A compound and supramolecular nucleoside hydrogel for preventing and / or treating periodontitis and a preparation method and use thereof

Abstract

The application provides a compound and a supramolecular nucleoside hydrogel for preventing and / or treating periodontitis, a preparation method and an application, and belongs to the technical field of biomedical materials.The compound is a compound shown in formula I, a salt or a stereoisomer thereof.The compound shown in formula I can form a stable hydrogel with cyanuric acid in pure water, the hydrogel has good biocompatibility, has no acute toxicity in the body, and improves the problem that the degradation time of 2-FA hydrogel is short; compared with 2-FA hydrogel, the hydrogel has stronger inhibition on osteoclast precursor cells, can more effectively inhibit the differentiation of osteoclasts, and can better reduce the level of alveolar bone absorption, so that the hydrogel can play a more effective effect of preventing and / or treating periodontitis; in addition, the hydrogel is safer to use.Therefore, the hydrogel can be used as a potential drug for preventing and / or treating periodontitis.

Description

Technical Field

[0001] This invention belongs to the field of biomedical materials technology, specifically relating to a compound for the prevention and / or treatment of periodontitis, a supramolecular nucleoside hydrogel, its preparation method, and its uses. Background Technology

[0002] Periodontal infection, leading to periodontal pocket formation, alveolar bone resorption, and tooth loosening, is a major cause of tooth loss in adults. The Fourth National Oral Health Epidemiological Survey showed that the incidence of periodontitis was as high as 87.4%. Prevention and treatment of periodontitis are extremely important for improving oral and overall health in my country. However, the repair effect after alveolar bone loss caused by inflammation is poor, and effectively reducing alveolar bone resorption is a challenging issue in the treatment of periodontitis. Osteoclasts are the only cells responsible for bone resorption and exhibit overactivation during the progression of periodontitis. Early intervention of osteoclast function is a new strategy to reduce alveolar bone loss. Pathogenic bacteria in the periodontal microenvironment are the main initiating factors promoting osteoclast activation. Therefore, a dual treatment strategy of antibacterial action and inhibition of osteoclast differentiation should be constructed to stabilize and eliminate the activation effect of the periodontal microenvironment on osteoclasts, thereby reducing the level of alveolar bone resorption.

[0003] Patent CN114099530B discloses an injectable supramolecular hydrogel self-assembled from 2-amino-2'-fluoro-2'-deoxyadenosine (2-FA) that possesses both antibacterial and osteoclast-inhibiting functional biological characteristics, showing potential application in the treatment of periodontitis. In terms of antibacterial activity, the 2-FA hydrogel exhibits broad-spectrum antibacterial properties, effectively inhibiting the activity of *P. gingivalis*, *Streptococcus mutans*, and *Staphylococcus aureus*. Regarding osteoclast inhibition, the 2-FA hydrogel inhibits the expression of cathepsin K, a marker of osteoclast bone resorption. However, the in vivo degradation time of this 2-FA hydrogel is relatively short (6 hours), while drugs injected into periodontal pockets / gingival sulci require a longer degradation time to match the abnormal bone metabolism cycle in periodontitis. Therefore, further optimization of the 2-FA molecular structure is needed for its application in periodontal treatment. Summary of the Invention

[0004] To address the problems existing in the current treatment of periodontitis with 2-amino-2'-fluoro-2'-deoxyadenosine, this invention provides a compound for the prevention and / or treatment of periodontitis, a supramolecular nucleoside hydrogel, a preparation method, and its uses.

[0005] This invention provides compounds of Formula I, their salts, or stereoisomers thereof:

[0006]

[0007]

[0008] in:

[0009] L is a linking group, selected from phenoxy, triazole, maleimide, and -NH(CH2). a -、-S(CH2) a -、-O(CH2) a -; a is an integer selected from 0 to 8;

[0010] n is an integer selected from 1 to 10000;

[0011] R is selected from hydrogen and C1 to C5 alkyl groups.

[0012] In this invention, the phenoxy structure is as follows:

[0013] Furthermore, the compound is as shown in Formula II or Formula III:

[0014]

[0015] in:

[0016] n is an integer selected from 1 to 10000;

[0017] R is selected from hydrogen and C1 to C5 alkyl groups.

