Pharmaceutical composition for treating pulmonary fibrosis and preparation method thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a pharmaceutical composition for treating pulmonary fibrosis and a preparation method thereof. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 12 to 18 parts of radix astragali, 6 to 12 parts of radix ophiopogonis, 4 to 10 parts of poria cocos, 1 to 3 parts of honey-fried licorice root, 10 to 16 parts of radix salviae miltiorrhizae, 6 to 12 parts of radix scutellariae, 4 to 10 parts of cortex mori, 2 to 6 parts of pericarpium citri reticulatae, 1 to 4 parts of radix sophorae flavescentis and 1 to 4 parts of bulbus Three phases of polysaccharide, phenolic acid flavone and alkaloid are fractionated and compounded according to a certain ratio, and the polysaccharide is obtained through water extraction and alcohol precipitation and serves as a mucous membrane retention adhesion carrier; phenolic acid flavone is subjected to reflux extraction by using low-degree ethanol, so that the structural integrity is guaranteed and the stability is maintained; alkaloid is mildly salinized after being extracted with ethanol, stimulation is reduced, adsorption loss is reduced, polysaccharide is selectively lightly fermented to establish a slightly acidic environment, a buffer system is used for adjusting pH during compounding, cyclodextrin and other inclusion stable components are used, and three-phase synergistic intervention is performed on oxidation-inflammation-fibrosis.

Description

Technical Field

[0001] This invention belongs to the field of biopharmaceutical technology, specifically relating to a pharmaceutical composition for treating pulmonary fibrosis and its preparation method. Background Technology

[0002] Interstitial lung disease characterized by pulmonary fibrosis is characterized by insidious and irreversible progression. The core mechanisms involve persistent oxidative stress and amplified inflammation following epithelial injury, TGF-β and Smad-driven epithelial-mesenchymal transdifferentiation and fibroblast activation, and an imbalance between collagen deposition and matrix remodeling. First-line clinical treatment primarily uses pirfenidone and nintedanib, which can slow progression but are difficult to reverse scarring. Furthermore, adverse reactions and treatment costs limit long-term accessibility. Against this backdrop, traditional Chinese medicine (TCM), known for its multi-pathway regulation, is often used as an adjunct therapy. However, its prescriptions lack standardization and reproducibility, making it difficult to reliably cover the entire antioxidant-anti-inflammatory-antifibrotic pathway.

[0003] Existing traditional Chinese medicine preparations mostly employ single-method water decoction or single-method alcohol extraction. Hydrophilic polysaccharides and hydrophobic phenolic acids, flavonoids, and alkaloids interact and restrain each other in the same solvent system: polyphenols readily form non-specific associations with polysaccharides and proteins; flavonoids dissolve in their aglycone form but are difficult to absorb; and alkaloids, existing as free bases, exhibit poor dispersibility, strong bitterness and irritation, and undergo acid-base association and π-π stacking with polyphenols, leading to turbidity and sedimentation. While blind fermentation of the entire formula can improve taste, it often results in degradation of target components and increased batch-to-batch variability, making it difficult to obtain a stable slightly acidic environment and a proportion of absorbable structural components. In terms of dosage forms, decoctions and oral liquids are the main types, which suffer from slow early onset of action, poor clarity and shelf-life stability, and insufficient taste compliance. Quality control is mostly limited to the total polysaccharide and total flavonoid levels, lacking process markers (such as fermentation organic acids and post-fermentative agents) and structural markers (such as the ratio of free flavonoids to total flavonoids), making it difficult to support large-scale production. To address these pain points, some technical approaches have attempted advanced processes such as membrane separation, column chromatography purification, liposomes, and nanodelivery to improve bioavailability and stability. However, these processes involve significant equipment investment and process complexity, making them difficult to match with the existing production lines of most traditional Chinese medicine companies. While traditional Chinese medicine injections can increase exposure, they have extremely high requirements for aseptic assurance, pyrogen and allergy control, resulting in significant safety and compliance pressures. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the purpose of this invention is to provide a pharmaceutical composition for treating pulmonary fibrosis and a method for preparing the same.

[0005] The technical effects described in this invention are achieved through the following technical solution: a pharmaceutical composition for treating pulmonary fibrosis, comprising the following raw materials in parts by weight: 12-18 parts Astragalus membranaceus, 6-12 parts Ophiopogon japonicus, 6-10 parts Poria cocos, 1-3 parts prepared Glycyrrhiza uralensis, 10-16 parts Salvia miltiorrhiza, 6-12 parts Scutellaria baicalensis, 4-10 parts Morus alba root bark, 2-6 parts Citrus reticulata peel, 1-4 parts Sophora flavescens, and 1-4 parts Fritillaria cirrhosa.

