A betamethasone dipropionate composition and nano-lyophilized agent thereof and a preparation method thereof

Abstract

The present application provides a betamethasone dipropionate composition and nanocrystalline lyophilizate thereof and a preparation method. The betamethasone dipropionate composition is composed of betamethasone dipropionate, specific surfactants and high molecular polymers. After being mixed with water to form a suspension, the wet medium grinding is adopted to obtain a nanosuspension, and the nanocrystalline lyophilizate of betamethasone dipropionate is prepared by freeze-drying technology. The nanocrystalline lyophilizate has very high saturated solubility, shows excellent effect in the experiment on a psoriasis mouse model, and has simple and controllable production process, is green and environment-friendly, and is suitable for industrial production.

Description

Technical Field

[0001] This invention relates to a betamethasone dipropionate composition and its nano-lyophilized formulation, as well as its preparation method. It belongs to the field of pharmaceutical formulation technology. Background Technology

[0002] Psoriasis is a chronic inflammatory skin condition characterized by erythematous plaques and silvery-white scales. Epidemiological surveys indicate an incidence rate of up to 2%. For example, in the United States, approximately 2% to 3% of adults are affected by psoriasis. It is estimated that 55.6% of patients are classified as mild plaque psoriasis, 37.6% as moderate, and 6.6% as severe. Although psoriasis can affect the whole body, the scalp and extremities (such as elbows and knees) are the most commonly affected. The scalp is often one of the first areas affected in psoriasis patients, and the longer a patient has had psoriasis, the greater the likelihood of scalp lesions developing. The skin is an ideal target for topical treatments, but it is also a barrier to the effective penetration and absorption of drugs. The excessive proliferation of keratinocytes associated with psoriasis further exacerbates this barrier. Although oral and injectable treatments overcome the need to penetrate the stratum corneum, the associated risks of infection and liver, kidney, and bone marrow toxicity may limit their use in some patients. Most patients with mild to moderate psoriasis receive topical treatments, with corticosteroids being the most common.

[0003] Betamethasone dipropionate (BD) is a fifth-generation corticosteroid that inhibits phospholipase activity, thereby inhibiting the synthesis of arachidonic acid, prostaglandins, and leukotrienes, achieving immunosuppression, anti-inflammation, and anti-proliferation effects. In clinical practice, BD is widely used to treat various skin diseases. Currently, BD is mainly available in ointments, creams, and lotions. BD belongs to the category of insoluble drugs, and insoluble drugs often present a challenging problem in drug development. Reports indicate that when particles are small enough to be nanoscale, they can increase the dissolution rate of compounds, ensuring close contact with the stratum corneum of the skin, thus promoting the penetration of active compounds into the skin. Therefore, some literature reports the development of BD-loaded nanoliposome carriers for the treatment of dermatitis, psoriasis, and inflammation; however, this method requires a large amount of carrier material, and the process steps are cumbersome, posing certain difficulties for industrialization.

[0004] Patent application number 2010101754347 discloses a method for preparing betamethasone dipropionate microparticles. The method involves dissolving betamethasone dipropionate in a solvent, then adding it to an antisolvent under stirring, and continuing stirring to induce crystallization. However, the resulting microparticles have a particle size distribution mainly between 5-10 μm, indicating poor microparticle formation and inconsistent uniformity. Patent application number 202110301592 discloses a method for preparing betamethasone dipropionate nanoparticles, also using an antisolvent method, yielding a product with a particle size D50 of approximately 500 nm. However, antisolvent methods utilize large amounts of organic solvents, easily leading to a high risk of residual solvent in the product. Furthermore, the use of large amounts of organic solvents in production lacks safety and environmental friendliness, making it unsuitable for industrial production. Summary of the Invention

[0005] The purpose of this invention is to provide a betamethasone dipropionate nano-lyophilized formulation with small particle size and high saturation solubility. Another purpose of this invention is to provide the use of this nano-formulation as a topical skin medication for the treatment of psoriasis.