[0018] Furthermore, the compound is one of the following compounds:

[0019]

[0020] In 2-FAPEG1000, the average molecular weight of the PEG fraction is 1000.

[0021] The present invention also provides the use of the aforementioned compounds, their salts or stereoisomers thereof in the preparation of hydrogels for the prevention and / or treatment of periodontitis.

[0022] The present invention also provides a hydrogel for the prevention and / or treatment of periodontitis, which is obtained by dissolving the aforementioned compound, its salt or stereoisomer or cyanuric acid in water.

[0023] Furthermore, the molar ratio of the aforementioned compound, its salt or stereoisomer or cyanuric acid is 1:(1-5);

[0024] Preferably, the molar ratio of the aforementioned compound, its salt or stereoisomer, and cyanuric acid is 1:1.

[0025] Furthermore, the concentration of the aforementioned compound, its salt, or its stereoisomer dissolved in water is 1–15 wt%.

[0026] Preferably, the concentration of the aforementioned compound, its salt, or its stereoisomer dissolved in water is 2.4 wt% to 13.4%.

[0027] The present invention also provides a method for preparing the aforementioned hydrogel, which includes the following steps:

[0028] The aforementioned compound, its salt or stereoisomer, and cyanuric acid were dissolved in water under heating conditions until a clear solution was obtained. After standing and cooling at room temperature, a hydrogel was obtained.

[0029] The present invention also provides the use of the aforementioned compounds, their salts or stereoisomers or the aforementioned hydrogels in the preparation of medicaments for the prevention and / or treatment of periodontitis;

[0030] Preferably, the drug is a drug that reduces alveolar bone loss caused by periodontitis.

[0031] The present invention also provides a medicament for the prevention and / or treatment of periodontitis, which is prepared by adding pharmaceutically acceptable excipients or auxiliary ingredients to the aforementioned compound, its salt or its stereoisomer or the aforementioned hydrogel as the active ingredient.

[0032] This invention optimizes the molecular structure of 2-amino-2'-fluoro-2'-deoxyadenosine (2-FA) nucleoside to prolong the degradation time of hydrogels in vivo. The invention employs a two-step chemical reaction to directly link the 2PEG chain functional group to the 8-position of the 2-FA nucleoside molecule, obtaining a novel gel molecule, 8'-PEG 2-FA (2-FA2PEG). Gel performance screening revealed that 2-FA2PEG exhibits excellent biocompatibility and biosafety, and it can form a stable hydrogel with cyanuric acid in pure water, demonstrating injectability. Subcutaneous injection of the 2-FA2PEG gel resulted in a retention time in vivo between 48 and 60 hours, achieving optimized in vivo degradation time without acute in vivo toxicity.

[0033] The biological functions of the novel 2-FA2PEG hydrogel were evaluated. Comparison of the osteoclast-inhibiting efficiency of 2-FA2PEG with that of 2-FA revealed that 2-FA2PEG had a lower half-maximal inhibitory concentration (WMC) for osteoclast precursor cells, exhibited higher efficiency in inhibiting osteoclast precursor cell proliferation than 2-FA, and showed greater selectivity in its action on osteoclast / osteoblast precursor cells compared to 2-FA. Molecular docking simulations showed that 2-FA2PEG has the potential to inhibit osteoclast differentiation by interfering with RANKL / RANK binding, and the application of 2-FA2PEG hydrogel in a periodontitis animal model significantly reduced alveolar bone resorption. These findings suggest that 2-FA2PEG hydrogel holds promise as a novel strategy for the treatment of periodontitis.

[0034] Based on structural inferences, 2-FA6PEG and 2-FAPEG 1000 It has similar effects to 2-FA2PEG.

