Polymorphic form and amorphous form of ziyuglycoside II sodium salt as well as preparation method and application of polymorphic form and amorphous form of ziyuglycoside II sodium salt

Abstract

The invention discloses a polymorphic form and an amorphous form of ziyuglycoside II sodium salt as well as a preparation method and application of the polymorphic form and the amorphous form, and particularly discloses physicochemical properties of the polymorphic form and the amorphous form of the ziyuglycoside II sodium salt (ZYG-II-Na) and application of the ziyuglycoside II sodium salt in preparation of medicines. Compared with ziyuglycoside II, the polymorphic ZYG-II-Na and the amorphous ZYG-II-Na obtained by means of crystal form screening and solid engineering have higher solubility and dissolution rate, and the bioavailability of the ziyuglycoside II in vivo is remarkably improved. In addition, in-vitro evaluation and pharmacokinetic tests show that the ZYG-II-Na has good pharmacokinetic characteristics and treatment potential when being used for treating lung diseases (such as through an inhalation way).

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to polymorphic and amorphous forms of Sanguisorba officinalis saponin II sodium salt, as well as preparation methods and applications thereof. Background Art

[0002] Chinese herbal medicine Sanguisorba officinalis ( Sanguisorba officinalis L. ) is derived from a plant in the Rosaceae family and is a traditional Chinese medicinal material. Its dried roots are widely used clinically to treat gastrointestinal bleeding and burn wound repair, and have demonstrated unique advantages in adjuvant cancer treatment. The main active ingredients of Sanguisorba officinalis are triterpenoid saponins, of which Ziyuglycoside II (ZYG II) is one of its core pharmacologically active ingredients. Studies have shown that ZYG II has multiple pharmacological activities, such as inducing cancer cell apoptosis and autophagy, inhibiting cell proliferation, inhibiting angiogenesis, and regulating the expression of inflammatory factors. These properties make it promising for the treatment of solid tumors such as gastric cancer, colorectal cancer, and lung adenocarcinoma. In addition, ZYG II also has potential therapeutic value in diseases such as type 2 diabetes and inflammation.

[0003] As a saponin drug, ZYG II suffers from poor water solubility and low bioavailability, which seriously impact its clinical efficacy. After oral administration of a 5 mg / kg dose of ZYG II to Sprague-Dawley (SD) rats, its absolute bioavailability was only 4.6%. Currently, approaches to improving the bioavailability of Sanguisorba officinalis active ingredients primarily focus on drug delivery systems, such as using self-microemulsifying systems or long-circulating liposome encapsulation to enhance their solubility and stability. However, it is worth noting that research specifically targeting ZYG II's own solubility and bioavailability remains elusive. Due to the extremely poor water solubility of ZYG II, the present invention selected its more water-soluble sodium salt (Sanguisorba officinalis saponin II sodium salt, ZYG-II-Na) and conducted screening of solid forms, including polymorphic and amorphous forms, to improve its solubility and optimize its pharmacokinetic behavior. Summary of the Invention

[0004] The purpose of the present invention is to provide the polymorphic form and amorphous form of Sanguisorba officinalis saponin II sodium salt and their preparation methods and applications, and specifically to provide the polymorphic form and amorphous form of Sanguisorba officinalis saponin II sodium salt (ZYG-Ⅱ-Na) to provide technical support for subsequent drug research and development and industrial production; the present invention also provides the preparation methods and applications of the polymorphic form and amorphous form of ZYG-Ⅱ-Na.

[0005] The polymorphic and amorphous forms of ZYG-Ⅱ-Na described in the present invention are specifically the polymorphic and amorphous forms of Sanguisorba officinalis saponin II sodium salt (referred to as Compound I). The structural formula of Compound I is as follows: .

[0006] The technical solutions of the present invention are as follows: The solid forms of Compound I include crystals (Form I, Form II, Form III, Form IV, Form V) and amorphous forms (Amorphous Form I, Amorphous Form II).

[0007] The X-ray powder diffraction spectrum of Form I expressed at a diffraction angle of 2θ±0.2° has diffraction peaks at 6.12°, 6.89°, 10.61°, 12.17°, 12.85°, 13.30°, 14.39°, 14.57°, 15.48°, 15.89°, 16.98°, 18.26°, and 22.63°.

[0008] Preferably, the X-ray powder diffraction spectrum of Form I expressed at a diffraction angle of 2θ±0.2° has diffraction peaks at 6.12°, 6.89°, 10.61°, 12.17°, 12.85°, 13.30°, 14.39°, 14.57°, 15.48°, 15.89°, 16.55°, 16.98°, 18.26°, 19.18°, and 22.63°.

[0009] More preferably, the X-ray powder diffraction spectrum of Form I expressed at a diffraction angle of 2θ±0.2° is 6.12°, 6.89°, 10.05°, 10.61°, 11.46°, 12.17°, 12.85°, 13.30°, 14.39°, 14.57°, 14.96°, 15.48°, 15.89°, 16.55°, 16.98°, 18.26°, 18.80°, 19.18°, 19.47°, 19.92°, 20.13°, 20.39°, 20.81°, There are diffraction peaks at 21.79°, 22.63°, 23.21°, 23.64°, 23.95°, 24.39°, 25.81°, 26.11°, 26.71°, 26.93°, 27.32°, 27.95°, 28.43°, 28.76°, 29.34°, 30.53°, 31.13°, 31.54°, 32.01°, 32.77°, 33.19°, 34.37°, 35.01°, 36.78°, 37.75°, and 39.06°.

[0010] Preferably, Form I has one or more of the following characteristics: The DSC curve of Form I has an endothermic peak at 130.6±3°C; The DSC curve of Form I has an endothermic peak at 244.5 ± 3 °C; The TGA curve of Form I has a decomposition endothermic peak starting point at 345.4±3°C.

[0011] The X-ray powder diffraction spectrum of Form II expressed at a diffraction angle of 2θ±0.2° has diffraction peaks at 5.75°, 7.09°, 9.74°, 11.53°, 13.28°, 14.08°, 15.03°, 15.40°, 18.21°, 18.46°, and 21.13°.

