Series of piperidine substituted benzoic acid compounds and application thereof

Abstract

The invention discloses a series of piperidine substituted benzoic acid compounds and application thereof, and particularly discloses a compound as shown in a formula (I) and pharmaceutically acceptable salts thereof.

Description

[0001] This patent application is a divisional application of patent application No. 2021800844732, the filing date of which is December 30, 2021, and the title of which is "A series of piperidine-substituted benzoic acid compounds and applications thereof". TECHNICAL FIELD

[0002] The present application relates to a series of piperidine-substituted benzoic acid compounds, in particular to a compound represented by formula (I) and a pharmaceutically acceptable salt thereof. BACKGROUND

[0003] Immune diseases are diseases caused by the imbalance of immune regulation affecting the immune response of the body. The complement system is an important component of the immune system, which includes a group of proteins that usually exist in an inactive state, i.e., complement. Complement can be activated in the alternative activation pathway by substances such as lipopolysaccharide, polysaccharide, peptidoglycan, teichoic acid, and agglutinated IgA and IgG4, mediating immune response and inflammatory response. Among them, the activator directly activates C3 and then completes the cascade reaction of each component from C5 to C9. Complement factor B (complement factor B), also known as C3 activator precursor, can be cleaved into Ba and Bb fragments by complement factor D, and Bb combines with C3b to form C3 convertase to play a role in the alternative pathway. Complement Factor B acts on the AP pathway, and inhibition of Factor B activity can prevent API pathway activation without interfering with CP and LP pathways, which can avoid the increased risk of infection due to complement system inhibition.

[0004] Currently, there is no small molecule Factor B inhibitor on the market. Novartis' factor B inhibitor LNP023 is in clinical phase III research for the treatment of PNH, IgAN, C3G and other diseases. Therefore, there is an urgent need in the art to develop new complement system Factor B small molecule inhibitors to increase clinical research and verification and for the treatment of various diseases caused by complement abnormalities. SUMMARY

[0005] The present application provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

[0006]

[0007] wherein,

[0008] L is selected from a single bond, NR4, and O;

[0009] R1is selected from fluorine, chlorine, C 1-5 alkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, -C 1-3 alkyl-C1-3 alkyl-C 1-3 alkyl-C 3-6 cycloalkyl and -C 1-3 alkyl-3-6 membered heterocycloalkyl, said 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-3-6 membered heterocycloalkyl, each independently optionally substituted with 1, 2, or 3 R a substituents;

[0010] R2, R3and R4are each independently selected from H and C 1-5 alkyl, said C 1-5 alkyl is optionally substituted with 1, 2, or 3 R b substituents;

[0011] each R a and R b is each independently selected from H, F, Cl, Br, and I;

[0012] provided that when R1is selected from unsubstituted C 1-5 alkyl, R2and R3are not simultaneously selected from H.

[0013] In some embodiments of the present application, the compound or a pharmaceutically acceptable salt thereof is selected from,

[0014]

[0015] wherein,

[0016] L, R1, R2and R3are as defined in the present application.

[0017] The carbon atom with “*” is a chiral carbon atom, which exists in the form of (R) or (S) single enantiomer or enriched in one enantiomeric form.

[0018] In some embodiments of the present application, the compound or a pharmaceutically acceptable salt thereof is selected from,

[0019] and ,

[0020] wherein, L, R1, R2and R3are as defined in the present application.

[0021] In some embodiments of the present application, the R1is selected from C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, -C1-3 Alkyl-C 1-3 Alkoxy, -C 1-3 Alkyl-C 3-6 cycloalkyl and -C 1-3 Alkyl-3-6-membered heterocyclic alkyl, wherein the 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-3-6-membered heterocyclic alkyl groups are each independently and optionally surrounded by 1, 2, or 3 R groups. a Replacement, other variables as defined in this invention.

[0022] In some embodiments of the present invention, R1 is selected from fluorine, chlorine, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, , , , , , and The CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, , , , , , and Each can be independently selected by 1, 2 or 3 Rs. a Replacement, other variables as defined in this invention.

[0023] In some embodiments of the present invention, R1 is selected from CH3, CF3, CH2CH3, CH2CHF2, CH2CF3, , , , , , and Other variables are as defined in this invention.

[0024] In some embodiments of the present invention, R1 is selected from... , , , , , and Other variables are as defined in this invention.

[0025] In some embodiments of the application, said R1is selected from C 1-5 alkyl, R2is selected from H and C 1-5 alkyl, R3is selected from C 1-5 alkyl, other variables are as defined in the application.

[0026] In some embodiments of the application, said R1is selected from CH2CH3, R2is selected from H and CH3, R3is selected from CHF2and CH3, other variables are as defined in the application.

[0027] In some embodiments of the application, said R2is selected from H and CH3, said CH3is optionally substituted with 1, 2 or 3 R b , other variables are as defined in the application.

[0028] In some embodiments of the application, said R2is selected from H, CH3and CHF2, other variables are as defined in the application.

[0029] In some embodiments of the application, said R3is selected from H and CH3, other variables are as defined in the application.

[0030] In some embodiments of the application, said R4is selected from H and CH3, other variables are as defined in the application.

[0031] In some embodiments of the application, said L is selected from a single bond and O, other variables are as defined in the application.

[0032] In some embodiments of the application, said structural unit is selected from , , , , , , , , , , , and , other variables are as defined in the application.

[0033] In some embodiments of the application, said structural unit is selected from , , , , , , , , , , , , and , with the other variables being as defined herein.

[0034] In some embodiments of the application, the structural unit is selected from , , , , , , , , , , , , , , , , , , , , , , , , , , , , and , with the other variables being as defined herein.

[0035] The present application provides a compound of Formula (I), or a pharmaceutically acceptable salt thereof,

[0036]

[0037] wherein,

[0038] L is selected from a single bond, NR4, and O;

[0039] R1is selected from fluoro, chloro, 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-3-6 membered heterocycloalkyl, said 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 1-3alkyl-3-6 membered heterocycloalkyl is independently optionally substituted by 1, 2 or 3 R a substituted;

[0040] R2, R3and R4are each independently selected from H and C 1-5 alkyl, said C 1-5 alkyl is optionally substituted by 1, 2 or 3 R b substituted;

[0041] each R a and R b are each independently selected from H, F, Cl, Br and I;

[0042] with the proviso that when R1is selected from C 1-5 alkyl, R2and R3are not simultaneously selected from H.

[0043] The present application also provides for certain embodiments of the application wherein the compound is selected from the group consisting of:

[0044] The present application also provides for certain embodiments of the application wherein the compound is selected from the group consisting of:

[0045] , , , , , , , , , , , , and .

[0046] The present application also provides for certain embodiments of the application wherein the compound is selected from the group consisting of:

[0047]

[0048]

[0049]

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056]

[0057]

[0058]

[0059]

[0060]

[0061] and .

[0062] The application also provides application of the compound or the pharmaceutically acceptable salt thereof in preparation of a drug related to complement factor B.

[0063] The application also provides the following test methods:

[0064] Method 1. PHN Heymann nephritis efficacy test

[0065] 1.1 Purpose of the experiment: to investigate the renal function improvement ability of the compound of the application on sheep anti-rat Fx1A serum induced rat Heymann nephritis, including the evaluation of reducing the proteinuria level and improving the kidney tissue damage.

[0066] 1.2 Experimental animals: male SD rats, 7-10 weeks old, weighing 200-300 grams

[0067] 1.3 Experimental process:

[0068] 1.3.1 Modeling: on D-2 days before administration, collect the urine of rats. D1 day is the first day of the experiment, the control group (group 1) animals are given a single injection of 5 mL / kg of Sheep Non-Immune serum through the tail vein; the modeling group and the administration group (groups 2-6) animals are given a single injection of 5 mL / kg of Sheep Anti-Rat Fx1A Serum through the tail vein.

[0069] 1.3.2 Drug administration:

[0070] The administration mode is intragastric administration, twice a day, with an administration interval of 8 hours, and the administration volume is 10 mL / kg. On D1, 1 hour before modeling, the animals in groups 1 and 2 are given blank solvent 20% PEG 400 / 10% Solutol / 70% water; the animals in group 3 are given LNP023 (60 mpk); the animals in groups 4-6 are given compound 4B at different concentrations (5 mpk, 20 mpk and 60 mpk); 8 hours after the first administration, each group is administered again, with the same dose and volume as the first administration. On D2-D14, the animals in each group are continuously administered (including the first day) with different compounds or solvents according to the dose, volume, administration mode and frequency of D1.

[0071] 1.3.3 Sample collection:

[0072] (1) Urine collection: On D-2 days before administration, and on D4, D6, D8, D11, D14 days, 2-4 h and 4-6 h after the first administration, urine samples of rats are collected into EP tubes and stored in a refrigerator at -80℃ to -60℃ for detection of rat urine protein and urine creatinine.

[0073] (2) Kidney tissue collection: All animals are euthanized by CO2 inhalation anesthesia on D15 days after administration, and bilateral kidneys are collected. The left kidney is cut transversely, and the right kidney is cut longitudinally. The transversely cut half (left) and the longitudinally cut half (right) are placed in formalin fixation (in the same EP tube), and the remaining transversely cut half (left) and the longitudinally cut half (right) are OCT embedded, with the cut surface facing down (in the same embedding box). After embedding, they are stored as soon as possible in a refrigerator at -80℃ to -60℃ for histopathological scoring.

[0074] 1.3.4 Sample analysis: Urine protein and urine creatinine data are read by the analyzer HITACHI LST008 AS(P). The kidney glomerular disease (characterized by glomerular enlargement, basement membrane and Bowman's capsule thickening, and podocyte enlargement / rounding) is scored by histopathological analysis, the tubular degeneration score is determined by histopathological analysis, and the glomerular C3 deposition score is analyzed by IHC or IF staining.

[0075] Technical effects

[0076] The compound of the present application has obvious inhibition activity on human serum bypass pathway activation, can significantly inhibit LPS-stimulated complement activation, reduce urine protein level, and improve kidney function; the PK results show that the compound of the present application has a long half-life and high drug exposure, excellent in vivo pharmacokinetic properties, and has excellent drug properties.

[0077] Definitions and explanations

[0078] The following terms and phrases, as used herein, are intended to have the following meanings unless otherwise indicated. A particular term or phrase should not be construed as undefined or unclear without a specific definition, but should be interpreted according to the ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commercial product or its active ingredient.

