A combined pharmaceutical preparation for treating diabetic retinopathy and a method for preparing the same

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to a compound drug preparation for treating diabetic retinopathy and a preparation method thereof. The compound drug preparation is composed of traditional Chinese medicine raw materials and auxiliary materials. The traditional Chinese medicine raw materials comprise the following components: Astragalus extract, Danshen water-soluble extract, Danshen fat-soluble extract, Panax notoginseng total saponins, Huangqi extract, Ginkgo biloba leaf extract, Medlar polysaccharide extract and quercetin. The auxiliary materials comprise the following components: Soybean lecithin, Tween-80, PEG-40 hydrogenated castor oil, medium-chain triglyceride oil, glycerol, vitamin E, acacia gum, malt dextrin, microcrystalline cellulose, PVP K30, cross-linked sodium carboxymethyl cellulose and silicon dioxide. On the basis of conventional ophthalmic treatment, the application forms a closed-loop synergistic multi-target intervention retinal microenvironment through multiple components, solves the problems of component dispersion through a combination process, and takes into account biocompatibility and compliance.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a compound drug formulation for treating diabetic retinopathy and its preparation method. Background Technology

[0002] Diabetic retinopathy (DR) is a common and serious microvascular complication of diabetes. It is primarily caused by long-term hyperglycemia, leading to damage to retinal capillary endothelial cells and pericytes, resulting in increased vascular permeability, microaneurysms, punctate hemorrhages, and hard exudates. Further progression can lead to retinal ischemia, cotton wool spots, beaded vein changes, and intraretinal microvascular abnormalities (IRMA), even inducing neovascularization, causing vitreous hemorrhage, tractional retinal detachment, and other serious consequences, ultimately resulting in irreversible visual impairment or even blindness. Among the visual function declines associated with DR, diabetic macular edema (DME) has particularly important clinical significance: due to vascular leakage and fluid accumulation in the macular region, it causes changes in macular structure and leads to decreased vision. Clinically, optical coherence tomography (OCT) is often used to quantitatively assess retinal thickness, cystic edema, intraretinal / subretinal fluid, and vitreous-macular traction, thereby achieving objective monitoring of disease activity and treatment response.

[0003] Currently, the standard treatment system for DR / DME is relatively mature, mainly including intravitreal injection of anti-VEGF drugs, laser therapy, and surgical treatments such as vitrectomy. However, in real clinical practice, although the above standard treatments can effectively control disease progression and reduce the risk of blindness, a considerable proportion of patients still have the following unmet needs: First, some patients have unsatisfactory anatomical or visual function responses to anti-VEGF, manifested by residual leakage on OCT, insufficient retinal thickness reduction, or recurrent edema; Second, the occurrence and development of DR / DME are related to multiple factors such as chronic inflammation, oxidative stress, microcirculatory disorders, and neurovascular unit dysfunction, and single-targeted pathway therapy may not be able to cover all pathological aspects simultaneously; Third, anti-VEGF usually requires multiple, long-term repeated injections and frequent follow-ups, resulting in significant issues with treatment adherence, medical accessibility, and economic burden. In addition, in some pathways, the treatment strategy needs to be dynamically adjusted based on parameters such as visual acuity and retinal thickness, which objectively increases the overall treatment burden.

[0004] Based on the aforementioned clinical problems, an increasing number of studies are focusing on adjunctive therapy strategies that, without replacing standard treatment, improve the retinal microenvironment, reduce leakage and inflammatory response, and promote microcirculatory homeostasis through multi-target interventions. These strategies aim to enhance anatomical response, delay recurrence, and reduce the burden of treatment. Against this backdrop, compound traditional Chinese medicine preparations, due to their holistic regulatory characteristics and multi-component, multi-pathway effects, have been proposed as adjunctive therapy candidates for DR / DME. These preparations are intended to synergistically improve OCT structural indicators, visual function indicators, and imaging indicators reflecting the degree of leakage and ischemia, in addition to conventional anti-VEGF / laser therapy, thereby achieving the goal of improving treatment stability and reducing the need for relapse and retreatment. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the purpose of this invention is to provide a compound pharmaceutical preparation for treating diabetic retinopathy and its preparation method.

[0006] The technical effects described in this invention are achieved through the following technical solution: a compound drug preparation for treating diabetic retinopathy, which is composed of traditional Chinese medicine raw materials and excipients;

[0007] Preferably, the traditional Chinese medicine raw materials include the following components by weight: 12-20 parts of Astragalus membranaceus extract, 10-20 parts of Salvia miltiorrhiza water-soluble extract, 5-10 parts of Salvia miltiorrhiza fat-soluble extract, 8-15 parts of Panax notoginseng total saponins, 8-15 parts of Scutellaria baicalensis extract, 5-10 parts of Ginkgo biloba extract, 10-18 parts of Lycium barbarum polysaccharide extract, and 1-2 parts of quercetin;

[0008] Preferably, the tanshinone extract mainly contains water-soluble salvianolic acid extracts, wherein the total content of salvianolic acid components is ≥60wt%;

[0009] Preferably, the fat-soluble extract of *Salvia miltiorrhiza* mainly contains tanshinone concentrates, wherein the total content of tanshinone components is ≥40 wt%;

[0010] Preferably, the excipients comprise the following components by weight: 8-15 parts soybean lecithin, 3-8 parts Tween-80, 3-5 parts PEG-40 hydrogenated castor oil, 10-20 parts medium-chain triglyceride oil, 3-6 parts glycerol, 0.5-1.5 parts vitamin E, 10-20 parts gum arabic, 30-60 parts maltodextrin, 20-35 parts microcrystalline cellulose, 3-5 parts PVP K30, 2-5 parts croscarmellose sodium, and 0.5-1 part silicon dioxide;

[0011] Preferably, another aspect of the present invention provides a method for preparing a compound pharmaceutical formulation for treating diabetic retinopathy, specifically comprising the following steps:

[0012] S101: Add Astragalus extract, Salvia miltiorrhiza water-soluble extract, Scutellaria baicalensis extract, Lycium barbarum polysaccharide extract and quercetin to 6-10 parts by weight of purified water, then add 30-70% of the total amount of glycerol, stir at 600-900 rpm for 30-60 min to dissolve evenly, and obtain aqueous mother liquor.

