A dual-tropic peptide-modified nano-particle containing a sting inhibitor specifically targeting vascular endothelium and its preparation method and application

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to a double-membrane peptide modified nano-particle containing a STING inhibitor and specifically targeting vascular endothelium, and a preparation method and application thereof. The STING inhibitor is wrapped in a liposome to obtain a liposome nano-drug, and the liposome as a drug carrier can improve water solubility and bioavailability thereof. Meanwhile, the liposome nano-drug is co-modified by a cell penetrating peptide and a targeting penetrating peptide, and the two can synergistically improve penetrating performance, help the drug to penetrate an ocular barrier, target retinal vascular endothelial cells, and increase bioavailability of the drug. Moreover, the obtained nano-particle exists in a liquid form, and can be directly used as eye drops for treating eyes with retinal diseases. Such a convenient and non-invasive treatment method for eye diseases can better benefit patients with retinal diseases, reduce retinal neovascularization of the patients, and be beneficial to treatment and prognosis of the diseases.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine. More specifically, it relates to a double-transmembrane peptide modified nano-particle containing a STING inhibitor specifically targeting blood vessel endothelium, and a preparation method and application thereof. BACKGROUND

[0002] Retinal neovascularization is the main pathological feature of ischemic retinopathy such as proliferative diabetic retinopathy, retinopathy of prematurity and other diseases, which can lead to vitreous hemorrhage, retinal detachment and even blindness, and has become one of the main causes of blindness in adults and infants. Current treatment methods, such as laser photocoagulation and intravitreal injection of anti-angiogenic factors, are invasive, expensive and ineffective, and all have certain side effects. Although panretinal photocoagulation has achieved short-term success, ocular hemorrhage and visual field defects are ultimately unavoidable. Intravitreal injection of anti-angiogenic factors is widely used to counteract neovascularization and vascular leakage. However, frequent intravitreal injections can also cause complications such as high intraocular pressure. Therefore, there is an urgent need to explore convenient and non-invasive treatment methods such as eye drops to improve treatment efficacy.

[0003] Interferon Stimulatory Factor (STING) was first discovered in the innate immune response. Cyclic GMP-AMP synthase (cGAS) detects abnormal double-stranded DNA accumulation and stimulates the activation of STING, which translocates from the endoplasmic reticulum to the Golgi apparatus, recruits and phosphorylates TANK-binding kinase 1 (TBK1), and then activates IFN regulatory factor 3 (IRF3) to induce the transcription of downstream inflammatory factors. Activation of STING has been shown to be associated with adaptive inflammatory diseases and pathological conditions, including obesity, alcoholic liver disease and cancer.

[0004] More and more evidence reports that STING is closely related to vascular diseases. STING is found to be enriched in vascular endothelial cells in proliferative membrane samples of diabetic retinopathy patients, and the expression level of endothelial cell STING can also be observed in the OIR and normal mouse retina, ranking second only to microglia cells. At the same time, in the OIR mouse retina, the expression of pro-angiogenic factor-related genes is positively correlated with STING. This suggests that targeting and inhibiting endothelial cell STING may help reduce retinal neovascularization. However, current STING inhibitors such as C-176 are mostly injected systemically, lack tissue specificity, and due to the hydrophobicity of C-176, drug delivery is also difficult.

[0005] Despite the multiple protective mechanisms of the eye, such as the corneal barrier, blood-aqueous barrier and blood-retinal barrier, which strictly regulate the exchange and flow of substances in the eye, they also pose significant challenges to drug treatment of eye diseases, especially eye drops for retinal diseases. Given the significant advantages of nanotechnology in drug delivery, research on treatment of eye diseases has gradually attracted attention. However, methods that can achieve non-invasive delivery and achieve good therapeutic effect are still rare.