[0079] The term "pharmaceutically acceptable," as used herein, pertains to compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0080] The term "pharmaceutically acceptable salt" means a salt of a compound of the present application that is within the scope of sound medical judgment, of a compound of the present application having particular substituents discovered in the present application, with a relatively nontoxic acid or base. When a compound of the present application contains a relatively acidic moiety, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base to produce the salt. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or like salts. When a compound of the present application contains a relatively basic moiety, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid to produce the salt. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids, such as hydrochloric, hydrobromic, nitric, carbonic, bicarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, hydro sulfuric, hydriodic, phosphorous, and the like; and organic acids, such as acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, ethanesulfonic, and the like; also salts of amino acids such as arginate, gluconate, and the like; and salts of organic acids like glucuronic, and the like. Certain specific compounds of the present application contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0081] The pharmaceutically acceptable salts of the present application can be synthesized from the parent compound that contains an acidic or basic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of both. Generally, non-aqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

[0082] The compounds of the present application can exist in particular geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis- and trans-forms, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)-isomers, as well as racemic mixtures and other mixtures thereof, e.g., mixtures of enantiomers or diastereomers, all of which are intended to be within the scope of the present application. Additional asymmetric carbon atoms can be present in a substituent group such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be within the scope of the present application.

[0083] The term "enantiomeric" or "optical isomer" means a stereoisomer which is a mirror image of the other and which is not superimposable.

[0084] The term "cis" or "geometric isomer" means a stereoisomer which results from a double bond or a ring-forming carbon atom single bond which cannot rotate freely.

[0085] The term "diastereomeric" means a stereoisomer which has two or more chiral centers and which is not a mirror image of the other.

[0086] "(+)" means dextrorotary, "(-)" means levorotary, and "(±)" means racemic, unless otherwise indicated.

[0087] Unless otherwise indicated, a wedged solid line bond ( ) and a wedged dashed line bond ( ) represent the absolute configuration of a stereocenter, a straight solid line bond ( ) and a straight dashed line bond ( ) represent the relative configuration of a stereocenter, a wavy line ( ) represents either a wedged solid line bond ( ) or a wedged dashed line bond ( ), or a wavy line ( ) represents either a straight solid line bond ( ) or a straight dashed line bond ( ).

[0088] The term "tautomer" or "tautomer form" means, unless otherwise indicated, that different functional group isomers are in dynamic equilibrium at room temperature and can rapidly interconvert. If tautomers are possible (e.g., in solution), a chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also known as prototropic tautomers) include interconversions by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions by reorganization of some of the bonding electrons. A specific example of keto-enol tautomerization is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0089] The term "enantiomerically enriched," "enantiomeric excess," "enantiomeric excess," or "enantiomeric excess" means, unless otherwise indicated, that one enantiomer is present in an amount less than 100% and greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0090] The term "enantiomeric excess" or "enantiomeric excess" means, unless otherwise indicated, the difference between the relative percentages of the two enantiomers. For example, where one enantiomer is present in an amount of 90% and the other enantiomer is present in an amount of 10%, the enantiomeric excess (ee value) is 80%.

[0091] The compounds of the present application can contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds can be radiolabeled with radioactive isotopes, such as tritium ( 3 H), iodine-125 ( 125 I), or carbon-14 ( 14 C). For example, deuterium can be substituted for hydrogen to form deuterated drugs, which have advantages over non-deuterated drugs, such as reduced toxicity, increased stability, increased efficacy, increased biological half-life, etc. All isotopic variations of the compounds of the present application, whether radioactive or not, are encompassed within the scope of the present application.

[0092] The term "optionally" or "optional" means that the subsequently described event or circumstance may or can not occur, and this description includes instances where the event or circumstance occurs and instances where it does not.

[0093] The term "substituted" means that any one or more hydrogen atoms on a particular atom is / are replaced with a substituent group, which can include isotopes of hydrogen and variations of hydrogen, as long as the valency of the particular atom is normal and the resulting compound is stable. When the substituent group is oxygen (i.e., =0), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that the group can or can not be substituted, and unless otherwise specified, the kinds and number of substituents are any that are chemically possible.

[0094] When any variable (e.g., R) occurs more than one time in a compound or substituent, its definition in each instance is independent of the definition of the other. Thus, for example, if a group is substituted with 0-2 R groups, then the group is optionally substituted with up to two R groups, and the R group is independently selected at each occurrence. Also, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0095] When the number of occurrences of a linking group is zero, such as -(CRR)0-, it means that the linking group is a single bond.

[0096] When one of the variables is selected from a single bond, it means that the two groups to which it is attached are directly connected, such as when L represents a single bond in A-L-Z, the structure is actually A-Z.

[0097] When the listed linking groups do not indicate their direction of attachment, their direction of attachment is arbitrary, such as where the linking group L is -M-W-, then -M-W- can either connect ring A and ring B to form or ring A and ring B to form The combinations of the linking groups, substituents and / or variables are permissible only if such combinations result in stable compounds.

[0098] Unless otherwise specified, when a group has one or more attachable sites, any one or more of the sites of the group can be attached to other groups by a chemical bond. When the chemical bond is not directional and there is an H atom at the attachable site, the number of H atoms at the site is reduced by the number of chemical bonds attached to it to become a group of the corresponding valence. The chemical bond that attaches the site to other groups can be represented by a straight, solid line bond ), straight dashed key ( ), or wavy lines ( () indicates that the oxygen atom in the group is bonded to another group. For example, a straight solid line bond in -OCH3 indicates that the oxygen atom in the group is bonded to another group. The straight dashed bond in the diagram indicates that the group is connected to other groups through both ends of the nitrogen atom in the group; The wavy lines in the text indicate that the phenyl group is connected to other groups through the carbon atoms at positions 1 and 2 of the phenyl group. This indicates that any connectable site on the piperidinyl group can be linked to other groups via a single chemical bond, including at least... , , , Even if H atoms are drawn on -N- in these four connection methods, Still includes In this type of linkage, when a chemical bond is attached, the number of hydrogen atoms at that site is reduced by one, resulting in a monovalent piperidinyl group.

[0099] Unless otherwise specified, the term "C" 1-3 "alkyl" on its own or in combination with other terms refers to a straight-chain or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). C 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), etc.

[0100] Unless otherwise specified, the term "C" 1-5 "alkyl" on its own or in combination with other terms refers to a straight-chain or branched saturated hydrocarbon group consisting of 1 to 5 carbon atoms. The C 1-5 Alkyl groups include C 1-4 C 1-3 C 1-2 C 2-5 C 2-4 And C5 alkyl groups, etc.; they can be monovalent (such as methyl), divalent (such as methylene), or polyvalent (such as methine). C 1-5 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), etc.

[0101] Unless otherwise specified, the term "C" 1-3"Alkoxy" on its own, or in combination with other terms, refers to alkyl groups comprising 1 to 3 carbon atoms that are attached to the remainder of the molecule by an oxygen atom. 1-3 Alkoxy groups include C 1-2 C 2-3 C3 and C2 alkoxy groups, etc. 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), etc.

[0102] Unless otherwise specified, "C 3-6 "Cycloalkyl" on its own or in combination with other terms respectively represents a saturated monocyclic cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, wherein the C 3-6 Cycloalkyl groups include C 3-5 C 4-5 and C 5-6 Cycloalkyl groups, etc.; they can be monovalent, divalent, or polyvalent. C 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0103] Unless otherwise specified, the term "3-6 membered heterocyclic alkyl" on its own or in combination with other terms respectively refers to a saturated monocyclic cyclic group consisting of 3 to 6 ring atoms, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms independently selected from O, S and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e. NO and S(O)). p (where p is 1 or 2). Furthermore, regarding the "3-6 membered heterocyclic alkyl", the heteroatom can occupy the connection position between the heterocyclic alkyl and the rest of the molecule. The 3-6 membered heterocyclic alkyl includes 4-6, 5-6, 4, 5, and 6 membered heterocyclic alkyls, etc. Examples of 3-6 membered heterocyclic alkyl groups include, but are not limited to, azirrobutyl, oxacyclobutyl, thiocyclobutyl, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiopheneyl (including tetrahydrothiophene-2-yl and tetrahydrothiophene-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxyl, dithiaalkyl, isoxazolyl, isothiazolyl, 1,2-oxazinyl, 1,2-thiaazinyl or hexahydropyridazinyl, etc.

[0104] Unless otherwise specified, C n-n+m Or C n -C n+m This includes any specific case with n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C10 , C 11 , and C 12 , also includes any range of n to n+m, for example C 1-12 includes C 1-3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12 , etc.; similarly, n-membered to n+m-membered means the number of atoms in the ring is n to n+m, for example 3-12 membered ring includes 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, 7-membered ring, 8-membered ring, 9-membered ring, 10-membered ring, 11-membered ring, and 12-membered ring, also includes any range of n to n+m, for example 3-12 membered ring includes 3-6 membered ring, 3-9 membered ring, 5-6 membered ring, 5-7 membered ring, 6-7 membered ring, 6-8 membered ring, and 6-10 membered ring, etc.

[0105] The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, embodiments formed using combinations of the other chemical synthetic methods well known to those skilled in the art, and equivalents thereof as appreciated by those skilled in the art, preferred embodiments including, but not limited to, the examples of the present application.

[0106] The solvents used in the present application are commercially available.

[0107] The following abbreviations are used in the present application: aq stands for water; eq stands for equivalent, equivalents; DCM stands for dichloromethane; PE stands for petroleum ether; DMSO stands for dimethyl sulfoxide; 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-butyloxycarbonyl, an amine protecting group; r.t. 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 palladium tetrakis(triphenylphosphine); Pd(dppf)Cl2 .CH2Cl2represents [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex; Pd(dppf)Cl2represents [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium; PPA represents polyphosphoric acid; NMP represents N-methylpyrrolidine; TBSCl represents tert-butyldimethylsilyl chloride; n-BuLi represents n-butyllithium; TBAF represents tetra-n-butylammonium fluoride; psi represents pounds per square inch; CO2represents carbon dioxide; DEA represents diethylamine; PEG300 represents polyethylene glycol 300; Cremphor EL represents polyoxyl castor oil; PBS represents phosphate buffered saline.

