Ai design-based reactive aldehyde species inhibitor and use thereof

Abstract

Disclosed in the present invention is an AI design-based reactive aldehyde species inhibitor and the use thereof. The inhibitor contains a compound as shown in general formula (I) or a derivative thereof, wherein the definition of each group is as described in the description. The present invention is based on a self-built AI technical platform. By means of computationally predicting the basicity of aromatic amines, the transition-state activation energy △G of imine reactions and the enthalpy change △H of chemical reactions in combination with iterative optimization by means of repeated wet-lab experiments, the compound or the derivative thereof of the present invention is ultimately obtained. The compound or the derivative thereof can efficiently capture reactive aldehyde species (RASP), and can be used as an agent for capturing reactive aldehyde species in the treatment and / or prevention of various diseases caused by the reactive aldehyde species.

Description

Active aldehyde species 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 species inhibitor based on AI design and application thereof. BACKGROUND

[0002] Dry eye disease (DED) is a chronic ocular surface disease caused by abnormalities in tear quality, quantity, and dynamics, leading to tear film instability or ocular surface microenvironment imbalance, which can be accompanied by ocular surface inflammatory response, tissue damage, and nerve abnormalities, resulting in various uncomfortable symptoms and / or visual dysfunction in the eye. The causes of dry eye disease are complex, and environmental, lifestyle, immune diseases, eye surgery, drugs, and age-related factors can all cause dry eye disease. Epidemiological surveys show that the global incidence of dry eye is 8%-34%, and the incidence of dry eye in Asia ranks first in the world, among which the incidence in China has reached 21%-30%.

[0003] Active aldehyde species (RASP) such as malondialdehyde (MDA) and 4-hydroxy-2-nonenal (HNE) can be produced in vivo through different physiological processes. They can covalently bind to the amino and sulfhydryl groups of receptors and kinases, thereby enhancing the cascade reaction of upstream pro-inflammatory signaling pathways, which involve NF-kB, inflammasome, scavenger receptor A and other transmitters. 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 disease. Studies have found that the content of MDA in the tears of dry eye patients is increased, and the degree of MDA increase is positively correlated with the severity of dry eye disease. Another study also showed that compared with the control group, the levels of MDA and HNE in the tears and conjunctival tissue biopsy of dry eye patients were increased, and were correlated with the severity of symptoms. In addition to pro-inflammatory signaling pathways, RASP also binds to phosphatidylethanolamine. Phosphatidylethanolamine is a key component of tear lipid group, which is essential for water retention of ocular surface tissue. Therefore, RASP can be a potential important therapeutic target for the treatment of dry eye disease.

[0004] RASP is closely related to the pathogenesis of many diseases, and has been confirmed as a target of major diseases such as dry eye, allergic conjunctivitis, uveitis, and Sjogren's syndrome, so 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 clinical research are also few, so it is of great theoretical significance and application value to find RASP inhibitors with higher RASP inhibitory activity and lower toxicity. SUMMARY

[0005] In order to overcome the defects in the prior art, the present application prepares an active aldehyde substance inhibitor by AI technology, which has an unexpected inhibitory effect on active aldehyde substances.

[0006] [According to Rule 26, corrected on 27.10.2025] One object of the present application is to provide an active aldehyde substance inhibitor comprising a compound having the general formula (I) or a derivative thereof:

[0007] wherein R 1 is one or more substituents selected from hydrogen, hydroxyl, cyano, nitro, halogen, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcarbonyl and C 1-6 alkylamino;

[0008] any one;

[0009] R2, R3 are independently selected from H or CH3;

[0010] 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.

[0011] Optionally, R 1 is at least one selected from hydrogen, hydroxyl, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl and C 1-6 alkylamino.

[0012] Optionally, the compound having the general formula (I) is selected from at least one of:

[0013]

[0014] Another object of the present application is to provide a pharmaceutical composition comprising the above-mentioned compound having the general formula (I) or a derivative thereof as an active ingredient.​

[0015] Optionally, the pharmaceutical composition further comprises a pharmaceutical excipient.

[0016] 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 the RASP coordination base.

[0017] It is another object of the present application to provide the use of the compound of general formula (I) and derivatives thereof for the preparation of a medicament as an aldehyde trapping agent.

[0018] Optionally, the medicament can inhibit RASP for treating or preventing various diseases caused by active aldehyde substances.

[0019] Optionally, the disease refers to an ocular disease.

[0020] Optionally, the ocular disease comprises at least one of dry eye, allergic conjunctivitis, Behcet's disease, Sjogren's syndrome and uveitis. The uveitis refers to non-infectious uveitis.

[0021] Advantages of the present application

[0022] The present application prepares the compound of general formula (I) and unexpectedly finds that it has a surprisingly good RASP inhibiting effect, which can be used for preparing a medicament for treating or preventing diseases or conditions such as dry eye, Behcet's disease, Sjogren's syndrome, non-infectious uveitis and allergic conjunctivitis. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1 is a schematic diagram of the results of the pharmacodynamic experiment of compound 23 in the allergic conjunctivitis animal model of rats in Example 25 of the present application. DETAILED DESCRIPTION

[0024] The present application will be further illustrated by the following examples and figures, but the scope of protection of the present application is not limited to these examples. The percentages described in the present application are weight percentages unless otherwise specified. The numerical ranges described in the specification, such as measurement units, reaction conditions, physical state of the compound or percentage, are provided for clear and unambiguous written reference. Those skilled in the art can still obtain the expected results when using temperatures, concentrations, amounts, number of carbon atoms, etc. outside the range or different from the individual numerical values in the practice of the present patent.

[0025] 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), 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 are invented. It is well known that in the process of drug development, traditional small molecule compounds (drugs) from the lead compound to the lead compound to the candidate compound, a large amount of time (usually 5-6 years or even longer) is needed, and the AI (AIDrug Discovery & Design, AIDD) driven drug development can cross-compare data, molecular docking, molecular dynamics simulation, etc. in a short time (usually only 3-5 months), so as to accelerate the screening or de novo design of new compounds, and the core value lies in the original innovation and efficiency improvement.

[0026] Based on relatively clear mechanism (such as Schrodinger equation, Gibbs free energy change, etc.), AI can search multiple more extensive different compound and proteomics database space, quickly complete search and cross comparison, and save a lot of investment for wet experiment. Wet experiment data can be fed back to AI for iterative optimization of compound structure.

[0027] Based on NLP (Natural Language Processing) processing, disease / drug similarity network deep analysis, molecular generation model training, etc., the AI artificial intelligence server and workstation are independently built, which can synchronously and independently perform multiple drug deep learning and multi-thread cooperative simulation calculation, and can also be assembled into an AI computing cluster to process candidate compound big data, deeply mine new drug targets, save the cost of innovative drug development, and shorten the research and development time; so as to form a new research and development path of "dry-wet combination and seamless connection", which is expected to become a scientific paradigm for future innovative drug research and development, and bring a new breakthrough in thinking and innovation in experiment to the past drug research and development process which is extremely dependent on time-consuming and laborious wet experiment.

[0028] Based on the above-mentioned self-built AI technology platform, the basicity of aromatic amine, the activation energy ΔG of imine reaction transition state and the enthalpy change ΔH of chemical reaction are predicted, and then combined with wet experiment iterative optimization, a series of active aldehyde substance inhibitors in the present application are finally obtained.

[0029] The active aldehyde substance (RSAP) inhibitor provided by the present application comprises a compound represented by general formula (I) or a derivative thereof:

[0030] The basic principle of RSAP inhibitors to function is that the amino group in the structure reacts with the active aldehyde, thus reducing the concentration of the active aldehyde in the target tissue. The amino group reacts with the aldehyde to form an imine, and the reaction mechanism is as follows:

[0031] The aldehyde carbonyl group contains a carbon-oxygen double bond, and because the easily flowing π electron is strongly pulled towards the oxygen, the carbon of the carbonyl group is electron-deficient, and the oxygen of the carbonyl group is electron-rich. In the imine reaction, the lone pair of electrons of the electron-rich amine (base) attacks the electron-deficient carbon of the carbonyl group, forming an unstable imine alcohol, and one molecule of water is removed to form a carbon-nitrogen double bond (imine). The stronger the basicity of the amine, the easier the nucleophilic reaction. The basicity of the amine depends on the ability of the lone pair of electrons on the nitrogen to accommodate a positive charge. The amines in the present application are all aromatic amines. Certain substituents on the aromatic ring have an effect on the basicity of the amine. Electron-donating groups push electrons towards the nitrogen, increasing the basicity of the aromatic amine, and the lone pair of electrons can more effectively interact with the electron-deficient carbon; and electron-withdrawing groups help pull electrons away from the nitrogen, weakening the basicity of the aromatic amine, so that the lone pair of electrons is not easy to interact with the electron-deficient carbon. Common electron-donating groups include amino, hydroxyl, alkoxy, alkyl, phenyl, alkylamino, etc., and common electron-withdrawing groups include halogen, nitro, cyano, alkylcarbonyl, haloalkyl, etc. In the definition of R1, the substituents on the aromatic amines in embodiments 5, 6, and 7 are halogen, which is an electron-withdrawing group. These three embodiments have aldehyde capture ability. The cyano, nitro, C1-6 haloalkyl, and C1-6 alkylcarbonyl in the definition of R1 are all electron-withdrawing groups. It is theoretically reasonable to infer that compounds with such substituents have similar aldehyde capture ability to embodiments 5, 6, and 7; the hydroxyl, C1-6 alkyl, C1-6 alkoxy, and C1-6 alkylamino in the definition of R1 are all electron-donating groups. It is theoretically reasonable to infer that compounds with such substituents have better aldehyde capture ability than embodiments 5, 6, and 7.

[0032] Therefore, R 1 may be one or more substituents selected from hydrogen, hydroxyl, cyano, nitro, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcarbonyl, and C 1-6 alkylamino.

[0033] Y is selected from any one of -CH=CH-, -C≡ C-, -OCH2CH2O-, -OCH2CH=CH-, or -CH=CHCH2O-;

[0034] R2 and R3 are independently selected from H or CH3.

[0035] The definition of L in the compound provided by Chinese patent CN113227051 is a single bond, -O-, -S-, -NR2, and -(CR3R4)n-, all of which are electron-donating groups that increase the basicity of the aromatic amine and are conducive to imine reaction. The selected -CH=CH- and -C≡C- in the application are electron-withdrawing groups that reduce the basicity of the aromatic amine and are not conducive to imine reaction. In theory, the aldehyde capturing ability of the compound in the application should be weaker than that of the compound in patent CN113227051 when Y takes the above-mentioned electron-withdrawing groups, but the inventors of the application unexpectedly found that the aldehyde capturing ability of the compound in the application is stronger than that of the preferred compound in patent CN113227051, which has an unexpected technical effect.

[0036] The "halogen" described in the application refers to fluorine, chlorine, bromine, and iodine.

[0037] The "C 1-6 "Alkyl" refers to straight-chain and branched-chain or cyclic alkyl groups having 1 to about 6 carbon atoms. Examples of straight-chain alkyl groups include methyl, ethyl, n-propyl, n-butyl, n-pentyl, and n-hexyl. Examples of branched-chain alkyl groups include, but are not limited to, isopropyl, isobutyl, sec-butyl, t-butyl, neopentyl, isopentyl, isohexyl, and 2,2-dimethylpropyl. Examples of cyclic alkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0038] The "haloalkyl" described in the application includes monohaloalkyl and polyhaloalkyl (wherein all halogen atoms can be the same or different). Examples of haloalkyl groups include trifluoromethyl, 1,1-dichloroethyl, 1,2-dichloroethyl, 1,3-dibromo-3,3-difluoropropyl, and the like.

[0039] The "alkoxy" described in the application refers to a group formed by connecting the above-mentioned alkyl group to an oxygen atom, wherein the oxygen atom has a free bonding ability, such as methoxy, ethoxy, propoxy, butoxy, pentoxy, isopropoxy, tert-butoxy, cyclopropoxy, cyclohexyloxy, and the like.

[0040] The "pharmaceutically acceptable salt" or "pharmaceutically acceptable salt" described in the application refers to pharmaceutically acceptable acid and base addition salts and solvates. Such pharmaceutically acceptable salts include salts formed with acids, including hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, sulfurous acid, formic acid, p-toluenesulfonic acid, methanesulfonic acid, nitric acid, benzoic acid, citric acid, tartaric acid, maleic acid, hydroiodic acid, a chain carboxylic acid such as acetic acid, HOOC-(CH2) n -COOH (n=0-4), and the like; and salts formed with bases, the cations of which include sodium, potassium, calcium, ammonium ions, and the like.

[0041] ​In the present application, "substituted" means that an organic group as defined herein (containing one or more bonds to a hydrogen atom) is replaced by one or more bonds to a non-hydrogen atom or group, i.e., a substituent.

[0042] In the present application, "derivative" means a compound formed by substitution of an atom or group of atoms in a compound of Formula (I) with another atom or group of atoms, including at least one of a fluorescent label, a spin label, a heavy metal label, an isotopic label, and a pharmaceutically acceptable salt.

[0043] 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 sites by binding inhibition of the labeled compounds. Accordingly, the present application further provides RASP detection reagents or kits comprising such labeled compounds.

[0044] The present application further provides isotopically labeled compounds of the present application. An "isotopically labeled compound" or "isotopically labeled" compound of the present application refers to 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 radionuclides 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 kind of radioisotope contained in an isotopically labeled compound will depend on the particular use 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.

[0045] Methods known in the art for labeling organic compounds with radioisotopes are equally applicable to the compounds of the present application.

[0046] 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.

[0047] 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 product which results from combination of the active ingredient with one or more pharmaceutically acceptable carrier.