[0006] Cell-penetrating peptides are considered a potential solution due to their ability to penetrate cell membranes. They can enhance the ability of nanoparticles to cross cell membranes, thereby increasing drug concentration in cells and improving treatment outcomes. Furthermore, targeted cell-penetrating peptides not only have membrane penetration ability, but also specifically recognize and bind to receptors on the surface of target cells, enabling precise targeted drug delivery, thereby more effectively improving treatment outcomes and reducing side effects. For example, existing technologies use targeted cell-penetrating peptides as targeting ligands to modify nanocarriers, enabling active targeting, and can simultaneously carry chemotherapeutic drugs and therapeutic genes to co-treat tumors, thereby increasing drug concentration at the tumor site and improving treatment outcomes.

[0007] However, although targeted cell-penetrating peptides exhibit some targeting ability compared to ordinary cell-penetrating peptides, their targeting effect may still not be ideal. Specifically, some targeted cell-penetrating peptides for tumors may still undergo some degree of non-specific binding in the tumor microenvironment, leading to some drugs being taken up by non-target cells, thereby reducing drug enrichment in target cells and affecting treatment outcomes.

[0008] In summary, for the treatment of retinal diseases, there is an urgent need to develop more efficient, convenient and non-invasive methods to penetrate eye barriers, achieve precise drug delivery, and effectively reduce retinal neovascularization. SUMMARY

[0009] The technical problem to be solved by the present application is to overcome the challenges posed by the multiple protective mechanisms of the eye in drug treatment in the prior art, as well as the defects and shortcomings of targeted cell-penetrating peptides, such as poor targeting effect and difficulty in precise drug release at the target site, and to provide a dual cell-penetrating peptide-modified nanoparticle containing a STING inhibitor.

[0010] The purpose of the present application is to provide a preparation method of the dual cell-penetrating peptide-modified nanoparticle containing a STING inhibitor.

[0011] Another purpose of the present application is to provide applications of the dual cell-penetrating peptide-modified nanoparticle containing a STING inhibitor.

[0012] The above purposes of the present application are achieved by the following technical solutions:

[0013] The application protects a double-penetratin modified STING inhibitor-containing nanoparticle, which is obtained by co-modifying a STING inhibitor-loaded liposome drug with a cell-penetrating peptide and a targeting-penetrating peptide.

[0014] The application wraps a hydrophobic STING inhibitor in a liposome to obtain a liposome nanoparticle drug, the liposome has a phospholipid bilayer structure, and the hydrophobic drug is wrapped in the phospholipid layer. Using the liposome as a drug carrier can improve its water solubility and bioavailability. At the same time, the liposome nanoparticle drug is co-modified with a cell-penetrating peptide and a targeting-penetrating peptide. The cell-penetrating peptide has strong cell penetration, which can help the drug-carrying particles efficiently penetrate the ocular barrier. The targeting-penetrating peptide can specifically recognize and bind to the alphaVbeta3 integrin receptor on the vascular endothelial cells, which is highly expressed on the newly formed vascular endothelial cells, and can achieve drug delivery to the retinal vascular endothelium with good permeability and targeting. The two synergistically improve the penetration performance, help the drug penetrate the ocular barrier, target the retinal vascular endothelial cells, and increase the bioavailability of the drug.

[0015] Further, the STING inhibitor includes one or more of STING C-176, STING-IN-2, STING-IN-3, STING-IN-4, STING-IN-6, STING-IN-7, and STING-IN-8. The above STING inhibitors can all achieve the effect of treating retinal vascular diseases by inhibiting the STING pathway.

[0016] Further, the amino acid sequence of the cell-penetrating peptide is shown in SEQ ID NO. 1 (GRKKPRQRRRPPQ).

[0017] Further, the cell-penetrating peptide includes a cell-penetrating peptide-polymer conjugate. When the cell-penetrating peptide is a cell-penetrating peptide-polymer conjugate, the conjugate can enhance the stability and dispersibility of the particles, reduce particle aggregation, enhance the biocompatibility of the particles, and reduce the probability of being cleared by the immune system.

[0018] Further, the cell-penetrating peptide-polymer conjugate includes a phospholipid-polyethylene glycol-cell-penetrating peptide.

