Application of tetrahedral framework nucleic acid-resveratrol compound in preparation of medicine for treating retinal neovascular diseases

Abstract

The invention discloses application of a tetrahedral framework nucleic acid-resveratrol compound in preparation of a medicine for treating retinal neovascular diseases. According to the invention, resveratrol molecules are coupled to a DNA single-stranded skeleton of tFNAs through an electrostatic adsorption method, so that the tetrahedral framework nucleic acid-resveratrol compound (tFNAs-RSV) is formed. Researches show that when the tFNAs-RSV compound is used for treating retinal neovascular diseases, the four targets of resisting retinal pathological neovascularization, improving retinal perfusion, promoting retinal physiological vascularization and protecting a retinal nerve vascular unit (RNVU) which are difficult to achieve simultaneously by single medication of tFNAs or RSV are achieved, and a synergistic interaction effect is exerted; and a breakthrough solution is provided for precise treatment of the retina neovascular diseases.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and more specifically, to the use of a tetrahedral framework nucleic acid-resveratrol complex in preparing a drug for treating retinal neovascular diseases. Background Art

[0002] Retinal neovascular diseases (RNDs) primarily result from a disruption in the dynamic balance between pro-angiogenic and anti-angiogenic factors within the retina / choroid, triggering abnormal vascular proliferation and severely impairing visual function, impacting the quality of life for millions of patients worldwide. Clinical manifestations encompass a wide range of diseases, including retinopathy of prematurity (ROP), a common condition in infants and young children, proliferative diabetic retinopathy (PDR), and familial exudative vitreoretinopathy (FEVR), as well as retinal vein occlusion (RVO), a common condition in young and middle-aged adults, and wet age-related macular degeneration (wAMD), a common condition in the elderly.

[0003] Neurons, glial cells, and vascular cells within localized regions of retinal tissue are functionally coupled and interdependent. The retinal neurovascular unit (RNVU) comprises glial cells (Müller cells, astrocytes, and microglia), vascular cells (endothelial cells and pericytes), and neurons (ganglion cells and bipolar cells). Located in the inner retina, the RNVU's components are closely interconnected. While maintaining the integrity of the inner blood retinal barrier (IBRB), the RNVU dynamically regulates blood flow to meet the metabolic needs of neurons. Changes in photoreceptor cells in the outer retina can also affect RNVU homeostasis. Retinal neovascular diseases are often accompanied by retinal neuronal degeneration and impaired RNVU function.

[0004] Intravitreal injection of anti-vascular endothelial growth factor (VEGF) drugs is currently the first-line treatment for retinal neovascularization, with promising results. However, this therapy has certain limitations. It only reduces pathological retinal neovascularization without protecting damaged retinal vessels and structural neurons. Furthermore, it requires multiple injections and follow-up visits, placing a significant burden on patients. Therefore, there is an urgent need to develop therapeutic agents that can inhibit retinal neovascularization while improving the restoration of physiological retinal vascular structure and protecting the function of the retinal neurovascular unit.

[0005] Resveratrol (3,5,4'-trihydroxystilbene, RSV) is a non-flavonoid polyphenolic compound with a wide range of biological activities, including but not limited to antioxidant, anti-inflammatory, anti-diabetic, anti-obesity, anti-hypertensive, and anti-tumor activities. Chen Lei et al. reported that resveratrol upregulates SIRT1 and p-FOXO3a expression, suggesting that resveratrol may alleviate oxidative stress, inflammation, and neovascularization in the cornea after alkali burns through the SIRT1 / FOXO3a pathway. However, the cornea and retina differ significantly in location, structure, function, and blood supply, leading to significant differences in the causes, hazards, treatments, and therapeutic objectives of their neovascularization. The cornea, located at the anterior pole of the eye, is highly transparent and sensitive, primarily responsible for refraction. The cornea itself is avascular, and corneal neovascularization is caused by abnormal invasion of the limbal vascular network into the cornea, such as in chemical burns. Because the cornea serves as the "refractive window," treatment primarily focuses on anti-neovascular therapy to minimize the effects on its transparency. The retina, located in the inner layer of the eyeball (fundus), adheres to the inner surface of the choroid and contains a photoreceptor layer of neural tissue primarily responsible for light perception and signal conversion. The retina itself contains normal blood vessels and optic nerves. Retinal neovascularization is abnormal blood vessels induced by retinal ischemia and hypoxia. Because the retina is the "photosensitive center," its neovascularization is more serious (higher risk of blindness) due to its tendency to bleed and damage photoreceptor cells. Therefore, as mentioned above, in addition to anti-angiogenic treatment, it is also necessary to restore the retinal physiological vascular structure and protect the function of the retinal neurovascular unit.