[0108] The compounds of the present application can be confirmed by conventional methods well known to those skilled in the art. If the present application relates to the absolute configuration of a compound, the absolute configuration can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction method (SXRD), the grown single crystal is collected by Bruker D8 venture diffractometer to collect diffraction intensity data, light source is Cu Kα radiation, scanning mode: φ / ω scanning, after collecting relevant data, further using direct method (Shelxs97) to analyze crystal structure, the absolute configuration can be confirmed. BRIEF DESCRIPTION OF DRAWINGS

[0109] Figure 1 : Administration and plasma collection time points in Example 4;

[0110] Figure 2 : Experimental results of PD model in vivo of LPS-induced complement activation mice;

[0111] Figure 3 : Experimental results of passive Heymann nephritis rats in vivo. DETAILED DESCRIPTION

[0112] The present application is described in detail below by way of Examples, but it is not meant to be limited by any of the Examples. The present application has been described in detail and specific embodiments thereof have been disclosed herein, but it is to be understood that the detailed description and specific embodiments thereof which have been disclosed are illustrative only and that various changes and modifications can be suggested to one skilled in the art, without departing from the spirit and scope of the present application.

[0113] Preparation of Reference Example 1 intermediates M-2 and M-1

[0114]

[0115] First step

[0116] To a solution of compound M-A (5.00 g) in acetonitrile (30.0 mL) was added 4-dimethylaminopyridine (3.79 g) and di-tert-butyl dicarbonate (8.12 g) at 20 °C. The reaction solution was stirred at 20 °C for 1 h, diluted with ethyl acetate (100.0 mL), washed with 1 N hydrochloric acid (30.0 mL), saturated brine (50.0 mL*2) successively, dried over anhydrous sodium sulfate, filtered, and concentrated to give compound M-B. 1 H NMR (400 MHz, CDCl3) δ 7.52 (d, J =4.0 Hz, 1H), 6.87 (d,J =2.4 Hz, 1H), 6.75 (d, J =2.4 Hz, 1H), 6.48 (d, J =4.0 Hz, 1H), 3.85 (s,3H), 2.64 (s, 3H), 1.65 (s, 9H)。

[0117] Second step

[0118] To a solution of N-methylformanilide (5.59 g) in dichloromethane (50.0 mL) was added oxalyl chloride (5.25 g) at 20 °C. The reaction solution was stirred at 20 °C for 14 h under nitrogen atmosphere. The reaction solution was transferred into a constant pressure dropping funnel, and was slowly added to a solution of compound M-B (9.00 g) in dichloromethane (50.0 mL) at -14 °C. The reaction solution was stirred at -14 °C for 3 h under nitrogen atmosphere. The reaction was quenched by adding saturated sodium bicarbonate (50.0 mL), washed with saturated brine (50 mL*2), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was washed with ethanol (20.0 mL), filtered, and the dried filter cake was obtained as compound M-2. 1 H NMR (400 MHz, CDCl3) δ 10.62(s, 1H), 7.62 (d, J = 3.6 Hz, 1H),7.47 (d, J = 3.6 Hz, 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] + .

[0119] Third step

[0120] To a solution of compound M-2 (3.00 g, 10.37 mmol, 1 eq) in methanol (10.0 mL) was added sodium borohydride (980.7 mg) at 0 °C, and the reaction was stirred at 0 °C for 2 h. After the reaction was completed, saturated ammonium chloride solution (200.0 mL) was added to the reaction at 0 °C. Then it was diluted with water (100.0 mL), and the mixture was extracted with ethyl acetate (20.0 mL*2), washed with water (50.0 mL), saturated brine (50.0 mL), and the combined organic phase was 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 = 2.2: 1) to give compound M-C. 1 H NMR (400 MHz, CDCl3) δ 7.53 (d, J = 3.60 Hz, 1H), 6.74 (s, 1H), 6.61 (d,J = 3.60 Hz, 1H), 4.89 (s, 2H), 3.90 (s, 3H), 2.63 (s, 3H), 1.62 (s, 9H)。

[0121] Fourth step

[0122] To a solution of compound M-C (1.60 g) in methanol (1.2 mL) and dichloromethane (20.0 mL) was added chloromethylidene dimethylammonium chloride (1.41 g) at 20 °C, and the reaction was stirred for 1 h. The temperature was lowered to 0 °C, and saturated sodium bicarbonate solution (20.0 mL) was added to quench, and extracted with ethyl acetate (30 mL*3), and the combined organic phase was washed with saturated brine (20.0 mL*2) and dried over anhydrous sodium sulfate. It was filtered, and the filtrate was concentrated under reduced pressure to give compound M-1.

[0123] Example 1 Preparation of compounds 1A, 1B, 1C and 1D

[0124]

[0125]

[0126] First step

[0127] Compound 1-1 (3.20 g) was dissolved in tetrahydrofuran (30 mL), and the reaction solution was placed at -78 ℃ under nitrogen protection. Lithium bis(trimethylsilyl)amide (1 M, 10.5 mL) was added, and the reaction was incubated and stirred for 1 hour, and then warmed to 0 ℃. Compound 1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide (5.10 g) was added. The reaction solution was warmed to 30 ℃, and stirred for 12 hours. Water (15 mL) was added for quenching, and extracted with ethyl acetate (40 mL * 3). The organic phase was washed with saturated brine (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 ~ 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] + .

[0128] Second step

[0129] Compound 1-2 (0.50 g) and cyclobutylboronic acid (139.3 mg) were dissolved in toluene (10 mL) at 20 ℃, and 1,1-bis(diphenylphosphino)ferrocene palladium (II) dichloromethane complex (175.1 mg) and cesium carbonate (1.05 g) were added. The reaction solution was warmed to 110 ℃ and stirred for 14 hours under nitrogen atmosphere. The reaction was cooled to room temperature, and ice water (20 mL) was added for quenching, and extracted with ethyl acetate (25 mL*3). The organic phase was 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 : 10) to obtain compound 1-3. 1H NMR (400MHz, 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] + .

[0130] Third step

[0131] To the mixture 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, the reaction solution was heated to 100 ℃ and stirred for 20 hours. The reaction solution was cooled to room temperature, adjusted to pH < 7 with 50% potassium bisulfate aqueous solution, extracted with ethyl acetate (40 mL*3), the combined organic phase was washed with water (30 mL), saturated brine (25 mL) once, and dried over anhydrous sodium sulfate. Filtration, the filtrate was concentrated under reduced pressure to obtain compound 1-4. LC-MS: m / z = 392.1[M+H] + .

[0132] Fourth step

[0133] To the mixture solution of compound 1-4 (0.36 g) in methanol (1.2 mL) and toluene (3.6 mL), trimethylsilyldiazomethane (2 M, 919 µL) was added at 20 ℃, the reaction solution was stirred at 20 ℃ for 1 hour. The reaction solution was cooled to 0 ℃, quenched with acetic acid, diluted with water (40 mL), extracted with ethyl acetate (35 mL*3), the combined organic phase was washed with saturated sodium bicarbonate aqueous solution (25 mL), water (25 mL), saturated brine (25 mL) in turn, dried over anhydrous sodium sulfate, and filtered. 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 1-5. LC-MS: m / z = 406.1[M+H] + .

[0134] Fifth step

[0135] To a solution of compound 1-5 (0.24 g) in ethanol (4 mL) was added wet palladium on carbon (10%, 0.20 g) and hydrogen chloride-dioxane solution (4 M, 0.04 mL) at 20 °C. The reaction was stirred at 20 °C under hydrogen atmosphere (15 psi) for 1 h. The reaction was filtered, and the filtrate was concentrated under reduced pressure to give compound 1-6. LC-MS: m / z = 274.1 [M+H] + .

[0136] Step 6

[0137] To a solution of compound 1-6 (0.55 g) in N,N-dimethylformamide (15 mL) was added compound M-1 (1.01 g), cesium carbonate (1.64 g) and potassium iodide (334.0 mg) at 20 °C. The reaction was stirred at 20 °C for 15 h. The reaction was slowly poured into water (80 mL) and extracted with ethyl acetate (90 mL*3), the organic phase was combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by column chromatography on silica gel (ethyl acetate: petroleum ether = 1:1). The obtained crude product was further separated and purified by preparative high performance liquid chromatography (column: Phenomenex Gemini-NX 80*30 mm*3 µm; mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; gradient: acetonitrile%: 57%-87%). The obtained compound 1-7 was further separated by chiral separation (column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 µm; mobile phase: A: carbon dioxide, B: [0.1% ammonia water-isopropyl alcohol]; gradient: B%: 35%-35%). Compound 1-7A, compound 1-7B, compound 1-7C and compound 1-7D were obtained.

[0138] SFC analytical detection method: column: Chiralpak AD-3 150 mm*4.6 mm I.D., 3 µm, mobile phase: A: carbon dioxide, B: isopropyl alcohol (0.05% diethylamine), gradient: B%: 5% ~40% gradient flow for 5 minutes, 40% for 5 minutes, 5% for 2.5 minutes, flow rate: 2.5 mL / min. Compound 1-7A retention time: 4.238 minutes, ee: 100%; Compound 1-7B retention time: 4.946 minutes, ee: 100%; Compound 1-7C retention time: 5.530 minutes, ee: 99.4%; Compound 1-7D retention time: 5.954 min, ee: 100%.

[0139] Compound 1-7C The carbon hydrogen connected with piperidine ring and tetra ring butyl group has NOE correlation with the carbon hydrogen connected with benzoic acid, which is cis structure. Compound 1-7D The carbon hydrogen connected with piperidine ring and tetra ring butyl group has NOE correlation with the carbon hydrogen connected with benzoic acid, which is cis structure. LC-MS: m / z = 547.2 [M+H] + .

[0140] Seventh step

[0141] To the mixture solution of compound 1-7A (15.0 mg) in methanol (1 mL) and tetrahydrofuran (0.5 mL) was added lithium hydroxide monohydrate (3.5 mg), and the reaction solution was stirred at 50 ℃ for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography preparation separation (column: Phenomenex Gemini-NX 80*30 mm*3 µm; mobile phase: [water (10mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 16%-86%) to obtain compound 1A. LC-MS: m / z = 433.2 [M+H] + .