[0035] In summary, this invention provides a compound of Formula I that can form a stable hydrogel with cyanuric acid in pure water. This hydrogel exhibits good biocompatibility and no acute toxicity when used in vivo. Furthermore, this hydrogel improves upon the short degradation time of 2-amino-2'-fluoro-2'-deoxyadenosine hydrogel, remaining in vivo for 48–60 hours. Moreover, compared to 2-amino-2'-fluoro-2'-deoxyadenosine hydrogel, the hydrogel of this invention has a stronger inhibitory effect on osteoclast precursor cells, more effectively inhibiting osteoclast differentiation and better reducing alveolar bone resorption, thereby exerting a more effective preventive and / or therapeutic effect on periodontitis. In addition, this hydrogel exhibits a greater concentration difference between inhibited osteoclast precursor cells and osteoblast precursor cells, making it safer to use. Therefore, the hydrogel of this invention can serve as a potential drug for the prevention and / or treatment of periodontitis.

[0036] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions, or alterations can be made without departing from the basic technical concept of the present invention.

[0037] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description

[0038] Figure 1 Three synthetic routes for 2-FAPEG compounds and the structures of the three 2-FAPEG compounds are presented.

[0039] Figure 2 The following are the proton, carbon, high-resolution mass spectra and single-crystal structures of the 2-FA2PEG compound: A shows the proton, carbon, and high-resolution mass spectra of the 2-FA2PEG compound; B shows the single-crystal structure of the 2-FA2PEG compound.

[0040] Figure 3 2-FA2PEG hydrogel, 2-FA6PEG hydrogel and 2-FAPEG 1000 An inverted image of a hydrogel vial.

[0041] Figure 4 The results of the evaluation of the biological performance of 2-FA2PEG compound in regulating osteoclast precursor cells and osteoblast precursor cells.

[0042] Figure 5The results of the performance study of 2-FA2PEG hydrogel are as follows: A shows the rheological test results and viscosity variation with shear force after gelation of 2-FA2PEG hydrogel; B shows the in vivo degradation performance of 2-FA2PEG hydrogel; C shows the biocompatibility of 2-FA2PEG hydrogel.

[0043] Figure 6 These are simulated binding sites for 2-FA and 2-FA2PEG compounds with RANKL (PDB entry: 3me2).

[0044] Figure 7 The experimental flowchart and Micro-CT evaluation results of applying 2-FA2PEG hydrogel to treat periodontitis in mice are shown in Figure 1: A is the experimental flowchart; B is the Micro-CT evaluation results. Detailed Implementation

[0045] The raw materials and equipment used in the specific embodiments of the present invention are all known products, obtained by purchasing commercially available products.

[0046] Example 1: Synthesis of 2-FAPEG compound

[0047] The synthetic route of 2-FAPEG compounds is as follows: Figure 1 As shown, the specific synthesis method is as follows:

[0048] (1) Synthesis of 2-FABr: 500 mg of 2-FA (1.74 mmol, 1.0 eq.) was dissolved in 20 mL of MeCN and 5 mL of water in a 100 mL round-bottom flask. 464.5 mg (2.61 mmol, 1.5 eq.) of N-bromosuccinimide was added in portions. The suspension was stirred at room temperature for 60 minutes until 2-FA was completely consumed as monitored by TLC. The crude product was purified by column chromatography (DCM / MeOH, volume ratio 95:5) to obtain 365 mg (1.01 mmol, 58%) of white solid (2-FABr). HRMS(ESI+)[M+H]+calcd m / z for [C10H13BrFN6O3] + :363.0217,found:363.0214. 1H NMR(400MHz, DMSO-d6)δ6.99(s,2H),5.99–5.88(m,2H),5.88–5.69(m,1H),5.62(d,J=6.4Hz,1H),5.20(dd,J=6.9,4.9Hz,1H),4 .58(dq,J=14.7,5.9Hz,1H), 3.93(td,J=6.1,2.9Hz,1H), 3.70(ddd,J=12.2,5.0,3.1Hz,1H), 3.52(ddd,J=12.3,7.0,5.5Hz,1H).