[0048] Generally, the pharmaceutical composition comprises 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 herein refers to a substance used to improve the selectivity, effectiveness and / or safety of a drug during delivery. The carrier is mainly used for controlling the release of a drug, and can also be used to improve the pharmacokinetic properties of a drug, particularly bioavailability. The excipient refers to a substance other than the active ingredient in a pharmaceutical preparation, which is mainly used for long-term stability, filling of solid preparations (therefore, it is often used to specifically refer to "filler") or enhancing the therapeutic effect of the product (e.g. promoting absorption, reducing viscosity or increasing solubility, etc.).

[0049] The starting materials or reagents used in the examples of the present application are commercially available or prepared by synthetic methods generally known in the art.

[0050] The full names of the reagents used in the examples are represented by the abbreviations as follows:

[0051] HATU: 2-(7-oxabenzotriazolyl)-N,N,N',N'-tetramethyluronium hexafluorophosphate

[0052] DIPEA: diisopropylethylamine

[0053] DABCO: 1,4-diazabicyclo[2.2.2]octane

[0054] HOBt: 1-hydroxybenzotriazole

[0055] EDCI.HCl: 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride

[0056] DMF: N,N-dimethylformamide

[0057] THF: tetrahydrofuran

[0058] EA: ethyl acetate

[0059] DCM: dichloromethane

[0060] PE: petroleum ether

[0061] LCMS detection: liquid chromatography-mass spectrometry combined detection

[0062] TLC detection: thin layer chromatography detection.

[0063] Example 1: Preparation of compound (E)-2,2'-(ethene-1,2-diylbis(3- aminopyridine-5,2-diyl))bis(propan-2-ol) (1)

[0064] Step 1: Preparation of compound 2-(3-amino-5-bromopyridin-2-yl)propan-2-ol (1a)

[0065] Methyl 3-amino-5-bromopyridinecarboxylate (2.5 g, 10.87 mmol) was dissolved in tetrahydrofuran (30 mL) and cooled to -78 °C, then 1.6 M methyl magnesium bromide tetrahydrofuran solution (27.2 mL, 43.48 mmol) was added slowly dropwise. The reaction was warmed to 0 °C and stirred for 3 hours under nitrogen protection. LCMS and TLC detection showed that the reaction was completed, then the reaction was slowly added saturated ammonium chloride aqueous solution (50 mL) at 0 °C, extracted with ethyl acetate (50 mL x 3), the organic phase was washed with saturated sodium chloride solution (50 mL) and dried over anhydrous sodium sulfate. Concentration under reduced pressure, the crude product was purified by silica gel column chromatography (PE:EA = 3:1, volume ratio) to give 2-(3-amino-5-bromopyridin-2-yl)propan-2-ol 1a (2.0 g, yield: 80%).

[0066] MS (ESI): m / z 231.1 [M+H] + .

[0067] Step 2: Preparation of compound 2-(3-amino-5-vinylpyridin-2-yl)propan-2-ol (1b)

[0068] 2-(3-amino-5-bromopyridin-2-yl)propan-2-ol 1a (600 mg, 2.6 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (400 mg, 2.6 mmol), 1,1-bis(diphenylphosphino)ferrocene palladium (II) dichloride (190 mg, 0.26 mmol) and cesium carbonate (2.1 g, 6.5 mmol) were dissolved in a mixed solution of 1,4-dioxane (20 mL) and water (4 mL). The reaction was stirred at 90 °C for 2 hours under nitrogen protection.

[0069] LCMS detection showed that the reaction was completed, then water (30 mL) was added to the reaction and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA = 5:1) to give 2-(3-amino-5-vinylpyridin-2-yl)propan-2-ol 1b (450 mg, yield: 95%).

[0070] MS (ESI): m / z 179.0 [M+H]+ .

[0071] Step 3: Preparation of compound (E)-2,2'-(ethene-1,2-diylbis(3-aminopyridine-5,2-diyl))bis(propan-2-ol) (1)

[0072] 2-(3-amino-5-vinylpyridin-2-yl)propanol 1b (300 mg, 1.68 mmol), 2-(3-amino-5-bromopyridin-2-yl)propan-2-ol (386 mg, 1.68 mmol), palladium(II) acetate (38 mg, 0.17 mmol) and potassium carbonate (580 mg, 4.2 mmol) were dissolved in 1,4-dioxane (10 mL). The reaction was stirred at 130 °C under nitrogen for 16 hours.

[0073] LCMS showed the reaction was completed, water (30 mL) was added to the reaction and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by preparation (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 10% to 60%) to give compound (E)-2,2'-(ethene-1,2-diylbis(3-aminopyridine-5,2-diyl))bis(propan-2-ol) 1 (37 mg, yield: 7%).

[0074] MS (ESI): m / z 329.1 [M+H] + .

[0075] 1 H NMR (400 MHz, DMSO-d6) δ 7.87 (d, J = 2.0 Hz, 2H), 7.17 (d, J = 2.0 Hz, 2H), 7.00 (s, 2H), 5.58 (s, 4H), 5.47 (s, 2H), 3.35 (s, 12H).

[0076] Example 2: Preparation of compound (E)-2-(3-amino-5-(2-(-5-amino-6-(2-hydroxypropan-2-yl)pyridin-2-yl)vinyl)pyridin-2-yl)propan-2-ol (2)

[0077] Step 1: Preparation of 2-(3-amino-6-bromopyridin-2-yl)propanol (2a)

[0078] Methyl 3-amino-6-bromopyridine-5-carboxylate (500 mg, 2.17 mmol) was dissolved in tetrahydrofuran (10 mL) and cooled to -78 °C, then 1.6 M methyl lithium tetrahydrofuran solution (5.4 mL, 8.68 mmol) was slowly added dropwise. The reaction was raised to 0 °C and stirred under nitrogen for 3 hours.

[0079] TLC detection reaction was completed, the reaction solution was slowly added saturated ammonium chloride aqueous solution (30 mL) at 0°C, extracted with ethyl acetate (30 mL x 3), washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate. Concentrated under reduced pressure, the crude product was purified by silica gel column chromatography (PE: EA = 15:1 to 2:1) to give 2-(3-amino-6-bromopyridin-2-yl)propanol 2a (350 mg, yield: 70%).

[0080] 1 H NMR (300 MHz, CDC13) δ 7.15 (d, J = 8.4 Hz, 1H), 6.83 (d, J = 8.1 Hz, 1H), 1.69 (s, 6H).

[0081] Step 2: Preparation of compound (E)-2-(3-amino-5-(2-(-5-amino-6-(2-hydroxypropan-2-yl)pyridin-2-yl)vinyl)pyridin-2-yl)propan-2-ol (2)

[0082] 2-(3-amino-6-bromopyridin-2-yl)propanol 2a (385 mg, 1.68 mmol), 2-(3-amino-5-vinylpyridin-2-yl)propanol 1b (300 mg, 1.68 mmol), palladium (II) acetate (38 mg, 0.17 mmol) and potassium carbonate (580 mg, 4.2 mmol) were dissolved in 1,4-dioxane (10 mL). The reaction solution was stirred at 130°C under nitrogen protection for 16 hours.

[0083] LCMS detection reaction was completed, water (30 mL) was added to the reaction solution and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by preparation (acetonitrile: 0.1% trifluoroacetic acid aqueous solution = 10% to 60%) to give (E)-2-(3-amino-5-(2-amino-6-(2-hydroxy-2-yl)pyridin-2-yl)vinyl)pyridin-2-yl)propanol 2 (49 mg, yield: 9%).

[0084] MS (ESI): m / z 329.1 [M+H] + .

[0085] 1H NMR (400 MHz, DMSO-d6) δ 8.01 (s, 1H), 7.77 (s, 1H), 7.34 (d, J = 7.6 Hz, 1H), 7.26 (s, 2H), 7.05 (d, J = 7.6 Hz, 1H), 5.49 (br s, 6H), 1.60 (s, 6H), 1.56 (s, 6H).

[0086] [According to Rule 26 Correction 27.10.2025] Example 3: Preparation of compound (E)-2,2'-(ethene-1,2-diylbis(3-aminopyridine-6,2-diyl))bis(propan-2-ol) (3)

[0087] Step 1: Preparation of 2-(3-amino-6-vinylpyridin-2-yl)propanol (3a)

[0088] 2-(3-amino-6-bromopyridin-2-yl)propanol (400 mg, 1.74 mmol), vinyl tetrafluoroborate (202 mg, 1.74 mmol), 1,1-bis(diphenylphosphino)ferrocene palladium(II) dichloride (124 mg, 0.17 mmol) and cesium carbonate (1.4 g, 4.35 mmol) were dissolved in a mixed solution of 1,4-dioxane (20 mL) and water (4 mL). The reaction was stirred at 90 °C under nitrogen protection for 2 hours.

[0089] LCMS detected that the reaction was completed, water (30 mL) was added to the reaction and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA = 5:1) to obtain 2-(3-amino-6-vinylpyridin-2-yl)propanol 3a (200 mg, yellow oil, yield: 65%).

[0090] MS (ESI): m / z 179.0 [M+H] + .

[0091] Step 2: Preparation of compound (E)-2,2'-(ethene-1,2-diylbis(3-aminopyridine-6,2-diyl))bis(propan-2-ol) (3)

[0092] 2-(3-amino-6-vinylpyridin-2-yl)propanol 3a (200 mg, 1.12 mmol), 2-(3-amino-6-bromopyridin-2-yl)propanol (258 mg, 1.12 mmol), palladium(II) acetate (25 mg, 0.11 mmol) and potassium carbonate (386 mg, 2.8 mmol) were dissolved in 1,4-dioxane (10 mL). The reaction was stirred at 130 °C under nitrogen protection for 16 hours.

[0093] LCMS detected completion of the reaction, water (10 mL) was added to the reaction solution and extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by preparation (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 10% to 60%) to obtain (E)-2,2'-(ethene-1,2-diylbis(3-aminopyridine-6,2-diyl))bis(propan-2-ol) 3 (35 mg, yield: 9.5%).

[0094] MS (ESI): m / z 329.1 [M+H] + .

[0095] 1 H NMR (400 MHz, DMSO-d6) δ 7.12 (d, J = 9.2 Hz, 4H), 6.91 (d, J = 8.0 Hz, 2H), 5.64 (s, 4H), 5.49 (s, 2H), 1.53 (s, 12H).

[0096] [According to Rule 26 Corrected 27.10.2025] Example 4: Preparation of compound (E)-2-(2-amino-4-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridin-3-yl)vinyl)phenyl)propan-2-ol (4)

[0097] Step 1: Preparation of methyl 2-amino-4-vinylbenzoate (4a)

[0098] Methyl 2-amino-4-bromobenzoate (1.0 g, 4.34 mmol) was dissolved in ethanol (20 mL), then potassium vinyltrifluoroborate (750 mg, 5.64 mmol), triethylamine (1.32 g, 13.0 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (317 mg, 0.434 mmol) were added. Stirring was performed under nitrogen protection at 90°C for 5 hours.

[0099] LCMS detected completion of the reaction, water (30 mL) was added to the reaction solution and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA = 10:1) to obtain methyl 2-amino-4-vinylbenzoate 4a (600 mg, yellow oil, yield: 78%).

[0100] MS (ESI): m / z 178.2 [M+H] + .

[0101] 1 H NMR (400 MHz, CDC13) 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).

[0102] Step 2: Preparation of methyl ((E)-3-amino-5-(3-amino-4- (methoxycarbonyl)styryl)pyridinecarboxylate (4b)

[0103] Methyl 2-amino-4-vinylbenzoate 4a (200 mg, 1.13 mmol), methyl 3-amino-5- bromopyridinecarboxylate (261 mg, 1.13 mmol), palladium (II) acetate (25 mg, 0.113 mmol) and potassium carbonate (312 mg, 2.23 mmol) were dissolved in 1,4- dioxane (10 mL). The reaction was stirred at 130 °C under nitrogen for 16 hours.

[0104] LCMS showed the reaction was completed, water (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, concentrated under reduced pressure, and the crude product was purified by reverse phase column (acetonitrile: water = 5% to 95%) to give methyl ((E)-3-amino-5-(3-amino-4- (methoxycarbonyl)styryl)pyridinecarboxylate 4b (130 mg, yellow oil, yield: 35%).

[0105] MS (ESI): m / z 328.2 [M+H] + .

[0106] Step 3: Preparation of (E)-2-(2-amino-4-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridin-3- yl)vinyl)phenyl)propan-2-ol (4)

[0107] Methyl ((E)-3-amino-5-(3-amino-4-(methoxycarbonyl)styryl)pyridinecarboxylate 4b (130 mg, 0.397 mmol) was dissolved in tetrahydrofuran (2 mL) and cooled to -78 °C, then 3M methyl magnesium bromide tetrahydrofuran solution (1.3 mL, 3.9 mmol) was added slowly dropwise. The reaction was raised to 0 °C and stirred for 2 hours under nitrogen.

[0108] LCMS monitoring of the reaction completion, the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by prep (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 40% to 75%) to give ((E)-2-(2-amino-4-(2-amino-6-(2-hydroxypropyl-2-yl)pyridin-3- ethenyl)-5-chlorophenyl)propan-2-ol 4 (50 mg, white solid, yield: 38%).

[0109] MS (ESI): m / z 328.1 [M+H] + .

[0110] 1 H NMR (400 MHz, CD3OD) δ 7.86 (d, J = 2.0 Hz, 1H), 7.24 (d, J = 1.6 Hz, 1H), 7.13 (d, J = 8.0 Hz, 1H), 7.04 (d, J = 16.4 Hz, 1H), 6.96 (d, J = 16.4 Hz, 1H), 6.90 (d, J = 2.0 Hz, 1H), 6.84 (dd, J = 8.0 Hz, 2.0 Hz, 1H), 1.62 (s, 6H), 1.61 (s, 6H).

