Small-molecule active aldehyde scavenger compound and application thereof

Abstract

The invention relates to the field of medicines, in particular to a small-molecule active aldehyde scavenger compound and application thereof. The compound comprises a compound with the following structure or a derivative thereof. The compound is further prepared into an ophthalmic preparation, so that the residence time of the medicine in the cornea is prolonged, the efficiency of removing active aldehyde in vitro is improved, and the expression of a human corneal epithelial cell inflammatory factor IL-6 is remarkably inhibited. The technical defects that a traditional xerophthalmia medicine is short in effect and single in mechanism are overcome, and the obvious clinical transformation value is achieved.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, specifically to a small molecule active aldehyde scavenger compound and its application. Background Technology

[0002] Dry eye disease (DED) is a chronic ocular surface disease caused by insufficient tear secretion, excessive evaporation, or abnormal tear composition. The prevalence of DED in China is as high as 32.1%, and 5%–50% of people over 50 years old in Asia suffer from dry eye symptoms. DED has become a global research focus. Its core pathological mechanism is a cascade of inflammatory responses (abnormally elevated levels of inflammatory factors such as IL-6 and TNF-α) and oxidative stress (accumulation of reactive aldehydes such as 4-hydroxynonenal) triggered by tear film homeostasis imbalance, severely impacting patients' quality of life and potentially leading to visual impairment. Current mainstream treatments have significant drawbacks. For example, cyclosporine A has a slow onset of action and is intolerant to some patients, causing a burning sensation after administration; long-term use of glucocorticoids can easily lead to increased intraocular pressure; and artificial tears cannot fundamentally treat the inflammatory pathways. Clinically, there is an urgent need for novel treatment strategies that combine rapid onset of action, long-term maintenance, and multi-mechanism intervention.

[0003] Reproxalap (NS-2), a novel active aldehyde small molecule inhibitor, exerts its anti-inflammatory effect through covalent binding with RASP. Preclinical studies have shown that it can rapidly inhibit the release of inflammatory mediators in tears. However, it has drawbacks such as short ocular retention time, limited corneal penetration, the need for frequent administration, and significant ocular irritation. Summary of the Invention

[0004] To address the shortcomings and deficiencies of existing technologies, this invention provides a small-molecule active aldehyde scavenger compound with good stability. The specific details of this invention are as follows: In a first aspect, the present invention provides a small molecule active aldehyde scavenger compound, said compound comprising a compound or a derivative thereof with the following structure: ; R1 includes hydrogen and C. 1-3 Alkyl, C 2-3 alkenyl, C 2-3 alkynyl group, C 1-3 Alkoxy, C 3-5 cycloalkyl, C 3-5 Cycloalkyloxy, halogen, nitro, cyano, trifluoromethyl, 3-7 membered heterocyclic groups, unsubstituted or Ra-substituted C 6-10 aryl, unsubstituted or Ra-substituted C 6-10 Aromatic heteroalkyl groups. Each Ra group may be identical or different, and is independently selected from halogens, C... 1-5 Alkyl, C1-5 Alkoxy, C 6-10 At least one of aryl acyl groups.

[0005] Preferably, R1 is a halogen or C 1-3 At least one of alkoxy, trifluoromethyl, nitro, and cyano groups; Wherein R2 includes at least one of the following structures: .

[0006] Furthermore, the general structural formula of the connector is as follows, where X is selected from carbon atoms, nitrogen atoms, and oxygen atoms, and n is 0~3; .

[0007] Furthermore, the derivatives of the compound include at least one of the following: enantiomers, diastereomers, geometric isomers, free forms, and pharmaceutically acceptable salts, hydrates, solvates, and esters.

[0008] In a second aspect, the present invention provides a composition comprising the aforementioned compound and pharmaceutically acceptable additives.

[0009] Further, the additives include at least one of the following: binders, fillers, diluents, disintegrants, suspending agents, suspending aids, sustained-release agents, controlled-release agents, lyophilization protectants, coating agents, enteric materials, lubricants, flow aids, anti-adhesion agents, sweeteners, flavoring agents, plasticizers, opacifiers, solubilizers, humectants, solvents, osmotic pressure regulators, colorants, pigments, surfactants, emulsifiers, water-soluble matrices, fat-soluble matrices, oleic matrices, pore-forming agents, gelling agents, preservatives, buffers, chelating agents, and antioxidants.

[0010] A third aspect of the invention provides the use of the said compound or the said composition in any of the following aspects: A1. Application in the preparation of products that promote tear secretion; A2. Application in the preparation of products that promote corneal damage repair; A3. Applications in preparing products with improved crystal morphology; A4. Applications in the preparation of products that inhibit the expression of pro-inflammatory factors; A5. Application in the preparation of products that promote epithelial proliferation and accelerate corneal structural reconstruction.

[0011] In a fourth aspect, the present invention provides the use of the said compound or the said composition in the preparation of medicaments for treating and / or preventing eye diseases, cataracts, keratoconus, Fuch's endothelial dystrophy in the cornea, uveitis, allergic conjunctivitis, ocular pemphigoid scarring, and refractive keratotomy (PRK) healing disorders.

[0012] In a fifth aspect, the present invention provides the use of the said compound or the said composition in the preparation of medicaments for treating and / or preventing corneal healing-related diseases, diseases related to tear lipid degradation or lacrimal gland dysfunction, and inflammatory eye diseases.

[0013] In a sixth aspect, the present invention provides the use of the said compound or the said composition in the preparation of medicaments for treating and / or preventing eye diseases.

[0014] Furthermore, the drug is in the form of at least one of solid, liquid or gas.

[0015] The beneficial effects of the present invention include, but are not limited to: This compound, when further formulated into an ophthalmic preparation, increases the drug's residence time in the cornea, enhances its efficiency in clearing reactive aldehydes in vitro, and significantly inhibits the expression of the inflammatory factor IL-6 in human corneal epithelial cells. This overcomes the technical limitations of traditional dry eye drugs, such as short-lived action and simple mechanisms of action, and possesses clear clinical translational value. Attached Figure Description

[0016] The accompanying drawings, which are included to provide a further understanding of the invention and form part of this invention, illustrate exemplary embodiments of the invention and are used to explain the invention, but do not constitute an undue limitation of the invention. In the drawings: Figure 1 This is the 1H NMR DMSO-d6 303K AV-300 proton NMR spectrum of target compound 15 in this embodiment of the invention; Figure 2 This is the 1H NMR DMSO-d6 303K AV-300 proton NMR spectrum of target compound 16 in this embodiment of the invention; Figure 3 This is the 1H NMR DMSO-d6 303K AV-300 proton NMR spectrum of target compound 28a in this embodiment of the invention; Figure 4 This is the 1H NMR DMSO-d6 303K AV-300 proton NMR spectrum of target compound 29a in this embodiment of the invention; Figure 5This is the 1H NMR DMSO-d6 303K AV-300 proton NMR spectrum of target compound 28b in this embodiment of the invention; Figure 6 This is the 1H NMR DMSO-d6 303K AV-300 proton NMR spectrum of target compound 29b in this embodiment of the invention; Figure 7 This is a schematic diagram showing the stability results of the target compound in physiological saline for 72 h in an embodiment of the present invention; Figure 8 This is a schematic diagram showing the stability results of the target compound in sodium hyaluronate eye drops for 72 hours in an embodiment of the present invention; Figure 9 This is a schematic diagram showing the ability of compounds 15, 16, 28a-b and the positive control drug NS-2 to scavenge active aldehydes in the embodiments of the present invention; Figure 10 This is a schematic diagram showing the effect of compound 15 on tear secretion in mice with dry eye syndrome in an embodiment of the present invention; Figure 11 This is a schematic diagram showing the effect of compound 15 on the corneal damage repair level in mice with dry eye syndrome in an embodiment of the present invention; Figure 12 This is a schematic diagram showing the effects of each of the compounds 15 in this invention on the recovery of tear crystallization in mice with dry eye. Figure 13 This is a schematic diagram showing the effect of compound 15 on the levels of inflammatory factors in mice with dry eye syndrome in an embodiment of the present invention. In the diagram, Figure A shows the results of TNF-α and Figure B shows the results of IL-1β. Detailed Implementation

[0017] The present invention is described in detail below with reference to the embodiments, but the present invention is not limited to these embodiments. Unless otherwise specified, the raw materials and catalysts in the embodiments of the present invention are all purchased through commercial channels.

[0018] Example 1: Synthesis of tert-butyl (8-aminooctyl)carbamate (YY-1)

[0019] 1,8-Octadiamine (5 g, 34.68 mmol, 1.0 equiv) was dissolved in anhydrous dichloromethane and stirred until dissolved. Et3N (4.0 equiv) was then added, and the mixture was covered with a constant-pressure dropping funnel. Di-tert-butyl dicarbonate (0.6 equiv) was dissolved in anhydrous dichloromethane and added to the constant-pressure dropping funnel. Nitrogen gas was purged, and di-tert-butyl dicarbonate was added dropwise. The mixture was stirred overnight at room temperature. The reaction was monitored by TLC (DCM:MeOH = 6:1) and developed with alkaline KMnO4. After the starting material had reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 6:1) to give intermediate YY-1 (3.5 g). The yield was 41%, and the product was a colorless oil.