[0006] This invention unexpectedly reveals that a stable betamethasone dipropionate nanoformulation can be prepared from a combination of betamethasone dipropionate, a specific surfactant, and a polymer. In particular, the prepared betamethasone dipropionate lyophilized powder, after reconstitution, has a particle size D90 of less than 1000 nm, falling within the nanoscale range. This effectively improves the saturated solubility of betamethasone dipropionate and enhances its bioavailability. Furthermore, the preparation method of this invention is simple, environmentally friendly, and the process is easy to control, making it suitable for large-scale industrial production.

[0007] The technical solution of the present invention is as follows:

[0008] The invention provides a betamethasone dipropionate composition, which, by weight percentage, comprises 20% to 75% betamethasone dipropionate, 12.5% ​​to 40% surfactant, and 12.5% ​​to 40% polymer, characterized in that the surfactant is dodecyl sulfonate and the polymer is polyvinyl ketone.

[0009] Preferably, in the above-described betamethasone dipropionate composition, the surfactant is sodium dodecyl sulfonate and the polymer is povidone K30 (PVP K30).

[0010] As another objective of this invention, the application of the above-described betamethasone dipropionate composition in the preparation of betamethasone dipropionate nanoformulations is also provided.

[0011] Preferably, in the above-described application, the betamethasone dipropionate nanoformulation is a betamethasone dipropionate nano-lyophilized agent.

[0012] As another objective of the present invention, a betamethasone dipropionate nano-lyophilized agent is provided, characterized in that it is made from the betamethasone dipropionate composition described above and water.

[0013] Preferably, in the above-mentioned betamethasone dipropionate nano-lyophilized agent, the weight ratio of betamethasone dipropionate to water is 1:100 to 1:2000, more preferably 1:500 to 1:1000.

[0014] As another objective of this invention, a method for preparing the above-described betamethasone dipropionate nano-lyophilized agent is provided, characterized by comprising the following steps:

[0015] Step 1: Prepare a suspension by mixing betamethasone dipropionate, surfactant, polymer and water in the specified proportions;

[0016] Step 2: Pre-treat the suspension on a homogenizer at a speed of not less than 6000 rpm;

[0017] Step 3: Grind the pretreated suspension.

[0018] Step 4: Freeze-dry the ground suspension to solidify it.

[0019] Furthermore, in step 2, the pretreatment is carried out on a homogenizer at a speed of not less than 6000 rpm, preferably 8000-10000 rpm. The purpose is to break up the large pieces of betamethasone dipropionate in advance, so that the particle size of the raw material in the suspension entering the ball mill is more uniform, thereby ensuring smoother grinding in the next step of the ball mill.

[0020] Preferably, in the method described above, a ball mill is used for media grinding in step 3; preferably, the ball mill chamber is filled with zirconium oxide beads.

[0021] Furthermore, in step 3, the parameters for media grinding using a ball mill include: the diameter and quantity of the zirconia beads used for grinding, the rotation speed of the mill, and the speed of the peristaltic pump used for circulation.

[0022] Furthermore, in step 3, the diameter of the zirconia beads is 0.2–0.8 mm, preferably 0.3–0.6 mm; the amount of zirconia beads used is 30%–80% of the volume of the ball mill cavity.

[0023] Furthermore, in step 3, the rotation speed of the grinding mill is 1000-3500 rpm, preferably 2000-3000 rpm.

[0024] Furthermore, in step 3, the speed of the peristaltic pump is 100-300 rpm, preferably 150-250 rpm.

[0025] Furthermore, in step 3, the grinding time is 30 to 300 minutes, preferably 60 to 180 minutes.

[0026] Preferably, in step 4, the temperature of the pre-freezing stage is -30 to -40°C, and the time is 2 to 4 hours; the temperature of the first drying stage is -10 to 0°C, and the time is 16 to 24 hours, with a vacuum degree of less than 0.3 mbar; the temperature of the second drying stage is 0 to 25°C, and the time is 5 to 8 hours, with a vacuum degree of less than 0.6 mbar.

[0027] As another object of the present invention, it is also provided that the above-described betamethasone dipropionate nano-lyophilized agent is used in the preparation of a topical skin medicine for treating psoriasis. In use, the betamethasone dipropionate nano-lyophilized agent needs to be reconstituted with purified water before use, and the concentration can be prepared as needed.

