Reactive aldehyde species inhibitor designed on basis of ai, and use thereof

Abstract

Disclosed are a reactive aldehyde species (RASP) inhibitor designed on the basis of AI, and a use thereof. The RASP inhibitor comprises a compound represented by general formula (I) or a derivative thereof, wherein R1 and R2 are each independently selected from any one of hydrogen, a C2-6 unsaturated hydrocarbon group, a C1-6 alkoxy group, a C2-6 haloalkyl group, halogen, an amino group, a C3-20 cycloalkyl group, or a heterocyclic group containing 4-20 ring atoms; X is carbon or nitrogen; the derivative comprises at least one of a fluorescent label, a spin label, a heavy metal label, an isotope label, and a pharmaceutically acceptable salt. The provided RASP inhibitor has excellent capability and speed in sequestering aldehydes, and when applied to a lesion site, can quickly and effectively sequester RASP, to reduce the RASP level at the lesion site, thereby diminishing or preventing ocular inflammation. The RASP inhibitor shows potential preventive or therapeutic effects in various ocular inflammatory diseases such as dry eye disease.

Description

Active aldehyde substance inhibitor based on AI design and application thereof TECHNICAL FIELD

[0001] The present application belongs to the field of drug synthesis, and relates to an aldehyde trapping agent, in particular to an active aldehyde substance inhibitor based on AI design and application thereof. BACKGROUND

[0002] Dry eye disease (DED) refers to a series of diseases caused by various factors, such as abnormality of tear quality and quantity or dynamics, which leads to a decrease in tear film stability and is accompanied by ocular discomfort or ocular surface tissue lesions. The pathogenesis of dry eye is complex, and environmental, lifestyle, immunological disease, eye surgery, drug and age-related factors can all cause dry eye. Epidemiological surveys show that the incidence of dry eye in the world reaches 8%-34%, and the incidence of dry eye in Asia ranks the forefront in the world, among which the incidence in China reaches 21%-30%.

[0003] Reactive aldehyde species (RASP, Reactive aldehyde species), such as malondialdehyde (MDA) and 4-hydroxy-2-nonenal (HNE), can be generated in vivo through different physiological processes. They covalently bind to the amino and sulfhydryl groups of receptors and kinases, thereby enhancing the cascade of upstream pro-inflammatory signals involving NF-kB, inflammasome, scavenger receptor A, and other mediators (Higdon, A. et al., Biochem. J., 2012, 442:453-464; kalariya, N.M. et al., Exp. Eye Res., 2008, 86:70-80; Kauppinen, A. et al., Immunol. Lett., 2012, 147:29-33; Sapkota, M. et al., Respir. Res., 2017, 18:36). Elevated levels of RASP are found in various inflammatory eye diseases, including Behcet’s disease, Sjogren’s syndrome, non-infectious uveitis, allergic conjunctivitis, and dry eye (Sandikci, R. et al., Acta. Derm. Venereol., 2003, 83:342-346; Cejkova, J. et al., Mol. Vis., 2011, 17:443-447; Turk, A. et al., Ocul. Immunol. Inflamm., 2014, 22:127-132; Bacsi, A. et al., Allergy Clin. Immunol., 2005, 116:836-843; Choi, W. et al., Curr. Eye Res., 2016, 41:1143-1149). Studies found that MDA content in the tears of dry eye patients was elevated, and the degree of MDA elevation was positively correlated with the severity of dry eye (Augustin, A.J., et al., 1995, 233:694-698). Another study also showed that the levels of MDA and HNE were elevated in the tears and conjunctival tissue biopsies of dry eye patients compared to controls, and were correlated with the severity of symptoms (Choi, W. et al., Curr. Eye Res., 2016, 41:1143-1149). In addition to pro-inflammatory signals, RASP also binds to phosphatidylethanolamine. Phosphatidylethanolamine is a key component of the tear lipid group, which is essential for the moisture retention of ocular surface tissues (Butovich, I.A., et al., Prog. Lipid Res., 2011, 50:278-301). Therefore, RASP can be a potential important therapeutic target for treating dry eye.

[0004] RASP is closely related to the pathogenesis of various diseases, and has been confirmed as a target of major diseases such as dry eye, allergic conjunctivitis, uveitis, and Sjogren's syndrome. Therefore, RASP inhibitors have broad application prospects as drugs. However, so far there is no suitable RASP inhibitor on the market, and the existing RASP inhibitors in the clinical research stage are also few. Therefore, it is of great theoretical significance and application value to find RASP inhibitors with higher RASP inhibitory activity and lower toxicity. SUMMARY

[0005] The purpose of the present application is to provide a drug capable of inhibiting the production of active aldehyde substances in vivo, especially in the eye, to effectively treat or prevent dry eye.

[0006] To achieve the above-mentioned purpose, the present application provides an active aldehyde substance inhibitor comprising a compound having a general formula I or a derivative thereof,

[0007] wherein,

[0008] R1, R2 are independently selected from any one of hydrogen, C 2-6 unsaturated hydrocarbon group, C 1-6 alkoxy, C 2-6 halogenated alkyl, halogen, amino, C 3-20 cycloalkyl or heterocyclic group having 4-20 ring atoms;

[0009] X is carbon or nitrogen;

[0010] The derivative comprises at least one of a fluorescent marker, a spin marker, a heavy metal marker, an isotope marker and a pharmaceutically acceptable salt.

[0011] Optionally, R1 and R2 can form a C6-C 12 aromatic or non-aromatic ring together with the carbon atom to which they are connected.

[0012] Optionally, R1 and R2 can form a benzene ring or a fatty heterocyclic ring having 7-12 ring atoms together with the carbon atom to which they are connected.

[0013] Optionally, R1 and R2 are independently selected from any one of hydrogen, C 2-5 unsaturated hydrocarbon group, C 2-5 alkoxy and amino.

[0014] Optionally, R1 and R2 are independently selected from any one of hydrogen, C 6-12 cycloalkyl or heterocyclic group having 6-12 ring atoms.

[0015] Optionally, when R1 is halogen, R2 is C 2-6unsaturated hydrocarbon group.

[0016] Optionally, the compound of general formula I is selected from any one of:

[0017]

[0018] The present application also provides a pharmaceutical composition comprising the compound of general formula I or its derivative as an active ingredient.

[0019] Optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable pharmaceutical adjuvant.

[0020] Optionally, the pharmaceutical composition is a detection reagent or a kit comprising at least one of a fluorescent label, a spin label, a heavy metal label, an isotope label and a pharmaceutically acceptable salt of the compound of general formula I for identifying a RASP coordination base.

[0021] The present application also provides an application of the active aldehyde substance inhibitor as described above for preparing a medicine as an aldehyde capturing agent.

[0022] Optionally, the medicine is used for treating or preventing various diseases caused by active aldehyde substances.

[0023] Optionally, the disease refers to an ocular inflammatory disease.

[0024] Optionally, the ocular inflammatory disease comprises at least one of dry eye, allergic conjunctivitis, Behcet's disease, Sjogren's syndrome and uveitis.

[0025] Compared with the prior art, the technical scheme of the present application has at least the following beneficial effects:

[0026] The active aldehyde substance (RASP) inhibitor provided by the present application has good aldehyde complexing ability and speed, can quickly and effectively complex with active aldehyde substances when applied to a lesion site, reduces the content of RASP at the lesion site, and further reduces or avoids ocular inflammation, and has a potential preventive or therapeutic effect on various ocular inflammatory diseases such as dry eye. BRIEF DESCRIPTION OF DRAWINGS

[0027] FIG. 1 is a schematic diagram of the results of the efficacy test of the allergic conjunctivitis animal model of Example 28. DETAILED DESCRIPTION

[0028] As described in the foregoing background, the content of RASP in ocular or systemic inflammatory diseases increases, which can cause eye inflammation, reduced tear secretion and changed lipid composition in tears. For example, many dry eye patients have a high content of RASP. By inhibiting RASP, ocular inflammation can be reduced. Therefore, it is necessary to design a compound capable of inhibiting RASP.​

[0029] In the latest AI technology development, artificial intelligence can be used for the discovery, screening and optimization of small molecule drugs. By integrating AI computing tools, databases (such as CAS compound library) and receptor-ligand complex interaction information into an efficient molecular / atomic training set, AI digital workflow, and combining with BT technology (bioinformatics, in vitro target cell biological activity detection, etc.), new therapeutic drugs can be invented.

[0030] It is well known that in the process of drug development, traditional small molecule compounds (drugs) from lead compounds to lead compounds to candidate compounds require a lot of time (usually 5-6 years or even longer), and AI (AIDD) driven drug development can cross-compare data, molecular docking, molecular dynamics simulation, etc. in a shorter period of time (usually only 3-5 months), thereby accelerating the screening or de novo design of new compounds, and its core value lies in groundbreaking innovation and efficiency improvement.

[0031] Based on relatively clear mechanisms (such as Schrodinger equation, Gibbs free energy change, etc.), AI can search multiple more extensive and comprehensive different compound and proteomics database spaces, quickly complete search and cross-comparison, and save a lot of investment for wet experiments. Wet experiment data can be fed back to AI for iterative optimization of compound structure.

[0032] The present application is based on the AI (dry experiment) + BT (wet experiment) interactive fusion R&D mode of the self-built computing structural biology technology platform, combined with various drug chemistry, quantum chemistry frontier algorithms and deep learning, natural language processing (Natural Language Processing), various neural network framework pre-training models, to generate novel molecular structures with expected functional properties, and then perform molecular modeling, molecular docking and molecular dynamics simulation according to the target-drug related binding interface, etc. to obtain the optimal receptor-ligand binding free energy function, binding constant and dissociation constant, etc. to optimize and screen the in vitro cell activity optimal, comprehensive evaluation of the most suitable candidate compounds. At the same time, through disease / drug correlation network deep analysis, AI server and workstation, drug deep learning and multi-threaded collaborative simulation can be carried out synchronously and independently, or AI computing clusters can be formed to process library and candidate compound big data, deeply mine library, structure and pharmacophore relationship (QSAR) and new drug targets, save the cost of innovative drug development, and shorten the development time; thereby forming a new R&D path of "dry-wet combination, seamless connection" for new drug R&D, which is expected to become a scientific paradigm for future innovative drug R&D, and bring a new breakthrough in thinking and innovation in experiments to the past drug development process which is extremely dependent on time-consuming and laborious wet experiments.