[0012] Preferably, the X-ray powder diffraction spectrum of Form II expressed at a diffraction angle of 2θ ± 0.2° has diffraction peaks at 5.75°, 7.09°, 8.42°, 9.07°, 9.74°, 11.53°, 12.80°, 13.28°, 14.08°, 15.03°, 15.34°, 16.01°, 16.67°, 16.89°, 17.31°, 18.21°, 18.46°, 19.13°, 19.53°, 20.02°, and 21.13°.

[0013] More preferably, the X-ray powder diffraction spectrum of Form II expressed at a diffraction angle of 2θ±0.2° is 5.75°, 7.09°, 8.42°, 9.07°, 9.74°, 10.83°, 11.53°, 12.80°, 13.28°, 14.08°, 14.65°, 15.03°, 15.34°, 16.01°, 16.67°, 16.89°, 17.31°, 18.21°, 18.46°, 19.13°, 19.53°, 20.02°, 21.13°, 21. There are diffraction peaks at .92°, 22.14°, 22.49°, 23.03°, 23.98°, 24.36°, 25.45°, 26.01°, 26.79°, 27.41°, 27.73°, 28.41°, 28.99°, 29.64°, 30.12°, 30.37°, 31.15°, 31.46°, 32.30°, 32.98°, 33.61°, 34.09°, 35.26°, 37.01°, 38.55°, 38.96°, and 39.68°.

[0014] Preferably, Form II has one or more of the following characteristics: The DSC curve of Form II has an endothermic peak at 50.1 ± 3 °C; The DSC curve of Form II has an endothermic peak at 127.8±3°C; The DSC curve of Form II has an endothermic peak at 244.6±3°C; The TGA curve of Form II shows a weight loss of 2.6±1% at 103.1±3°C; The TGA curve of Form II has a decomposition endothermic peak starting point at 359.7±3°C.

[0015] The X-ray powder diffraction spectrum of Form III expressed at a diffraction angle of 2θ±0.2° has diffraction peaks at 4.27°, 8.53°, 9.68°, 12.25°, 12.83°, 13.67°, 15.17°, 15.89°, 16.90°, and 19.10°.

[0016] Preferably, the X-ray powder diffraction spectrum of Form III expressed at a diffraction angle of 2θ±0.2° has diffraction peaks at 4.27°, 6.980°, 7.57°, 8.53°, 9.68°, 10.80°, 12.25°, 12.83°, 13.67°, 14.10°, 14.56°, 15.17°, 15.89°, 16.90°, 19.10°, 20.03°, 20.62°, 26.69°, and 30.63°.

[0017] More preferably, the X-ray powder diffraction spectrum of Form III expressed at a diffraction angle of 2θ±0.2° is 4.27°, 6.98°, 7.57°, 8.53°, 9.68°, 10.37°, 10.80°, 12.25°, 12.83°, 13.67°, 14.103°, 14.56°, 15.17°, 15.89°, 16.90°, 18.01 There are diffraction peaks at 19.10°, 20.03°, 20.62°, 21.93°, 22.51°, 23.17°, 24.36°, 24.68°, 25.43°, 26.69°, 28.74°, 29.42°, 30.63°, 31.99°, 33.39°, 34.09°, 35.95°, 37.07°, and 38.57°.

[0018] Preferably, Form III has one or more of the following characteristics: The DSC curve of Form III has an endothermic peak at 120.3 ± 3 °C; The TGA curve of Form III showed a weight loss of 7.5±1% at 128.6±3°C; The TGA curve of Form III has a decomposition endothermic peak starting point at 333.4±3°C.

[0019] The X-ray powder diffraction spectrum of Form IV expressed at a diffraction angle of 2θ±0.2° has diffraction peaks at 6.78°, 7.42°, 11.66°, 12.07°, 14.74°, 15.34°, 16.22°, 17.79°, 18.79°, and 21.09°.

[0020] Preferably, the X-ray powder diffraction spectrum of Form IV represented by a diffraction angle of 2θ±0.2° has diffraction peaks at 6.78°, 7.42°, 10.01°, 11.66°, 12.07°, 13.12°, 14.74°, 15.34°, 16.22°, 16.78°, 17.79°, 18.79°, 20.02°, 21.09°, 23.25°, and 24.28°.

[0021] More preferably, the X-ray powder diffraction spectrum of Form IV expressed at a diffraction angle of 2θ±0.2° has diffraction peaks at 6.78°, 7.42°, 10.01°, 11.66°, 12.07°, 13.12°, 14.74°, 15.34°, 16.22°, 16.78°, 17.79°, 18.79°, 20.02°, 21.09°, 22.53°, 23.25°, 24.28°, 25.19°, 25.94°, 26.89°, 27.92°, 28.85°, 30.10°, 30.92°, 33.02°, 34.67°, 35.26°, 36.45°, and 37.91°.

[0022] Preferably, Form IV has one or more of the following characteristics: The DSC curve of Form IV has an endothermic peak at 114.8 ± 3 °C; The DSC curve of Form IV has an endothermic peak at 150.7 ± 3 °C; The DSC curve of Form IV has an endothermic peak at 250.5 ± 3 °C; The TGA curve of Form IV showed a weight loss of 5.4±1% at 256.4±3°C; The TGA curve of Form IV has a decomposition endothermic peak starting point at 355.7±3°C.

[0023] The X-ray powder diffraction spectrum of Form V expressed at a diffraction angle of 2θ±0.2° has diffraction peaks at 7.77°, 10.83°, 12.17°, 12.72°, 13.07°, 14.00°, 15.21°, 15.71°, 16.36°, 17.45°, 19.06°, 21.15°, and 24.49°.

