Pharmaceutical composition of SN-011 and application thereof

Abstract

The invention discloses a medicinal composition of SN-011 and application thereof, the medicinal composition comprises SN-011, cyclodextrin and a solvent, the concentration of SN-011 is 0.5-2 mg / mL, and the molar ratio of SN-011 to cyclodextrin is 1: (30-60); and the cyclodextrin is selected from alpha-cyclodextrin, beta-cyclodextrin, gamma-cyclodextrin, sodium sulfobutyl beta-cyclodextrin and dimethyl beta-cyclodextrin. The pharmaceutical composition for solubilizing the SN-011 is prepared by taking an innovative small molecular compound SN-011 of a targeted STING protein as a starting point and adopting cyclodextrin inclusion, so that the solubility bottleneck of the SN-011 when being applied to a pharmaceutical preparation is broken through, multi-dosage-form achievement industrialization is expected to be realized, and important fundamental research significance is provided for SN-011 in various treatment fields.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a pharmaceutical composition of SN-011 and its application. Background Technology

[0002] SN-011 is a small molecule inhibitor discovered through compound screening and bioactivity evaluation targeting the STING protein. It exhibits potent inhibitory activity against the STING signaling pathway. Mechanistic studies further confirm that SN-011 specifically binds to the pocket where the STING protein binds to its endogenous ligand cGAMP, significantly inhibiting downstream overactivated inflammatory cytokines. Its structural formula is shown below:

[0003]

[0004] SN-011 significantly reduced the expression and secretion of pro-inflammatory factors such as IFN-1 in multiple in vitro and in vivo models, and also inhibited the IC50 of STING signaling. 50 With a concentration of only 76 nmol / L, high efficacy, and low cytotoxicity, previous studies have shown that SN-011 has high drug-likeness. Therefore, SN-011 was selected as a lead compound for optimization and further development into related formulations, showing broad application prospects. SN-011 is almost insoluble in water, with a solubility of less than 0.2 μg / mL in water at 25°C. Therefore, improving the solubility of SN-011 is a challenge in developing clinical dosage forms for this drug.

[0005] For the solubilization of poorly soluble drugs, from the perspective of formulation product development and transformation, adopting simpler formulations, processes, and dosage forms is beneficial for production quality control and cost control, as well as clinical application. Furthermore, formulation development should comprehensively consider the drug's mechanism of action, effective concentration, and clinical application scenarios, while also taking into account the use of excipients already marketed in similar formulations both domestically and internationally, as well as the process and quality control challenges of related dosage forms.

[0006] In summary, there is an urgent need to develop SN-011 solution formulations that have good solubilizing effects and are easy to convert. Summary of the Invention

[0007] One of the objectives of this invention is to provide a pharmaceutical composition of SN-011, comprising SN-011, cyclodextrin and a solvent, wherein the concentration of SN-011 is 0.5 to 2 mg / mL and the molar ratio of SN-011 to cyclodextrin is 1:(30-60).

[0008] The cyclodextrin is selected from α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, sodium sulfobutylether-β-Cyclodextrin (SBE-β-CD), hydroxypropyl-β-cyclodextrin (HP-β-CD), and dimethyl-β-cyclodextrin (DM-β-CD).

[0009] The solvent is water or a mixture of water and an organic solvent.

[0010] Furthermore, the organic solvent is selected from ethanol, propylene glycol, butanediol, isopropanol, glycerol, dimethylacetamide, polyethylene glycol 300, polyethylene glycol 400, and polyethylene glycol 600.

[0011] Furthermore, the volume concentration of the organic solvent in the mixed solvent is 5-50%; preferably 10-30%.

[0012] Furthermore, the cyclodextrin is SBE-β-CD or HP-β-CD.

[0013] Furthermore, the pharmaceutical composition of said SN-011 is selected from one of the following:

[0014] (a) SN-011, SBE-β-CD and water, with SN-011 concentration of 0.5-2 mg / mL and the molar ratio of SN-011 to SBE-β-CD of 1:(30-60);

[0015] (ii) A mixed solvent of SN-011, HP-β-CD, water and organic solvent, wherein the concentration of SN-011 is 0.5 to 2 mg / mL, the molar ratio of SN-011 to HP-β-CD is 1:(30-60), and the volume concentration of the organic solvent in the mixed solvent is 5 to 50%, preferably 10 to 30%.

[0016] Furthermore, in combination (a), a stabilizer may be added, with a concentration of 0.5–2.5 mg / mL. The stabilizer is a water-soluble polymer or a pH adjuster. The water-soluble polymer is selected from polyvinyl alcohol, sodium alginate, chitosan, povidone, and hyaluronic acid. The pH adjuster is an acidic pH adjuster and / or an alkaline pH adjuster.

[0017] Specifically, the acidic pH adjuster is selected from one, two, or more of ascorbic acid, lactic acid, malic acid, fumaric acid, citric acid, tartaric acid, succinic acid, hydrochloric acid, phosphoric acid, and acetic acid; the alkaline pH adjuster may be selected from one, two, or more of sodium hydroxide, sodium carbonate, sodium bicarbonate, triethylamine, diethanolamine, sodium phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, meglumine, arginine, lysine, histidine, and glycine.