[0033] The active aldehyde substance inhibitor according to the present application is screened and verified through multiple rounds of iterative wet-closed loop experiments, and finally obtained,

[0034] wherein,

[0035] R1, R2 are independently selected from hydrogen, C 2-6 unsaturated hydrocarbon group, C 1-6 alkoxy, C 2-6 haloalkyl, halogen, amino, C 3-20 cycloalkyl or heterocyclyl of 4-20 ring atoms;

[0036] X is carbon or nitrogen;

[0037] The derivative comprises at least one of a fluorescent label, a spin label, a heavy metal label, an isotope label and a pharmaceutically acceptable salt.

[0038] The "C 2-6 Unsaturated hydrocarbon group" refers to a straight-chain and branched-chain hydrocarbon group of 2 to 6 carbon atoms having at least one carbon-carbon double bond or carbon-carbon triple bond. Hydrocarbon group refers to a straight-chain or branched-chain group comprising several carbon atoms. The unsaturated hydrocarbon group can be substituted or non-substituted, and when substituted, the substituents thereof are preferably one or more substituents independently selected from one or more of deuterium atom, alkoxy, halogen, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0039] The "C 1-6 Alkoxy" refers to a group formed by the attachment of an alkyl group to an oxygen atom, wherein the oxygen atom has a free bonding capacity, such as methoxy, ethoxy, propoxy, butoxy, pentoxy, isopropoxy, tert-butoxy, cyclopropyloxy, cyclohexyloxy, etc. Alkoxy can be substituted or non-substituted, and when substituted, the substituents thereof are preferably one or more substituents independently selected from one or more of deuterium atom, alkoxy, halogen, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0040] The "C 1-6 Haloalkyl" includes monohaloalkyl and polyhaloalkyl (wherein all halogen atoms can be the same or different). Examples of haloalkyl include trifluoromethyl, 1,1-dichloroethyl, 1,2-dichloroethyl, 1,3-dibromo-3,3-difluoropropyl, etc.

[0041] "Heteroaryl" as used herein refers to a 5- to 20-membered, aromatic, monocyclic or multicyclic ring system containing one or more heteroatoms, each independently selected from the group consisting of O, N, and S, wherein the ring system can contain one or more double bonds, and wherein the ring system can be fused or bridged. Preferably, the heteroaryl group is 5- to 10-membered, more preferably 5- to 6-membered. Non-limiting examples of heteroaryl groups include:

[0042] "Amino" as used herein refers to -NH2. The amino group can be substituted or unsubstituted, and when substituted, the substituent is preferably one or more groups independently selected from one or more substituents selected from the group consisting of alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl.

[0043] "Cycloalkyl" as used herein refers to a saturated or partially unsaturated monocyclic or polycyclic ring system containing 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, preferably 3 to 10 carbon atoms (e.g., 3, 4, 5, 6, 7, 8, 9, or 10), more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyl groups include spirocyclic, fused, and bridged cycloalkyl groups.

[0044] "Sprio" as used herein refers to a 5- to 20-membered, polycyclic group sharing one carbon atom (referred to as a spiro atom) between single rings, which can contain one or more double bonds. Preferably, the spiro group is 6- to 14-membered, more preferably 7- to 10-membered (e.g., 7-, 8-, 9-, or 10-membered). The spirocycloalkyl group can be classified as a mono-, bi-, or polycycloalkyl group based on the number of spiro atoms shared between the rings, preferably a mono- or bi-cycloalkyl group. More preferably, the spirocycloalkyl group is a 3 / 5-, 3 / 6-, 4 / 4-, 4 / 5-, 4 / 6-, 5 / 5-, or 5 / 6- mono-cycloalkyl group. Non-limiting examples of spirocycloalkyl groups include:

[0045] at least one of the following:

[0046] "Fused" as used herein refers to a 5- to 20-membered, polycyclic group sharing an adjacent pair of carbon atoms between each ring in the structure, wherein one or more rings can contain one or more double bonds. Preferably, the fused group is 6- to 14-membered, more preferably 7- to 10-membered (e.g., 7-, 8-, 9-, or 10-membered). The fused cycloalkyl group can be classified as a bi-, tri-, tetra-, or polycycloalkyl group based on the number of rings, preferably a bi- or tri-cycloalkyl group, more preferably a 5 / 5- or 5 / 6- bi-cycloalkyl group. Non-limiting examples of fused cycloalkyl groups include:

[0047] at least one of the following:

[0048] "bridged ring" as used herein refers to a 5- to 20-membered, fully carbon polycyclic group in which any two rings share two non-adjacent carbon atoms, which can contain one or more double bonds. Preferably, it is 6- to 14-membered, more preferably 7- to 10-membered (e.g., 7-, 8-, 9-, or 10-membered). It can be a bicyclic, tricyclic, tetracyclic, or polycyclic bridged ring alkyl group, preferably a bicyclic, tricyclic, or tetracyclic, more preferably a bicyclic or tricyclic. Non-limiting examples of bridged ring alkyl groups include:

[0049] at least one of the following:

[0050] Polycyclic cycloalkyl groups can be substituted or unsubstituted, and when substituted, can be substituted at any available point of attachment with one or more substituents independently and optionally selected from the group consisting of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, and heteroaryl.

[0051] "Heterocyclyl" as used herein refers to a saturated or partially unsaturated monocyclic or polycyclic ring hydrocarbon substituent containing 4 to 20 ring atoms, one or more of which are heteroatoms selected from nitrogen, oxygen, and sulfur, which sulfur can optionally be oxidized (i.e., form a sulfoxide or sulfone), but not ring members of -O-O-, -O-S-, or -S-S-, with the remaining ring atoms being carbon. Preferably, it contains 7 to 12 ring atoms, one to four (e.g., one, two, three, and four) of which are heteroatoms; polycyclic heterocyclyl groups include spiro, fused, and bridged ring heterocyclyl groups.

[0052] "Spirocyclyl" as used herein refers to a 5- to 20-membered, polycyclic heterocyclic group in which the rings share a single atom (referred to as a spiro atom), one or more of which are heteroatoms selected from nitrogen, oxygen, and sulfur, which sulfur can optionally be oxidized (i.e., form a sulfoxide or sulfone), with the remaining ring atoms being carbon. It can contain one or more double bonds. Preferably, it is 6- to 14-membered. Spirocyclyl groups are classified as mono-, bi-, or polycyclic, preferably mono- and bi-cyclic, more preferably 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered spiroheterocyclyl groups. Non-limiting examples of spiroheterocyclyl groups include:

[0053] at least one of the following:

[0054] The term "fused heterocyclyl" as used herein refers to a 4- to 20-membered, polycyclic heterocyclic radical in which each ring shares an adjacent pair of atoms with another ring in the structure, one or more rings can contain one or more double bonds, one or more ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur, which can optionally be oxidized (i.e., form a sulfoxide or sulfone), and the remaining ring atoms are carbon. Preferably, the fused heterocyclyl is 6- to 14-membered, more preferably 7- to 10-membered (e.g., 7-, 8-, 9-, or 10-membered). Depending on the number of rings, the fused heterocyclyl can be bicyclic, tricyclic, tetracyclic, or polycyclic, preferably bicyclic or tricyclic, more preferably 3 / 4-, 3 / 5-, 3 / 6-, 4 / 4-, 4 / 5-, 4 / 6-, 5 / 4-, 5 / 5-, 5 / 6-, 6 / 3-, 6 / 4-, 6 / 5-, and 6 / 6 bicyclic fused heterocyclyl. Non-limiting examples of fused heterocyclyl groups include:

[0055] at least one of the following:

[0056] The term "bridged heterocyclyl" as used herein refers to a 5- to 14-membered, polycyclic heterocyclic radical in which any two rings share two non-adjacent atoms, which can contain one or more double bonds, one or more ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur, which can optionally be oxidized (i.e., form a sulfoxide or sulfone), and the remaining ring atoms are carbon. Preferably, the bridged heterocyclyl is 6- to 14-membered, more preferably 7- to 10-membered (e.g., 7-, 8-, 9-, or 10-membered). Depending on the number of rings, the bridged heterocyclyl can be bicyclic, tricyclic, tetracyclic, or polycyclic, preferably bicyclic, tricyclic, or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclyl groups include:

[0057] at least one of the following:

[0058] The polycyclic heterocyclyl group can be substituted or unsubstituted, and when substituted, the substituents can be substituted at any available point of attachment, and are preferably independently selected from one or more of the following: hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, and heteroaryl.

[0059] The term "aromatic ring" as used herein refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (fused polycyclic is sharing a pair of adjacent carbon atoms of the rings) ring having a conjugated pi-electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. The aromatic ring can be substituted or unsubstituted, and when substituted, it can be substituted at any available point of attachment with one or more substituents, preferably independently selected from one or more of saturated or unsaturated hydrocarbyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl.

[0060] The term "pharmaceutically acceptable salt" or "pharmaceutically acceptable salt" as used herein refers to salts of acids and bases that are pharmaceutically acceptable. Such pharmaceutically acceptable salts include salts of acids including hydrochloric, phosphoric, hydrobromic, sulfuric, sulfenic, formic, p-toluenesulfonic, methanesulfonic, nitric, benzoic, citric, tartaric, maleic, hydroiodic, chain carboxylic acids such as acetic acid, HOOC-(CH2) n -COOH (n = 0-4), and the like; and salts of bases having cations including sodium, potassium, calcium, ammonium, and the like.