[0111] [Amended according to Rule 26 - Corrected 27.10.2025] Example 5: Preparation of compound (E)-2-(2-amino-3-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridin-2-yl)vinyl)-5- fluorophenyl)propan-2-ol (5)

[0112] Step 1: Preparation of methyl 2-amino-5-fluoro-3-vinylbenzoate (5a)

[0113] Methyl 2-amino-4-bromo-5-fluorobenzoate (1.0 g, 4.03 mmol) was dissolved in ethanol (15 mL), then potassium vinyltrifluoroborate (804 mg, 6.05 mmol), triethylamine (2.24 mL, 16.12 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (295 mg, 0.403 mmol) were added. Stirring was carried out under nitrogen protection at 90°C for 2 hours.

[0114] LCMS monitoring of the reaction completion, the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by prep (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 40% to 75%) to give ((E)-2-(2-amino-4-(2-amino-6-(2-hydroxypropyl-2-yl)pyridin-3- ethenyl)-5-chlorophenyl)propan-2-ol 4 (50 mg, white solid, yield: 38%).

[0115] MS (ESI): m / z 196.2 [M+H] + .

[0116] Step 2: Preparation of (E)-methyl 2-amino-3-(2-(5-amino-6-(2-hydroxypropan-2- yl)pyridin-2-yl)vinyl)-5-fluorobenzoate (5b)

[0117] Methyl 2-amino-5-fluoro-3-vinylbenzoate 5a (338 mg, 1.73 mmol), 2-(3-amino-6- bromopyridin-2-yl)propan-2-ol (400 mg, 1.73 mmol), palladium(II)acetate (79 mg, 0.346 mmol) and potassium carbonate (717 mg, 5.2 mmol) were dissolved in 1,4-dioxane (15 mL). The reaction was stirred at 130 °C under nitrogen for 16 hours.

[0118] LCMS showed the reaction was completed, the reaction was concentrated under reduced pressure, the crude product was purified by silica gel column chromatography (PE:EA = 10:1-1:2) to give (E)-methyl 2-amino-3-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridin-2- yl)vinyl)-5-fluorobenzoate 5b (420 mg, yield: 70%).

[0119] MS (ESI): m / z 346.2 [M+H] + .

[0120] Step 3: Preparation of (E)-2-(2-amino-3-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridin-2- yl)vinyl)-5-fluorophenyl)propan-2-ol (5)

[0121] (E)-methyl 2-amino-3-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridin-2-yl)vinyl)-5- fluorobenzoate 5b (420 mg, 1.22 mmol) was dissolved in tetrahydrofuran (10 mL) and cooled to 0 °C, then 1.0 M methylmagnesium bromide in tetrahydrofuran (6.1 mL, 6.09 mmol) was added slowly dropwise. The reaction was stirred under nitrogen for 2 hours.

[0122] LCMS monitoring of the reaction completion, water (30 mL) was added to the reaction solution and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by preparative (acetonitrile: 0.1% trifluoroacetic acid aqueous solution = 30% to 60%) to give (E)-2-(2-amino-3-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridin-2-yl)vinyl)-5-fluorophenyl)propan-2-ol trifluoroacetate salt 5 (190 mg, yield: 34%).

[0123] MS (ESI): m / z 346.1 [M+H] + .

[0124] 1 H NMR (400 MHz, CD3OD) δ 7.88 (d, J = 8.8 Hz, 1H), 7.59-7.70 (m, 1H), 7.58 (d, J = 8.8 Hz, 1H), 7.34-7.41 (m, 2H), 7.08-7.11 (m, 1H), 1.72 (s, 6H), 1.65 (s, 6H).

[0125] [According to Rule 26 Corrected 27.10.2025] Example 6: Preparation of compound (E)-2-(2-amino-4-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridin-3-yl)vinyl)-5-fluorophenyl)propan-2-ol (6)

[0126] Step 1: Preparation of methyl 2-amino-5-fluoro-4-vinylbenzoate (6a)

[0127] Methyl 2-amino-4-bromo-5-fluorobenzoate (300 mg, 1.21 mmol) was dissolved in ethanol (10 mL), then potassium vinyltrifluoroborate (241 mg, 1.81 mmol), triethylamine (367 mg, 3.63 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (88 mg, 0.121 mmol) were added. Stirring was performed under nitrogen protection at 90°C for 5 hours.

[0128] LCMS monitoring of the reaction completion, water (30 mL) was added to the reaction solution and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA = 10:1) to give methyl 2-amino-5-fluoro-4-vinylbenzoate 6a (200 mg, yellow oil, yield: 85%).

[0129] MS (ESI): m / z 196.0 [M+H] + .

[0130] 1 H NMR (400 MHz, CDC13) δ 7.52 (d, J = 11.6 Hz, 1H), 6.80-6.73 (m, 2H), 5.87 (dd, J = 18.0 Hz, 0.8 Hz, 1H), 5.45 (d, J = 11.2 Hz, 1H), 3.87 (s, 3H).

[0131] Step 2: Preparation of methyl (E)-3-amino-5-(5-amino-2-fluoro-4- (methoxycarbonyl)styryl)pyridinecarboxylate (6b)

[0132] Methyl 2-amino-5-fluoro-4-vinylbenzoate 6a (200 mg, 1.02 mmol), methyl 3- amino-5-bromopyridinecarboxylate (237 mg, 1.02 mmol), palladium (II) acetate (23 mg, 0.102 mmol) and potassium carbonate (282 mg, 2.04 mmol) were dissolved in 1,4-dioxane (10 mL). The reaction was stirred at 130 °C under nitrogen atmosphere for 16 hours.

[0133] LCMS showed the reaction was completed, water (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, concentrated under reduced pressure, and the crude product was purified by reverse phase column (acetonitrile: water = 5% to 95%) to give methyl 3-amino-5-(5-amino-2-fluoro-4- (methoxycarbonyl)styryl)pyridinecarboxylate 6b (130 mg, yellow oil, yield: 37%).

[0134] MS (ESI): m / z 346.2 [M+H] + .

[0135] Step 3: Preparation of (E)-2-(2-amino-4-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridin-3- yl)vinyl)-5-fluorophenyl)propan-2-ol (6)

[0136] Methyl 3-amino-5-(5-amino-2-fluoro-4-(methoxycarbonyl)styryl)pyridinecarboxylate 6b (130 mg, 0.376 mmol) was dissolved in tetrahydrofuran (2 mL) and cooled to -78 °C, then 3M methyl magnesium bromide tetrahydrofuran solution (1.3 mL, 3.9 mmol) was added slowly dropwise. The reaction was raised to 0 °C and stirred under nitrogen atmosphere for 2 hours.

[0137] LCMS monitoring of the reaction completion, the reaction solution was added water (30 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by preparative (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 30% to 60%) to give (E)-2-(2-amino-4-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridin-3-yl)vinyl)-5-fluorophenyl)propan-2-ol 6 (40 mg, yield: 31%, NMR showed that the product contained about 25% of cis isomer).

[0138] MS (ESI): m / z 346.1 [M+H] + .

[0139] 1 H NMR (400 MHz, CD3OD) δ 7.86 (d, J = 2.0 Hz, 1H), 7.25 (d, J = 1.6 Hz, 1H), 7.18 (d, J = 16.4 Hz, 1H), 7.04 (d, J = 16.8 Hz, 1H), 6.98 (d, J = 6.8 Hz, 1H), 6.90 (d, J = 12.8 Hz, 1H), 1.61 (s, 6H), 1.60 (s, 6H).

[0140] [According to Rule 26 Corrected 27.10.2025] Example 7: Preparation of the synthesis of compound ((E)-2-(2-amino-4-(2-amino-6-(2-hydroxypropyl-2-yl)pyridin-3-ylvinyl)-5-chlorophenyl)propan-2-ol (7)

[0141] Step 1: Preparation of methyl 2-amino-4-bromo-5-chlorobenzoate (7a)

[0142] 2-amino-4-bromo-5-chlorobenzoic acid (1.5 g, 6 mmol) was dissolved in a mixed solution of diethyl ether (20 mL) and methanol (2 mL), and 2M (E)-1-methyl-2-(trimethylsilyl)diazene (3.6 mL, 7.2 mmol) was slowly added dropwise at 0°C. The reaction solution was stirred at room temperature for 1.5 hours.

[0143] LCMS monitoring of the reaction completion, the reaction solution was added water (30 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by preparative (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 30% to 60%) to give (E)-2-(2-amino-4-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridin-3-yl)vinyl)-5-fluorophenyl)propan-2-ol 6 (40 mg, yield: 31%, NMR showed that the product contained about 25% of cis isomer).

[0144] 1H NMR (300 MHz, CDC13) δ 7.94 (s, 1H), 7.00 (s, 1H), 3.90 (s, 3H).

[0145] Step 2: Preparation of 2-(2-amino-4-bromo-5-chlorophenyl)propan-2-ol (7b)

[0146] Methyl 2-amino-4-bromo-5-chlorobenzoate 7a (700 mg, 2.66 mmol) was dissolved in tetrahydrofuran (10 mL) and cooled to -78 °C, then slowly added 3M methyl magnesium bromide tetrahydrofuran solution (4.4 mL, 13.3 mmol). The reaction was raised to 0 °C and stirred for 3 hours under nitrogen protection.

[0147] TLC detected that the reaction was completed, the reaction was slowly added saturated aqueous ammonium chloride solution (30 mL) at 0 °C, extracted with ethyl acetate (30 mL x 3), washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate. Concentrated under reduced pressure, the crude product was purified by silica gel column chromatography (PE:EA = 5:1) to obtain 2-(2-amino-4-bromo-5-chlorophenyl)propan-2-ol 7b (550 mg, yellow solid, yield: 78%).

[0148] 1 H NMR (300 MHz, CDC13) δ 7.09 (s, 1H), 6.83 (s, 1H), 1.59 (s, 3H).

[0149] Step 3: Preparation of ((E)-2-(2-amino-4-(2-amino-6-(2-hydroxypropyl-2-yl)pyridin-3- yl)vinyl)-5-chlorophenyl)propan-2-ol (7)

[0150] 2-(2-amino-4-bromo-5-chlorophenyl)propan-2-ol 7b (320 mg, 1.22 mmol), 2-(3-amino-5-vinylpyridin-2-yl)propan-2-ol 1b (217 mg, 1.22 mmol), palladium (II) acetate (27 mg, 0.12 mmol) and potassium carbonate (421 mg, 3.05 mmol) were dissolved in 1,4-dioxane (10 mL). The reaction was stirred at 130 °C for 16 hours under nitrogen protection.

[0151] LCMS monitoring of the reaction completion, water (30 mL) was added to the reaction solution and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by preparation (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 10% to 60%) to obtain ((E)-2-(2-amino-4-(2-amino-6-(2-hydroxypropyl-2-yl)pyridine-3- ethenyl)-5-chlorophenyl)propan-2-ol (57 mg, yellowish solid, yield: 12%).

[0152] MS (ESI): m / z 362.1 [M+H] + .

[0153] 1 H NMR (400 MHz, DMSO-d6) δ 7.82 (d, J = 1.6 Hz, 1H), 7.24 (d, J = 16.4 Hz, 1H), 7.21 (d, J = 1.6 Hz, 1H), 7.05 (d, J = 20.4 Hz, 2H), 6.93 (d, J = 16.4 Hz, 1H), 5.63 (s, 2H), 5.54 (s, 2H), 5.46 (s, 1H), 5.34 (s, 1H), 1.49 (s, 12H).

[0154] [According to Rule 26 Corrected 27.10.2025] Example 8: Preparation of compound (E)-2-(2-amino-4-(2-(5-amino-6-(2-hydroxypropan-2-yl)pyridine-3-yl)ethenyl)-5- fluorophenyl)propan-2-ol (8)

[0155] Step 1: Preparation of (E)-4,4'-(ethene-1,2-diyl)dimethyl bis(2-aminobenzoate) (8a)

[0156] Methyl 2-amino-4-bromobenzoate (500 mg, 2.17 mmol), methyl 2-amino-4-vinylbenzoate 4a (462 mg, 2.61 mmol), potassium carbonate (901 mg, 6.52 mmol), and palladium acetate (48.7 mg, 0.217 mmol) were dissolved in 1,4-dioxane (20 mL) and replaced with nitrogen three times. The reaction solution was stirred at 130°C for 16 hours.

[0157] LCMS monitoring of the reaction completion, concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1 to 1:2) to obtain (E)-4,4'-(ethene-1,2-diyl)dimethyl bis(2-aminobenzoate) 8a (500 mg, yield: 70.6%).

[0158] MS (ESI): m / z 327.2 [M+H] + .

[0159] Step 2: Preparation of (E)-2,2'-(ethene-1,2-diylbis(2-amino-4,1- phenylene))bis(propan-2-ol) (8)

[0160] (E)-4,4'-(ethene-1,2-diyl)dimethyl bis(2-aminobenzoate) 8a (500 mg, 1.53 mmol) was dissolved in tetrahydrofuran (20 mL), cooled to -78 °C, 3.0 M methyl magnesium bromide solution (15.3 mL, 46.0 mmol) was added dropwise, and stirred slowly to room temperature for 12 hours.

[0161] LCMS showed the reaction was completed, saturated ammonium chloride (20 mL) was added to the reaction solution and extracted with ethyl acetate (40 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 preparation (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 0% to 100%) to give (E)-2,2'-(ethene-1,2-diylbis(2-amino-4,1- phenylene))bis(propan-2-ol) 8 (340 mg, yield: 68%).