[0006] Tetrahedral framework nucleic acids (tFNAs) are a new type of bionanomaterial. tFNAs are tetrahedral structures formed by denaturation and renaturation of four single-stranded DNA strands, followed by complementary base pairing between the strands. They are easy to synthesize, structurally stable, and highly biocompatible, with excellent mechanical properties and promising potential in the biomedical field. Patent CN112843081A discloses a DNA tetrahedron-resveratrol complex, a mixture of DNA tetrahedron-resveratrol and resveratrol. Intraperitoneal injection of this DNA tetrahedron-resveratrol complex improves glucose and insulin tolerance in mice fed a high-fat, high-sugar diet, effectively treating obesity and improving or even curing obesity-induced insulin resistance. tFNAs can also be used to treat metabolic syndrome, type 2 diabetes, hypertension, hyperuricemia, dyslipidemia, cardiovascular and cerebrovascular diseases, polycystic ovary syndrome, Alzheimer's disease, and cancers caused by insulin resistance. Another patent, CN116747237A, discloses the use of a resveratrol-tetrahedral framework nucleic acid complex in the preparation of a drug for preventing and / or treating diabetes or its complications. Administration of the complex via tail vein injection in db / db mice with peripheral neuropathy demonstrated significant therapeutic effects on diabetic peripheral neuropathy. Furthermore, the complex increased sensitivity to mechanical, cold, and heat stimuli, alleviated neurovascular disease, alleviated sciatic nerve demyelination, and rescued sciatic nerve apoptosis. However, the use of a tetrahedral framework nucleic acid-resveratrol complex for retinal neovascularization has not yet been reported. Summary of the Invention

[0007] The purpose of the present invention is to overcome the above-mentioned defects and deficiencies in the prior art and to provide a tetrahedral framework nucleic acid-resveratrol complex for use in the preparation of a drug for treating retinal neovascular diseases.

[0008] The above-mentioned object of the present invention is achieved through the following technical solutions: The present invention couples a specific number of resveratrol molecules to the single-stranded DNA backbone of tFNAs via electrostatic adsorption, enabling the resulting tFNAs to stably load RSV molecules, resulting in a tetrahedral framework nucleic acid-resveratrol complex (tFNAs-RSV). Furthermore, by establishing a mouse model of oxygen-induced retinopathy, the therapeutic efficacy of tFNAs-RSV on retinal neovascular disease was investigated. Results demonstrated that the tFNAs-RSV complex exhibited a four-fold synergistic effect: inhibiting retinal neovascularization, reducing retinal nonperfused areas, promoting physiological retinal vascular remodeling, and enhancing the formation of the retinal neurovascular unit (RNVU). The efficacy was significantly superior to that of RSV or tFNAs alone, demonstrating a synergistic effect and effective treatment of retinal neovascular disease.

[0009] Therefore, the present invention provides the use of a tetrahedral framework nucleic acid-resveratrol complex in the preparation of a drug for treating retinal neovascular diseases, wherein the tetrahedral framework nucleic acid-resveratrol complex is a complex formed by mixing a tetrahedral framework nucleic acid and resveratrol; the molar ratio of the tetrahedral framework nucleic acid to resveratrol is 1:(300-350), that is, the molar ratio of the two materials is 1:(300-350).

[0010] Furthermore, the tetrahedral framework nucleic acid is formed by four single-stranded DNAs through base complementary pairing, and the nucleotide sequences of the four single-stranded DNAs are shown as SEQ ID No. 1 to 4 respectively.

