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

An AI-designed inhibitor of active aldehydes and its application Technical Field

[0001] The present invention belongs to the field of drug synthesis and relates to an aldehyde capture agent, and specifically to an active aldehyde substance inhibitor designed based on AI and its application. Background Art

[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 an imbalance in the ocular surface microenvironment. It can be accompanied by ocular surface inflammatory reactions, tissue damage, and neurological abnormalities, resulting in a variety of ocular discomfort symptoms and / or visual dysfunction. The causes of dry eye are complex, and can be caused by environmental, lifestyle, immune, ocular surgery, medication, and age-related factors. Epidemiological surveys show that the global incidence of dry eye is 8%-34%, with Asia having the highest incidence of dry eye in the world, with the incidence in China reaching 21%-30%.

[0003] Reactive aidehyde species (RASPs), such as malondialdehyde (MDA) and 4-hydroxy-2-nonenal (HNE), are produced in vivo through diverse physiological processes. They covalently bind to amino and sulfhydryl groups on receptors and kinases, thereby amplifying upstream proinflammatory signaling pathways involving NF-κB, inflammasomes, scavenger receptor A, and other mediators. Elevated RASP levels are found in a variety of inflammatory eye diseases, including Behçet's disease, Sjögren's syndrome, noninfectious uveitis, allergic conjunctivitis, and dry eye. Studies have found elevated MDA levels in the tears of patients with dry eye, and the degree of MDA elevation is positively correlated with dry eye severity. Another study also demonstrated elevated MDA and HNE levels in tear fluid and conjunctival biopsies of dry eye patients compared with controls, and this level correlated with symptom severity. In addition to proinflammatory signaling pathways, RASPs also bind to phosphatidylethanolamine. Phosphatidylethanolamine is a key component of tear lipids and is crucial for water retention in ocular surface tissues. Therefore, RASP may be a potential therapeutic target for the treatment of dry eye.

[0004] RASP is closely associated with the pathogenesis of various diseases and has been shown to be a target for major diseases such as dry eye, allergic conjunctivitis, uveitis, and Sjögren's syndrome. Therefore, RASP inhibitors have broad application prospects as drugs. However, to date, no suitable RASP inhibitors have been marketed, and only a few are currently in clinical research. Therefore, the search for RASP inhibitors with higher RASP inhibitory activity and lower toxicity has significant theoretical significance and application value. Summary of the Invention

[0005] In order to overcome the defects in the prior art, the present invention screened and prepared an active aldehyde substance inhibitor through AI technology, which has an unexpected inhibitory effect on active aldehyde substances.

[0006] An object of the present invention is to provide an active aldehyde inhibitor comprising a compound represented by general formula (I) or a derivative thereof:

[0007] Among them, R 1 is one or more substituents selected from hydrogen, hydroxy, cyano, nitro, halogen, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, 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 Selected from hydrogen, hydroxy, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 At least one of alkylamino groups.

[0012] Optionally, the compound represented by general formula (I) is selected from:

[0013] At least one of .

[0014] Another object of the present invention is to provide a pharmaceutical composition comprising the compound represented by the above general formula (I) or its derivatives as an active ingredient.

[0015] Optionally, the pharmaceutical composition further comprises pharmaceutical excipients.

[0016] Optionally, the pharmaceutical composition is a detection reagent or kit, which comprises 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 represented by general formula (I), for identifying RASP ligands.

[0017] Another object of the present invention is to provide a use of the compound represented by the above general formula (I) and its derivatives for preparing a drug as an aldehyde trapping agent.

[0018] Alternatively, the drug can inhibit RASP and be used to treat or prevent various diseases caused by reactive aldehydes.

[0019] Optionally, the disease is an eye disease.

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

[0021] Beneficial effects of the present invention

[0022] The present invention prepares a compound represented by general formula (I), and unexpectedly finds that it has an unexpectedly good RASP inhibitory effect, and can be used to prepare a drug for treating or preventing diseases or conditions such as dry eye, Behcet's disease, Sjögren's syndrome, non-infectious uveitis and allergic conjunctivitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] FIG1 is a schematic diagram showing the results of the efficacy experiment on the rat allergic conjunctivitis animal model using Compound 23 in Example 25 of the present invention. DETAILED DESCRIPTION

[0024] The present invention is further illustrated below with reference to the accompanying drawings and examples, but the scope of protection of the present invention is not limited to these examples. Unless otherwise specified, the percentages described in the present invention are all weight percentages. The numerical ranges described in the specification, such as units of measurement, reaction conditions, physical states of compounds, or percentages, are all intended to provide unambiguous written references. When practicing this patent, those skilled in the art may use temperatures, concentrations, amounts, carbon number, etc. outside of these ranges or different from individual numerical values ​​and still obtain the expected results.

[0025] In the latest developments in AI technology, artificial intelligence can be used for the discovery, screening, and optimization of small molecule drugs. This is achieved by integrating AI computing tools, databases (such as the CAS compound library), and receptor-ligand complex interaction information into efficient molecular / atom training sets and AI digital workflows, and then combining them with BT technology (bioinformatics, in vitro target cell bioactivity assays, etc.) to invent new therapeutic drugs. As we all know, in the drug development process, traditional small molecule compounds (drugs) take a lot of time (usually 5-6 years or even longer) to progress from seed compounds to lead compounds and then to candidate compounds. However, drug development driven by artificial intelligence (AIDrug Discovery & Design, AIDD) can perform data cross-comparison, molecular docking, molecular dynamics simulations, etc. in a shorter period of time (usually only 3-5 months), thereby accelerating the screening or de novo design of new compounds. Its core value is reflected in groundbreaking innovation and efficiency improvements.

[0026] Based on relatively clear mechanisms (such as the Siedinger equation and Gibbs free energy change), AI can search multiple, more extensive and comprehensive databases of different compounds and proteomics, rapidly completing searches and cross-comparisons, saving a significant amount of investment originally spent on wet experiments. Wet experiment data can then be fed back to the AI ​​for iterative optimization of compound structures.

[0027] Based on NLP (Natural Language Processing), in-depth analysis of disease / drug similarity networks, and molecular generation model training, the present invention independently builds an AI artificial intelligence server and workstation, which can simultaneously and independently perform deep learning and multi-threaded collaborative simulation calculations on multiple drugs, and can also be formed into an AI computing cluster to process candidate compound big data, deeply explore new drug targets, save innovative drug development costs, and shorten R&D time; thus forming a new R&D path for new drug R&D - "dry-wet combination, seamless connection", which is expected to become a scientific paradigm for innovative drug R&D in the future, and bring new ideological breakthroughs and experimental innovations to the previous drug R&D process that was extremely dependent on wet experiments, which was time-consuming, labor-intensive, and cumbersome.

[0028] The present invention is based on the above-mentioned independently built AI technology platform. By calculating and predicting the alkalinity of aromatic amines, the transition state activation energy ΔG of imine reactions, and the chemical reaction enthalpy change ΔH, and then combining iterative optimization with wet experiments, a series of active aldehyde substance inhibitors in the present invention are finally obtained.

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

[0030] The basic mechanism of action of RSAP inhibitors is that the amino group in the structure reacts with reactive aldehydes, thereby reducing the concentration of reactive aldehydes in target tissues. The amino group reacts with aldehydes to form imines. The reaction mechanism is as follows:

[0031] The aldehyde carbonyl group contains a carbon-oxygen double bond. Since the mobile π electrons are strongly pulled toward 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 in the carbonyl group nucleophilically, forming an unstable amine imine, which removes a molecule of water to form a carbon-nitrogen double bond (imine). The stronger the alkalinity of the amine, the easier it is to undergo the nucleophilic reaction. The alkalinity 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 invention are all aromatic amines. Specific substituents on the aromatic ring have an effect on the alkalinity of the amine. Electron-pushing groups push electrons toward nitrogen, enhancing the alkalinity of the aromatic amine, allowing the lone pair of electrons to react more effectively with the electron-deficient carbon; while electron-withdrawing groups help pull electrons away from nitrogen, weakening the alkalinity of the aromatic amine, making it difficult for the lone pair of electrons to react with the electron-deficient carbon. Common electron-pushing groups include amino, hydroxyl, alkoxy, alkyl, phenyl, alkylamino, etc., and common electron-withdrawing groups include halogen, nitro, cyano, alkylcarbonyl, haloalkyl, etc. Specifically speaking, in the definition of R1, the substituents on the aromatic amines in Examples 5, 6, and 7 of the present invention are halogens, which are electron-withdrawing groups. These three examples have aldehyde-capturing capabilities. The cyano, nitro, C1-6 haloalkyl, and C1-6 alkylcarbonyl groups in the definition of R1 are all electron-withdrawing groups. It is reasonable to infer theoretically that compounds with such substituents have similar aldehyde-capturing capabilities as those in Examples 5, 6, and 7; the hydroxyl, C1-6 alkyl, C1-6 alkoxy, and C1-6 alkylamino groups in the definition of R1 are all electron-pushing groups. It is reasonable to infer theoretically that compounds with such substituents have better aldehyde-capturing capabilities than those in Examples 5, 6, and 7.

