A nano-crystal suspension of galangin and a preparation method thereof

Abstract

The application discloses a nano-crystal suspension of rhizoxin, which comprises rhizoxin, a stabilizer and water as effective components, and the average particle size of the rhizoxin is 100-150 nm. The application also provides a preparation process. The preparation process of the application is stable and controllable, has excellent reproducibility, and is suitable for industrial amplification production by adopting a reverse-solvent precipitation-high-pressure homogenization method. The dissolution rate of the nano-crystal suspension of rhizoxin is significantly improved compared with that of the raw material, the bioavailability is improved, and an ideal solution is provided for the delivery of the poorly soluble drug rhizoxin.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to a galangin nanocrystal suspension and its preparation method. Background Technology

[0002] Galangin is a flavonoid active ingredient extracted from the roots and stems of the ginger family herbaceous plant *Alpinia galanga*. It possesses various pharmacological effects, including antitumor, anti-inflammatory, antioxidant, and antibacterial properties, and is of significant value in new drug development. However, galangin has poor water solubility and low bioavailability, severely limiting its clinical application. Currently, technologies such as microemulsions, nanoemulsions, and micelles are used to improve the solubility of galangin, but problems such as low drug loading, complex preparation processes, and poor stability still exist.

[0003] Patent CN114259473A discloses a galangin nanolipid assembly that achieves high encapsulation efficiency while effectively reducing particle size, improving the skin affinity of galangin nanosolution, and increasing skin permeability. However, the nanolipid assembly has a low drug loading capacity and a complex preparation process, which is not conducive to industrial production.

[0004] The literature "Formulation optimization and characterization of galangin-PLGA nanoparticles (Journal of Hunan Medical College, 2021, 27(11))" discloses a galangin-glycolic acid copolymer nanoparticle with a particle size of 249 nm and a high encapsulation efficiency. However, the drug loading of this nanoparticle is low, and galangin is highly hydrophobic, resulting in unstable encapsulation efficiency. In addition, the excipient PLGA may have certain potential toxicity, such as degradation into lactic acid in vivo, which poses a certain risk of inflammation.

[0005] Drug nanocrystals are nanoscale colloidal dispersion systems composed of pure drug particles and a small amount of stabilizer. They can significantly improve the solubility and bioavailability of poorly soluble drugs, and offer high drug loading, controllable particle size, and simple preparation methods, making them one of the most effective technologies for addressing the drug-like properties of poorly soluble drugs. Common preparation methods for nanocrystals include antisolvent precipitation, high-pressure homogenization, wet milling, or a combination of multiple technologies. Nanocrystals obtained by different methods vary significantly in particle size, uniformity, stability, and industrial feasibility.

[0006] Currently, no patent technology solutions for the preparation of nanocrystals using galangin as a model drug have been disclosed. Summary of the Invention

[0007] Purpose of the invention: The purpose of this invention is to solve the problems of poor water solubility and low bioavailability of galangin, and to provide a pharmaceutical composition with controllable particle size, uniform distribution and high dissolution rate, as well as its preparation method.

[0008] To achieve the above objectives, the present invention provides a galangin nanocrystal suspension comprising galangin as an active ingredient, a stabilizer, and water, wherein the average particle size of the galangin is 100-150 nm.

[0009] Furthermore, the stabilizer is selected from one or more of poloxamer 407, polyvinylpyrrolidone K30, hydroxypropyl methylcellulose, poloxamer 188, and vitamin E polyethylene glycol succinate.

[0010] Furthermore, the mass ratio of galangin to stabilizer is 1:0.5 to 1:2.

[0011] Furthermore, the mass ratio of galangin to stabilizer is 1:1.

[0012] Furthermore, the stabilizer is a compound stabilizer composed of poloxamer 407 and polyvinylpyrrolidone K30 in a 1:1 mass ratio. The stabilizer maintains stability primarily through electrostatic repulsion or steric hindrance. Poloxamer 407 itself possesses both steric hindrance and partial electrostatic repulsion, but when used alone, its adsorption rate is slow and its surface coverage is not dense. When compounded with polyvinylpyrrolidone K30, it can achieve synergistic stabilization by leveraging the rapid adsorption characteristics of polyvinylpyrrolidone K30.

[0013] Furthermore, the method for preparing the suspension includes the step of preparing a galangin nanocrystal suspension by using antisolvent precipitation-ultrasonic method or antisolvent precipitation-high pressure homogenization method with galangin, stabilizer and water.