[0061] In the present application, "substituted" means that an organic group defined herein (which contains one or more bonds to hydrogen atoms) is substituted with one or more bonds to non-hydrogen atoms or groups, i.e., substituents.

[0062] In the present application, "derivative" refers to a compound formed by substituting an atom or group in a compound of Formula (I) with another atom or group, including at least one of a fluorescent label, a spin label, a heavy metal label, an isotopic label, and a pharmaceutically acceptable salt.

[0063] In another aspect, the present application also relates to fluorescently, spin-labeled, heavy metal-labeled, or isotopically labeled derivatives of the compounds described herein, which are useful not only for imaging, but also for in vivo and in vitro detection, localization, and quantification of RASPs in tissue samples (including humans), and for identifying RASP binding groups (binding groups are groups that bind to the labeled compounds) by binding inhibition of the labeled compounds. Accordingly, the present application further provides RASP detection reagents or kits containing such labeled compounds.

[0064] The present application further provides isotopically-labeled compounds of the present application. An "isotopically-labeled compound" or "isotopically-labeled" of the present application means a compound described herein, wherein one or more atoms are replaced by an isotope atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature (i.e., naturally occurring). Suitable radioisotopes can include, but are not limited to, 2H (deuterium, also written as D), 3H (tritium, also written as T), 11C, 13C, 14C, 13N, 15N, 15O, 17O, 18O, 18F, 35S, 36Cl, 82Br, 75Br, 76Br, 77Br, 123I, 124I, 125I, and 131I. The radioisotope species contained in an isotopically-labeled compound will depend on the particular application of the isotopically-labeled compound. For example, for labeling and competition assays of IDO enzymes in vitro, compounds containing 3H, 14C, 82Br, 125I, 131I, 35S are generally most useful. For isotopic imaging applications, 11C, 18F, 125I, 123I, 124I, 131I, 75Br, 76Br, or 77Br are generally most useful.

[0065] Methods known in the art for labeling organic compounds with radioisotopes are equally applicable to the compounds of the present application.

[0066] When administered as a pharmaceutical, the compounds can be administered in the form of a pharmaceutical composition. Accordingly, in another aspect, the present application provides a pharmaceutical composition comprising a compound described herein, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.

[0067] The term "composition" as used herein is intended to encompass a product comprising the compound of the present application, or a pharmaceutically acceptable salt thereof, as the active ingredient, and any other component that is used in the formulation of the active ingredient.

[0068] Typically, the pharmaceutical composition contains at least one pharmaceutically acceptable carrier or excipient. The term "pharmaceutically acceptable" means the carrier or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. The carrier as used herein refers to a substance used to enhance the selectivity, effectiveness, and / or safety of a drug during delivery. The carrier is mainly used for controlling the release of the drug, and can also be used to improve the pharmacokinetic properties of the drug, particularly the bioavailability. The excipient refers to other substances in a pharmaceutical formulation in addition to the active ingredient, mainly for long-term stability, filling solid formulations (hence, also often used to specifically refer to "fillers") or enhancing the therapeutic efficacy of the product (e.g., promoting absorption, reducing viscosity, or increasing solubility, etc.).

[0069] The starting materials or reagents used in the embodiments of the present application are commercially available or prepared by synthetic methods generally known in the art.

[0070] The full names of the reagents used in the embodiments are represented by the abbreviations as follows:

[0071] Pd(dppf)Cl2: 1,1-bis(diphenylphosphino)ferrocene palladium dichloride

[0072] DMF: N,N-dimethylformamide

[0073] TMS: tetramethylsilane

[0074] TEA: triethylamine

[0075] NBS: N-bromosuccinimide

[0076] EA: ethyl acetate

[0077] PE: petroleum ether

[0078] LCMS detection: liquid chromatography and mass spectrometry combined detection

[0079] TLC detection: thin layer chromatography detection.

[0080] The technical solutions of the present application will be described clearly and completely in combination with the embodiments below. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor fall within the scope of protection of the present application.

[0081] Example 1: Preparation of compound 1

[0082] Synthetic route of compound 1:

[0083] Step 1: Preparation of intermediate 1a

[0084] To a solution of pyridine (1.5 mL) in methanol (25 mL) was added methyl 3-bromo-2-oxopropanoate (1.89 g, 10.45 mmol) dropwise at room temperature. The reaction was stirred at room temperature for 1 h and then at 60 °C for another 2 h. After the reaction was cooled to room temperature, 2-amino-5-bromobenzaldehyde (1.9 g, 9.50 mmol) and pyridine (5.0 mL) were added successively. The reaction mixture was heated to 85 °C and stirred for 4 h. Pyrrole (2.0 mL) was added and the temperature was maintained for another 4 h. The reaction was monitored by LCMS and was complete. The reaction was diluted with ethyl acetate (200 mL) and washed with saturated sodium chloride (50 mL x 3). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1 to 3:1) to give intermediate 1a (1.0 g, yield: 37.45%).

[0085] MS (ESI): m / z 283.0 [M+2+H] + .

[0086] Step 2: Preparation of intermediate 1b

[0087] To a solution of intermediate 1a (1.0 g, 3.56 mmol), cuprous iodide (67.8 mg, 0.356 mmol), and dichlorobis(triphenylphosphine)palladium (250 mg, 0.356 mmol) in 1,4-dioxane (30 mL) was added ethynyltrimethylsilane (1.05 g, 10.67 mmol) and triethylamine (2.5 mL, 17.8 mmol) successively under nitrogen protection. The reaction was stirred at 95 °C for 2 h.

[0088] The reaction was monitored by LCMS and was complete. The reaction was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1 to 4:1) to give intermediate 1b (1.0 g, yield: 94.34%).

[0089] MS (ESI): m / z 299.2 [M+H] + .

[0090] Step 3: Preparation of intermediate 1c

[0091] To a solution of intermediate 1b (1.0 g, 3.35 mmol) in methanol (20 mL) was added potassium carbonate (1.39 g, 10.05 mmol). The reaction was stirred at room temperature for 30 min.

[0092] The reaction was monitored by LCMS and was complete. The reaction was diluted with ethyl acetate (80 mL) and filtered through celite. The filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1 to 2:1) to give intermediate 1c (550 mg, yield: 72.5%).

[0093] MS (ESI): m / z 227.1 [M+H] + .

[0094] Step 4: Preparation of compound 1

[0095] Intermediate 1c (550 mg, 2.43 mmol) was dissolved in tetrahydrofuran (25 mL) and cooled to -70 °C, 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (4 mL, 12.15 mmol) was added dropwise, the reaction was maintained at temperature and stirred for 30 minutes.

[0096] LCMS indicated the reaction was complete, saturated ammonium chloride (20 mL) was added to the reaction and extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by prep (acetonitrile: 0.1% formic acid in water = 0% to 100%) to give compound 1 (100 mg, yield: 18.2%). MS (ESI): m / z 227.0 [M+H] + .

[0097] 1 H NMR (400 MHz, DMSO-d6) δ 7.76 (d, J = 2.0 Hz, 1H), 7.69 (d, J = 8.4 Hz, 1H), 7.30 (dd, Ji = 2.0 Hz, J2= 7.6 Hz, 1H), 7.20 (s, 1H), 6.02 (br s, 2H), 5.77 (br s, 1H), 1.60 (d, J = 2.8 Hz, 6H).

[0098] Example 2: Preparation of compound 2

[0099] Synthetic route of compound 2:

[0100] Step 1: Preparation of intermediate 2a

[0101] 5-Bromo-3-nitrobutyronitrile (25 g, 109.65 mmol) was dissolved in concentrated sulfuric acid (200 mL) and heated to 120 °C and stirred for 2 hours. The reaction was cooled to 0 °C and aqueous sodium nitrite (22.7 g, 329 mmol, dissolved in 50 mL water) was added dropwise. After the addition was complete, heating was continued to 80 °C and stirring was continued for 16 hours.

[0102] LCMS detection of reaction completion, the reaction solution was cooled to room temperature and poured into ice water (800 mL) to obtain a white suspension. After filtration, the filter cake was washed with water (600 mL). The filter cake was dried under vacuum to obtain intermediate 2a (25 g, yield: 92.3%). MS (ESI): m / z 247.0 [M+H] + .

[0103] Step 2: Preparation of intermediate 2b

[0104] To a solution of intermediate 2a (25 g, 101.2 mmol) in methanol (200 mL) was added concentrated sulfuric acid (15 mL). The reaction solution was stirred at 80 °C for 16 hours.

[0105] LCMS detection of reaction completion, the reaction solution was cooled to room temperature and poured into ice water (800 mL) to obtain a white suspension. After filtration, the filter cake was washed with water (600 mL). The filter cake was dried under vacuum to obtain intermediate 2a (25 g, yield: 92.3%). MS (ESI): m / z 247.0 [M+H]

[0106] MS (ESI): m / z 261.0 [M+H] + .

[0107] 1 H NMR (400 MHz, DMSO-d6) δ 9.15 (d, J = 2.0 Hz, 1H), 8.97 (d, J = 1.6 Hz, 1H), 3.93 (s, 3H).

[0108] Step 3: Preparation of intermediate 2c

[0109] Intermediate 2b (800 mg, 3.09 mmol), potassium vinyltrifluoroborate (828 mg, 6.18 mmol), 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (226 mg, 0.309 mmol), triethylamine (2.2 mL, 15.45 mmol) were dissolved in ethanol (20 mL) and replaced with nitrogen three times. The reaction solution was stirred at 90 °C for 2 hours.

[0110] LCMS detection of reaction completion, the reaction solution was cooled to room temperature and poured into ice water (800 mL) to obtain a white suspension. After filtration, the filter cake was washed with water (600 mL). The filter cake was dried under vacuum to obtain intermediate 2a (25 g, yield: 92.3%). MS (ESI): m / z 247.0 [M+H]

[0111] MS (ESI): m / z 261.0 [M+H] + .