[0032] Therefore, R 1 It can be one or more substituents selected from hydrogen, hydroxy, cyano, nitro, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Alkylcarbonyl and C 1-6 Alkylamino.

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

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

[0035] The definition of L in the compound provided by Chinese patent CN113227051 is single bond, -O-, -S-, -NR2, and -(CR3R4)n-, which are all electron-donating groups that increase the alkalinity of aromatic amines and are beneficial to imine reactions. The -CH=CH-, -C≡C-, All of them are electron-withdrawing groups, which reduce the alkalinity of aromatic amines and are not conducive to imine reactions. Theoretically, when Y takes the above electron-withdrawing groups, the aldehyde-capturing ability of the compounds of the present invention should be weaker than that of the compounds in patent CN113227051. However, the inventors unexpectedly found that the aldehyde-capturing ability of the compounds of the present invention is stronger than that of the preferred compounds in patent CN113227051, which has an unexpected technical effect.

[0036] The "halogen" mentioned in the present invention refers to fluorine, chlorine, bromine and iodine.

[0037] The "C 1-6 “Alkyl” refers to straight-chain, 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, tert-butyl, neopentyl, isopentyl, isohexyl, and 2,2-dimethylpropyl; examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

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

[0039] The "alkoxy group" mentioned in the present invention refers to a group formed by the above-mentioned alkyl group and an oxygen atom, wherein the oxygen atom has the ability to form a free bond, such as methoxy, ethoxy, propoxy, butoxy, pentyloxy, isopropoxy, tert-butoxy, cyclopropyloxy, cyclohexyloxy, etc.

[0040] The "pharmaceutically acceptable salts" or "pharmaceutically usable salts" of the present invention refer to pharmaceutically acceptable acid and base addition salts and solvates. Such pharmaceutically acceptable salts include salts formed with acids such as hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, sulfinic acid, formic acid, p-toluenesulfonic acid, methanesulfonic acid, nitric acid, benzoic acid, citric acid, tartaric acid, maleic acid, hydroiodic acid, chain carboxylic acids such as acetic acid, HOOC-(CH2) n -COOH (n = 0-4) and the like; also includes salts formed with bases, the cations of these salts include sodium, potassium, calcium, ammonium ions and the like.

[0041] In the present invention, "substituted" means that the organic group defined herein (which contains one or more bonds to hydrogen atoms) is replaced by one or more bonds to non-hydrogen atoms or atomic groups, and the non-hydrogen atoms or atomic groups are substituents.

[0042] In the present invention, "derivative" refers to a compound formed by replacing an atom or an atomic group in the compound represented by general formula (I) with another atom or an atomic group, including at least one of a fluorescent label, a spin label, a heavy metal label, an isotope label and a pharmaceutically acceptable salt.

[0043] In another aspect, the present invention also relates to fluorescently labeled, spin-labeled, heavy metal-labeled, or isotopically labeled derivatives of the compounds described herein, which can be used not only for imaging but also for in vivo and in vitro detection, localization, and quantification of RASP in tissue samples (including humans) by inhibiting binding of the labeled compound and identifying RASP ligands. Accordingly, the present invention further provides RASP detection reagents or kits containing such labeled compounds.

[0044] The present invention further provides isotopically labeled compounds of the compounds of the present invention. The "isotopically labeled compounds" or "isotopically labeled" compounds of the present invention refer to compounds described herein in which 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 typically found in nature (i.e., naturally occurring). Applicable radionuclides may 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 type of radioisotope contained in the isotopically labeled compound will depend on the specific application of the isotopically labeled compound. For example, for in vitro labeling and competition assays of IDO enzymes, compounds containing 3H, 14C, 82Br, 125I, 131I, 35S are generally most useful. For isotope 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 radioactive isotopes are also applicable to the compounds of the present invention.

[0046] When administered as a drug, the compound can be administered in the form of a pharmaceutical composition. Therefore, in another aspect, the present application provides a pharmaceutical composition containing the compound or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.

[0047] The term "composition" is used to refer to a product containing a compound disclosed in this application or a pharmaceutically acceptable salt thereof as the specific active ingredient, as well as any other product that is in combination with the active ingredient directly or indirectly.

[0048] Usually, the pharmaceutical composition contains at least one pharmaceutically acceptable carrier or excipient. The term "pharmaceutically acceptable" means that the carrier or excipient is compatible with the other ingredients in the formula and is harmless to the subject. The carrier described here refers to a substance used to improve the selectivity, effectiveness and / or safety of the drug during delivery. The carrier is mainly used to control drug release and can also be used to improve the pharmacokinetic properties of the drug, especially bioavailability. The excipient refers to other substances in the pharmaceutical preparation other than the active ingredient, which are mainly used for long-term stability, filling solid preparations (therefore, it is often used to specifically refer to "fillers") or enhancing product efficacy (for example, promoting absorption, reducing viscosity or increasing solubility, etc.).

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

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

[0051] HATU: 2-(7-benzotriazole oxide)-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 combined with mass spectrometry detection

[0062] TLC detection: thin layer chromatography detection.

[0063] Example 1: Preparation of Compound (E)-2,2′-(ethylene-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-bromopicolinate (2.5 g, 10.87 mmol) was dissolved in tetrahydrofuran (30 mL) and cooled to -78°C. A 1.6 M solution of methylmagnesium bromide in tetrahydrofuran (27.2 mL, 43.48 mmol) was then slowly added dropwise. The reaction mixture was warmed to 0°C and stirred under nitrogen for 3 hours. LCMS and TLC confirmed the reaction was complete. At 0°C, saturated aqueous ammonium chloride (50 mL) was slowly added dropwise. The mixture was 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. The mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA = 3:1, volume ratio) to afford 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)propanol (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)ferrocenepalladium(II) dichloride (190 mg, 0.26 mmol), and cesium carbonate (2.1 g, 6.5 mmol) were dissolved in a mixture of 1,4-dioxane (20 mL) and water (4 mL). The reaction mixture was stirred at 90°C under nitrogen for 2 hours.

[0069] The reaction was complete after LCMS analysis. Water (30 mL) was added to the reaction solution, and the mixture was 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, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 5:1) to afford 2-(3-amino-5-vinylpyridin-2-yl)propanol 1b (450 mg, yield: 95%).

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

[0071] Step 3: Preparation of compound (E)-2,2′-(ethylene-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 mixture was stirred at 130°C under nitrogen for 16 hours.

[0073] The reaction was complete after LCMS detection. Water (30 mL) was added to the reaction solution and extracted with ethyl acetate (30 mL × 3). The organic phase was washed with saturated sodium chloride solution (30 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 = 10% to 60%) to give compound (E)-2,2′-(ethylene-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 (400MHz, DMSO-d6) δ7.87 (d, J=2.0Hz, 2H), 7.17 (d, J=2.0Hz, 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] Dissolve methyl 3-amino-6-bromopicolinate (500 mg, 2.17 mmol) in tetrahydrofuran (10 mL), cool to -78°C, then slowly add 1.6 M methyllithium tetrahydrofuran solution (5.4 mL, 8.68 mmol) dropwise. The reaction mixture is heated to 0°C and stirred under nitrogen for 3 hours.

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

[0080] 1 H NMR (300MHz, CDCl3) δ7.15 (d, J=8.4Hz, 1H), 6.83 (d, J=8.1Hz, 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 mixture was stirred at 130°C under nitrogen for 16 hours.

[0083] The reaction was complete after LCMS detection, and water (30 mL) was added to the reaction solution and extracted with ethyl acetate (30 mL×3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by preparative (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)pyridin-2-yl)propanol 2 (49 mg, yield: 9%).