[0006] Preferably, the pharmaceutical composition comprises the following raw materials in parts by weight: 15-18 parts Astragalus membranaceus, 8-12 parts Ophiopogon japonicus, 8-10 parts Poria cocos, 2-3 parts prepared Glycyrrhiza uralensis, 12-16 parts Salvia miltiorrhiza, 8-12 parts Scutellaria baicalensis, 6-10 parts Morus alba root bark, 3-6 parts Citrus reticulata peel, 2-4 parts Sophora flavescens and 2-4 parts Fritillaria cirrhosa. Preferably, the pharmaceutical composition comprises the following raw materials in parts by weight: 15 parts Astragalus membranaceus, 10 parts Ophiopogon japonicus, 8 parts Poria cocos, 2 parts prepared Glycyrrhiza uralensis, 14 parts Salvia miltiorrhiza, 10 parts Scutellaria baicalensis, 8 parts Morus alba root bark, 4 parts Citrus reticulata peel, 3 parts Sophora flavescens and 3 parts Fritillaria cirrhosa. Preferably, in any of the above-described pharmaceutical compositions for treating pulmonary fibrosis, the composition further comprises the following raw materials in parts by weight: 4 to 8 parts maltodextrin, 2 to 4 parts β-cyclodextrin, 1 to 2.5 parts glycerol, and 0.02 to 0.05 parts compound fermentation bacteria; Preferably, the compound fermentation bacteria consist of *Lactobacillus plantarum* and *Lactobacillus paracasei* in a 2:1 mass ratio; the activity of *Lactobacillus plantarum* is 1–5 × 10⁻⁶. 10 CFU / g; the activity of the *Lactobacillus paracasei* is 5–10 × 10⁻⁶ CFU / g. 10 CFU / g; Preferably, another aspect of the present invention provides a method for preparing a pharmaceutical composition for treating pulmonary fibrosis, comprising the following steps: S1: Add Astragalus membranaceus, Ophiopogon japonicus, Poria cocos and prepared licorice root to 10-12 times their weight of deionized water, extract at 90-100℃ for 60-90 min, filter, repeat the water extraction once, combine the two filtrates, concentrate at 60℃ to a relative density of 1.1-1.2 to obtain a concentrated solution; S2: Add 95% ethanol solution to the concentrate in step S1 to make the final volume fraction of the system 70-80%. After stirring, let it stand at room temperature for 8-12 hours, collect the precipitate, and dissolve it in deionized water at 60-65℃ to obtain the polysaccharide phase concentrate. S3: Take 60-70% of the polysaccharide phase concentrate from step S2 as the fermentation fraction, adjust the soluble solids to 8-12°Bx, inoculate with compound fermentation bacteria, statically ferment at 35-37℃ for 18-24 hours, treat at 72-75℃ for 15-20 minutes, and then cool to 35-40℃ to obtain the fermentation broth. S4: Reflux 50-60% ethanol for 60-90 min, repeat the reflux treatment once, combine the two filtrates, concentrate under reduced pressure to 20-30% solids, and remove alcohol until ethanol residue is <1% to obtain phenolic acid flavonoid phase concentrate. S5: Reflux Sophora flavescens and Fritillaria cirrhosa with 60-70% ethanol for 60-90 min, take the filtrate, slowly add citric acid to adjust the pH to 3.5-4, let stand for 30-60 min, concentrate under reduced pressure to 20-30% solids to obtain alkaloid phase concentrate. S6: Mix the fermentation broth from step S3 with the unfermented polysaccharide phase concentrate retained from step S2; then mix it with the phenolic acid flavonoid phase concentrate from step S4 and the alkaloid phase concentrate from step S5, adjust the pH to 5.5-6 with a buffer, add maltodextrin, β-cyclodextrin and glycerol in sequence, and stir at room temperature 200-400 rpm for 30-60 min to obtain the compound solution; S7: Degas the compound solution from step S6 under vacuum at -0.06 to -0.08 MPa for 10 to 20 minutes; then adjust the solid content to 28 to 32%, spray dry, collect the powder, and pass it through an 80-mesh sieve to obtain the pharmaceutical composition; Preferably, in step S4, the liquid-to-solid ratio of the ethanol reflux treatment is 8–10 mL / g; the reflux temperature is 78–82°C. Preferably, in step S5, the liquid-to-solid ratio of the ethanol reflux treatment is 6–10 mL / g; the reflux temperature is 75–80°C. Preferably, in step S6, the buffer is composed of citric acid and sodium citrate in a 1:1 molar ratio; Preferably, in step S7, the spray drying parameters are: feed temperature 35-45℃, inlet air temperature 145-158℃, outlet air temperature 72-80℃, and atomization pressure 0.2-0.35MPa.

[0007] The beneficial effects of this invention are as follows: Compared with existing technologies, this invention extracts and blends three phases—a polysaccharide phase for invigorating qi and antioxidation, a phenolic acid flavonoid phase for inhibiting fibrosis signals, and an alkaloid phase for inhibiting inflammation and fibroblast activation—in a fixed ratio to form a complex oral system carried by a hydrophilic matrix. First, a polysaccharide phase rich in arabinose and glucose residues is obtained through water extraction followed by hot alcohol precipitation, serving as a basic framework for mucosal retention and adhesion. Then, tanshinone and scutellaria baicalensis flavonoids are extracted by low-degree ethanol reflux to ensure the integrity of aromatic hydroxyl and carboxyl structures, facilitating dissociation equilibrium and chemical stability in a weakly acidic environment. Additionally, alkaloids from Sophora flavescens and Fritillaria cirrhosa are extracted with ethanol and gently salted, allowing them to exist in their salt form in the aqueous phase, reducing irritation and avoiding non-specific adsorption loss of polyphenols by alkaline components. During blending, the polysaccharide phase is continuous, while the phenolic acid flavonoids and salted alkaloids are uniformly embedded as dispersed phases. This allows carrier retention, pathway inhibition, and inflammation control to work synergistically within the same system, avoiding the mutual restraint of active components and unpleasant taste caused by traditional whole-formula decoction.