[0049] (2) Synthesis of 2-FA2PEG: 2-FABr (1.34 g, 3.67 mmol, 1.0 eq.), (4-(2-(2-methoxyethoxy)ethoxy)phenyl)boronic acid (1.0 g, 4.04 mmol, 1.1 eq.), tetrakis(triphenylphosphine)palladium Pd(PPh3)4 (229 mg, 0.198 mmol, 0.05 eq.), and anhydrous potassium carbonate (2.12 g, 11.88 mmol, 3 eq.) were added to 50 mL of a 1,4-dioxane:water (1:1, v:v) mixed solution. The reaction was carried out at 100 °C for 6 hours under nitrogen protection. After the conversion of the 2-FABr starting material was completed by TLC, the reaction was quenched by adding water. The mixture was extracted with EA, and the organic phases were combined. The mixture was purified by column chromatography (DCM / MeOH volume ratio 95:5) to obtain 1.16 g (2.42 mmol, 66%) of white solid (2-FA2PEG). HRMS(ESI+)[M+H]+calcd m / z for[C21H28FN6O6] + :479.2054,found:479.2050. 1 H NMR(400MHz,DMSO-d6)δ7.61–7.53(m,2H),7.17–7.09(m,2H),6.99–6.79(m,2H), 5.92–5.83(m,1H),5.83–5.71(m,3H),5.55(s,1H),5.42(dd,J=7.4,4.6Hz,1H),4. 61–4.46(m,1H),4.21–4.14(m,2H),3.94–3.87(m,1H),3.80–3.75(m,2H),3.75–3 .68(m,1H),3.64–3.60(m,1H),3.60–3.51(m,2H),3.50–3.43(m,2H),3.25(s,3H). 13C NMR(101MHz,DMSO-d6)δ159.78,159.50,156.22,152.08,146.73,130.56,121.99,114.74,1 13.04,92.28,90.43,86.88,84.84,71.30,69.73,69.13,68.98,68.85,67.34,61.76,58.09. 19 F NMR(376MHz,DMSO-d6)δ-203.77.

[0050] 2-FA2PEG was dissolved in methanol and recrystallized to obtain colorless, blocky crystals. The crystal parameters were: C 21 H 27 FN6O6; Mr=478.48g·mol-1; monoclinic, space group C 2; α=90°, β=98.280(8)°, γ=90°; Z=4; calc density=1.290g·cm-3; F(000)=1008.0; T=290K; μ=0.101mm-1; miller index ranges, h=31, k=12, l=14; F2=1.054; R1=0.0383, wR2=0.1085.

[0051] (3) Synthesis of 2-FA6PEG: Using 2-FABr (500 mg, 1.38 mmol, 1.0 eq) and 4-hydroxyphenylboronic acid (210 mg, 1.52 mmol, 1.1 eq) as raw materials, 428 mg (1.15 mmol, 83%) of 4-hydroxyphenyl-substituted white solid 2-FA6PEG was obtained by the same method as in step (2). HRMS(ESI+)[M+H]+calcd m / z for[C16H18FN6O4] + :377.1374,found:377.1371. 1H NMR(400MHz,DMSO-d6)δ9.98(s,1H),7.51–7.38(m,2H),6.96–6.90(m,2H),6.89(s,1H),5.86(d,J=5.4Hz,1H),5.82–5.69(m,3H) ,5.56(d,J=6.2Hz,1H),5.45(dd,J=7.4,4.6Hz,1H),4.61–4.46(m,1H),3.96–3.85(m,1H),3.77–3.67(m,1H),3.61–3.48(m,1H). 13 C NMR(101MHz,DMSO-d6)δ160.14,159.21,156.60,152.50,147.68,131.11,120 .74,116.02,113.42,92.72,90.87,87.32,86.99,85.30,69.60,69.45,62.25.

