from the group consisting of 1, 2 or 3 R ａ is replaced by R ２ , R ３ and R ４ are each independently H and C １－５ alkyl, １－５ The alkyl group may optionally have one, two or three R ｂ is replaced by Each R ａ and R ｂ are each independently selected from H, F, Cl, Br and I.

2. The compound according to claim 1, which is a compound represented by the following formula (I-1) or a pharma- ceutically acceptable salt thereof: 【Chemistry 2】 (however, L, R １ , R ２ and R ３ is as defined in claim 1, Carbon atoms marked with an "*" are chiral carbon atoms and exist in either (R) or (S) single enantiomer form or in (R) or (S) enantiomerically enriched form.

3. The compound according to claim 2, which is a compound selected from the group consisting of the following formulas (I-1-1), (I-1-2), (I-1-3) and (I-1-4), or a pharma- ceutically acceptable salt thereof. 【Chemistry 3】 (However, L, R １ , R ２ and R ３ is as defined in claim 2.

4. R of formula (I), formula (I-1), formula (I-1-1), formula (I-1-2), formula (I-1-3) or formula (I-1-4) １ is based on the following formula: 【Chemistry 4】 and the above group is selected from 【Chemistry 5】 are each independently selected from optionally one, two or three R ａ The compound according to any one of claims 1 to 3, or a pharma- ceutically acceptable salt thereof, which is substituted by:

5. R of formula (I), formula (I-1), formula (I-1-1), formula (I-1-2), formula (I-1-3) or formula (I-1-4) １ is based on the following formula: 【Chemistry 6】 5. The compound of claim 4, selected from:

6. R of formula (I), formula (I-1), formula (I-1-1), formula (I-1-2), formula (I-1-3) or formula (I-1-4) ２ H and CH ３ wherein said CH ３ optionally 1, 2 or 3 R ｂ The compound according to any one of claims 1 to 3, or a pharma- ceutically acceptable salt thereof, which is substituted by:

7. R of formula (I), formula (I-1), formula (I-1-1), formula (I-1-2), formula (I-1-3) or formula (I-1-4) ２ But, H, CH ３ , C.F. ３ and CHF ２ 7. The compound of claim 6, selected from:

8. R of formula (I), formula (I-1), formula (I-1-1), formula (I-1-2), formula (I-1-3) or formula (I-1-4) ３ H and CH ３ The compound according to any one of claims 1 to 3, or a pharma- ceutically acceptable salt thereof, selected from:

9. R of the formula (I), formula (I-1), formula (I-1-1), formula (I-1-2), formula (I-1-3) or formula (I-1-4) ４ H and CH ３ The compound according to any one of claims 1 to 3, or a pharma- ceutically acceptable salt thereof, selected from:

10. The compound or a pharma- ceutically acceptable salt thereof according to any one of claims 1 to 3, wherein L in the formula (I), (I-1), (I-1-1), (I-1-2), (I-1-3) or (I-1-4) is a single bond.

11. A partial structure in the formula (I). 【Chemistry 7】 2. The compound according to claim 1, wherein: 【Chemistry 8】

12. A partial structure in the formula (I). 【Chemistry 9】 12. The compound according to claim 11, wherein: 【Chemistry 10】

13. A compound represented by the following formula or a pharma- ceutically acceptable salt thereof: 【Chemistry 11】

14. 14. The compound of claim 13 selected from the following formulas: or a pharma- ceutically acceptable salt thereof. 【Chemistry 12】 15. Use of a compound according to any one of claims 1 to 14 or a pharma- ceutically acceptable salt thereof in the manufacture of a medicament for treating an immune disease.

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