[0112] Step 4: Preparation of intermediate 2d

[0113] Intermediate 2c (400 mg, 1.92 mmol) was dissolved in ethanol (20 mL), iron powder (538 mg, 9.6 mmol) and ammonium chloride (514 mg, 9.6 mmol) were added. The reaction was stirred at 50 °C for 1 h.

[0114] LCMS indicated the reaction was completed, the reaction was filtered, the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1 to 8:1) to give intermediate 2d (150 mg, yellow solid, yield: 43.9%).

[0115] MS (ESI): m / z 178.8 [M+H] + .

[0116] Step 5: Preparation of compound 2

[0117] Compound 2 was prepared from intermediate 2d (150 mg, 0.843 mmol) and 1.0 M methyl magnesium bromide in tetrahydrofuran (4.2 mL, 4.215 mmol) according to the method of Example 1, Step 4, yield: 21.3%.

[0118] MS (ESI): m / z 179.1 [M+H] + .

[0119] 1 H NMR (400 MHz, CD3OD) d 7.78 (d, J = 1.6 Hz, 1H), 7.17 (d, J = 1.6 Hz, 1H), 6.69 - 6.62 (m, 1H), 5.81 (dd, Ji = 1.2 Hz, J2 = 18.0 Hz, 1H), 5.81 (dd, Ji = 0.8 Hz, J2 = 8.4 Hz, 1H), 1.61 (s, 6H).

[0120] Example 3: Preparation of compound 3

[0121] Synthetic route of compound 3:

[0122] Step 1: Preparation of intermediate 3a

[0123] Intermediate 3a was prepared from methyl 3-amino-6-bromopyridine- carboxylate (1.0 g, 4.33 mmol) and potassium vinyltrifluoroborate (1.16 g, 8.66 mmol) according to the method of Example 2, Step 3, yield: 90.9%.

[0124] MS (ESI): m / z 179.2 [M+H] + .

[0125] Step 2: Preparation of compound 3

[0126] Compound 3 was prepared from intermediate 3a (200 mg, 1.12 mmol) and 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (3.7 mL, 11.2 mmol) following the procedure of Example 1, Step 4 in 45% yield.

[0127] MS (ESI): m / z 179.1 [M+H] + .

[0128] 1 H NMR (400 MHz, MeOD) d 7.07 (d, J = 8.0 Hz, 1H), 6.96 (d, J = 8.0 Hz, 1H), 6.65 (dd, J = 10.8 Hz, 17.6 Hz, 1H), 5.88 (d, J = 17.2 Hz, 1H), 1.60 (s, 6H).

[0129] Example 4: Preparation of compound 4

[0130] Synthetic route of compound 4:

[0131] Step 1: Preparation of intermediate 4a

[0132] Intermediate 4a was prepared from methyl 3-amino-5-bromopyridinate (1.2 g, 5.19 mmol) and ethynyltrimethylsilane (1.53 g, 15.6 mmol) following the procedure of Example 1, Step 2 in 82% yield.

[0133] MS (ESI): m / z 249.2 [M+H] + .

[0134] Step 2: Preparation of intermediate 4b

[0135] Intermediate 4b was prepared from intermediate 4a (1.5 g, 6.04 mmol) following the procedure of Example 1, Step 3 in 84.8% yield.

[0136] MS (ESI): m / z 177.1 [M+H] + .

[0137] Step 3: Preparation of compound 4

[0138] Compound 4 was prepared from intermediate 4b (150 mg, 0.852 mmol) and 1.0 M methyl magnesium bromide in tetrahydrofuran (4.3 mL, 4.26 mmol) following the procedure of Example 1, Step 4 in 53.7% yield.

[0139] MS (ESI): m / z 177.1 [M+H] + .

[0140] 1 H NMR (400 MHz, CD3OD) δ 7.80 (d, J = 2.0 Hz, 1H), 7.08 (d, J = 1.6 Hz, 1H), 3.55 (s, 1H), 1.57 (s, 6H).

[0141] Example 5: Preparation of compound 5

[0142] Synthetic route of compound 5:

[0143] Step 1: Preparation of intermediate 5a

[0144] Methyl 2-amino-5-fluorobenzoate (1.0 g, 5.92 mmol) was dissolved in acetonitrile (10 mL), and N-bromosuccinimide (1.26 g, 7.10 mmol) was added. Stirring at room temperature for 16 hours.

[0145] TLC detection showed that the reaction was completed, the reaction solution was diluted with water, extracted with ethyl acetate (20 mL x 3), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain intermediate 5a (1.0 g, yellow oil, crude product). The crude product was directly used in the next step reaction.

[0146] Step 2: Preparation of intermediate 5b

[0147] Intermediate 5b was prepared from intermediate 5a (500 mg, 2.01 mmol) and potassium vinyltrifluoroborate (402 mg, 3.02 mmol) according to the method of Example 2, step 3, in a yield of 66.0%.

[0148] MS (ESI): m / z 195.8 [M+H] + .

[0149] Step 3: Preparation of compound 5

[0150] Compound 5 was prepared from intermediate 5b (260 mg, 1.33 mmol) and 1.0 M methyl magnesium bromide in tetrahydrofuran (4.3 mL, 4.26 mmol) according to the method of Example 1, step 4, in a yield of 70.4%.

[0151] MS (ESI): m / z 178.1 [M+H-H2O] + .

[0152] 1H NMR (400 MHz, CD3OD) δ 6.95-6.83 (m, 3H), 5.59 (d, J = 17.2 Hz, 1H), 5.30 (dd, Ji = 0.8 Hz, J2= 10.8 Hz, 1H), 1.60 (s, 6H).

[0153] Example 6: Preparation of compound 6

[0154] Synthetic route of compound 6:

[0155] Step 1: Preparation of intermediate 6a

[0156] Intermediate 6a was prepared from methyl 2-amino-4-bromo-5-fluorobenzoate (500 mg, 2.0 mmol) and potassium vinyltrifluoroborate (536 mg, 4.0 mmol) following the procedure of Example 2, step 3 in 76.5% yield.

[0157] MS (ESI): m / z 195.8 [M+H] + .

[0158] Step 2: Preparation of compound 6

[0159] Compound 6 was prepared from intermediate 6a (300 mg, 1.54 mmol) and 1.0 M methylmagnesium bromide in tetrahydrofuran (7.7 mL, 7.7 mmol) following the procedure of Example 1, step 4 in 20% yield.

[0160] MS (ESI): m / z 178.1 [M+H-H20] + .

[0161] 1 H NMR (400 MHz, CD3OD) δ 6.86 (s, 1H), 6.83 (d, J = 6.8 Hz, 1H), 6.78-6.70 (m, 1H), 5.75 (dd, Ji = 1.2 Hz, J2= 17.6 Hz, 1H), 5.26 (dd, Ji = 1.2 Hz, J2= 11.2 Hz, 1H), 1.58 (s, 6H).

[0162] Example 7: Preparation of compound 7

[0163] Synthetic route of compound 7:

[0164] Step 1: Preparation of intermediate 7a

[0165] Intermediate 7a was prepared from methyl 2-amino-4-bromobenzoate (1.0 g, 4.34 mmol) and potassium vinyltrifluoroborate (750 mg, 5.64 mmol) following the procedure of Example 2, step 3 in 78% yield.

[0166] MS (ESI): m / z 178.2 [M+H] + .

[0167] 1 H NMR (400 MHz, CDCl3) δ 7.81 (d, J = 8.4 Hz, 1H), 6.74 (dd, J = 8.4 Hz, 1.6 Hz, 1H), 6.66 (d, J = 1.6 Hz, 1H), 6.65-6.58 (m, 1H), 5.79 (dd, J = 17.2 Hz, 0.4 Hz, 1H), 5.33 (dd, J = 10.8 Hz, 0.4 Hz, 1H), 3.87 (s, 3H).

[0168] Step 2: Preparation of compound 7

[0169] Compound 7 was prepared from intermediate 7a (350 mg, 1.97 mmol) and 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (3.95 mL, 11.85 mmol) following the procedure of Example 1, step 4 in 74.3% yield.

[0170] MS (ESI): m / z 160.1 [M+H-H20] + .

[0171] 1 H NMR (400 MHz, MeOD) δ 7.07 (d, J = 8.4 Hz, 1H), 6.78 (d, J = 1.6 Hz, 1H), 6.67 (dd, J = 1.6 Hz, 8.0 Hz, 1H), 6.58 (dd, J = 2.8 Hz, 17.6 Hz, 1H), 5.67 (dd, J = 1.2 Hz, 17.6 Hz, 1H), 5.11 (dd, J = 0.8 Hz, 10.8 Hz, 1H), 1.60 (s, 6H).

[0172] Example 8: Preparation of compound 8

[0173] Synthetic route of compound 8:

[0174] Step 1: Preparation of intermediate 8a

[0175] Intermediate 8a was prepared from methyl 2-amino-5-bromobenzoate (1.5 g, 6.52 mmol) and vinylpotassium trifluoroborate (1.3 g, 9.78 mmol) following the procedure of Example 2, step 3 in 78% yield.

[0176] MS (ESI): m / z 178.2 [M+H] + .

[0177] Step 2: Preparation of compound 8

[0178] Compound 8 was prepared from intermediate 8a (230 mg, 1.30 mmol) and 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (3.46 mL, 10.38 mmol) following the procedure of Example 1, step 4 in 65.2% yield.

[0179] MS (ESI): m / z 160.1 [M+H-H20] + .

[0180] 1 H NMR (400 MHz, MeOD) d 7.17 (d, J = 2.0 Hz, 1H), 7.10 (dd, J = 2.0 Hz, 8.0 Hz, 1H), 6.64 (d, J = 8.0 Hz, 1H), 6.57 (dd, J = 11.2 Hz, 17.6 Hz, 1H), 5.48 (dd, J = 0.8 Hz, 17.6 Hz, 1H), 4.93 (dd, J = 1.2 Hz, 10.8 Hz, 1H), 1.61 (s, 6H).