[0052] 2-FAPh (214.5 mg, 0.57 mmol), 17-hydroxy-3,6,9,12,15-pentaheptadecyl-4-methylbenzenesulfonate (250 mg, 0.57 mmol), and anhydrous potassium carbonate (86.7 mg, 0.63 mmol) were weighed into a 25 mL Schlenk tube. 10 mL of methanol was added, and the mixture was heated to 70 °C for 16 hours. The reaction was quenched with water, extracted with EA, and the organic phases were combined. The mixture was purified by column chromatography (DCM / MeOH, v / v 10:1) to obtain 245 mg (0.38 mmol, 67%) of a white solid (2-FA6PEG). HRMS(ESI+)[M+H]+calcd m / z for [C28H42FN6O10] + :641.2946,found:641.2997. 1 H NMR(400MHz,DMSO-d6)δ7.62–7.54(m,2H),7.53–7.46(m,1H),7.18–7.10(m,3H) ,6.93(s,2H),5.92–5.83(m,1H),5.83–5.72(m,3H),5.57(d,J=6.2Hz,1H),5.45( dd,J=7.4,4.7Hz,1H),4.61(d,J=5.5Hz,1H),4.59–4.50(m,1H),4.22–4.15(m,2 H),3.94–3.87(m,1H),3.82–3.76(m,2H),3.76–3.69(m,1H),3.64–3.39(m,19H). 13C NMR(101MHz,DMSO-d6)δ159.78,159.52,156.22,152.09,146.75,145.60,137.73,130.57,128.09,125.53,1 21.98,114.75,113.04,92.30,84.84,72.35,69.94,69.82,69.80,69.78,68.98,68.87,67.35,61.77,60.22.

[0053] (4)2-FAPEG 1000 Synthesis:

[0054] Weigh 2-FABr (2.0 g, 5.5 mmol), triisopropylsilyne (2.47 mL, 11.0 mmol), cuprous iodide (210 mg, 1.1 mmol), Pd(PPh3)4 (636 mg, 0.55 mmol), and triethylamine (2.3 mL, 16.5 mmol) into a 100 mL double-necked flask containing 25 mL of dry DMF. Stir at room temperature for 10 hours, then quench the reaction with water. Extract with EA, and combine the organic phases and evaporate to dryness. Add 20 mL of THF and 2 g of tetrabutylammonium fluoride directly to the evaporated flask, stir at room temperature for 1 hour to remove the silicon protecting group, quench the reaction with water, extract with EA, combine the organic phases, and purify by column chromatography (EA / MeOH, v / v 100:5) to obtain 490 mg (1.59 mmol, 29%) of pale yellow solid (2-FACCH). HRMS(ESI+)[M+H]+calcd m / z for[C12H14FN6O3] + :309.11,found:309.10.

[0055] Add 2-FACCH (50 mg, 0.162 mmol), N3-PEG1000 (179 mg, 0.17 mmol), 0.1 M Cu(OAc)2 aqueous solution (162 μL, 0.0162 mmol), 0.2 M sodium ascorbate aqueous solution (162 μL, 0.0324 mmol), and 2 mL DMSO to a 10 mL Schlenk tube. Incubate at room temperature for 11 hours. After evaporating the solvent, purify directly by column chromatography (DCM / MeOH volume ratio 10:1) to obtain 130 mg (0.098 mmol, 61%) of a pale yellow viscous liquid (2-FAPEG1000). HRMS(ESI+)[M+H]+calcd m / z for [C56H103FN9O25] + :1320.70,found:1320.62.

[0056] Figure 2 A shows the proton, carbon, and high-resolution mass spectra of 2-FA2PEG; Figure 2 B represents the single-crystal structure and single-crystal network of the 2-FA2PEG compound.

[0057] Example 2: Preparation of 2-FAPEG hydrogel

[0058] Preparation of 2-FA2PEG hydrogel: 2-FA2PEG (4.78 mg, 0.05 M, 2.4 wt%) and cyanuric acid (1.29 mg, 0.05 M, 0.65 wt%) were accurately weighed in an equimolar ratio using an electronic scale, and 200 μL of ultrapure water was added. The solution was heated at 100 °C and sonicated until it was fully dissolved and became a clear solution. The solution was allowed to cool at room temperature, and the formation of the gel was further verified by inverting a vial. A milky white gel was observed to form after 15 min.

[0059] Preparation of 2-FA6PEG hydrogel: 2-FA6PEG (12.81 mg, 0.1 M, 6.4 wt%) and cyanuric acid (2.58 mg, 0.1 M, 1.3 wt%) were accurately weighed in an equimolar ratio using an electronic balance. 200 μL of ultrapure water was added, and the solution was heated at 100 °C and sonicated until it was fully dissolved and became a clear solution. The solution was allowed to cool at room temperature, and the formation of the gel was further verified by an inverted vial experiment. A transparent gel was observed to form after 5 min.