[0014] Furthermore, the preparation method of the suspension is an antisolvent precipitation-high pressure homogenization method, specifically including the following steps:

[0015] (1) Alpinin was dissolved in an organic solvent to prepare the organic phase of the drug;

[0016] (2) Dissolve the stabilizer in purified water to obtain an aqueous phase;

[0017] (3) The organic phase was injected into the aqueous phase under high-speed shearing conditions to obtain a crude suspension;

[0018] (4) The coarse suspension was placed in a high-pressure homogenizer for circulation homogenization;

[0019] (5) Remove the organic solvent and filter to obtain galangin nanocrystal suspension.

[0020] Furthermore, the organic solvent is anhydrous ethanol, the concentration of galangin in the organic phase is 10 mg / mL, and the volume ratio of the aqueous phase to the organic phase is 8:1.

[0021] Furthermore, in step 3), the high-speed shearing speed is 800 rpm, and in step 4), the high-pressure homogenization pressure is 1000~1500 bar, and the number of cycles is 5~15.

[0022] Beneficial effects:

[0023] The galangin nanocrystal suspension prepared by this invention has a PDI (Polydispersity Index, used to describe the uniformity of particle size in a suspension) of less than 0.3, indicating that the particle size is controllable and the distribution is uniform. Compared with the prior art, it has the following advantages:

[0024] (1) Through the synergistic effect between the compounded stabilizers, the nanocrystals have a uniform particle size distribution, high stability, and high drug loading, while the amount of excipients used is relatively small and the safety is high.

[0025] (2) The preparation process of the present invention is stable and controllable, with excellent reproducibility. The anti-solvent precipitation-high pressure homogenization method is suitable for industrial scale-up production.

[0026] (3) The dissolution rate of the galangin nanocrystal suspension of the present invention is significantly improved compared with that of the raw drug, and the bioavailability is improved, providing an ideal solution for the delivery of the poorly soluble drug galangin. Attached Figure Description

[0027] Figure 1 This is a transmission electron microscope image of galangin nanocrystals from Example 1.

[0028] Figure 2 This is the external release curve of galangin nanocrystals from Example 1.

[0029] Figure 3 This is the blood drug concentration-time curve of Example 1. Detailed Implementation

[0030] To enhance understanding of the present invention, the present invention will be further described in detail below with reference to embodiments and accompanying drawings. These embodiments are only used to explain the present invention and do not constitute a limitation on the scope of protection of the present invention.

[0031] Example 1: Formulation and preparation of galangin nanocrystal suspension (1)

[0032] Prescription composition:

[0033]

[0034] Preparation method: Gingerol was weighed and added to anhydrous ethanol, and stirred until completely dissolved to obtain an organic phase; stabilizer poloxamer 407 and polyvinylpyrrolidone K30 were weighed and added to purified water, and stirred to dissolve to obtain an aqueous phase; the aqueous phase was placed in a high-speed shear machine at 800 rpm, and the organic phase was slowly and uniformly injected into the aqueous phase, and sheared continuously for 15 min to obtain a coarse suspension; the coarse suspension was placed in a high-pressure homogenizer at 1200 bar and circulated 10 times; ethanol was removed by rotary evaporation under reduced pressure at 40℃, and the suspension was filtered through a 0.22 μm filter membrane to obtain a galangin nanocrystal suspension with a particle size of up to 122.2 nm.

[0035] Example 2: Formulation and preparation of galangin nanocrystal suspension (2)

[0036] Prescription composition:

[0037]

[0038] Preparation method: Galangin was weighed and added to anhydrous ethanol, stirred until completely dissolved to obtain the organic phase; hydroxypropyl methylcellulose, the stabilizer, was weighed and added to purified water, stirred and dissolved to obtain the aqueous phase; the aqueous phase was placed in a high-speed shear mill at 800 rpm, and the organic phase was slowly and uniformly injected into the aqueous phase, and sheared continuously for 15 min to obtain a coarse suspension; the coarse suspension was placed in a high-pressure homogenizer at 1200 bar and circulated 10 times; ethanol was removed by rotary evaporation under reduced pressure at 40℃, and the solution was filtered through a 0.22 μm filter membrane to obtain a galangin nanocrystal suspension with a particle size of up to 242.2 nm. In this embodiment, only hydroxypropyl methylcellulose was used as a stabilizer, and the high viscosity of the solution system limited the efficiency of high-pressure homogenization, resulting in a larger particle size and wider distribution.

[0039] Example 3: Formulation and preparation of galangin nanocrystal suspension (3)

[0040] Prescription composition:

[0041]

[0042] The preparation method is the same as in Example 1. The stabilizer contains only poloxamer 407 and the weight ratio of it to galangin is 1:1. The particle size of the galangin nanocrystal suspension prepared using this formula can reach 168.1 nm.

[0043] Example 4: Preparation of galangin nanocrystal suspension (4)

[0044] Prescription composition:

[0045]

[0046] The preparation method is the same as in Example 1. The stabilizer contains only polyvinylpyrrolidone K30, and the weight ratio of it to galangin is 1:1. The particle size of the galangin nanocrystal suspension prepared using this formula can reach 176.7 nm.