[0181] Example 9: Preparation of compound 9

[0182] Synthetic route for compound 9:

[0183] Step 1: Preparation of intermediate 9a

[0184] Intermediate 9a was prepared from methyl 2-amino-4-bromobenzoate (1.0 g, 4.35 mmol) and ethynyltrimethylsilane (1.28 g, 13.05 mmol) following the procedure of Example 1, step 2 in 93.1% yield.

[0185] MS (ESI): m / z 247.8 [M+H] + .

[0186] Step 2: Preparation of intermediate 9b

[0187] Intermediate 9b was prepared from intermediate 9a (1.0 g, 4.05 mmol) following the procedure of Example 1, step 3 in 99% yield.

[0188] MS (ESI): m / z 175.8 [M+H] + .

[0189] Step 3: Preparation of compound 9

[0190] Compound 9 was prepared from intermediate 9b (300 mg, 1.71 mmol) and 1.0 M methyl magnesium bromide in tetrahydrofuran (8.5 mL, 8.55 mmol) following the procedure of Example 1, step 4 in 71% yield.

[0191] MS (ESI): m / z 158.1 [M+H-H20] + .

[0192] 1 H NMR (400 MHz, CD3OD) δ 7.07 (d, J = 8.0 Hz, 1H), 6.78 (d, J = 1.6 Hz, 1H), 6.70 (dd, Ji = 1.6 Hz, J2= 8.0 Hz, 1H), 1.59 (s, 6H).

[0193] Example 10: Preparation of compound 10

[0194] Synthetic route for compound 10:

[0195] Step 1: Preparation of intermediate 10a

[0196] Intermediate 10a was prepared from 1-(2-amino-5-iodophenyl)ethan-1-one (700 mg, 2.69 mmol) and 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (904 mg, 5.38 mmol) following the procedure of Example 2, step 3 in 27.6% yield.

[0197] MS (ESI): m / z 176.2 [M+H] + .

[0198] Step 2: Preparation of compound 10

[0199] Compound 10 was prepared from intermediate 10a (130 mg, 0.743 mmol) and 1.0 M methyl magnesium bromide in tetrahydrofuran (3.7 mL, 3.715 mmol) following the procedure of Example 1, step 4 in 12.5% yield.

[0200] MS (ESI): m / z 174.1 [M+H-H20] + .

[0201] 1 H NMR (400 MHz, CD3OD) δ 7.08 (d, J = 1.6 Hz, 1H), 7.02 (dd, Ji = 2.0 Hz, J2= 8.4 Hz, 1H), 7.02 (dd, Ji = 1.2 Hz, J2= 15.6 Hz, 1H), 6.03 - 5.94 (m, 1H), 1.81 (dd, Ji = 1.6 Hz, J2= 6.8 Hz, 1H), 1.61 (s, 9H).

[0202] Example 11: Preparation of compound 11

[0203] Synthetic route of compound 11:

[0204] Step 1: Preparation of intermediate 11a

[0205] Methyl 2-amino-4-bromobenzoate (8.0 g, 34.72 mmol), 4,4,5,5-tetramethyl-2-(prop-l-en-2-yl)-l,3,2-dioxaborolane (8.75 g, 52.08 mmol), l,l-bis(diphenylphosphino)ferrocene palladium dichloride (2.54 g, 3.47 mmol), potassium carbonate (14.4 g, 104.16 mmol) were taken in 1,4-dioxane (80 mL) and water (8 mL) and purged with nitrogen gas for three times. The reaction was stirred at 90 °C for 2 h.

[0206] LCMS showed the reaction was completed and concentrated under reduced pressure. The crude was purified by silica gel column chromatography (PE:EA = 20:1 to 15:1) to give intermediate 11a (4.1 g, yellow oil, yield: 61.8%).

[0207] MS (ESI): m / z 192.2 [M+H] + .

[0208] Step 2: Preparation of compound 11

[0209] Compound 11 was prepared from intermediate 11a (4.1 g, 21.47 mmol) and 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (28.63 mL, 85.88 mmol) following the procedure of Example 1, step 4, yield: 39.4%.

[0210] MS (ESI): m / z 174.1 [M+H-H20] + .

[0211] 1 H NMR (400 MHz, CD3OD) δ 7.07 (d, J = 8.4 Hz, 1H), 6.84 (d, J = 2.0 Hz, 1H), 6.76 (dd, J1= 2.0 Hz, J2= 8.0 Hz, 1H), 5.29 (d, J = 0.8 Hz, 1H), 4.97 (t, J = 1.6 Hz, 1H), 2.07 (s, 3H), 1.60 (s, 6H).

[0212] Example 12: Preparation of compound 12

[0213] Synthetic route of compound 12:

[0214] Step 1: Preparation of intermediate 12a

[0215] Methyl 4-hydroxy-2-nitrobenzoate (600 mg, 3.043 mmol), 1-chloro-2-methoxyethane (863 mg, 9.13 mmol), potassium carbonate (1.26 g, 9.13 mmol) were dissolved in N,N-dimethylformamide (15 mL). The reaction solution was stirred at 80 °C for 5 hours.

[0216] LCMS detection showed that the reaction was completed, after the reaction solution was cooled to room temperature, diluted with ethyl acetate (150 mL), washed with saturated sodium chloride (20 mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, the crude product was purified by silica gel column chromatography (PE:EA = 10:1 to 4:1) to give intermediate 12a (520 mg, yellow oil, yield: 67.01%).

[0217] MS (ESI): m / z 273.1 [M+H2O] + .

[0218] Step 2: Preparation of intermediate 12b

[0219] Intermediate 12a (520 mg, 2.04 mmol), palladium on carbon (50 mg) were dissolved in methanol (15 mL). After the reaction solution was replaced with hydrogen, it was stirred at 60 °C for 16 hours.

[0220] LCMS detection showed that the reaction was completed, after the reaction solution was cooled to room temperature, the reaction solution was filtered through diatomite, and the filter residue was washed with ethyl acetate (30 mL). The filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA = 10:1 to 4:1) to give intermediate 12b (350 mg, yellow oil, yield: 76.25%).

[0221] MS (ESI): m / z 226.2 [M+H] +.

[0222] Step 3: Preparation of compound 12

[0223] Compound 12 was prepared from intermediate 12b (150 mg, 0.666 mmol) and 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (1.76 mL, 5.32 mmol) following the procedure of Example 1, Step 4 in a yield of 56.6%.

[0224] MS (ESI): m / z 208.1 [M+H-H20] + .

[0225] 1 H NMR (400 MHz, DMSO_d6) δ 6.86 (d, J = 8.4 Hz, 1H), 6.17 (d, J = 2.8 Hz, 1H), 6.04 (dd, J = 2.8 Hz, 8.8 Hz, 1H), 5.39 (brs, 2H), 5.07 (brs, 1H), 3.95 (t, J = 4.4 Hz, 2H), 3.60 (t, J = 4.8 Hz, 2H), 3.29 (s, 3H), 1.45 (s, 6H).

[0226] Example 13: Preparation of compound 13

[0227] Synthetic route of compound 13:

[0228] Step 1: Preparation of intermediate 13a

[0229] 6-chloro-2-methyl-3-nitropyridine (20.0 g, 116 mmol) was dissolved in concentrated sulfuric acid (100 mL) and cooled to 0 °C, then added with chromium trioxide (34.8 g, 348 mmol) in batches. Stirring at room temperature for 12 hours under nitrogen protection.

[0230] LCMS detected that the reaction was completed, the reaction solution was slowly poured into ice water (200 mL), and extracted with ethyl acetate (200 mL x 3). The organic phase was washed with saturated sodium chloride solution (200 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain intermediate 13a (19.0 g, green solid, yield: 81%).

[0231] MS (ESI): m / z 202.9 [M+H] + .

[0232] Step 2: Preparation of intermediate 13b

[0233] Intermediate 13a (19.0 g, 93.8 mmol) was dissolved in methanol (100 mL), and dichlorosulfoxide (10 mL) was added dropwise slowly. It was stirred at room temperature for 12 hours under nitrogen protection.

[0234] LCMS showed the reaction was completed, concentrated under reduced pressure, added ethyl acetate (100 mL), saturated aqueous sodium bicarbonate solution (50 mL), and extracted with ethyl acetate (50 mL x 3). The organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (PE:EA = 100:1 to 20:1) to give intermediate 13b (8.60 g, yellow oil, yield: 42%).

[0235] MS (ESI): m / z 217.0 [M+H] + .

[0236] Step 3: Preparation of intermediate 13c

[0237] Intermediate 13c was prepared from intermediate 13b (300 mg, 1.39 mmol) and 2-methoxyethan-1-ol (5 mL) according to the method of Example 12, Step 1, yield: 48%.

[0238] MS (ESI): m / z 300.80 [M+NH4] + .

[0239] Step 4: Preparation of intermediate 13d

[0240] Intermediate 13d was prepared from intermediate 13c (200 mg, 0.666 mmol) according to the method of Example 12, Step 2, yield: 89%.

[0241] MS (ESI): m / z 270.80 [M+H] + .

[0242] Step 5: Preparation of compound 13

[0243] Compound 13 was prepared from intermediate 13d (160 mg, 0.592 mmol) and 3.0 M methylmagnesium bromide in 2-methyltetrahydrofuran (2 mL, 6 mmol) according to the method of Example 1, Step 4, yield: 28%.

[0244] MS (ESI): m / z 227.2 [M+H] + .