[0020] Example 2: Synthesis of tert-butyl (8-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)octyl)carbamate (YY-2)

[0021] Intermediate YY-1 (3.5 g, 14.33 mmol, 1.0 equiv) was dissolved in anhydrous dichloromethane, and Et3N (1.5 equiv) was added. After pre-cooling at 0 °C for 15 min, maleic anhydride (1.0 equiv) dissolved in anhydrous dichloromethane was added dropwise, purging with nitrogen, and the mixture was stirred at room temperature for 4 h. After the starting material reacted completely, the mixture was concentrated, dissolved in acetone, and Et3N (1.5 equiv) and acetic anhydride (1.5 equiv) were added. The mixture was then heated to 60 °C under nitrogen protection and stirred for 20 h. The reaction was monitored by TLC (DCM:MeOH = 8:1), and the reaction was developed with basic KMnO4. After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 8:1) to obtain intermediate YY-2 (1.6 g). The yield was 35%, and the product was a white solid powder.

[0022] Example 3: Synthesis of 1-(8-aminooctyl)-1H-pyrrole-2,5-dione (YY-3)

[0023] Intermediate YY-2 (1.6 g, 4.97 mmol, 1.0 equiv) was dissolved in ethyl acetate, and a hydrogen chloride-ethyl acetate solution (2 M in ethyl acetate, 4 equiv) was added. Nitrogen gas was then introduced, and the mixture was stirred at room temperature for 6 h. The reaction was monitored by TLC (DCM: MeOH = 10:1), and a colorimetric reaction was performed with basic KMnO4. After the starting material had reacted completely, the reaction mixture was filtered and concentrated to give intermediate YY-3 (1.3 g). The yield was 98%, and the product was a white solid powder.

[0024] Example 4: Synthesis of 4-((8-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)octyl)amino)-4-oxobutyric acid (13)

[0025] Intermediate YY-3 (1.3 g, 4.87 mmol, 1.0 equiv) was dissolved in anhydrous DMF, and succinic anhydride (1.5 equiv) and N,N-diisopropylethylamine (2.0 equiv) dissolved in anhydrous DMF were slowly added dropwise, purging with nitrogen, and stirred overnight at room temperature. The reaction was monitored by TLC (DCM:MeOH = 10:1) with color development on alkaline KMnO4. After the starting material had reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 10:1) to give linker compound 13 (1.31 g). Yield: 83%. White solid powder.

[0026] Example 5: Synthesis of tert-butyl 2-(2-(2-aminoethoxy)ethoxy)ethylcarbamate (L-1)

[0027] 1,8-Diamino-3,6-dioxane (5 g, 33.76 mmol, 1.0 equiv) was dissolved in anhydrous dichloromethane and stirred until dissolved. Et3N (4.0 equiv) was then added, and the mixture was covered with a constant-pressure dropping funnel. Di-tert-butyl dicarbonate (0.6 equiv) was dissolved in anhydrous dichloromethane and added to the constant-pressure dropping funnel. Nitrogen gas was purged, and di-tert-butyl dicarbonate was added dropwise. The mixture was stirred overnight at room temperature. The reaction was monitored by TLC (DCM:MeOH = 6:1) with color development on alkaline KMnO4. After the starting material had reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 6:1) to give intermediate L-1 (3.6 g). Yield: 43%, white solid powder.

[0028] Example 6: Synthesis of tert-butyl (2-(2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)ethoxy)ethoxy)ethyl)carbamate (L-2)

[0029] Intermediate L-1 (3.6 g, 14.50 mmol, 1.0 equiv) was dissolved in anhydrous dichloromethane, and Et3N (1.5 equiv) was added. After pre-cooling at 0 °C for 15 min, maleic anhydride (1.0 equiv) dissolved in anhydrous dichloromethane was added dropwise, purging with nitrogen, and the mixture was stirred at room temperature for 4 h. After the starting material reacted completely, the mixture was concentrated, dissolved in acetone, and Et3N (1.5 equiv) and acetic anhydride (1.5 equiv) were added. The mixture was then heated to 60 °C under nitrogen protection and stirred for 20 h. The reaction was monitored by TLC (DCM:MeOH = 8:1), and the reaction was observed with basic KMnO4. After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 8:1) to give intermediate L-2 (1.5 g). The yield was 32%, and the product was a milky white oil.

[0030] Example 7: Synthesis of 1-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-1H-pyrrole-2,5-dione (L-3)

[0031] Intermediate L-2 (1.5 g, 4.57 mmol, 1.0 equiv) was dissolved in ethyl acetate, and a hydrogen chloride-ethyl acetate solution (2 M in ethyl acetate, 4 equiv) was added. Nitrogen gas was then introduced, and the mixture was stirred at room temperature for 6 h. The reaction was monitored by TLC (DCM: MeOH = 10:1), and a colorimetric reaction was performed with basic KMnO4. After the starting material had reacted completely, the reaction mixture was filtered and concentrated to give intermediate L-3 (1.16 g). The yield was 96%, and the product was a yellow oil.

[0032] Example 8: Synthesis of 4-((2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)ethoxy)ethoxy)ethyl)amino)-4-oxobutyric acid (14)

[0033] Intermediate L-3 (1.16 g, 4.39 mmol, 1.0 equiv) was dissolved in anhydrous DMF, and succinic anhydride (1.5 equiv) and N,N-diisopropylethylamine (2.0 equiv) dissolved in anhydrous DMF were slowly added dropwise, purging with nitrogen, and stirred overnight at room temperature. The reaction was monitored by TLC (DCM:MeOH = 10:1) with color development on alkaline KMnO4. After the starting material had reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 10:1) to give linker compound 14 (864 mg). Yield: 60%. Pale yellow oil.

[0034] Example 9: Synthesis of 1-bromo-3-hydroxy-3-methyl-2-butanone (2)

[0035] Preparation of NaHSO4·SiO2 catalyst. SiO2 (5 g, 83.22 mmol, 1.0 equiv) was added to an aqueous solution of NaHSO4·H2O (11.49 g, 83.22 mmol, 1.0 equiv), and the reaction mixture was stirred for 15 min. The mixture was then slowly heated until a white solid formed. Finally, the solid was dried in an oven at 120 °C for 48 h to obtain the NaHSO4·SiO2 catalyst. Yield: 99%. White solid.

[0036] Starting material 1 (3-hydroxy-3-methyl-2-butanone) (2.00 g, 19.58 mmol, 1.0 equiv) was dissolved in anhydrous CCl4 (100 mL), and N-bromosuccinimide (NBS) (4.18 g, 23.50 mmol, 1.2 equiv) was added. The mixture was stirred and NaHSO4·SiO2 (1.76 g, 9.79 mmol, 0.5 equiv) was added. The mixture was then heated to 80 °C under reflux and nitrogen protection, and stirred for 2 h. The reaction was monitored by TLC (DCM:MeOH = 100:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 100:1) to give intermediate 2 (2.63 g). Yield: 75%. Pale yellow oil.

[0037] Example 10: Synthesis of 1-(3-hydroxy-3-methyl-2-oxobutyl)pyridine-1-onium (3)

[0038] Intermediate 2 (2.0 g, 11.11 mmol, 1.0 equiv) was dissolved in anhydrous ethanol and pre-cooled at 0 °C for 15 min. Pyridine (1.05 g, 13.33 mmol, 1.2 equiv) was then added, and stirring continued for 15 min. The mixture was then heated to 65 °C under nitrogen protection and stirred overnight. The reaction was monitored by TLC (DCM: MeOH = 10:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM: MeOH = 10:1) to give intermediate 3 (2.36 g). Yield: 82%. A pale yellow solid powder. Results are as follows: Figure 2 As shown, 1 H NMR (300 MHz, DMSO- d 6) δ 9.00 – 8.89 (m,2H), 8.69 (tt, J = 7.8, 1.4 Hz, 1H), 8.31 – 8.17 (m, 2H), 6.13 (s, 2H), 5.89 (s, 1H), 1.35 (s, 6H).