[0162] MS (ESI): m / z 309.1 [M-H2O+H] + .

[0163] 1 H NMR (400 MHz, DMSO-d6) d 7.99 (d, J = 8.0 Hz, 2H), 6.86 (s, 2H), 6.79 (d, J = 5.6 Hz, 2H), 6.68 (dd, J1= 1.6 Hz, J2= 8.0 Hz, 2H), 5.42 (br s, 4H), 5.19 (s, 2H), 1.49 (s, 12H).

[0164] [Amended in accordance with Rule 26 27.10.2025] Example 9: Preparation of compound (E)-2-(2-amino-4-(4-amino-3-(2-hydroxypropan-2-yl)styryl)phenyl)propan-2-ol (9)

[0165] Step 1: Preparation of methyl 2-amino-5-vinylbenzoate (9a)

[0166] Methyl 2-amino-5-bromobenzoate (1.5 g, 6.52 mmol), potassium vinyltrifluoroborate (1.3 g, 9.78 mmol), 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (358 mg, 0.489 mmol) and triethylamine (3.6 mL, 26.1 mmol) were dissolved in ethanol (20 mL) and purged with nitrogen three times. The reaction was stirred at 90 °C for 5 h.

[0167] LCMS indicated the reaction was complete and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 15:1 to 3:1) to give methyl 2-amino-5-vinylbenzoate 9a (900 mg, yellow oil, yield: 78.0%).

[0168] MS (ESI): m / z 178.2 [M+H] + .

[0169] Step 2: Preparation of (E) methyl 2-amino-4-(4-amino-3-(methoxycarbonyl)styryl)benzoate (9b)

[0170] Methyl 2-amino-4-bromobenzoate (550 mg, 2.39 mmol), methyl 2-amino-5-vinylbenzoate 9a (550 mg, 3.11 mmol), potassium carbonate (991 mg, 7.17 mmol) and palladium acetate (53.7 mg, 0.239 mmol) were dissolved in 1,4-dioxane (20 mL) and purged with nitrogen three times. The reaction was stirred at 120 °C for 16 h.

[0171] LCMS indicated the reaction was complete and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 5:1 to 3:1) to give (E) methyl 2-amino-4-(4-amino-3-(methoxycarbonyl)styryl)benzoate 9b (500 mg, yield: 64.1%).

[0172] MS (ESI): m / z 327.1 [M+H] + .

[0173] Step 3: Preparation of (E)-2-(2-amino-4-(4-amino-3-(2-hydroxypropan-2-yl)styryl)phenyl)propan-2-ol (9)

[0174] (E) methyl 2-amino-4-(4-amino-3-(methoxycarbonyl)styryl)benzoate 9b (300 mg, 0.919 mmol) was dissolved in tetrahydrofuran (20 mL) and cooled to -78 °C. 3.0 M methyl magnesium bromide solution (7.7 mL, 23.0 mmol) was added dropwise and the reaction was allowed to warm to room temperature and stirred for 12 h.

[0175] LCMS monitoring of the reaction completion, saturated ammonium chloride (20 mL) was added to the reaction solution, and extracted with ethyl acetate (40 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 preparation (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 0% to 100%) to obtain (E)-2-(2-amino-4-(4-amino-3-(2-hydroxypropan-2-yl)styryl)phenyl)propan-2-ol 9 (161 mg, yield: 53.7%).

[0176] MS (ESI): m / z 309.1 [M-H2O+H] + .

[0177] 1 H NMR (400 MHz, DMSO-d6) δ 7.19 (m, 2H), 6.96 (d, J = 8.0 Hz, 1H), 6.88 (d, J = 16.0 Hz, 1H), 6.72 (m, 2H), 6.64 (dd, J1= 1.6 Hz, J2= 8.0 Hz, 1H), 6.60 (d, J = 8.4 Hz, 1H), 5.60 (br s, 2H), 5.36 (br s, 2H), 5.23 (s, 1H), 5.16 (s, 1H), 1.53 (s, 6H), 1.49 (s, 6H).

[0178] [According to Rule 26 Correction 27.10.2025] Example 10: Preparation of compound 2-(3-amino-5-((5-amino-6-(2-hydroxypropan-2-yl)pyridin-2-yl)ethynyl)pyridin-2-yl)propan-2-ol (10)

[0179] Step 1: Preparation of methyl 3-amino-5-((trimethylsilyl)ethynyl)pyridine- carboxylate (10a)

[0180] Under nitrogen protection, to a solution of methyl 3-amino-5-bromopyridine- carboxylate (1.2 g, 5.19 mmol), cuprous iodide (98.8 mg, 0.519 mmol) and bis(triphenylphosphine)palladium dichloride (364 mg, 0.519 mmol) in 1,4-dioxane (30 mL) were added ethyltrimethylsilane (1.53 g, 15.6 mmol) and triethylamine (3.6 mL, 25.96 mmol) successively. The reaction solution was stirred at 90 °C for 16 hours.

[0181] LCMS monitoring of the reaction completion, saturated ammonium chloride (20 mL) was added to the reaction solution, and extracted with ethyl acetate (40 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 preparation (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 0% to 100%) to obtain (E)-2-(2-amino-4-(4-amino-3-(2-hydroxypropan-2-yl)styryl)phenyl)propan-2-ol 9 (161 mg, yield: 53.7%).

[0182] MS (ESI): m / z 249.2 [M+H] + .

[0183] Step 2: Preparation of methyl 3-amino-5-ethynylpyridinecarboxylate (10b)

[0184] To a solution of methyl 3-amino-5-((trimethylsilyl)ethynyl)pyridinecarboxylate 10a (1.5 g, 6.04 mmol) in methanol (20 mL), potassium carbonate (1.67 g, 12.08 mmol) was added. The reaction was stirred at room temperature for 1 h.

[0185] LCMS showed the reaction was completed, water (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 silica gel column chromatography (PE:EA = 5:1 to 3:1) to give methyl 3-amino-5-ethynylpyridinecarboxylate 10b (900 mg, yellow solid, yield: 84.8%).

[0186] MS (ESI): m / z 177.1 [M+H] + .

[0187] Step 3: Preparation of methyl 3-amino-5-((5-amino-6-(methoxycarbonyl)pyridin-2- yl)ethynyl)pyridinecarboxylate (10c)

[0188] Methyl 3-amino-5-bromopyridinecarboxylate (650 mg, 2.81 mmol), methyl 3-amino-5-ethynylpyridinecarboxylate 10b (644 mg, 3.66 mmol), cuprous iodide (53.5 mg, 0.281 mmol), dichlorobis(triphenylphosphine)palladium (197 mg, 0.281 mmol) and triethylamine (3.6 mL, 25.96 mmol) were dissolved in 1,4-dioxane (20 mL) and replaced with nitrogen three times. The reaction was stirred at 95 °C for 16 h.

[0189] LCMS showed the reaction was completed, the reaction was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 3:1 to 1:1) to give methyl 3-amino-5-((5-amino-6-(methoxycarbonyl)pyridin-2-yl)ethynyl)pyridinecarboxylate 10c (620 mg, yellow solid, yield: 67.6%).

[0190] MS (ESI): m / z 327.2 [M+H] + .

[0191] Step 4: Preparation of synthesis of 2-(3-amino-5-((5-amino-6-(2-hydroxypropan-2- yl)pyridin-2-yl)ethynyl)pyridin-2-yl)propan-2-ol (10)

[0192] Methyl 3-amino-5-((5-amino-6-(methoxycarbonyl)pyridin-2-yl)ethynyl)picolinate 10c (300 mg, 0.92 mmol) was dissolved in tetrahydrofuran (12 mL) and cooled to -70 °C, 3.0 M methyl magnesium bromide solution (6.1 mL, 18.4 mmol) was added dropwise and stirred slowly to room temperature for 12 h.

[0193] 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% aqueous ammonium bicarbonate solution = 0% to 100%) to give 2-(3-amino-5-((5-amino-6-(2-hydroxypropan-2-yl)pyridin-2-yl)ethynyl)pyridin-2-yl)propan-2-ol 10 (32 mg, white solid, yield: 10.7%).

[0194] MS (ESI): m / z 327.1 [M+H] + .

[0195] 1 H NMR (400 MHz, DMSO-d6) d 7.81 (d, J = 2.0 Hz, 1H), 7.21 (d, J = 8.0 Hz, 1H), 7.06 (d, J = 2.0 Hz, 1H), 6.94 (d, J = 8.4 Hz, 1H), 5.99 (br s, 2H), 5.69 (br s, 2H), 5.54 (s, 1H), 5.51 (s, 1H), 1.49 (s, 6H), 1.48 (s, 6H).

[0196] [Amended in accordance with Rule 26 - Corrected 27.10.2025] Example 11: Preparation of compound 2-(2-amino-4-((5-amino-6-(2-hydroxypropan-2-yl)pyridin-3-yl)ethynyl)phenyl)propan-2-ol (11)

[0197] Step 1: Preparation of methyl 2-amino-4-((trimethylsilyl)ethynyl)benzoate (11a)

[0198] To a solution of methyl 2-amino-4-bromobenzoate (1.2 g, 5.22 mmol), cuprous iodide (99.4 mg, 0.522 mmol), bis(triphenylphosphine)palladium dichloride (366 mg, 0.522 mmol) in 1,4-dioxane (30 mL) was added ethyltrimethylsilane (1.54 g, 15.65 mmol) and triethylamine (3.6 mL, 26.1 mmol) successively under nitrogen protection. The reaction was stirred at 90 °C for 5 h.

[0199] LCMS showed the reaction was completed, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 15:1) to give methyl 2-amino-4-((trimethylsilyl)ethynyl)benzoate 11a (1.2 g, brown oil, yield: 93%).

[0200] MS (ESI): m / z 248.2 [M+H] + .

[0201] Step 2: Preparation of methyl 2-amino-4-ethynylbenzoate (11b)

[0202] To a solution of methyl 2-amino-4-((trimethylsilyl)ethynyl)benzoate 11a (1.2 g, 4.85 mmol) in methanol (15 mL) was added potassium carbonate (1.34 g, 9.7 mmol). The reaction was stirred at room temperature for 0.5 h.

[0203] LCMS showed the reaction was completed, and water (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 silica gel column chromatography (PE:EA = 10:1) to give methyl 2-amino-4-ethynylbenzoate 11b (750 mg, white solid, yield: 88.2%).

[0204] MS (ESI): m / z 176.1 [M+H] + .

[0205] Step 3: Preparation of methyl 3-amino-5-((3-amino-4-(methoxycarbonyl)phenyl)ethynyl)picolinate (11c)

[0206] Methyl 3-amino-5-bromopicolinate (500 mg, 2.16 mmol), methyl 2-amino-4- ethynylbenzoate 11b (455 mg, 2.6 mmol), cuprous iodide (41 mg, 0.216 mmol), bis(triphenylphosphine)palladium dichloride (152 mg, 0.216 mmol) and triethylamine (1.5 mL, 10.8 mmol) were dissolved in 1,4-dioxane (20 mL) and purged with nitrogen for three times. The reaction was stirred at 95 °C for 16 h.

[0207] LCMS showed the reaction was completed and the reaction was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 4:1 to 1:1) to give methyl 3-amino-5-((3-amino-4- (methoxycarbonyl)phenyl)ethynyl)picolinate 11c (500 mg, yellow solid, yield: 71.1%).

[0208] MS (ESI): m / z 326.2 [M+H] + .

[0209] Step 4: Preparation of 2-(2-amino-4-((5-amino-6-(2-hydroxypropan-2-yl)pyridin-3- yl)ethynyl)phenyl)propan-2-ol (11)

[0210] Methyl 3-amino-5-((3-amino-4-(methoxycarbonyl)phenyl)ethynyl)picolinate 11c (500 mg, 1.54 mmol) was dissolved in tetrahydrofuran (20 mL) and cooled to -78 °C. 3.0 M methyl magnesium bromide solution (10.2 mL, 30.7 mmol) was added dropwise and the reaction was stirred at room temperature for 12 h.

[0211] LCMS showed the reaction was completed and saturated ammonium chloride (20 mL) was added to the reaction and extracted with ethyl acetate (40 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 preparative (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 0% to 100%) to give 2-(2-amino-4-((5-amino-6-(2-hydroxypropan-2-yl)pyridin-3- yl)ethynyl)phenyl)propan-2-ol (75 mg, white solid, yield: 15%).

[0212] MS (ESI): m / z 326.1 [M+H] + .

[0213] 1H NMR (400 MHz, DMSO-d6) δ 7.80 (d, J = 2.0 Hz, 1H), 7.05 (d, J = 2.0 Hz, 1H), 7.06 (d, J = 8.0 Hz, 1H), 6.78 (d, J = 2.0 Hz, 1H), 6.65 (dd, Ji = 1.6 Hz, J2= 8.0 Hz, 1H), 5.69 (br s, 2H), 5.58 (br s, 2H), 5.51 (s, 1H), 5.30 (s, 1H), 1.49 (s, 6H), 1.48 (s, 6H).

[0214] [According to Rule 26 Correction 27.10.2025] Example 12: Preparation of compound 2-(2-amino-4-((4-amino-3-(2-hydroxypropan-2-yl)phenyl)ethynyl)phenyl)propan-2-ol (12)

[0215] Step 1: Preparation of methyl 4-((3-acetyl-4-aminophenyl)ethynyl)-2- aminobenzoate (12a)

[0216] Methyl 2-amino-4-ethynylbenzoate 11b (436 mg, 2.49 mmol), copper iodide (36.6 mg, 0.192 mmol), palladium dichloride bis(triphenylphosphine) (135 mg, 0.192 mmol) and triethylamine (1.38 mL, 9.96 mmol) were dissolved in 1,4-dioxane (20 mL) and replaced with nitrogen three times. The reaction was stirred at 50 °C for 16 hours.