[0018] In this invention, the pharmaceutical composition of SN-011 is prepared by one of the following methods:

[0019] Method 1: Dissolve SN-011 in an organic solvent to obtain an SN-011 solution, add an aqueous solution of SBE-β-CD to the SN-011 solution for inclusion, evaporate to remove the organic solvent, and obtain the pharmaceutical composition.

[0020] Method 2: Dissolve SN-011 in an organic solvent to obtain an SN-011 solution, dissolve HP-β-CD in a mixed solvent of water and organic solvent to obtain a cyclodextrin solution, add the cyclodextrin solution to the SN-011 solution for inclusion, evaporate to remove the organic solvent, and obtain the pharmaceutical composition.

[0021] A second objective of this invention is to provide the use of the above-described pharmaceutical composition in the preparation of medicaments for the prevention or treatment of diseases mediated by the STING signaling pathway. The medicament is a pharmaceutically acceptable oral or injectable formulation made from the above-described pharmaceutical composition as the main active ingredient and pharmaceutically acceptable pharmaceutical excipients.

[0022] In one embodiment of the present invention, an injection of SN-011 is provided, with the above-described pharmaceutical composition as the active ingredient, and further comprising pharmaceutically acceptable excipients. The injection may be a solution or a lyophilized powder for injection. The lyophilization protectant may be one or more of mannitol, trehalose, sucrose, lactose, maltose, sorbitol, and xylitol.

[0023] Diseases mediated by the STING signaling pathway include infectious diseases, inflammatory diseases, autoimmune diseases, organ fibrosis, ischemic cardiovascular and cerebrovascular diseases, degenerative neurological diseases, traumatic brain injury, spinal cord injury, cancer, or cancer syndromes. Alternatively, diseases mediated by the STING signaling pathway include psoriasis, stroke, traumatic brain injury, spinal cord injury caused by trauma or other diseases, Parkinson's disease, Huntington's disease, familial amyotrophic lateral sclerosis, myocardial infarction, non-alcoholic steatohepatitis, systemic lupus erythematosus, Aicardi-Goutières syndrome, rheumatoid arthritis, inflammatory bowel disease, STING-associated vascular disease (SAVI) occurring in infancy, or diabetes and its complications.

[0024] Beneficial effects:

[0025] 1. This invention takes the innovative small molecule compound SN-011 targeting the STING protein as its starting point, and uses cyclodextrin inclusion complexation to prepare a pharmaceutical composition for solubilizing SN-011, breaking through the solubility bottleneck of SN-011, and is expected to realize the industrialization of multi-dosage form results, providing important basic research significance for SN-011 in a variety of therapeutic fields.

[0026] 2. The pharmaceutical composition for solubilizing SN-011 provided by the present invention has a simple and easy-to-operate preparation method, high efficiency, and is suitable for industrial production; it has low organic solvent content, high safety, and high quality stability of the solubilization system, and has great development potential and clinical application value. Attached Figure Description

[0027] Figure 1 This is a schematic diagram of an IMQ-induced psoriasis treatment procedure.

[0028] Figure 2 These are daily morphological images of the skin on the backs of mice in each group during the modeling period.

[0029] Figure 3 PASI scores (Mean±SD, n=5) for each group of mice. Detailed Implementation

[0030] The preferred embodiments of the present invention will now be described in detail with reference to specific examples. It should be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the invention. Those skilled in the art can make various modifications and substitutions to the present invention without departing from its spirit and essence.

[0031] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0032] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0033] The detection conditions for the SN-011 content determination method used in the following examples are shown in the table below:

[0034]

[0035] Exploration of solubilization methods for SN-011

[0036] 1. Co-solvent

[0037] Cosolvents are water-soluble organic solvents widely used to increase the solubility of poorly soluble drugs. Cosolvents have low polarity and reduce solution polarity by disrupting the intermolecular hydrogen bond network present in aqueous systems. Based on this mechanism and the principle of "like dissolves like," cosolvent systems can improve the solubility of nonpolar drugs in water. Commonly used organic solvents include propylene glycol, glycerol, ethanol, dimethylacetamide, polyethylene glycol 400, and polyethylene glycol 300. Drugs lacking ionizing functional groups and with moderate logP (1–3) may be more suitable for solubilization using cosolvents because they have relatively low solubilizing affinity for surfactants or lipids. However, dilution significantly reduces the solubilizing ability of cosolvents, thus introducing the risk of drug precipitation. Therefore, whether precipitation occurs after dilution before administration is a key parameter for evaluating cosolvent formulations. In addition, the compatibility and toxicity of the cosolvent should also be considered.

[0038] The solubility of SN-011 in different solvents is shown in the table below:

[0039]

[0040] The above results indicate that SN-011 has extremely poor solubility in water, and solubilization techniques are needed to develop it into a solution formulation. However, based on the solubility results of SN-011 in common solvents, it is known that SN-011 has higher solubility in PEG-400 and dimethylacetamide (DMA). Therefore, it is possible to increase the solubility of SN-011 by forming a co-solvent system by mixing these two solvents with water.