[0024] Preferably, the X-ray powder diffraction spectrum of Form V expressed at a diffraction angle of 2θ±0.2° has diffraction peaks at 5.57°, 6.26°, 6.91°, 7.77°, 9.48°, 9.96°, 10.83°, 11.24°, 12.17°, 12.72°, 13.07°, 14.00°, 14.59°, 15.21°, 15.71°, 16.36°, 17.45°, 19.06°, 20.45°, 20.73°, 21.15°, 24.49°, 27.05°, and 27.49°.

[0025] More preferably, the X-ray powder diffraction spectrum of Form V expressed at a diffraction angle of 2θ±0.2° is 5.57°, 6.26°, 6.91°, 7.77°, 9.48°, 9.96°, 10.83°, 11.24°, 12.17°, 12.72°, 13.07°, 14.00°, 14.58°, 15.21°, 15.71°, 16.36°, 17.45°, 19.06°, 20.45°, 20.73°, 2 There are diffraction peaks at 1.15°, 22.68°, 23.25°, 23.56°, 24.49°, 25.02°, 25.62°, 25.94°, 26.65°, 27.05°, 27.49°, 29.57°, 30.12°, 31.34°, 32.51°, 33.12°, 34.64°, 35.82°, 36.80°, and 37.56°.

[0026] Preferably, Form V has one or more of the following characteristics: The DSC curve of Form V has an endothermic peak at 115.3 ± 3 °C; The DSC curve of Form V has an endothermic peak at 155.3±3°C; The DSC curve of Form V has an endothermic peak at 169.6±3°C; The DSC curve of Form V has an endothermic peak at 254.9 ± 3 °C; The TGA curve of Form V shows a weight loss of 17.9±1% at 232.2±3℃; The TGA curve of Form V has a decomposition endothermic peak starting point at 330.3±3℃.

[0027] No obvious diffraction peaks were observed for amorphous form I, but rather a broad diffuse peak was observed.

[0028] Preferably, the amorphous form I has one or more of the following characteristics: The DSC curve of amorphous form I has an endothermic peak at 110.8 ± 3 °C; The DSC curve of amorphous form I shows the glass transition temperature at 265.3 ± 3 °C; The TGA curve of amorphous form I showed a weight loss of 6.2% at 189.3 ± 3 °C; The TGA curve of amorphous form I has a decomposition endothermic peak starting point at 326.5±3℃.

[0029] Amorphous II did not show obvious diffraction peaks, but rather showed a broad diffuse peak. Preferably, the amorphous form II has one or more of the following characteristics: The DSC curve of amorphous form II has an endothermic peak at 87.0 ± 3 °C; The DSC curve of amorphous II shows the glass transition temperature at 271.8 ± 3 °C; The TGA curve of amorphous II showed a weight loss of 3.6% at 120.0 ± 3 °C; The TGA curve of amorphous II has a decomposition endothermic peak starting point at 326.1±3℃.

[0030] The present invention provides a preparation method of various crystal forms of Sanguisorba officinalis saponin II sodium salt.

[0031] The preparation method of the crystal form I, crystal form III, and crystal form IV comprises the following steps: preparing a sodium salt solution of Sanguisorba officinalis saponin II, completely volatilizing the solvent in the solution, and precipitating a solid. The solvent is ethanol, methanol, or dimethyl sulfoxide.

[0032] Preferably, in the preparation methods of Form I, Form III, and Form IV, the mass concentration of the sodium salt solution of Sanguisorba officinalis saponin II is 2-200 mg / mL, and the solvent is volatilized by placing it in the open at 25-40°C.

[0033] The preparation method of the crystal form II comprises the following steps: taking the crystal form I and placing it in an environment of 22-28° C. and 75-85% humidity for 1-3 days.

[0034] The preparation method of the crystal form V comprises the following steps: preparing an ethylene glycol solution of Sanguisorba officinalis saponin II sodium salt, slowly adding isopropyl alcohol, stirring, filtering, and drying.

[0035] Preferably, in the preparation method of Form V, the mass concentration of the ethylene glycol solution of Sanguisorba officinalis saponin II sodium salt is 0.03-0.05 g / mL, the added volume of isopropanol is 4-8 times that of ethylene glycol, the stirring time is 1-3 h, and the drying temperature is 35-45°C.

[0036] The present invention provides a method for preparing an amorphous substance of Sanguisorba officinalis saponin II sodium salt.

[0037] The preparation method of the amorphous form I comprises the following steps: preparing a methanol solution of sodium salt of Sanguisorba officinalis saponin II and spray drying the solution.

[0038] Preferably, in the preparation method of amorphous form I, the mass concentration of the methanol solution of sanguisorbaside II sodium salt is 0.02-0.03 g / mL, the inlet temperature of the spray drying is 150-170°C, and the outlet temperature is 70-80°C.

[0039] The preparation method of the amorphous form II comprises the following steps: preparing an ethylene glycol solution of sodium salt of Sanguisorba officinalis saponin II, slowly adding isopropyl alcohol, stirring, filtering, and drying.

[0040] Preferably, in the preparation method of amorphous form II, the mass concentration of the ethylene glycol solution of sanguisorbaside II sodium salt is 0.03-0.05 g / mL, the added volume of isopropanol is 4-8 times that of ethylene glycol, the stirring time is 1-3h, and the drying temperature is 140-160°C.

[0041] The present invention also provides the use of the crystal or amorphous substance of Sanguisorba officinalis saponin II sodium salt in the preparation of medicines.

[0042] Preferably, the drug is a drug for treating lung diseases.

[0043] The present invention also provides a pharmaceutical preparation comprising the crystal or amorphous form of the sodium salt of Sanguisorba officinalis saponin II and one or more pharmaceutically acceptable carriers or excipients.

[0044] Preferably, the pharmaceutical preparation is an inhalation preparation.

[0045] The present invention also provides a method for controlling the particle size of ZYG-II-Na crystals or amorphous materials used as inhalation preparations, which specifically includes grinding, air flow crushing, spray drying, and anti-solvent method.