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

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

[0086] Example 3: Preparation of Compound (E)-2,2′-(ethylene-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)ferrocenepalladium(II) dichloride (124 mg, 0.17 mmol), and cesium carbonate (1.4 g, 4.35 mmol) were dissolved in a mixture of 1,4-dioxane (20 mL) and water (4 mL). The reaction mixture was stirred at 90°C under nitrogen for 2 hours.

[0089] The reaction was complete after LCMS analysis. 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, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 5:1) to afford 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′-(ethylene-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 mixture was stirred at 130°C under nitrogen for 16 hours.

[0093] The reaction was complete after LCMS analysis. Water (10 mL) was added to the reaction solution and extracted with ethyl acetate (20 mL × 3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by preparative chromatography (acetonitrile: 0.1% aqueous ammonium bicarbonate solution = 10% to 60%) to give (E)-2,2′-(ethylene-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 (400MHz, DMSO-d6) δ7.12 (d, J=9.2Hz, 4H), 6.91 (d, J=8.0Hz, 2H), 5.64 (s, 4H), 5.49 (s, 2H), 1.53 (s, 12H).

[0096] 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), and potassium ethylene trifluoroborate (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. The mixture was heated to 90° C. and stirred for 5 hours under nitrogen protection.

[0099] The reaction was complete after LCMS analysis. Water (30 mL) was added to the reaction solution, and the mixture was 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, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1) to afford methyl 2-amino-4-vinylbenzoate 4a (600 mg, yellow oil, yield: 78%).

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

[0101] 1H NMR (400MHz, CDCl3) δ7.81 (d, J=8.4Hz, 1H), 6.74 (dd, J=8.4Hz, 1.6Hz, 1H), 6.66 (d, J=1.6Hz, 1H) , 6.65-6.58 (m, 1H), 5.79 (dd, J=17.2Hz, 0.4Hz, 1H), 5.33 (dd, J=10.8Hz, 0.4Hz, 1H), 3.87 (s, 3H).

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

[0103] Methyl 2-amino-4-vinylbenzoate 4a (200 mg, 1.13 mmol), methyl 3-amino-5-bromopicolinate (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 mixture was stirred at 130°C under nitrogen for 16 hours.

[0104] The reaction was complete after LCMS detection, and water (20 mL) was added to the reaction solution and extracted with ethyl acetate (20 mL × 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 reverse phase column (acetonitrile:water = 5% to 95%) to give ((E)-3-amino-5-(3-amino-4-(methoxycarbonyl)phenyl)picolinic acid methyl ester 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)phenylvinyl)picolinate 4b (130 mg, 0.397 mmol) was dissolved in tetrahydrofuran (2 mL) and cooled to -78°C. 3M methylmagnesium bromide solution in tetrahydrofuran (1.3 mL, 3.9 mmol) was then slowly added dropwise. The reaction mixture was warmed to 0°C and stirred under nitrogen for 2 hours.

[0108] The reaction was complete after LCMS detection, and water (30 mL) was added to the reaction solution and extracted with ethyl acetate (30 mL×3). The organic phase was washed with saturated sodium chloride solution (30 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 = 40% to 75%) to give ((E)-2-(2-amino-4-(2-amino-6-(2-hydroxypropyl-2-yl)pyridin-3-vinyl)-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 (400MHz, CD3OD) δ7.86 (d, J=2.0Hz, 1H), 7.24 (d, J=1.6Hz, 1H), 7.13 (d, J=8.0Hz, 1H), 7..04 (d, J=16.4Hz , 1H), 6.96 (d, J=16.4Hz, 1H), 6.90 (d, J=2.0Hz, 1H), 6.84 (dd, J=8.0Hz, 2.0Hz, 1H), 1.62 (s, 6H), 1.61 (s, 6H).

[0111] 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), and potassium ethylene trifluoroborate (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. The mixture was heated to 90° C. and stirred for 2 hours under nitrogen protection.

[0114] The reaction was complete after LCMS analysis. The reaction solution was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography (PE:EA=20:1) to give methyl 2-amino-5-fluoro-4-vinylbenzoate 5a (670 mg, yellow oil, yield: 85.2%).

[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 mixture was stirred at 130°C under nitrogen for 16 hours.

[0118] The reaction was completed by LCMS, and the reaction solution 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] Methyl (E)-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), cooled to 0°C, and then a 1.0 M solution of methylmagnesium bromide in tetrahydrofuran (6.1 mL, 6.09 mmol) was slowly added dropwise. The reaction mixture was stirred under nitrogen for 2 hours.

[0122] The reaction was complete after LCMS detection, and water (30 mL) was added to the reaction solution and extracted with ethyl acetate (30 mL×3). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. 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 (400MHz, CD3OD) δ7.88 (d, J=8.8Hz, 1H), 7.59-7.70 (m, 1H), 7.58 (d, J=8 .8Hz, 1H), 7.34-7.41(m, 2H), 7.08-7.11(m, 1H), 1.72(s, 6H), 1.65(s, 6H).

[0125] 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), and potassium ethylene trifluoroborate (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. The mixture was heated to 90° C. and stirred for 5 hours under nitrogen protection.

[0128] The reaction was complete after LCMS analysis. Water (30 mL) was added to the reaction solution, and the mixture was 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, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1) to afford 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 (400MHz, CDCl3) δ7.52 (d, J=11.6Hz, 1H), 6.80-6.73 (m, 2H), 5.87 (dd, J=18.0Hz, 0.8Hz, 1H), 5.45 (d, J=11.2Hz, 1H), 3.87 (s, 3H).

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

[0132] Methyl 2-amino-5-fluoro-4-vinylbenzoate 6a (200 mg, 1.02 mmol), methyl 3-amino-5-bromopicolinate (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 mixture was stirred at 130°C under nitrogen for 16 hours.

[0133] The reaction was complete after LCMS detection, and water (20 mL) was added to the reaction solution and extracted with ethyl acetate (20 mL × 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 reverse phase column (acetonitrile:water = 5% to 95%) to give methyl 3-amino-5-(5-amino-2-fluoro-4-(methoxycarbonyl)phenyl)picolinate 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)phenyl)picolinate 6b (130 mg, 0.376 mmol) was dissolved in tetrahydrofuran (2 mL) and cooled to -78°C. A 3M solution of methylmagnesium bromide in tetrahydrofuran (1.3 mL, 3.9 mmol) was then slowly added dropwise. The reaction mixture was warmed to 0°C and stirred under nitrogen for 2 hours.

[0137] The reaction was complete after LCMS analysis. Water (30 mL) was added to the reaction solution and the mixture was 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, and concentrated under reduced pressure. The crude product was purified by preparative chromatography (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 approximately 25% of the cis isomer).

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

[0139] 1 H NMR (400MHz, CD3OD) δ7.86 (d, J=2.0Hz, 1H), 7.25 (d, J=1.6Hz, 1H), 7.18 (d, J=16.4Hz, 1H), 7. 04 (d, J=16.8Hz, 1H), 6.98 (d, J=6.8Hz, 1H), 6.90 (d, J=12.8Hz, 1H), 1.61 (s, 6H), 1.60 (s, 6H).

[0140] Example 7: Synthesis of Compound ((E)-2-(2-amino-4-(2-amino-6-(2-hydroxypropyl-2-yl)pyridin-3-vinyl)-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 mixture 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 mixture was stirred at room temperature for 1.5 hours.

[0143] The reaction was complete after LCMS analysis. The reaction mixture was added with water (30 mL) and extracted with ethyl acetate (30 mL x 3). The mixture was washed with saturated sodium chloride solution (30 mL) and dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA = 10:1 to 8:1) to obtain methyl 2-amino-4-bromo-5-chlorobenzoate 7a (900 mg, white solid, yield: 57%).

[0144] 1 H NMR (300MHz, CDCl3) δ7.94 (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), cooled to -78°C, and then 3M methylmagnesium bromide solution in tetrahydrofuran (4.4 mL, 13.3 mmol) was slowly added dropwise. The reaction mixture was heated to 0°C and stirred under nitrogen for 3 hours.

[0147] TLC indicated the reaction was complete. Saturated aqueous ammonium chloride (30 mL) was slowly added dropwise to the reaction mixture at 0°C. The mixture was extracted with ethyl acetate (30 mL x 3), washed with saturated sodium chloride (30 mL), and dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure, and 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 (300MHz, CDCl3) δ7.09 (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-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 mixture was stirred at 130°C under nitrogen for 16 hours.