[0041] Based on the solubility of SN-011 in common solvents, polyethylene glycol 400 and dimethylacetamide were preferred. A search of the FDA-approved drug inactive ingredient database showed that the maximum dosage of polyethylene glycol 400 and dimethylacetamide for intravenous injection was 30% per unit. Therefore, the solubilizing ability of different concentrations of polyethylene glycol 400 and dimethylacetamide solutions, as well as their mixtures, for SN-011 was first investigated. Specifically, the prescribed amounts of polyethylene glycol 400 and dimethylacetamide solutions were added to purified water and mixed thoroughly at room temperature. Then, the prescribed amount of SN-011 was added, and the mixture was magnetically stirred at 600 rpm for 5 hours at room temperature. The mixture was then brought to a final volume with purified water, and the insoluble matter was removed by filtration through a 0.22 μm filter membrane. The appearance of the samples was observed, and the SN-011 content was calculated. The results are shown in the table below.

[0042]

[0043] The above experiments show that when only one organic solvent is used in the system, the solubility of SN-011 increases slowly with the increase of the organic solvent concentration. When the concentrations of polyethylene glycol 400 and dimethylacetamide reach the maximum limits specified in the FDA-approved drug inactive ingredient database, they can only solubilize 8% of the prescribed amount of SN-011, indicating weak solubilizing ability. When two mixed organic solvents are used in the system, they can achieve a latent solubility effect on SN-011, improving the solubilizing effect compared to a single organic solvent. When the concentration of the mixed organic solvent reaches 50%, it can solubilize 80% of the prescribed amount of SN-011. However, excessively high organic solvent concentrations can cause safety issues such as hemolysis and vascular irritation. Furthermore, after dilution, changes in organic solvent concentration can cause exponential changes in drug solubility, leading to the risk of drug precipitation. Therefore, a single latent solvent system is not suitable for solubilizing SN-011.

[0044] 2. Cyclodextrin inclusion complex

[0045] The combined effect of a certain concentration of organic solvent and cyclodextrin can improve drug solubility. However, the interaction mechanism of the organic solvent-drug-cyclodextrin ternary complex system is complex, and different types of cyclodextrin have different solubilizing effects on drugs. Currently, the cyclodextrins used in commercially available cyclodextrin inclusion complex injection products are mostly SBE-β-CD and HP-β-CD. However, SBE-β-CD has poor solubility in many organic solvents, therefore HP-β-CD is preferred when combined with injectable organic solvents.

[0046] According to the inventors' preliminary experimental tests, the solubility of SN-011 in 16% (w / v) aqueous solutions of HP-β-CD and SBE-β-CD at 37℃ was 14.9 μg / mL and 5.4 μg / mL, respectively. The results confirmed that while a single cyclodextrin aqueous solution increased the solubility of SN-011 by nearly 20 times, it was still difficult to reach the prescribed dosage of 1 mg / mL. Therefore, it is possible to consider using an organic solvent to dissolve SN-011, allowing it to enter the cyclodextrin cavity in a molecular state, and finally removing the organic solvent by rotary evaporation. Because HP-β-CD and SBE-β-CD have extremely high solubility in water, conventional inclusion complex preparation methods such as saturated solution methods and grinding methods are not suitable for preparing inclusion complex solutions. The preparation methods for cyclodextrin inclusion complex injections typically employ methods such as solution stirring-evaporation, ultrasonic-evaporation, and solution stirring.

[0047] (1)SBE-β-CD / EtOH

[0048] Weigh an appropriate amount of SBE-β-CD and place it in a round-bottom flask. Add ultrapure water and then place the flask in an oil bath and stir magnetically to dissolve it. Separately weigh SN-011, add an appropriate amount of anhydrous ethanol and dissolve it by sonication. Slowly add the SN-β-CD solution dropwise to the SBE-β-CD solution. Heat and stir, then turn off the heating and continue stirring until the inclusion complex solution cools to room temperature. Then, remove the solvent by vacuum rotary evaporation. Add ultrapure water to hydrate the solution to obtain the final product.

[0049] (2) HP-β-CD / PEG-400

[0050] Weigh out HP-β-CD, add PEG-400 and purified water, and dissolve in an oil bath by stirring to obtain an HP-β-CD solution. Separately weigh out SN-011, add PEG-400, and dissolve in an oil bath by stirring to obtain an SN-011 solution. Add the HP-β-CD solution to the SN-011 solution, heat and stir, then turn off the heating and continue stirring until the inclusion complex solution cools to room temperature to obtain the final product.

[0051] Example 1

[0052] The SN-011 solubilization system was prepared using the following formulation.