[0039] Example 11: Synthesis of 2-amino-5-iodobenzyl alcohol (5)

[0040] 2-Amino-5-iodobenzoic acid (15 g, 57.03 mmol, 1.0 equiv) was dissolved in anhydrous tetrahydrofuran. Lithium aluminum hydride (1M solution in THF, 1.2 equiv) was added to a constant-pressure dropping funnel, and nitrogen was purged. After pre-cooling at 0 °C for 15 min, lithium aluminum hydride was added dropwise. The mixture was stirred at room temperature for 6 h, and the reaction was monitored by TLC (PE:EA = 2:1). After the reactants had reacted completely, 1 mL of water was added dropwise per gram of lithium aluminum hydride at 0 °C, followed by 1 mL of 15% sodium hydroxide solution, and finally 3 mL of water. The mixture was brought to room temperature and stirred for 30 min. Then, an appropriate amount of anhydrous sodium sulfate was added, and the mixture was stirred for another 30 min. The mixture was filtered, and the filter cake was washed 2 to 3 times with dichloromethane. The organic phases were combined and extracted with saturated brine (3 × 100 mL). The extract was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 2:1). 1) Intermediate 5 (8.5 g) was obtained. Yield: 60%. Pale yellow solid powder.

[0041] Example 12: Synthesis of (2-amino-5-((trimethylsilyl)ethynyl)phenyl)methanol (6)

[0042] Intermediate 5 (8.5 g, 34.14 mmol, 1.0 equiv), triphenylphosphine (0.02 equiv), bis-(triphenylphosphine)-dichloropalladium (0.01 equiv), and CuI (0.03 equiv) were dissolved in a DMF / Et3N solution that had been degassed by bubbling. Nitrogen gas was then introduced, followed by the addition of trimethylsilylacetylene (1.7 equiv). The reaction was stirred at room temperature for 22 h, and the reaction was monitored by TLC (PE:EA = 2:1). After the starting material had completely reacted, the reaction system was concentrated by vacuum distillation. The residue was dissolved in dichloromethane and washed with 1 M hydrochloric acid solution. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (PE:EA = 2:1) to obtain intermediate 6 (5.6 g). Yield: 75%. A pale yellow solid powder. Results are as follows: Figure 3 As shown, 1 H NMR (400 MHz, DMSO- d 6) δ 7.17 (d, J = 2.1 Hz, 1H), 7.05 (dd, J = 8.2, 2.1 Hz, 1H), 6.55 (d, J = 8.2 Hz, 1H),5.37 (s, 2H), 5.06 (t, J = 5.5 Hz, 1H), 4.34 (d, J = 5.5 Hz, 2H), 0.19 (s, 9H).

[0043] Example 13, (2-amino-5-((trimethylsilyl)ethynyl)benzaldehyde (7)

[0044] Intermediate 6 (5.6 g, 25.57 mmol, 1.0 equiv) was dissolved in ethyl acetate and pre-cooled at 0 °C for 15 min. Then, 2-iodobenzoic acid (3.0 equiv) was added, and stirring was continued for 15 min. The mixture was then heated to 80 °C under nitrogen protection and stirred for 4 h. The reaction was monitored by TLC (PE:EA = 3:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (PE:EA = 3:1) to give intermediate 7 (3.66 g). Yield: 66%. A pale yellow solid powder. Results are as follows: Figure 4 As shown, 1 H NMR (300 MHz, CDCl3) δ9.84 (d, J = 0.6 Hz, 1H), 7.67 (d, J =2.0 Hz, 1H), 7.41 (dd, J = 8.6, 2.0 Hz, 1H), 6.60 (d, J = 8.6 Hz, 1H), 6.31 (s, 2H), 0.26 (s, 9H).

[0045] Example 14: Synthesis of 1-(2-(2-hydroxypropyl-2-yl)-6-((trimethylsilyl)ethynyl)quinoline-3-yl)pyridine-1-onium (8)

[0046] Intermediate 7 (1.98 g, 9.11 mmol, 1.0 equiv) and intermediate 3 (2.36 g, 9.11 mmol, 1.0 equiv) were dissolved in anhydrous ethanol and pre-cooled at 0 °C for 15 min. Then, intermediate 3 (1.0 equiv) and pyridine (1.0 equiv) were added, and stirring was continued for another 15 min. The mixture was then heated to 85 °C under nitrogen protection and stirred for 36 h. The reaction was monitored by TLC (DCM: MeOH = 10:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM: MeOH = 10:1) to obtain intermediate 8 (2.74 g). Yield: 69%. Yellow solid powder. Results are as follows. Figure 5 As shown, 1 H NMR (400 MHz, DMSO- d 6) δ 9.41 – 9.30 (m, 2H), 8.83 (m, 1H), 8.78 – 8.74(m, 1H), 8.32 – 8.27 (m, 2H), 8.25 (d, J = 1.9 Hz, 1H), 8.11 (dd, J = 8.8, 0.8Hz, 1H), 7.95 (dd, J = 8.8, 1.9 Hz, 1H), 5.27 (s, 1H), 1.58 (s, 6H), 0.29 (s, 9H).

[0047] Example 15: Synthesis of 2-(3-amino-6-((trimethylsilyl)ethynyl)quinoline-2-yl)prop-2-ol (9)

[0048] Intermediate 8 (2.74 g, 6.24 mmol, 1.0 equiv) was dissolved in anhydrous ethanol, and morpholine (2.8 equiv) was added. The mixture was then heated to 85 °C under nitrogen protection and stirred overnight. The reaction was monitored by TLC (PE:EA = 2:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (PE:EA = 2:1) to give intermediate 9 (685 mg). Yield: 37%. Yellow solid powder. Results are as follows. Figure 6 As shown, 1 H NMR (300 MHz, CDCl3) δ 7.85(d, J = 8.6 Hz, 1H), 7.74 (d, J = 1.7 Hz, 1H), 7.48 (dd, J = 8.6, 1.8 Hz, 1H), 7.16 (s, 1H), 4.61 (s, 2H), 1.77 (s, 6H), 0.30 (s, 9H).

[0049] Example 16: Synthesis of 2-(3-amino-6-ynylquinoline-2-yl)prop-2-ol (10)

[0050] Intermediate 9 (685 mg, 2.30 mmol, 1.0 equiv) was dissolved in anhydrous methanol, and tetrabutylammonium fluoride (1 M solution in THF, 1.2 equiv) was added. The mixture was stirred at room temperature for 2 h under nitrogen protection. The reaction was monitored by TLC (PE:EA = 2:1). After the starting material reacted completely, saturated ammonium chloride was added to terminate the reaction. The reaction mixture was filtered, concentrated, dissolved in dichloromethane, extracted with saturated brine, dried over anhydrous sodium sulfate, concentrated in the organic phase, and purified by column chromatography (PE:EA = 2:1) to give intermediate 10 (442 mg). Yield: 85%. Yellow oil. Results are as follows. Figure 7 As shown, 1 H NMR (400MHz, CDCl3) δ 7.85 (d, J = 8.6 Hz, 1H), 7.76 (d, J = 1.8 Hz, 1H), 7.49 (dd, J= 8.6,1.8 Hz, 1H), 7.16 (s, 1H), 4.66 (s, 2H), 3.94 (s, 1H), 3.16 (s, 1H), 1.77 (s, 6H).

[0051] Example 17: Synthesis of 2-(3-amino-6-[1-(2-(2-aminoethoxy)ethyl]-1H-1,2,3-triazol-4-yl)quinoline-2-yl)prop-2-ol (12)

[0052] Intermediate 10 (442 mg, 1.96 mmol, 1.0 equiv) was dissolved in anhydrous tetrahydrofuran, and azido-PEG1-amine (1.1 equiv) and copper(I) 3-methylsalicylate (0.1 equiv) were added. The mixture was stirred at room temperature for 3 h under nitrogen protection. The reaction was monitored by TLC (DCM:MeOH = 5:1). After the starting material was completely reacted, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 5:1) to give intermediate 12 (557 mg). Yield: 80%. Yellow oil. Results are as follows. Figure 1 As shown, 1 H NMR (400 MHz, DMSO- d 6) δ 8.67 (s, 1H), 8.06 (d, J = 1.7 Hz, 1H), 7.85 – 7.67(m, 2H), 7.27 (s, 1H), 5.97 (s, 2H), 5.77 (s, 1H), 4.60 (t, J = 5.2 Hz, 2H), 3.86 (t, J = 5.2 Hz, 2H), 3.40 (t, J = 5.7 Hz, 2H), 2.64 (t, J = 5.7 Hz, 2H), 1.63 (s, 6H).