[0008] Based on phase-separation extraction, selective light fermentation and low-temperature inactivation are performed only on the polysaccharide phase. The lactic acid and metagenics produced during fermentation maintain the polysaccharide backbone while creating a slightly acidic environment. This enhances the chemical stability and transmembrane diffusion of phenolic acid flavonoids in the intestinal segment, and also forms a persistent hydration layer on the mucosal surface with extracellular polysaccharides, extending the local exposure time. This achieves a coupled amplification of carrier retention, slightly acidic stability, and passive diffusion. The fermentation window is limited to mild conditions, avoiding non-selective degradation of the phenolic acid backbone and alkaloid salts. At the same time, it provides a mild buffer for subsequent short-term acid-catalyzed deligation, allowing some flavonoid glycosides to convert to the free form, improving the penetration and first-pass loss of insoluble flavonoids, and compensating for the shortcomings of simple water-alcohol extraction in terms of oral absorption. In the compounding stage, a citric acid and sodium citrate buffer system stabilizes the pH of the formulation within a weakly acidic range, allowing phenolic acids to achieve a more balanced solubility and membrane permeability in an ionic-molecular coexistence form. Alkaloids remain in their salt form to reduce bitterness and local irritation. Small amounts of maltodextrin and cyclodextrin are added for inclusion complexation, inhibiting hydrophobic aggregation and π-π stacking between polyphenols and alkaloids, reducing heat-induced polymerization during spray drying or pasteurization. The polysaccharide and phenolic acid-flavonoid phases dominate the compounding ratio, providing a foundation for antioxidant and microcirculation improvement. The alkaloid phase, inserted at low doses, exerts targeted anti-inflammatory and activation-inhibiting effects, avoiding the taste and safety risks associated with high alkali loads. The polysaccharide phase improves the inflammatory microenvironment by enhancing mucosal adhesion and free radical scavenging, thus alleviating oxidative stress and epithelial damage. The phenolic acid and flavonoid phase, protected by a weak acid and carrier, maintains effective exposure, inhibiting TGF-β and Smad-mediated transcriptional activation and correcting epithelial-mesenchymal transdifferentiation, while simultaneously improving microvascular flow and matrix metalloproteinase imbalances and reducing collagen deposition tendency. The alkaloid phase, supported by salinization and inclusion, is stably released, inhibiting the inflammatory cascade and fibroblast α-SMA upregulation, synergistically weakening the abnormal expression of collagen I and fibronectin. This three-phase approach, with its carrier layer, chemical stabilization layer, and pathway intervention layer, achieves a stepwise synergy within the same drug delivery system. Compared to single decoction or fermentation systems, it more effectively achieves simultaneous control of the three pathological pathways of oxidation-inflammation-fibrosis, reducing the rebound risk associated with single-target interventions. Attached Figure Description

[0009] Figure 1 The cumulative dissolution results are for the pharmaceutical compositions prepared in Examples 1-3 and Comparative Examples 1-3 of this invention; Figure 2 The results show the absorbance changes of the pharmaceutical compositions prepared in Examples 1-3 and Comparative Examples 1-3 of this invention. Figure 3 The results show the α-SMA inhibition rates of the pharmaceutical compositions prepared in Examples 1-3 and Comparative Examples 1-3 of this invention. Figure 4 The results show the gel shrinkage rates of the pharmaceutical compositions prepared in Examples 1-3 and Comparative Examples 1-3 of this invention. Detailed Implementation

[0010] 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.