[0142] To the mixture solution of compound 1-7B (20.0 mg) in methanol (1 mL) and tetrahydrofuran (0.5 mL) was added lithium hydroxide monohydrate (4.6 mg), and the reaction solution was stirred at 50 ℃ for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography preparation separation (column: Phenomenex Gemini-NX 80*30 mm*3 µm; mobile phase: [water (10mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 16%-86%) to obtain compound 1B. LC-MS: m / z = 433.3 [M+H] + .

[0143] To the mixture solution of compound 1-7C (0.10 g) in methanol (1 mL) and tetrahydrofuran (0.5 mL) was added lithium hydroxide monohydrate (23.0 mg), and the reaction solution was stirred at 50 ℃ for 15 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography preparation separation (column: Phenomenex Gemini-NX 80*30 mm*3 µm; mobile phase: [water (10mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 16%-86%) to obtain compound 1C. 1H NMR (400 MHz, DMSO-d6) δ ppm 10.80 (s, 1H), 7.95 (d, J = 8.80 Hz, 2H), 7.64 (d, J = 7.60 Hz, 2H), 7.24(t, J = 2.80 Hz, 1H), 6.65 (s, 1H), 6.45 (t, J = 2.40 Hz, 1H), 3.70 (s, 3H),3.54 (d, J = 12.00 Hz, 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] + .

[0144] To a mixture of compound 1-7D (0.10 g) in methanol (1 mL) and tetrahydrofuran (0.5 mL) was added lithium hydroxide monohydrate (23.0 mg), and the reaction was stirred at 50 °C for 15 hours. The reaction was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini-NX 80*30 mm*3 µm; mobile phase: [water (10 mM ammonium bicarbonate) - acetonitrile]; acetonitrile%: 16%-86%) to give compound 1D. 1H NMR (400 MHz, DMSO-d6) δ ppm 10.80 (s, 1H), 7.95 (d, J = 8.40 Hz, 2H), 7.64 (d, J = 7.60 Hz, 2H), 7.24(t, J = 2.80 Hz, 1H), 6.65 (s, 1H), 6.43 - 6.47 (m, 1H), 3.70 (s, 3H), 3.54(d, J = 11.60 Hz, 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] + .

[0145] Preparation of compounds 2A, 2B, 2C and 2D of Example 2

[0146]

[0147]

[0148]

[0149] First step

[0150] A solution of titanium tetrachloride in dichloromethane (1 M, 1.2 mL) was dissolved in tetrahydrofuran (1 mL) at 0 ℃, a solution of compound 1-1 (0.20 g) and dimethyl maleate (79.0 mg) in tetrahydrofuran (1 mL) was added, the reaction solution was stirred at 0 ℃ for 1 hour, pyridine (236.6 mg) was added, the reaction solution was continued to stir at 20 ℃ for 15 hours. The reaction solution was poured into 10% citric acid to adjust pH < 7, extracted with ethyl acetate (20 mL*3), the combined organic phase was dried over anhydrous sodium sulfate, filtered, 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 = 8.40 Hz, 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] + .

[0151] Second step

[0152] To the solution of compound 2-2 (0.20 g) in methanol (2 mL) was added sodium borohydride (16.9 mg) at 0 ℃. The reaction solution was warmed to 25 ℃ and stirred for 3 hours. Ice water (30 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (30 mL*3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 2-3. LC-MS: m / z = 451.4 [M+H] + .

[0153] Third step

[0154] To the solution of compound 2-3 (1.10 g) in ethanol (15 mL) was added sodium borohydride (923.7 mg) at 25 ℃. The reaction solution was warmed to 70 ℃ and stirred for 2 hours. The reaction solution was cooled to room temperature, and ice water (50 mL) was added to quench the reaction. The mixture was extracted with ethyl acetate (50 mL), and the combined organic phase was 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 give compound 2-4. LC-MS: m / z = 395.1 [M+H] + .

[0155] Fourth step

[0156] To a solution of compound 2-4 (0.50 g) in tetrahydrofuran (6 mL) was added sodium hydride (60% purity, 60.8 mg) at 0 °C. The reaction was stirred at 0 °C for 0.5 h, then methylsulfonyl chloride (0.24 g) was added. The reaction was stirred at 25 °C for 1.5 h. The reaction was quenched by ice water (5 mL), extracted with ethyl acetate (10 mL*3), and the combined organic phase was dried over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by column chromatography on silica gel (ethyl acetate: petroleum ether = 2: 1) to give compound 2-5. LC-MS: m / z = 473.1 [M+H] + .

[0157] Step 5

[0158] To a solution of compound 2-5 (1.30 g) in tetrahydrofuran (40 mL) was added sodium hydride (60% purity, 330.1 mg) at 0 °C. The reaction was stirred at 20 °C for 1 h, then heated to 60 °C and stirred for 16 h. The reaction was quenched by ice water (60 mL), extracted with ethyl acetate (10 mL*3), and the combined organic phase was dried over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by column chromatography on silica gel (ethyl acetate: petroleum ether = 2: 1) to give compound 2-6. LC-MS: m / z = 377.1 [M+H] + .

[0159] Step 6

[0160] To a mixture of compound 2-6 (0.55 g) in isopropanol (2 mL), dioxane (2 mL) and water (4 mL) was added barium hydroxide (1.85 g) at 100 °C. The reaction was stirred at 100 °C for 14 h. The reaction was quenched by 50% potassium bisulfate aqueous solution to adjust pH < 7, extracted with ethyl acetate (40 mL*3), and the combined organic phase was washed with water (40 mL), saturated brine (45 mL) successively, and dried over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure to give compound 2-7 (0.58 g). LC-MS: m / z = 396.1 [M+H] + .

[0161] Step 7

[0162] To a solution of compound 2-7 (0.58 g) in methanol (1.5 mL) and toluene (4.5 mL) was added trimethylsilyldiazomethane (2 M, 1.47 mL) at 20 °C. The reaction solution was stirred at 20 °C for 2 h. The reaction solution was cooled to 0 °C, quenched with acetic acid, diluted with water (40 mL), extracted with ethyl acetate (45 mL*3), the combined organic phase was washed with saturated aqueous sodium bicarbonate solution (45 mL), water (45 mL), saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel (ethyl acetate: petroleum ether = 2: 1) to give compound 2-8. LC-MS: m / z = 410.1 [M+H] + .

[0163] Eighth step

[0164] To a solution of compound 2-8 (0.40 g) in methanol (5 mL) was added wet palladium on carbon (10% loading, 0.2 g) at 20 °C. The reaction solution was stirred at 20 °C under hydrogen atmosphere (15 psi) for 1 h. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to give compound 2-9. LC-MS: m / z = 276.1 [M+H] + .

[0165] Ninth step

[0166] To a solution of compound 2-9 (0.26 g) in N,N-dimethylformamide (6 mL) was added compound M-1 (0.65 g), cesium carbonate (769.2 mg), and potassium iodide (156.8 mg) at 20 °C. The reaction solution was stirred at 20 °C for 16 h. The reaction solution was slowly poured into water (50 mL), extracted with ethyl acetate (20 mL*3), the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (ethyl acetate: petroleum ether = 1: 2) and then by preparative high performance liquid chromatography (column: Phenomenex Gemini-NX 80*30 mm*3 µm; mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 57%-87%) to give compound 2-10. Chiral separation of compound 2-10 (column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 µm); mobile phase: Phase A: carbon dioxide, Phase B: [0.1% ammonia water-isopropyl alcohol]; B%: 40%-40%) gave compound 2-10A, compound 2-10B, compound 2-10C, and compound 2-10D.

[0167] SFC analytical detection method: Column: Chiralpak AD-3 150 mm*4.6 mm I.D., 3 µm, mobile phase: A: Carbon dioxide, B: Isopropanol (0.05% diethylamine), gradient: mobile phase B: 5% ~40% gradient flow for 5 minutes, 40% for 5 minutes, 5% for 2.5 minutes, flow rate: 2.5 mL / min. Compound 2-10A retention time: 4.333 minutes, 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] + .

[0168] Tenth step

[0169] To the mixture of compound 2-10A (80.0 mg) in methanol (3 mL) and tetrahydrofuran (1.5 mL) was added lithium hydroxide monohydrate (18.4 mg), the reaction solution was warmed to 50 °C and stirred for 14 hours. The reaction solution was concentrated under reduced pressure, the residue was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini-NX 80*30 mm*3 µm; mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 15%-45%) to give compound 2A. 1H NMR (400 MHz, CD3OD) δ ppm 8.12 (d, J = 8.0 Hz, 1H), 8.12 (d, J = 8.0 Hz, 1H), 7.61 (d, J = 8.0 Hz, 2H), 7.31 (d, J = 2.4 Hz, 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] + .

[0170] To the mixture of compound 2-10B (80.0 mg) in methanol (3 mL) and tetrahydrofuran (1.5 mL) was added lithium hydroxide monohydrate (18.4 mg), and the reaction solution was stirred at 50 °C for 14 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini-NX 80*30 mm*3 µm; mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile%: 15%-45%) to give compound 2B. Two-dimensional nuclear magnetic resonance identified that the hydrogen of piperidine connected to the four-membered ring oxygen was in a flat bond, and the hydrogen connected to benzoic acid was in an upright bond, and compound 2B was in a trans configuration. 1H NMR (400MHz, CD3OD) δ ppm 8.13 (d, J = 8.4 Hz, 2H), 7.61 (d, J = 8.4 Hz, 2H), 7.30(d, J = 3.2 Hz, 1H), 6.72 (s, 1H), 6.31(d, J = 2.8 Hz, 1H), 4.85-4.91 (m,2H), 4.36-4.46 (m, 3H), 4.27 (d, J = 12.8 Hz, 1H), 4.03 (d, J = 12.8 Hz, 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, 1H), 2.03-2.10 (m, 1H), 1.78-1.81 (m, 1H), 1.59-1.63(m, 1 H); LC-MS: m / z = 435.2 [M+H] + .