[0011] Furthermore, the preparation method of the tetrahedral framework nucleic acid is to maintain four single-stranded DNAs in a buffer at 85-105° C. for 5-15 minutes, and then maintain it at 2-8° C. for 10-30 minutes.

[0012] Preferably, the tetrahedral framework nucleic acid is prepared by maintaining four single-stranded DNAs in a TM buffer having a pH of 8.0 at 95° C. for 10 min, and then maintaining at 4° C. for 20 min.

[0013] Specifically, the drug achieves treatment by resisting retinal pathological neovascularization, improving retinal perfusion (reducing the area of ​​retinal non-perfusion area), promoting retinal physiological vascular reconstruction, and protecting retinal neurovascular unit. That is, the present invention provides the use of tetrahedral framework nucleic acid-resveratrol complex in the preparation of drugs for resisting retinal pathological neovascularization, improving retinal perfusion (reducing the area of ​​retinal non-perfusion area), promoting retinal physiological vascular reconstruction and protecting retinal neurovascular unit.

[0014] Furthermore, retinal neovascular diseases include, but are not limited to, one or more of diabetic retinopathy, retinopathy of prematurity, retinal vein occlusion, retinal periphlebitis, age-related macular degeneration, polypoidal choroidal vasculopathy, choroidal neovascularization in high myopia, or idiopathic choroidal neovascularization. Retinal neovascular diseases are all accompanied by the development of retinal non-perfusion areas (NPA), also known as avascular areas (AVA), which are the underlying cause of disease reversal, recurrence, and the need for repeated treatment. Pathological retinal neovascularization is seen in common and severe retinal diseases of all ages. Currently, patients with advanced ROP, PDR, or RVO receive retinal photocoagulation or anti-vascular endothelial growth factor (VEGF) therapy to inhibit neovascularization. Due to the loss of retinal neural tissue caused by the laser, retinal photocoagulation can lead to complications such as decreased vision, decreased night vision, and persistent narrowing of the visual field. Anti-VEGF therapy has been used clinically to treat patients with ROP, PDR, and RVO, but it has potential drawbacks. First, adverse effects associated with blocking vascular endothelial growth factor signaling include impairment of normal retinal vascular growth and retinal function. Second, recurrence of pathological neovascularization is common in premature infants or patients with diabetes after intravitreal injection of anti-VEGF antibodies due to the persistent ischemic / nonperfusion state.

[0015] Furthermore, the molar ratio of the tetrahedral framework nucleic acid to resveratrol is 1:320, that is, tFNAs:RSV is 250 nmol:80 μmol.

[0016] Furthermore, the method for preparing the tetrahedral framework nucleic acid-resveratrol complex comprises the following steps: mixing the resveratrol solution and the tetrahedral framework nucleic acid, and shaking at 3-5° C. for more than 5-7 hours to obtain the tetrahedral framework nucleic acid-resveratrol complex.

[0017] Preferably, the method for preparing the tetrahedral framework nucleic acid-resveratrol complex comprises the following steps: mixing the resveratrol solution and the tetrahedral framework nucleic acid, and shaking at 4° C. for more than 6 hours to obtain the tetrahedral framework nucleic acid-resveratrol complex.

[0018] Furthermore, the drug preparation can be in any dosage form suitable for retinal administration, including but not limited to one or more of injection, eye drops, liposomes or aerosols.

[0019] Furthermore, the tetrahedral framework nucleic acid-resveratrol complex has a particle size of 10 to 15 nm.