[0053] Prescription: SN-011 10mg

[0054] SBE-β-CD 2.75g

[0055] Add water to 10mL

[0056] Preparation method one is as follows:

[0057] Weigh 2.75 g of SBE-β-CD into a 100 mL round-bottom flask, add 4 mL of purified water, and dissolve by magnetic stirring in a 60 °C oil bath. Separately, accurately weigh 10 mg of SN-011 and add it to the SBE-β-CD solution. Heat and stir for 4 h, then turn off the heat and continue stirring for 1 h to allow the inclusion complex solution to cool to room temperature. Transfer the resulting solution to a 10 mL volumetric flask, dilute to the mark with purified water, shake well, and filter through a 0.22 μm filter membrane to remove insoluble matter.

[0058] Finished product testing: Accurately transfer 200 μL of the formulation into a 10 mL volumetric flask, dilute to the mark with methanol, shake well, and let stand to prepare a solution containing approximately 20 μg of the finished product per mL; accurately measure 20 μL and record the chromatogram using high-performance liquid chromatography (HPLC). Separately, take SN-011 active pharmaceutical ingredient and determine it using the same method as a control group. Calculate the product based on peak area using the external standard method.

[0059] The encapsulation efficiency of the inclusion compound is calculated using the following formula:

[0060] Encapsulation rate (%) = (Amount of drug encapsulated / Drug dosage) * 100%

[0061] The calculated encapsulation efficiency of SN-011 is 13.03%.

[0062] Preparation method two is as follows:

[0063] Weigh 2.75g of SBE-β-CD into a 100mL round-bottom flask, add 4mL of purified water, and dissolve by magnetic stirring in a 60℃ oil bath. Separately, accurately weigh 10mg of SN-011 into a centrifuge tube, add an appropriate amount of anhydrous ethanol and sonicate to dissolve, then slowly add to the SBE-β-CD solution. Heat and stir for 4 hours, then turn off the heat and continue stirring for 1 hour. After cooling the inclusion complex solution to room temperature, vacuum rotary evaporate it in a 37℃ water bath for 30 minutes to remove the solvent. Add 4mL of purified water and hydrate at 37℃ for 10 minutes. Transfer the solution to a 10mL volumetric flask, dilute with purified water to the mark, shake well, and filter through a 0.22μm filter membrane to remove insoluble matter.

[0064] Finished product testing: The encapsulation rate of SN-011 was tested and calculated according to preparation method one, and was 83.23%.

[0065] Preparation method three is as follows:

[0066] Weigh 2.75g of SBE-β-CD into a 100mL round-bottom flask, add 4mL of purified water, and dissolve by sonication. Separately, accurately weigh 10mg of SN-011 into a centrifuge tube, add an appropriate amount of anhydrous ethanol, dissolve by sonication, and slowly add it dropwise to the SBE-β-CD solution. Sonicate at 1500W for 4 hours, then remove and allow the inclusion complex solution to cool to room temperature. Then, vacuum rotary evaporate the solution in a 37℃ water bath for 30 minutes to remove the solvent. Add 4mL of purified water and hydrate at 37℃ for 10 minutes. Transfer the solution to a 10mL volumetric flask, dilute with purified water to the mark, shake well, and filter through a 0.22μm filter membrane to remove insoluble matter.

[0067] Finished product testing: The encapsulation rate of SN-011 was tested and calculated according to preparation method two, and was 42.02%.

[0068] The comparative experiments above show that the inclusion complex obtained by preparation method two has the best encapsulation rate.

[0069] Example 2

[0070] Prescription: SN-011 10mg

[0071] SBE-β-CD 1.871g

[0072] Add water to 10mL

[0073] Preparation method: Weigh 1.871 g of SBE-β-CD into a 100 mL round-bottom flask, add 4 mL of purified water, and dissolve by magnetic stirring in a 40 °C oil bath. Separately, accurately weigh 10 mg of SN-011 into a centrifuge tube, add 1.5 mL of anhydrous ethanol, sonicate to dissolve, and then slowly add to the SBE-β-CD solution, followed by 2.5 mL of anhydrous ethanol. Heat and stir for 4 h, then turn off the heat and continue stirring for 1 h. After cooling the inclusion complex solution to room temperature, vacuum rotary evaporate at a 37 °C water bath for 30 min to remove the solvent. Add 4 mL of purified water and hydrate at 37 °C for 10 min. Transfer the solution to a 10 mL volumetric flask, add purified water to the mark, shake well, and filter through a 0.22 μm polyethersulfone membrane. No insoluble particles / visible foreign matter were observed when placed at room temperature.

[0074] Finished product testing: The encapsulation rate of SN-011 was tested and calculated to be 89.73% according to the method in Example 1.

[0075] Example 3

[0076] Prescription: SN-011 10mg

[0077] SBE-β-CD 2.806g

[0078] Add water to 10mL

[0079] Preparation method: Weigh 2.806 g of SBE-β-CD into a 100 mL round-bottom flask, add 5.6 mL of purified water, and dissolve by magnetic stirring in a 40 °C oil bath. Separately, accurately weigh 10 mg of SN-011 into a centrifuge tube, add an appropriate amount of anhydrous ethanol and sonicate to dissolve, then slowly add the SN-β-CD solution dropwise, and add anhydrous ethanol to a final volume of 14 mL. Heat and stir for 4 h, then turn off the heat and continue stirring for 1 h. After cooling the inclusion complex solution to room temperature, vacuum rotary evaporate it in a 37 °C water bath for 30 min to remove the solvent. Add 4 mL of purified water and hydrate at 37 °C for 10 min. Transfer the solution to a 10 mL volumetric flask, add purified water to the mark, shake well, and filter through a 0.22 μm polyethersulfone membrane. No insoluble particles / visible foreign matter appeared after 30 days at room temperature.