[0151] The reaction was complete after LCMS detection, and water (30 mL) was added to the reaction solution and extracted with ethyl acetate (30 mL×3). The organic phase was washed with saturated sodium chloride solution (30 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 = 10% to 60%) to give ((E)-2-(2-amino-4-(2-amino-6-(2-hydroxypropyl-2-yl)pyridin-3-vinyl)-5-chlorophenyl)propan-2-ol (57 mg, light yellow solid, yield: 12%).

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

[0153] 1H NMR (400MHz, DMSO-d6) δ7.82 (d, J=1.6Hz, 1H), 7.24 (d, J=16.4Hz, 1H), 7.21 (d, J=1.6Hz, 1H), 7.05 (d, J= 20.4Hz, 2H), 6.93 (d, J=16.4Hz, 1H), 5.63 (s, 2H), 5.54 (s, 2H), 5.46 (s, 1H), 5.34 (s, 1H), 1.49 (s, 12H).

[0154] Example 8: 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 (8)

[0155] Step 1: Preparation of (E)-4,4′-(ethylene-1,2-diyl)dimethylbis(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 the atmosphere was replaced with nitrogen three times. The reaction mixture was stirred at 130°C for 16 hours.

[0157] The reaction was complete after LCMS analysis and the product was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=10:1 to 1:2) to afford (E)-4,4′-(ethylene-1,2-diyl)dimethylbis(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′-(ethylene-1,2-diylbis(2-amino-4,1-phenylene))bis(propan-2-ol) (8)

[0160] (E)-4,4′-(ethylene-1,2-diyl)dimethylbis(2-aminobenzoate) 8a (500 mg, 1.53 mmol) was dissolved in tetrahydrofuran (20 mL), cooled to -78°C, and 3.0 M methylmagnesium bromide solution (15.3 mL, 46.0 mmol) was added dropwise. The temperature was slowly raised to room temperature and stirred for 12 hours.

[0161] The reaction was complete after LCMS detection. Saturated ammonium chloride (20 mL) was added to the reaction solution and extracted with ethyl acetate (40 mL × 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 (E)-2,2′-(ethylene-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 (400MHz, DMSO-d6) δ7.99 (d, J=8.0Hz, 2H), 6.86 (s, 2H), 6.79 (d, J=5.6Hz, 2H), 6.68 (dd, J1=1.6Hz, J2=8.0Hz, 2H), 5.42 (br s, 4H), 5.19 (s, 2H), 1.49 (s, 12H).

[0164] Example 9: Preparation of Compound (E)-2-(2-amino-4-(4-amino-3-(2-hydroxypropan-2-yl)phenyl)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 vinyl trifluoroborate (1.3 g, 9.78 mmol), 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (358 mg, 0.489 mmol), and triethylamine (3.6 mL, 26.1 mmol) were dissolved in ethanol (20 mL), and the atmosphere was purged with nitrogen three times. The reaction mixture was stirred at 90°C for 5 hours.

[0167] The reaction was complete after LCMS analysis and the residue was 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: (E) Preparation of methyl 2-amino-4-(4-amino-3-(methoxycarbonyl)phenylvinyl)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). The atmosphere was replaced with nitrogen three times. The reaction mixture was stirred at 120°C for 16 hours.

[0171] The reaction was complete after LCMS analysis and the product was 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)phenylvinyl)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)phenyl)phenyl)propan-2-ol (9)

[0174] Methyl (E) 2-amino-4-(4-amino-3-(methoxycarbonyl)phenylvinyl)benzoate 9b (300 mg, 0.919 mmol) was dissolved in tetrahydrofuran (20 mL), cooled to -78°C, and 3.0 M methylmagnesium bromide solution (7.7 mL, 23.0 mmol) was added dropwise. The mixture was slowly warmed to room temperature and stirred for 12 hours.

[0175] The reaction was complete after LCMS detection. Saturated ammonium chloride (20 mL) was added to the reaction solution and extracted with ethyl acetate (40 mL × 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 (E)-2-(2-amino-4-(4-amino-3-(2-hydroxyprop-2-yl)phenylvinyl)phenyl)propan-2-ol 9 (161 mg, yield: 53.7%).

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

[0177] 1H NMR (400MHz, DMSO-d6) δ7.19 (m, 2H), 6.96 (d, J = 8.0Hz, 1H), 6.88 (d, J = 16.0Hz, 1H ), 6.72(m, 2H), 6.64(dd, J1=1.6Hz, J2=8.0Hz, 1H), 6.60(d, J=8.4Hz, 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] 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)picolinate (10a)

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

[0181] The reaction was complete after LCMS analysis, and the product was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=10:1 to 4:1) to give methyl 3-amino-5-((trimethylsilyl)ethynyl)picolinate 10a (1.5 g, brown solid, yield: 82%).

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

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

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

[0185] The reaction was complete after LCMS analysis. Water (20 mL) was added to the reaction mixture, and the mixture was 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 afford methyl 3-amino-5-ethynylpicolinate 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)picolinate (10c)

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

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

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

[0191] Step 4: 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), cooled to -70°C, and 3.0 M methylmagnesium bromide solution (6.1 mL, 18.4 mmol) was added dropwise. The mixture was slowly warmed to room temperature and stirred for 12 hours.

[0193] The reaction was complete after LCMS detection. Saturated ammonium chloride (20 mL) was added to the reaction solution and extracted with ethyl acetate (20 mL × 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 2-(3-amino-5-((5-amino-6-(2-hydroxyprop-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 (400MHz, DMSO-d6) δ7.81 (d, J=2.0Hz, 1H), 7.21 (d, J=8.0Hz, 1H), 7.06 (d, J=2.0Hz, 1H), 6.94 (d, J=8.4Hz, 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] 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] Under nitrogen, ethyltrimethylsilane (1.54 g, 15.65 mmol) and triethylamine (3.6 mL, 26.1 mmol) were added sequentially to a solution of methyl 2-amino-4-bromobenzoate (1.2 g, 5.22 mmol), cuprous iodide (99.4 mg, 0.522 mmol), and bistriphenylphosphine palladium dichloride (366 mg, 0.522 mmol) in 1,4-dioxane (30 mL). The reaction mixture was stirred at 90° C. for 5 hours.

[0199] The reaction was complete after LCMS analysis and the residue was 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) and the reaction mixture was stirred at room temperature for 0.5 hours.

[0203] The reaction was complete after LCMS analysis. 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, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1) to afford methyl 2-amino-4-ethynylbenzoate 11b (750 mg, off-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), bistriphenylphosphine palladium dichloride (152 mg, 0.216 mmol), and triethylamine (1.5 mL, 10.8 mmol) were dissolved in 1,4-dioxane (20 mL), and the atmosphere was replaced with nitrogen three times. The reaction mixture was stirred at 95°C for 16 hours.

[0207] The reaction was complete after LCMS analysis, and the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 4:1 to 1:1) to afford 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), cooled to -78°C, and 3.0 M methylmagnesium bromide solution (10.2 mL, 30.7 mmol) was added dropwise. The mixture was slowly warmed to room temperature and stirred for 12 hours.

[0211] The reaction was complete after LCMS detection. Saturated ammonium chloride (20 mL) was added to the reaction solution and extracted with ethyl acetate (40 mL × 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 2-(2-amino-4-((5-amino-6-(2-hydroxyprop-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] 1 H NMR (400MHz, DMSO-d6) δ7.80 (d, J=2.0Hz, 1H), 7.05 (d, J=2.0Hz, 1H), 7.06 (d, J =8.0Hz, 1H), 6.78 (d, J = 2.0Hz, 1H), 6.65 (dd, J1 = 1.6Hz, J2 = 8.0Hz, 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] Example 12: Preparation of Compound 2-(2-amino-4-((4-amino-3-(2-hydroxyprop-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] 1-(2-Amino-5-iodophenyl)ethanone (520 mg, 1.92 mmol), methyl 2-amino-4-ethynylbenzoate 11b (436 mg, 2.49 mmol), cuprous iodide (36.6 mg, 0.192 mmol), bistriphenylphosphine palladium dichloride (135 mg, 0.192 mmol), and triethylamine (1.38 mL, 9.96 mmol) were dissolved in 1,4-dioxane (20 mL), and the atmosphere was replaced with nitrogen three times. The reaction mixture was stirred at 50°C for 16 hours.

[0217] The reaction was complete after LCMS analysis, and the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 5:1 to 3:1) to afford 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), cooled to -78°C, and 3.0 M methylmagnesium bromide solution (8.6 mL, 25.9 mmol) was added dropwise. The mixture was slowly warmed to room temperature and stirred for 12 hours.