[0013] S102: Dissolve the fat-soluble extract of Salvia miltiorrhiza in ethanol to a concentration of 2-5 wt%, then add 50-100% of the total vitamin E. Stir at 400-600 rpm for 10-30 min at 45-55℃ to obtain the organic phase. Take 50% of the total volume of the aqueous mother liquor from step S101, add 50-80% of the total volume of soybean lecithin and 50-80% of the total volume of Tween-80, and stir at 1000-1500 rpm for 20-40 min to obtain the stable aqueous phase. Slowly add the organic phase dropwise to the stable aqueous phase, perform high-speed shearing, and de-alcoholize under reduced pressure to obtain the nano-dispersion.

[0014] S103: Mix the nano-dispersion from step S102 with the remaining aqueous mother liquor from step S101, add total saponins of Panax notoginseng and Ginkgo extract, stir at 500-1000 rpm for 20-40 min, add the remaining parts by weight of soybean lecithin and Tween-80, and PEG-40 hydrogenated castor oil, stir and dissolve evenly to obtain the compound aqueous phase.

[0015] S104: Mix medium-chain triglyceride oil and the remaining weight of vitamin E, stir to dissolve evenly, and then slowly add it dropwise to the aqueous phase of the compound in step S103 within 10-20 minutes. High-speed shearing and high-pressure homogenization are then performed to obtain a fine emulsion.

[0016] S105: Add gum arabic and maltodextrin to the fine emulsion in step S104, and stir at 30-40℃ and 500-800 rpm for 30-60 min to obtain an emulsion; pre-cool the emulsion to 8-15℃, spray it with liquid nitrogen, freeze it to form frozen beads, collect it and freeze-dry it, then add microcrystalline cellulose, croscarmellose sodium and silica for wet preparation, dry at 60℃ until the water content is ≤5%, and pass it through a 12-24 mesh sieve to obtain a compound drug preparation;

[0017] Preferably, in step S102, the parameters of the high-speed shearing are: rotation speed 8000~14000rpm, time 6~12min, and system temperature controlled ≤35℃;

[0018] Preferably, in step S102, the parameters for vacuum deethanolination are: temperature 35–45°C, pressure -0.06–-0.09 MPa, and ethanol residue ≤0.5%;

[0019] Preferably, in step S104, the high-speed shearing parameters are: rotation speed 6000~12000rpm, time 5~15min, and system temperature controlled ≤35℃;

[0020] Preferably, in step S104, the high-pressure homogenization parameters are: pressure 50-100 MPa, number of cycles 3-6;

[0021] Preferably, in step S105, the parameters for injecting liquid nitrogen are: pressure nozzle 3-8 MPa, distance from nozzle to liquid nitrogen surface 10-25 cm, and freezing residence 4-8 min;

[0022] Preferably, in step S105, the freeze-drying parameters are: pre-freezing at -40 to -80°C for 2 to 4 hours; vacuum drying at -35 to -15°C and 50 to 80 Pa for 8 to 20 hours; and vacuum drying at 15 to 30°C and 30 to 50 Pa for 2 to 8 hours.

[0023] Preferably, in step S105, the wet formulation parameters are: main stirring 200-400 rpm, cutter 1200-2000 rpm, and spraying of binder for 5-10 min; the binder is obtained by adding the total amount of PVP K30 to purified water, stirring to dissolve evenly, and then adjusting the purified water to a concentration of 3-5 wt%.

[0024] The beneficial effects of this invention are as follows:

[0025] Compared with existing technologies, this invention, based on conventional anti-vascular endothelial growth factor therapy and related ophthalmic interventions, aims to reduce residual leakage and inflammation-driven relapse tendency by synergistically regulating the retinal microenvironment through multiple pathways, thereby potentially improving disease stability and reducing the burden of retreatment. Specifically, in terms of formulation, this invention is based on invigorating Qi and strengthening the body's resistance and improving microcirculation, combined with multiple active components that balance anti-inflammatory, antioxidant, and vascular homeostasis regulation, ensuring their effects are interconnected rather than simply additive. Astragalus and wolfberry components focus on enhancing the body's tolerance and repair foundation, providing support for vascular endothelial homeostasis and barrier function; Salvia miltiorrhiza water-soluble components and Scutellaria baicalensis flavonoids are more inclined to inhibit inflammatory responses and oxidative damage, reducing leakage-related microenvironmental stimulation; Salvia miltiorrhiza lipid-soluble components and vitamin E together form a lipid-phase protective chain, strengthening protection against membrane structure and lipid peroxidation; Panax notoginseng total saponins and Ginkgo biloba components complement each other in terms of blood perfusion and vascular function maintenance, helping to alleviate ischemia and endothelial stress caused by microcirculatory disorders. The aforementioned components form a closed-loop synergy of "barrier homeostasis - inflammation suppression - antioxidant protection - microcirculation support" in terms of efficacy focus, enabling the formulation to intervene in the pathological process at multiple targets in a relatively gentle and continuous manner, thus better suited to the positioning of adjuvant therapy and long-term management scenarios. In terms of process, this invention solves the common problems of uneven dispersion, phase separation and asynchronous release when lipid-soluble and water-soluble components coexist in compound formulations by using a combination of antisolvent nanodispersion and liquid nitrogen freeze-drying. The lipid-soluble extract achieves fine and stable dispersion through the antisolvent process, forming a stable interface with lecithin and surfactant system, making it easier for lipid-soluble activities to enter the subsequent emulsion system in a uniform form; subsequently, the oil phase is introduced to construct an oil-in-water emulsion droplet structure, allowing active components of different polarities to be synergistically carried within the same carrier framework. Liquid nitrogen freeze-drying rapidly shapes the droplet structure at low temperatures and forms a porous solid framework, which reduces the adverse effects of heat treatment on sensitive components and makes the formulation easier to redisperse and form a stable emulsion system when it is taken orally and comes into contact with water, thereby improving the consistency and reproducibility of the compound release and absorption process.