[0080] Finished product testing: The encapsulation rate of SN-011 was tested and calculated to be 99.66% according to the method in Example 1.

[0081] Example 4

[0082] Prescription: SN-011 10mg

[0083] SBE-β-CD 2.75g

[0084] Povidone K12 15mg

[0085] Add water to 10mL

[0086] Preparation method: Weigh 2.75g of SBE-β-CD into a 100mL round-bottom flask, add 4mL of purified water, and dissolve by magnetic stirring in a 40℃ oil bath. Separately, accurately weigh 10mg of SN-011 into a centrifuge tube, add an appropriate amount of anhydrous ethanol and sonicate to dissolve, then slowly add to the SBE-β-CD solution, and add anhydrous ethanol to a final volume of 13.5mL. Heat and stir for 4h, then turn off the heat and continue stirring for 1h. After cooling the inclusion complex solution to room temperature, vacuum rotary evaporate in a 37℃ water bath for 30min to remove the solvent. Add 4mL of purified water and hydrate at 37℃ for 10min. Take out the hydrated solution, add 15mg of povidone K12, and magnetically stir for 30min at room temperature. Transfer the solution to a 10mL volumetric flask, add purified water to the mark, shake well, and filter through a 0.22μm polyethersulfone membrane. No insoluble particles / visible foreign matter appeared after standing at room temperature for 30 days.

[0087] Finished product testing: The encapsulation rate of SN-011 was tested and calculated to be 101.40% according to the method in Example 1.

[0088] Stability test

[0089] Take 5 mL of the sample solution prepared in this embodiment and place it in a 10 mL vial, sealing it with a cap. Place the vials in constant temperature chambers at 60, 40, and 30°C, a refrigerator at 4°C, and a light chamber with an irradiance of 4500±500 lx for 10 days each. Samples were taken on days 0, 5, and 10 of the experiment. The effects of different temperatures and strong light conditions on the sample solution were investigated, focusing on changes in appearance, visible foreign matter, and content. The results are shown in the table below:

[0090]

[0091] The above experiments show that the content of the formulation decreased slightly after being stored at 60℃ for 10 days, and the solution properties changed under 60℃ and strong light conditions. Under other conditions, the content did not decrease significantly after 10 days, remaining within the range of 95%–105% of the labeled amount, and other tested items showed no significant changes. This indicates that the SN-011 solubilization system is easily oxidized under high temperature and light conditions, exhibiting poor stability. Therefore, the storage temperature for the SN-011 solubilization system should be 4℃, and it should be protected from light.

[0092] A dilution study was conducted to assess the possibility of precipitation during dilution before administration to patients. The formulation prepared in this example was diluted 10-fold with physiological saline and 5% glucose injection, respectively. After standing at room temperature for 24 hours, the samples were filtered through a 0.22 μm polyethersulfone membrane. The filtrate was diluted 5-fold with methanol, and the SN-011 content was tested and calculated according to the method of Comparative Example 1. The results are shown in the table below:

[0093]

[0094] This experiment shows that it is possible to use povidone K12 as a stabilizer to prepare SN-011 injection that does not precipitate when diluted for delivery to patients. The choice of dilution medium has a certain impact on the content of SN-011. In clinical use, normal saline is preferred for infusion administration.

[0095] Example 5

[0096]

[0097] Preparation method: Weigh 2.338 g of SBE-β-CD into a 100 mL round-bottom flask, add 3.9 mL of purified water, and dissolve by magnetic stirring in a 40 °C oil bath. Separately, accurately weigh 10 mg of SN-011 into a centrifuge tube, add an appropriate amount of anhydrous ethanol and sonicate to dissolve, then slowly add to the SBE-β-CD solution, and add anhydrous ethanol to a final volume of 7.8 mL. Heat and stir for 4 h, then turn off the heat and continue stirring for 1 h. After cooling the inclusion complex solution to room temperature, vacuum rotary evaporate in a 37 °C water bath for 30 min to remove the solvent. Add 4 mL of purified water and hydrate at 37 °C for 10 min. Take out the hydrated solution, add 15 mg of povidone K12, and magnetically stir for 30 min at room temperature. Transfer the solution to a 10 mL volumetric flask, add purified water to the mark, shake well, and filter through a 0.22 μm polyethersulfone membrane. No insoluble particles / visible foreign matter appeared after standing at room temperature for 30 days.

[0098] Finished product testing: The encapsulation rate of SN-011 was tested and calculated to be 102.31% according to the method in Example 1.