[0221] The reaction was complete after LCMS detection. Saturated ammonium chloride (20 mL) was added to the reaction solution and extracted with ethyl acetate (40 mL × 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-((4-amino-3-(2-hydroxypropyl-2-yl)phenyl)ethynyl)phenyl 12 (230 mg, white solid, yield: 54.6%).

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

[0223] 1H NMR (400MHz, DMSO-d6) δ7.11 (d, J=1.6Hz, 1H), 7.07 (dd, J1=2.0Hz, J2=8.4Hz, 1H), 6.98 (d, J=8.0Hz, 1H), 6.71 (d, J=1.6Hz, 1H), 6.59 (m, 2H), 5.81 (br s, 2H), 5.50 (brs, 2H), 5.30 (s, 1H), 5.25 (s, 1H), 1.50 (s, 6H), 1.48 (s, 6H).

[0224] Example 13: Preparation of Compound 2,2′-(Ethylene-1,2-diylbis(2-amino-4,1-phenylene))bis(propan-2-ol) (13)

[0225] Step 1: Preparation of 4,4′-(ethynyl-1,2-diyl)bis(dimethyl 2-aminobenzoate) (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), bistriphenylphosphine palladium dichloride (152 mg, 0.217 mmol), and triethylamine (1.51 mL, 10.86 mmol) were dissolved in 1,4-dioxane (20 mL), and the atmosphere was replaced with nitrogen three times. The reaction mixture was stirred at 95°C for 16 hours.

[0227] The reaction was complete after LCMS analysis, and 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 afford dimethyl 4,4′-(ethynyl-1,2-diyl)bis(2-aminobenzoate) 13a (480 mg, yellow solid, yield: 68.2%).

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

[0229] Step 2: Preparation of 2,2′-(ethynyl-1,2-diylbis(2-amino-4,1-phenylene))bis(propan-2-ol) (13)

[0230] Dimethyl 4,4′-(ethynyl-1,2-diyl)bis(2-aminobenzoate) (480 mg, 1.48 mmol) was dissolved in tetrahydrofuran (20 mL), cooled to -78°C, and 3.0 M methylmagnesium bromide solution (9.9 mL, 29.6 mmol) was added dropwise. The mixture was slowly heated to room temperature and stirred for 12 hours.

[0231] The reaction was complete after LCMS detection. Saturated ammonium chloride (20 mL) was added to the reaction solution and extracted with ethyl acetate (40 mL × 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'-(acetylene-1,2-diylbis(2-amino-4,1-phenylene))bis(propan-2-ol) 13 (190 mg, white solid, yield: 39.6%).

[0232] MS (ESI): m / z 307.1 [M-H2O+H] + .

[0233] 1 H NMR (400MHz, DMSO-d6) δ7.01 (d, J=8.0Hz, 2H), 6.75 (d, J=2.0Hz, 2H), 6.62 (dd, J1=2.0Hz, J2=8.0Hz, 2H), 5.55 (br s, 4H), 5.28 (s, 2H), 1.49 (s, 12H).

[0234] Example 14: Preparation of Compound 2,2′-(Ethylene-1,1-dibis(2-amino-4,1-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 vinyl trifluoroborate (1.75 g, 13.08 mmol), 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (422 mg, 0.577 mmol), and triethylamine (5.3 mL, 38.45 mmol) were dissolved in ethanol (25 mL), and the atmosphere was replaced with nitrogen three times. The reaction mixture was stirred at 95°C for 4 hours.

[0237] The reaction was complete after LCMS analysis and the residue was 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, and the reaction mixture was stirred at room temperature for 1 hour.

[0241] The reaction was completed by LCMS. The reaction solution was diluted with dichloromethane (150 mL), washed with saturated sodium bisulfite (40 mL × 3) and saturated sodium chloride (40 mL × 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 (400MHz, DMSO-d6) δ8.16 (d, J=2.0Hz, 1H), 7.94 (dd, J1=2.4Hz, J2=10.8Hz, 1H), 7.87 (d, J=10.8Hz, 1H), 5.53 (dd, J1=3.2Hz, J2=10.0Hz, 1H), 4.33-4.20 (m, 2H), 3.95 (s, 3H).

[0244] Step 3: Preparation of methyl 4-(1-bromovinyl)-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) and the reaction was stirred at room temperature for 30 minutes.

[0246] The reaction was complete by LCMS. The reaction solution was diluted with ethyl acetate (200 mL), washed with saturated sodium chloride (40 mL × 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 afford methyl 4-(1-bromovinyl)-2-nitrobenzoate 14c (1.9 g, yellow oil, yield: 93.6%).

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

[0248] 1H NMR (400MHz, DMSO-d6) δ8.28 (d, J=2.0Hz, 1H), 8.09 (dd, J1=2.0Hz, J2=8.0Hz, 1H) , 7.95 (d, J=8.4Hz, 1H), 6.76 (d, J=3.2Hz, 1H), 6.16 (d, J=3.2Hz, 1H), 3.87 (s, 3H).

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

[0250] Methyl 4-(1-bromovinyl)-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)ferrocenepalladium(II) dichloride were dissolved in water (1.0 mL) and 1,4-dioxane (15 mL). The atmosphere was purged with nitrogen three times. The reaction mixture was stirred at 95°C for 4 hours.

[0251] The reaction was complete by LCMS. The reaction solution was cooled to room temperature, diluted with ethyl acetate (150 mL), washed with saturated sodium chloride (40 mL × 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 afford 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-(1-(4-(methoxycarbonyl)-3-aminophenyl)vinyl)benzoate (14e)

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

[0255] The reaction was complete by LCMS. The reaction solution was cooled to room temperature, diluted with ethyl acetate (150 mL), washed with saturated sodium chloride (30 mL × 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 afford methyl 2-amino-4-(1-(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′-(ethylene-1,1-dibis(2-amino-4,1-phenylene))bis(propan-2-ol) (14)

[0258] Methyl 2-amino-4-(1-(4-(methoxycarbonyl)-3-aminophenyl)vinyl)benzoate 14e (340 mg, 1.04 mmol) was dissolved in tetrahydrofuran (20 mL), cooled to -78°C, and 3.0 M methylmagnesium bromide solution (10.4 mL, 31.3 mmol) was added dropwise. The mixture was slowly heated to room temperature and stirred for 16 hours.

[0259] The reaction was complete after LCMS analysis. Saturated ammonium chloride (50 mL) was added to the reaction solution and extracted with ethyl acetate (40 mL × 3). The organic phase was washed with saturated sodium chloride solution (30 mL × 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′-(ethylene-1,1-dibis(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-H2O+H] + .

[0261] 1 H NMR (400MHz, DMSO-d6) δ6.98 (d, J=8.0Hz, 2H), 6.55 (d, J=1.6Hz, 2H), 6.42 (dd, J1=1.6Hz, J2=10.4Hz, 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′-(ethylene-1,2-diylbis(2-amino-5,1-phenylene))bis(propan-2-ol) (15)

[0263] Example 16: Preparation of Compound 2,2′-(ethylene-1,1-diylbis(2-amino-1,1-phenylene))bis(propan-2-ol) (16)

[0264] Synthesis routes of compounds 15 and 16:

[0265] Step 1: Preparation of (E)-dimethyl 5,5′-(ethylene-1,2-diyl)bis(2-aminobenzoate) (15a) and 5,5′-(ethylene-1,1-diyl)dimethylbis(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 the atmosphere was purged with nitrogen three times. The reaction mixture was stirred at 120°C for 16 hours.

[0267] The reaction was complete after LCMS analysis and the mixture was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 5:1 to 3:1) to afford a mixture of (E)-dimethyl 5,5′-(ethylene-1,2-diyl)bis(2-aminobenzoate) 15a and 5,5′-(ethylene-1,1-diyl)dimethylbis(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′-(ethylene-1,2-diylbis(2-amino-5,1-phenylene))bis(propan-2-ol) (15) and 2,2′-(ethylene-1,1-diylbis(2-amino-1,1-phenylene))bis(propan-2-ol) (16)

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

[0271] The reaction was complete after LCMS detection. Saturated ammonium chloride (20 mL) was added to the reaction solution and extracted with ethyl acetate (50 mL × 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 (E)-2,2′-(ethylene-1,2-diylbis(2-amino-5,1-phenylene))bis(propan-2-ol) 15 (168 mg, off-white solid) and 2,2′-(ethylene-1,1-diylbis(2-amino-1,1-phenylene))bis(propan-2-ol) 16 (21 mg, off-white solid, total yield: 36.4%).