[0047] Example 5: Preparation of galangin nanocrystal suspension (5)

[0048] Prescription composition:

[0049]

[0050] The preparation method is the same as in Example 1. The stabilizers are poloxamer 407 and polyvinylpyrrolidone K30. The weight ratio of stabilizer to galangin is 1:2. The particle size of the galangin nanocrystal suspension prepared using this formula can reach 130.2 nm.

[0051] Example 6: Preparation of galangin nanocrystal suspension (6)

[0052] Prescription composition:

[0053]

[0054] The preparation method is the same as in Example 1. The stabilizers are poloxamer 407 and polyvinylpyrrolidone K30. The weight ratio of stabilizer to galangin is 2:1. The particle size of the galangin nanocrystal suspension prepared using this formula can reach 145.4 nm.

[0055] Example 7: Preparation of galangin nanocrystal suspension (7)

[0056] Prescription composition:

[0057]

[0058] Preparation method: Gingerol was weighed and added to anhydrous ethanol, and stirred until completely dissolved to obtain an organic phase; poloxamer 407 and polyvinylpyrrolidone K30 were weighed and added to purified water, and stirred to dissolve to obtain an aqueous phase; the aqueous phase was placed in a high-speed shearing machine at 800 rpm, and the organic phase was slowly and uniformly injected into the aqueous phase, and sheared continuously for 15 min to obtain a coarse suspension; the coarse suspension was placed in a probe sonicator and sonicated at 300W for 10 min; ethanol was removed by rotary evaporation under reduced pressure at 40℃, and the suspension was filtered through a 0.22 μm filter membrane to obtain a galangin nanocrystal suspension with a particle size of up to 190.3 nm.

[0059] Example 8: Preparation of galangin nanocrystal suspension (8)

[0060] The prescription and preparation method are the same as in Example 1, except that the high-pressure homogenization pressure of 1200 bar is changed to 1000 bar.

[0061] Example 9: Preparation of galangin nanocrystal suspension (9)

[0062] The prescription and preparation method are the same as in Example 1, except that the high-pressure homogenization pressure of 1200 bar is changed to 1500 bar.

[0063] Example 10: Preparation of galangin nanocrystal suspension (10)

[0064] The prescription and preparation method are the same as in Example 1, except that the number of high-pressure homogenization cycles is changed from 10 to 5.

[0065] Example 11: Preparation of galangin nanocrystal suspension (11)

[0066] The prescription and preparation method are the same as in Example 1, except that the number of high-pressure homogenization cycles is changed from 10 to 15.

[0067] The particle size and PDI of the galangin nanocrystal suspensions prepared in Examples 1-11 were measured using particle size and PDI as evaluation indicators. The results are shown in the table below:

[0068]

[0069] In summary, as shown in the table, the particle size of the galangin nanocrystal suspension is between 100 and 200 nm. The suspensions obtained using a single stabilizer in Examples 2, 3, and 4 have a PDI greater than 0.3. In Example 2, hydroxypropyl methylcellulose was used as a stabilizer, which may have resulted in a larger particle size and wider distribution due to the higher viscosity of the solution system, limiting the high-pressure homogenization efficiency. The suspension obtained by the antisolvent precipitation-high-pressure homogenization method in Example 1 has a PDI less than 0.3 and a narrower particle size distribution. Examples 5 and 6 improved the stability by adjusting the weight of the stabilizer and galangin. The proportion of PDI is between 0.2 and 0.3; the ultrasonic process used in Example 7 resulted in a PDI greater than 0.3; from the homogenization pressures of 1000 bar and 1500 bar in Examples 8 and 9, compared with the particle size of Example 1, further increasing the pressure did not significantly improve the particle size, and the PDI did not change significantly. Therefore, a homogenization pressure of 1200 bar is optimal; from the comparison of Examples 10 and 11 with Example 1, the target particle size can be achieved after 10 homogenization cycles, and further cycles do not provide significant optimization.

[0070] Example 12 Stability Experiment

[0071] The galangin nanocrystal suspensions obtained in Examples 1, 5, and 6 were placed at room temperature for 3, 7, and 15 days, respectively. The particle size and PDI were measured and compared with the results of day 0. The results are shown in the table below:

[0072]

[0073] The results showed that the suspension prepared in Example 1 was the most stable. Too low a stabilizer ratio would easily lead to agglomeration, while too high a ratio would increase the particle size. If the stabilizer dosage was too low, a complete protective layer could not be formed on the surface of the nanocrystals, and the particles would easily aggregate and settle, resulting in poor stability. If the dosage was too high, it would lead to increased system viscosity and agglomeration of interparticle bridging, which would also compromise the stability of the system.