[0019] Further, the phospholipid-polyethylene glycol-cell-penetrating peptide is synthesized by Michael addition of a phospholipid-polyethylene glycol-maleimide (DSPE-PEG-Mal) and a cell-penetrating peptide.

[0020] Further, the targeting-penetrating peptide is an integrin-targeting cyclic peptide.

[0021] Further, the amino acid sequence of the targeting cell-penetrating peptide is shown as SEQ ID NO. 2 (CRGDKGPDC).

[0022] Further, the targeting cell-penetrating peptide comprises a targeting cell-penetrating peptide-polymer conjugate, and the targeting cell-penetrating peptide-polymer conjugate comprises a phospholipid-polyethylene glycol-integrin targeting cyclic peptide. When the targeting cell-penetrating peptide is a targeting cell-penetrating peptide-polymer conjugate, the connection of the conjugate can enhance the stability and dispersibility of the particles, reduce particle aggregation, enhance the biocompatibility of the particles, and reduce the probability of being cleared by the immune system.

[0023] Further, the targeting cell-penetrating peptide-polymer conjugate comprises a phospholipid-polyethylene glycol-integrin targeting cyclic peptide iRGD (DSPE-PEG-iRGD).

[0024] Further, the raw material of the liposome in the STING inhibitor-loaded liposome nanoparticle comprises one or more of a phospholipid, a phospholipid-polymer conjugate, and cholesterol.

[0025] Further, the phospholipid is one or more of a negatively charged phospholipid, a positively charged phospholipid, and a neutral phospholipid.

[0026] Further, the negatively charged phospholipid comprises one or more of phosphatidylglycerol, phosphatidic acid, and phosphatidylinositol.

[0027] Further, the positively charged phospholipid comprises a phosphatidylcholine cholesteryl derivative.

[0028] Further, the phospholipid-polymer conjugate comprises distearoylphosphatidyl ethanolamine-polyethylene glycol.

[0029] The present application also protects a preparation method of the double cell-penetrating peptide modified STING inhibitor-containing nanoparticle, comprising the following steps:

[0030] S1. Mix the liposome raw material, the targeting cell-penetrating peptide, the cell-penetrating peptide, and the STING inhibitor with an organic solvent to obtain a mixture; remove the solvent from the obtained mixture to prepare a lipid membrane, and then perform hydration treatment to obtain a liposome dispersion; and sufficiently disperse the obtained liposome dispersion to obtain the double cell-penetrating peptide modified STING inhibitor-containing nanoparticle.

[0031] The double membrane peptide modified STING inhibitor containing nanoparticles prepared by the above preparation method exist in liquid form, which can be directly used as eye drops for treating retinal diseases. After administration, the activation of the retinal STING pathway is weakened, the pathological neovascularization of the retina is reduced, the vascular tortuosity is reduced, the retinal inflammation is reduced, and the perivascular cell coverage is improved. This convenient and non-invasive eye disease treatment method can better benefit patients with retinal diseases, reduce neovascularization, and is beneficial to the treatment and prognosis of diseases.

[0032] Further, the molar ratio of the liposome raw material, the targeting cell-penetrating peptide and the cell-penetrating peptide is 1:(0.01-0.1):(0.01-0.1), preferably 1:(0.03-0.08):(0.03-0.08).

[0033] Further, the ratio of the mass of the STING inhibitor to the total mass of the liposome raw material, the targeting cell-penetrating peptide and the cell-penetrating peptide is 1:(10-30), preferably 1:(15-25).

[0034] Further, the organic solvent includes one or more of chloroform, diethyl ether, and methanol.

[0035] Further, the solid-liquid ratio of the raw material is 1:(200-300) g / mL, preferably 1:(220-250) g / mL. In the solid-liquid ratio, the "solid" refers to a solid, and the solid includes materials such as liposome raw materials, targeting cell-penetrating peptides, cell-penetrating peptides, and STING inhibitors. The "liquid" in the solid-liquid ratio refers to a liquid, and the liquid is an organic solvent.