[0099] Example 6

[0100]

[0101] Preparation method: Weigh 2.338 g of SBE-β-CD into a 100 mL round-bottom flask, add 4 mL of purified water, and dissolve by magnetic stirring in a 60 °C oil bath. Separately, accurately weigh 10 mg of SN-011 into a centrifuge tube, add 1.5 mL of anhydrous ethanol, sonicate to dissolve, and slowly add to the SBE-β-CD solution, then add 2.5 mL of anhydrous ethanol. Heat and stir for 2 h, then turn off the heat and continue stirring for 1 h. After cooling the inclusion complex solution to room temperature, vacuum rotary evaporate in a 37 °C water bath for 30 min to remove the solvent. Add 4 mL of purified water and hydrate at 37 °C for 10 min. Take out the hydrated solution, add 15 mg of povidone K12 and 20 mg of ascorbic acid, and magnetically stir at room temperature for 30 min. Transfer the solution to a 10 mL volumetric flask, add purified water to the mark, shake well, and filter through a 0.22 μm polyethersulfone membrane. No insoluble particles / visible foreign matter appeared after standing at room temperature.

[0102] Finished product testing: The encapsulation rate of SN-011 was tested and calculated to be 99.20% according to the method in Example 1.

[0103] Stability test

[0104] Take 5 mL of the sample solution prepared in this embodiment and place it in a 10 mL vial, sealing it with a cap. Place the vials in constant temperature chambers at 60, 40, and 30°C, a refrigerator at 4°C, and a light chamber with an irradiance of 4500±500 lx for 10 days each. Samples were taken on days 0, 5, and 10 of the experiment. The effects of different temperatures and strong light conditions on the sample solution were investigated, focusing on changes in appearance, visible foreign matter, and content. The results are shown in the table below:

[0105]

[0106] As can be seen from the above embodiments, the SN-011 solubilization system containing inclusion complex prepared by the formulation and preparation method of the present invention has a high encapsulation rate; the solubility of SN-011 in aqueous solution is significantly increased; and the SN-011 solubilization system has high stability under the conditions of 2 to 8℃.

[0107] Example 7

[0108]

[0109] Preparation method: Weigh 1.1g of HP-β-CD and place it in a suitable container. Add 1mL of PEG-400 and 5mL of purified water, and stir to dissolve in a 60℃ oil bath to obtain an HP-β-CD solution. Separately, accurately weigh 10mg of SN-011 and place it in a suitable container. Add 1mL of PEG-400, and stir to dissolve in a 60℃ oil bath to obtain an SN-011 solution. Add the HP-β-CD solution to the SN-011 solution, heat and stir for 2 hours, then turn off the heat and continue stirring for 1 hour to allow the inclusion complex solution to cool to room temperature. Transfer the obtained solution to a 10mL volumetric flask, add purified water to the mark, shake well, and filter through a 0.22μm polyethersulfone filter membrane to obtain the final product. No insoluble particles / visible foreign matter appeared when placed at room temperature, and no insoluble particles / visible foreign matter appeared after 10 days of observation at 2–8℃.

[0110] Finished product testing: The SN-011 content was tested and calculated to be 88.60% according to the method in Example 1.

[0111] Example 8

[0112]

[0113] Preparation method: Weigh 1.1g of HP-β-CD and place it in a suitable container. Add 2mL of PEG-400 and 5mL of purified water, and stir to dissolve in a 60℃ oil bath to obtain an HP-β-CD solution. Separately, accurately weigh 10mg of SN-011 and place it in a suitable container. Add 1mL of PEG-400, and stir to dissolve in a 60℃ oil bath to obtain an SN-011 solution. Add the HP-β-CD solution to the SN-011 solution, heat and stir for 2 hours, then turn off the heat and continue stirring for 1 hour to allow the inclusion complex solution to cool to room temperature. Transfer the obtained solution to a 10mL volumetric flask, add purified water to the mark, shake well, and filter through a 0.22μm polyethersulfone membrane to obtain the final product. No insoluble particles / visible foreign matter appeared when placed at room temperature, and no insoluble particles / visible foreign matter appeared after 10 days of observation at 2–8℃.

[0114] Finished product testing: The SN-011 content was tested and calculated to be 86.36% according to the method in Example 1.

[0115] Example 9

[0116]

[0117] Preparation method: Weigh 1.1g of HP-β-CD into a suitable container, add 2mL of PEG-400 and 5mL of purified water, and stir to dissolve in a 70℃ oil bath to obtain an HP-β-CD solution. Separately, accurately weigh 10mg of SN-011 into a suitable container, add 1mL of PEG-400, and stir to dissolve in a 70℃ oil bath to obtain an SN-011 solution. Add the HP-β-CD solution to the SN-011 solution, heat and stir for 2 hours, then turn off the heat and continue stirring for 1 hour to allow the inclusion complex solution to cool to room temperature. Transfer the obtained solution to a 10mL volumetric flask, add purified water to the mark, shake well, and filter through a 0.22μm polyethersulfone filter membrane to obtain the final product. No insoluble particles / visible foreign matter appeared when placed at room temperature, and no insoluble particles / visible foreign matter appeared after 10 days of observation at 2–8℃.