[0053] Example 18. Synthesis of N1-(2-(2-(4-(3-amino-2-(2-hydroxypropen-2-yl)quinoline-6-yl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl)-N4-(8-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)octyl)succinamide (15)

[0054] Linker compound 13 (200 mg, 0.62 mmol, 1.2 equiv) was dissolved in anhydrous dichloromethane, and then... O -(benzotriazol-1-yl)-N,N,N',N'-tetramethylurea tetrafluoroborate (TBTU, 2 equiv) and Et3N (2 equiv) were dissolved by stirring. After the solution became clear, intermediate 12 (183 mg, 0.51 mmol, 1.0 equiv) was added, and nitrogen was purged. The mixture was stirred at room temperature for 5 h. The reaction was monitored by TLC (DCM: MeOH = 20:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM: MeOH = 20:1) to give target compound 15 (95 mg). Yield: 28%. The product was a pale yellow solid powder. Results are as follows: Figure 2 As shown, 1 H NMR (400 MHz, DMSO- d 6) δ 8.63 (s, 1H), 8.05 (d, J =1.7 Hz, 1H), 7.91 (t, J = 5.7 Hz, 1H), 7.83 – 7.77 (m, 2H), 7.77 – 7.73 (m,1H), 7.27 (s, 1H), 7.00 (s, 2H), 5.96 (s, 2H), 5.75 (s, 1H), 4.59 (t, J = 5.1Hz, 2H), 3.86 (t, J = 5.1 Hz, 2H), 3.45 (t, J = 5.8 Hz, 2H), 3.38 (d, J = 7.0Hz, 2H), 3.20 (q, J = 5.4 Hz, 2H), 2.98 (q, J = 6.6 Hz, 2H), 2.28 (m, 4H), 2.00 (m, 2H), 1.62 (s, 6H), 1.51 – 1.27 (m, 10H).

[0055] Example 19. Synthesis of N1-(2-(2-(4-(3-amino-2-(2-hydroxypropen-2-yl)quinoline-6-yl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl)-N4-(2-(2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)ethoxy)ethoxy)ethyl)succinamide (16)

[0056] Linker compound 14 (206 mg, 0.63 mmol, 1.2 equiv) was dissolved in anhydrous dichloromethane. O-(benzotriazol-1-yl)-N,N,N',N'-tetramethylurea tetrafluoroborate (TBTU, 2 equiv) and Et3N (2 equiv) were added, and the mixture was stirred until dissolved. After the solution became clear, intermediate 12 (186 mg, 0.52 mmol, 1.0 equiv) was added, and the mixture was purged with nitrogen and stirred at room temperature for 5 h. The reaction was monitored by TLC (DCM:MeOH = 20:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 20:1) to give target compound 16 (87 mg). Yield: 25%. A pale yellow solid powder. Results are as follows: Figure 3 As shown, 1 H NMR (600 MHz, DMSO- d 6) δ 8.65 (s, 1H), 8.05 (dd, J =4.5, 1.8 Hz, 1H), 7.94 (t, J = 5.7 Hz, 1H), 7.89 – 7.78 (m, 2H), 7.77 (d, J =8.6 Hz, 1H), 7.36 – 7.17 (m, 2H), 7.12 – 6.58 (m, 1H), 5.96 (d, J = 5.1 Hz, 2H), 5.76 (d, J = 3.5 Hz, 1H), 4.60 (t, J = 5.1 Hz, 2H), 3.87 – 3.85 (m, 2H), 3.58 – 3.49 (m, 2H), 3.48 – 3.40 (m, 4H), 3.36 – 3.30 (m, 4H), 3.22 – 3.17(m, 2H), 2.30 (s, 2H), 2.10 – 1.89 (m, 4H), 1.76 (s, 2H), 1.63 (s, 6H).

[0057] Example 20: Synthesis of (2-amino-4-fluoro-5-iodophenyl)methanol (18)

[0058] 2-Amino-4-fluoro-5-iodobenzoic acid (15 g, 57.03 mmol, 1.0 equiv) was dissolved in anhydrous tetrahydrofuran. Lithium aluminum hydride (1M solution in THF, 1.2 equiv) was added to a constant-pressure dropping funnel, and nitrogen was purged. After pre-cooling at 0 °C for 15 min, lithium aluminum hydride was added dropwise. The mixture was stirred at room temperature for 6 h, and the reaction was monitored by TLC (PE:EA = 2:1). After the reactants had reacted completely, 1 mL of water was added dropwise per gram of lithium aluminum hydride at 0 °C, followed by 1 mL of 15% sodium hydroxide solution, and finally 3 mL of water. The mixture was brought to room temperature and stirred for 30 min. Then, an appropriate amount of anhydrous sodium sulfate was added, and the mixture was stirred for another 30 min. The mixture was filtered, and the filter cake was washed 2 to 3 times with dichloromethane. The organic phases were combined and extracted with saturated brine (3 × 100 mL). The extract was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 2:1). 2:1) Intermediate 18 (8 g) was obtained. Yield: 60%. Pale yellow solid powder.

[0059] Example 21: Synthesis of (2-amino-4-fluoro-5-((trimethylsilyl)ethynyl)phenyl)methanol (19)

[0060] Intermediate 18 (7.8 g, 29.22 mmol, 1.0 equiv), triphenylphosphine (0.02 equiv), bis-(triphenylphosphine)-dichloropalladium (0.01 equiv), and CuI (0.03 equiv) were dissolved in a DMF / Et3N solution that had been degassed by bubbling. Nitrogen gas was then introduced, followed by the addition of trimethylsilylacetylene (1.7 equiv). The reaction was stirred at room temperature for 22 h, and the reaction was monitored by TLC (PE:EA = 2:1). After the reactants had completely reacted, the reaction system was concentrated by vacuum distillation. The residue was dissolved in dichloromethane and washed with 1M hydrochloric acid solution. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (PE:EA = 2:1) to obtain intermediate 19 (4.0 g). Yield: 58%. A pale yellow solid powder was obtained.

[0061] Example 22: Synthesis of 2-amino-4-methoxybenzaldehyde (21)

[0062] The starting material 4-methoxy-2-nitrobenzaldehyde (15 g, 82.86 mmol, 1.0 equiv) was dissolved in anhydrous methanol. Palladium on carbon (1.5 g, 8.29 mmol, 0.1 equiv) and Et3N (16.8 g, 165.71 mmol, 2.0 equiv) were added. Nitrogen gas was first purged, then hydrogen gas was rapidly purged (using two balloons) at 0 °C. The reaction was allowed to proceed at room temperature for 18 h. The reaction was monitored by TLC (PE:EA = 2:1). After the starting material had completely reacted, the mixture was filtered through diatomaceous earth, concentrated, and purified by column chromatography (PE:EA = 2:1) to obtain intermediate 21 (10.6 g). The yield was 85%. It was a pale yellow solid powder.

[0063] Example 23: Synthesis of 2-amino-5-iodo-4-methoxybenzaldehyde (22)

[0064] Intermediate 21 (10 g, 66.20 mmol, 1.0 equiv) was dissolved in anhydrous dichloromethane, and N-iodosuccinimide (14.9 g, 66.20 mmol, 1.0 equiv) was added. The mixture was stirred at room temperature for 1 h, and the reaction was monitored by TLC (PE:EA = 2:1). After the starting material reacted completely, the reaction system was concentrated by vacuum distillation. The residue was dissolved in ethyl acetate, washed three times with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 2:1) to obtain intermediate 22 (16.1 g). Yield: 88%. A pale yellow solid powder.

[0065] Example 24: Synthesis of 2-amino-4-fluoro-5-((trimethylsilyl)ethynyl)benzaldehyde (23a)

[0066] Intermediate 19 (4.0 g, 16.87 mmol, 1.0 equiv) was dissolved in ethyl acetate and pre-cooled at 0 °C for 15 min. Then, 2-iodobenzoic acid (3.0 equiv) was added, and stirring was continued for 15 min. The mixture was then heated to 80 °C under nitrogen protection and stirred for 4 h. The reaction was monitored by TLC (PE:EA = 3:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (PE:EA = 3:1) to give intermediate 23a (2.22 g). Yield: 56%. A pale yellow solid powder.

[0067] Example 25: Synthesis of 1-(7-fluoro-2-(2-hydroxypropyl-2-yl)-6-((trimethylsilyl)ethynyl)quinoline-3-yl)pyridin-1-onium (24a)

[0068] Intermediate 23a (2.22 g, 9.44 mmol, 1.0 equiv) and intermediate 3 (2.45 g, 9.44 mmol, 1.0 equiv) were dissolved in anhydrous ethanol and pre-cooled at 0 °C for 15 min. Then, intermediate 3 (1.0 equiv) and pyridine (1.0 equiv) were added, and stirring was continued for another 15 min. The mixture was then heated to 85 °C under nitrogen protection and stirred for 36 h. The reaction was monitored by TLC (DCM:MeOH = 10:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 10:1) to give intermediate 24a (2.81 g). Yield: 65%. Yellow solid powder.

[0069] Example 26: Synthesis of 2-(3-amino-7-fluoro-6-((trimethylsilyl)ethynyl)quinoline-2-yl)prop-2-ol (25a)

[0070] Intermediate 24a (2.81 g, 6.13 mmol, 1.0 equiv) was dissolved in anhydrous ethanol, and morpholine (2.8 equiv) was added. The mixture was then heated to 85 °C under nitrogen protection and stirred overnight. The reaction was monitored by TLC (PE:EA = 2:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (PE:EA = 2:1) to give intermediate 25a (872 mg). Yield: 45%. Yellow solid powder.