[0036] Further, the raw material of the liposome in the STING inhibitor loaded liposome nanoparticle includes one or more of phospholipids, phospholipid-polymer conjugates, and cholesterol.

[0037] Further, the phospholipid is one or more of negatively charged phospholipids, positively charged phospholipids, and neutral phospholipids.

[0038] Further, the negatively charged phospholipid includes one or more of phosphatidylglycerol, phosphatidic acid, and phosphatidylinositol.

[0039] Further, the positively charged phospholipid includes phosphatidylcholine cholesterol derivatives.

[0040] Further, as an optional embodiment, the raw material of the liposome is phospholipid and cholesterol, and the molar ratio of the cholesterol and the phospholipid is 1:(5-12), more preferably 1:(7-9).

[0041] Further, the method for removing the solvent to form a lipid film is rotary evaporation.

[0042] Further, the conditions of the rotary evaporation film formation are 35-40℃, 100-150rpm, and 10-20min.

[0043] Further, the hydration treatment is to add water to the system to strip the lipid film, and the lipid film is hydrated and peeled off to produce a liposome dispersion.

[0044] Further, the sufficient dispersion method is ultrasonic treatment, and the conditions of the ultrasonic treatment are 100-150w, 20-30kHz, and 15-25min.

[0045] The application also protects the use of the double-membrane peptide modified STING inhibitor-containing nanoparticle in the preparation of an anti-retinopathy drug.

[0046] Further, the retinopathy includes retinal vascular disease.

[0047] Further, the anti-retinopathy includes reducing retinal inflammation and reducing retinal neovascularization.

[0048] Compared with the prior art, the application has the following beneficial effects:

[0049] The application wraps the STING inhibitor in liposomes to obtain a liposome nanoparticle drug, and the liposome as a drug carrier can improve the water solubility and bioavailability thereof; at the same time, the liposome nanoparticle drug is co-modified by a cell-penetrating peptide and a targeting cell-penetrating peptide, and the two synergistically improve the cell-penetrating performance, help the drug penetrate the ocular barrier, target the retinal vascular endothelial cells, and increase the bioavailability of the drug. Moreover, the obtained nanoparticle exists in a liquid state, and can be directly used as eye drops for treating the eyes with retinopathy. Such a convenient and non-invasive eye disease treatment method can better benefit the patients with retinal diseases, reduce the retinal neovascularization of the patients, and be conducive to the treatment and prognosis of the diseases. BRIEF DESCRIPTION OF DRAWINGS

[0050] Figure 1 The figure is a schematic diagram of the iRGD&TAT-C176-NP nanoparticle.

[0051] Figure 2 The figure is a morphological diagram of iRGD-C176-NP and iRGD&TAT-C176-NP under a transmission electron microscope (TEM).

[0052] Figure 3 The figure is a data statistical diagram of the drug release rate of iRGD-C176-NP and iRGD&TAT-C176-NP.

[0053] Figure 4Statistical graph of the efficiency of iRGD-Nile red-NP and iRGD&TAT-Nile red-NP into the retina after 6 hours of eye drops.

[0054] Figure 5 Statistical graph of the results of the scratch test of iRGD-C176-NP and iRGD&TAT-C176-NP on endothelial cells.

[0055] Figure 6 Schematic diagram of eye drops treatment of OIR mice.

[0056] Figure 7 Statistical graph of p-TBK1 staining of retinal flat mounts of OIR mice after 4 days of continuous eye drops.

[0057] Figure 8 Fundus angiography of mice after 4 days of continuous eye drops.

[0058] Figure 9 Statistical graph of retinal vascular flat mounts and the area of new blood vessels and non-concerned areas of OIR mice after 4 days of continuous eye drops.

[0059] Figure 10 Statistical graph of retinal glial fibrillary acidic protein (GFAP) staining of mice after 4 days of continuous eye drops.