[0217] LCMS showed the reaction was completed and the reaction was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 5:1 to 3:1) to give methyl 4-((3-acetyl-4-aminophenyl)ethynyl)-2-aminobenzoate 12a (400 mg, yellow solid, yield: 65.1%).

[0218] MS (ESI): m / z 309.1 [M+H] + .

[0219] Step 2: Preparation of 2-(2-amino-4-((4-amino-3-(2-hydroxypropan-2-yl)phenyl)ethynyl)phenyl)propan-2-ol (12)

[0220] Methyl 4-((3-acetyl-4-aminophenyl)ethynyl)-2-aminobenzoate 12a (400 mg, 1.3 mmol) was dissolved in tetrahydrofuran (20 mL) and cooled to -78 °C. A 3.0 M solution of methylmagnesium bromide (8.6 mL, 25.9 mmol) was added dropwise and the reaction was allowed to warm to room temperature slowly over 12 h.

[0221] LCMS indicated the reaction was complete, saturated ammonium chloride (20 mL) was added to the reaction and extracted with ethyl acetate (40 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% aqueous ammonium bicarbonate = 0% to 100%) to give 2-(2-amino-4-((4-amino-3-(2-hydroxypropan-2-yl)phenyl)ethynyl)phenyl 12 (230 mg, white solid, yield: 54.6%).

[0222] MS (ESI): m / z 307.1 [M-H2O+H] + .

[0223] 1 H NMR (400 MHz, DMSO-d6) δ 7.11 (d, J = 1.6 Hz, 1H), 7.07 (dd, J1= 2.0 Hz, J2= 8.4 Hz, 1H), 6.98 (d, J = 8.0 Hz, 1H), 6.71 (d, J = 1.6 Hz, 1H), 6.59 (m, 2H), 5.81 (br s, 2H), 5.50 (br s, 2H), 5.30 (s, 1H), 5.25 (s, 1H), 1.50 (s, 6H), 1.48 (s, 6H).

[0224] [Amended according to Rule 26 - Corrected 27.10.2025] Example 13: Preparation of compound 2,2'-(ethyn-1,2-diylbis(2-amino-4,1-phenylene))bis(propan-2-ol) (13)

[0225] Step 1: Preparation of 4,4'-(ethyn-1,2-diyl)bis(2-aminobenzoic acid dimethyl ester) (13a)

[0226] Methyl 2-amino-4-bromobenzoate (500 mg, 2.17 mmol), methyl 2-amino-4-ethynylbenzoate 11b (457 mg, 2.61 mmol), cuprous iodide (41.3 mg, 0.217 mmol), palladium dichloride bis(triphenylphosphine) (152 mg, 0.217 mmol) and triethylamine (1.51 mL, 10.86 mmol) were dissolved in 1,4-dioxane (20 mL) and purged with nitrogen three times. The reaction was stirred at 95 °C for 16 h.

[0227] LCMS detected the completion of the reaction, the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1 to 4:1) to give 4,4'-(ethyn-l,2-diyl)bis(2- aminobenzoic acid dimethyl ester 13a (480 mg, yellow solid, yield: 68.2%).

[0228] MS (ESI): m / z 325.1 [M+H] + .

[0229] Step 2: Preparation of 2,2'-(ethyn-l,2-diylbis(2-amino-4,l-phenylene))bis(propan-2-ol) (13)

[0230] 4,4'-(ethyn-l,2-diyl)bis(2-aminobenzoic acid dimethyl ester (480 mg, 1.48 mmol) was dissolved in tetrahydrofuran (20 mL) and cooled to -78 °C, 3.0 M methyl magnesium bromide solution (9.9 mL, 29.6 mmol) was added dropwise, and stirred slowly to room temperature for 12 hours.

[0231] LCMS detected the completion of the reaction, the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1 to 4:1) to give 4,4'-(ethyn-l,2-diyl)bis(2- aminobenzoic acid dimethyl ester 13a (480 mg, yellow solid, yield: 68.2%).

[0232] MS (ESI): m / z 325.1 [M+H] + .

[0233] 1 H NMR (400 MHz, DMSO-d6) d 7.01 (d, J = 8.0 Hz, 2H), 6.75 (d, J = 2.0 Hz, 2H), 6.62 (dd, Ji = 2.0 Hz, J2= 8.0 Hz, 2H), 5.55 (br s, 4H), 5.28 (s, 2H), 1.49 (s, 12H).

[0234] [Amended in accordance with Rule 26 27.10.2025] Example 14: Preparation of compound 2,2'-(ethen-l,l-diylbis(2-amino-4,l-phenylene))bis(propan-2-ol) (14)

[0235] Step 1: Preparation of methyl 2-nitro-4-vinylbenzoate (14a)

[0236] Methyl 4-bromo-2-nitrobenzoate (2.0 g, 7.69 mmol), potassium vinyltrifluoroborate (1.75 g, 13.08 mmol), 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (422 mg, 0.577 mmol) and triethylamine (5.3 mL, 38.45 mmol) were dissolved in ethanol (25 mL) and purged with nitrogen three times. The reaction was stirred at 95 °C for 4 h.

[0237] LCMS showed the reaction was completed and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 12:1) to give methyl 2-nitro-4-vinylbenzoate 14a (1.5 g, yellow oil, yield: 94.6%).

[0238] MS (ESI): m / z 208.1 [M+H] + .

[0239] Step 2: Preparation of methyl 4-(1,2-dibromoethyl)-2-nitrobenzoate (14b)

[0240] To a solution of methyl 2-nitro-4-vinylbenzoate 14a (1.5 g, 7.24 mmol) in 1,2-dichloroethane (30 mL) was added bromine (3.47 g, 21.7 mmol) at room temperature. The reaction was stirred at room temperature for 1 h.

[0241] LCMS showed the reaction was completed and the reaction was diluted with dichloromethane (150 mL), washed with saturated sodium bisulfite (40 mL x 3), saturated sodium chloride (40 mL x 3), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give methyl 4-(1,2-dibromoethyl)-2-nitrobenzoate 14b (2.6 g, yellow oil, yield: 97.9%).

[0242] MS (ESI): m / z 367.9 [M+H] + .

[0243] 1 H NMR (400 MHz, DMSO-d6) d 8.16 (d, J = 2.0 Hz, 1H), 7.94 (dd, J1= 2.4 Hz, J2= 10.8 Hz, 1H), 7.87 (d, J = 10.8 Hz, 1H), 5.53 (dd, J1= 3.2 Hz, J2= 10.0 Hz, 1H), 4.33-4.20 (m, 2H), 3.95 (s, 3H).

[0244] Step 3: Preparation of methyl 4-(1-bromo vinyl)-2-nitrobenzoate (14c)

[0245] To a solution of methyl 4-(1,2-dibromoethyl)-2-nitrobenzoate 14b (2.6 g, 7.085 mmol) in tetrahydrofuran (15 mL) and methanol (15 mL) was added potassium carbonate (2.94 g, 21.25 mmol). The reaction was stirred at room temperature for 30 min.

[0246] The reaction was monitored by LCMS and upon completion, the reaction was diluted with ethyl acetate (200 mL), washed with saturated sodium chloride (40 mL x 3), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 12:1) to give methyl 4-(1-bromo vinyl)-2-nitrobenzoate 14c (1.9 g, yellow oil, yield: 93.6%).

[0247] MS (ESI): m / z 287.9 [M+2+H] + .

[0248] 1 H NMR (400 MHz, DMSO-d6) δ 8.28 (d, J = 2.0 Hz, 1H), 8.09 (dd, Ji = 2.0 Hz, J2= 8.0 Hz, 1H), 7.95 (d, J = 8.4 Hz, 1H), 6.76 (d, J = 3.2 Hz, 1H), 6.16 (d, J = 3.2 Hz, 1H), 3.87 (s, 3H).

[0249] Step 4: Preparation of methyl 2-amino-4-(1-(4-(methoxycarbonyl)-3-nitrophenyl)vinyl)benzoate (14d)

[0250] To a solution of methyl 4-(1-bromo vinyl)-2-nitrobenzoate 14c (900 mg, 3.14 mmol), methyl 2-amino-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (1.3 g, 4.72 mmol), potassium carbonate (1.3 g, 9.43 mmol) and 1,1 '-bis(diphenylphosphino)ferrocene palladium(II) dichloride in water (1.0 mL) and 1,4-dioxane (15 mL) was purged with nitrogen three times. The reaction was stirred at 95 °C for 4 h.

[0251] The reaction was monitored by LCMS and upon completion, the reaction was cooled to room temperature, diluted with ethyl acetate (150 mL), washed with saturated sodium chloride (40 mL x 3), 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 7:1) to give methyl 2-amino-4-(1-(4-(methoxycarbonyl)-3-nitrophenyl)vinyl)benzoate 14d (430 mg, yellow oil, yield: 38.4%).

[0252] MS (ESI): m / z 357.1 [M+H] + .

[0253] Step 5: Preparation of methyl 2-amino-4-(l-(4-(methoxycarbonyl)-3- aminophenyl)vinyl)benzoate (14e)

[0254] To a solution of methyl 2-amino-4-(l-(4-(methoxycarbonyl)-3-nitrophenyl)vinyl)benzoate 14d (430 mg, 1.2 mmol) in water (5.0 mL) and ethanol (20 mL) was added iron powder (337 mg, 6.03 mmol) and ammonium chloride (323 mg, 6.03 mmol) successively. The reaction was stirred at 85 °C for 1 h.

[0255] LCMS showed the reaction was completed. The reaction was cooled to room temperature, diluted with ethyl acetate (150 mL), washed with saturated sodium chloride solution (30 mL x 3), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 8:1 to 4:1) to give methyl 2-amino-4-(l-(4-(methoxycarbonyl)-3- aminophenyl)vinyl)benzoate 14e (340 mg, yellow oil, yield: 86.3%).

[0256] MS (ESI): m / z 327.1 [M+H] + .

[0257] Step 6: Preparation of 2,2'-(ethene-1,1-diylbis(2-amino-4,1- phenylene))bis(propan-2-ol) (14)

[0258] Methyl 2-amino-4-(l-(4-(methoxycarbonyl)-3-aminophenyl)vinyl)benzoate 14e (340 mg, 1.04 mmol) was dissolved in tetrahydrofuran (20 mL) and cooled to -78 °C. 3.0 M methyl magnesium bromide solution (10.4 mL, 31.3 mmol) was added dropwise and the reaction was stirred at room temperature for 16 h.

[0259] LCMS showed the reaction was completed. Saturated ammonium chloride solution (50 mL) was added to the reaction and extracted with ethyl acetate (40 mL x 3). The organic phase was washed with saturated sodium chloride solution (30 mL x 3), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by preparative (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 0% to 100%) to give 2,2'-(ethene-1,1-diylbis(2-amino-4,1- phenylene))bis(propan-2-ol) 14 (70 mg, white solid, yield: 20.6%).

[0260] MS (ESI): m / z 309.1 [M-H20+H] + .

[0261] 1 H NMR (400 MHz, DMSO-d6) δ 6.98 (d, J = 8.0 Hz, 2H), 6.55 (d, J = 1.6 Hz, 2H), 6.42 (dd, Ji = 1.6 Hz, J2= 10.4 Hz, 2H), 5.41 (br s, 4H), 5.22 (s, 2H), 5.18 (s, 2H), 1.50 (s, 12H).

[0262] Example 15: Preparation of compound (E)-2,2'-(ethene-1,2-diylbis(2-amino-5,1- phenylene))bis(propan-2-ol) (15)

[0263] Example 16: Preparation of compound 2,2'-(ethene-1,1-diylbis(2-amino-1,1- phenylene))bis(propan-2-ol) (16)

[0264] Synthetic route of compounds 15 and 16:

[0265] Step 1: Preparation of (E)-dimethyl 5,5'-(ethene-1,2-diyl)bis(2-aminobenzoate) (15a) and 5,5'-(ethene-1,1-diyl)dimethyl bis(2-aminobenzoate) (16a)

[0266] Methyl 2-amino-5-bromobenzoate (1.12 g, 4.86 mmol), methyl 2-amino-5- vinylbenzoate 9a (1.08 g, 6.08 mmol), potassium carbonate (2.0 g, 14.6 mmol) and palladium acetate (109 mg, 0.486 mmol) were dissolved in 1,4-dioxane (30 mL) and purged with nitrogen three times. The reaction was stirred at 120 °C for 16 h.

[0267] LCMS indicated the reaction was complete and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 5:1 to 3:1) to give a mixture of (E)-dimethyl 5,5'-(ethene-1,2-diyl)bis(2-aminobenzoate) 15a and 5,5'-(ethene-1,1-diyl)dimethyl bis(2-aminobenzoate) 16a (700 mg, yellow solid, yield: 44.1%).

[0268] MS (ESI): m / z 327.1 [M+H] + .

[0269] Step 2: Preparation of (E)-2,2'-(ethene-1,2-diylbis(2-amino-5,1- phenylene))bis(propan-2-ol) (15) and 2,2'-(ethene-1,1-diylbis(2-amino-1,1- phenylene))bis(propan-2-ol) (16)

[0270] A mixture of (E)-dimethyl 5,5'-(ethene-1,2-diyl)bis(2-aminobenzoate) 15a and 5,5'-(ethene-1,1-diyl)dimethyl bis(2-aminobenzoate) 16a (500 mg, 1.53 mmol) was dissolved in tetrahydrofuran (60 mL), cooled to -78 °C, 3.0 M methylmagnesium bromide solution (12.8 mL, 38.3 mmol) was added dropwise, and stirred slowly to room temperature for 16 hours.