[0071] Example 27: Synthesis of 2-(3-amino-6-ethynyl-7-fluoroquinolin-2-yl)prop-2-ol (26a)

[0072] Intermediate 25a (872 mg, 2.76 mmol, 1.0 equiv) was dissolved in anhydrous methanol, and tetrabutylammonium fluoride (1 M solution in THF, 1.2 equiv) was added. The mixture was stirred at room temperature for 2 h under nitrogen protection. The reaction was monitored by TLC (PE:EA = 2:1). After the starting material reacted completely, saturated ammonium chloride was added to terminate the reaction. The reaction mixture was filtered, concentrated, dissolved in dichloromethane, extracted with saturated brine, dried over anhydrous sodium sulfate, concentrated in the organic phase, and purified by column chromatography (PE:EA = 2:1) to give intermediate 26a (545 mg). Yield: 81%. Yellow oil.

[0073] Example 28: Synthesis of 2-(3-amino-6-[1-(2-(2-aminoethoxy)ethyl]-1H-1,2,3-triazol-4-yl)-7-fluoroquinoline-2-yl)prop-2-ol (27a)

[0074] Intermediate 26a (545 mg, 2.23 mmol, 1.0 equiv) was dissolved in anhydrous tetrahydrofuran, and azido-PEG1-amine (1.1 equiv) and copper(I) 3-methylsalicylate (0.1 equiv) were added. The mixture was stirred at room temperature for 3 h under nitrogen protection. The reaction was monitored by TLC (DCM:MeOH = 5:1). After the starting material was completely reacted, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 5:1) to give intermediate 27a (618 mg). Yield: 74%. Yellow oil.

[0075] Example 29. Synthesis of N1-(2-(2-(4-(3-amino-7-fluoro-2-(2-hydroxypropen-2-yl)quinoline-6-yl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl)-N4-(8-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)octyl)succinamide (28a)

[0076] Linker compound 13 (135 mg, 0.42 mmol, 1.2 equiv) was dissolved in anhydrous dichloromethane, and then... O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethylurea tetrafluoroborate (TBTU, 2 equiv) and Et3N (2 equiv) were stirred to dissolve. After the solution became clear, intermediate 27a (130 mg, 0.35 mmol, 1.0 equiv) was added, and nitrogen was purged. The mixture was stirred at room temperature for 5 h. The reaction was monitored by TLC (DCM: MeOH = 20:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM: MeOH = 20:1) to give the target compound 28a (60 mg). Yield: 25%. The product was a pale yellow solid powder. Results are as follows: Figure 4 As shown, 1 H NMR (600 MHz, DMSO- d 6) δ 8.43 (d, J = 4.0 Hz,1H), 8.31 (d, J = 7.9 Hz, 1H), 7.83 (t, J = 5.5 Hz, 1H), 7.71 (t, J = 5.6 Hz,1H), 7.57 (d, J = 12.4 Hz, 1H), 7.36 (s, 1H), 6.98 (s, 2H), 6.43 – 6.05 (m,1H), 6.00 – 5.53 (m, 2H), 4.63 (t, J = 5.3 Hz, 2H), 3.87 (t, J = 5.3 Hz, 2H), 3.44 (t, J = 5.9 Hz, 2H), 3.36 (t, J = 7.1 Hz, 4H), 3.17 (t, J = 5.8 Hz, 2H), 3.09 (q, J = 7.3 Hz, 4H), 2.99 – 2.96 (m, 2H), 2.35 – 2.17 (m, 6H), 1.62 (s,6H), 1.42 – 1.48 (m, 2H), 1.37 – 1.31 (m, 2H).

[0077] Example 30: Synthesis of N1-(2-(2-(4-(3-amino-7-fluoro-2-(2-hydroxypropen-2-yl)quinoline-6-yl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl)-N4-(2-(2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)ethoxy)ethoxy)ethyl)succinamide (29a)

[0078] Linker compound 14 (135 mg, 0.41 mmol, 1.2 equiv) was dissolved in anhydrous dichloromethane, and then... O -(benzotriazol-1-yl)-N,N,N′,N′-tetramethylurea tetrafluoroborate (TBTU, 2 equiv) and Et3N (2 equiv) were stirred to dissolve. After the solution became clear, intermediate 27a (128 mg, 0.34 mmol, 1.0 equiv) was added, and nitrogen was purged. The mixture was stirred at room temperature for 5 h. The reaction was monitored by TLC (DCM: MeOH = 20:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM: MeOH = 20:1) to give the target compound 29a (70 mg). Yield: 30%. The product was a pale yellow solid powder. Results are as follows: Figure 5 As shown, 1 H NMR (600 MHz, DMSO- d 6) δ 7.60 – 7.56 (m, 1H),7.52 – 7.49 (m, 1H), 7.42 (s, 1H), 7.35 (q, J = 5.5, 4.2 Hz, 1H), 7.26 – 7.21(m, 2H), 7.01 (s, 2H), 6.87 (s, 2H), 5.75 (s, 1H), 4.74 (d, J = 2.7 Hz, 4H), 4.66 – 4.62 (m, 10H), 4.59 (d, J = 10.9 Hz, 2H), 3.09 (q, J = 7.3 Hz, 4H), 2.32 – 2.26 (m, 4H), 1.60 (s, 6H).

[0079] Example 31: Synthesis of 2-amino-4-methoxy-5-((trimethylsilyl)ethynyl)benzaldehyde (23b)

[0080] Intermediate 22 (16 g, 57.77 mmol, 1.0 equiv), triphenylphosphine (0.02 equiv), bis-(triphenylphosphine)-dichloropalladium (0.01 equiv), and CuI (0.03 equiv) were dissolved in a DMF / Et3N solution that had been degassed by bubbling. Nitrogen gas was then introduced, followed by the addition of trimethylsilylacetylene (1.7 equiv). The reaction was stirred at room temperature for 22 h, and the reaction was monitored by TLC (PE:EA = 2:1). After the reactants had completely reacted, the reaction system was concentrated by vacuum distillation. The residue was dissolved in dichloromethane and washed with 1 M hydrochloric acid solution. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (PE:EA = 2:1) to obtain intermediate 23b (11.0 g). Yield: 77%. The product was a pale yellow solid powder.

[0081] Example 32: Synthesis of 1-(2-(2-hydroxypropyl-2-yl)-7-methoxy-6-((trimethylsilyl)ethynyl)quinoline-3-yl)pyridine-1-onium (24b)

[0082] Intermediate 23b (11.0 g, 44.52 mmol, 1.0 equiv) and intermediate 3 (11.53 g, 44.52 mmol, 1.0 equiv) were dissolved in anhydrous ethanol and pre-cooled at 0 °C for 15 min. Then, intermediate 3 (1.0 equiv) and pyridine (1.0 equiv) were added, and stirring was continued for another 15 min. The mixture was then heated to 85 °C under nitrogen protection and stirred for 36 h. The reaction was monitored by TLC (DCM:MeOH = 10:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 10:1) to give intermediate 24b (15.49 g). Yield: 74%. Yellow solid powder.

[0083] Example 33: Synthesis of 2-(3-amino-7-methoxy-6-((trimethylsilyl)ethynyl)quinoline-2-yl)prop-2-ol (25b)

[0084] Intermediate 24b (15.49 g, 32.95 mmol, 1.0 equiv) was dissolved in anhydrous ethanol, and morpholine (2.8 equiv) was added. The mixture was then heated to 85 °C under nitrogen protection and stirred overnight. The reaction was monitored by TLC (PE:EA = 2:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (PE:EA = 2:1) to give intermediate 25b (4.11 g). Yield: 38%. Yellow solid powder.

[0085] Example 34: Synthesis of 2-(3-amino-6-ethynyl-7-methoxyquinoline-2-yl)prop-2-ol (26b)

[0086] Intermediate 25b (4.11 mg, 12.52 mmol, 1.0 equiv) was dissolved in anhydrous methanol, and tetrabutylammonium fluoride (1M solution in THF, 1.2 equiv) was added. The mixture was stirred at room temperature for 2 h under nitrogen protection. The reaction was monitored by TLC (PE:EA = 2:1). After the starting material reacted completely, saturated ammonium chloride was added to terminate the reaction. The reaction mixture was filtered, concentrated, dissolved in dichloromethane, extracted with saturated brine, dried over anhydrous sodium sulfate, concentrated in the organic phase, and purified by column chromatography (PE:EA = 2:1) to give intermediate 26b (2.5 g). Yield: 78%. It was a pale yellow solid powder.

[0087] Example 35: Synthesis of 2-(3-amino-6-[1-(2-(2-aminoethoxy)ethyl]-1H-1,2,3-triazol-4-yl)-7-methoxyquinoline-2-yl)prop-2-ol (27b)

[0088] Intermediate 26b (2.5 g, 9.76 mmol, 1.0 equiv) was dissolved in anhydrous tetrahydrofuran, and azido-PEG1-amine (1.1 equiv) and copper(I) 3-methylsalicylate (0.1 equiv) were added. The mixture was stirred at room temperature for 3 h under nitrogen protection. The reaction was monitored by TLC (DCM:MeOH = 5:1). After the starting material was completely reacted, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 5:1) to give intermediate 27b (2.78 g). Yield: 72%. Pale yellow solid.