[0011] Example 1: A pharmaceutical composition for treating pulmonary fibrosis, comprising the following raw materials in parts by weight: 15 parts Astragalus membranaceus, 10 parts Ophiopogon japonicus, 8 parts Poria cocos, 2 parts prepared Glycyrrhiza uralensis, 14 parts Salvia miltiorrhiza, 10 parts Scutellaria baicalensis, 8 parts Morus alba root bark, 4 parts Citrus reticulata peel, 3 parts Sophora flavescens and 3 parts Fritillaria cirrhosa. The pharmaceutical composition further comprises the following raw materials in parts by weight: 6 parts maltodextrin, 3 parts β-cyclodextrin, 2 parts glycerol and 0.04 parts compound fermentation bacteria; The compound fermentation bacteria consist of *Lactobacillus plantarum* and *Lactobacillus paracasei* in a 2:1 mass ratio; the activity of *Lactobacillus plantarum* is 3 × 10⁻⁶. 10 CFU / g; the activity of the *Lactobacillus paracasei* was 8 × 10⁻⁶. 10 CFU / g; The preparation of the pharmaceutical composition for treating pulmonary fibrosis includes the following steps: S1: Add Astragalus membranaceus, Ophiopogon japonicus, Poria cocos and prepared licorice root to 11 times the weight of deionized water, extract at 98℃ for 75 min, filter, repeat the water extraction once, combine the two filtrates, concentrate at 60℃ to a relative density of 1.2 to obtain a concentrated solution. S2: Add 95% ethanol solution to the concentrate in step S1 to make the final volume fraction of the system 75%. After stirring, let it stand at room temperature for 10 hours, collect the precipitate, dissolve it in deionized water at 62°C, and obtain the polysaccharide phase concentrate. S3: Take 66% of the polysaccharide phase concentrate from step S2 as the fermentation fraction, adjust the soluble solids to 10°Bx, inoculate with compound fermentation bacteria, statically ferment at 36℃ for 20h, treat at 73℃ for 18min, and then cool to 37℃ to obtain the fermentation broth. S4: Salvia miltiorrhiza, Scutellaria baicalensis, Morus alba root bark and Citrus reticulata peel were refluxed with 55% ethanol for 80 min. The ethanol reflux treatment liquid-solid ratio was 9 mL / g and the reflux temperature was 80°C. The reflux treatment was repeated once. The two filtrates were combined and concentrated under reduced pressure to 25% solids. The alcohol was removed until the ethanol residue was <1% to obtain the phenolic acid flavonoid phase concentrate. S5: Sophora flavescens and Fritillaria cirrhosa were refluxed with 65% ethanol for 80 min. The ethanol reflux liquid-solid ratio was 8 mL / g and the reflux temperature was 78°C. The filtrate was collected, and citric acid was slowly added to adjust the pH to 3.7. The mixture was allowed to stand for 45 min and then concentrated under reduced pressure to 25% solids to obtain the alkaloid phase concentrate. S6: Mix the fermentation broth from step S3 with the unfermented polysaccharide phase concentrate retained from step S2; then mix it with the phenolic acid flavonoid phase concentrate from step S4 and the alkaloid phase concentrate from step S5. Adjust the pH to 5.7 using a buffer composed of citric acid and sodium citrate in a 1:1 molar ratio. Add maltodextrin, β-cyclodextrin and glycerol in sequence, and stir at 300 rpm for 50 min at room temperature to obtain the compound solution. S7: Degas the compound solution from step S6 under vacuum at -0.07 MPa for 15 min; then adjust the solid content to 30%, spray dry at a feed temperature of 40℃, an inlet air temperature of 152℃, an outlet air temperature of 75℃, and an atomization pressure of 0.28 MPa, collect the powder, and pass it through an 80-mesh sieve to obtain the pharmaceutical composition.

[0012] Example 2: A pharmaceutical composition for treating pulmonary fibrosis, comprising the following raw materials in parts by weight: 18 parts Astragalus membranaceus, 12 parts Ophiopogon japonicus, 10 parts Poria cocos, 3 parts prepared Glycyrrhiza uralensis, 16 parts Salvia miltiorrhiza, 12 parts Scutellaria baicalensis, 10 parts Morus alba root bark, 6 parts Citrus reticulata peel, 4 parts Sophora flavescens and 4 parts Fritillaria cirrhosa.

[0013] The pharmaceutical composition further comprises the following raw materials in parts by weight: 8 parts maltodextrin, 4 parts β-cyclodextrin, 2.5 parts glycerol and 0.05 parts compound fermentation bacteria; The compound fermentation bacteria consist of *Lactobacillus plantarum* and *Lactobacillus paracasei* in a 2:1 mass ratio; the activity of *Lactobacillus plantarum* is 5 × 10⁻⁶. 10 CFU / g; the activity of the *Lactobacillus paracasei* is 10 × 10⁻⁶. 10 CFU / g; The preparation of the pharmaceutical composition for treating pulmonary fibrosis includes the following steps: S1: Add Astragalus membranaceus, Ophiopogon japonicus, Poria cocos and prepared licorice root to 12 times the weight of deionized water, extract at 100℃ for 90 min, filter, repeat the water extraction once, combine the two filtrates, concentrate at 60℃ to a relative density of 1.1 to obtain a concentrated solution. S2: Add 95% ethanol solution to the concentrate in step S1 to make the final volume fraction of the system 80%. After stirring, let it stand at room temperature for 12 hours, collect the precipitate, dissolve it in deionized water at 65°C to obtain polysaccharide phase concentrate. S3: Take 60% of the polysaccharide phase concentrate from step S2 as the fermentation portion, adjust the soluble solids to 8°Bx, inoculate with compound fermentation bacteria, statically ferment at 35℃ for 18h, treat at 72℃ for 15min, and then cool to 35℃ to obtain the fermentation broth. S4: Salvia miltiorrhiza, Scutellaria baicalensis, Morus alba root bark and Citrus reticulata peel were refluxed with 50% ethanol for 60 min. The ethanol reflux treatment liquid-solid ratio was 8 mL / g and the reflux temperature was 78°C. The reflux treatment was repeated once. The two filtrates were combined and concentrated under reduced pressure to 20% solids. The ethanol was removed until the ethanol residue was <1% to obtain the phenolic acid flavonoid phase concentrate. S5: Sophora flavescens and Fritillaria cirrhosa were refluxed with 60% ethanol for 60 min. The ethanol reflux liquid-solid ratio was 6 mL / g and the reflux temperature was 75°C. The filtrate was collected, and citric acid was slowly added to adjust the pH to 3.5. The mixture was allowed to stand for 30 min and then concentrated under reduced pressure to 20% solids to obtain the alkaloid phase concentrate. S6: Mix the fermentation broth from step S3 with the unfermented polysaccharide phase concentrate retained from step S2; then mix it with the phenolic acid flavonoid phase concentrate from step S4 and the alkaloid phase concentrate from step S5. Adjust the pH to 5.5 with a buffer composed of citric acid and sodium citrate in a 1:1 molar ratio. Add maltodextrin, β-cyclodextrin and glycerol in sequence, and stir at 200 rpm for 30 min at room temperature to obtain the compound solution. S7: Degas the compound solution from step S6 under vacuum at -0.06 MPa for 10 min; then adjust the solid content to 28%, spray dry at a feed temperature of 35℃, an inlet air temperature of 145℃, an outlet air temperature of 80℃, and an atomization pressure of 0.2 MPa, collect the powder, and pass it through an 80-mesh sieve to obtain the pharmaceutical composition.