[0271] LCMS indicated the reaction was completed, saturated ammonium chloride (20 mL) was added to the reaction, and extracted with ethyl acetate (50 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 preparation (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 0% to 100%) to give (E)-2,2'-(ethene-1,2-diylbis(2-amino-5,1- phenylene))bis(propan-2-ol) 15 (168 mg, white solid-like) and 2,2'-(ethene-1,1-diylbis(2-amino-1,1-phenylene))bis(propan-2-ol) 16 (21 mg, white solid-like, total yield: 36.4%).

[0272] The mass spectrum and nuclear magnetic resonance hydrogen spectrum of compound 15 are as follows:

[0273] MS (ESI): m / z 327.1 [M+H] + .

[0274] 1 H NMR (400 MHz, DMSO-d6) δ 7.14-7.12 (m, 4H), 6.74 (s, 2H), 6.57 (d, J = 7.6 Hz, 2H), 5.50 (br s, 4H), 5.21 (s, 2H), 1.53 (s, 12H).

[0275] The mass spectrum and nuclear magnetic resonance hydrogen spectrum of compound 16 are as follows:

[0276] MS (ESI): m / z 327.1 [M+H] + .

[0277] 1H NMR (400 MHz, DMSO-d6) δ 6.98 (d, J = 1.6 Hz, 2H), 6.91 (dd, Ji = 2.0 Hz, J2= 8.4 Hz, 2H), 6.57 (d, J = 7.6 Hz, 2H), 5.52 (br s, 4H), 5.22 (s, 2H), 5.20 (s, 2H), 1.45 (s, 12H).

[0278] [According to Rule 26 Correction 27.10.2025] Example 17: Preparation of compound 2-(2-amino-4-(2-(4-amino-3-(2-hydroxypropan-2-yl)phenoxy)ethoxy)phenyl)propan-2-ol (17)

[0279] Step 1: Preparation of methyl 5-hydroxy-2-nitrobenzoate (17a)

[0280] Methyl 5-hydroxy-2-nitrobenzoate (6.0 g, 32.8 mmol) was dissolved in methanol (60 mL), then concentrated sulfuric acid (1 mL) was added dropwise. It was stirred at 80 °C for 12 h under nitrogen protection.

[0281] LCMS showed the reaction was completed, concentrated under reduced pressure, the crude product was purified by silica gel column chromatography (PE:EA:Et3N = 5:1:0.1% to 1:3:0.1%) to give methyl 5-hydroxy-2-nitrobenzoate 17a (5.2 g, yellow solid, yield: 80%).

[0282] MS (ESI): m / z 215.20 [M+NH4] + .

[0283] Step 2: Preparation of methyl 5-(2-bromoethoxy)-2-nitrobenzoate (17b) and methyl 5,5'-(ethane-1,2-diylbis(oxy))bis(2-nitrobenzoate) (17b-1)

[0284] Methyl 5-hydroxy-2-nitrobenzoate 17a (1.2 g, 6.09 mmol) was dissolved in N,N- dimethylformamide (12 mL), 1,2-dibromoethane (1.72 g, 9.14 mmol) and potassium carbonate (1.68 g, 12.2 mmol) were added. The reaction was stirred at 80 °C for 12 h under nitrogen protection.

[0285] LCMS showed the reaction was completed, water (20 mL) was added to the reaction solution 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, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA:Et3N=5:1:0.1% to 1:3:0.1%) to give 5-(2-bromoethoxy)-2-nitrobenzoic acid methyl ester (900 mg, yellow oil, yield: 49%) 17b and 5,5'-(ethane-1,2-diylbis(oxy))bis(2-nitrobenzoic acid methyl ester) (220 mg, yellow solid, yield: 17%) 17b-1.

[0286] The mass spectrum and nuclear magnetic resonance hydrogen spectrum of compound 17b are as follows:

[0287] MS (ESI): m / z 321.0 [M+NH4] + .

[0288] 1 H NMR (300 MHz, CDCl3) δ 8.04 (d, J = 8.7 Hz, 1H), 7.09-7.03 (m, 2H), 4.39 (t, J = 6.0 Hz, 2H), 3.94 (s, 3H), 3.67 (t, J = 6.0 Hz, 2H).

[0289] The mass spectrum and nuclear magnetic resonance hydrogen spectrum of compound 17b-1 are as follows:

[0290] MS (ESI): m / z 438.20 [M+NH4] + .

[0291] 1 H NMR (300 MHz, CDCl3) δ 8.05 (d, J = 9.0 Hz, 2H), 7.13-7.07 (m, 4H), 4.46 (s, 4H), 3.94 (s, 6H).

[0292] Step 3: Preparation of methyl 4-(2-(3-(methoxycarbonyl)-4-nitrophenoxy)ethoxy)-2- nitrobenzoate (17c)

[0293] Methyl 5-(2-bromoethoxy)-2-nitrobenzoate 17b (500 mg, 1.64 mmol) was dissolved in N,N-dimethylformamide (5 mL), and methyl 4-hydroxy-2-nitrobenzoate (323 mg, 1.64 mmol) and potassium carbonate (453 mg, 3.28 mmol) were added. The reaction solution was stirred at 80°C under nitrogen protection for 12 hours.

[0294] LCMS monitoring of the reaction completion, water (20 mL) was added to the reaction solution 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, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA:Et3N = 5:1:0.1% to 1:3:0.1%) to obtain methyl 4-(2-(3-(methoxycarbonyl)-4- nitrophenoxy)ethoxy)-2-nitrobenzoate 17c (450 mg, yellow solid, yield: 65%).

[0295] MS (ESI): m / z 438.2 [M+NH4] + .

[0296] 1H NMR (300 MHz, CDC13) δ 8.05 (d, J = 8.7 Hz, 1H), 7.81 (d, J = 8.7 Hz, 1H), 7.32 (d, J = 2.4 Hz, 1H), 7.18-7.06 (m, 3H), 4.46 (s, 4H), 3.94 (s, 3H), 3.89 (s, 3H).

[0297] Step 4: Preparation of methyl 2-amino-4-(2-(4-amino-3-(methoxycarbonyl)phenoxy)ethoxy)benzoate (17d)

[0298] Methyl 4-(2-(3-(methoxycarbonyl)-4-nitrophenoxy)ethoxy)-2-nitrobenzoate 17c (450 mg, 1.07 mmol) was dissolved in methanol (5 mL), tetrahydrofuran (5 mL), and N,N-dimethylformamide (5 mL), and 10% palladium-carbon (45 mg) was added. Hydrogenation was performed with a hydrogen balloon at room temperature for 12 hours.

[0299] LCMS monitoring of the reaction completion, water (20 mL) was added to the reaction solution 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, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA:Et3N = 5:1:0.1% to 1:3:0.1%) to obtain methyl 4-(2-(3-(methoxycarbonyl)-4- nitrophenoxy)ethoxy)-2-nitrobenzoate 17c (450 mg, yellow solid, yield: 65%).

[0300] MS (ESI): m / z 361.20 [M+H] + .

[0301] 1H NMR (300 MHz, DMSO-d6) δ 7.63 (d, J = 9.0 Hz, 1H), 7.23 (d, J = 3.0 Hz, 1H), 7.03 (dd, J = 9.0 Hz, 3.0 Hz, 1H), 6.75 (d, J = 9.3 Hz, 1H), 6.69 (br s, 2H), 6.37 - 6.31 (m, 3H), 6.17 (dd, J = 9.0 Hz, 2.4 Hz, 1H), 4.26 - 4.14 (m, 4H), 3.78 (s, 3H), 3.74 (s, 3H).

[0302] Step 5: Preparation of 2-(2-amino-4-(2-(4-amino-3-(2-hydroxypropan-2- yl)phenoxy)ethoxy)phenyl)propan-2-ol (17)

[0303] Methyl 2-amino-4-(2-(4-amino-3-(methoxycarbonyl)phenoxy)ethoxy)benzoate 17d (340 mg, 0.944 mmol) was dissolved in tetrahydrofuran (3 mL) and cooled to -78 °C, 3.0 M methyl magnesium bromide solution (9.4 mL, 28.2 mmol) was added dropwise and stirred slowly to room temperature for 12 h.

[0304] LCMS indicated the reaction was completed, water (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, concentrated under reduced pressure, the crude product was purified by prep (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 20% to 50%) to give 2-(2-amino-4-(2-(4-amino-3-(2-hydroxypropan-2- yl)phenoxy)ethoxy)phenyl)propan-2-ol 17 (75 mg, light yellow solid, yield: 22%).

[0305] MS (ESI): m / z 325.1 [M-2H2O + H] + .

[0306] 1 H NMR (400 MHz, DMSO-d6) δ 6.89 (d, J = 8.4 Hz, 1H), 6.65 (d, J = 2.8 Hz, 1H), 6.62 (dd, J = 8.4 Hz, 2.8 Hz, 1H), 6.54 (d, J = 8.4 Hz, 1H), 6.21 (d, J = 2.4 Hz, 1H), 6.08 (dd, J = 8.4 Hz, 2.4 Hz, 1H), 5.42 (br s, 2H), 5.18 (br s, 1H), 5.08 (s, 1H), 5.02 (br s, 2H), 4.11 (s, 4H), 1.48 (s, 6H), 1.46 (s, 6H).

[0307] [Corrected according to Rule 26 27.10.2025] Example 18: Preparation of compound 2,2'-((ethane-1,2-diylbis(oxy))bis(6-amino-3,1- phenylene))bis(propan-2-ol) trifluoroacetate salt (18)

[0308] Step 1: Preparation of 5,5'-(ethane-1,2-diylbis(oxy))bis(2-nitrobenzoic acid methyl ester) (18a)

[0309] 5,5'-(ethane-1,2-diylbis(oxy))bis(2-nitrobenzoic acid methyl ester) 17b-1 (220 mg, 0.523 mmol) was dissolved in methanol (5 mL), tetrahydrofuran (5 mL) and ethyl acetate (5 mL), and 10% palladium on carbon (44 mg) was added. Hydrogenation was carried out with a hydrogen balloon at room temperature for 12 hours.

[0310] LCMS showed the reaction was complete, and the filtrate was concentrated under reduced pressure. The crude product was obtained as 5,5'-(ethane-1,2-diylbis(oxy))bis(2- aminobenzoic acid methyl ester) 18a (200 mg of crude product, yellow solid, yield: 100%).

[0311] MS (ESI): m / z 360.80 [M+H] + .

[0312] 1 H NMR (300 MHz, CDC13) δ 7.42 (d, J = 3.0 Hz, 2H), 7.01 (dd, J = 8.7 Hz, 2.7 Hz, 2H), 6.63 (d, J = 9.0 Hz, 2H), 4.23 (s, 4H), 3.87 (s, 6H).

[0313] Step 2: Preparation of 2,2'-((ethane-1,2-diylbis(oxy))bis(6-amino-3,1- phenylene))bis(propan-2-ol) trifluoroacetate salt (18)

[0314] 5,5'-(ethane-1,2-diylbis(oxy))bis(2-nitrobenzoic acid methyl ester) 17b-1 (220 mg, 0.523 mmol) was dissolved in methanol (5 mL), tetrahydrofuran (5 mL) and ethyl acetate (5 mL), and 10% palladium on carbon (44 mg) was added. Hydrogenation was carried out with a hydrogen balloon at room temperature for 12 hours.

[0315] LCMS showed the reaction was completed, water (20 mL) was added to the reaction solution 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, concentrated under reduced pressure, and the crude product was purified by preparation (acetonitrile: 0.1% trifluoroacetic acid aqueous solution = 5% to 35%) to obtain 2,2'-((ethane-1,2-diylbis(oxy))bis(6-amino-3,1- phenylene))bis(propan-2-ol) trifluoroacetate 18 (100 mg, off-white solid, yield: 32%).

[0316] MS (ESI): m / z 325.1 [M-2H2O + H] + .

[0317] 1 H NMR (400 MHz, DMSO-d6) δ 9.42 (br s, 4H), 7.20 (d, J = 8.8 Hz, 2H), 6.97 (dd, J = 8.4 Hz, 2.8 Hz, 2H), 6.93 (d, J = 2.4 Hz, 2H), 4.32 (s, 4H), 1.55 (s, 12H).

[0318] [Amended in accordance with Rule 26 27.10.2025] Example 19: Preparation of compound 2-(2-amino-5-(2-((5-amino-6-(2-hydroxypropan-2-yl)pyridin-3-yl)oxy)ethoxy)phenyl)propan-2-ol (19)

[0319] Step 1: Preparation of methyl 5-(2-bromoethoxy)-3-nitropyridine carboxylate (19a)

[0320] Methyl 5-hydroxy-3-nitropyridine carboxylate (3.0 g, 15.1 mmol) was dissolved in N,N-dimethylformamide (60 mL), 1,2-dibromoethane (8.54 g, 45.4 mmol) and potassium carbonate (4.17 g, 30.2 mmol) were added. The reaction solution was stirred at 80°C under nitrogen protection overnight.

[0321] LCMS showed the reaction was completed, water (20 mL) was added to the reaction solution 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, concentrated under reduced pressure, and the crude product was purified by preparation (acetonitrile: 0.1% trifluoroacetic acid aqueous solution = 5% to 35%) to obtain 2,2'-((ethane-1,2-diylbis(oxy))bis(6-amino-3,1- phenylene))bis(propan-2-ol) trifluoroacetate 18 (100 mg, off-white solid, yield: 32%).

[0322] MS (ESI): m / z 304.60 [M+H] + .

[0323] 1 H NMR (300 MHz, CDC13) δ 8.56 (d, J = 2.4 Hz, 1H), 7.70 (d, J = 2.7 Hz, 1H), 4.47 (t, J = 6.0 Hz, 2H), 3.99 (s, 3H), 3.70 (t, J = 6.0 Hz, 1H).