[0046] Beneficial effects of the present invention: (1) The polymorphic and amorphous forms of ZYG-II-Na of the present invention improve the solubility of the compound in water; (2) The solid form significantly improves the bioavailability of ZYG-Ⅱ-Na, thereby enhancing its efficacy; (3) By improving the physicochemical properties and in vivo absorption performance, the polymorphic and amorphous forms of ZYG-Ⅱ-Na of the present invention provide strong support for its application in the pulmonary administration route. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 is the PXRD spectrum of Form I in Example 1; Figure 2 is the DSC spectrum of Form I in Example 1; Figure 3 This is the TGA spectrum of Form I in Example 1; Figure 4is the PXRD spectrum of Form II in Example 2; Figure 5 This is the DSC spectrum of Form II in Example 2; Figure 6 This is the TGA spectrum of Form II in Example 2; Figure 7 is the PXRD spectrum of Form III in Example 3; Figure 8 This is the DSC spectrum of Form III in Example 3; Figure 9 This is the TGA spectrum of Form III in Example 3; Figure 10 is the PXRD spectrum of Form IV in Example 4; Figure 11 is the DSC spectrum of Form IV in Example 4; Figure 12 is the TGA spectrum of Form IV in Example 4; Figure 13 is the PXRD spectrum of Form V in Example 5; Figure 14 is the DSC spectrum of Form V in Example 5; Figure 15 is the TGA spectrum of Form V in Example 5; Figure 16 is the PXRD spectrum of amorphous form I in Example 6; Figure 17 is the DSC spectrum of amorphous form I in Example 6; Figure 18 is the TGA spectrum of amorphous form I in Example 6; Figure 19 is the PXRD spectrum of amorphous form II in Example 7; Figure 20 is the DSC spectrum of amorphous form II in Example 7; Figure 21 is the TGA spectrum of amorphous form II in Example 7; Figure 22 is the NGI deposition distribution diagram of each solid form in Example 8; Figure 23 This is the DVS spectrum of Form I in Example 9; Figure 24 This is the DVS spectrum of Form II in Example 9; Figure 25 This is the DVS spectrum of Form III in Example 9; Figure 26 This is the DVS spectrum of amorphous form I in Example 9; Figure 27is the DVS spectrum of amorphous form II in Example 9; Figure 28 is the PXRD spectrum of Form I in Example 10 at 40° C. and 40% RH; Figure 29 This is the PXRD spectrum of Form II in Example 10 at 40°C and 40% RH; Figure 30 This is the PXRD spectrum of Form III in Example 10 at 40°C and 40% RH; Figure 31 This is the PXRD spectrum of amorphous form I in Example 10 at 40°C and 40% RH; Figure 32 This is the PXRD spectrum of amorphous form II in Example 10 at 40°C and 40% RH; Figure 33 is the PXRD spectrum of Form I in Example 10 at 40° C. and 75% RH; Figure 34 This is the PXRD spectrum of Form II in Example 10 at 40°C and 75% RH; Figure 35 This is the PXRD spectrum of Form III in Example 10 at 40°C and 75% RH; Figure 36 This is the PXRD spectrum of amorphous form I in Example 10 at 40°C and 75% RH; Figure 37 This is the PXRD spectrum of amorphous form II in Example 10 at 40°C and 75% RH; Figure 38 1 is the dissolution curve of each solid form of ZYG-II-Na in water in Example 11; Figure 39 1 is the dissolution curve of each solid form of ZYG-II-Na in artificial lung fluid in Example 11; Figure 40 This is the drug-time curve of ZYG-II-Na administered via the tail vein in Example 12; Figure 41 This is the drug-time curve of ZYG-Ⅱ-Na in Example 12 when administered by oral gavage and dry powder inhalation. DETAILED DESCRIPTION

[0048] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0049] Unless otherwise specified, all reagents involved in the embodiments of the present invention are commercially available products and can be purchased through commercial channels.

[0050] 1. Test Method 1. X-ray powder diffraction (PXRD) test The detection parameters were as follows: monochromatic Cu Kα radiation (λ = 1.54 Å), tube voltage: 40 kV, current: 40 mA, scanning range: 3–40°, step size: 0.02°, and scanning speed: 0.1 s / step.

[0051] 2. Thermogravimetric analysis (TGA) Accurately weigh 5-10 mg of powdered sample and place it in a platinum crucible in a thermogravimetric analyzer. Under a nitrogen atmosphere (flow rate approximately 50 ml / min), heat the sample from 30°C to 550°C at a rate of 10 K / min. Record the change in sample mass percentage versus temperature.

[0052] 3. Differential Scanning Calorimetry (DSC) Accurately weigh 5-10 mg of powder sample and place it in a sealed aluminum crucible. Under a nitrogen atmosphere, heat the sample from 40°C to 300°C at a rate of 10 K / min, and record the heat flow change.

[0053] 2. Preparation of various solid forms of ZYG-Ⅱ-Na The structural formula of compound I (ZYG-Ⅱ-Na) is as follows: .

[0054] Example 1: Preparation of Form I Take 20 mg of compound I, add it to 7 mL of ethanol, heat it to 40 ° C and stir to dissolve it. Open it under the condition of keeping warm to allow the ethanol to evaporate completely, precipitate the solid, take a sample for testing, and mark it as crystal form I. The test results are shown in Figure 1 、 Figure 2 and Figure 3 . Figure 2 The DSC results show that the sample has an endothermic peak near 130.6℃ and an endothermic peak near 244.5℃, which are related to the change of intermolecular force or stress release at high temperature. Figure 3 The TGA results show that the sample begins to decompose at around 345.4°C. The PXRD diffraction peak data of Form I are shown in Table 1.

[0055] Table 1 PXRD diffraction peak data of Form I

[0056] Example 2: Preparation of Form II Take 20 mg of Form I, place it in an environment of 25°C and 80% humidity for 2 days, take a sample for testing, and mark it as Form II. Figure 4 、 Figure 5 and Figure 6 . Figure 5 The DSC results show that the sample has a desolvation crystallization endothermic peak near 50.1°C, an endothermic peak near 127.8°C, and an endothermic peak near 244.6°C. The endothermic peaks at 127.8°C and 244.6°C are related to changes in intermolecular forces or stress release at high temperatures. Figure 6 The TGA results show that the sample loses 2.6% weight when heated from 40°C to 103.1°C, and decomposition begins around 359.7°C. The PXRD diffraction peak data for Form II are shown in Table 2.