[0089] Example 36. Synthesis of N1-(2-(2-(4-(3-amino-2-(2-hydroxypropen-2-yl)-7-methoxyquinoline-6-yl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl)-N4-(8-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)octyl)succinamide (28b)

[0090] Linker compound 13 (136 mg, 0.42 mmol, 1.2 equiv) was dissolved in anhydrous dichloromethane, and then... O -(benzotriazol-1-yl)-N,N,N′,N′-tetramethylurea tetrafluoroborate (TBTU, 2 equiv) and Et3N (2 equiv) were dissolved by stirring. After the solution became clear, intermediate 27b (135 mg, 0.35 mmol, 1.0 equiv) was added, and nitrogen was purged. The mixture was stirred at room temperature for 5 h. The reaction was monitored by TLC (DCM:MeOH = 20:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 20:1) to give the target compound 28b (56 mg). Yield: 23%. The product was a pale yellow solid powder. Results are as follows: Figure 6 As shown, 1 H NMR (600 MHz, DMSO- d 6) δ 8.44 (s, 1H), 8.33 (s,1H), 7.83 (t, J = 5.6 Hz, 1H), 7.71 (t, J = 5.6 Hz, 1H), 7.33 (dd, J = 28.3,22.6 Hz, 2H), 6.99 (s, 2H), 5.75 (s, 2H), 5.69 – 5.34 (m, 1H), 4.61 (t, J =5.3 Hz, 2H), 4.00 (s, 3H), 3.86 (t, J = 5.3 Hz, 2H), 3.44 (t, J = 6.0 Hz,2H), 3.37 (t, J = 7.1 Hz, 2H), 3.19 (q, J = 5.8 Hz, 2H), 2.98 (q, J = 6.7 Hz,2H), 2.35 – 2.21 (m, 6H), 1.63 (s, 6H), 1.49 – 1.42 (m, 2H), 1.37 – 1.29 (m,2H), 1.25 – 1.17 (m, 6H).

[0091] Example 37, N1-(2-(2-(4-(3-amino-7-fluoro-2-(2-hydroxypropen-2-yl)quinoline-6-yl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl)N4-(2-(2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)ethoxy)ethoxy)ethyl)succiamide (29b)

[0092] Linker compound 14 (136 mg, 0.41 mmol, 1.2 equiv) was dissolved in anhydrous dichloromethane. O-(benzotriazol-1-yl)-N,N,N',N'-tetramethylurea tetrafluoroborate (TBTU, 2 equiv) and Et3N (2 equiv) were added, and the mixture was stirred until dissolved. After the solution became clear, intermediate 27b (133 mg, 0.35 mmol, 1.0 equiv) was added, and the mixture was purged with nitrogen and stirred at room temperature for 5 h. The reaction was monitored by TLC (DCM:MeOH = 20:1). After the starting material reacted completely, the reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 20:1) to give target compound 29b (76 mg). Yield: 31%. The product was a pale yellow solid powder. Results are as follows: Figure 7 As shown, 1 H NMR (600 MHz, DMSO- d 6) δ 8.46 – 8.42 (m, 1H), 8.34 (s, 1H), 7.83 (t, J = 5.7 Hz, 1H), 7.79 (t, J = 5.7 Hz, 1H), 7.14 – 7.10(m, 1H), 7.03 –7.08(m, 1H), 7.01 (s, 2H), 5.83 (d, J = 76.2 Hz, 2H), 5.75 (s,1H), 4.61 (t, J = 5.3 Hz, 2H), 4.00 (s, 3H), 3.86 (t, J = 5.4 Hz, 2H), 3.56(t, J = 5.5 Hz, 2H), 3.51 (t, J = 6.0 Hz, 2H), 3.49 – 3.41 (m, 8H), 3.33 (d,J = 6.0 Hz, 2H), 3.19 (d, J = 5.8 Hz, 2H), 3.15 – 3.12 (m, 2H), 1.64 (s, 6H),1.24 (d, J = 5.7 Hz, 2H).

[0093] Experimental Example 1: Compound Stability Experiment After successfully synthesizing target compounds 15, 16, 28a-b, and 29a-b, the stability characteristics of each compound and the positive control were systematically evaluated. First, the stability of each compound and the positive control NS-2 (Reproxalap drug: 2-(3-amino-6-chloroquinoline-2-yl)prop-2-ol) in a physiological saline system was tested. A 5 mM DMSO stock solution of the target compound was prepared. EP tubes were filled with 1800 μL of solvent (physiological saline, sodium hyaluronate eye drops, and rat plasma), followed by 200 μL of DMSO stock solution, yielding 2000 μL of a 500 μM target compound solution. The solution was incubated in a shaker (37 ℃, 200 rpm). At each time point after incubation, 200 μL of the solution was pipetted, 200 μL of mass spectrometry grade methanol was added, and after centrifugation, the supernatant was collected, filtered through a 0.22 μm organic filter, and analyzed by HPLC to calculate the concentration and plot the stability curves of the compounds. All tested compounds showed good stability within 72 h, and their peak areas did not decrease significantly. Figure 7 ).

[0094] The research system was further extended to the artificial tear environment simulated by sodium hyaluronate eye drops (trade name: Waterkeying). Experimental data showed that the stability of the seven tested compounds in this medium was highly consistent with that of the physiological saline system, and all compounds remained stable within 72 hours. Figure 8 ).

[0095] Experimental Example 2: In vitro scavenging experiment of compounds to remove reactive aldehydes To verify the effect of structural modifications to compounds 15 and 16 on their ability to scavenge reactive aldehydes, a parallel comparative experiment was conducted using the positive control drug NS-2. The experiment was based on the detection principle of malondialdehyde (MDA), an end product of lipid peroxidation, and quantitative analysis was performed using the MDA detection kit from Beyotime Biotechnology Co., Ltd. MDA, as an endogenous lipid peroxidation end product, reflects the degree of lipid oxidation under oxidative stress in the body. When cells are subjected to oxidative damage, the oxidative decomposition of polyunsaturated fatty acids generates various metabolites, including MDA. Existing patent data have confirmed that the MDA content in the tears of dry eye patients is significantly higher than that of healthy individuals. Due to the characteristics of reactive aldehyde compounds, this study selected MDA as an evaluation index for reactive aldehyde scavenging ability.

[0096] The structural formula of NS-2:

[0097] The experiment monitored the residual MDA levels at different time points (initial, 30 min, 1 h, 2 h, 4 h, 12 h, 24 h, and 48 h), and found that the clearance efficiency of compounds 15, 16, and the positive control drug NS-2 did not show a significant difference at the 48 h endpoint. Figure 9 This result indicates that the structural modification of the target molecule did not significantly affect its scavenging ability for reactive aldehydes, providing crucial experimental evidence for subsequent drug evaluation. To further investigate the effect of substituent electronic effects on aldehyde scavenging ability, the results showed that the scavenging rate of compound 15 was 1.3 times higher than that of NS-2. Compound 15, with superior activity, and its ortho-substituted derivatives (fluorine-substituted 28a and methoxy-substituted 28b) were selected for comparative analysis. Experimental data showed that the electron-donating methoxy derivative 28b exhibited the best scavenging efficiency, with a scavenging rate 1.5 times higher than NS-2 and a scavenging efficiency 1.2 times higher than that of 15; while the electron-withdrawing fluorine-substituted derivative 28a showed the lowest scavenging activity. Therefore, experimental analysis indicates that electron-donating groups can accelerate the nucleophilic addition reaction with the carbonyl group by enhancing the nucleophilicity of the amino group, while electron-withdrawing groups inhibit the reaction process by reducing the electron density of the amino group.

[0098] Experimental Example 3: In vivo anti-mouse dry eye activity test of the compound Based on the stability and scavenging of active aldehydes, compound 15 was selected for in vivo efficacy validation. A benzalkonium chloride-induced dry eye mouse model was constructed to systematically evaluate the in vivo anti-dry eye efficacy of compound 15. The experiment included gradient dosing groups: low / high dose groups of compound 15 (0.1%, 0.5%), a 0.25% NS-2 positive control group, a Normal group (treated with an equal volume of physiological saline), and a model group (treated with 0.2% benzalkonium chloride). On days 0, 3, and 7 of administration, tear secretion volume was measured, ocular surface integrity was assessed, and a tear fern test was performed. On day 7, corneal histopathological examination (HE staining) and inflammatory factor levels were simultaneously measured to evaluate the compound's anti-dry eye activity.

[0099] 1. Benzalkonium chloride-induced dry eye syndrome in mice experiment 1.1 Experimental animals: 35 C57BL / 6 females (18~20g), purchased from Jiangsu Huachuang Xinno Pharmaceutical Technology Co., Ltd. 1.2 Administration method: eye drops.