[0026] Furthermore, regarding stability and compatibility issues: First, the problem of easy oxidation and aggregation of lipid-soluble components is mitigated through oil-phase antioxidant protection and interface stabilization strategies, reducing activity loss and dispersion fluctuations during storage and use. Second, the potential viscosity and sedimentation risks of polysaccharides and flavonoids in the aqueous phase are improved through the combination of a solubilizing system and a solidified framework, making the system easier to process and more stable. Third, while emphasizing microcirculation improvement, the compound avoids an overly strong "single blood-activating" approach, instead focusing on barrier homeostasis, inflammation, and oxidative stress regulation, making it more suitable for routine ophthalmic treatment and adjunctive therapy in conjunction with basic medications for chronic diseases, thus achieving a balance between efficacy and safety. Fourth, the batch-to-batch variation and user experience issues common in traditional compound preparations are improved through a solid self-emulsifying structure and granular formulation, making the preparation more fluid and convenient for long-term management, and also helping to improve patient compliance. Attached Figure Description

[0027] Figure 1 This is a graph showing the permeability test results of the composite pharmaceutical formulations of Example 1 and Comparative Examples 1-4 of the present invention;

[0028] Figure 2 This is a graph showing the TNF-α concentration results of the anti-inflammatory test of the compound drug formulations of Example 1 and Comparative Examples 1-4 of this invention;

[0029] Figure 3 This is a graph showing the anti-inflammatory test results of the compound drug formulations of Example 1 and Comparative Examples 1-4 of the present invention. Detailed Implementation

[0030] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Unless otherwise specified, the raw materials involved in the present invention are all purchased through conventional commercial channels. Experimental methods without specific conditions are conventional methods and conditions well known in the art, or according to the conditions recommended by the instrument manufacturer.

[0031] Example 1: A compound pharmaceutical preparation for treating diabetic retinopathy, which is composed of traditional Chinese medicine raw materials and excipients;

[0032] The Chinese herbal raw materials include the following components by weight: 15 parts Astragalus membranaceus extract, 15 parts Salvia miltiorrhiza water-soluble extract, 8 parts Salvia miltiorrhiza fat-soluble extract, 12 parts total saponins of Panax notoginseng, 12 parts Scutellaria baicalensis extract, 8 parts Ginkgo biloba extract, 15 parts Lycium barbarum polysaccharide extract and 1.5 parts quercetin.

[0033] The excipients comprise the following components by weight: 12 parts soybean lecithin, 5 parts Tween-80, 4 parts PEG-40 hydrogenated castor oil, 15 parts medium-chain triglyceride oil, 5 parts glycerin, 1 part vitamin E, 15 parts gum arabic, 50 parts maltodextrin, 30 parts microcrystalline cellulose, 4 parts PVP K30, 3 parts croscarmellose sodium, and 0.8 parts silica.

[0034] The preparation of the compound drug formulation for treating diabetic retinopathy specifically includes the following steps:

[0035] S101: Add Astragalus extract, Salvia miltiorrhiza water-soluble extract, Scutellaria baicalensis extract, Lycium barbarum polysaccharide extract and quercetin to 8 times the weight of purified water, then add 50% of the total amount of glycerol, stir at 800 rpm for 45 min to dissolve evenly, and obtain aqueous mother liquor.

[0036] S102: Dissolve the fat-soluble extract of Salvia miltiorrhiza in ethanol to a concentration of 4 wt%, then add 80% of the total vitamin E. Stir at 500 rpm for 20 min at 50°C to obtain the organic phase. Take 50% of the total volume of the aqueous mother liquor from step S101, add 60% of the total volume of soybean lecithin and 60% of the total volume of Tween-80, and stir at 1200 rpm for 30 min to obtain the stable aqueous phase. Slowly add the organic phase dropwise to the stable aqueous phase at 10000 rpm for 10 min, controlling the system temperature to ≤35°C. Perform de-alcoholization under reduced pressure at 40°C and -0.08 MPa, with ethanol residue ≤0.5%, to obtain the nano-dispersion.

[0037] S103: Mix the nano-dispersion from step S102 with the remaining aqueous mother liquor from step S101, add total saponins of Panax notoginseng and Ginkgo extract, stir at 800 rpm for 30 min, add the remaining parts by weight of soybean lecithin and Tween-80, and PEG-40 hydrogenated castor oil, stir and dissolve evenly to obtain the compound aqueous phase.