[0245] 1H NMR (400 MHz, DMSO-d6) δ 6.99 (d, J = 8.4 Hz, 1H), 6.45 (d, J = 8.8 Hz, 1H), 5.31 (br s, 1H), 5.04 (br s, 2H), 4.21 (t, J = 4.8 Hz, 2H), 3.60 (t, J = 4.8 Hz, 2H), 3.27 (s, 3H), 1.46 (s, 6H).

[0246] Example 14: Preparation of compound 14

[0247] Synthetic route of compound 14:

[0248] Step 1: Preparation of intermediate 14a

[0249] Intermediate 14a was prepared from methyl 5-hydroxy-3-nitropyridine- carboxylate (500 mg, 2.52 mmol) and 1-chloro-2-methoxyethane (1.19 g, 12.62 mmol) following the method of Example 12, step 1 in 31% yield.

[0250] MS (ESI): m / z 257.1 [M+H] + .

[0251] Step 2: Preparation of intermediate 14b

[0252] Intermediate 14b was prepared from intermediate 14a (200 mg, 0.78 mmol) following the method of Example 12, step 2 in 82.9% yield.

[0253] MS (ESI): m / z 227.2 [M+H] + .

[0254] Step 3: Preparation of compound 14

[0255] Compound 14 was prepared from intermediate 14b (130 mg, 0.575 mmol) and 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (1.9 mL, 5.75 mmol) following the method of Example 1, step 4 in 57.69% yield.

[0256] MS (ESI): m / z 227.2 [M+H] + .

[0257] 1H NMR (400 MHz, MeOD) δ 7.54 (d, J = 2.4 Hz, 1H), 7.28 (d, J = 2.4 Hz, 1H), 4.25-4.23 (m, 2H), 3.77-3.75 (m, 2H), 3.41 (s, 3H), 1.66 (s, 6H).

[0258] Example 15: Preparation of compound 15

[0259] Synthetic route of compound 15:

[0260] Step 1: Preparation of intermediate 15a

[0261] Intermediate 15a was prepared from methyl 6-chloro-3-nitropyridine- carboxylate (300 mg, 1.39 mmol) and (E)-2-(3-methoxypropyl-l-en-l-yl)-4,4,5,5- tetramethyl-l,3,2-dioxaborolane (412 mg, 2.08 mmol) following the method of Example 11, step 1 in 77% yield.

[0262] MS (ESI): m / z 252.8 [M+H] + .

[0263] Step 2: Preparation of intermediate 15b

[0264] Intermediate 15b was prepared from intermediate 15a (270 mg, 1.07 mmol) following the method of Example 2, step 4 in 29% yield.

[0265] MS (ESI): m / z 222.8 [M+H] + .

[0266] Step 3: Preparation of compound 15

[0267] Compound 15 was prepared from intermediate 15b (70 mg, 0.315 mmol) and 3.0 M methylmagnesium bromide in 2-methyltetrahydrofuran (1 mL, 3 mmol) following the method of Example 1, step 4 in 28% yield.

[0268] MS (ESI): m / z 223.1 [M+H] + .

[0269] 1H NMR (400 MHz, DMSO-d6) δ 7.04 (d, J = 8.0 Hz, 1H), 6.89 (d, J = 8.0 Hz, 1H), 6.46 (d, J = 16 Hz, 1H), 6.39 - 6.32 (m, 1H), 5.62 (br s, 2H), 5.45 (br s, 1H), 4.01 (d, J = 5.6 Hz, 2H), 3.25 (s, 3H), 1.49 (s, 6H).

[0270] Example 16: Preparation of compound 16

[0271] Synthetic route of compound 16:

[0272] Step 1: Preparation of intermediate 16a

[0273] Intermediate 16a was prepared from 1-(2-amino-5-bromophenyl)ethanone (2.5 g, 11.7 mmol) and (E)-2-(3-methoxypropyl-l-en-l-yl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (2.54 g, 12.8 mmol) following the method of Example 11, Step 1 in 75% yield.

[0274] MS (ESI): m / z 206.2 [M+H] + .

[0275] Step 2: Preparation of compound 16

[0276] Compound 16 was prepared from intermediate 16a (1.80 g, 8.77 mmol) and 3.0 M methylmagnesium bromide in 2-methyltetrahydrofuran (23.4 mL, 70.2 mmol) following the method of Example 1, Step 4 in 43% yield.

[0277] MS (ESI): m / z 204.1 [M-H20+H] + .

[0278] 1 H NMR (400 MHz, CD3OD) δ 7.17 (s, 1H), 7.11 (d, J = 8.0 Hz, 1H), 6.65 (d, J = 8.4 Hz, 1H), 6.50 (d, J = 15.6 Hz, 1H), 6.08 - 6.00 (m, 1H), 4.04 (d, J = 6.4 Hz, 2H), 3.35 (s, 3H), 1.62 (s, 6H).

[0279] Example 17: Preparation of compound 17

[0280] Synthesis route of compound 17:

[0281] Step 1: Preparation of intermediate 17a

[0282] Methyl 5-fluoro-2-nitrobenzoate (5.0 g, 25.1 mmol), potassium carbonate (10.4 g, 75.3 mmol), 7-azaspiro[3.5]nonane hydrochloride (4.87 g, 30.1 mmol) were dissolved in N,N-dimethylformamide (50 mL). The reaction was stirred at 80 °C for 12 h.

[0283] LCMS showed the reaction was completed. Water (50 mL) was added and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (50 mL), the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1 to 3: 1) to give intermediate 17a (5.0 g, yellow solid, yield: 65%).

[0284] MS (ESI): m / z 305.2 [M+H] + .

[0285] Step 2: Preparation of intermediate 17b

[0286] Intermediate 17b was prepared from intermediate 17a (5.0 g, 16.4 mmol) following the procedure of Example 12, Step 2, yield: 67%.

[0287] MS (ESI): m / z 275.2 [M+H] + .

[0288] Step 3: Preparation of compound 17

[0289] Compound 17 was prepared from intermediate 17b (3.0 g, 10.9 mmol) and 3.0 M methylmagnesium bromide in 2-methyltetrahydrofuran (36.4 mL, 109 mmol) following the procedure of Example 1, Step 4, yield: 54%.

[0290] MS (ESI): m / z 275.1 [M+H] + .

[0291] 1H NMR (400 MHz, MeOD) δ 7.07 (d, J = 8.0 Hz, 1H), 6.77 (s, 1H), 6.69 (d, J = 8.0 Hz, 1H), 6.48 (d, J = 16.0 Hz, 1H), 6.24 - 6.17 (m, 1H), 4.05 (d, J = 6.0 Hz, 2H), 3.35 (s, 3H) 1.59 (s, 6H).

[0292] Example 18: Preparation of compound 18

[0293] Synthetic route for compound 18:

[0294] Step 1: Preparation of intermediate 18a

[0295] Intermediate 18a was prepared from methyl 2-amino-4-bromobenzoate (2.4 g, 10.4 mmol) and (E)-2-(3-methoxypropyl-l-en-l-yl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (2.47 g, 12.48 mmol) following the method of Example 11, Step 1 in 75.8% yield.

[0296] MS (ESI): m / z 222.1 [M+H] + .

[0297] Step 2: Preparation of compound 18

[0298] Compound 18 was prepared from intermediate 18a (1.75 g, 7.9 mmol) and 3.0 M methylmagnesium bromide in 2-methyltetrahydrofuran (21.1 mL, 63.2 mmol) following the method of Example 1, Step 4 in 82.6% yield.

[0299] MS (ESI): m / z 204.1 [M+H] + .

[0300] 1 H NMR (400 MHz, MeOD) δ 7.07 (d, J = 8.0 Hz, 1H), 6.77 (s, 1H), 6.69 (d, J = 8.0 Hz, 1H), 6.48 (d, J = 16.0 Hz, 1H), 6.24 - 6.17 (m, 1H), 4.05 (d, J = 6.0 Hz, 2H), 3.35 (s, 3H) 1.59 (s, 6H).

[0301] Example 19: Preparation of compound 19

[0302] Synthetic route for compound 19:

[0303] Step 1: Preparation of intermediate 19a

[0304] Intermediate 19a was prepared from 1-(2-amino-5-bromophenyl)ethanone (4.0 g, 18.7 mmol) and cyclohex-1-en-1-ylborane diol (3.53 g, 28.05 mmol) following the procedure of Example 11, Step 1 in 87% yield.

[0305] MS (ESI): m / z 216.3 [M+H] + .

[0306] Step 2: Preparation of intermediate 19b

[0307] To a solution of intermediate 19a (3.5 g, 16.28 mmol) in EtOAc (20 mL) was added Pd / C (350 mg). The reaction mixture was stirred at 20 °C under H2for 30 min.

[0308] LCMS indicated the reaction was completed, the reaction solution was filtered through celite, the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 20:1 to 10:1) to give intermediate 19b (2.5 g, yellow solid, yield: 70.8%).

[0309] MS (ESI): m / z 218.6 [M+H] + .

[0310] Step 3: Preparation of compound 19

[0311] Compound 19 was prepared from intermediate 19b (2.5 g, 11.5 mmol) and 3.0 M methylmagnesium bromide in 2-methyltetrahydrofuran (19.2 mL, 57.5 mmol) following the procedure of Example 1, Step 4 in 42.61% yield.

[0312] MS (ESI): m / z 234.1 [M+H] + .

[0313] 1 H NMR (400 MHz, MeOD) d 6.96 (s, 1H), 6.85 (d, J = 8.4 Hz, 1H), 6.63 (d, J = 8.0 Hz, 1H), 2.35 (s, 1H), 1.80 - 1.71 (m, 4H), 1.60 (s, 6H), 1.44 - 1.24 (m, 6H).

[0314] Example 20: Preparation of compound 20

[0315] Synthesis route of compound 20:

[0316] Step 1: Preparation of intermediate 20a

[0317] Methyl 5-fluoro-2-nitrobenzoate (700 mg, 3.515 mmol), potassium carbonate (1.457 g, 10.55 mmol), dimethylamine hydrochloride (458.61 mg, 5.624 mmol) were dissolved in N,N-dimethylformamide (10 mL). The reaction was stirred at 60 °C for 1 h.