[0074] Example 13: The characterization results of the galangin nanocrystals prepared in Example 1 are as follows:

[0075] like Figure 1 As shown, in the preferred embodiment, the nanocrystals are spherical, uniformly dispersed, and have a particle size of 100~150nm.

[0076] To better investigate the bioavailability of galangin nanocrystals, in vitro release and in vivo pharmacokinetic studies were conducted on the galangin nanocrystals prepared in Example 1. Galangin raw material was used as a control; the raw material was a slightly yellow needle-like crystal or a pale yellow needle-like crystalline powder, manufactured by Chengdu Glip Biotechnology Co., Ltd.

[0077] (1) In vitro release study

[0078] An appropriate amount of the active pharmaceutical ingredient and the galangin nanocrystal suspension from Example 1 were transferred to a dialysis bag with a molecular weight cutoff of 3500 Da. The dialysis bag was sealed tightly at both ends and placed in 100 mL of pH 6.8 phosphate buffer (containing 1% Tween 80). The solution was shaken at 100 rpm at 37°C. The release medium was collected at different time points, and an equal amount of fresh medium was added promptly. The collected solution was filtered through a membrane and analyzed. The results are as follows: Figure 2 As shown.

[0079] The results showed that the preferred embodiment achieved a cumulative dissolution rate of 60% at 60 min and reached equilibrium at 8 h, with a cumulative dissolution rate as high as 99%; while the active pharmaceutical ingredient (API) only had a cumulative in vitro release of about 75% after 48 h. Compared with the API, the nanocrystals had a significant solubilizing effect.

[0080] (2) Pharmacokinetic study in vivo

[0081] SD rats were randomly divided into two groups of six each. They were fasted for 12 hours before administration but allowed free access to water. The active pharmaceutical ingredient (API) and galangin nanocrystal suspension (1) were administered intravenously (dose: 0.5 mg / kg). Blood samples were collected from the orbital cavity at different time points after administration. The blood samples were centrifuged at 3000 rpm for 15 min, and plasma was collected for analysis. Pharmacokinetic parameters were calculated using PKSolver 2.0, and the results are as follows: Figure 3 As shown.

[0082] The results showed that the t of galangin nanocrystal suspension 1 / 2 The increased AUC and decreased CL and Vd indicate that nanocrystals can slow down the elimination rate of drugs in vivo, prolong the duration of action, and thus improve bioavailability.

[0083] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A galangin nanocrystal suspension, characterized in that, It contains galangin as an active ingredient, a stabilizer, and water, wherein the average particle size of the galangin is 100-150 nm.

2. The galangin nanocrystal suspension according to claim 1, characterized in that, The stabilizer is selected from one or more of poloxamer 407, polyvinylpyrrolidone K30, hydroxypropyl methylcellulose, poloxamer 188, and vitamin E polyethylene glycol succinate.

3. The galangin nanocrystal suspension according to claim 1, characterized in that, The mass ratio of galangin to stabilizer is 1:0.5 to 1:

2.

4. The galangin nanocrystal suspension according to claim 2, characterized in that, The mass ratio of galangin to stabilizer is 1:

1.

5. The galangin nanocrystal suspension according to claim 4, characterized in that, The stabilizer is a compound stabilizer composed of Loxamer 407 and polyvinylpyrrolidone K30 in a mass ratio of 1:

1.

6. The galangin nanocrystal suspension according to claim 5, characterized in that, The method for preparing the suspension includes the steps of preparing a galangin nanocrystal suspension by using antisolvent precipitation-ultrasound method or antisolvent precipitation-high pressure homogenization method with galangin, stabilizer and water.

7. The galangin nanocrystalline pharmaceutical composition according to claim 6, characterized in that, The suspension is prepared by antisolvent precipitation-high pressure homogenization, specifically including the following steps: (1) Alpinin was dissolved in an organic solvent to prepare the organic phase of the drug; (2) Dissolve the stabilizer in purified water to obtain an aqueous phase; (3) The organic phase was injected into the aqueous phase under high-speed shearing conditions to obtain a crude suspension; (4) The coarse suspension was placed in a high-pressure homogenizer for circulation homogenization; (5) Remove the organic solvent and filter to obtain galangin nanocrystal suspension.

8. The galangin nanocrystalline pharmaceutical composition according to claim 7, characterized in that, The organic solvent was anhydrous ethanol, the concentration of galangin in the organic phase was 10 mg / mL, and the volume ratio of the aqueous phase to the organic phase was 8:

1.

9. The galangin nanocrystalline pharmaceutical composition according to claim 7, characterized in that, In step 3), the high-speed shearing speed is 800 rpm, and in step 4), the high-pressure homogenization pressure is 1000~1500 bar, and the number of cycles is 5~15.

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