[0060] Figure 11 Statistical graph of retinal neuroglia antigen 2 (NG2) staining of mice after 4 days of continuous eye drops. DETAILED DESCRIPTION

[0061] The present application will be further described by the following description of drawings and specific examples, but the examples do not limit the present application in any form. Unless otherwise specified, the reagents, methods and equipment used in the present application are conventional reagents, methods and equipment in the technical field.

[0062] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0063] Statistical analysis of example data: all results are expressed as mean ± standard deviation. Comparison between two groups was performed using paired t test, and comparison between multiple groups was performed using one-way ANOVA. *P<0.05, **P<0.01, ***P<0.001 were considered statistically significant.

[0064] Example 1 Preparation of a double TAT-modified STING inhibitor-containing nanoparticle specifically targeting vascular endothelium

[0065] 1. Experimental method

[0066] ①Material preparation: Cholesterol and phospholipid were purchased from Avanti (Shanghai) Co., Ltd. Phospholipid-polyethylene glycol-integrin targeting cyclic peptide iRGD (DSPE-PEG-iRGD, its amino acid sequence is CRGDKGPDC) was purchased from Guangzhou Carbohydrate Technology Co., Ltd. Phospholipid-polyethylene glycol-maleimide (DSPE-PEG-Mal) was purchased from Shanghai Bide Pharmaceutical Technology Co., Ltd. Phospholipid-polyethylene glycol-cell penetrating peptide TAT (DSPE-PEG-TAT) was synthesized by Michael addition of DSPE-PEG-Mal and TAT sequence (GRKKPRQRRRPPQ). STING inhibitor C-176 was purchased from MedChemExpress Biotechnology Co., Ltd. in the United States. Nile red was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0067] ②Preparation by thin film hydration method: 40 mg of cholesterol: phospholipid: DSPE-PEG-iRGD: DSPE-PEG-TAT (molar ratio 1:8:0.5:0.5) and 2 mg of STING inhibitor (C-176) were dissolved in 10 ml of chloroform. The chloroform was removed by rotary evaporation (35 degrees Celsius, 135 revolutions per minute, 20 minutes), and then a lipid film was prepared. Then, 4 ml of ultrapure water was added to peel off the lipid film, and the desired double penetrating peptide modified STING inhibitor containing specific targeting vascular endothelial nanoparticles (named iRGD&TAT-C176-NP) were obtained by ultrasonic treatment (130 watts, 20 kilohertz, 20 minutes). The solution without TAT was prepared with cholesterol: DSPE-PEG-iRGD: DSPE-PEG-Mal (molar ratio 1:8:0.5:0.5), and the remaining steps and parameters were the same as above to obtain iRGD-C176-NP nanoparticles without TAT. The iRGD&TAT-C176-NP or iRGD-C176-NP obtained above can be referred to as eye drops. The preparation method of Nile red-loaded liposome particles iRGD&TAT-C176-Nile red-NP and iRGD-C176-Nilered-NP is the same as above.

[0068] 2、Experimental results

[0069] The structure diagram of iRGD&TAT-C176-NP nanoparticles is shown in Figure 1 , and the STING inhibitor C-176 is encapsulated in the liposome, which is modified by cell penetrating peptide TAT and targeting penetrating peptide iRGD.

[0070] The transmission electron microscopy diagram of iRGD&TAT-C176-NP nanoparticles is shown in Figure 2 , and the nanoparticles present a spherical structure.

[0071] Example 2 Drug Release Detection

[0072] 1. Experimental Methods

[0073] A dialysis bag (MWCO: 3500 Da) containing 1 mL of eye drops (iRGD&TAT-C176-NP or iRGD-C176-NP) was dialyzed in 8 mL of PBST (PBS containing 0.1% Tween-20) at 37°C. Then, after 0, 0.5, 1, 2, 4, 8, and 12 hours, 1 mL of culture medium was removed and the same volume of PBST was added. The concentration of C-176 was determined by high-performance liquid chromatography (HPLC).