[0318] LCMS detected that the reaction was completed, the reaction was diluted with ethyl acetate (120 mL), washed with saturated sodium chloride (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give intermediate 20a (600 mg, yellow oil, yield: 76.1%).

[0319] MS (ESI): m / z 247.1 [M+Na] + .

[0320] Step 2: Preparation of intermediate 20b

[0321] To a solution of intermediate 20a (600 mg, 2.676 mmol) in methanol (20 mL) was added Pd / C (284.78 mg, 0.2676 mmol). The reaction mixture was stirred at 50 °C under H2for 2 h.

[0322] LCMS detected that the reaction was completed, the reaction was filtered through celite, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1 to 4:1) to give intermediate 20b (450 mg, yellow solid, yield: 85.6%).

[0323] MS (ESI): m / z 195.2 [M+H] + .

[0324] Step 3: Preparation of compound 20

[0325] Compound 20 was prepared from intermediate 20b (400 mg, 2.06 mmol) and 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (5.5 mL, 16.5 mmol) according to the method of Example 1, step 4, yield: 40%.

[0326] MS (ESI): m / z 195.1 [M+H] + .

[0327] 1H NMR (400 MHz, MeOD) δ 6.73 (d, J = 38.4 Hz, 3H), 2.74 (s, 6H), 1.60 (s, 6H).

[0328] Example 21 : Preparation of compound 21

[0329] Synthetic route of compound 21:

[0330] Step 1: Preparation of intermediate 21a

[0331] Intermediate 21a was prepared from methyl 5-fluoro-2-nitrobenzoate (5.0 g, 25.1 mmol) and piperidine (4.28 g, 50.2 mmol) following the method of example 17, step 1 in 75% yield.

[0332] MS (ESI): m / z 264.80 [M+H] + .

[0333] Step 2: Preparation of intermediate 21b

[0334] Intermediate 21b was prepared from intermediate 21a (5.0 g, 18.9 mmol) following the method of example 12, step 2 in 90% yield.

[0335] MS (ESI): m / z 235.2 [M+H] + .

[0336] Step 3: Preparation of compound 21

[0337] Compound 21 was prepared from intermediate 21b (3.0 g, 12.8 mmol) and 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (42.7 mL, 128 mmol) following the method of example 1, step 4 in 47% yield.

[0338] MS (ESI): m / z 235.1 [M+H] + .

[0339] 1 H NMR (400 MHz, CD3OD) δ 6.90 (s, 1H), 6.78 (s, 1H), 6.68 (s, 1H), 2.93 (s, 4H), 1.80-1.72 (m, 4H), 1.61 (s, 6H), 1.55-1.48 (m, 2H).

[0340] Example 22: Preparation of compound 22

[0341] Synthetic route for compound 22:

[0342] Step 1: Preparation of intermediate 22a

[0343] Intermediate 22a was prepared from methyl 4-fluoro-2-nitrobenzoate (500 mg, 2.51 mmol) and dimethylamine hydrochloride (169.79 mg, 3.766 mmol) following the method of Example 17, step 1 in 84.38% yield.

[0344] MS (ESI): m / z 225.2 [M+H] + .

[0345] Step 2: Preparation of intermediate 22b

[0346] Intermediate 22b was prepared from intermediate 22a (500 mg, 2.23 mmol) following the method of Example 12, step 2 in 62.34% yield.

[0347] MS (ESI): m / z 195.0 [M+H] + .

[0348] Step 3: Preparation of compound 22

[0349] Compound 22 was prepared from intermediate 22b (150 mg, 0.7723 mmol) and 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (1.3 mL, 3.86 mmol) following the method of Example 1, step 4 in 23.92% yield.

[0350] MS (ESI): m / z 195.1 [M+H] + .

[0351] 1 H NMR (400 MHz, MeOD) d 6.97 (d, J = 7.6 Hz, 1H), 6.21 (d, J = 2.8 Hz, 1H), 6.14 (dd, J = 2.4, 8.8 Hz, 1H), 2.83 (s, 6H), 1.57 (s, 6H).

[0352] Example 23: Preparation of compound 23

[0353] Synthetic route for compound 23:

[0354] Step 1: Preparation of intermediate 23a

[0355] Intermediate 23a was prepared from methyl 4-fluoro-2-nitrobenzoate (500 mg, 2.51 mmol) and piperidine (427.58 mg, 5.02 mmol) following the procedure of Example 17, step 1 in 85.91% yield.

[0356] MS (ESI): m / z 265.2 [M+H] + .

[0357] Step 2: Preparation of intermediate 23b

[0358] Intermediate 23b was prepared from intermediate 23a (600 mg, 2.27 mmol) following the procedure of Example 12, step 2 in 94.13% yield.

[0359] MS (ESI): m / z 235.2 [M+H] + .

[0360] Step 3: Preparation of compound 23

[0361] Compound 23 was prepared from intermediate 23b (150 mg, 0.6402 mmol) and 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (1.1 mL, 3.2 mmol) following the procedure of Example 1, step 4 in 65% yield.

[0362] MS (ESI): m / z 235.1 [M+H] + .

[0363] 1 H NMR (400 MHz, MeOD) δ 6.99 (d, J = 8.8 Hz, 1H), 6.37 (d, J = 2.4 Hz, 1H), 6.30 (d, J = 2.8, 8.8 Hz, 1H), 3.06 (t, J = 5.4 Hz, 1H), 1.71 - 1.66 (m, 4H), 1.59 - 1.54 (m, 8H).

[0364] Example 24: Preparation of compound 24

[0365] Synthetic route for compound 24:

[0366] Step 1: Preparation of intermediate 24a

[0367] Intermediate 24a was prepared from ethyl 3,4-dihydroxybenzoate (4 g, 22 mmol) and piperidine 2-[2-(2-{[(4-methylphenyl)sulfonyl]oxy}ethoxy)ethoxy]-4- methylbenzenesulfonic acid ethyl ester (11.1 g, 24.2 mmol) following the procedure of Example 17, step 1 in 16.9% yield.

[0368] MS (ESI): m / z 297.2 [M+H] + .

[0369] Step 2: Preparation of Intermediate 24b

[0370] To a solution of intermediate 24a (1.1 g, 3.7 mmol) in acetic acid (15 mL) was added nitric acid (0.6 mL) and sulfuric acid (0.6 mL). The reaction was stirred at room temperature for 16 h.

[0371] LCMS indicated the reaction was complete, the reaction was diluted with ethyl acetate (200 mL), washed with saturated sodium chloride (50 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 3:1 to 1:1) to give intermediate 24b (950 mg, yellow solid, yield: 75.0%).

[0372] MS (ESI): m / z 342.2 [M+H] + .

[0373] Step 3: Preparation of Intermediate 24c

[0374] Intermediate 24c was prepared from intermediate 24b (950 mg, 2.78 mmol) following the procedure of Example 12, step 2 in 83.1% yield.

[0375] MS (ESI): m / z 312.2 [M+H] + .

[0376] Step 4: Preparation of Compound 24

[0377] Compound 24 was prepared from intermediate 24c (300 mg, 0.9636 mmol) and 3.0 M methyl magnesium bromide in 2-methyltetrahydrofuran (6.4 mL, 19.3 mmol) following the procedure of Example 1, step 4 in 8.73% yield.

[0378] MS (ESI): m / z 280.1 [M+H-H20] + .

[0379] 1H NMR (400 MHz, MeOD) δ 6.84 (s, 1H), 6.42 (s, 1H), 4.09-4.05 (m, 4H), 3.84-3.82 (m, 2H), 3.74-3.72 (m, 6H), 1.57 (s, 6H).

[0380] Example 25: Preparation of compound 25

[0381] Synthetic route for compound 25:

[0382] Step 1: Preparation of intermediate 25a

[0383] Intermediate 25a was prepared from 2-bromo-3-methylbut-2-ene (900 mg, 6.04 mmol) and methyl 2-amino-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)benzoate (2.175 g, 7.85 mmol) following the method of Example 11, Step 1 in 84.8% yield.

[0384] MS (ESI): m / z 220.2 [M+H] + .

[0385] Step 2: Preparation of compound 25

[0386] Compound 25 was prepared from intermediate 25a (400 mg, 1.82 mmol) and 1.0 M methylmagnesium bromide in tetrahydrofuran (18.2 mL, 18.2 mmol) following the method of Example 1, Step 4 in 70% yield.

[0387] MS (ESI): m / z 220.1 [M+H] + .

[0388] 1 H NMR (400 MHz, MeOD) δ 6.87 (s, 1H), 6.76 (d, J = 7.6 Hz, 1H), 6.66 (d, J = 8.0 Hz, 1H), 1.89 (s, 3H), 1.77 (s, 3H), 1.60 (s, 6H), 1.56 (s, 3H).

[0389] Example 26: Preparation of compound 26

[0390] Synthetic route for compound 26:

[0391] Step 1: Preparation of intermediate 26a

[0392] Intermediate 26a was prepared from 1-(2-amino-5-bromophenyl)ethanone (500 mg, 2.33 mmol) and 4,4,5,5-tetramethyl-2-(2-methylprop-1-en-1-yl)-1,3,2-dioxaborolane (552.86 mg, 3.037 mmol) following the procedure of Example 11, step 1 in 86.0% yield.

[0393] MS (ESI): m / z 190.2 [M+H] + .

[0394] Step 2: Preparation of compound 26

[0395] Compound 26 was prepared from intermediate 26a (380 mg, 1.85 mmol) and 1.0 M methyl magnesium bromide in tetrahydrofuran (9.2 mL, 9.2 mmol) following the procedure of Example 1, step 4 in 55.8% yield.

[0396] MS (ESI): m / z 188.1 [M+H-H20] + .