[0118] Finished product testing: The SN-011 content was tested and calculated to be 98.00% according to the method in Example 1.

[0119] Stability test

[0120] Take 5 mL of the sample solution prepared in this embodiment and place it in a 10 mL vial, sealing it with a cap. Place the vials in constant temperature chambers at 40℃ and 30℃, a refrigerator at 4℃, and a light chamber with an intensity of 4500±500 lx for 10 days each. Samples are taken on days 0, 5, and 10 of the experiment. The effects of different temperatures and strong light conditions on the sample solution are investigated, focusing on changes in appearance, visible foreign matter, and content. The results are shown in the table below:

[0121]

[0122] The above experiments show that the content of the formulation decreased after 10 days of exposure to light, and the solution properties changed. Under other conditions, the content did not decrease significantly after 10 days, and other tested items showed no significant changes. This indicates that the SN-011 solubilization system has high stability under normal storage conditions, but is easily oxidized and degraded under light.

[0123] Example 10

[0124]

[0125] Preparation method: Weigh 1.477 g of HP-β-CD and place it in a suitable container. Add 5 mL of purified water and dissolve in a 60°C oil bath by stirring to obtain an HP-β-CD solution. Separately, accurately weigh 10 mg of SN-011 and place it in a suitable container. Add 1 mL of PEG-400 and dissolve in a 60°C oil bath by stirring to obtain an SN-011 solution. Add the HP-β-CD solution to the SN-011 solution, heat and stir for 4 h, then turn off the heat and continue stirring for 1 h to allow the inclusion complex solution to cool to room temperature. Transfer the obtained solution to a 10 mL volumetric flask, add purified water to the mark, shake well, and filter through a 0.22 μm polyethersulfone membrane to obtain the final product. No insoluble particles / visible foreign matter appeared when placed at room temperature, and no insoluble particles / visible foreign matter appeared after 10 days of observation at 2–8°C.

[0126] Finished product testing: The SN-011 content was tested and calculated to be 88.70% according to the method in Example 1.

[0127] Example 11

[0128] 1. Establishment of a mouse model of psoriasis

[0129] The cGAS-STING pathway plays a crucial role in mammalian immunity. This pathway consists of cyclic GMP-AMP synthase (cGAS) and interferon gene stimulating factor (STING). cGAS recognizes exogenous DNA or leaked mitochondrial and genomic DNA, catalyzing the synthesis of cyclic GMP-AMP (cGAMP). cGAMP binds to and activates STING, initiating downstream signaling events that induce the production of interferon and other immune mediators, thereby activating innate immunity. Sustained activation of the cGAS-STING pathway has been shown to be associated with autoimmune diseases, and intervention in pathway dysregulation provides a potential avenue for targeted therapy.

[0130] Psoriasis is a common autoimmune skin disease, and the cGAS-STING pathway plays a key role in psoriatic inflammation. To evaluate the therapeutic potential of SN-011 cyclodextrin inclusion complex injection for skin inflammation, this study established a mouse model of psoriasis induced by imiquimod (IMQ) cream. Female Balb / C mice were selected as experimental animals and housed in an SPF-grade environment (constant temperature and a fixed 12-hour light-dark cycle), fed a standard diet, and given free access to water for one week of acclimatization. Before starting the experimental model, the backs of each mouse were shaved using a shaver. After removing most of the hair, an appropriate amount of depilatory cream was applied, and the area was wiped off after 1-2 minutes to form a 2×2 cm depilated area. The next day, except for the negative control group, 5% IMQ 62.5 mg was applied to the depilated area on the backs of the mice in the other groups for 8 consecutive days to establish the psoriasis model.

[0131] 2. Grouping and Dosing

[0132] Mice were randomly divided into 7 groups (n=5). Dexamethasone sodium phosphate was selected as the positive control drug because it has the highest anti-inflammatory efficacy among all corticosteroids and can inhibit the function of lymphocytes, macrophages, keratinocytes, and other immune cells. The sample solution was prepared as follows:

[0133] (1) SN-011 cyclodextrin inclusion complex injection: Weigh 2.75g of SBE-β-CD and place it in a 100mL round-bottom flask. Add 4mL of purified water and dissolve it by magnetic stirring in a 40℃ oil bath. Separately, accurately weigh 10mg of SN-011 and place it in a centrifuge tube. Add an appropriate amount of anhydrous ethanol and sonicate to dissolve it. Slowly add the solution to the SBE-β-CD solution and add anhydrous ethanol to a final volume of 13.5mL. Heat and stir for 4h, then turn off the heat and continue stirring for 1h to allow the inclusion complex solution to cool to room temperature. Then, vacuum rotary evaporate it in a 37℃ constant temperature water bath for 30min to remove the solvent. Add 4mL of 0.9% physiological saline and hydrate it at 37℃ for 10min. Take out the hydrated solution and add 15mg of povidone K12. Stir magnetically at room temperature for 30min and transfer the solution to a 10mL volumetric flask. Add physiological saline to the mark, shake well, and filter through a 0.22μm polyethersulfone filter membrane to obtain the final product.