[0171] To the mixture of compound 2-10C (0.02 g) in methanol (1 mL) and tetrahydrofuran (0.5 mL) was added lithium hydroxide monohydrate (4.6 mg), and the reaction was stirred at 50 °C for 14 hours. The reaction was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini-NX 80*30mm*3µm; mobile phase: [water (10mM ammonium bicarbonate) - acetonitrile]; acetonitrile%: 15%-45%) to give compound 2C. The di-dimensional NMR identified that the hydrogen of piperidine and the hydrogen of the carbon connected with tetra-cyclic oxygen were both upright bond, and compound 2C was cis configuration. 1H NMR (400 MHz, CD3OD) δ ppm 8.13 (d, J = 8.0 Hz, 2H), 7.61 (d, J = 8.0 Hz, 2H), 7.31 (d, J = 3.2 Hz, 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, 1H), 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] + .

[0172] To the mixture of compound 2-10D (0.02 g) in methanol (1 mL) and tetrahydrofuran (0.5 mL) was added lithium hydroxide monohydrate (4.6 mg), and the reaction was stirred at 50 °C for 14 hours. The reaction was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini-NX 80*30 mm*3 µm; mobile phase: [water (10 mM ammonium bicarbonate) - acetonitrile]; acetonitrile%: 15%-45%) to give compound 2D. The 2D was confirmed by 2D NMR that the piperidine and the tetrahydrofuran were in cis configuration. 1H NMR (400 MHz, CD3OD) δ ppm 8.14 (d, J = 8.0 Hz, 2H), 7.62 (d, J = 8.0 Hz, 2H), 7.31 (d, J = 3.2 Hz, 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, 1 H), 2.51 (s, 3 H), 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] + .

[0173] Example 3 Preparation of compounds 3A, 3B, 3C and 3D

[0174]

[0175]

[0176] First step

[0177] 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 palladium dichloride dichloromethane complex (5.8 mg) were added, the reaction solution was stirred at 90 °C under nitrogen protection for 4.5 hours. The reaction solution was concentrated under reduced pressure, quenched with water (5 mL), extracted with ethyl acetate (10 mL), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated by filtration. The residue was separated and purified by silica gel thin layer chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain compound 3-3. 1H NMR (400 MHz, CDCl3) δ7.38-7.61 (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] + .

[0178] Second step

[0179] Compound 3-3 (0.3 g) was dissolved in a mixed solution of isopropyl alcohol, 1,4-dioxane and water (2 mL : 2 mL : 5 mL), barium hydroxide (573.6 mg) was added to the reaction solution, which was then heated to 100 °C and stirred for 12 hours. The reaction solution was cooled to room temperature, 50% potassium bisulfate aqueous solution was added to adjust pH = 5-6, 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] + .

[0180] Third step

[0181] Compound 3-4 (50.0 mg) was dissolved in a mixed solution of methanol (1.5 mL) and toluene (0.5 mL), (trimethylsilyl) diazomethane (2 M, 199 µL) was added at room temperature, and the reaction solution was stirred for 12 hours. Acetic acid (1 mL) and water (10 mL) were added for quenching, extracted with ethyl acetate (10 mL x 3), the organic phase was washed with saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by 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), 0.38-0.73 (m, 4H);LC-MS: m / z =392.2 [M+H] + .

[0182] Fourth Step

[0183] Compound 3-5 (250.0 mg) was dissolved in a mixed solution 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 under H2(15 Psi) for 2 hours. The reaction solution was directly filtered and concentrated under reduced pressure. 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] + .

[0184] Fifth Step

[0185] Compound 3-6 (0.20 g) was dissolved in 1,2-dichloroethane (5 mL), and compound M-2 (133.9 mg) and sodium triacetyl cyanoborohydride (269.7 mg) were added. The reaction solution was stirred at 15 °C for 20 hours. Water (3 mL) was added for quenching, and extraction was performed with ethyl acetate (10 mL x 3). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated and purified by high performance liquid chromatography (column type: 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 type: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 µm); mobile phase: A phase: carbon dioxide, B phase: [0.1% ammonia water-ethanol]; B%: 35%-35%) to obtain compound 3-7A, compound 3-7B, compound 3-7C, and compound 3-7D.

[0186] SFC detection method: Column: Chiralpak AD-3 150 mm*4.6 mm I.D., 3 µm, Mobile phase: A: Carbon dioxide, B: Isopropanol (0.05% diethylamine), Gradient: B%: 5% ~ 40% gradient flow for 5 minutes, 40% for 5 minutes, 5% for 2.5 minutes, Flow rate: 2.5 mL / min. Compound 3-7A retention time: 3.781 minutes, ee: 100%; Compound 3-7B retention time: 4.455 minutes, ee: 74.9%; Compound 3-7C retention time: 5.030 minutes, ee: 100%; Compound 3-7D retention time: 5.527 min, ee: 97.2%.

[0187] Step 6

[0188] Compound 3-7A (10.0 mg) was dissolved in a mixture of tetrahydrofuran (0.2 mL) and methanol (0.4 mL) at 20 °C, then lithium hydroxide aqueous solution (1 M, 0.2 mL) was added, and the reaction was stirred at 20 °C for 36 hours. 50% potassium hydrogen sulfate aqueous solution was added to adjust pH = 6-7, extracted with ethyl acetate (10 mL), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and 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]; acetonitrile%: 20%-60%), to obtain compound 3A. 1 H NMR (400MHz, CD3OD) δ 8.24 (d, J=8.2 Hz, 2H), 7.76 (d, J=8.2 Hz, 2H), 7.35 (d, J=3.2Hz, 1H), 6.78 (s, 1H), 6.39 (d, J=3.0 Hz, 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] + .

[0189] Compound 3-7B (20.0 mg) was dissolved in a mixture of tetrahydrofuran (0.3 mL) and methanol (0.6 mL) at 20 °C, then lithium hydroxide aqueous solution (1 M, 0.3 mL) was added, the reaction was stirred at 20 °C for 36 hours. 50% potassium hydrogen sulfate aqueous solution was added, pH = 6-7 was adjusted, the mixture was concentrated under reduced pressure, 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]; acetonitrile%: 20%-60%), to obtain compound 3B. 1 H NMR (400 MHz, CD3OD) δ 8.24 (d, J = 8.2 Hz, 2H), 7.76(d, J = 8.2 Hz, 2H), 7.35 (d, J = 3.0 Hz, 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] + .

[0190] Compound 3-7C (18.0 mg) was dissolved in a mixture of tetrahydrofuran (0.3 mL) and methanol (0.6 mL) at 20 °C, then lithium hydroxide monohydrate (1 M, 0.3 mL) was added, the reaction was stirred at 20 °C for 36 hours. 50% potassium hydrogen sulfate aqueous solution was added, pH = 6-7 was adjusted, the mixture was concentrated under reduced pressure, 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]; acetonitrile%: 20%-60%), to obtain compound 3C. 1H NMR (400 MHz, CD3OD) δ 8.23 (d, J = 8.0 Hz, 2H), 7.67-7.80 (m, 2H), 7.33 (d, J = 3.0 Hz, 1H), 6.77 (s, 1H), 6.34 (d, J = 3.0 Hz, 1H), 4.45-4.55 (m, 1H), 4.34 (d, J = 12.4 Hz, 1H), 4.13 (d, J = 12.4 Hz, 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] + .

[0191] Compound 3-7D (20.0 mg) was dissolved in a mixture of tetrahydrofuran (0.3 mL) and methanol (0.6 mL) at 20 °C, then lithium hydroxide monohydrate (1 M, 0.3 mL) was added, and the reaction was stirred at 20 °C for 36 hours. 50% potassium bisulfate aqueous solution was added to adjust pH = 6-7, and the mixture was concentrated under reduced pressure. 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]; acetonitrile%: 20%-60%), to obtain compound 3D. 1H 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] + .

[0192] Preparation of compounds 4A and 4B of Example 4

[0193]

[0194]

[0195]

[0196] First step

[0197] Potassium tert-butoxide (1.40 g) was added to a solution of trimethylsulfoxonium iodide (2.50 g) in 1,2-dichloroethane (15.0 mL) at 25 °C under nitrogen protection. The reaction solution was stirred for 0.5 h, the temperature was lowered to 0 °C, and a solution of compound 1-1 (3.50 g) in 1,2-dichloroethane (15.0 mL) was added. The reaction was warmed to 25 °C and stirred for 12 h. Water (10.0 mL) was added for quenching, and extraction was performed with ethyl acetate (50.0 mL*3). The organic phase was washed with saturated brine, 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%~35%) to obtain compound 4-2. LC-MS: m / z =349.1 [M+H] + .

[0198] Second step

[0199] Compound 4-2 (1.70 g) was dissolved in dichloromethane (20.0 mL), then boron trifluoride etherate (1.04 g) was added, and the reaction was stirred at 25 °C for 12 hours. Water (5.0 mL) was added for quenching, and extraction was performed with dichloromethane (20.0 mL * 3), and the organic phase was washed with saturated brine (20.0 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 ~ 2 : 1) to obtain compound 4-3. LC-MS: m / z = 349.1 [M+H] + .

[0200] Third step

[0201] Sodium tert-butoxide (744.8 mg) was added to a solution of methyltriphenylphosphonium iodide (3.13 g,) in tetrahydrofuran (15.0 mL) at 25 °C under nitrogen protection, and the reaction was stirred at 25 °C for 1 hour. Then a solution of compound 4-3 (1.80 g) in tetrahydrofuran (10.0 mL) was added, and the reaction was continued to stir for 12 hours. Water (10.0 mL) was added for quenching, and extraction was performed with ethyl acetate (20 mL * 3), and the organic phase was 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 ~ 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 (brs, 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] + .

[0202] Fourth step

[0203] Compound 4-4 (1.20 g), zinc copper reagent (4.47 g) and phosphorus oxychloride (584.3 mg) were dissolved in ethylene glycol dimethyl ether (50.0 mL) at 20 °C under nitrogen protection, and trichloroacetyl chloride (3.15 g) was added. The reaction solution was stirred at 20 °C for 12 hours. It was concentrated under reduced pressure, and the residue was separated and purified by flash column chromatography on silica gel (ethyl acetate / petroleum ether, ethyl acetate ratio 0-50%) to obtain compound 4-5. LC-MS: m / z = 457, 459 [M+H] + .

[0204] Fifth step

[0205] 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 the solution was stirred for 1 hour. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by flash column chromatography on silica gel (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=8.4 Hz, 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).