[0397] 1 H NMR (400 MHz, MeOD) d 6.99 (s, 1H), 6.88 (d, J = 8.0 Hz, 1H), 6.65 (d, J = 8.0 Hz, 1H), 1.84 (s, 3H), 1.81 (s, 3H), 1.60 (s, 6H).

[0398] In vitro activity evaluation

[0399] Example 27: In vitro aldehyde trapping capacity experiment

[0400] Main principle of the experiment:

[0401] The level of active aldehydes is found to be elevated in various inflammatory eye diseases, which, if not promptly eliminated, can exacerbate the symptoms of inflammation and worsen the eye disease. In this experiment, by simulating the in vivo environment, according to the complexing ability of the compound with nonenal, the relatively optimal compound is selected.

[0402] Experimental materials and reagents:

[0403] Sulfobutyl-β-cyclodextrin (Bide Pharmaceutica, BD243603-10g), linoleic acid (Aldrich, 69208F), triolein (Sigma-Aldrich, T7140-10G), nonenal (Bide Pharmaceutica, BD19638-1g).

[0404] Experimental steps:

[0405] S1, dissolve sulfobutyl-β-cyclodextrin (3 g) in phosphate buffer solution (12 ml) to prepare a solution. Use a pipette to transfer 2.88 ml of triolein and 2.88 ml of linoleic acid, 1:1, and mix well.

[0406] S2, at room temperature, add 48 μl of the above triolein / linoleic acid mixture to a 2 ml Eppendorf tube, add a dimethyl sulfoxide (5 μl) solution of the compound prepared in the examples of the present application (10 μmol, 1.0 equivalent), mix well, then add 100 μl of the above sulfobutyl-β-cyclodextrin solution, and finally add a dimethyl sulfoxide (10 μl) solution of nonenal (10 μmol, 1.0 equivalent), and stir the reaction at 1000 rpm.

[0407] S3, after 0 minutes (no nonenal solution added, 1510 μl of methanol added), 10 minutes, 20 minutes, 40 minutes and 60 minutes of stirring, respectively, add 1500 μl of methanol to the Eppendorf tube, high-speed vortex, then centrifuge quickly and transfer the solution to a sample bottle, and quantitatively monitor and analyze the residual amount of nonenal in the reaction mixture by high performance liquid chromatography.

[0408] Experimental results:

[0409] The liquid chromatography conditions are shown in Table 1 below:

[0410] Table 1: Liquid gradient conditions

[0411] Wherein, the sample injector temperature is 15°C; the column temperature is 40°C; and the nonenal UV-visible light detection wavelength is 220 nm.

[0412] The nonenal concentration at each time point (%) is shown in Table 2 below:

[0413] Table 2: Nonenal concentration at each time point (%) *: XRPR-5 is a similar control compound, Reproxalap.

[0414] When the nonenal concentration is lower (i.e., the nonenal residual amount is lower) at the same time point, it indicates that the compound has a stronger ability to complex aldehydes; when the nonenal concentration is consistent or comparable, the time point is smaller, indicating that the compound has a faster speed of complexing aldehydes. As can be seen from Table 2 above, except for Examples 5, 14 and 24, the compounds prepared in the examples of the present application all have very significant ability and speed of complexing aldehydes, and can be used to treat or prevent diseases with RASP-mediated pathological characteristics, including but not limited to dry eye syndrome, Behcet's disease, Sjogren's syndrome, non-infectious uveitis and allergic conjunctivitis, etc.

[0415] Animal efficacy evaluation

[0416] Example 28: Animal model of allergic conjunctivitis

[0417] Test principle and purpose: C48 / 80 is a mast cell degranulation agent that can directly bind to the receptors on the surface of mast cells, triggering the signal transduction pathway in mast cells. After mast cell degranulation, various inflammatory mediators are released. C48 / 80 and the inflammatory response it triggers can directly damage the epithelial barrier of the conjunctiva. Proteases and other substances released by inflammatory mediators and inflammatory cells can degrade the junction proteins between conjunctival epithelial cells, which can lead to damage to the integrity of the conjunctival epithelium, making the conjunctiva more susceptible to external pathogens, and also exacerbating the inflammatory response in the tissue. In addition, the inflammatory process stimulates nerve endings in the conjunctival tissue. On the one hand, inflammatory mediators such as histamine can activate sensory nerve endings, causing discomfort such as pain and itching. On the other hand, nerve endings stimulated will release neuropeptides such as substance P. Substance P has vasodilatory and pro-inflammatory effects, which can further exacerbate conjunctival hyperemia and inflammatory response. This test induces a rat conjunctivitis model by eye drops of C48 / 80, and investigates the improvement of the compound of the present application on the rat conjunctivitis model.

[0418] Sample preparation: The positive drug uses commercially available olopatadine hydrochloride eye drops (0.1%, Alcon, VEY89A); 1% compound 13 eye drops are prepared by hydrochloric acid, disodium hydrogen phosphate, sodium dihydrogen phosphate and water in a certain proportion.

[0419] Grouping: Female SD rats were used as research objects, and the SD rats were randomly grouped. The normal control group, the model control group, the olopatadine hydrochloride group and the compound 13 group were set up. The normal control group was not modeled, and the other groups were modeled. The specific grouping information is shown in Table 4.

[0420] Table 4: Drug and dose design table of each group

[0421] Modeling, drug administration and evaluation: The rats were grouped according to the random number method, and the rats were anesthetized. The normal control group was given 10 μL / eye of physiological saline to the conjunctival sac, and the other groups were given 10 μL / eye of C48 / 80 solution (source: MCE; specification and concentration: 50 mg / branch; batch number: 312920; 200 mg / mL, 0.9% physiological saline) to the conjunctival sac, and the eye was closed for 10 s to prevent the solution from overflowing. The modeling was completed.

[0422] 10 minutes after the completion of modeling, the first administration was performed: 15 μL / eye of the sample was taken and dropped into the conjunctival sac, and the eyelid was closed for 10 s to prevent the solution from overflowing. The normal control group and the model control group were given the same volume of normal saline. 20 minutes after the third administration and 24 hours after the completion of modeling, the eye examination was performed under the slit lamp, and the two eyes of each animal were scored according to the scoring rules in Table 5.

[0423] Table 5: Scoring rules for rat conjunctivitis

[0424] According to the scoring rules in Table 5, the score value is positively correlated with the severity of eye inflammation. The experimental results are shown in Figure 1, which proves that the compound 13 of the present application has a good therapeutic effect on the C48 / 80-induced allergic conjunctivitis of SD rats, wherein 1 represents the normal control group, 2 represents the model control group, 3 represents the olopatadine hydrochloride group, and 4 represents the compound 13 group.

[0425] Although the content of the present application has been described in detail through the above preferred embodiments, it should be recognized that the above description should not be considered as a limitation of the present application. After reading the above content, various modifications and alternatives of the present application will be apparent to those skilled in the art. Therefore, the protection scope of the present application should be defined by the appended claims.

Claims

1. An active aldehyde-based substance inhibitor designed based on AI, characterized in that, comprising a compound having the general formula (I) or a derivative thereof, wherein, R1, R2are independently selected from the group consisting of hydrogen, C 2-6 unsaturated hydrocarbon group, C 1-6 alkoxy, C 2-6 haloalkyl, halogen, amino, C 3-20 cycloalkyl or heterocyclyl having 4 to 20 ring atoms; X is carbon or nitrogen; The derivative comprises at least one of a fluorescent label, a spin label, a heavy metal label, an isotope label, and a pharmaceutically acceptable salt.

2. The AI-design-based active aldehyde substance inhibitor according to claim 1, wherein R1, R2can form together with the carbon atom to which they are attached a C6-C 12 aromatic or non-aromatic ring. 3.The AI-design-based active aldehyde substance inhibitor of claim 2, wherein, The R1 and R2 can form a benzene ring or a 7-12-membered aliphatic heterocyclic ring together with the carbon atoms to which they are connected.

4. The AI-design-based active aldehyde substance inhibitor according to claim 1, wherein said R1, R2are independently selected from the group consisting of hydrogen, C 2-5 unsaturated hydrocarbon group, C 2-5 alkoxy and amino.

5. The AI-design-based active aldehyde substance inhibitor according to claim 1, wherein R1, R2are independently selected from any one of hydrogen, C 6-12 Cycloalkyl or heterocyclyl having 6 to 12 ring atoms.

6. The AI-design-based active aldehyde substance inhibitor according to claim 1, wherein When R1is halogen, R2is C 2-6 unsaturated hydrocarbon group.

7. The AI-designed active aldehyde inhibitor as described in claim 1, characterized in that, The compounds of general formula (I) are selected from the group consisting of: Any one of the following:

8. A pharmaceutical composition, characterized by, The compound or derivative of the compound of general formula (I) according to any one of claims 1-7 is used as an active ingredient.

9. The pharmaceutical composition of claim 8, wherein, The pharmaceutical composition further comprises a pharmaceutically acceptable pharmaceutical adjuvant.

10. The pharmaceutical composition of claim 8, wherein The pharmaceutical composition is a detection reagent or a kit comprising at least one of a fluorescent label, a spin label, a heavy metal label, an isotope label, and a pharmaceutically acceptable salt of the compound of general formula (I) for identifying a RASP coordination base.

11. Use of an AI-designed active aldehyde-based inhibitor according to any one of claims 1-7, characterized in that, The pharmaceutical is used for preparing a medicine as an aldehyde capturing agent.

12. The use according to claim 11, wherein the compound is ###0005### or a pharmaceutically acceptable salt thereof. The medicine is used for treating or preventing various diseases caused by active aldehyde substances.

13. The use according to claim 12, wherein the compound is ###0002### The disease refers to an ocular inflammatory disease.

14. Use according to claim 13, wherein The ocular inflammatory disease comprises at least one of dry eye, allergic conjunctivitis, Behcet's disease, Sjogren's syndrome, and uveitis.

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