[0028] This invention unexpectedly reveals that a stable betamethasone dipropionate nano-formulation can be prepared from a combination of betamethasone dipropionate, a specific surfactant, and a polymer. In particular, the betamethasone dipropionate lyophilized powder prepared by this invention exhibits a particle size (D90) of less than 1000 nm after reconstitution, falling within the nanoscale range. This effectively improves the saturated solubility of betamethasone dipropionate and enhances its bioavailability. The betamethasone dipropionate nano-lyophilized formulation prepared by this invention showed no significant change in particle size after long-term stability testing, demonstrating excellent sample stability. Furthermore, the preparation method of this invention is simple, environmentally friendly, and easy to control, making it suitable for large-scale industrial production.

[0029] Compared with the prior art, the present invention has the following significant advantages: (1) It effectively reduces the average particle size of the product, and the prepared betamethasone dipropionate lyophilized powder is nano-sized (D90 < 500 nm), and the particle size D90 after reconstitution is also less than 1000 nm; (2) By freeze-drying the betamethasone dipropionate nano suspension, the degradation reaction of betamethasone dipropionate is effectively eliminated, the proportion of active drug components is greatly increased, and it is convenient for storage and transportation, fully meeting the needs of storage and treatment; (3) By reducing the particle size of betamethasone dipropionate to the nano-sized, the saturated solubility can be effectively improved and the bioavailability can be increased; (4) The pharmacodynamic evaluation conducted on psoriasis model mice showed a very significant therapeutic effect; (5) The nano-formulation has a simple prescription, the process is green and environmentally friendly, and it is suitable for industrial production. Attached Figure Description

[0030] Figure 1 Comparative Pharmacodynamic Study in a Mouse Model of Psoriasis – Simplified PASI Score

[0031] In the figure, the horizontal axis "Day" represents the time unit "day"; the vertical axis "Erythema score" represents the "erythema score"; and the vertical axis "Scaly score" represents the "skin lesion score". Significant differences were observed compared to the control group (*P < 0.05); significant differences were observed compared to the model group (#P < 0.05); and significant differences were observed between groups (&P < 0.05).

[0032] Figure 2 Comparative pharmacodynamic study in a mouse model of psoriasis—dermatopathology HE staining

[0033] The scale bar in the figure is 100 μm. Detailed Implementation

[0034] The following examples help to understand the present invention, but do not limit the scope of the invention. All raw materials and reagents used in the embodiments of the present invention are commercially available.

[0035] Example 1: Preparation of Betamethasone Dipropionate Nano-Lyophilized Agent

[0036]

[0037] Preparation process:

[0038] (1) Add betamethasone dipropionate, sodium dodecyl sulfonate, PVP K30 and purified water to a beaker and stir to obtain a suspension for later use.

[0039] (2) The above suspension was pretreated for 10 minutes using an IKA homogenizer at a speed of 8000 rpm.

[0040] (3) Add the pretreated suspension to the storage tank of the ball mill. The ball mill chamber contains 100 ml of 0.4 mm zirconium oxide beads. Set the peristaltic pump speed to 200 rpm and the ball mill speed to 3000 rpm. Time for 60 min and collect the nano suspension.

[0041] (4) Pour the collected nano-suspension into a stainless steel tray and freeze-dry it in a freeze dryer. Parameters: In the freezing stage, the sample was frozen at -40℃ for 3 hours. In the first drying stage, the sample was frozen at -10℃ for 6 hours, then at -5℃ for 6 hours, and then at 0℃ for 4 hours, all under a vacuum of 0.3 mbar. In the second drying stage, the temperature was raised to 25℃ and held under a vacuum of 0.06 mbar for 5 hours.

[0042] (5) Collect solid materials.

[0043] Example 2: Preparation of Betamethasone Dipropionate Nano-Lyophilized Agent

[0044]

[0045]

[0046] Preparation process: Same as in Example 1.

[0047] Example 3: Preparation of Betamethasone Dipropionate Nano-Lyophilized Agent

[0048]

[0049] Preparation process: Same as in Example 1.

[0050] Example 4: Preparation of Betamethasone Dipropionate Nano-Lyophilized Agent

[0051]

[0052] Preparation process: In step (3), 90 ml of 0.3 mm zirconia beads were placed in the ball mill chamber, the peristaltic pump speed was set to 200 rpm, the ball mill speed was set to 2000 rpm, and the time was set to 120 min to collect the nano suspension. The remaining steps are the same as in Example 1.