[0038] S104: Mix medium-chain triglyceride oil and the remaining weight of vitamin E, stir and dissolve evenly, then slowly add it dropwise to the aqueous phase of the compound in step S103 over 15 min, with high-speed shearing at 10000 rpm for 10 min, controlling the system temperature ≤35℃; pressure 80 MPa, high-pressure homogenization 5 times to obtain a fine emulsion;

[0039] S105: Add gum arabic and maltodextrin to the fine emulsion from step S104, and stir at 35°C and 600 rpm for 50 min to obtain an emulsion; pre-cool the emulsion to 10°C, spray it into liquid nitrogen with a pressure nozzle of 5 MPa and a nozzle distance of 15 cm from the liquid nitrogen surface, freeze for 6 min to form frozen beads, collect them, pre-freeze at -60°C for 3 h; vacuum dry at -25°C and 60 Pa for 15 h; freeze dry at 25°C and 40 Pa for 5 h, then add microcrystalline cellulose, croscarmellose sodium, and silica for wet formulation, stir at 300 rpm, cut at 1600 rpm, spray with binder solution for 8 min, the binder solution is 4 wt% PVP K30 solution; dry at 60°C until the water content is ≤5%, and pass through a 12-24 mesh sieve to obtain a compound drug formulation.

[0040] Example 2: A compound pharmaceutical preparation for treating diabetic retinopathy, which is composed of traditional Chinese medicine raw materials and excipients;

[0041] The Chinese herbal raw materials include the following components by weight: 20 parts of Astragalus membranaceus extract, 20 parts of Salvia miltiorrhiza water-soluble extract, 10 parts of Salvia miltiorrhiza fat-soluble extract, 15 parts of Panax notoginseng total saponins, 15 parts of Scutellaria baicalensis extract, 10 parts of Ginkgo biloba extract, 18 parts of Lycium barbarum polysaccharide extract and 2 parts of quercetin.

[0042] The excipients comprise the following components by weight: 15 parts soybean lecithin, 8 parts Tween-80, 5 parts PEG-40 hydrogenated castor oil, 20 parts medium-chain triglyceride oil, 6 parts glycerin, 1.5 parts vitamin E, 20 parts gum arabic, 60 parts maltodextrin, 35 parts microcrystalline cellulose, 5 parts PVP K30, 5 parts croscarmellose sodium, and 1 part silicon dioxide.

[0043] The preparation of the compound drug formulation for treating diabetic retinopathy specifically includes the following steps:

[0044] S101: Add Astragalus extract, Salvia miltiorrhiza water-soluble extract, Scutellaria baicalensis extract, Lycium barbarum polysaccharide extract and quercetin to 10 times their weight of purified water, then add 70% of the total amount of glycerol, stir at 900 rpm for 30 min to dissolve evenly, and obtain aqueous mother liquor.

[0045] S102: Dissolve the fat-soluble extract of Salvia miltiorrhiza in ethanol to a concentration of 5 wt%, then add 100% of the total vitamin E. Stir at 600 rpm for 10 min at 55°C to obtain the organic phase. Take 50% of the total volume of the aqueous mother liquor from step S101, add 80% of the total volume of soybean lecithin and 80% of the total volume of Tween-80, and stir at 1500 rpm for 20 min to obtain the stable aqueous phase. Slowly add the organic phase dropwise to the stable aqueous phase at 14000 rpm for 6 min, controlling the system temperature to ≤35°C. Perform de-alcoholization under reduced pressure at 45°C and -0.09 MPa, with ethanol residue ≤0.5%, to obtain the nano-dispersion.

[0046] S103: Mix the nano-dispersion from step S102 with the remaining aqueous mother liquor from step S101, add total saponins of Panax notoginseng and Ginkgo extract, stir at 1000 rpm for 20 min, add the remaining parts by weight of soybean lecithin and Tween-80, and PEG-40 hydrogenated castor oil, stir and dissolve evenly to obtain the compound aqueous phase.

[0047] S104: Medium-chain triglyceride oil was slowly added dropwise to the aqueous phase of the compound from step S103 over 20 min, with high-speed shearing at 12000 rpm for 5 min, and the system temperature was controlled to be ≤35℃; the pressure was 100 MPa, and the high-pressure homogenization was performed 3 times to obtain a fine emulsion;

[0048] S105: Add gum arabic and maltodextrin to the fine emulsion from step S104, and stir at 800 rpm for 30 min at 40°C to obtain an emulsion; pre-cool the emulsion to 8°C, spray it into liquid nitrogen, pressurize the nozzle to 8 MPa, keep the nozzle 10 cm from the liquid nitrogen surface, freeze for 4 min to form frozen beads, collect them and pre-freeze at -80°C for 2 h; vacuum dry at -35°C, 50 Pa for 8 h; freeze dry at 30°C, 30 Pa for 2 h, then add microcrystalline cellulose, croscarmellose sodium, and silica for wet formulation, stir at 400 rpm, cut at 2000 rpm, spray the binder solution for 10 min, the binder solution is 5 wt% PVP K30 solution; dry at 60°C until the water content is ≤5%, and pass through a 12-24 mesh sieve to obtain the compound drug formulation.

[0049] Example 3: A compound pharmaceutical preparation for treating diabetic retinopathy, which is composed of traditional Chinese medicine raw materials and excipients;

[0050] The Chinese herbal raw materials include the following components by weight: 12 parts Astragalus membranaceus extract, 10 parts Salvia miltiorrhiza water-soluble extract, 5 parts Salvia miltiorrhiza fat-soluble extract, 8 parts total saponins of Panax notoginseng, 8 parts Scutellaria baicalensis extract, 5 parts Ginkgo biloba extract, 10 parts Lycium barbarum polysaccharide extract and 1 part quercetin.