[0206] Sixth step

[0207] Compound 4-6 (1.2 g) was dissolved in dichloromethane (30.0 mL) at 15 °C, the temperature was lowered to -15 °C, diethylaminosulfur trifluoride (2.49 g) was added, the temperature was slowly raised to 15 °C, and the stirring was continued for 12 hours. The reaction solution was slowly poured into ice water (100.0 mL) to quench, the pH was adjusted to 7 with saturated aqueous sodium bicarbonate solution, and it was extracted with dichloromethane (100 mL*3), the organic phases were combined, and concentrated under reduced pressure. The residue was separated and purified by flash column chromatography on silica gel (ethyl acetate / petroleum ether, ethyl acetate ratio 0-50%) to obtain compound 4-7. 1H NMR (400 MHz, CDCl3) δ 7.64 (d, J=8.4 Hz, 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).

[0208] Seventh step

[0209] 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) at 20 °C, and barium hydroxide (960.1 mg) was added. The temperature was raised to 100 °C, and the reaction was stirred for 12 h. The reaction was cooled to room temperature, poured into water (100.0 mL) and quenched. The pH was adjusted to 5-6 with 1 M hydrochloric acid, and the mixture was extracted with ethyl acetate (100 mL*3). The organic phase was combined and concentrated under reduced pressure to give compound 4-8. LC-MS: m / z = 430 [M+H] + .

[0210] Eighth step

[0211] Compound 4-8 (0.98 g) was dissolved in a mixture of methanol (5.0 mL) and toluene (15.0 mL) at 20 °C, and then trimethylsilyldiazomethane (2 M, 2.45 mL) was slowly added. The reaction was stirred at 20 °C for 0.5 h. Acetic acid (2.0 mL) was slowly added to quench the reaction, and then the mixture was concentrated under reduced pressure. The concentrate was purified by flash silica gel column chromatography (ethyl acetate / petroleum ether, ethyl acetate ratio 0-50%) to give compound 4-9. Two-dimensional nuclear magnetic resonance (HSQC, COSY, NOE) analysis showed that the C-H of the piperidine connected to the difluorocyclobutane was a straight bond, and had NOE correlation with the two hydrogens in the alpha position of the benzene ring, so it was determined that the relative configuration of the difluorocyclobutane and the methyl benzoate was the trans configuration. 1H NMR (400 MHz, CDCl3) δ 8.04 (br d, J=8.4 Hz,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] + .

[0212] Ninth step

[0213] Compound 4-9 (900.0 mg) was dissolved in ethyl acetate (15.0 mL) at 25 °C, then palladium hydroxide (180.0 mg) in methanol (5.0 mL) was added, the system was replaced with hydrogen gas for 3 times, then the hydrogen pressure was maintained at 15 psi, and the system was stirred at room temperature for 2 hours. Filtration and concentration of the filtrate under reduced pressure gave compound 4-10. 1 H NMR (400 MHz, CDCl3) δ 8.00 (d, J=8.4 Hz, 2H), 7.45 (d, J=8.4 Hz, 2H), 3.92-3.98 (m, 1H), 3.91 (s, 3H), 2.953-2.96 (m, 2H), 2.64-2.76 (m, 2H), 2.38-2.52 (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] + .

[0214] Tenth step

[0215] Compound 4-10 (70.0 mg) and compound M-1 (180.0 mg) were dissolved in N,N-dimethylformamide (3.0 mL) at 25 °C, then cesium carbonate (221.2 mg) and potassium iodide (37.6 mg, 226.28 μmol, 1 eq.) were added, and the reaction was incubated and stirred for 12 hours. The reaction was quenched by pouring into water (100 mL), extracted with ethyl acetate (70 mL*3), the organic phases were combined and concentrated under reduced pressure. The residue was purified by silica gel thin layer chromatography (petroleum ether: ethyl acetate = 3: 1) to obtain compound 4-11, which was further separated by chiral separation (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 compound 4-11A and compound 4-11B.

[0216] SFC analysis detection method: Column: Cellulose 2 150 mm*4.6 mm I.D., 5 μm, mobile phase: A: carbon dioxide, B: ethanol (0.05% diethylamine), gradient: B%: 5% ~40% gradient flow for 5 minutes, 40% for 5 minutes, 5% for 2.5 minutes, flow rate: 2.5 mL / min. Compound 4-11A retention time: 3.267 min, compound 4-11B retention time: 3.514 min. LC-MS: m / z =583 [M+H] + .

[0217] Tenth step

[0218] Compound 4A:

[0219] Compound 4-11A (50.0 mg) was dissolved in a mixture solution of tetrahydrofuran (1.0 mL), methanol (2.0 mL) and water (1.0 mL) at 20 °C, then lithium hydroxide (40.0 mg) was added, the reaction solution was warmed 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 solution, extracted with ethyl acetate (30 mL*3), the organic phase was collected, washed with saturated brine (20.0 mL), dried over anhydrous sodium sulfate, filtered, concentrated, the residue was dissolved in a mixture solvent of water (10.0 mL) and acetonitrile (2.0 mL), and freeze-dried to obtain compound 4A (SFC detection method: column: Chiralpak AD-3 150 mm*4.6 mm I.D., 3 μm, mobile phase: A: carbon dioxide, B: isopropyl alcohol (0.05% diethylamine), gradient: B%: 5% ~ 40% gradient flow for 5.5 minutes, 40% for 3 minutes, 5% for 1.5 minutes, flow rate: 2.5 mL / min, retention time: 6.521 minutes, ee: 99.7%). 1 H NMR (400 MHz, DMSO-d6) δ: 10.80(br s, 1H), 7.89 (d, J=8.4 Hz, 2H), 7.52 (br d, J=8.0 Hz, 2H), 7.24 (t, J=2.8Hz, 1H), 6.64 (s, 1H), 6.48 (t, J=2.4 Hz, 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] + .

[0220] Compound 4B:

[0221] Compound 4-11B (46.0 mg) was dissolved in a mixture solution of tetrahydrofuran (1.0 mL), methanol (2.0 mL) and water (1.0 mL) at 20 °C, then lithium hydroxide (40.0 mg) was added, the reaction solution was warmed 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 solution, extracted with ethyl acetate (30 mL*3), the organic phase was collected, washed with saturated brine (20.0 mL), dried over anhydrous sodium sulfate, filtered, concentrated, the residue was dissolved in a mixture solvent of water (10.0 mL) and acetonitrile (2.0 mL), and freeze-dried to obtain compound 4B (SFC detection method: column: Chiralpak AD-3 150 mm*4.6 mm I.D., 3 μm, mobile phase: A: carbon dioxide, B: isopropyl alcohol (0.05% diethylamine), gradient: B%: 5% ~ 40% gradient flow for 5.5 minutes, 40% for 3 minutes, 5% for 1.5 minutes, flow rate: 2.5 mL / min, retention time: 4.832 minutes, ee: 96.0%). 1 H NMR (400 MHz, DMSO-d6) δ:10.80(br s, 1H), 7.93 (d, J=8.0 Hz, 2H), 7.61 (br d, J=7.4 Hz, 2H), 7.24 (t, J=2.8Hz, 1H), 6.64 (s, 1H), 6.48 (t, J=2.0 Hz, 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] + .

[0222] Example 5 Preparation of compound 4C

[0223]

[0224]

[0225]

[0226] First step

[0227] 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 nitrogen protection and stirred for 16 hours. The reaction solution was poured into ice water (15.0 L), filtered, and the filter cake was washed with 2-methyltetrahydrofuran (2.0 L*3). The filtrate was collected, washed with saturated brine (3.0 L), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove 2-methyltetrahydrofuran (8.0 L). The remaining solution was poured into petroleum ether (10.0 L) at 0°C, stirred for 1 hour, filtered, and the filter cake was washed with petroleum ether (1.0 L*3) and 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).

[0228] Second step

[0229] To a solution of compound 5-3 (5.00 g) in 2-methyltetrahydrofuran (100.0 mL) was added dropwise lithium diisopropylamide (2 M tetrahydrofuran solution, 12.08 mL) at -20°C under nitrogen protection. After the dropwise addition was completed, the reaction solution was stirred at -20°C to -10°C for 0.5 hours. A solution of compound 5-4 (5.74 g) in 2-methyltetrahydrofuran (25.0 mL) was added, and the temperature was slowly increased. The reaction solution was stirred at -20°C to -10°C for 7 hours. The reaction was quenched by adding saturated aqueous ammonium chloride solution (30.00 mL), diluted with water (200.0 mL), and extracted with ethyl acetate (100.0 mL*2). The organic phases were combined, washed with saturated brine (100.0 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was 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

[0230] Third step

[0231] 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 was stirred at 0 °C for 3 h. Water (30.0 mL) was added to quench the reaction, and ethyl acetate (30.0 mL * 2) was used to extract the product. The organic phase was washed with saturated brine (30.0 mL * 1), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate, ethyl acetate ratio: 0-100%) to obtain compound 5-6. LC-MS: m / z = 399.2 [M+H] + .

[0232] Fourth step

[0233] Compound 5-6 (600.0 mg) was dissolved in tetrahydrofuran (15.0 mL) at 25 °C under nitrogen protection, triphenylphosphine (1.18 g) and diisopropyl azodicarboxylate (913.3 mg) were added. The reaction was stirred at 25 °C for 18 h. The reaction was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 0-50%) to obtain compound 5-7. LC-MS: m / z = 381.1 [M+H] + .

[0234] Fifth step

[0235] Compound 5-7 (200.0 mg) was dissolved in methanol (1.0 mL) at 20 °C under nitrogen protection, and concentrated H2SO4 (1.0 mL) was added. The reaction was heated to 80 °C and stirred for 3 h. The reaction was cooled to room temperature, diluted with water (50.0 mL), and adjusted to pH 7-8 with saturated sodium bicarbonate solution. The product was extracted with ethyl acetate (50.0 mL x 2). The combined organic phase was washed with saturated brine (50.0 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography on silica gel (dichloromethane: methanol, methanol ratio: 0-10%) to obtain compound 5-8. LC-MS: m / z = 310.2 [M+H] + .

[0236] Sixth step

[0237] To compound 5-8 (50.0 mg,) and compound M-2 (51.4 mg) in 1,2-dichloroethane (1.0 mL) was added sodium borohydride in acetic acid (137.0 mg) at 25 °C. The mixture was stirred at 25 °C for 16 hours. Water (200.0 mL) was added to dilute, extracted with ethyl acetate (50.0 mL * 2), combined organic phase was washed with water (100.0 mL * 2), dried over anhydrous sodium sulfate, filtered. The filtrate was concentrated under reduced pressure, 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] + .