[0057] Table 2 PXRD diffraction peak data of Form II

[0058] Example 3: Preparation of Form III Take 200 mg of compound I, add it to 10 mL of methanol, dissolve it by ultrasonication at 25°C, open it at 25°C to allow the methanol to completely evaporate, and precipitate the solid. Samples were taken for testing and marked as Form III. The test results are shown in Figure 7 、 Figure 8 and Figure 9 . Figure 8 The DSC results showed that the sample had an endothermic peak near 120.3 °C, which was related to the change of intermolecular forces or stress release at high temperature. Figure 9 The TGA results show that the sample loses 7.5% weight when heated from 40°C to 128.6°C, and decomposition begins around 333.4°C. The PXRD diffraction peak data for Form III are shown in Table 3.

[0059] Table 3 PXRD diffraction peak data of Form III

[0060] Example 4: Preparation of Form IV Take 200 mg of compound I, add it to 1 mL of dimethyl sulfoxide, dissolve it by ultrasonication at 25°C, and open it at 25°C to allow dimethyl sulfoxide to completely evaporate. The solid precipitated was sampled and tested, and marked as Form IV. The test results are shown in Figure 10 、 Figure 11 and Figure 12 . Figure 11 The DSC results show that the sample has a desolvation crystallization endothermic peak near 114.8℃, an endothermic peak near 150.3℃, and an endothermic peak near 250.5℃. The endothermic peaks at 150.7℃ and 250.5℃ are related to the change of intermolecular force or stress release at high temperature. Figure 12The TGA results show that the sample loses 5.4% weight when heated from 40°C to 256.4°C, and decomposition begins around 355.7°C. The PXRD diffraction peak data for Form IV are shown in Table 4.

[0061] Table 4 PXRD diffraction peak data of Form IV

[0062] Example 5: Preparation of Form V Take 1g of compound I, add it to 25mL of ethylene glycol, dissolve it by ultrasonication at 25℃, slowly add 150mL of isopropanol dropwise to the resulting solution, stir it for 2h, filter it with suction, dry the filter cake in an oven at 40℃, take a sample for testing, and mark it as Form V. The test results are shown in Figure 13 and Figure 14 and Figure 15 . Figure 14 The DSC results showed that the sample had endothermic peaks near 115.3℃, 155.3℃, 169.6℃ and 254.9℃. Figure 15 The TGA results show that the sample loses 17.9% weight when heated from 40°C to 232.2°C, and decomposition begins around 330.3°C. The PXRD diffraction peak data for Form V are shown in Table 5.

[0063] Table 5 PXRD diffraction peak data of Form V

[0064] Example 6: Preparation of Amorphous Form I 4.5 g of compound I was added to 180 mL of methanol and dissolved by ultrasonication at 25 °C. The mixture was dried using a spray dryer at an inlet temperature of 160.0 °C and an outlet temperature of 73.8 °C. A sample was taken for testing and marked as amorphous I. The test results are shown in FIG. Figure 16 、 Figure 17 and Figure 18 . Figure 12 The DSC results showed that the sample had an endothermic peak near 110.8°C and a glass transition temperature (Tg) of 265.3°C. Figure 12 The TGA results showed that the sample lost 6.2% of its weight when heated from 40°C to 189.3°C, and the sample began to decompose around 326.5°C.

[0065] Example 7: Preparation of Amorphous Form II Take 1g of compound I, add it to 25mL of ethylene glycol, dissolve it by ultrasonic at 25℃, slowly add 150mL of isopropanol dropwise to the resulting solution, stir for 2h, filter it, dry the filter cake in an oven at 150℃, take a sample for testing, and mark it as amorphous II. Figure 19 and Figure 20 and Figure 21 . Figure 20 The DSC results showed that the sample had an endothermic peak near 87.0℃ and a glass transition temperature (Tg) of 271.8℃. Figure 21 The TGA results showed that the sample lost 3.6% weight when heated from 40°C to 120.0°C, and the sample began to decompose around 326.1°C.

[0066] Example 8: Particle Size Control for Inhalation Drug Delivery Each crystalline form and amorphous form were sieved through 40 mesh and subjected to air flow milling at an air inlet pressure of 6.5 bar, a milling pressure of 6.0 bar, and a feed rate of 2 rpm. Particle size distribution was determined by laser diffraction.

[0067] The aerodynamic size and in vitro deposition efficiency of ZYG-Ⅱ-Na particles in various solid forms were determined using the Next Generation Impactor (NGI). The NGI, comprised of an artificial throat, a pre-separator, a seven-stage cascade impactor, and a micro-orifice collector (MOC), allows for fractional collection of particles based on their aerodynamic properties under simulated inhalation flow conditions, enabling a comprehensive evaluation of their deposition behavior. No. 3 hydropropyl methylcellulose (HPMC) capsules were used as carriers. Each capsule was loaded with 10 mg of ZYG-Ⅱ-Na powder and subsequently placed individually into the inhalation device. After puncture, the capsules were tightly connected to the NGI system and inhalation testing was performed under standard conditions of an airflow rate of 60 L / min and a run time of 4.0 s. To minimize secondary rebound or migration of particles on the impactor plates and ensure accurate particle deposition at the corresponding impactor levels, the surfaces of the NGI impactor plates (stages 1 to 7) and the MOC were pre-coated with a 1% silicone oil-n-hexane solution and allowed to evaporate naturally. Three capsules were tested in duplicate for each solid ZYG-Ⅱ-Na sample. After the experiment, the NGI impactor plates and capsules were thoroughly rinsed with ethanol. All deposited powder was recovered and, after appropriate treatment, analyzed by HPLC. The particle deposition distribution at each impactor stage was calculated, along with the capsule emptying and recovery rates. Each experiment was repeated three times to ensure data reliability and reproducibility.