[0100] 1.3 Experimental equipment: 1 mL syringe, 20 mL syringe, LYL-S handheld slit lamp microscope, sodium fluorescein ophthalmic test strip (Qianwanli), phenol red cotton thread for tear testing (Tianjin Jingming), electronic digital caliper, pathological microscope slides, 2-10 μL pipettes, cotton, padding cloth, 0.3 mm capillary tube. 1.4 Experimental reagents: isoflurane, physiological saline, PBS buffer (1×, pH = 7.2~7.4), 4% paraformaldehyde fixative, target compound. 1.5 Experimental Methods: C57BL / 6 mice were acclimatized for 3 days (temperature 25 ℃, humidity 40%). The condition of their eyes was observed daily to ensure that the eyes were normal.

[0101] Mice were first anesthetized with isoflurane, and then bilateral modeling was performed in the mice with 0.2% benzalkonium chloride (5 μL / eye / time, twice a day, 2-10 μL pipette, 8:30; 19:30) for 7 consecutive days. After 7 days of modeling, the modeling operation was continued, and bilateral administration was started (5 μL / eye / time, three times a day, 2-10 μL pipette, 10:00; 14:00; 18:00) for 7 consecutive days.

[0102] 1.6 Evaluation Indicators: Ocular surface examination and tear film examination should be performed on mice on days 0, 3, and 7 after drug administration. After day 7, mice should be sacrificed and their eyeballs and corneas removed for inflammatory factor and corneal HE staining.

[0103] 1.6.1 Ocular surface examination Symptoms of dry eye in animals are assessed by examining changes in the morphology of the cornea and conjunctiva using a slit lamp.

[0104] The main observations of a slit lamp include: (1) whether the width of the tear duct has narrowed; (2) the degree of keratinization of the corneal epithelium, and whether there are abnormalities such as blisters, ulcers, or pannus; (3) whether there are debris on the surface of the cornea and the lower fornix; (4) whether there is congestion or papillary hyperplasia of the conjunctiva; (5) whether the conjunctival sac is loose and wrinkled, whether the eyelid margin is congested, or whether there is irregularity, thickening, or eversion; (6) whether there is clear or yellow serous fluid obstruction at the gland opening, or whether the gland duct is blurred.

[0105] Simultaneously, ocular surface examination should be combined with conjunctival fluorescein staining for further examination. The conjunctival fluorescein staining test involves instilling 1 μL of 1% sodium fluorescein into the conjunctival sac and assessing the staining using a slit-lamp microscope. Based on the degree and area of ​​staining, four levels are assigned: no staining (0 points); scattered sparse punctate staining or staining area less than or equal to 1 / 8 of the total area (1 point); relatively dense punctate staining or staining area between 1 / 8 and 1 / 4 of the total area (2 points); patchy staining or staining area between 1 / 4 and 1 / 2 of the total area (3 points); and staining area greater than or equal to 1 / 2 of the total area (4 points). Finally, the scores for each group are statistically analyzed to create a corneal damage severity map.

[0106] 1.6.2 Tear examination (1) Tear secretion test (Schirmer test) This experiment measures the total secretion of basal and reflex tears after conjunctival stimulation, serving as a direct indicator for assessing tear secretion function. The specific procedure is as follows: On days 0, 3, and 7 after drug administration, mice were anesthetized with isoflurane. Using ophthalmic microforceps, a phenol red cotton suture for tear detection was precisely placed into the conjunctival sac at the outer third of the lower eyelid. After holding for 30 seconds, the suture was removed. Because the suture reacts with the tears upon contact, a color change occurs. The tear secretion level can be quantified by measuring the length (in mm) of the reddened area of ​​the suture.

[0107] (2) Tear ferning test (TFT) The tear fernification test effectively assesses changes in tear composition and mucin function by analyzing the morphological characteristics of tear crystals. The experimental procedure is as follows: Mice anesthetized with isoflurane are given 1 μL of physiological saline to the ocular surface. A 0.3 mm capillary glass tube is used to collect tear samples from the caruncle of the lower palpebral conjunctiva using a siphon effect (care should be taken to avoid contact with ocular surface tissue during the procedure). The sample is evenly spread on a pathological microscope slide and dried at 25 ℃ for 10–20 min. The fern-like crystal morphology of the tear is then observed and recorded using an upright fluorescence microscope.

[0108] Crystallization morphology was classified into grades I to IV. Grade I crystallography showed uniform, fern-like branching with relatively narrow spatial intervals, scoring 1 point; Grade II crystallography showed small crystals with fewer branches and increased intervals, scoring 2 points; Grade III crystallography showed a significant reduction in branches, with intervals increasing to approximately the size of the crystals, scoring 3 points; Grade IV showed no fern-like crystallography, but a small number of amorphous crystals were visible, scoring 4 points. Dry eye model animals generally exhibited typical characteristics of Grade III and IV crystallization. By systematically collecting crystallization images from each group and statistically analyzing the scoring data, a tear fern-like degeneration atlas was finally constructed.

[0109] 1.6.3 Corneal HE staining Hematoxylin-Eosin (HE) staining, through nuclear-cytoplasmic contrast, is used to assess corneal histopathological changes, including epithelial integrity, stromal structure, and inflammatory cell infiltration. After the experiment, mice were euthanized by cervical dislocation, and corneal tissue was fixed in 4% paraformaldehyde fixative for 72 h. The tissue was then dehydrated with graded ethanol (70%–100%), cleared with xylene, embedded in paraffin, and prepared into tissue sections approximately 5 μm thick. The sections were cleared and hydrated, stained with HE, mounted, and photographed under an upright fluorescence microscope to observe the histopathological changes in the corneal tissue.

[0110] The characteristics of a normal cornea are: (1) Epithelial layer: composed of 4-5 layers of tightly packed squamous epithelial cells; (2) Matrix layer: parallel collagen fiber bundles, without inflammatory cells; (3) Endothelial layer: a single layer of flat cells continuously covering the cornea. Pathological changes diagnostic criteria: (1) Epithelial damage: reduction or excessive proliferation of basal cells, nuclear pyknosis or cell shedding; (2) Matrix abnormalities: disordered collagen structure, activation of keratinocytes; (3) Inflammatory infiltration: aggregation of neutrophils / lymphocytes; (4) Neovascularization: formation of endothelial cell lumen structures.

[0111] 1.6.4 Detection of corneal and conjunctival inflammatory factors The occurrence of dry eye syndrome is closely related to ocular surface inflammation. Abnormal expression of inflammatory factors (such as IL-1β and TNF-α) is an important biomarker for assessing disease progression and treatment efficacy. The detection of corneal and conjunctival inflammatory factors can be achieved through the following procedure: 1. Sample Collection and Processing: After the experiment, mice were euthanized by cervical dislocation, and the entire eyeball was separated. Then, the eyeball homogenate was lysed, centrifuged, and the protein extract was collected.

[0112] 2. Detection Method: The experiment uses a double-antibody sandwich ELISA method (enzyme-linked immunosorbent assay). The specific procedure is as follows: Preparation before detection: Weigh the mouse eyeballs and add the corresponding tissue homogenate according to the eyeball weight, then homogenize. Remove the kit from the refrigerator 20 minutes in advance to allow it to equilibrate to room temperature. Dilute the 20x concentrated wash buffer with double-distilled water to prepare a 1x working solution. TNF-α standard preparation: Centrifuge at 1000 rpm for 1 minute before opening. Add 0.5 mL of the standard and sample universal diluent, let stand for 15 minutes until dissolved and clear, then gently mix. Prepare a standard curve with the following concentrations: 2000, 1000, 500, 250, 125, 62.5, 31.25, and 0 pg / mL. 0 pg / mL is the blank well. IL-1β standard preparation: Add 1.2 mL of the standard and sample universal diluent again, let stand for 15 minutes until dissolved and clear, then gently mix. Prepare standard curves with the following concentrations: 1000, 500, 250, 125, 62.5, 31.25, 15.6, and 0 pg / mL. The 0 pg / mL solution is the working solution for the biotinylated antibody in the blank wells: 20 minutes before use, dilute the 30x concentrated biotinylated antibody to 1x working solution using biotinylated antibody diluent. For the enzyme conjugate working solution: 20 minutes before use, dilute the 30x concentrated enzyme conjugate to 1x working solution using enzyme conjugate diluent.