[0060] 2-FAPEG 1000 Preparation of hydrogels: Equimolar amounts of 2-FAPEG were accurately weighed using an electronic balance. 1000 (26.71 mg, 0.1 M, 13.4 wt%) and cyanuric acid (2.58 mg, 0.1 M, 1.3 wt%) were added to 200 μL of ultrapure water, heated at 100 °C and sonicated to fully dissolve until a clear solution was formed. The solution was allowed to stand and cool at room temperature, and the formation of the gel was further verified by inverting a vial. A yellow gel was observed to form after 10 min.

[0061] 2-FA2PEG hydrogel, 2-FA6PEG hydrogel and 2-FAPEG 1000 Image of the hydrogel vial upside down Figure 3 As shown: This illustrates the use of 2-FA2PEG, 2-FA6PEG, and 2-FAPEG. 1000 Hydrogels were successfully prepared in both cases.

[0062] The following specific experimental examples demonstrate the beneficial effects of the present invention.

[0063] Experiment 1: Test of the inhibitory effect of 2-FA2PEG compound on osteoclast precursor cell activity

[0064] 1. Experimental Methods

[0065] In this experiment, RAW264.7 and MC3T3-E1 cell lines were used as osteoclast precursor cells and osteoblast precursor cells, respectively. Cells were cultured in 96-well plates at a density of 102. 4 / well. Add 10 μL of 2-FA or 2-FA2PEG solution to 90 μL of cell suspension to make the final drug concentrations 0 mM, 0.05 mM, 0.1 mM, 0.2 mM, 0.5 mM, 1 mM, 2 mM, 5 mM and 10 mM. Incubate at 37 °C for 24 h, and detect the concentrations using the CCK-8 method and convert them to IC50 values.

[0066] 2. Experimental Results

[0067] Cell viability test results as follows Figure 4 As shown:

[0068] The 2-FA2PEG compound had an IC50 of 1.3 mM for inhibiting osteoclast precursor cell activity and 7.5 mM for inhibiting osteoblast precursor cell activity. Compared to 2-FA (IC50 = 8.9 mM), the 2-FA2PEG compound showed a lower IC50 for osteoclast precursor cells and a higher efficiency in inhibiting their proliferation. Meanwhile, the 2-FA2PEG compound exhibits a larger gradient in its half-maximal inhibitory concentration (IC50) for osteoclast precursor cells (RAW264.7) / osteoblast precursor cells (MC3T3-1) (IC50 for osteoclast precursor cells / IC50 for osteoblast precursor cells = 0.179), compared to 2-FA (IC50 for osteoclast precursor cells = 8.9 mM, IC50 for osteoblast precursor cells = 10.9 mM, IC50 for osteoclast precursor cells / IC50 for osteoblast precursor cells = 0.822). Therefore, 2-FA2PEG demonstrates a greater selectivity for osteoclast / osteoblast precursor cell proliferation. Thus, in the bone homeostasis balance between osteoblast osteogenic and osteoclast bone resorption, 2-FA2PEG exhibits a more significant osteogenic potential than 2-FA.

[0069] Experimental Example 2: In vivo application test of 2-FA2PEG hydrogel

[0070] 1. Experimental Methods

[0071] The hydrogel preparation method in this experiment was the same as in Example 2 (2-FA2PEG hydrogel). Rheological tests were performed on the 2-FA2PEG hydrogel; and in vivo degradation and acute toxicity assessments were conducted. The experimental animals were BALB / c mice (weight 20±5g; male; 8 weeks). 2-FA2PEG hydrogel (100 μL / mouse) was subcutaneously injected into the mice. Samples were collected at 0, 12, 24, 36, 48, 60, and 72 hours post-injection to study the in vivo degradation performance of the hydrogel. 2-FA hydrogel was used as a control group (100 μL / mouse). Simultaneously, the condition of various organs was studied 48 hours after 2-FA2PEG hydrogel injection, with the same volume of ultrapure water used as a control group.