[0272] The mass spectrum and H NMR spectrum of compound 15 are as follows:

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

[0274] 1 H NMR (400MHz, DMSO-d6) δ7.14-7.12 (m, 4H), 6.74 (s, 2H), 6.57 (d, J=7.6Hz, 2H), 5.50 (br s, 4H), 5.21 (s, 2H), 1.53 (s, 12H).

[0275] The mass spectrum and H NMR spectrum of compound 16 are as follows:

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

[0277] 1 H NMR (400MHz, DMSO-d6) δ6.98 (d, J=1.6Hz, 2H), 6.91 (dd, J1=2.0Hz, J2=8.4Hz, 2H), 6.57 (d, J=7.6Hz, 2H), 5.52 (br s, 4H), 5.22 (s, 2H), 5.20 (s, 2H), 1.45 (s, 12H).

[0278] 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] 5-Hydroxy-2-nitrobenzoic acid (6.0 g, 32.8 mmol) was dissolved in methanol (60 mL), and concentrated sulfuric acid (1 mL) was added dropwise. The mixture was heated to 80°C and stirred for 12 hours under nitrogen protection.

[0281] The reaction was complete after LCMS detection, and the mixture 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 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 5,5′-(ethane-1,2-diylbis(oxy))bis(methyl 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), and 1,2-dibromoethane (1.72 g, 9.14 mmol) and potassium carbonate (1.68 g, 12.2 mmol) were added. The reaction mixture was stirred at 80°C under nitrogen for 12 hours.

[0285] The reaction was complete after LCMS analysis. Water (20 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL × 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:Et3N = 5:1:0.1% to 1:3:0.1%) to afford methyl 5-(2-bromoethoxy)-2-nitrobenzoate (900 mg, yellow oil, yield: 49%) 17b and methyl 5,5′-(ethane-1,2-diylbis(oxy))bis(2-nitrobenzoate) (220 mg, yellow solid, yield: 17%) 17b-1.

[0286] The mass spectrum and H NMR spectrum of compound 17b are as follows:

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

[0288] 1H NMR (300MHz, CDCl3) δ8.04 (d, J=8.7Hz, 1H), 7.09-7.03 (m, 2H), 4.39 (t, J=6.0Hz, 2H), 3.94 (s, 3H), 3.67 (t, J=6.0Hz, 2H).

[0289] The mass spectrum and H NMR spectrum of compound 17b-1 are as follows:

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

[0291] 1 H NMR (300MHz, CDCl3) δ8.05 (d, J=9.0Hz, 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 mixture was stirred at 80°C under nitrogen for 12 hours.

[0294] The reaction was complete after LCMS analysis. Water (20 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL × 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:Et3N = 5:1:0.1% to 1:3:0.1%) to afford 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 (300MHz, CDCl3) δ8.05 (d, J=8.7Hz, 1H), 7.81 (d, J=8.7Hz, 1H), 7.32 (d, J=2.4Hz, 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), 10% palladium on carbon (45 mg) was added, and the mixture was hydrogenated using a hydrogen balloon at room temperature for 12 hours.

[0299] The reaction was complete after LCMS detection. The mixture was filtered through celite and concentrated under reduced pressure to give the crude product methyl 2-amino-4-(2-(4-amino-3-(methoxycarbonyl)phenoxy)ethoxy)benzoate 17d (340 mg, yellow solid, yield: 88%).

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

[0301] 1 H NMR (300MHz, DMSO-d6) δ7.63 (d, J=9.0Hz, 1H), 7.23 (d, J=3.0Hz, 1H), 7.03 (dd, J=9.0Hz, 3.0Hz, 1H), 6.75 (d, J=9.3Hz, 1H), 6.69 (br s, 2H), 6.37-6.31 (m, 3H), 6.17 (dd, J=9.0Hz, 2.4Hz, 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), cooled to -78°C, and 3.0 M methylmagnesium bromide solution (9.4 mL, 28.2 mmol) was added dropwise. The mixture was slowly warmed to room temperature and stirred for 12 hours.

[0304] The reaction was complete after LCMS detection, and water (20 mL) was added to the reaction solution and extracted with ethyl acetate (20 mL×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 = 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 (400MHz, DMSO-d6) δ6.89 (d, J=8.4Hz, 1H), 6.65 (d, J=2.8Hz, 1H), 6.62 (dd, J=8.4Hz, 2.8 Hz, 1H), 6.54 (d, J=8.4Hz, 1H), 6.21 (d, J=2.4Hz, 1H), 6.08 (dd, J=8.4Hz, 2.4Hz, 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] Example 18: Preparation of Compound 2,2′-((Ethane-1,2-diylbis(oxy))bis(6-amino-3,1-phenylene))di(propan-2-ol) trifluoroacetate (18)

[0308] Step 1: Preparation of 5,5′-(ethane-1,2-diylbis(oxy))bis(methyl 2-aminobenzoate) (18a)

[0309] 5,5′-(Ethane-1,2-diylbis(oxy))bis(methyl 2-nitrobenzoate) 17b-1 (220 mg, 0.523 mmol) was dissolved in methanol (5 mL), tetrahydrofuran (5 mL) and ethyl acetate (5 mL), 10% palladium carbon (44 mg) was added, and hydrogenation was carried out using a hydrogen balloon at room temperature for 12 hours.

[0310] The reaction was complete by LCMS, and the mixture was filtered through celite. The filtrate was concentrated under reduced pressure to give 5,5′-(ethane-1,2-diylbis(oxy))bis(methyl 2-aminobenzoate) 18a (200 mg crude product, yellow solid, yield: 100%).

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

[0312] 1 H NMR (300MHz, CDCl3) δ7.42 (d, J=3.0Hz, 2H), 7.01 (dd, J=8.7Hz, 2.7Hz, 2H), 6.63 (d, J=9.0Hz, 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))di(propan-2-ol) trifluoroacetate (18)

[0314] 5,5′-(Ethane-1,2-diylbis(oxy))bis(methyl 2-aminobenzoate) 18a (200 mg crude product, 0.523 mmol) was dissolved in tetrahydrofuran (2 mL), cooled to −78° C. 3.0 M methylmagnesium bromide solution (5.2 mL, 15.6 mmol) was added dropwise, and the mixture was slowly warmed to room temperature and stirred for 12 hours.

[0315] The reaction was complete after LCMS detection, and water (20 mL) was added to the reaction solution and extracted with ethyl acetate (20 mL×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% trifluoroacetic acid aqueous solution = 5% to 35%) to give 2,2′-((ethane-1,2-diylbis(oxy))bis(6-amino-3,1-phenylene))di(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 (400MHz, DMSO-d6) δ9.42 (br s, 4H), 7.20 (d, J=8.8Hz, 2H), 6.97 (dd, J=8.4Hz, 2.8Hz, 2H), 6.93 (d, J=2.4Hz, 2H), 4.32 (s, 4H), 1.55 (s, 12H).

[0318] 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-nitropyridinecarboxylate (19a)

[0320] Dissolve methyl 5-hydroxy-3-nitropyridine carboxylate (3.0 g, 15.1 mmol) in N,N-dimethylformamide (60 mL), add 1,2-dibromoethane (8.54 g, 45.4 mmol) and potassium carbonate (4.17 g, 30.2 mmol), and stir the reaction mixture at 80°C under nitrogen overnight.

[0321] The reaction was complete after LCMS analysis. Water (100 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phase was washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, and 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 afford methyl 5-(2-bromoethoxy)-3-nitropyridinecarboxylate 19a (2.1 g, yellow oil, yield: 46%).

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

[0323] 1 H NMR (300MHz, CDCl3) δ8.56 (d, J=2.4Hz, 1H), 7.70 (d, J=2.7Hz, 1H), 4.47 (t, J=6.0Hz, 2H), 3.99 (s, 3H), 3.70 (t, J=6.0Hz, 1H).

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

[0325] Dissolve methyl 5-(2-bromoethoxy)-3-nitropyridine carboxylate (500 mg, 1.64 mmol) in N,N-dimethylformamide (5 mL), add methyl 5-hydroxy-2-nitrobenzoate (323 mg, 1.64 mmol) and potassium carbonate (679 mg, 4.92 mmol), and stir the reaction mixture at 80°C under nitrogen overnight.