[0113] Detection Procedure: Remove the required strips from the sealed bag that has been equilibrated to room temperature. Add standard and sample diluent to the blank wells, and add different concentrations of standard or sample (100 µL / well) to the other wells. Seal the reaction wells with sealing tape and incubate at 37 ℃ in the dark for 90 min. Prepare the biotinylated antibody working solution 20 min in advance. Wash the plate 5 times: Wash manually, shake off the liquid in the wells, and pat dry with clean absorbent paper. Add 350 µL of washing buffer to each well, let stand for 30 seconds, shake off the liquid, and pat dry with thick absorbent paper. Wash a total of 5 times. Add biotinylated antibody diluent to the blank wells, and add biotinylated antibody working solution (100 µL / well) to the other wells. Seal the reaction wells with new sealing tape and incubate at 37 ℃ in the dark for 60 min. Prepare the enzyme conjugate working solution 20 min in advance and store at room temperature (22~25 ℃) in the dark. Wash the plate 5 times. Add enzyme conjugate dilution buffer to blank wells, and add enzyme conjugate working solution (100 µL / well) to the remaining wells. Seal the reaction wells with new sealing tape and incubate at 37 °C in the dark for 30 min. Wash the plate 5 times. Add 100 µL of chromogenic substrate (TMB) to each well and incubate at 37 °C in the dark for 15 min. Add 100 µL of reaction stop solution to each well, mix well, and immediately measure the OD450 value (within 3 min).

[0114] 3. Data Analysis Concentration Quantitative Analysis: Based on the ELISA standard curve, the optical density value (OD) is detected. 450The expression levels of inflammatory factors were converted to absolute concentration values ​​and normalized to per unit weight of tissue (pg / mg).

[0115] 1.7 Statistics All data were statistically analyzed using GraphPad Prism 8 software.

[0116] 2 Results 2.1 Tear secretion volume detection experiment The phenol red cotton thread method was used to quantitatively detect tear secretion, objectively reflecting the level of tear production by measuring changes in the length of the wetted cotton thread. Experimental results are as follows: Figure 10 As shown, the tear secretion in the model group and each drug administration group after modeling showed statistically significant differences compared with the Normal group (p<0.01), confirming the successful establishment of the benzalkonium chloride-induced dry eye model.

[0117] On day 3 after drug intervention, tear secretion recovered in all treatment groups. Notably, the efficacy of compound 15 showed a dose-dependent increase, and tear production in the 5% concentration group was higher than that in the 0.25% NS-2 positive drug group to varying degrees. By day 7 of drug administration, tear secretion in all treatment groups tended to stabilize at a plateau. Quantitative analysis showed that tear secretion in the 0.5% compound 15 group, 0.1% compound 15 group, and 0.25% NS-2 group recovered to levels comparable to the normal group on day 7. This result suggests that high-dose compound 15 not only has the advantage of a rapid onset of action (significant efficacy within 3 days) but also achieves therapeutic effects comparable to physiological states.

[0118] 2.2 Ocular surface examination The degree of damage to the cornea and conjunctiva is assessed using a slit-lamp microscope system to reflect the pathological state of dry eye. Ocular surface examination is performed using white light to obtain the morphological characteristics of the unstained ocular surface. Fluorescein staining assessment involves instilling 1% fluorescein solution (1 μL) and recording the fluorescein-stained area of ​​the cornea using cobalt blue light excitation mode. Damage is scored based on the extent of corneal damage (0-4 points, with higher scores indicating more severe damage).

[0119] The results are as follows Figure 11 As shown, seven days after modeling, the corneal damage scores in the model group and all treatment groups were statistically significantly higher than those in the Normal group, confirming the successful establishment of the dry eye model. On day 3 of drug administration, the corneal repair efficiency in the 0.5% compound 15 group was significantly higher than that in other intervention groups, and superior to the 0.1% compound 15 group and the 0.25% NS-2 positive control group. With continued intervention until day 7, the 0.5% compound 15 group achieved almost complete corneal epithelial repair.

[0120] 2.3 Tear-induced fern experiment The tear fern test was used to quantitatively evaluate the crystallization characteristics of the tear film. Tear samples were collected from the lacrimal caruncle of the lower eyelid conjunctival sac using a capillary tube, and after natural drying, the crystal morphology was observed and a crystallization index (0-4) was assigned using an upright fluorescence microscope (200×). The normal group showed a typical Grade I fern-like structure, characterized by dense, fern-like branches. The model group exhibited Grade III crystallization characteristics, with broken branches, increased lattice spacing, and randomly distributed amorphous crystals.

[0121] Experimental results are as follows Figure 12 As shown, seven days after modeling, the tear crystallization scores of the model group and each treatment group were significantly higher than those of the Normal group, confirming the successful establishment of the dry eye model. On the third day of administration, compound 15 showed a significant improvement in crystal morphology. By the seventh day after intervention, the crystal repair effect of the compound 15 group (0.1% and 0.5%) remained superior to that of the positive control group.

[0122] 2.4 Inflammatory Factor Measurement Experiment In this experiment, mice were sacrificed on day 7 after drug administration and mouse eyeball tissue was collected. Tissue lysate was prepared by low-temperature homogenization, and the concentrations of pro-inflammatory cytokines TNF-α and IL-1β were quantitatively detected by double antibody sandwich ELISA (kit purchased from Xinbosheng).

[0123] Experimental results are as follows Figure 13 As shown, the expression levels of pro-inflammatory cytokines TNF-α and IL-1β in the model group were 2.5-fold and 2.0-fold higher than those in the normal group, respectively (p<0.0001), confirming that dry eye syndrome is accompanied by a significant inflammatory response. The 0.5% compound 15 treatment group showed the strongest anti-inflammatory effect, and its TNF-α and IL-1β levels were not statistically different from those in the normal group (p>0.05).

[0124] The above description is merely an embodiment of the present invention, and the scope of protection of the present invention is not limited to these specific embodiments, but is determined by the claims of the present invention. For those skilled in the art, the present invention can have various modifications and variations. Any modifications, equivalent substitutions, improvements, etc., made within the technical concept and principle of the present invention should be included within the scope of protection of the present invention.

Claims

1. A small molecule active aldehyde scavenger compound, characterized in that, The compounds include compounds or derivatives thereof with the following structures: ； R1 includes hydrogen and C. 1-3 Alkyl, C 2-3 alkenyl, C 2-3 alkynyl group, C 1-3 Alkoxy, C 3-5 cycloalkyl, C 3-5 Cycloalkyloxy, halogen, nitro, cyano, trifluoromethyl, 3-7 membered heterocyclic groups, unsubstituted or Ra-substituted C 6-10 aryl, unsubstituted or Ra-substituted C 6-10 Aryl groups, each Ra may be the same or different, and are independently selected from halogens, C 1-5 Alkyl, C 1-5 Alkoxy, C 6-10 Aryl acyl group. Preferably, R1 is a halogen, C 1-3 At least one of alkoxy, trifluoromethyl, nitro, and cyano groups; Wherein R2 includes at least one of the following structures: 。 2. The compound according to claim 1, characterized in that, The general structural formula of the connector is as follows, where X is selected from carbon atom, nitrogen atom, and oxygen atom, and n is 0~3; 。 3. The compound according to claim 1, characterized in that, The derivatives of the compound include at least one of the following: enantiomers, diastereomers, geometric isomers, free forms, and pharmaceutically acceptable salts, hydrates, solvates, and esters.

4. A composition, characterized in that, The composition comprises any one of the compounds of claims 1-4 and pharmaceutically acceptable additives.

5. The composition according to claim 5, characterized in that, The additives include at least one of the following: binders, fillers, diluents, disintegrants, suspending agents, suspending aids, sustained-release agents, controlled-release agents, lyophilization protectants, coating agents, enteric materials, lubricants, flow aids, anti-adhesion agents, sweeteners, flavoring agents, plasticizers, opacifiers, solubilizers, humectants, solvents, osmotic pressure regulators, colorants, pigments, surfactants, emulsifiers, water-soluble matrices, fat-soluble matrices, oleic matrices, pore-forming agents, gelling agents, preservatives, buffers, chelating agents, and antioxidants.

6. The use of the compound according to any one of claims 1-3 or the composition according to any one of claims 4-5 in any of the following aspects: A1. Application in the preparation of products that promote tear secretion; A2. Application in the preparation of products that promote corneal damage repair; A3. Applications in preparing products with improved crystal morphology; A4. Applications in the preparation of products that inhibit the expression of pro-inflammatory factors; A5. Application in the preparation of products that promote epithelial proliferation and accelerate corneal structural reconstruction.

7. The use of the compound of any one of claims 1-3 or the composition of any one of claims 4-5 in the preparation of a medicament for the treatment and / or prevention of eye diseases, cataracts, keratoconus, Fuchs' endothelial dystrophy in the cornea, uveitis, allergic conjunctivitis, ocular pemphigoid scarring, and refractive keratotomy (PRK) healing disorders.

8. The use of the compound of any one of claims 1-3 or the composition of any one of claims 4-5 in the preparation of a medicament for treating and / or preventing corneal healing-related diseases, diseases related to tear lipid degradation or lacrimal gland dysfunction, or inflammatory eye diseases.

9. The use of the compound of any one of claims 1-3 or the composition of any one of claims 4-5 in the preparation of a medicament for treating and / or preventing eye diseases.

10. The application according to any one of claims 7-9, characterized in that, The drug is in at least one of the following forms: solid, liquid, or gas.

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