[0072] The preparation method of 2-FA hydrogel is as follows: 2-FA is dissolved in ultrapure water (concentration of 2.5wt%), heated to 100℃ and ultrasonically vibrated until it becomes a clear solution. After standing and cooling at room temperature, a milky white gel is formed after 18 minutes.

[0073] 2. Experimental Results

[0074] Gel performance verification: 2.4wt% 2-FA2PEG can form a stable hydrogel with cyanuric acid in pure water, and this hydrogel exhibits shear-thinning properties, that is, the viscosity of the hydrogel decreases with increasing shear rate, indicating that the hydrogel is injectable. Figure 5 A).

[0075] In vivo degradation experiment: Subcutaneous injection of 2.4 wt% 2-FA2PEG hydrogel ( Figure 5 B), whose degradation time is greater than 48h, indicates that its degradation performance is better than that of 2.5wt% 2-FA hydrogel (6h). The in vivo degradation results of 2-FA are from the published paper Advanced Materials (2022:2108300).

[0076] In vivo acute toxicity assessment: 48 hours after injection, no significant damage was observed in the heart, liver, spleen, lungs, or kidneys of the treatment group (2-FA2PEG hydrogel group), indicating that the 2-FA2PEG hydrogel has good biocompatibility. Figure 5 C).

[0077] Experimental Example 3: The inhibitory potential of 2-FA2PEG compound in inhibiting osteoclast differentiation

[0078] 1. Experimental Methods

[0079] This experimental example was obtained from the protein database PDB (… https: / / www.pdbus.orgDownload the X-ray diffraction crystal structure of the mouse RANKL conformation (PDBentry:3me2). Preprocess 2-FA, 2-FA2PEG, and RANKL molecules in Autodock 4.0 and Chem3D 14.0. Molecular docking was performed using 2-FA and 2-FA2PEG as ligands and RANKL as the acceptor; the automatic docking genetic algorithm was run 10 times. The lowest binding energy conformation was selected, and molecular stacking and visualization were performed in Pymol 2.5.

[0080] 2. Experimental Results

[0081] Molecular docking simulations were performed using Autodock software, and the results showed ( Figure 6 ), 2-FA and 2-FA2PEG are located precisely in the RANKL-RANK binding region, and both molecules bind to one of the key RANKL-RANK binding sites, Y. 234 The presence of an interaction suggests that this type of nucleoside molecule may exert its potential to inhibit osteoclast differentiation by binding to the active pocket of RANKL and interfering with the binding of RANKL to RANK.

[0082] Experiment 4: Test of 2-FA2PEG hydrogel reducing alveolar bone loss in a periodontitis model

[0083] 1. Experimental Methods

[0084] In this experiment, C57 / BL6 mice (weight 20±5g; male; 8 weeks) were used to establish a periodontitis model. The experimental method of ligation + bacterial smear was employed (schematic diagram shown in 7A). The specific modeling procedure was as follows: a 0.15mm stainless steel wire was used to ligate the first molar; a suspension of *P. gingivalis* (ATCC 33277) bacteria (density 5x10⁻¹) was prepared using 4% sodium carboxymethyl cellulose. 9 A periodontitis model was established by applying bacteria (CFU / mL, dose 20 μL / mouse) to the gingival sulcus of the maxillary first and second molars every other day for a total of 15 applications. The group without any treatment after establishing the periodontitis model was designated as the periodontitis group (CP group); normal mice served as the control group (Ctrl); and the group treated with 2-FA2PEG hydrogel was designated as the periodontitis + 2-FA2PEG hydrogel group (CP + 2-FAPEG).

[0085] The 2-FA2PEG hydrogel is a 2.4 wt% 2-FA2PEG hydrogel prepared by the method described in Example 2.

[0086] CP+2-FAPEG treatment: On the day of bacterial smear application, 2.4wt% 2-FA2PEG hydrogel was injected into the gingival sulcus of the maxillary first and second molars using a microinjector at a dose of 20 μL / site, once every two days until day 29 of modeling. Mice were sacrificed on day 30 of modeling for alveolar bone resorption analysis (n=10).