[0020] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides the use of a tetrahedral framework nucleic acid (tFNA)-resveratrol complex in the preparation of a drug for treating retinal neovascular diseases. This invention couples resveratrol molecules to the single-stranded DNA backbone of tFNAs via electrostatic adsorption to form a tetrahedral framework nucleic acid-resveratrol complex (tFNAs-RSV). Research in this invention has shown that the tFNAs-RSV complex, when used to treat retinal neovascular diseases, achieves four goals—compared to those achieved simultaneously with tFNAs or RSV alone—of preventing pathological retinal neovascularization, improving retinal perfusion, promoting physiological retinal vascular remodeling, and protecting the retinal neurovascular unit (RNVU). This synergistic effect provides a breakthrough solution for the precise treatment of retinal neovascular diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The synthesis process of tFNAs-RSV and the results of its physicochemical properties analysis are shown below. Figure 1 A is the chemical structure of the small molecule polyphenol resveratrol (RSV); B is a schematic diagram of the synthesis process of tFNAs-RSV; C is the polyacrylamide gel electrophoresis (PAGE) result; D is the high-performance capillary electrophoresis (HPCE) result; E is the Zeta potential analysis result; F is the dynamic light scattering (DLS) result; G is the transmission electron microscopy (TEM) observation result (scale bar: 50 nm).

[0022] Figure 2 The statistical results of the retinal neovascularization area and the avascular area of ​​each group of mice after each drug group was administered to the mouse oxygen-induced retinopathy (OIR) model. Figure 2 A is a quantitative analysis of retinal vessels stained with IB4; B is an enlarged image of localized retinal neovascularization and the area of ​​neovascularization; C shows the retinal avascular zone and the area of ​​avascular zone; D shows the area of ​​venous sprouting and the area of ​​sprouting; E shows the density of vascular network (red) and the density of junctions (blue) (scale bar: 20 μm). The control group, RSV group, tFNAs group, and tFNA-RSV group were included (data are expressed as mean ± SD, n = 6).

[0023] Figure 3 The following are the statistical results of the peak time and amplitude of the electroretinogram b wave of each group of mice after each drug group was administered to the mouse oxygen-induced retinopathy (OIR) model. Figure 3A shows the dark-adapted ERG (1.0 cd·s / m²); B shows the light-adapted ERG (10.0 cd·s / m²); and C shows the results of oscillatory potentials (OPs) analysis. Data are presented as mean ± standard deviation (SD). One-way analysis of variance (ANOVA) was used to compare the two groups (n = 6), and no differences in peak timing were observed. ERG: electroretinogram; OPs: oscillatory potentials. DETAILED DESCRIPTION

[0024] The present invention will be further described below with reference to the accompanying drawings and specific examples, but the examples do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0025] Unless otherwise specified, all reagents and materials used in the following examples were commercially available.

[0026] Example 1 Synthesis of Tetrahedral Framework Nucleic Acid-Resveratrol Complex (tFNAs-RSV) by Electrostatic Adsorption 1. Method (1) Preparation of tFNAs: Shanghai Sangon Biotechnology Co., Ltd. was commissioned to synthesize four single-stranded DNAs (ssDNAs) according to the sequences in Table 1. TM buffer (pH 8.0) containing 10 mM Tris-HCl and 50 mM MgCl₂ was prepared. Equal concentrations of the four ssDNAs were added to the TM buffer, thoroughly mixed, and centrifuged. The mixture was then heated to 95°C for 10 minutes and then cooled to 4°C for 20 minutes to synthesize tetrahedral framework nucleic acids (tFNAs).

[0027] Table 1 Single-stranded DNA sequences for synthesizing tFNAs

[0028] (2) Preparation of tFNAs-RSV: Use a 2 mL EP tube to take an appropriate amount of RSV powder (RSV chemical structure is as follows Figure 1 (as shown in A) was dissolved in DMSO to a concentration of 8000 μmol·L -1 RSV solution, designed the appropriate final concentration of the drug based on previous studies (tFNAs: 250 nmol·L -1 , RSV: 80 μmol·L -1), the RSV solution was mixed with the tFNAs from step (1), placed in a dark environment at 4°C, and shaken for more than 6 hours to allow the tFNAs to fully electrostatically adsorb RSV molecules to obtain a tFNAs-RSV solution, and then ultrafiltered at 4°C and 6000g centrifugal force for 5 minutes to obtain a tFNAs-RSV complex.

[0029] The physicochemical properties of tFNAs-RSV were examined using polyacrylamide gel electrophoresis, high-performance capillary electrophoresis, dynamic light scattering particle size analyzer, potentiometric detection technology, and transmission electron microscopy.