[0051] The excipients comprise the following components by weight: 8 parts soybean lecithin, 3 parts Tween-80, 3 parts PEG-40 hydrogenated castor oil, 10 parts medium-chain triglyceride oil, 3 parts glycerin, 0.5 parts vitamin E, 10 parts gum arabic, 30 parts maltodextrin, 20 parts microcrystalline cellulose, 3 parts PVP K30, 2 parts croscarmellose sodium, and 0.5 parts silicon dioxide.

[0052] The preparation of the compound drug formulation for treating diabetic retinopathy specifically includes the following steps:

[0053] S101: Add Astragalus extract, Salvia miltiorrhiza water-soluble extract, Scutellaria baicalensis extract, Lycium barbarum polysaccharide extract and quercetin to 6 times the weight of purified water, then add 30% of the total amount of glycerol, stir at 600 rpm for 60 min to dissolve evenly, and obtain aqueous mother liquor.

[0054] S102: Dissolve the fat-soluble extract of Salvia miltiorrhiza in ethanol to a concentration of 2 wt%, then add 50% of the total vitamin E. Stir at 400 rpm for 30 min at 45°C to obtain the organic phase. Take 50% of the total volume of the aqueous mother liquor from step S101, add 50% of the total volume of soybean lecithin and 50% of the total volume of Tween-80, and stir at 1000 rpm for 40 min to obtain the stable aqueous phase. Slowly add the organic phase dropwise to the stable aqueous phase at 8000 rpm for 12 min, controlling the system temperature to ≤35°C. Perform de-alcoholization under reduced pressure at 35°C and -0.06 MPa, with ethanol residue ≤0.5%, to obtain the nano-dispersion.

[0055] S103: Mix the nano-dispersion from step S102 with the remaining aqueous mother liquor from step S101, add total saponins of Panax notoginseng and Ginkgo extract, stir at 500 rpm for 40 min, add the remaining parts by weight of soybean lecithin and Tween-80, and PEG-40 hydrogenated castor oil, stir and dissolve evenly to obtain the compound aqueous phase.

[0056] S104: Mix medium-chain triglyceride oil and the remaining weight of vitamin E, stir to dissolve evenly, and then slowly add it dropwise to the aqueous phase of the compound in step S103 over 10 min. Perform high-speed shearing at 6000 rpm for 15 min, controlling the system temperature to ≤35℃; perform high-pressure homogenization at 50 MPa for 6 times to obtain a fine emulsion.

[0057] S105: Add gum arabic and maltodextrin to the fine emulsion from step S104, and stir at 30°C and 500 rpm for 60 min to obtain an emulsion; pre-cool the emulsion to 15°C, spray it into liquid nitrogen with a pressure nozzle of 3 MPa and a nozzle distance of 10 cm from the liquid nitrogen surface, freeze for 8 min to form frozen beads, collect them, pre-freeze at -40°C for 4 h; vacuum dry at -35°C and 80 Pa for 20 h; freeze dry at 15°C and 50 Pa for 8 h, then add microcrystalline cellulose, croscarmellose sodium, and silica for wet formulation, stir at 200 rpm, cut at 1200 rpm, spray with binder solution for 5 min, the binder solution is 3 wt% PVP K30 solution; dry at 60°C until the water content is ≤5%, and pass through a 12-24 mesh sieve to obtain the compound drug formulation.

[0058] Comparative Example 1: The main difference between Comparative Example 1 and Example 1 is that the antisolvent nano-dispersion step is not performed in Comparative Example 1. Instead, the lipid-soluble extract of tanshinone is directly added to medium-chain triglyceride oil and vitamin E and stirred to dissolve before proceeding directly to the subsequent emulsification step. The remaining steps and parameters are consistent with those of Example 1.

[0059] Comparative Example 2: The main difference between Comparative Example 2 and Example 1 is that Comparative Example 2 does not use liquid nitrogen atomization freezing and freeze-drying solidification processes. Instead, after adding gum arabic and maltodextrin to the homogenized emulsion, it is dried using a conventional spray drying method with an inlet temperature of 140°C and an outlet temperature of 80°C to obtain solid powder. The remaining steps and parameters are consistent with those of Example 1.

[0060] Comparative Example 3: The main difference between Comparative Example 3 and Example 1 is that soybean lecithin is not added in Comparative Example 3, and is replaced with an equal mass of maltodextrin; the remaining steps and parameters are the same as in Example 1.

[0061] Comparative Example 4: The main difference between Comparative Example 4 and Example 1 is that no fat-soluble extract of tanshinone was added in Comparative Example 4, and it was replaced with an equal mass of maltodextrin; the remaining steps and parameters were the same as in Example 1.

[0062] Performance testing:

[0063] Reemulsification stability test: Take 1.0 g of the composite drug formulation particles prepared in Examples 1-3 and Comparative Examples 1-4, and weigh them after equilibration under the same temperature and humidity conditions. Add the weighed particles to 100 mL of a medium (purified water) preheated to 37°C. Place the container on a thermostatic magnetic stirrer and stir at 500 rpm, recording the timing point from sample addition. Observe the process from particle disintegration to the formation of a uniform emulsion dispersion. The endpoint is defined as the absence of obvious agglomeration, visible oil droplets, and uniform appearance observed by the naked eye. Record the time required to reach the endpoint as the reemulsification time. After the reemulsification endpoint is reached, immediately take the entire suspension for particle size characterization. Use dynamic light scattering to test the droplet size distribution at 25°C and record the D value. 90 The test results, as structural performance indicators, are shown in Table 1 below.