[0014] Example 3: A pharmaceutical composition for treating pulmonary fibrosis, comprising the following raw materials in parts by weight: 12 parts Astragalus membranaceus, 6 parts Ophiopogon japonicus, 6 parts Poria cocos, 1 part prepared Glycyrrhiza uralensis, 10 parts Salvia miltiorrhiza, 6 parts Scutellaria baicalensis, 4 parts Morus alba root bark, 2 parts Citrus reticulata peel, 1 part Sophora flavescens and 1 part Fritillaria cirrhosa.

[0015] The pharmaceutical composition further comprises the following raw materials in parts by weight: 4 parts maltodextrin, 2 parts β-cyclodextrin, 1 part glycerol and 0.02 parts compound fermentation bacteria; The compound fermentation bacteria consist of *Lactobacillus plantarum* and *Lactobacillus paracasei* in a 2:1 mass ratio; the activity of *Lactobacillus plantarum* is 1 × 10⁻⁶. 10 CFU / g; the activity of the *Lactobacillus paracasei* is 5 × 10⁻⁶. 10 CFU / g; The preparation of the pharmaceutical composition for treating pulmonary fibrosis includes the following steps: S1: Add Astragalus membranaceus, Ophiopogon japonicus, Poria cocos and prepared licorice root to 10 times their weight of deionized water, extract at 90℃ for 60 min, filter, repeat the water extraction once, combine the two filtrates, concentrate at 60℃ to a relative density of 1.1 to obtain a concentrated solution. S2: Add 95% ethanol solution to the concentrate in step S1 to make the final volume fraction of the system 70%. After stirring, let it stand at room temperature for 8 hours, collect the precipitate, and dissolve it with deionized water at 60°C to obtain the polysaccharide phase concentrate. S3: Take 70% of the polysaccharide phase concentrate from step S2 as the fermentation fraction, adjust the soluble solids to 12°Bx, inoculate with compound fermentation bacteria, statically ferment at 37°C for 24 hours, treat at 75°C for 20 minutes, and then cool to 40°C to obtain the fermentation broth. S4: Salvia miltiorrhiza, Scutellaria baicalensis, Morus alba root bark and Citrus reticulata peel were refluxed with 60% ethanol for 90 min. The ethanol reflux treatment liquid-solid ratio was 10 mL / g and the reflux temperature was 82°C. The reflux treatment was repeated once. The two filtrates were combined and concentrated under reduced pressure to 30% solids. The alcohol was removed until the ethanol residue was <1% to obtain the phenolic acid flavonoid phase concentrate. S5: Sophora flavescens and Fritillaria cirrhosa were refluxed with 70% ethanol for 90 min. The ethanol reflux liquid-solid ratio was 10 mL / g and the reflux temperature was 80°C. The filtrate was collected, and citric acid was slowly added to adjust the pH to 4. The mixture was allowed to stand for 60 min and then concentrated under reduced pressure to 30% of the solids to obtain the alkaloid phase concentrate. S6: Mix the fermentation broth from step S3 with the unfermented polysaccharide phase concentrate retained from step S2; then mix it with the phenolic acid flavonoid phase concentrate from step S4 and the alkaloid phase concentrate from step S5. Adjust the pH to 6 with a buffer composed of citric acid and sodium citrate in a 1:1 molar ratio. Add maltodextrin, β-cyclodextrin and glycerol in sequence, and stir at 400 rpm for 60 min at room temperature to obtain the compound solution. S7: Degas the compound solution from step S6 under vacuum at -0.08 MPa for 20 min; then adjust the solid content to 32%, spray dry, feed temperature 45℃, inlet air 158℃, outlet air 72℃, atomization pressure 0.35 MPa, collect the powder, and pass it through an 80-mesh sieve to obtain the pharmaceutical composition.