[0238] Seventh step

[0239] Compound 5-9 (30.0 mg) was dissolved in tetrahydrofuran (0.5 mL) and methanol (0.5 mL) at 25 °C, and lithium hydroxide monohydrate (13.0 mg) was added. The reaction solution was stirred at 50 °C for 16 hours. The reaction solution was cooled to room temperature, acetic acid (0.1 mL) was added to quench, water (50.0 mL) was added to dilute, extracted with ethyl acetate (25.0 mL * 2), combined organic phase was dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure to obtain compound 4C.

[0240] LC-MS: m / z = 469.1 [M+H] + .

[0241] Preparation of compound 4D in Example 6

[0242]

[0243]

[0244]

[0245] First step

[0246] Compound 5-1 (50.00 g) and compound 6-2 (45.92 g) were dissolved in 2-methyltetrahydrofuran (500.0 mL), and tetraethoxytitanium (235.72 g) was added in batches at 25 °C. The reaction solution was heated to 80 °C under nitrogen protection and stirred for 16 hours. The reaction solution was poured into ice water (15.0 L), filtered, the filter cake was washed with 2-methyltetrahydrofuran (3.0 L), the filtrate was collected, dried over anhydrous sodium sulfate, filtered, and 2-methyltetrahydrofuran (400.0 mL) was removed under reduced pressure. The remaining solution was poured into petroleum ether (200.0 mL) at 0 °C and stirred for 1 hour, filtered, and the filter cake was collected and dried under reduced pressure to obtain compound 6-3. 1H NMR (400 MHz, CDCl3) δ: 7.95-7.97 (d, J=8.0 Hz, 2H), 7.72-7.74 (d, J=8.0 Hz, 2H), 2.79 (s, 3H), 1.33 (s, 9H).

[0247] Second step

[0248] To a solution of compound 6-3 (25.00 g) in 2-methyltetrahydrofuran (500.0 mL) was added dropwise lithium diisopropylamide (2M tetrahydrofuran solution, 60.40 mL) at -20 °C under nitrogen protection. After the dropwise addition was completed, the reaction solution was stirred at -20 °C to -15 °C for 1 hour. A solution of compound 5-4 (35.35 g) in 2-methyltetrahydrofuran (125.0 mL) was added, and the temperature was slowly increased. The reaction solution was stirred at -20 °C to -15 °C for 4 hours. The reaction was quenched by adding saturated aqueous ammonium chloride solution (60.00 mL), and diluted with water (300.0 mL). The organic phase was extracted with ethyl acetate (200.0 mL * 2), washed with water (200.0 mL * 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: 0-13%) to obtain compound 6-5.

[0249] Third step

[0250] Compound 6-5 (5.20 g) was dissolved in tetrahydrofuran (52.0 mL) at 0 °C, and sodium borohydride (0.50 g) was added. The reaction was stirred at 0 °C for 1 hour. The reaction was quenched by adding saturated ammonium chloride solution (50.0 mL), and diluted with water (200.0 mL). The organic phase was extracted with ethyl acetate (100.0 mL * 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] + .

[0251] Fourth step

[0252] Compound 6-6 (3.30 g) was dissolved in tetrahydrofuran (15.0 mL) at -15 °C, and lithium tri-sec-butylborohydride (1 M tetrahydrofuran solution, 5.02 mL) was added. The reaction was stirred at -15 °C for 4 hours. Saturated ammonium chloride solution (50.0 mL) was added to quench, and water (200.0 mL) was added to dilute. The mixture was extracted with ethyl acetate (50.0 mL * 3), and the organic phase was combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel (dichloromethane:methanol, methanol ratio: 0~5%) to obtain compound 6-7. LC-MS: m / z = 399.2 [M+H] + .

[0253] Fifth step

[0254] Compound 6-7 (2.30 g) was dissolved in tetrahydrofuran (46.0 mL) at 25 °C, and triphenylphosphine (5.30 g) and diisopropyl azodicarboxylate (4.08 g) were added. The reaction was stirred at 25 °C under nitrogen protection for 16 hours. The reaction was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate, ethyl acetate ratio: 0~40%) to obtain compound 6-8. LC-MS: m / z = 381.1 [M+H] + .

[0255] Sixth step

[0256] Compound 6-8 (500.0 mg) was dissolved in methanol (5.0 mL) at 20 °C under nitrogen protection, and concentrated H2SO4 (5.0 mL) was added. The reaction was heated to 80 °C and stirred for 16 hours. The reaction was cooled to room temperature, and the pH was adjusted to 7~8 with saturated aqueous sodium bicarbonate solution. The mixture was extracted with ethyl acetate (50.0 mL * 2). The organic phase was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by thin layer chromatography on silica gel (dichloromethane:methanol = 0~10%) to obtain compound 6-9. LC-MS: m / z = 310.1 [M+H] + .

[0257] Seventh step

[0258] To compound 6-9 (130.0 mg,) and compound M-2 (133.7 mg) in 1,2-dichloroethane (3.0 mL) was added sodium borohydride acetic acid (356.3 mg) at 25 °C. The mixture was stirred at 25 °C for 16 hours. Diluted with water (200.0 mL), extracted with ethyl acetate (50.0 mL * 2), the combined organic phase was washed with water (100.0 mL * 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure 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] + .

[0259] Eighth step

[0260] Compound 6-10 (30.0 mg) was dissolved in tetrahydrofuran (0.5 mL) and methanol (0.5 mL) at 25 °C, and lithium hydroxide monohydrate (13.0 mg) was added. The reaction solution was stirred at 50 °C for 16 hours. The reaction solution was cooled to room temperature, acetic acid (0.1 mL) was added to quench, diluted with water (50.0 mL), extracted with ethyl acetate (25.0 mL * 2), the combined organic phase was dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure to obtain compound 4D. LC-MS: m / z = 469.1 [M+H] + .

[0261] Biological test data:

[0262] Experimental Example 1 Wieslab Complement Alternative Pathway Activation Inhibition (enzyme activity test)

[0263] The purpose of this experiment is to determine the inhibitory activity of the compounds of the present application against the complement alternative pathway in human serum by Wieslab® Complement System Alternative Pathway Kit.

[0264] Experimental protocol:

[0265] Dilute the serum with diluent (1:23). Add drug to the diluted serum, 8 concentration gradients, the highest 10 mM or 50 mM, 5-fold gradient dilution. Incubate at room temperature for 15 min. Add the compound and serum mixture to the 96-well plate provided by the kit (100 µL / well), activate at 37 degrees for 1 hour. Wash three times with washing buffer. Add the detection antibody provided by the kit (100 µL) and incubate at room temperature for 30 min. Wash three times with washing buffer. Add the substrate (100 µL) and incubate at room temperature for 30 min. Detect the absorbance at 405 nM by microplate reader.

[0266] Experimental results: The results of the test compound on the complement bypass pathway inhibition activity are shown in Table 1.

[0267] Table 1: Results of in vitro enzyme activity screening test

[0268]

[0269] Note: *average of two tests

[0270] Conclusion: The compound of the present application has obvious inhibition activity on the activation of human serum bypass pathway.

[0271] Experimental Example 2 Mouse pharmacokinetic study test

[0272] Experimental purpose: To investigate the plasma pharmacokinetics of the compound of the present application in male C57BL / 6J mice after single intravenous injection and oral administration.

[0273] Experimental animals: Male C57BL / 6J mice, 7-9 weeks old, weighing 17-23 grams; supplier: Shanghai Xipu-Bike Experimental Animal Co., Ltd.

[0274] Experimental process: Injection administration (IV): the dose is 1 mg / kg (solvent: 10%PG / 5%Solutol / 85%PBS (1x pH 7.4)); oral administration (PO): the dose is 10 mg / kg (solvent: 30%PEG300 / 10%Cremphor EL / 60%PBS (1x pH 7.4))

[0275] Sample collection: 0.03 mL of blood sample was collected from the experimental animals at each time point by saphenous vein puncture, and the actual blood collection time was recorded. All blood samples were added to a commercial EDTA-K2 anticoagulation tube with a specification of 1.5 mL (supplier: Jiangsu Kangjian Medical Supplies Co., Ltd.). After blood sample collection, within half an hour, the supernatant plasma was centrifuged at 4°C, 3000 g for 10 minutes, quickly placed on dry ice, and stored in a -80°C refrigerator for LC-MS / MS analysis.

[0276] Data analysis: The plasma concentration was processed by non-compartment model using WinNonlin™ Version 6.3 (Pharsight, Mountain View, CA) pharmacokinetic software, and the pharmacokinetic parameters Cl , T 1 / 2 ,C max , AUC 0-last , the results are shown in Table 2.

[0277] Table 2 Comparison of PK results of the compound of the present application and the reference compound

[0278]

[0279] Note: Cl: apparent clearance; T 1 / 2 : time required for half of the compound to be cleared; C max : peak concentration; AUC 0-last : concentration integral area within 0- the last sampling time.

[0280] Experimental conclusion: The pharmacokinetic study results of the compound of the present application in mice show that the half-life is longer than that of the reference compound, and the oral plasma exposure is significantly better than that of the reference compound LNP023, and the pharmacokinetic properties of the compound of the present application are excellent.

[0281] Experimental Example 3: Rat pharmacokinetic study test

[0282] Experimental purpose: To investigate the plasma pharmacokinetics of the compound of the present application in male Wistar Han rats after single intravenous injection and oral administration.

[0283] Experimental animals: male Wistar Han rats, 7-9 weeks old, weighing 231.03-243.2 grams; supplier: Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0284] Experimental process: injection administration (IV): the dose is 1 mpk (solvent: 30% PEG300 / 10% Cremphor EL / 60% PBS (1x pH 7.4)); oral administration (PO): the dose is 5 mpk or 30 mpk (solvent: 30% PEG300 / 10% Cremphor EL / 60% PBS (1x pH 7.4))

[0285] Sample collection: blood samples of about 0.3 mL were collected from the jugular vein of experimental animals at each time point, and the actual blood collection time was recorded. All blood samples were added to commercial EDTA-K2 anticoagulation tubes with a specification of 1.5 mL (supplier: Jiangsu Kangjian Medical Supplies Co., Ltd.). After blood sample collection, centrifuge the supernatant plasma at 4℃, 3200 g for 10 minutes, quickly put on dry ice, and store in a-80℃ refrigerator for LC-MS / MS analysis.