[0030] 2. Results The experimental results are as follows Figure 1 As shown, Figure 1 Figure B is a schematic diagram of the synthesis process of tFNAs-RSV: four specific single-stranded DNAs are designed, assembled under appropriate conditions to form tetrahedral framework nucleic acids (tFNAs), and incubated with RSV to form a complex. Figure 1 Center C shows that the migration bands of tFNAs and tFNAs-RSV were consistent with the theoretical structure, indicating that the complex was successfully constructed. Figure 1 D in the middle is high-performance capillary electrophoresis (HPCE) to further verify its assembly effect, and the results are consistent with PAGE. Figure 1 E in the middle is the Zeta potential analysis, and the results showed that tFNAs and tFNAs-RSV were -8.5 mV and -10.7 mV, respectively, indicating that the complex has good colloidal stability. Figure 1 Figure F shows a slight increase in the particle size of the complex (from 9.1 nm to 12.4 nm) as measured by dynamic light scattering (DLS), indicating that the complex still maintains high structural stability after RSV loading. Figure 1 Medium-G transmission electron microscopy (TEM) observations showed that tFNAs-RSV was in a uniform triangular shape with a complete structure, further verifying that it had a highly stable three-dimensional configuration.

[0031] Example 2 In vivo model experiment The tFNAs-RSV complex prepared in Example 1 was used to observe retinal blood vessels in an oxygen-induced retinopathy (OIR) mouse model. The experimental animals were C57BL / 6 mice. The specific steps included the following: (1) Mice were placed in an oxygen chamber with an oxygen concentration of 75% from the 7th day (P7) to the 12th day (P12) after birth. High oxygen concentrations can lead to the loss of immature retinal blood vessels and slow down the development of the normal retinal vascular system, resulting in the formation of avascular areas in the central retina. (2) At P12, the rats were returned to normal room air (oxygen concentration of approximately 21%). The hypoxic environment induced the expression of angiogenic factors, leading to the regeneration of normal retinal blood vessels and the pathological formation of new blood vessels, simulating the second stage of ROP. (3) P12 intravitreal administration, the administration method is as follows: ① Blank control group (OIR-CTRL): 1 μL normal saline was injected into the vitreous cavity of both eyes; ② Resveratrol monotherapy group (OIR-RSV): 1 μL of 80 μM RSV solution (solvent: normal saline) was administered into the vitreous cavity of both eyes; ③ Tetrahedral framework nucleic acid single-drug group (OIR-tFNAs): 1 μL of 250 nM tFNA solution (solvent: normal saline) was administered into the vitreous cavity of both eyes; ④ tFNAs-RSV complex group (OIR tFNAs-RSV): 1 μL of 250 nM tFNA-RSV complex solution (using the tFNAs-RSV prepared in Example 1, with physiological saline as the solvent) was administered into the vitreous cavity of both eyes; (4) On the 17th day (P17), the retina was observed and immunofluorescence stained, and then photographed. The area of ​​retinal neovascularization and avascular area was calculated using ImageJ.