[0016] Comparative Example 1: The operation process of Comparative Example 1 is basically the same as that of Example 1. The main difference is that Comparative Example 1 omits the selective light fermentation in step S3 and directly uses the unfermented polysaccharide phase concentrate obtained in step S2 to participate in the compounding in step S6; the other operating parameters remain the same.

[0017] Comparative Example 2: The operation process of Comparative Example 2 is basically the same as that of Example 1. The main difference is that acidification and salting treatment is not performed in step S5 of Comparative Example 2, that is, citric acid is not used to adjust the pH, and the resulting alkaloid phase concentrate directly enters step S6 for compounding; the remaining operating parameters are the same.

[0018] Comparative Example 3: The operating procedures of Comparative Example 3 and Example 1 are basically the same. The main difference is that the formulation of Comparative Example 3 removes bitter herbs and fritillaria cirrhosa, does not prepare alkaloid phase concentrate, and uses polysaccharide phase to make up for the missing solids mass when compounding; the other operating parameters remain the same.

[0019] Performance testing: Physicochemical stability test: Take 0.5 g of each of the pharmaceutical compositions prepared in Examples 1-3 and Comparative Examples 1-3, add 100 mL of buffer medium (10 mM citric acid / sodium citrate, pH 5.8, 25℃), and stir magnetically until homogeneous; the addition time is defined as 0 min. At 0.5 min, 2 min, 5 min, 10 min, 20 min, and 30 min, take 2 mL of supernatant, filter through a 0.22 μm filter membrane, and immediately add an equal volume and equal composition of fresh medium; use HPLC (UV 286 nm) to quantify tanshinone B, calculate the cumulative dissolution amount (mg / g), and the results are as follows. Figure 1 As shown; Prepare 1% (w / v) solutions of the pharmaceutical compositions prepared in Examples 1-3 and Comparative Examples 1-3 respectively, using the same medium (10 mM citric acid and sodium citrate, pH 5.8, 25℃). After making up to volume and mixing well, let stand at 40℃. Define the moment of immersion in the solution as 0h. Take the supernatant at 0h and 24h respectively and place it in a 1cm cuvette. Measure the absorbance A600 at 600nm. Record the absorbance change ΔA = A600(24h) - A600(0h). The results are as follows. Figure 2 As shown.

[0020] based on Figure 1 and Figure 2Analysis of the results showed that the pharmaceutical compositions prepared in the embodiments of the present invention exhibited excellent overall dissolution effects with extremely low absorbance changes. However, the performance of Example 3 was significantly inferior to that of Examples 1 and 2. Based on the results of Comparative Example 1 and Example 1, Comparative Example 1 showed significantly delayed early and overall dissolution, with a faster increase in turbidity. This may be due to the lack of a lactic acid-dominated slightly acidic environment and a post-biotic matrix, resulting in a low ratio of free flavonoids to total flavonoids. This makes it easier for phenolic acid flavonoids and polysaccharides to form non-specific associations and aggregates, leading to a decrease in the equivalent diffusion coefficient. Simultaneously, the lack of extracellular polysaccharides produced during fermentation to support interfacial wetting and stable dispersion resulted in a reduced effective contact area and increased microagglomerations, manifested as a flat dissolution curve and increased absorbance changes. Based on the results of Comparative Example 2 and Example 1, Comparative Example 2 showed the slowest dissolution and the lowest absorbance. The largest increase in absorbance is due to the fact that alkaloids, in their free alkaloid form, readily aggregate and undergo acid-base association and π-π stacking with phenolic acids, forming hydrophobic microdomains and coarse micelles. This results in significant turbidity in the early stages, which occupies the active surface and inhibits the rapid release of phenolic acid flavonoids. Even if the final pH is uniformly set to weakly acidic, the heterogeneous aggregates formed in the early stages are difficult to completely reverse, thus impairing both stability and dissolution. Based on the results of Comparative Example 3 and Example 1, the dissolution and absorbance changes in Comparative Example 3 are at a moderate level, better than Comparative Examples 1 and 2 but lower than the Example. This may be because although the risk of aggregation caused by unsalted alkaloids is avoided, the hydrophilic microdomains and carrier dispersion effect brought about by the synergy of salted alkaloids and cyclodextrin are also lost. The compound system is inferior to the Example in terms of wetting, interfacial diffusion, and microscopic uniformity.