[0324] Step 2: Preparation of methyl 5-(2-(3-(methoxycarbonyl)-4-nitrophenoxy)ethoxy)- 3-nitropyridinecarboxylate (19b)

[0325] Methyl 5-(2-bromoethoxy)-3-nitropyridinecarboxylate (500 mg, 1.64 mmol) was dissolved in N,N-dimethylformamide (5 mL), and 5-hydroxy-2-nitrobenzoic acid methyl ester (323 mg, 1.64 mmol) and potassium carbonate (679 mg, 4.92 mmol) were added. The reaction was stirred at 80 °C under nitrogen overnight.

[0326] LCMS showed the reaction was completed, water (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, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA:Et3N = 2:1:0.1% to 1:5:0.1%) to give methyl 5-(2-(3-(methoxycarbonyl)-4-nitrophenoxy)ethoxy)-3-nitropyridinecarboxylate 19b (270 mg, yellow solid, yield: 39%).

[0327] MS (ESI): m / z 421.80 [M+H] + .

[0328] Step 3: Preparation of methyl 3-amino-5-(2-(4-amino-3-(methoxycarbonyl)phenoxy)ethoxy)pyridinecarboxylate (19c)

[0329] Methyl 5-(2-(3-(methoxycarbonyl)-4-nitrophenoxy)ethoxy)-3-nitropyridinecarboxylate 19b (270 mg, 0.641 mmol) was dissolved in N,N-dimethylformamide (5 mL) and methanol (5 mL), and 10% palladium-carbon (27 mg) was added. Hydrogenation was carried out with a hydrogen balloon at room temperature overnight.

[0330] LCMS indicated the reaction was completed, filtered with celite, concentrated under reduced pressure, the crude product was purified by silica gel column chromatography (PE:EA:Et3N=2:1:0.1% to 1:3:0.1%) to give methyl 3-amino-5-(2-(4-amino-3-(methoxycarbonyl)phenoxy)ethoxy)picolinate 19c (100 mg, yellow solid, yield: 43%).

[0331] MS (ESI): m / z 362.20 [M+H] + .

[0332] Step 4: Preparation of 2-(2-amino-5-(2-((5-amino-6-(2-hydroxypropan-2-yl)pyridin-3- yl)oxy)ethoxy)phenyl)propan-2-ol (19)

[0333] Methyl 3-amino-5-(2-(4-amino-3-(methoxycarbonyl)phenoxy)ethoxy)picolinate 19c (100 mg, 0.277 mmol) was dissolved in tetrahydrofuran (1 mL) and cooled to -78 °C, 3.0 M methyl magnesium bromide solution (5.5 mL, 16.5 mmol) was added dropwise, and the reaction was stirred slowly to room temperature for 12 hours.

[0334] LCMS indicated the reaction was completed, water (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, concentrated under reduced pressure, the crude product was purified by silica gel column chromatography (PE:EA=2:1 to 1:4, then methanol:dichloromethane=1:20) to give the crude product, which was further purified by prep (acetonitrile:0.1% aqueous ammonium bicarbonate solution=5% to 40%) to give 2-(2-amino-5-(2-((5-amino-6-(2-hydroxypropan-2-yl)pyridin-3- yl)oxy)ethoxy)phenyl)propan-2-ol 19 (18 mg, yield: 18%).

[0335] MS (ESI): m / z 362.1 [M+H] + .

[0336] 1H NMR (400 MHz, DMSO-d6) δ 7.44 (d, J = 2.8 Hz, 1H), 6.65 (d, J = 2.4 Hz, 1H), 6.62 (dd, J = 8.4 Hz, 2.8 Hz, 1H), 6.58 (d, J = 2.4 Hz, 1H), 6.54 (d, J = 8.8 Hz, 1H), 5.55 (br s, 2H), 5.32 (br s, 1H), 5.18 (br s, 1H), 5.03 (br s, 2H), 4.20-4.18 (m, 2H), 4.14-4.12 (m, 2H), 1.48 (s, 6H), 1.45 (s, 6H).

[0337] Example 20: Preparation of compound 2,2'-((ethane-1,2-diylbis(oxy))bis(2-amino-4,1- phenylene))bis(propan-2-ol) (20)

[0338] Step 1: Preparation of methyl 4-bromo-2-nitrobenzoate (20a)

[0339] Methyl 4-bromo-2-nitrobenzoate (10 g, 40.6 mmol) was dissolved in methanol (100 mL), then concentrated sulfuric acid (1 mL) was added dropwise. Stirring was carried out under nitrogen protection at 80 °C for 12 hours.

[0340] LCMS detected that the reaction was completed, and the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA:Et3N = 5:1:0.1% to 1:3:0.1%) to obtain methyl 4-bromo-2-nitrobenzoate 20a (6.4 g, yellow oil, yield: 61%).

[0341] MS (ESI): m / z 277.0 [M+NH4] + .

[0342] 1 H NMR (300 MHz, CDCl3) δ 8.02 (d, J = 1.8 Hz, 1H), 7.80 (dd, J = 8.1 Hz, 1.5 Hz, 1H), 7.65 (d, J = 8.1 Hz, 1H), 3.92 (s, 3H).

[0343] Step 2: Preparation of (4-(methoxycarbonyl)-3-nitrophenyl)boronic acid (20b)

[0344] Methyl 4-bromo-2-nitrobenzoate 20a (5.0 g, 19.2 mmol) was dissolved in 1,4-dioxane (100 mL), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (4.88 g, 19.2 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(ll) (697 mg, 0.953 mmol) and potassium acetate (2.8 g, 28.6 mmol) were added. The reaction was stirred at 80 °C under nitrogen for 12 h.

[0345] LCMS showed the reaction was completed, filtered with celite, the filtrate was concentrated under reduced pressure to give methyl 4-(methoxycarbonyl)-3-nitrophenyl)boronic acid 20b (6.5 g crude, off-white solid, yield: 100%).

[0346] MS (ESI): m / z 242.80 [M+NH4] + .

[0347] Step 3: Preparation of methyl 4-hydroxy-2-nitrobenzoate (20c)

[0348] Methyl 4-(methoxycarbonyl)-3-nitrophenyl)boronic acid 20b (6.5 g crude, 19.2 mmol) was dissolved in tetrahydrofuran (40 mL), 30% hydrogen peroxide (10.9 g, 96 mmol) was added. The reaction was stirred at room temperature under nitrogen for 12 h.

[0349] LCMS showed the reaction was completed, cooled to 0 °C with ice water, saturated sodium bisulfite aqueous solution (40 mL) was added to the cooled reaction, concentrated under reduced pressure, the crude was dissolved with methanol to give methyl 4-hydroxy-2-nitrobenzoate 20c (6.0 g crude, off-white solid, yield: 100%).

[0350] MS (ESI): m / z 214.80 [M+NH4] + .

[0351] 1 H NMR (300 MHz, DMSO-d6) δ 7.77 (d, J = 8.4 Hz, 1H), 7.23 (d, J = 2.4 Hz, 1H), 7.11 (dd, J = 8.4 Hz, 2.4 Hz, 1H), 3.93 (br s, 1H), 3.78 (s, 3H).

[0352] Step 4: Preparation of methyl 4-hydroxy-2-nitrobenzoate (20d)

[0353] Methyl 4-hydroxy-2-nitrobenzoate 20c (3 g crude, 9.60 mmol) was dissolved in methanol (30 mL), and 10% palladium on carbon (300 mg) was added. The reaction was hydrogenated with a hydrogen balloon at room temperature for 12 hours.

[0354] LCMS showed the reaction was completed, the reaction was filtered with celite, the filter cake was washed with methanol (30 mL x 3), and the filtrate was concentrated under reduced pressure to give methyl 2-amino-4-hydroxybenzoate 20d (1.3 g, yellow solid, yield: 81%).

[0355] MS (ESI): m / z 167.60 [M+H] + .

[0356] Step 5: Preparation of dimethyl 4,4'-(ethane-1,2-diylbis(oxy))bis(2- aminobenzoate) (20e)

[0357] Methyl 2-amino-4-hydroxybenzoate 20d (700 mg, 4.19 mmol) was dissolved in N,N-dimethylformamide (7 mL), and 1,2-dibromoethane (1.57 g, 8.38 mmol) and potassium carbonate (1.73 g, 12.6 mmol) were added. The reaction was stirred at 80 °C under nitrogen protection for 12 hours.

[0358] LCMS showed the reaction was completed, water (20 mL) was added to the reaction, and extraction was performed with ethyl acetate (20 mL x 3). The organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA:Et3N = 10:1:0.1% to 1:3:0.1%) to give dimethyl 4,4'-(ethane-1,2-diylbis(oxy))bis(2- aminobenzoate) 20e (380 mg, yellow solid, yield: 50%)

[0359] MS (ESI): m / z 361.20 [M+H] + .

[0360] 1 H NMR (300 MHz, DMSO-d6) d 7.63 (d, J = 9.0 Hz, 2H), 6.70 (br s, 4H), 6.31 (d, J = 2.4 Hz, 2H), 6.18 (dd, J = 8.7 Hz, 2.4 Hz, 2H), 4.26 (s, 4H), 3.74 (s, 6H).

[0361] Step 6: Preparation of 2,2'-((ethane-1,2-diylbis(oxy))bis(2-amino-4,1- phenylene))bis(propan-2-ol) (20)

[0362] To a solution of 4,4'-(ethane-1,2-diylbis(oxy))bis(2-aminobenzoic acid dimethyl ester) 20e (380 mg, 1.05 mmol) in tetrahydrofuran (4 mL) was cooled to -78 °C, 3.0 M methyl magnesium bromide solution (21 mL, 63 mmol) was added dropwise, the reaction was slowly warmed to room temperature and stirred for 12 hours.

[0363] LCMS indicated the reaction was complete, the reaction was slowly poured into ice water (20 mL) 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, concentrated under reduced pressure, the crude product was purified by silica gel column chromatography (PE:EA:Et3N = 2:1 :0.1 % to 1 :2:0.1 %) and then purified by prep (acetonitrile:0.1 % aqueous ammonium bicarbonate solution = 20% to 60%) to give 2,2'-((ethane-1,2-diylbis(oxy))bis(2-amino-4,1 - phenylene))bis(propan-2-ol) 20 (210 mg, white solid, yield: 56%).

[0364] MS (ESI): m / z 325.1 [M-2H20 + H] + .

[0365] 1 H NMR (400 MHz, DMSO-d6) d 6.89 (d, J = 8.8 Hz, 2H), 6.22 (d, J = 2.4 Hz, 2H), 6.08 (dd, J = 8.4 Hz, 2.8 Hz, 2H), 5.43 (br s, 4H), 5.09 (br s, 2H), 4.13 (s, 4H), 1.46 (s, 12H).

[0366] Example 21 : Preparation of compound 21

[0367] Step 1 : Preparation of intermediate 21 a

[0368] Ethyl triphenylphosphonium bromide (26.75 g, 72 mmol) was dissolved in tetrahydrofuran (200 mL) and purged with nitrogen three times. To the reaction was added potassium tert-butoxide (8.1 g, 72 mmol) portionwise at 0 °C. After 30 minutes, 5-formyl-2-nitrobenzoic acid methyl ester (10.0 g, 47.8 mmol) was added to the reaction and the reaction was slowly warmed to room temperature and stirred for 3 hours.

[0369] LCMS detection of reaction completion, the reaction liquid was added to saturated ammonium chloride (100 mL) dilution, and extracted with ethyl acetate (100 mL x 3), the organic phase was washed with saturated brine, anhydrous sodium sulfate drying, reduced pressure concentration. The crude product was purified by silica gel column chromatography (PE: EA = 15: 1) to obtain intermediate 21a (6.8 g, yellow oil, yield: 64.3%).

[0370] MS (ESI): m / z 222.2 [M+H] + .

[0371] Step 2: Preparation of intermediate 21b

[0372] To a solution of intermediate 21a (3.5 g, 15.8 mmol) in 1,2-dichloroethane (80 mL) was added bromine (5.81 g, 36.34 mmol) at room temperature. The reaction liquid was stirred at room temperature for 1 hour.

[0373] LCMS detection of reaction completion, the reaction liquid was diluted with dichloromethane (200 mL), saturated sodium bisulfite (30 mL x 3), saturated sodium chloride (50 mL x 3) washing, anhydrous sodium sulfate drying, reduced pressure concentration to obtain intermediate 21b (6.1 g, yellow oil, yield: 100%).

[0374] MS (ESI): m / z 401.0 [M+Na] + .

[0375] Step 3: Preparation of intermediate 21c

[0376] To a solution of intermediate 21b (6.1 g, 16 mmol) in tetrahydrofuran (30 mL) and methanol (30 mL) was added potassium hydroxide (1.8 g, 32 mmol). The reaction liquid was stirred at room temperature for 20 minutes.

[0377] LCMS detection of reaction completion, the reaction liquid was quenched with saturated ammonium chloride (80 mL), diluted with ethyl acetate (200 mL), washed with saturated sodium chloride (50 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE: EA, 30: 1-12: 1) to obtain intermediate 21c (1.7 g, colorless oil, yield: 35.4%).

[0378] MS (ESI): m / z 317.0 [M+H2O] + .

[0379] Step 4: Preparation of intermediate 21d

[0380] Intermediate 21c (1.7 g, 5.7 mmol), methyl 2-amino-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (1.9 g, 6.84 mmol), potassium carbonate (2.36 g, 17.1 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (420 mg, 0.57 mmol) were dissolved in water (2.0 mL) and 1,4-dioxane (20 mL) and purged with nitrogen for three times. The reaction was stirred at 100 °C for 3 h.