[0032] result: This example was conducted in a C57BL / 6J mouse OIR model. On postnatal day 12, a control group, RSV (80 μM), tFNAs (250 nM), or tFNAs-RSV (250 nM) were injected intravitreally. The efficacy was evaluated on day 17. IB4-stained retinal flat-mount specimens were analyzed for overall neovascularization (NVA) area. Figure 2 As shown in Figure A, tFNAs-RSV administration significantly reduced neovascularization, significantly better than the tFNAs alone group and the control group. tFNAs-RSV significantly reduced the area of ​​pathological neovascularization to 5.79% ± 2.44% (30.67% ± 3.27% in the control group, P < 0.0001). The results of the enlarged image of the neovascularization area and the quantitative analysis of the area are shown in Figure 2. Figure 2 As shown in Figure B, the above results are further clarified. The non-perfused area (NPA) is demarcated by the yellow dotted circle. Figure 2 As shown in center C, the tFNAs-RSV group had the smallest NPA, indicating a significant restoration of retinal perfusion. The nonperfused area was significantly reduced to 5.09% ± 4.81% (compared to 20.91% ± 2.17% in the control group, P < 0.0001). High-magnification images of vascular sprouting at venous branches and quantitative analysis of the sprouting area are shown in Figure 2. Figure 2As shown in D, tFNAs-RSV significantly promoted the sprouting of new blood vessels at the venous end (44.21% ± 6.38%, compared with 11.49% ± 3.72% in the control group, P < 0.0001). Compared with the RSV group and the tFNAs group, the venous sprouting ability of the tFNAs-RSV group was significantly improved, indicating that tFNAs-RSV has a strong effect in promoting physiological reconstruction. Angio Tool analysis of vascular network structure Figure 2 As shown in Figure E, the tFNAs-RSV group increased overall vascular density (33.26% ± 3.14% vs. 22.78% ± 1.39% in the control group, P < 0.0005) and enhanced vascular network connectivity (connection point density 0.54 ± 0.05 vs. 0.33 ± 0.03 in the control group, P < 0.005). The vascular network connection point density in the tFNAs-RSV group was significantly greater than that in the RSV-only group and the control group. This suggests that tFNAs-RSV has a significant effect on remodeling normal retinal vasculature. This suggests that the tFNAs-RSV complex can simultaneously inhibit pathological neovascularization and reduce the area of ​​retinal non-irrigated areas, promoting retinal neurophysiological vascular remodeling while enhancing vascular network stability, resulting in a synergistic effect.

[0033] Example 3 In vivo model experiment (1) Observe the electroretinogram (ERG) manifestations in the oxygen-induced retinopathy (OIR) mouse model.

[0034] Experimental animals: C57BL / 6 mice; (2) The construction of the OIR mouse model and the intravitreal administration method were the same as in Example 2.

[0035] (3) Start the Celeris mouse electrophysiology platform (Diagnosys, USA) according to the instructions and select the mouse five-step electroretinography program.

[0036] (4) Preparation of P17 mice: The mice were dark-adapted for 12 hours in the evening before the experiment. On the day of the experiment, they were anesthetized by intraperitoneal injection of 0.5% sodium pentobarbital at a dose of 25-30 mg / kg according to their body weight. Tropicamide eye drops were used to dilate the pupils and hydroxymethylcellulose gel was used to moisten the eyes.

[0037] (5) Connect the electrophysiological instrument according to the instrument instructions. Place the mouse flat on the testing platform with its body and limbs fully extended to reduce interference from the myoelectric signal. Fully expose the mouse's eyeballs, and place the left and right stimulators in contact with but not compressing the mouse's cornea.

[0038] (6) Test the connection impedance and adjust the contact angle and position so that the connection impedance is less than 5.0K.

[0039] (7) According to the standard procedures of the International Society for Clinical Electrophysiology of Vision (ISCEV), the following procedures were performed: ① dark adaptation 0.01 ERG, ② dark adaptation 1.0 ERG, ③ dark adaptation 10.0 ERG, ④ dark adaptation oscillatory potential; light adaptation 10 min; ⑤ light adaptation 3.0 ERG, ⑥ light adaptation 30 Hz flicker ERG; before the light adaptation test, the mice received 10 minutes of light adaptation.

[0040] (8) Record the experimental results. Apply tobramycin ointment to protect the eyes of mice and use electric blanket to resuscitate the mice.

[0041] (9) Turn off the computers one by one and clean the laboratory table.

[0042] result: P17 OIR model mice were used to detect dark-adapted ERG, light-adapted ERG and oscillatory potentials (OPs) in different treatment groups, and the grouping was the same as in Example 2. The results of dark-adapted ERG (1.0 cd·s / m²) are as follows: Figure 3 As shown in Figure A, the tFNAs-RSV group had the highest b-wave amplitude, and there was no significant difference in peak time between the groups, indicating that photoreceptor function was best preserved. Figure 3 As shown in Figure B, the b-wave amplitude of the tFNAs-RSV group was significantly higher than that of the CTRL and RSV groups, and there was no significant difference in peak time, which is the same as above. It especially reflects that cones, as the main photoreceptor cells, have the best functional preservation. Figure 3 As shown in middle C, the amplitude of the tFNAs-RSV group was significantly higher than that of the RSV group, but there was no significant difference between the CTRL and tFNAs groups, indicating that the function of bipolar cells was best preserved.