[0021] Cell function tests: Human embryonic lung fibroblasts MRC-5 were selected and divided into blank control, positive control (model control, TGF-β1 induced), Examples 1-3, and Comparative Examples 1-3. All samples were prepared with equal solids, equal drug volume, and final pH=5.5 to prepare a 10 mg / mL sterile aqueous solution, which was then diluted with DMEM working solution containing 1% FBS to a final concentration of 50 μg / mL (CCK-8 cell viability was tested, and cell viability at this concentration was >90%). MRC-5 cells were then injected at 1.5 × 10⁻⁶ cells / mL. 4Cells were seeded into 96-well plates and incubated overnight at 37°C with 5% CO2. The culture medium was discarded, and the samples were pretreated with fresh medium containing 1% FBS for 2 hours. The blank control group received no TGF-β1 or sample. The positive control group received TGF-β1 to 5 ng / mL. The treatment groups received TGF-β1 (5 ng / mL) and their respective samples (50 μg / mL). Incubation continued for 48 hours. After incubation, the cells were gently washed with PBS, and the lysis and dilution system provided by the ELISA kit was added. An α-SMA sandwich ELISA was performed according to the instructions. OD was read at 450 nm, and the concentration was calculated from the standard curve. The α-SMA inhibition rate was calculated as: (mean of positive control group - mean of each treatment group) / (mean of positive control group - mean of blank control group) × 100%. Each group had n=3 wells, and the results were independently replicated twice. Figure 3 As shown.

[0022] based on Figure 3 Results analysis showed that the pharmaceutical compositions prepared in the embodiments of the present invention generally exhibited excellent α-SMA inhibition rates. However, Example 3 showed the worst inhibition effect, which may be due to its significantly reduced ratio of alkaloids in the three-phase formulation, weaker microacid stability and interfacial dispersion, resulting in a lower inhibition amplitude, significantly inferior to Examples 1 and 2. Based on the results of Example 1 and Comparative Example 1, the α-SMA inhibition rate of Comparative Example 1 decreased significantly. This may be due to the lack of a microacidic environment and post-biotic matrix from fermentation, resulting in a lower ratio of free flavonoids and total flavonoids. Phenolic acid flavonoids and polysaccharides easily associate and aggregate, leading to a decrease in equivalent exposure and a significant decrease in α-SMA inhibition. Low; based on the results of Example 1 and Comparative Example 2, Comparative Example 2 showed the worst α-SMA inhibition effect. This may be because the alkaloids, in the form of free bases, undergo acid-base association and π-π stacking with phenolic acids, forming hydrophobic microclusters and increasing turbidity. The effective concentration of the active component and cell accessibility decreased synchronously, resulting in the worst functional endpoint. Based on the results of Example 1 and Comparative Example 3, the α-SMA inhibition of Comparative Example 3 was between that of Comparative Example 1 and Example 3. This may be because although the risk of unsalted aggregation was avoided, the targeted inhibition of fibroblast activation by alkaloids was lost, the three-phase coupling was weakened, and it was difficult to achieve the level of Example 3 by relying solely on polysaccharides and phenolic acid flavonoids.

[0023] Collagen gel shrinkage assay: Human lung fibroblasts MRC-5 were collected and divided into blank control, positive control (model control, TGF-β1 induced), Examples 1-3, and Comparative Examples 1-3. All samples were prepared with equal solids, equal drug volume, and final pH=5.5 to prepare a 10 mg / mL sterile aqueous solution, which was diluted to a final concentration of 50 μg / mL using DMEM working solution containing 1% FBS. Collagen gel was prepared in 24-well plates: Type I collagen mixture (final collagen 2 mg / mL, isotonic with 1×DMEM, pH adjusted to 7.4 with a small amount of NaOH) was prepared on ice, and MRC-5 suspension was added to a final concentration of 2×10⁻⁶.5 Cells / mL, dispensed 0.5 mL into each well, incubated at 37℃ for 30 min to gel; gently run a ring along the well wall to release the gel edge, add 0.5 mL of culture medium: blank control group without TGF-β1 and sample; positive control group with TGF-β1 to 5 ng / mL; treatment group with TGF-β1 (5 ng / mL) and corresponding sample (50 μg / mL), incubated at 37℃, 5% CO2, and the gel diameter (area) was recorded at 0 h and 48 h from above, and the area A0 (0 h) and A48 (48 h) were measured; the gel shrinkage rate (%) was calculated as (A0-A48) / A0×100%, and the results are as follows. Figure 4 As shown.

[0024] based on Figure 4 Results analysis showed that the pharmaceutical compositions prepared in the embodiments of the present invention generally exhibited superior gel shrinkage rates. However, the performance of Example 3 was significantly inferior to that of Examples 1 and 2. This may be due to the significant difference in its three-phase ratio compared to Examples 1 and 2, with a significantly reduced alkaloid content. Based on the results of Example 1 and Comparative Example 1, the gel shrinkage rate of Comparative Example 1 was higher than that of Example 1. This may be due to the lack of microacids and post-fermentative matrix from fermentation, resulting in a lower proportion of free and total flavonoids. Phenolic flavonoids are more likely to associate and aggregate with polysaccharides, leading to decreased equivalent exposure and cell accessibility, resulting in insufficient inhibition of the myofibroblast phenotype and an increased shrinkage rate. Analysis of the results of Example 1 and Comparative Example 2 showed that Comparative Example 2 had the worst performance. This may be because alkaloids in their free base form readily combine with phenolic acids to form acid-base associations and π-π stacking, resulting in hydrophobic microclusters and poor dispersion, leading to a double decrease in the effective concentration and accessibility of the active components. The three-phase synergy was disrupted, resulting in the lowest ability to inhibit stretching and stress fiber formation. Based on the analysis of the results of Example 1 and Comparative Example 3, the performance of Comparative Example 3 was between that of Comparative Example 1 and Example 2. This may be because although the risk of unsalted aggregation was avoided, the lack of alkaloids' directional inhibition of fibroblast activation and stretching signals weakened the three-phase coupling, making it difficult to fully inhibit contraction by polysaccharides and phenolic acid flavonoids alone.