[0064] Table 1. Stability test results of reemulsified structures in the examples and comparative examples

[0065]

[0066] Table 1 shows the results for Examples 1-3 regarding reemulsification time and droplet size D. 90 The two indicators showed the best overall performance with minimal differences between them, indicating that the synergistic structural chain formed by nano-dispersion, the interfacial stabilization system, and liquid nitrogen freeze-drying solidification can function stably. Comparative Example 3 performed the worst; the lack of lecithin significantly reduced interfacial film-forming ability, making it difficult for droplets to form stably and prone to aggregation and oil floating, thus requiring a longer time to reach the endpoint of uniform appearance and D... 90 The significant increase in size demonstrates that the interfacial system is a key step in the self-emulsification structure reconstruction of solids. In Comparative Example 1, after eliminating the antisolvent nanodispersion, the lipophilic activity entered the oil phase and droplets in a coarser dispersion state, resulting in a significantly larger droplet size and wider distribution after reemulsification. This limited the reemulsification process and increased its time consumption, indicating that nano-refinement is crucial for obtaining a fine and uniform droplet structure. Although Comparative Example 2 retained the nanodispersion and interfacial system, the replacement of liquid nitrogen freeze-drying with spray drying resulted in the loss of the pore structure and freeze-drying advantages during solidification. The droplet reconstruction efficiency decreased during rehydration, manifested in the time and D... 90 All were inferior to the Examples but superior to Comparative Example 1. Comparative Example 4 only removed the fat-soluble extract; the remaining structural and process synergies remained, as did reemulsification and D. 90 It showed only moderate deterioration and no structural collapse.

[0067] Accelerated stability testing: Oral granules from Example 1 and Comparative Examples 1-4 were packaged into small packages (1.0g granules per package) in the same packaging format, sealed, and stored under accelerated conditions. Accelerated conditions were 40°C and 75% relative humidity; samples were protected from light. Samples were taken at 0, 2, 4, and 8 weeks. After each sampling, appearance and clumping were checked (for clumping or discoloration), followed by water content determination. The structural properties were then retested using the same re-emulsification process as in Experiment 1: 1.0g of the sample at that time point was added to 100 mL of purified water preheated to 37°C, and the mixture was magnetically stirred at 500 rpm until the system was homogeneous and free of visible agglomeration and floating oil. The re-emulsification time was recorded. Immediately after the endpoint, the emulsion was tested for particle size distribution, preferably using dynamic light scattering at 25°C, and the particle size distribution (D) was recorded. 90 The test results are shown in Tables 2 and 3 below.

[0068] Table 2. Results of accelerated stability tests and moisture content for the examples and comparative examples.

[0069]

[0070] Table 3. Reemulsification Results of Accelerated Stability Tests in Examples and Comparative Examples

[0071]

[0072] Based on the results in Tables 2 and 3, Example 1 showed virtually no significant clumping or discoloration in the accelerated stability test, with only a slight increase in water content. Meanwhile, the reemulsification time and D... 90 Only slight drift. Comparative Example 3 showed the most significant deterioration, with earlier and more pronounced clumping accompanied by a deepening of yellowing, and the largest increase in water content; correspondingly, the reemulsification time was rapidly prolonged and D... 90 Significantly increased; the lack of lecithin leads to insufficient interfacial film-forming ability, and the droplets rely more on non-specific surfactant adsorption during solidification and storage. Under hot and humid conditions, interfacial film relaxation, droplet fusion, and solid powder adhesion are more likely to occur, ultimately resulting in heavier clumps, slower rehydration, and coarser droplets. Comparative Example 2 can still reemulsify relatively quickly at 0 weeks, but with the extension of acceleration time, more obvious clumping and color deepening occur, accompanied by reemulsification time and D. 90 The concentration of precipitates continues to rise. After the solidification method was changed from liquid nitrogen freeze-drying to spray drying, the pore structure and freeze-setting advantages of the powder were lost, making it more prone to localized melting and adhesion, glass transition on the particle surface, and pore collapse under hot and humid conditions. This leads to a decrease in rehydration penetration and droplet reconstruction efficiency. Comparative Example 1 showed a significant drift. After removing the antisolvent nano-dispersion, the lipophilic active material entered the oil phase and droplet system in a relatively coarse dispersion state. During storage, hydrophobic phase rearrangement and aggregation were more likely to occur. Under hot and humid conditions, this aggregation was more easily amplified, resulting in a shift in the droplet size distribution towards the larger particle size side after rehydration. 90The increase was significant, and the reemulsification time was correspondingly prolonged. Comparative Example 4 was generally similar to the Example in appearance and moisture content, but the reemulsification time and D... 90 The system still exhibits moderate drift, reflecting that its structural carrier and interface system can still provide basic stability, while the upper limit of performance is limited by the absence of the lipophilic module. Without the lipophilic extract of Tanshinone, the hydrophobic activity of the oil phase is reduced, and the viscoelasticity of the oil phase and the density of the interface film of the system may be reduced. This makes it easier for the freeze-dried solid self-emulsifying structure to form slightly coarser droplets during rehydration reconstruction, and it is more likely to experience a slight decrease in reconstruction efficiency under thermal and humid stress. Therefore, it exhibits a larger drift than the example.