[0043] Retinal electrophysiological function is an important indicator for assessing visual health, directly reflecting the ability of retinal neurons to respond to light stimulation. This study shows that tFNAs-RSV has a significant protective effect on retinal function in an oxygen-induced retinopathy (OIR) model. In full-field ERG testing performed at P17, the tFNAs-RSV group exhibited significantly higher amplitudes in both the scotopic b-wave and the photopic b-wave, increasing by 82% and 85% compared to the control group, respectively. This indicates that the system has a good protective effect on both rod and cone cell function, helping to maintain both scotopic and photopic vision. Furthermore, in terms of oscillatory potentials (OPs), a marker of inner retinal activity, the tFNAs-RSV group showed a significantly better response than the RSV monotherapy group, suggesting its positive significance in improving information transmission and maintaining neural network function. Overall, tFNAs-RSV demonstrates superior and comprehensive therapeutic potential in terms of functional protection. It has a quadruple synergistic mechanism of combating retinal pathological neovascularization, improving retinal perfusion, promoting retinal physiological vascular reconstruction, and protecting the retinal neurovascular unit (RNVU). It achieves the four goals of combating retinal pathological neovascularization, improving retinal perfusion, promoting retinal physiological vascular reconstruction, and protecting the retinal neurovascular unit (RNVU), which are difficult to achieve simultaneously with the single use of tFNAs or RSV. It plays a synergistic role and provides a breakthrough solution for the precise treatment of retinal neovascular diseases.

Claims

1. Use of a tetrahedral framework nucleic acid-resveratrol complex in the preparation of a drug for treating retinal neovascular diseases, characterized in that: The tetrahedral framework nucleic acid-resveratrol complex is a complex formed by uniformly mixing the tetrahedral framework nucleic acid and resveratrol; the molar ratio of the tetrahedral framework nucleic acid to resveratrol is 1:(300-350).

2. The application according to claim 1, characterized in that The tetrahedral framework nucleic acid is formed by four single-stranded DNAs through base complementary pairing, and the nucleotide sequences of the four single-stranded DNAs are shown in SEQ ID No. 1 to 4 respectively.

3. The application according to claim 2, characterized in that: The preparation method of the tetrahedral framework nucleic acid is as follows: four single-stranded DNAs are maintained in a buffer at 85-105° C. for 5-15 minutes, and then maintained at 2-8° C. for 10-30 minutes.

4. The application according to claim 1, characterized in that The drug achieves treatment by resisting retinal pathological neovascularization, improving retinal perfusion, promoting retinal physiological vascular reconstruction, and protecting retinal neurovascular units.

5. The application according to claim 1, characterized in that: The retinal neovascular disease is selected from one or more of diabetic retinopathy, retinopathy of prematurity, retinal vein occlusion, retinal periphlebitis, age-related macular degeneration, polypoidal choroidal vasculopathy, high myopia choroidal neovascularization or idiopathic choroidal neovascularization.

6. The application according to claim 1, characterized in that: The molar ratio of the tetrahedral framework nucleic acid to resveratrol is 1:

320.

7. The use according to claim 1, characterized in that The preparation method of the tetrahedral framework nucleic acid-resveratrol complex comprises the following steps: mixing resveratrol solution and tetrahedral framework nucleic acid, shaking at 2-8° C. for more than 5-7 hours, and obtaining the tetrahedral framework nucleic acid-resveratrol complex.

8. The application according to claim 1, characterized in that: The preparation of the drug is selected from one or more of injection, eye drops, liposomes or aerosols.

9. The use according to claim 1, characterized in that: The particle size of the tetrahedral framework nucleic acid-resveratrol complex is 10 to 15 nm.

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