[0025] 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 pharmaceutical composition for treating pulmonary fibrosis, characterized in that, Its composition includes the following raw materials in parts by weight: 12-18 parts Astragalus membranaceus, 6-12 parts Ophiopogon japonicus, 6-10 parts Poria cocos, 1-3 parts prepared Glycyrrhiza uralensis, 10-16 parts Salvia miltiorrhiza, 6-12 parts Scutellaria baicalensis, 4-10 parts Morus alba root bark, 2-6 parts Citrus reticulata peel, 1-4 parts Sophora flavescens and 1-4 parts Fritillaria cirrhosa.

2. The pharmaceutical composition for treating pulmonary fibrosis according to claim 1, characterized in that, The pharmaceutical composition comprises the following raw materials in parts by weight: 15 parts Astragalus membranaceus, 10 parts Ophiopogon japonicus, 8 parts Poria cocos, 2 parts prepared Glycyrrhiza uralensis, 14 parts Salvia miltiorrhiza, 10 parts Scutellaria baicalensis, 8 parts Morus alba root bark, 4 parts Citrus reticulata peel, 3 parts Sophora flavescens and 3 parts Fritillaria cirrhosa.

3. The pharmaceutical composition for treating pulmonary fibrosis according to claim 1, characterized in that, The pharmaceutical composition further comprises the following raw materials in parts by weight: 4-8 parts maltodextrin, 2-4 parts β-cyclodextrin, 1-2.5 parts glycerol, and 0.02-0.05 parts compound fermentation bacteria.

4. The pharmaceutical composition for treating pulmonary fibrosis according to claim 3, characterized in that, The compound fermentation bacteria consist of *Lactobacillus plantarum* and *Lactobacillus paracasei* in a 2:1 mass ratio; the activity of *Lactobacillus plantarum* is 1–5 × 10⁻⁶. 10 CFU / g; the activity of the *Lactobacillus paracasei* is 5–10 × 10⁻⁶ CFU / g. 10 CFU / g.

5. A method for preparing a pharmaceutical composition for treating pulmonary fibrosis according to any one of claims 1-4, characterized in that, Includes the following steps: S1: Add Astragalus membranaceus, Ophiopogon japonicus, Poria cocos and prepared licorice to deionized water, perform water extraction, filter, repeat water extraction, combine the two filtrates, concentrate to obtain concentrated solution; S2: Add ethanol solution to the concentrate from step S1, stir, let stand at room temperature, collect the precipitate, dissolve it in deionized water, and obtain polysaccharide phase concentrate. S3: Take a portion of the polysaccharide phase concentrate from step S2 as the fermentation fraction, adjust the soluble solids content, then inoculate with compound fermentation bacteria, perform static fermentation, heat sterilization, and then cool to obtain the fermentation broth. S4: Salvia miltiorrhiza, Scutellaria baicalensis, Morus alba root bark and Citrus reticulata peel were refluxed with ethanol, the reflux treatment was repeated, the two filtrates were combined, concentrated under reduced pressure, and treated with alcohol removal to obtain a concentrated phenolic acid flavonoid phase solution. S5: Sophora flavescens and Fritillaria cirrhosa were refluxed with ethanol, the filtrate was collected, citric acid was slowly added to adjust the pH, the mixture was allowed to stand, and the solution was concentrated under reduced pressure to obtain the alkaloid phase concentrate. S6: Mix the fermentation broth from step S3 with the unfermented polysaccharide concentrate retained from step S2. Then, mix the concentrated phenolic acid flavonoid phase solution from step S4 and the concentrated alkaloid phase solution from step S5 evenly, adjust the pH with a buffer, and add maltodextrin, β-cyclodextrin and glycerol in sequence. Stir at room temperature to obtain the compound solution. S7: Vacuum degas the compound solution from step S6; then adjust the solid content, spray dry, collect the powder, screen, and obtain the pharmaceutical composition.

6. A method for preparing a pharmaceutical composition for treating pulmonary fibrosis according to claim 5, characterized in that, In step S4, the liquid-to-solid ratio of the ethanol reflux treatment is 8-10 mL / g; the reflux temperature is 78-82°C.

7. A method for preparing a pharmaceutical composition for treating pulmonary fibrosis according to claim 6, characterized in that, In step S5, the liquid-to-solid ratio of the ethanol reflux treatment is 6–10 mL / g; the reflux temperature is 75–80°C.

8. A method for preparing a pharmaceutical composition for treating pulmonary fibrosis according to claim 7, characterized in that, In step S7, the spray drying parameters are: feed temperature 35-45℃, inlet air temperature 145-158℃, outlet air temperature 72-80℃, and atomization pressure 0.2-0.35MPa.

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