[0068] The experimental data were processed using Copley Inhalation Dosage Test Data Analysis Software (CITDAS, Version 3.10) to calculate the particle deposition ratios at each level and to derive key aerodynamic parameters, including the fine particle fraction (FPF), mass median aerodynamic diameter (MMAD), and geometric standard deviation (GSD), for a comprehensive evaluation of the pulmonary delivery performance of different solid forms of ZYG-Ⅱ-Na.

[0069] Particle size results: shown in Table 6.

[0070] Table 6 Crystalline and amorphous particle sizes of ZYG-II-Na for inhalation

[0071] Aerodynamic evaluation results: Figure 22 , Table 7. All solid forms exhibited excellent aerodynamic performance. The MMAD of the powder inhaler ranged from 1.87 to 3.21 μm, meeting the particle size requirements for pulmonary delivery. GSD is used to describe the uniformity of the particle size distribution. A smaller GSD indicates a narrower particle size distribution, which helps improve pulmonary deposition efficiency. In this study, the GSD values ​​of different solid form DPI preparations were all less than 2, indicating that their particle size distribution was narrow. FPF, as one of the important evaluation indicators of powder inhalers, represents the mass percentage of particles with a size less than 5 μm in the total released amount. The study found that all solid forms exhibited high FPF values, among which the FPF of crystalline forms I, II, and III amorphous form I exceeded 70%, and the FPF of amorphous form II was 32.75 ± 1.62%, both meeting the requirements for pulmonary delivery. In addition, the capsule emptying rate and recovery rate of all solid forms were greater than 90%.

[0072] Table 7 Aerodynamic parameters of the crystalline and amorphous forms of ZYG-Ⅱ-Na for inhalation (n = 3)

[0073] Example 9: Dynamic Water Vapor Adsorption Test Dynamic Vapor Sorption (DVS) testing was performed on ZYG-Ⅱ-Na Form I, Form II, Form III, Amorphous Form I, and Amorphous Form II. Approximately 20 mg of powder sample was placed in the sample tray of the dynamic vapor sorption instrument for testing. The experiment was conducted at 25°C, with the RH increasing from 0% to 95% in 5% increments, and then decreasing from 95% to 0% in the same gradient. The results are shown in the figure below. Figure 23-27As shown, the hygroscopicity of ZYG-Ⅱ-Na from weak to strong is crystal form III, crystal form II, crystal form I, amorphous form II, and amorphous form I, and the hygroscopicity of the crystal form is weaker than that of the amorphous form.

[0074] Example 10: Stability Test The stability of ZYG-Ⅱ-Na crystal form I, crystal form II, crystal form III, amorphous form I and amorphous form II was tested, and each solid form of ZYG-Ⅱ-Na was placed in an environment with a relative humidity (RH) of 75% and 40% and a constant temperature of 40°C for accelerated stability testing. The 75% humidity condition was obtained by controlling the humidity in a sealed container with saturated sodium chloride (NaCl), and the humidity was monitored in real time by a hygrometer to ensure environmental stability. The 40% humidity condition was controlled by a drug stability test chamber. The samples were stored under the above conditions on the 10th, 20th, 30th, 60th and 90th days of storage, and samples were taken and their stability over time was evaluated by PXRD analysis. The results are shown in the figure. Figure 28-37 As shown, at 40°C, 40% RH, the PXRD patterns of Form I, Form III, Amorphous Form I, and Amorphous Form II remained unchanged for more than 90 days, and Form II underwent a crystal form transformation after 10 days. At 40°C, 75% RH, the PXRD patterns of Form III remained unchanged for more than 90 days, and Form I, Form II, Amorphous Form I, and Amorphous Form II underwent a crystal form transformation after 10 days.

[0075] Example 11: Solubility Test The solubility of ZYG-Ⅱ-Na crystal form I, crystal form II, crystal form III, amorphous form I and amorphous form II was tested in pure water and pH 7.4 Gamble's solution (artificial lung fluid) at a speed of 100 rpm and a temperature of 37±0.5°C. Figure 38 、 39 As shown, the maximum solubilities of ZYG-Ⅱ-Na in water are 4.5 mg / mL, 4.6 mg / mL, 17.1 mg / mL, 24.2 mg / mL and 15.9 mg / mL, respectively; and the maximum solubilities in artificial lung fluid are 0.20 mg / mL, 0.20 mg / mL, 0.62 mg / mL, 0.56 mg / mL and 0.53 mg / mL, respectively. The maximum solubilities of form III, amorphous form I and amorphous form II exceed those of form I and form II. Amorphous form I is better than other forms in maintaining supersaturation in water, and amorphous form II is better than other forms in maintaining supersaturation in artificial lung fluid.

[0076] Example 12: Pharmacokinetic Study The pharmacokinetics of ZYG-Ⅱ-Na crystal forms I, II, III, amorphous form I, and II were investigated in 42 male Sprague-Dawley rats, randomly divided into seven groups of six rats each. All animals had free access to water during the experiment and were fasted for 12 hours before dosing. One group of rats received a 10 mg / kg dose of ZYG-Ⅱ-Na (form I) solution via intravenous injection (iv); another group received the same dose of ZYG-Ⅱ-Na (form I) via intragastric gavage (ig). The remaining five groups received 10 mg / kg doses of various solid forms of ZYG-Ⅱ-Na, including crystal forms I, II, III, amorphous form I, and II, via dry powder inhaler. After administration, 0.2 ml of whole blood was collected from the retroorbital venous plexus at 0.083, 0.25, 0.5, 0.75, 1, 2, 4, 8, 10, and 24 hours for pharmacokinetic analysis. The whole blood was centrifuged at 3000 rpm for 10 minutes (4°C) to separate the plasma and test the concentration. Figure 40 、 41 As shown in Tables 8 and 9, the biological exposure level of DPI administration is higher than that of oral gavage. The bioavailability of DPI administration in different solid forms is amorphous form II, amorphous form I, crystalline form III, crystalline form II, and crystalline form I. These characteristics provide potential prospects for their application in drug development.