[0073] Permeability test: Human retinal microvascular endothelial cells (HRMEC) were used. Transwell chambers with a pore size of 0.4 μm were used to insert the cells at a density of 1 × 10⁻⁶ cells / mL. 5 1 / well was inoculated into the upper chamber, and cultured in serum-containing medium until a stable monolayer was formed, usually for 3 days with daily medium changes. Once the monolayer became dense, a damage model was established, using high glucose stimulation (using low glucose DMEM as a substrate and supplementing D-glucose to a final concentration of 30 mM) combined with one of the inflammatory / leakage stimulating factors (TNF-α) for 24 hours to induce an increase in barrier permeability. The test samples were prepared into a dosing solution according to the equivalent active components: oral granules (Examples 1 and Comparative Examples 1-4) were weighed to ensure that the total amount of the corresponding active components of the traditional Chinese medicine was 50 mg, added to 100 mL of sterile 37°C medium, stirred at 500 rpm for 5 min to fully re-emulsify and form a uniform dispersion system, and after standing for 10 min, the uniform suspension was taken as the stock solution; the supernatant was filtered through a 0.45 μm sterile filter membrane to remove bacteria and particulate matter, and diluted with medium to the working concentration (100 μg / mL based on the total active components). The working solution was added to the upper chamber for 6 hours of treatment. A normal control group (no model stimulation), a model control group (with stimulation but no sample added), and comparative groups were set up. The permeability index was preferably measured using FITC-dextran flux: at the end of treatment, FITC-dextran (40 kDa, final concentration 1.0 mg / mL) was added to the upper chamber. After incubation at 37℃ for 60 minutes, the liquid in the lower chamber was removed, and the fluorescence intensity was measured. The relative permeability (%) was calculated as follows: (fluorescence intensity of the experimental group / fluorescence intensity of the model control group) × 100% under the same time and volume conditions. Results are as follows: Figure 1 As shown.

[0074] based on Figure 1As a result, compared with the model control, Example 1 significantly reduced FITC-dextran flux, suggesting that it can effectively improve endothelial barrier permeability. Comparative Example 3 showed the smallest decrease in permeability after removing lecithin. The absence of lecithin significantly reduced interfacial film formation and droplet stability, making droplet fusion, uneven dispersion, or loss of adsorption of effective components more likely after rehydration, resulting in insufficient and more volatile effective exposure for cells, making it difficult to form a sustained barrier protection effect. In Comparative Example 1, the removal of nano-dispersion made it easier for lipid-soluble components to exist in a coarser dispersion state. Even with lecithin and surfactant systems, it was more difficult to form small, stable droplets, leading to lower effective exposure and greater susceptibility to filtration or sedimentation. While Comparative Example 2 retained nano-dispersion and the interfacial system, spray drying solidification reduced the porous structure and low-temperature setting advantages. The rehydration reconstruction speed and post-reconstruction droplet stability were inferior to the freeze-dried system, resulting in less permeability improvement than the examples. Although Comparative Example 4 lacked the lipophilic module of Tanshinone, its interface system and structured carrier remained intact. After rehydration, the dispersion was more uniform and the exposure was more stable, thus it could still significantly reduce permeability. At the same time, the lack of the lipophilic module weakened the buffering capacity against membrane lipid peroxidation and membrane structural stress, resulting in a decrease in the degree of barrier improvement compared to the Example.

[0075] Anti-inflammatory test: Mouse macrophage line RAW264.7 was seeded in DMEM complete medium containing 10% fetal bovine serum and cultured at 37°C with 5% CO2 until the logarithmic growth phase. Cells were then collected and the cell density was adjusted to approximately 1×10⁻⁶. 5 Cells were seeded at a rate of 200 μL per well in 96-well plates and cultured overnight until the cells adhered stably. 100 mg of the compound drug formulations prepared in Example 1 and Comparative Examples 1-4 were added to 10 mL of DMEM medium containing 1% fetal bovine serum and incubated at 37°C for 4 h on a shaker at 120 rpm to extract the active components. The extracts were then centrifuged to remove insoluble particles and filtered through a 0.45 μm sterile membrane to obtain a sterile, clear extract for use in inflammatory cell experiments. The safe dilution factor for each extract without affecting cell viability was determined using the CCK-8 assay in preliminary experiments. The blank control group received no LPS or sample, while the model control group received LPS to a final concentration of 1 μg. Without adding samples, LPS (final concentration 1 μg / mL) and corresponding working solution (final concentration 100 μg / mL) were added to each treatment group simultaneously. Three parallel wells were set for each group. After incubation at 37℃ and 5% CO2 for 24 hours, the supernatant from each well was collected. The supernatant from each group was added to an ELISA plate pre-coated with TNF-α capture antibody. Incubation, washing, and color development were performed according to the kit instructions. The absorbance at 450 nm was measured, and the concentration of TNF-α in the supernatant of each group was calculated using a standard curve. The anti-inflammatory effect (%) was calculated as follows: (average TNF-α concentration in the model control group - average TNF-α concentration in each treatment group) / average TNF-α concentration in the model control group × 100%. The results are shown below. Figure 2 and Figure 3 As shown.

[0076] based on Figure 2 and Figure 3 As a result, Example 1 showed the most significant inhibitory effect on TNF-α secretion under LPS stimulation, suggesting that its compound anti-inflammatory pathway synergy was more complete and the release of effective components was more stable. Comparative Example 3 had the lowest inhibition rate, not simply due to the absence of an excipient, but because the lack of an interfacial stabilizing system made it more difficult for lipophilic and some hydrophobic anti-inflammatory components to exist in a stable and accessible state in the extract. Furthermore, these components were more prone to adsorption loss or phase separation during filtration, settling, and incubation, leading to a decrease in the proportion of effective components accessible to cells, ultimately resulting in insufficient TNF-α inhibition. The lack of nano-dispersion in Comparative Example 1 reduced the accessibility and uniformity of lipophilic components, weakening the anti-inflammatory effect. Although the change in the structural solidification method in Comparative Example 2 was less sensitive to the anti-inflammatory effect of the extract than to the reemulsification structure, the thermal stress and differences in the solid skeleton caused by spray drying may have led to a decrease in the activity of some sensitive components or a reduction in redispersion efficiency, thus resulting in a lower inhibition rate than the examples. In Comparative Example 4, the lack of lipophilic extract of tanshinone weakened the lipid-phase antioxidant / membrane protection chain, thus reducing the upper limit of inhibition during the inflammatory amplification process. However, its water-soluble anti-inflammatory components (scutellaria baicalensis flavonoids, tanshinone water-soluble components, quercetin, and astragalus / goji-related components) could still exert the main anti-inflammatory effects, thus still significantly reducing TNF-α. Overall, it performed better than Comparative Examples 1-3 but weaker than Example 1.