[0053] Example 5: Preparation of Betamethasone Dipropionate Nano-Lyophilized Agent

[0054]

[0055] Preparation process: In step (3), 110 ml of 0.2 mm zirconia beads were placed in the ball mill chamber, the peristaltic pump speed was set to 300 rpm, the ball mill speed was set to 2500 rpm, and the time was set to 120 min to collect the nano suspension. The remaining steps are the same as in Example 1.

[0056] Comparative Example 1: Preparation of Betamethasone Dipropionate Lyophilized Formulation

[0057]

[0058] Preparation process: Same as in Example 1

[0059] Comparative Example 2: Preparation of Betamethasone Dipropionate Lyophilized Formulation

[0060]

[0061] Preparation process: Same as in Example 1

[0062] Particle size determination of betamethasone dipropionate lyophilized formulation composition

[0063] Particle size was measured using Examples 1-5 and Comparative Examples 1-2 based on photon correlation spectroscopy, performed by a Brookhaven Instruments 90Plus PALS (Brookhaven, USA) equipped with a 40mW diode laser (640nm). Prior to measurement, the samples were diluted or redispersed with deionized water to a BD concentration of approximately 0.05 mg / mL. Specific results are shown in Table 1 below.

[0064] Table 1. Particle size results of Examples 1-5 and Comparative Examples 1-2

[0065]

[0066] As can be seen from the table, after resolidation, the particle size D90 of the samples in Examples 1 to 5 is less than 1000 nm, reaching the nanoscale range.

[0067] Stability study of betamethasone dipropionate nanocrystalline lyophilized formulation composition

[0068] The samples in Example 3 underwent long-term stability testing according to the guidelines for drug stability testing in Appendix II of the 2015 edition of the Chinese Pharmacopoeia to examine whether the particle size would change during storage.

[0069] Long-term test: Samples were sealed and placed at 25℃ and RH 60% ± 10%, and particle size was measured at 3, 6, 9, and 12 months. Before measurement, the samples were diluted or redispersed with deionized water to a BD concentration of approximately 0.05 mg / mL. The measurement results are shown in Table 2 below.

[0070] Table 2. Long-term stability particle size results of Example 3

[0071]

[0072] The results above show that the particle size of the betamethasone dipropionate nanocrystal lyophilized formulation in Example 3 did not change significantly after long-term stability testing, indicating good sample stability.

[0073] Comparative Study on Saturated Solubility Determination

[0074] The purpose of this experiment is to study the differences in saturated solubility of nano-formulations, non-nano-formulations, and pure BD active pharmaceutical ingredient.

[0075] Comparative example: Preparation of non-nano formulations

[0076]

[0077] Preparation process:

[0078] (1) Add betamethasone dipropionate, sodium dodecyl sulfonate, PVP K30 and purified water into a beaker and stir to obtain a suspension for later use.

[0079] (2) The above suspension was pretreated for 10 minutes using an IKA homogenizer at a speed of 8000 rpm.

[0080] (3) Pour the pretreated suspension into a stainless steel tray and freeze-dry it in a freeze dryer. Parameters: In the freezing stage, freeze the sample at -40℃ for 3 hours. In the first drying stage, freeze the sample at -10℃ for 6 hours, then at -5℃ for 6 hours, and then at 0℃ for 4 hours, all under a vacuum of 0.3 mbar. In the second drying stage, raise the temperature to 25℃ and maintain it under a vacuum of 0.06 mbar for 5 hours.

[0081] (4) Collect solid materials.

[0082] Particle size determination: The determination method is the same as above. Before determination, the sample was diluted or redispersed with deionized water to a BD concentration of approximately 0.05 mg / mL. Results: D10 was 880 nm; D50 was 3070 nm; D90 was 16430 nm.

[0083] The saturated solubility of nano-formulation (Example 3), non-nano-formulation (Comparative Example), and BD active pharmaceutical ingredient was determined in purified water. A sample equivalent to 4 mg of BD was dispersed in 5 mL of purified water and stirred at 75 rpm in a water bath at 25°C. After 24 hours, the suspension was transferred to a centrifuge tube and centrifuged at 16000 × g for 15 minutes. The supernatant was analyzed by HPLC. HPLC method: mobile phase acetonitrile:water = 44:56; column: octadecylsilane-bonded silica column 4.6*250 mm 5 μm; monitoring wavelength: 254 nm; flow rate: 1 mL / min; column temperature: 30°C; injection volume: 10 μL; sample concentration: 300 μg / mL.