[0077] Table 8 Pharmacokinetic parameters of the crystalline forms and amorphous form of ZYG-Ⅱ-Na in SD rats via iv, ig and DPI

[0078] AUC is the area under the 24-hour drug-dose curve, C max is the maximum concentration, F is the relative bioavailability, and Fabs is the absolute bioavailability.

[0079] Table 9 Pharmacokinetic parameters of the crystalline forms and amorphous form of ZYG-Ⅱ-Na in SD rats via iv, ig and DPI

[0080] MRT (0-t) is the average residence time, t 1 / 2 is the half-life, T max Peak time.

[0081] Finally, it should be noted that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art will be able to modify the technical solutions described in the aforementioned embodiments or substitute equivalents for some of the technical features. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.

Claims

1. A crystalline or amorphous substance of Sanguisorba officinalis saponin II sodium salt, characterized in that: The chemical structural formula of the sodium salt of Sanguisorba officinalis saponin II is: ， The crystal form of the sodium salt of Sanguisorba officinalis saponin II is crystal form I, crystal form II, crystal form III, crystal form IV or crystal form V, and the amorphous form of the sodium salt of Sanguisorba officinalis saponin II is amorphous form I or amorphous form II; The diffraction peaks of the crystalline form I in the X-ray powder diffraction spectrum, expressed as a diffraction angle of 2θ±0.2°, include: 6.12°, 6.89°, 10.61°, 12.17°, 12.85°, 13.30°, 14.39°, 14.57°, 15.48°, 15.89°, 16.98°, 18.26°, and 22.63°; The diffraction peaks of the crystalline form II in the X-ray powder diffraction spectrum, expressed as a diffraction angle of 2θ±0.2°, include: 5.75°, 7.09°, 9.74°, 11.53°, 13.28°, 14.08°, 15.03°, 15.40°, 18.21°, 18.46°, and 21.13°; The diffraction peaks of the crystalline form III in the X-ray powder diffraction spectrum, expressed as a diffraction angle of 2θ±0.2°, include: 4.27°, 8.53°, 9.68°, 12.25°, 12.83°, 13.67°, 15.17°, 15.89°, 16.90°, and 19.10°; The diffraction peaks of the crystalline form IV in the X-ray powder diffraction spectrum, expressed as a diffraction angle of 2θ±0.2°, include: 6.78°, 7.42°, 11.66°, 12.07°, 14.74°, 15.34°, 16.22°, 17.79°, 18.79°, and 21.09°; The diffraction peaks of the crystalline form V in the X-ray powder diffraction spectrum, expressed as a diffraction angle of 2θ±0.2°, include: 7.77°, 10.83°, 12.17°, 12.72°, 13.07°, 14.00°, 15.21°, 15.71°, 16.36°, 17.45°, 19.06°, 21.15°, and 24.49°; The amorphous form I or amorphous form II has no obvious diffraction peak in the X-ray powder diffraction spectrum.

2. The crystal or amorphous substance of Sanguisorbaside II sodium salt according to claim 1, characterized in that: The crystalline form I has one or more of the following characteristics: A1, diffraction peaks of Form I in the X-ray powder diffraction spectrum, expressed as 2θ±0.2° diffraction angles include: 6.12°, 6.89°, 10.61°, 12.17°, 12.85°, 13.30°, 14.39°, 14.57°, 15.48°, 15.89°, 16.55°, 16.98°, 18.26°, 19.18°, and 22.63°; The DSC curve of A2 and Form I has an endothermic peak at 130.6±3°C; The DSC curve of A3 and Form I has an endothermic peak at 244.5 ± 3 °C; The crystal form II has one or more of the following characteristics: B1, the diffraction peaks of Form II in the X-ray powder diffraction spectrum, expressed as 2θ±0.2° diffraction angles include: 5.75°, 7.09°, 8.42°, 9.07°, 9.74°, 11.53°, 12.80°, 13.28°, 14.08°, 15.03°, 15.34°, 16.01°, 16.67°, 16.89°, 17.31°, 18.21°, 18.46°, 19.13°, 19.53°, 20.02°, and 21.13°; The DSC curve of B2 and Form II has an endothermic peak at 50.1 ± 3 °C; The DSC curve of B3 and Form II has an endothermic peak at 127.8 ± 3 °C; The DSC curve of B4 and Form II has an endothermic peak at 244.6 ± 3 °C; The crystalline form III has one or more of the following characteristics: C1, the diffraction peaks of Form III in the X-ray powder diffraction spectrum, expressed as 2θ±0.2° diffraction angles include: 4.27°, 6.980°, 7.57°, 8.53°, 9.68°, 10.80°, 12.25°, 12.83°, 13.67°, 14.10°, 14.56°, 15.17°, 15.89°, 16.90°, 19.10°, 20.03°, 20.62°, 26.69°, and 30.63°; The DSC curve of C2, Form III, has an endothermic peak at 120.3 ± 3 °C; The crystalline form IV has one or more of the following characteristics: Diffraction peaks of Form IV in the X-ray powder diffraction spectrum, expressed as 2θ±0.2° diffraction angles, include: 6.78°, 7.42°, 10.01°, 11.66°, 12.07°, 13.12°, 14.74°, 15.34°, 16.22°, 16.78°, 17.79°, 18.79°, 20.02°, 21.09°, 23.25°, and 24.28°; The DSC curve of D2 and Form IV has an endothermic peak at 114.8 ± 3 °C; The DSC curve of D3 and Form IV has an endothermic peak at 150.7 ± 3 °C; The DSC curve of D4 and Form IV has an endothermic peak at 250.5 ± 3 °C; The crystalline form V has one or more of the following characteristics: E1, the diffraction peaks of Form V in the X-ray powder diffraction spectrum, expressed as 2θ±0.2° diffraction angles include: 5.57°, 6.26°, 6.91°, 7.77°, 9.48°, 9.96°, 10.83°, 11.24°, 12.17°, 12.72°, 13.07°, 14.00°, 14.59°, 15.21°, 15.71°, 16.36°, 17.45°, 19.06°, 20.45°, 20.73°, 21.15°, 24.49°, 27.05°, and 27.49°; The DSC curves of E2 and Form V have an endothermic peak at 115.3 ± 3 °C; The DSC curves of E3 and Form V have an endothermic peak at 155.3 ± 3 °C; The DSC curve of E4 and Form V has an endothermic peak at 169.6 ± 3 °C; The DSC curve of E5 and Form V has an endothermic peak at 254.9 ± 3 °C; The amorphous form I has one or more of the following characteristics: The DSC curves of F1 and amorphous form I have an endothermic peak at 110.8 ± 3 °C; The DSC curve of F2 and amorphous I shows the glass transition temperature at 265.3 ± 3 °C; The amorphous II has one or more of the following characteristics: The DSC curves of G1 and amorphous II have an endothermic peak at 87.0 ± 3 °C; The DSC curve of G2 and amorphous II shows the glass transition temperature at 271.8±3℃.