[0074] 2. Experimental Results

[0075] Drug release results of iRGD&TAT-C176-NP nanoparticles and iRGD-C176-NP nanoparticles without TAT are as follows: Figure 3 As shown, approximately 60% was released within 12 hours, with no significant difference in release rate between the two.

[0076] Example 3 Application of nanoparticles modified with double-penetrating membrane peptides and containing STING inhibitors that specifically target vascular endothelium 1. Experimental methods

[0077] (1) Scratch test

[0078] Cell culture: Human retinal vascular endothelial cells (HRVEC) were maintained in DMEM complete medium (1 g / L glucose) containing 10% fetal bovine serum and 1% penicillin and streptomycin, and cultured in a cell culture incubator at 37 degrees Celsius and 5% carbon dioxide.

[0079] Scratch assay: Cells were seeded in 6-well plates and, after reaching 100% confluence, scratches were created using a 200 μL pipette tip. Images were taken at 0 and 16 hours after treatment with eye drops (iRGD&TAT-C176-NP or iRGD-C176-NP) and the scratch width was measured.

[0080] (2) Determination of the oxygen-induced retinopathy (OIR) model

[0081] OIR model: Mouse pups were housed in room air (RA) with 21% oxygen until day 7 (P7), then exposed to 75% oxygen for 5 consecutive days from P7 to P12. Hyperoxia induced retinal vascular occlusion in mice. The animals were then returned to RA for 5 days from P12 to P17. During this relatively hypoxic period, the mouse retina underwent neovascularization, peaking at P17. Age-matched pups were housed in RA as a control group. Before eye drop treatment, mice were randomly assigned to a control group, an iRGD-C176-NP group, or an iRGD&TAT-C176-NP group. Starting at P14, 10 μL of the corresponding eye drops were instilled into each eye daily. Samples were taken 6 hours after eye drops were instilled on P17 for relevant tests.

[0082] (3) Fluorescein fundus angiography

[0083] Fluorescein fundus angiography: 200 mL of 1% sodium fluorescein was injected intraperitoneally into anesthetized mice. Fundus images were acquired using the Micron IV fundus endoscope system (Phoenix Research Labs, USA), and vascular tortuosity was quantified using ImageJ.

[0084] (4) Retinal patching and staining

[0085] Retinal preparation and staining: The retina was fixed in 4% paraformaldehyde for 10 minutes, then transferred to 2X PBS for dehydration for 10 minutes. The retina was then dissected under a microscope and prepared into slides, which were then fixed in pre-cooled 100% methanol. Before staining, retinal slides or frozen retinal sections were removed, washed three times with PBS, and then ruptured with 0.3% Triton solution for 30 minutes. The slides were then incubated overnight at 4°C with isoglucosinolate GS-IB4 (I21411; Invitrogen) and primary antibodies, including anti-p-TBK1 (5483S, CST), anti-GFAP (3670S, CST), and anti-NG2 (AB5320, Merck Millipore). Anti-rabbit and anti-mouse secondary antibodies were incubated at room temperature for 1 hour. After thorough washing, the samples were mounted with a mounting medium (ab104139, Abcam). Images were taken under a laser confocal microscope.

[0086] 2. Experimental Results

[0087] Immunofluorescence staining images of frozen sections of mouse eyeballs 6 hours after eye instillation with iRGD&TAT-C176-Nile red-NP nanoparticles and iRGD-C176-Nile red-N nanoparticles containing Nile Red are shown below. Figure 4 As shown, the fluorescence intensity of the iRGD and TAT dual-modified particle group reaching the retinal blood vessels was significantly higher than that of the iRGD-modified group alone.

[0088] The results of scratch assays on endothelial cells using C-176-encapsulated particles are as follows: Figure 5 As shown, the iRGD & TAT-C176-NP dual-modified particles exhibit a stronger inhibitory effect on endothelial cell migration.

[0089] A schematic diagram of the experiment involving the treatment of OIR mice with eye drops is shown below. Figure 6 As shown, mice were given the corresponding eye drops for four consecutive days starting at P14, and samples were taken 6 hours after the eye drops were given at P17 for relevant tests.