[0134] (2) Blank excipient solution: Prepared by hydration with physiological saline without adding SN-011, following the preparation conditions described above.

[0135] (3) Dexamethasone sodium phosphate injection: Accurately weigh 10, 20 and 30 mg of dexamethasone sodium phosphate and place them in 10 mL volumetric flasks respectively. Add physiological saline to dissolve and dilute to the mark, shake well and let stand to obtain dexamethasone sodium phosphate solutions with concentrations of 1, 2 and 3 mg / mL.

[0136] Treatment procedure diagram (see) Figure 1 The specific drug administration information for each group of animals is shown in the table below:

[0137]

[0138] 2. Statistical Analysis

[0139] Statistical analysis was performed using GraphPad Prism software. The results of each experimental group are expressed as "Mean±SD (n=5)". One-way ANOVA was used for comparisons among multiple groups. P<0.05 was considered statistically significant.

[0140] 4. Morphological observation and PASI score

[0141] The PASI scoring method is used to assess the severity of psoriasis and the effectiveness of treatment in patients. It is currently the most widely used clinical indicator for quantitatively evaluating the skin lesions in psoriasis patients. The PASI scoring method includes four parts: lesion area, skin severity, symptoms and signs, and quality of life. The mouse PASI score mainly focuses on skin severity, including three indicators: skin thickness, scaling, and erythema. The scoring criteria for each indicator are shown in Table 2.

[0142] During the experiment, the severity of skin inflammation was recorded daily by photograph. Following the PASI scoring criteria, a 5-point scale (0-4) was used to score the inflammation, with the sum of the three scores serving as the evaluation indicator. The PASI scoring criteria for mice are shown in the table below:

[0143]

[0144] Figure 2 and Figure 3 Morphological images and PASI scores for scaling, erythema, and thickening in different animal groups were described. Daily morphological observations showed that mice in each group developed infiltrative erythema and skin thickening from day 2, with some showing scattered petechiae and other psoriasis-like changes, which worsened over time. By day 5, the skin had developed obvious thick, silvery-white scales with significant elevation, at which point the PASI score reached a peak of 7, indicating that the IMQ-induced psoriasis model had been successfully established. Treatment with appropriate drugs was then initiated. Treatment with different doses of SN-011 cyclodextrin inclusion complex injection and dexamethasone sodium phosphate significantly improved the skin lesions, almost returning to normal levels by the end of treatment. In contrast, the erythema on the backs of mice in the model group and the blank excipient group showed slight reduction, but the scaling and elevation did not significantly improve. This indicates that intervention with SN-011 cyclodextrin inclusion complex injection can inhibit the progression of psoriasis and has a certain therapeutic effect on psoriasis.

Claims

1. A pharmaceutical composition of SN-011, characterized by, The composition comprises SN-011, cyclodextrin and solvent, the concentration of SN-011 is 0.5-2 mg / mL, and the molar ratio of SN-011 to cyclodextrin is 1:(30-60). The cyclodextrin is selected from the group consisting of α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, sulfobutyl betadex sodium, hydroxypropyl betadex, and dimethyl betadex. The solvent is water or a mixed solvent of water and organic solvent.

2. The pharmaceutical composition of SN-011 according to claim 1, characterized in that, The organic solvent is selected from the group consisting of ethanol, propylene glycol, butylene glycol, isopropyl alcohol, glycerol, dimethylacetamide, polyethylene glycol 300, polyethylene glycol 400, and polyethylene glycol 600.

3. The pharmaceutical composition of SN-011 according to claim 2, characterized in that, The volume concentration of the organic solvent in the mixed solvent is 5-50%.

4. The pharmaceutical composition of SN-011 according to claim 1, characterized in that, The cyclodextrin is sulfobutyl betadex sodium or hydroxypropyl betadex.

5. The pharmaceutical composition of SN-011 according to claim 4, characterized in that, The pharmaceutical composition of SN-011 is selected from one of the following combinations: (1) comprising SN-011, sulfobutyl betadex sodium and water, the concentration of SN-011 is 0.5-2 mg / mL, and the molar ratio of SN-011 to sulfobutyl betadex sodium is 1:(30-60); (2) comprising SN-011, hydroxypropyl betadex and a mixed solvent of water and organic solvent, the concentration of SN-011 is 0.5-2 mg / mL, the molar ratio of SN-011 to hydroxypropyl betadex is 1:(30-60), and the volume concentration of the organic solvent in the mixed solvent is 5-50%.

6. The pharmaceutical composition of SN-011 according to claim 5, characterized in that, In combination (1), a stabilizer can also be added, the concentration of the stabilizer is 0.5-2.5 mg / mL, and the stabilizer is a water-soluble polymer or a pH regulator.

7. The pharmaceutical composition of any one of claims 1-6 for use in the preparation of a medicament for preventing or treating a disease mediated by STING signaling pathway.

8. Use according to claim 7, characterized in that, The medicament is any pharmaceutically acceptable dosage form prepared by using the pharmaceutical composition of any one of claims 1-6 as the main active ingredient and pharmaceutically acceptable pharmaceutical excipients.

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