[0326] The reaction was complete after LCMS analysis. 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, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA:Et3N = 2:1:0.1% to 1:5:0.1%) to afford 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)picolinate (19c)

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

[0330] The reaction was completed by LCMS, and the mixture was filtered through celite and 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), cooled to -78°C, and 3.0 M methylmagnesium bromide solution (5.5 mL, 16.5 mmol) was added dropwise. The mixture was slowly warmed to room temperature and stirred for 12 hours.

[0334] The reaction was complete after LCMS analysis. 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, and 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 obtain the crude product, which was further purified by preparative chromatography (acetonitrile:0.1% aqueous ammonium bicarbonate = 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] 1 H NMR (400MHz, DMSO-d6) δ7.44 (d, J=2.8Hz, 1H), 6.65 (d, J=2.4Hz, 1H), 6.62 (dd, J=8.4Hz, 2.8Hz, 1H), 6.58 (d, J=2.4Hz, 1H), 6.54 (d, J=8.8Hz, 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))di(propan-2-ol) (20)

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

[0339] 4-Bromo-2-nitrobenzoic acid (10 g, 40.6 mmol) was dissolved in methanol (100 mL), and concentrated sulfuric acid (1 mL) was added dropwise. The mixture was heated to 80°C and stirred for 12 hours under nitrogen protection.

[0340] The reaction was complete after LCMS analysis. The reaction solution was 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 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 (300MHz, CDCl3) δ 8.02 (d, J=1.8Hz, 1H), 7.80 (dd, J=8.1Hz, 1.5Hz, 1H), 7.65 (d, J=8.1Hz, 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'-bis(1,3,2-dioxaborolane) (4.88 g, 19.2 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (697 mg, 0.953 mmol), and potassium acetate (2.8 g, 28.6 mmol) were added. The reaction mixture was stirred at 80°C under nitrogen for 12 hours.

[0345] The reaction was complete after LCMS detection. The mixture was filtered through celite and the filtrate was concentrated under reduced pressure to give 4-(methoxycarbonyl)-3-nitrophenyl)boronic acid 20b (6.5 g crude product, 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] 4-(Methoxycarbonyl)-3-nitrophenyl)boronic acid 20b (6.5 g crude product, 19.2 mmol) was dissolved in tetrahydrofuran (40 mL), and 30% hydrogen peroxide (10.9 g, 96 mmol) was added. The reaction mixture was stirred at room temperature under nitrogen for 12 hours.

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

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

[0351] 1 H NMR (300MHz, DMSO-d6) δ7.77 (d, J=8.4Hz, 1H), 7.23 (d, J=2.4Hz, 1H), 7.11 (dd, J=8.4Hz, 2.4Hz, 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 product, 9.60 mmol) was dissolved in methanol (30 mL), 10% palladium on carbon (300 mg) was added, and the mixture was hydrogenated using a hydrogen balloon at room temperature for 12 hours.

[0354] The reaction was completed by LCMS detection. The product was filtered through celite and the filter cake was washed with methanol (30 mL×3). 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 4,4′-(ethane-1,2-diylbis(oxy))bis(dimethyl 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 mixture was stirred at 80°C under nitrogen for 12 hours.

[0358] The reaction was complete after LCMS analysis. 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, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA:Et3N = 10:1:0.1% to 1:3:0.1%) to obtain 4,4′-(ethane-1,2-diylbis(oxy))bis(dimethyl 2-aminobenzoate) 20e (380 mg, yellow solid, yield: 50%).

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

[0360] 1 H NMR (300MHz, DMSO-d6) δ7.63 (d, J=9.0Hz, 2H), 6.70 (br s, 4H), 6.31 (d, J=2.4Hz, 2H), 6.18 (dd, J=8.7Hz, 2.4Hz, 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))di(propan-2-ol) (20)

[0362] Dissolve 4,4′-(ethane-1,2-diylbis(oxy))bis(dimethyl 2-aminobenzoate) 20e (380 mg, 1.05 mmol) in tetrahydrofuran (4 mL), cool to -78°C, add 3.0 M methylmagnesium bromide solution (21 mL, 63 mmol) dropwise, and slowly warm to room temperature and stir for 12 hours.

[0363] The reaction was complete after LCMS analysis. The reaction solution 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, and 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 preparative chromatography (acetonitrile:0.1% aqueous ammonium bicarbonate = 20% to 60%) to give 2,2′-((ethane-1,2-diylbis(oxy))bis(2-amino-4,1-phenylene))di(propan-2-ol) 20 (210 mg, white solid, yield: 56%).

[0364] MS (ESI): m / z 325.1 [M-2H2O+H] + .

[0365] 1 H NMR (400MHz, DMSO-d6) δ6.89 (d, J=8.8Hz, 2H), 6.22 (d, J=2.4Hz, 2H), 6.08 (dd, J=8.4Hz, 2.8Hz, 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 21a

[0368] Ethyltriphenylphosphonium bromide (26.75 g, 72 mmol) was dissolved in tetrahydrofuran (200 mL) and the atmosphere was replaced with nitrogen three times. Potassium tert-butoxide (8.1 g, 72 mmol) was added portionwise to the reaction mixture at 0°C. After 30 minutes, methyl 5-formyl-2-nitrobenzoate (10.0 g, 47.8 mmol) was added to the reaction mixture. The reaction mixture was slowly warmed to room temperature and reacted for 3 hours.

[0369] The reaction was complete after LCMS analysis. The reaction solution was diluted with saturated ammonium chloride (100 mL) and extracted with ethyl acetate (100 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 (PE:EA = 15:1) to afford 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, and the reaction mixture was stirred at room temperature for 1 hour.

[0373] The reaction was complete by LCMS. The reaction solution was diluted with dichloromethane (200 mL), washed with saturated sodium bisulfite (30 mL x 3) and saturated sodium chloride (50 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 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) and the reaction was stirred at room temperature for 20 minutes.

[0377] The reaction was complete by LCMS. The reaction solution 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 afford 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), and 1,1′-bis(diphenylphosphino)ferrocenepalladium(II) chloride (420 mg, 0.57 mmol) were dissolved in water (2.0 mL) and 1,4-dioxane (20 mL). The atmosphere was purged with nitrogen three times. The reaction mixture was stirred at 100°C for 3 hours.

[0381] The reaction was complete after LCMS analysis, and the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=20:1 to 10:1) to obtain 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) according to the method of Step 5 of Example 14, with a yield of 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 methylmagnesium bromide solution (29.3 mL, 88 mmol) according to the method of Step 6 of Example 14. Yield: 51.3%.

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

[0389] 1 H NMR (400MHz, MeOD) δ6.98 (s, 1H), 6.90 (s, 1H), 6.86 (d, J = 2.0Hz, 1H), 6.79 (d, J = 8.0Hz, 1H), 6.71 (d, J = 8.0Hz, 1H), 6.58 (d, J=8.0Hz, 1H), 5.89-5.84 (m, 1H), 1.71 (d, J=6.8Hz, 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 methyl 5-bromo-2-nitrobenzoate (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) according to the method of Step 4 of Example 14, with a yield of 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) according to the method of Step 2 of Example 21. 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) according to the method of Step 3 of Example 21, with a yield of 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 methyl 2-amino-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (1.19 g, 4.3 mmol) according to the method of Step 4 of Example 21, with a yield of 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) according to the method of Step 5 of Example 14. 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 methylmagnesium bromide solution (10 mL, 30.0 mmol) according to the method of Step 6 of Example 14. Yield: 52.1%.

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

[0409] 1 H NMR (400MHz, DMSO-d6) δ6.75 (d, J=2.0Hz, 2H), 6.58 (dd, J=1.6Hz, 8.0Hz, 2H), 6.48 (d, J=8.0Hz, 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-1,3,2-dioxaborolan-2-yl)allyl)oxy)silane (9.66 g, 32.37 mmol), potassium carbonate (8.6 g, 62.25 mmol), and 1,1′-bis(diphenylphosphino)ferrocenepalladium(II) dichloride (1.82 g, 2.49 mmol) were dissolved in 1,4-dioxane (60 mL) and water (6 mL), and the atmosphere was purged with nitrogen three times. The reaction mixture was stirred at 90°C for 3 hours.

[0413] The reaction was complete after TLC, and the reaction solution 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 obtain 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), and 1.0 M aqueous hydrochloric acid solution (10 mL) was added, and the mixture was stirred at room temperature for 2 hours.

[0417] The reaction was complete as determined by LCMS, and the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=8:1) to give 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. The reaction solution was slowly warmed to room temperature and stirred for 3 hours.