[0286] Data analysis: the plasma concentration was processed by non-compartment model using Phoenix WinNonlin 6.3 pharmacokinetic software, and the pharmacokinetic parameters Cl, C max , T 1 / 2 , AUC 0-last were calculated by linear logarithmic trapezoidal method. The results are shown in Table 3.

[0287] Table 3 Comparison of rat PK results of the compound of the present application and the reference compound

[0288]

[0289] Note: Cl: apparent clearance; T 1 / 2 : time required for half of the compound to be cleared; C max : peak concentration (dose-normalized data in table); AUC 0-last : area under the concentration-time curve from 0 to the last sampling time (dose-normalized data in table); a: dose administered was 5 mpk; b: dose administered was 30 mpk.

[0290] Experimental conclusion: The pharmacokinetic research results of the compound of the present application in rats show that the half-life is longer than that of the reference compound, and the oral plasma exposure is significantly better than that of the reference compound LNP023, and the pharmacokinetic property of the compound of the present application is excellent.

[0291] Experimental Example 4: LPS-induced complement activation in vivo PD model in mice

[0292] Experimental purpose: To investigate the inhibitory effect of the compound of the present application on LPS-induced complement activation in mice.

[0293] Experimental animals: female C57BL / 6J mice, 7-9 weeks old, weighing 17-23 grams; supplier: Shanghai Xipu-Bike Experimental Animal Co., Ltd.

[0294] Experimental process: 100 μg of lipopolysaccharide (LPS) in Salmonella typhimurium (Sigma) was dissolved in 100 μL of sterile PBS to induce complement activation in mice by intraperitoneal injection. The negative control animals received intraperitoneal injection of 100 μL of sterile PBS and were administered by gavage (PO) alone. The positive control animals received intraperitoneal LPS and PO drug vehicle (0.5% (w / v) methylcellulose and 0.5% (v / v) Tween 80). The administration and plasma collection time points are shown in Table 2. Figure 1

[0295] Sample collection: blood samples of 0.3 mL were collected from the orbital plexus. All blood samples were added to commercial EDTA-K2 anticoagulant tubes with a specification of 1.5 mL (supplied by Jiangsu Kangjian Medical Supplies Co., Ltd.). After blood sampling, within half an hour, the supernatant plasma was centrifuged at 4℃, 3000 g for 10 minutes, quickly placed on dry ice, and stored in a -80℃ refrigerator for Western blot analysis of the downstream C3d protein level after complement activation.

[0296] ​Sample analysis: Mix mouse plasma (5 μL) + Lysis buffer (27.5 μL) + Loading buffer (12.5 μL) + Reducing buffer (5 μL) uniformly, incubate at 100 ℃ for 15 min, and load 5 μL per well, that is, 0.5 μL of plasma per well.

[0297] Experimental results: In this experiment, the Factor B inhibitor LNP023 of Novartis was used as a reference drug, and the test compounds were the reference compound LNP023, compound 3A and compound 3C. All three drugs can significantly inhibit the C3d level, that is, can inhibit the LPS-induced complement activation, and the inhibition effect of compound 3A and compound 3C is better than that of LNP023. The specific experimental results are shown in the following table, and the data are expressed as: Mean ± SEM, n = 5; ###, p < 0.001, normal group vs. model group, t-test; ****, p < 0.0001, drug group vs. model group, one-way ANOVA. Figure 2

[0298] Experimental conclusion: The compound of the present application can significantly inhibit the complement activation induced by LPS, and the inhibition effect is better than that of LNP023.

[0299] Experimental example 5: In vivo pharmacodynamic model of passive Heymann nephritis rats

[0300] Experimental purpose: To investigate the ability of the compound of the present application to improve the renal function of rats with Sheep Anti-Rat Fx1A Serum-induced Heymann nephritis, including reducing the proteinuria level and improving the renal tissue damage.

[0301] Experimental animals: Male SD rats, 7-10 weeks old, weighing 200-300 grams; supplier: Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0302] Experimental process:

[0303] 1. Modeling:

[0304] D-2 days before administration, collect rat urine. D1 is the first day of the experiment, and the control group (group 1) animals are given a single injection of 5 mL / kg of Sheep Non-Immune serum through the tail vein; the modeling group and the administration group (groups 2-6) animals are given a single injection of 5 mL / kg of Sheep Anti-Rat Fx1A Serum through the tail vein.

[0305] 2. Drug administration: ​

[0306] The administration mode is gavage, twice a day, with an 8-hour interval between administrations, and the administration volume is 10 mL / kg. On D1, 1 hour before modeling, the animals in groups 1 and 2 are given blank solvent 20% PEG 400 / 10% Solutol / 70% water, the animals in group 3 are given LNP023 (60 mpk), and the animals in groups 4-6 are given different concentrations of compound 4B (5 mpk, 20 mpk, and 60 mpk). After an 8-hour interval from the first administration, each group is administered again at the same dose and volume as the first administration. On D2-D14, the animals in each group are continuously administered different compounds or solvent for 14 days (including day 1) at the same dose, volume, administration mode, and frequency as on D1.

[0307] 3. Sample collection:

[0308] Urine collection: On D-2 days before administration, and on D4, D6, D8, D11, and D14 days, 2-4 h and 4-6 h after the first administration, urine samples from rats are collected into EP tubes and stored in a refrigerator at -80°C to -60°C for rat urine protein and urine creatinine detection.

[0309] Kidney sample collection: All animals are euthanized by CO2 inhalation anesthesia on Day 15, and bilateral kidneys are collected. The left kidney is cut transversely, and the right kidney is cut longitudinally. The transversely cut half (left) and the longitudinally cut half (right) are placed in formalin fixation (in the same EP tube).

[0310] 4. Sample analysis:

[0311] (1) Rat urine creatinine uCRE is normally loaded after ten-fold dilution (Decrease mode), 10 microliters of sample + 90 microliters of saline (0.9% saline), and more than the detection upper limit is diluted 20-fold (Increase mode), 5 microliters of sample + 95 microliters of saline (0.9% saline). Rat urine total protein uTP is detected at the original concentration, and more than the detection upper limit is diluted 10-fold (Decrease mode), 10 microliters of sample + 90 microliters of saline (0.9% saline), and if it is still more than the detection upper limit after ten-fold dilution, it is diluted 100-fold (Increase mode), 2 microliters of sample + 198 microliters of saline (0.9% saline). The data are read by the analysis instrument HITACHI LST008 AS(P).

[0312] (2) Kidney pathological score: The damage degree of the drug on the kidney (paraffin section) of the rat was analyzed by HE staining. The scoring criteria were as follows: 0 represented normal, 1 represented a little cell infiltration in the mesangium, 2 represented more cell infiltration in the mesangium, 3 represented several glomerular mesangial cell proliferation, several mesangial cell infiltration, 4 represented renal tubular cast, atrophy, glomerular crescent formation and sclerosis.

[0313] Experimental results: The urine protein and urine creatinine of the rat were detected, and the Factor B inhibitor LNP023 of Novartis was used as a reference drug. Both LNP023 and compound 4B could significantly inhibit the urine protein level of the rat and improve the kidney function of the rat, and the drug efficacy of compound 4B was better than that of LNP023 at a dose of 60 mpk. The specific experimental results are shown in the following table 3: Figure 3 , the data are represented as: Mean ± SEM, n = 5; ###, p < 0.001, normal group vs model group, t-test; ****, p < 0.0001, drug administration group vs model group, one-way ANOVA.

[0314] The pathological score results are shown in table 4:

[0315] Table 4 Pathological score results

[0316]

[0317] Experimental conclusion: According to the pathological score results, compared with the model group, the compounds of the present application in different dose groups could obviously improve the pathological degree of the kidney of the model animal. The compounds of the present application could reduce the urine protein level of the rat and improve the kidney function, and the drug efficacy was better than that of LNP023.

Claims

1. The compound represented by formula (I) or a pharmaceutically acceptable salt thereof, in, L is selected from single bonds, NR4, and O; R1 is selected from fluorine, chlorine, and 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-3-6-membered heterocyclic alkyl, wherein the 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-3-6-membered heterocyclic alkyl groups are each independently and optionally surrounded by 1, 2, or 3 R groups. a replace; R2, R3, and R4 are independently selected from H and C, respectively. 1-5 Alkyl, the C 1-5 Alkyl groups may be optionally surrounded by 1, 2, or 3 R's. b replace; Each R a and R b They are each independently selected from H, F, Cl, Br, and I; The condition is that R1 is selected from unsubstituted C. 1-5 When alkyl, R2 and R3 are not both selected from H.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from, in, L, R1, R2, and R3 are as defined in claim 1; Carbon atoms marked with "*" are chiral carbon atoms, existing as a single enantiomer (R) or (S) or enriched with one enantiomer.

3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from, in, L, R1, R2, and R3 are as defined in claim 2.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein, R1 is selected from 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-3-6-membered heterocyclic alkyl, wherein the 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-3-6-membered heterocyclic alkyl groups are each independently and optionally surrounded by 1, 2, or 3 R groups. a replace.

5. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein, R1 is selected from fluorine, chlorine, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, The CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, Each can be independently selected by 1, 2 or 3 Rs. a replace.

6. The compound according to claim 5 or a pharmaceutically acceptable salt thereof, wherein, R1 is selected from CH3, CF3, CH2CH3, CH2CHF2, CH2CF3, 7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein, R1 is selected from 8. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein, R2 is selected from H and CH3, wherein the CH3 is optionally divided by 1, 2 or 3 Rs. b replace.

9. The compound of claim 7 or a pharmaceutically acceptable salt thereof, wherein, R2 is selected from H, CH3, CF3, and CHF2.

10. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein, R3 is selected from H and CH3.

11. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein, R4 is selected from H and CH3.

12. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein, L is selected from single bonds and O.

13. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein, Structural unit Selected from 14. The compound of claim 13 or a pharmaceutically acceptable salt thereof, wherein, Structural unit Selected from 15. The following compounds or their pharmaceutically acceptable salts, 16. The compound of claim 15 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from, 17. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 16 in the preparation of a medicament related to complement factor B.

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