[0090] p-TBK1 staining of retinal vessels in mice at P17 is shown in the image. Figure 7 As shown, p-TBK1 is a downstream activation signal of STING. Its fluorescence intensity in the retinal vessels of mice in the iRGD&TAT-C176-NP group was weak, which means that it exerted a stronger inhibitory effect on STING signaling in vascular endothelium.

[0091] The results of fluorescein fundus angiography at P17 in mice are as follows: Figure 8 As shown, the retinal vascular tortuosity of OIR mice was significantly increased, while the treatment with eye drops encapsulated with C-176 could reduce the vascular tortuosity. The treatment effect of iRGD and TAT dual-modified particles was better.

[0092] Vascular staining results of retinal patch in mice at P17 are as follows: Figure 9 As shown, eye drops encapsulated with C-176 can reduce the area of ​​neovascularization and the area of ​​non-perfusion vascularization, and the therapeutic effect of iRGD and TAT dual-modified particles is better.

[0093] GFAP staining of retinal vessels in mice at P17 is shown in the image. Figure 10 As shown, GFAP-labeled astrocytes can reflect the inflammatory state of the retina. Eye drops encapsulated with C-176 can reduce retinal inflammation, and iRGD and TAT dual-modified particles have better anti-inflammatory effects.

[0094] NG2 staining image of retinal vessels in mice at P17 is shown below. Figure 11 As shown, NG2-labeled pericytes of blood vessels, and the pericyte coverage reflects the maturity and stability of blood vessels. Eye drops encapsulated with C-176 can increase retinal pericyte coverage, and the therapeutic effect is even better with iRGD and TAT dual-modified particles.

[0095] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A double-penetrating membrane peptide-modified nanoparticle containing a STING inhibitor, characterized in that, The STING inhibitor-containing nanoparticles modified with the double-penetrating peptide were obtained by co-modifying a liposomal drug loaded with a STING inhibitor using cell-penetrating peptides and targeting-penetrating peptides. The STING inhibitor is STING C-176; The cell-penetrating peptide is a phospholipid-polyethylene glycol-cell-penetrating peptide conjugate; the amino acid sequence of the cell-penetrating peptide is shown in SEQ ID NO.1; The targeted membrane-penetrating peptide is a phospholipid-polyethylene glycol-integrin targeted cyclic peptide conjugate.

2. The STING inhibitor-containing nanoparticles modified with a double-penetrating membrane peptide according to claim 1, characterized in that, The amino acid sequence of the integrin-targeting cyclic peptide is shown in SEQ ID NO.

2.

3. The method for preparing STING inhibitor-containing nanoparticles modified with double-penetrating membrane peptides as described in claim 1 or 2, characterized in that, Includes the following steps: S1. Liposome raw materials, targeted membrane-penetrating peptides, cell membrane-penetrating peptides, STING inhibitors and organic solvents are mixed to obtain a mixture; the solvent is removed from the obtained mixture to form a lipid membrane, which is then hydrated to obtain a liposome dispersion; the obtained liposome dispersion is fully dispersed to obtain nanoparticles modified with dual-membrane-penetrating peptides and containing STING inhibitors.

4. The preparation method according to claim 3, characterized in that, The molar ratio of the liposome raw material, the targeted membrane-penetrating peptide, and the cell membrane-penetrating peptide is 1:(0.01~0.1):(0.01~0.1).

5. The preparation method according to claim 3, characterized in that, The ratio of the mass of the STING inhibitor to the total mass of the liposome raw material, cell-targeting peptide, and cell-penetrating peptide is 1:(10~30).

6. The preparation method according to claim 3, characterized in that, The organic solvent includes one or more of chloroform, diethyl ether, and methanol.

7. The use of the double-penetrating membrane peptide-modified nanoparticles containing STING inhibitors as described in claim 1 or 2 in the preparation of anti-retinal disease drugs.

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