3. The crystal or amorphous substance of Sanguisorbaside II sodium salt according to claim 1, characterized in that: The crystalline form I also has one or more of the following characteristics: a1. The PXRD spectrum of Form I is shown in Figure 1; a2. The DSC spectrum of Form I is shown in Figure 2; a3. The TGA spectrum of Form I is shown in Figure 3; The crystalline form II also has one or more of the following characteristics: b1, the PXRD spectrum of Form II is shown in Figure 4; b2. The DSC spectrum of Form II is shown in Figure 5; b3. The TGA spectrum of Form II is shown in Figure 6; The crystalline form III also has one or more of the following characteristics: c1, the PXRD spectrum of Form III is shown in Figure 7; c2. The DSC spectrum of Form III is shown in Figure 8; c3. The TGA spectrum of Form III is shown in Figure 9; The crystalline form IV also has one or more of the following characteristics: d1, the PXRD spectrum of Form IV is shown in Figure 10; d2, the DSC spectrum of Form IV is shown in Figure 11; d3, the TGA spectrum of Form IV is represented in Figure 12; The crystalline form V also has one or more of the following characteristics: e1, the PXRD spectrum of Form V is shown in Figure 13; e2. The DSC spectrum of Form V is shown in Figure 14; e3. The TGA spectrum of Form V is shown in Figure 15; The amorphous form I also has one or more of the following characteristics: f1, the PXRD spectrum of amorphous I is represented in Figure 16; f 2. The DSC spectrum of amorphous form I is represented in Figure 17; f 3. The TGA spectrum of amorphous form I is represented in Figure 18; The amorphous II also has one or more of the following characteristics: The PXRD patterns of g1 and amorphous II are shown in Figure 19 ; g2, the DSC spectrum of amorphous form II is represented in Figure 20; The TGA spectrum of g3, amorphous II is shown in Figure 21.

4. A method for preparing the crystals of Sanguisorbaside II sodium salt according to claim 1, characterized in that: The preparation method of the crystal form I, crystal form III, and crystal form IV comprises the following steps: preparing a solution of sodium salt of Sanguisorba officinalis saponin II, volatilizing the solvent in the solution to precipitate a solid; the solvent is ethanol, methanol, or dimethyl sulfoxide; The preparation method of the crystal form II comprises the following steps: taking the crystal form I and placing it in an environment of 22-28° C. and 75-85% humidity for 1-3 days; The preparation method of the crystal form V comprises the following steps: preparing an ethylene glycol solution of Sanguisorba officinalis saponin II sodium salt, slowly adding isopropyl alcohol, stirring, filtering, and drying.

5. The method for preparing the crystal according to claim 4, wherein: In the preparation methods of Form I, Form III, and Form IV, the mass concentration of the Sanguisorbaside II sodium salt solution is 2-200 mg / mL, and the solvent is evaporated by placing it in the open at 25-40°C; In the preparation method of crystal form V, the mass concentration of the ethylene glycol solution of Sanguisorba officinalis saponin II sodium salt is 0.03-0.05 g / mL, the added volume of isopropanol is 4-8 times that of ethylene glycol, the stirring time is 1-3 hours, and the drying temperature is 35-45°C.

6. A method for preparing the amorphous substance of Sanguisorba officinalis saponin II sodium salt according to claim 1, characterized in that: The preparation method of the amorphous form I comprises the following steps: preparing a methanol solution of sodium salt of Sanguisorba officinalis saponin II and spray drying; The preparation method of the amorphous form II comprises the following steps: preparing an ethylene glycol solution of sodium salt of Sanguisorba officinalis saponin II, slowly adding isopropyl alcohol, stirring, filtering, and drying.

7. The method for preparing the amorphous substance according to claim 6, wherein: In the preparation method of amorphous form I, the mass concentration of the methanol solution of sanguisorbaside II sodium salt is 0.02-0.03 g / mL, the inlet temperature of the spray drying is 150-170°C, and the outlet temperature is 70-80°C; In the preparation method of amorphous form II, the mass concentration of the ethylene glycol solution of sanguisorbaside II sodium salt is 0.03-0.05 g / mL, the added volume of isopropanol is 4-8 times that of ethylene glycol, the stirring time is 1-3 hours, and the drying temperature is 140-160°C.

8. Use of the crystal or amorphous substance of the sodium salt of Sanguisorba officinalis saponin II according to claim 1 in the preparation of medicines.

9. A pharmaceutical preparation, characterized in that: The invention comprises the crystal or amorphous substance of the sodium salt of Sanguisorba officinalis saponin II according to claim 1, and one or more pharmaceutically acceptable carriers or excipients.

10. A method for controlling the particle size of a crystalline or amorphous form of Sanguisorba officinalis saponin II sodium salt as an inhalation preparation, characterized in that: The method includes any one or more of grinding, air flow milling, spray drying and anti-solvent method.

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