[0381] LCMS showed the reaction was completed, the reaction 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 21d (1.7 g, yellow oil, yield: 81.03%).

[0382] MS (ESI): m / z 371.1 [M+H] + .

[0383] Step 5: Preparation of intermediate 21e

[0384] Intermediate 21e was prepared from intermediate 21d (2.4 g, 6.5 mmol) following the procedure of Example 14, step 5, yield: 68%.

[0385] MS (ESI): m / z 341.2 [M+H] + .

[0386] Step 6: Preparation of compound 21

[0387] Compound 21 was prepared from intermediate 21e (1.5 g, 4.4 mmol) and 3.0 M methyl magnesium bromide solution (29.3 mL, 88 mmol) following the procedure of Example 14, step 6, yield: 51.3%.

[0388] MS (ESI): m / z 341.1 [M+H] + .

[0389] 1 H NMR (400 MHz, MeOD) d 6.98 (s, 1H), 6.90 (s, 1H), 6.86 (d, J = 2.0 Hz, 1H), 6.79 (d, J = 8.0 Hz, 1H), 6.71 (d, J = 8.0 Hz, 1H), 6.58 (d, J = 8.0 Hz, 1H), 5.89-5.84 (m, 1H), 1.71 (d, J = 6.8 Hz, 3H), 1.57 (s, 6H), 1.52 (s, 6H).

[0390] Example 22: Preparation of compound 22

[0391] Step 1: Preparation of intermediate 22a

[0392] Intermediate 22a was prepared from 5-bromo-2-nitrobenzoic acid methyl ester (2.5 g, 9.6 mmol) and 4,4,5,5-tetramethyl-2-(2-methylprop-1-en-1-yl)-1,3,2-dioxaborolane (2.62 g, 14.4 mmol) following the procedure of Example 14, step 4, yield: 92.92 %.

[0393] MS (ESI): m / z 236.2 [M+H] + .

[0394] Step 2: Preparation of intermediate 22b

[0395] Intermediate 22b was prepared from intermediate 22a (2.1 g, 8.927 mmol) and bromine (4.279 g, 26.78 mmol) following the procedure of Example 21, step 2, yield: 96.41 %.

[0396] MS (ESI): m / z 397.9 [M+H+2+2] + .

[0397] Step 3: Preparation of intermediate 22c

[0398] Intermediate 22c was prepared from intermediate 22b (3.4 g, 8.6066 mmol) following the procedure of Example 21, step 3, yield: 38.9 %.

[0399] MS (ESI): m / z 314.1 [M+H] + .

[0400] Step 4: Preparation of intermediate 22d

[0401] Intermediate 22d was prepared from intermediate 22c (900 mg, 2.865 mmol) and 2-amino-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid methyl ester (1.19 g, 4.3 mmol) following the procedure of Example 21, step 4, yield: 86.3 %.

[0402] MS (ESI): m / z 385.2 [M+H] + .

[0403] Step 5: Preparation of intermediate 22e

[0404] Intermediate 22e was prepared from intermediate 22d (550 mg, 1.43 mmol) following the procedure of Example 14, step 5, in yield: 74.8%.

[0405] MS (ESI): m / z 355.2 [M+H] + .

[0406] Step 6: Preparation of compound 22

[0407] Compound 22 was prepared from intermediate 22e (380 mg, 1.072 mmol) and 3.0 M methyl magnesium bromide solution (10 mL, 30.0 mmol) following the procedure of Example 14, step 6, in yield: 52.1%.

[0408] MS (ESI): m / z 355.1 [M+H] + .

[0409] 1 H NMR (400 MHz, DMSO-d6) δ 6.75 (d, J = 2.0 Hz, 2H), 6.58 (dd, J = 1.6 Hz, 8.0 Hz, 2H), 6.48 (d, J = 8.0 Hz, 2H), 5.33 (br s, 4H), 5.18 (s, 2H), 1.71 (s, 6H) 1.44 (s, 12H).

[0410] Example 23: Preparation of compound 23

[0411] Step 1: Preparation of intermediate 23a

[0412] Methyl 5-bromo-3-nitropyridinecarboxylate (6.5 g, 24.9 mmol), (E)-tert- butyldimethyl((3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)allyl)oxy)silane (9.66 g, 32.37 mmol), potassium carbonate (8.6 g, 62.25 mmol), l,l'-bis(diphenylphosphino)ferrocenedichloropalladium (II) (1.82 g, 2.49 mmol) were dissolved in 1,4-dioxane (60 mL) and water (6 mL), and purged with nitrogen three times. The reaction was stirred at 90 °C for 3 hours.

[0413] TLC showed the reaction was completed, the reaction was filtered, and 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 23a (7.0 g, yellow oil, yield: 75.9%).

[0414] MS (ESI): m / z 353.2 [M+H] + .

[0415] Step 2: Preparation of intermediate 23b

[0416] Intermediate 23a (7.0 g, 19.9 mmol) was dissolved in tetrahydrofuran (100 mL), 1.0 M aqueous hydrochloric acid (10 mL) was added, and the reaction was stirred at room temperature for 2 hours.

[0417] The reaction was detected by LCMS to be completed, and the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 8:1) to obtain intermediate 23b (5.0 g, yellow oil, yield: 94.97%).

[0418] MS (ESI): m / z 239.2 [M+H] + .

[0419] Step 3: Preparation of intermediate 23c

[0420] Intermediate 23b (5.0 g, 21.0 mmol) was dissolved in dichloromethane (50 mL). Phosphorus tribromide (2.84 g, 10.5 mmol) was added dropwise to the reaction solution at 0°C, and the reaction solution was slowly warmed to room temperature and stirred for 3 hours.

[0421] The reaction was detected by LCMS to be completed, and saturated aqueous sodium bicarbonate solution was added to the reaction solution, which was extracted with dichloromethane (50 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH = 50:1) to obtain intermediate 23c (3.0 g, yellow solid, yield: 45.24%).

[0422] MS (ESI): m / z 303.1 [M+H] + .

[0423] Step 4: Preparation of intermediate 23d

[0424] Intermediate 23c (3.0 g, 10 mmol) and 5-hydroxy-2-nitrobenzoic acid methyl ester (2.17 g, 11 mmol) were dissolved in dimethylformamide (30 mL). Potassium carbonate (3.46 g, 25 mmol) was added to the reaction solution, and the reaction solution was stirred at 80°C for 3 hours.

[0425] The reaction was detected by LCMS to be completed, and water was added to the reaction solution, which was extracted with ethyl acetate (50 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH = 30:1) to obtain intermediate 23d (2.3 g, yellow solid, yield: 52%).

[0426] MS (ESI): m / z 418.2 [M+H] + .

[0427] Step 5: Preparation of intermediate 23e

[0428] Intermediate 23e was prepared from intermediate 23d (2.0 g, 4.8 mmol) following the procedure of Example 14, Step 5, in a yield of 56.25%.

[0429] MS (ESI): m / z 358.2 [M+H] + .

[0430] Step 6: Preparation of compound 23

[0431] Compound 23 was prepared from intermediate 23e (1.0 g, 2.8 mmol) and 3.0 M methyl magnesium bromide solution (9 mL, 28 mmol) following the procedure of Example 14, Step 6, in a yield of 11.3%.

[0432] MS (ESI): m / z 358.1 [M+H] + .

[0433] 1 H NMR (400 MHz, MeOD) d 7.76 (s, 1H), 7.13 (s, 1H), 6.81 (s, 1H), 6.72-6.59 (m, 3H), 6.47-6.40 (m, 1H), 4.60 (d, J = 5.6 Hz, 1H), 1.59 (s, 6H) 1.58 (s, 6H).

[0434] Evaluation of in vitro activity

[0435] Example 24: In vitro aldehyde capturing ability experiment

[0436] Main principle of the experiment

[0437] The level of active aldehyde is found to be elevated in various inflammatory eye diseases, and if not promptly removed, these aldehydes can accelerate the symptoms of inflammation and worsen the eye disease. In this experiment, by simulating the in vivo environment, the relatively optimal compound is selected according to the complexing ability of the compound with the aldehyde in vivo.

[0438] Experimental materials and reagents

[0439] Sulfobutyl-β-cyclodextrin (Bide Pharmaceutica, BD243603-10g), linoleic acid (Adamas Reagent, 69208F), triolein (Sigma-Aldrich, T7140-10G), nonenal (Bide Pharmaceutica, BD19638-1g).

[0440] Experimental steps

[0441] 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.

[0442] 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 (20 μmol, 2.0 equivalent), and stir the reaction at 1000 rpm.

[0443] S3, add 1500 μl of methanol to the Eppendorf tube at 0 (no nonenal solution added to the reaction system, add 1510 μl of methanol), 10, 20, 40 and 60 minutes after stirring the reaction, respectively, high-speed vortex, then centrifuge quickly and transfer the solution to the high performance liquid chromatography sample analysis.

[0444] Experimental results

[0445] The liquid chromatography conditions are shown in Table 1 below:

[0446] Table 1: Liquid gradient conditions

[0447] Wherein, the sample injector temperature is 15℃; the column temperature is 40℃; the ultraviolet-visible light detection channels are shown in Table 2 below:

[0448] Table 2: Ultraviolet-visible light detection channels for Examples 1-20

[0449] Assuming that the light absorption coefficient of the complex product is consistent with the tested compound, the peak area of the tested compound in the T0 sample (sample without the addition of nonenal) is set as 100%, and the percentage of the complex product generation is calculated according to the ratio of the peak area of the main product peak at each time point to the starting peak area of the tested compound, to observe the ability of the compound to capture aldehyde. As can be seen from the ultraviolet-visible light detection channel table, nonenal has weak absorption at the maximum absorption wavelength of the complex product, and the influence on the content of the complex product can be ignored. The percentage content (%) of the complex product of the compound prepared in the examples of the present application is shown in Table 3 below:

[0450] Table 3: Percentage content (%) of complex product formed by the compound prepared in Examples 1-23

[0451] *: XRPR-7 is the preferred compound 3 in the patent CN113227051.

[0452] From the above table, it can be seen that the compounds prepared in the embodiments of the present application have very significant complexing aldehyde ability and speed, and can be used for treating or preventing diseases with RASP-mediated pathological characteristics, including but not limited to dry eye, Behcet's disease, Sjogren's syndrome, non-infectious uveitis and allergic conjunctivitis, etc.

[0453] Animal efficacy evaluation

[0454] Example 25: Efficacy test of allergic conjunctivitis animal model

[0455] 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. Inflammatory mediators and inflammatory cell-released proteases and other substances 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 symptoms 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-dropping C48 / 80, and investigates the improvement effect of the compounds of the present application on the rat conjunctivitis model.

[0456] Sample preparation: The positive drug uses commercially available olopatadine hydrochloride eye drops (0.1%, Alcon, VEY89A); 1% compound 23 eye drops are prepared by mixing citric acid sodium, tromethamine, glycerol, hydrochloric acid, sodium hydroxide, sodium chloride and water in a certain proportion.

[0457] Grouping: Female SD rats were used as research objects, and the SD rats were randomly grouped into normal control group, model control group, olopatadine hydrochloride group and compound 23 group. The normal control group was not modeled, and the other groups were modeled. The specific grouping information is shown in Table 4.

[0458] Table 4: Drug administration and dose design table of each group

[0459] Modeling, administration and evaluation: The rats were grouped according to random number method, the rats were anesthetized, the normal control group was given 10 μL / eye of physiological saline dropped into 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) dropped into the conjunctival sac, and the eye was gently closed for 10 s to prevent the solution from overflowing.

[0460] After 10 min of stimulation, 15 μL / eye of the test substance was taken and dropped into the conjunctival sac, and the eye was gently closed for 10 s to prevent the solution from overflowing, and the normal control group and the model control group were given the same volume of physiological saline. After 20 min of the third administration, 24 h after modeling, the eye examination was performed under a slit lamp, and the double eyes of each animal were individually clinically scored according to the scoring details in Table 5.

[0461] Table 5: Conjunctivitis scoring details of rats

[0462] According to the scoring details in Table 5, the score value is positively correlated with the severity of ocular inflammation. The experimental results are shown in Figure 1, and the compound 23 of the present application has a good therapeutic effect on C48 / 80-induced allergic conjunctivitis in 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 23 group.

[0463] 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 species inhibitor, characterized in that, comprising a compound having the general formula (I) or a derivative thereof, wherein, R 1 R is one or more substituents selected from the group consisting of hydrogen, hydroxy, cyano, nitro, halogen, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcarbonyl and C 1-6 alkylamino; X and Z are independently selected from C or N; Y is selected from -CH=CH-, -C≡C-, R1 is selected from H, CH3, or -CH2CH2CH2O-; R2, R3 are independently selected from H or CH3; 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 active aldehyde inhibitor according to claim 1, wherein R 1 at least one selected from the group consisting of hydrogen, hydroxy, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl and C 1-6 alkylamino.

3. The active aldehyde inhibitor of claim 1, wherein, The compounds of general formula (I) are selected from the group consisting of: 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.

4. A pharmaceutical composition, characterized by, The pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

5. The pharmaceutical composition of claim 4, 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.

6. The pharmaceutical composition of claim 4, wherein The pharmaceutical composition is used for preparing a medicine as an aldehyde capturing agent.

7. Use of an active aldehyde inhibitor according to any one of claims 1 to 3, characterized in that, The medicine is used for treating or preventing various diseases caused by active aldehyde substances.

8. Use according to claim 7, wherein the compound is ###0002### The disease refers to an eye disease.

9. Use according to claim 8, wherein the compound is ###0002### The eye disease comprises at least one of dry eye, allergic conjunctivitis, Behcet's disease, Sjogren's syndrome, and uveitis.

10. Use according to claim 9, wherein ​

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