[0087] 2. Experimental Results

[0088] 2-FA2PEG hydrogel significantly reduced the cementoenamel-alveolar ridge (CEJ-AB) distance and alveolar bone loss level in periodontitis mice. Figure 7 B). This invention demonstrates that the 2-FA2PEG hydrogel can be used to improve alveolar bone loss and further play a role in the prevention and / or treatment of periodontitis.

[0089] In summary, this invention provides a compound of Formula I that can form a stable hydrogel with cyanuric acid in pure water. This hydrogel exhibits good biocompatibility and no acute toxicity when used in vivo. Furthermore, this hydrogel improves upon the short degradation time of 2-amino-2'-fluoro-2'-deoxyadenosine hydrogel, remaining in vivo for 48–60 hours. Moreover, compared to 2-amino-2'-fluoro-2'-deoxyadenosine hydrogel, the hydrogel of this invention has a stronger inhibitory effect on osteoclast precursor cells, more effectively inhibiting osteoclast differentiation and better reducing alveolar bone resorption, thereby exerting a more effective preventive and / or therapeutic effect on periodontitis. In addition, this hydrogel exhibits a greater concentration difference between inhibited osteoclast precursor cells and osteoblast precursor cells, making it safer to use. Therefore, the hydrogel of this invention can serve as a potential drug for the prevention and / or treatment of periodontitis.

Claims

1. The compound shown in Formula I, its salt, or its stereoisomer: Formula I in: L is a linking group, selected from phenoxy, triazole, maleimide, and -NH(CH2). a -、-S(CH2) a -、-O(CH2) a -; a is an integer selected from 0 to 8; n is an integer selected from 1 to 10000; R is selected from hydrogen and C1~C5 alkyl groups.

2. The compound, its salt, or its stereoisomer according to claim 1, characterized in that: The compound is shown in Formula II or Formula III: Formula II Formula III in: n is an integer selected from 1 to 10000; R is selected from hydrogen and C1~C5 alkyl groups.

3. The compound, its salt, or its stereoisomer according to claim 1, characterized in that: The compound is one of the following compounds: In 2-FAPEG1000, the average molecular weight of the PEG fraction is 1000.

4. Use of the compound, its salt or stereoisomer as claimed in any one of claims 1 to 3 in the preparation of hydrogels for the prevention and / or treatment of periodontitis.

5. A hydrogel for the prevention and / or treatment of periodontitis, characterized in that: It is obtained by dissolving the compound, its salt or stereoisomer, or cyanuric acid in water according to any one of claims 1 to 3.

6. The hydrogel according to claim 5, characterized in that: The molar ratio of the compound, its salt or stereoisomer or the cyanuric acid described in any one of claims 1 to 3 to cyanuric acid is 1:(1 to 5).

7. The hydrogel according to claim 6, characterized in that: The molar ratio of the compound, its salt or stereoisomer or the cyanuric acid described in any one of claims 1 to 3 is 1:

1.

8. The hydrogel according to any one of claims 5 to 7, characterized in that: The concentration of the compound, its salt or stereoisomer as described in any one of claims 1 to 3, when dissolved in water is 1 to 15 wt%.

9. The hydrogel according to claim 8, characterized in that: The concentration of the compound, its salt or stereoisomer as described in any one of claims 1 to 3, when dissolved in water is 2.4 to 13.4 wt%.

10. A method for preparing the hydrogel according to any one of claims 5 to 9, characterized in that: It includes the following steps: The compound, its salt or stereoisomer, and cyanuric acid of any one of claims 1 to 3 are dissolved in water under heating conditions to obtain a clear solution, and then allowed to stand and cool at room temperature to obtain a hydrogel.

11. Use of the compound, its salt or stereoisomer, or the hydrogel according to any one of claims 1 to 3 in the preparation of a medicament for the prevention and / or treatment of periodontitis.

12. The use according to claim 11, characterized in that: The drug mentioned is a drug that reduces alveolar bone loss caused by periodontitis.

13. A medicament for the prevention and / or treatment of periodontitis, characterized in that: It is prepared by adding pharmaceutically acceptable excipients or auxiliary ingredients to the compound, salt or stereoisomer of any one of claims 1 to 3 or the hydrogel of any one of claims 5 to 9 as the active ingredient.

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