[0421] The reaction was complete after LCMS analysis. Saturated aqueous sodium bicarbonate was added to the reaction mixture, followed by extraction 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 afford 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 methyl 5-hydroxy-2-nitrobenzoate (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 heated to 80°C and stirred for 3 hours.

[0425] The reaction was complete after LCMS analysis. Water was added to the reaction solution, and the mixture 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 afford 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) according to the method of Step 5 of Example 14. Yield: 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 methylmagnesium bromide solution (9 mL, 28 mmol) according to the method of Step 6 of Example 14. Yield: 11.3%.

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

[0433] 1 H NMR (400MHz, MeOD) δ7.76 (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 = 5.6Hz, 1H), 1.59 (s, 6H) 1.58 (s, 6H).

[0434] In vitro activity evaluation

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

[0436] Main principles of the experiment

[0437] Elevated levels of reactive aldehydes have been found in a variety of inflammatory eye diseases. If these aldehydes are not eliminated promptly, they will accelerate inflammatory symptoms and worsen the eye disease. This experiment simulates the in vivo environment and selects relatively superior compounds based on their ability to complex with aldehydes in the body.

[0438] Experimental materials and reagents

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

[0440] Experimental procedures

[0441] S1. Dissolve sulfobutyl-β-cyclodextrin (3 g) in phosphate buffer (12 ml) to prepare a solution. Use a pipette to pipette 2.88 ml of triolein and 2.88 ml of linoleic acid, respectively, and mix them in a 1:1 ratio.

[0442] S2. To a 2 ml Eppendorf tube at room temperature, 48 μL of the triolein / linoleic acid mixture was added, followed by a solution of the compound prepared in an example of the present invention (10 μmol, 1.0 equivalent) in dimethyl sulfoxide (5 μL). After mixing, 100 μL of the sulfobutyl-β-cyclodextrin solution was added, followed by a solution of nonenal (20 μmol, 2.0 equivalent) in dimethyl sulfoxide (10 μL). The mixture was stirred at 1000 rpm.

[0443] S3, after stirring the reaction for 0 (no nonenal solution was added to the reaction system, and 1510 μL of methanol was added to the reaction system), 10, 20, 40, and 60 minutes, 1500 μL of methanol was added to an Eppendorf tube. After high-speed vortexing, the tube was quickly centrifuged and the solution was transferred to a high-performance liquid chromatography injection for analysis.

[0444] Experimental results

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

[0446] Table 1: Liquid phase gradient conditions

[0447] Among them, the injector temperature is 15°C; the column temperature is 40°C; and the UV-visible light detection channel is shown in Table 2 below:

[0448] Table 2: UV-visible light detection channels of Examples 1-20

[0449] Assuming that the absorption coefficient of the complex product is consistent with that of the test compound, the peak area of ​​the test compound in the T0 sample (the sample without nonenal) is set to 100%. The percentage of the complex product generated is calculated based on the ratio of the peak area of ​​the main product peak to the initial peak area of ​​the test compound at each time point to observe the ability of the compound to capture aldehyde. From the UV-visible light detection channel table, it can be seen that nonenal absorbs weakly at the maximum absorption wavelength of each complex product, and its influence on the content of the complex product can be ignored. The percentage content (%) HPLC data of the complex product of the compound of the present invention are shown in Table 3 below:

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

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

[0452] As can be seen from the above table, the compounds prepared in the examples of the present invention all have very significant ability and speed to complex aldehydes, and can be used to treat or prevent diseases with RASP-mediated pathological characteristics, including but not limited to dry eye, Behçet's disease, Sjögren's syndrome, non-infectious uveitis and allergic conjunctivitis.

[0453] Animal efficacy evaluation

[0454] Example 25: Drug efficacy test in animal model of allergic conjunctivitis

[0455] Study Principle and Objective: C48 / 80 is a mast cell degranulating agent that directly binds to receptors on the surface of mast cells, triggering signaling pathways within the cells. Mast cell degranulation leads to the release of various inflammatory mediators. C48 / 80 and the inflammatory response it triggers may directly damage the conjunctival epithelial barrier. Inflammatory mediators and substances such as proteases released by inflammatory cells can degrade junctional proteins between conjunctival epithelial cells, compromising the integrity of the conjunctival epithelium, making the conjunctiva more susceptible to invasion by external pathogens and exacerbating the inflammatory response within the tissue. Furthermore, the inflammatory process stimulates nerve endings in the conjunctival tissue. Inflammatory mediators such as histamine can activate sensory nerve endings, causing symptoms such as pain and itching. Furthermore, this stimulation of nerve endings leads to the release of neuropeptides such as substance P. Substance P has vasodilatory and proinflammatory effects, further exacerbating conjunctival congestion and inflammation. This study investigated the ameliorative effects of the compound of this invention on a rat conjunctivitis model induced by eye drops of C48 / 80.

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

[0457] Grouping: Female Sprague-Dawley rats were randomly divided into three groups: a normal control group, a model control group, an olopatadine hydrochloride group, and a Compound 23 group. The normal control group did not undergo modeling, while all other groups underwent modeling. Detailed grouping information is shown in Table 4.

[0458] Table 4: List of drug administration and dosage design for each group

[0459] Modeling, drug administration, and evaluation: Rats were randomly divided into groups and anesthetized. The normal control group was given 10 μL / eye of normal saline dripped into the conjunctival sac, while the other groups were given 10 μL / eye of C48 / 80 solution (source: MCE; specification and concentration: 50 mg / tube; batch number: 312920; 200 mg / mL, prepared in 0.9% normal saline) dripped into the conjunctival sac. The eyelids were gently closed for 10 s to prevent the solution from overflowing.

[0460] Ten minutes after stimulation, 15 μL / eye of the test substance was instilled into the conjunctival sac, and the eyelids were gently closed for 10 seconds to prevent overflow of the solution. The normal and model control groups were given an equal volume of normal saline. Twenty minutes after the third administration and 24 hours after modeling, ocular examinations were performed under a slit lamp, and clinical scores were performed on both eyes of each animal according to the scoring criteria in Table 5.

[0461] Table 5: Scoring criteria for rat conjunctivitis

[0462] According to the scoring criteria in Table 5, the score values ​​were positively correlated with the severity of ocular inflammation. The experimental results are shown in Figure 1, demonstrating that Compound 23 of the present invention has a significant therapeutic effect on C48 / 80-induced allergic conjunctivitis in SD rats. Figure 1 represents the normal control group, 2 the model control group, 3 the olopatadine hydrochloride group, and 4 the Compound 23 group.

[0463] Although the present invention has been described in detail through the above preferred embodiments, it should be understood that the above description is not intended to limit the present invention. After reading the above description, various modifications and substitutions of the present invention will become apparent to those skilled in the art. Therefore, the scope of protection of the present invention should be defined by the appended claims.

Claims

1. An active aldehyde inhibitor, characterized in that: It comprises a compound represented by general formula (I) or a derivative thereof, in, R 1 is one or more substituents selected from hydrogen, hydroxy, cyano, nitro, halogen, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Alkylcarbonyl and C 1-6 At least one of alkylamino groups; X and Z are independently selected from C or N; Y is selected from -CH=CH-, -C≡C-, Any one of -OCH2CH2O- or -CH=CHCH2O-; 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 Selected from hydrogen, hydroxy, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 At least one of alkylamino groups.

3. The active aldehyde inhibitor according to claim 1, wherein The compound represented by the general formula (I) is selected from: At least one of .

4. A pharmaceutical composition, characterized in that The invention comprises the compound represented by the general formula (I) or a derivative thereof according to any one of claims 1 to 3 as an active ingredient.

5. The pharmaceutical composition according to claim 4, wherein The pharmaceutical composition further comprises pharmaceutical excipients.

6. The pharmaceutical composition according to claim 4, wherein The pharmaceutical composition is a detection reagent or kit, which contains 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 represented by general formula (I) and is used to identify the RASP ligand.

7. A use of the active aldehyde inhibitor according to any one of claims 1 to 3, characterized in that: Used in the preparation of drugs as aldehyde trapping agents.

8. The use according to claim 7, characterized in that The medicine is used for treating or preventing various diseases caused by active aldehyde substances.

9. The use according to claim 8, characterized in that The diseases are eye diseases.

10. The use according to claim 9, characterized in that The eye disease comprises at least one of dry eye, allergic conjunctivitis, Behcet's disease, Sjögren's syndrome and uveitis.

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