[0077] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A compound pharmaceutical preparation for treating diabetic retinopathy, characterized in that, It is composed of Chinese medicinal materials and excipients; the Chinese medicinal materials include the following components by weight: 12-20 parts of Astragalus membranaceus extract, 10-20 parts of Salvia miltiorrhiza water-soluble extract, 5-10 parts of Salvia miltiorrhiza fat-soluble extract, 8-15 parts of Panax notoginseng total saponins, 8-15 parts of Scutellaria baicalensis extract, 5-10 parts of Ginkgo biloba extract, 10-18 parts of Lycium barbarum polysaccharide extract and 1-2 parts of quercetin; The excipients comprise the following components by weight: 8-15 parts soybean lecithin, 3-8 parts Tween-80, 3-5 parts PEG-40 hydrogenated castor oil, 10-20 parts medium-chain triglyceride oil, 3-6 parts glycerol, 0.5-1.5 parts vitamin E, 10-20 parts gum arabic, 30-60 parts maltodextrin, 20-35 parts microcrystalline cellulose, 3-5 parts PVP K30, 2-5 parts croscarmellose sodium, and 0.5-1 part silicon dioxide. The water-soluble extract of *Salvia miltiorrhiza* mainly contains water-soluble salvianolic acid extracts, wherein the total content of salvianolic acid components is ≥60wt%; the fat-soluble extract of *Salvia miltiorrhiza* mainly contains tanshinone concentrates, wherein the total content of tanshinone components is ≥40wt%. The preparation method of the compound drug formulation specifically includes the following steps: S101: Add Astragalus extract, Salvia miltiorrhiza water-soluble extract, Scutellaria baicalensis extract, Lycium barbarum polysaccharide extract and quercetin to purified water, then add glycerin, stir to dissolve evenly, and obtain aqueous mother liquor; S102: Dissolve the fat-soluble extract of Salvia miltiorrhiza in ethanol, then add 50-100% of the total amount of vitamin E, stir and dissolve evenly to obtain an organic phase; add 50-80% of the total amount of soybean lecithin and 50-80% of the total amount of Tween-80 to the aqueous mother liquor of step S101, stir and disperse to obtain a stable aqueous phase; slowly add the organic phase dropwise to the stable aqueous phase, perform high-speed shearing and de-alcoholization under reduced pressure to obtain a nano-dispersion; S103: Mix the nano-dispersion from step S102 with the remaining aqueous mother liquor from step S101, add total saponins of Panax notoginseng and Ginkgo biloba extract, stir, add the remaining parts by weight of soybean lecithin and Tween-80, and PEG-40 hydrogenated castor oil, stir and dissolve evenly to obtain the compound aqueous phase. S104: Mix medium-chain triglyceride oil and the remaining weight of vitamin E, stir to dissolve evenly, and then slowly add it dropwise to the aqueous phase of the compound in step S103. High-speed shearing and high-pressure homogenization are performed to obtain a fine emulsion. S105: Add gum arabic and maltodextrin to the fine emulsion in step S104, stir and disperse evenly to obtain an emulsion; pre-cool the emulsion, spray it into liquid nitrogen, freeze it to form frozen beads, collect it and freeze dry it, then add microcrystalline cellulose, croscarmellose sodium and silica for wet preparation, dry it, and sieve it to obtain a compound drug preparation. In step S105, the wet formulation parameters are: main stirring 200-400 rpm, cutter 1200-2000 rpm, and spraying of binder for 5-10 min; the binder is obtained by adding the total amount of PVP K30 to purified water, stirring to dissolve evenly, and then adjusting the purified water to a concentration of 3-5 wt%.

2. The compound pharmaceutical preparation for treating diabetic retinopathy according to claim 1, characterized in that, In step S102, the parameters for high-speed shearing are: rotation speed 8000~14000rpm, time 6~12min, and system temperature ≤35℃; the parameters for depressurized alcohol removal are: temperature 35~45°C, pressure -0.06~-0.09MPa, and ethanol residue ≤0.5%.

3. The compound pharmaceutical preparation for treating diabetic retinopathy according to claim 1, characterized in that, In step S104, the high-speed shearing parameters are: rotation speed 6000~12000rpm, time 5~15min, and system temperature ≤35℃.

4. The compound pharmaceutical preparation for treating diabetic retinopathy according to claim 1, characterized in that, In step S104, the high-pressure homogenization parameters are: pressure 50-100 MPa, number of cycles 3-6.

5. A compound pharmaceutical preparation for treating diabetic retinopathy according to claim 1, characterized in that, In step S105, the parameters for injecting liquid nitrogen are: pressure nozzle 3-8 MPa, distance from nozzle to liquid nitrogen surface 10-25 cm, and freezing residence time 4-8 min.

6. The compound pharmaceutical preparation for treating diabetic retinopathy according to claim 1, characterized in that, In step S105, the freeze-drying parameters are: pre-freezing at -40 to -80°C for 2 to 4 hours; vacuum drying at -35 to -15°C and 50 to 80 Pa for 8 to 20 hours; and vacuum drying at 15 to 30°C and 30 to 50 Pa for 2 to 8 hours.

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