[0084] The saturated solubility results are shown in Table 3. The results in the table show that the solubility of Example 3 was significantly improved, being 202 times and 57 times that of the BD raw material and the comparative example, respectively.

[0085] Table 3. Saturated solubility results

[0086] Nanoparticle formulation (Example 3) 92.9 Non-nano formulations (comparative example) 1.63 BD raw materials 0.46

[0087] Comparative pharmacodynamic studies in mouse models of psoriasis

[0088] The purpose of this experiment was to compare the efficacy differences between nano-formulations, non-nano-formulations, and methotrexate (MTX, a drug used to treat psoriasis) in a mouse model of psoriasis.

[0089] Materials and methods

[0090] Thirty adult BALB / c mice (SYXK(Su)2018-0008) were randomly divided into four groups: a normal control group (Control), an imiquimod model group (Model), a methotrexate positive drug group (MTX), a non-nanoformation group (Comparative), and a nanoformation group (Example 3), with six mice in each group. Mice in the Model, MTX, and treatment groups were treated with 62.5 mg of 5% imiquimod (IMQ) cream on the shaved area of ​​their backs, while mice in the control group were treated with 62.5 mg of petroleum jelly, once daily for 14 consecutive days. All experimental procedures were performed in accordance with the requirements of the Animal Ethics Committee (Ethics No.: XMLL-2022-051).

[0091] After successful modeling, mice in the MTX group were given 0.4 mL of methotrexate suspension (0.023 mg / mL) by gavage; mice in the non-nano formulation group (comparative example) and the betamethasone dipropionate nanocrystal group (Example 3) were given 1 mL of drug suspension (0.02 mg / mL) of their respective concentrations on the affected areas of the mice.

[0092] A simplified PASI score was used to assess the degree of erythema and scaling in mouse skin, ranging from 0 to 4 points. The results showed that, compared to the model group, MTX, the comparative case, and Example 3 all reduced the degree of erythema and scaling in mouse skin, with Example 3 showing the best effect (see [link to example]). Figure 1 ).

[0093] HE staining of the skin pathology showed that the Model group mice had significant epidermal hyperplasia with hyperkeratosis, significantly thickened stratum spinosum, elongated epidermal ridges, and numerous lymphocytic infiltrations in the dermis, exhibiting typical psoriatic-like skin pathological changes. In the MTX group, the comparative group, and the Example 3 group mice, the thickness of the epidermal layer, hyperkeratosis, parakeratosis, acanthosis, and the degree of lymphocytic infiltration were all reduced compared to the Model group, with the Example 3 group showing the most significant improvement in skin lesion histopathology (see Example 3). Figure 2 ).

Claims

1. A betamethasone dipropionate nano-lyophilized agent, characterized in that... It is prepared by a method comprising the following steps: a betamethasone dipropionate composition and water, wherein the betamethasone dipropionate composition comprises, by weight percentage, 20%~75% betamethasone dipropionate, 12.5%~40% surfactant, and 12.5%~40% polymer, wherein the surfactant is sodium dodecyl sulfonate, and the polymer is povidone K30; Step 1, preparing a suspension by mixing betamethasone dipropionate, surfactant, polymer, and water in a specified ratio; Step 2, pretreating the suspension on a homogenizer at a speed of not less than 6000 rpm; Step 3, grinding the pretreated suspension; Step 4, freeze-drying the ground suspension.

2. The betamethasone dipropionate nano-lyophilized agent according to claim 1, wherein the weight ratio of betamethasone dipropionate to water is 1:100 to 1:2000.

3. A method for preparing the betamethasone dipropionate nano-lyophilized agent according to claim 1 or 2, characterized in that... Includes the following steps: Step 1: Prepare a suspension by mixing betamethasone dipropionate, surfactant, polymer and water in the specified proportions; Step 2: Pre-treat the suspension on a homogenizer at a speed of not less than 6000 rpm; Step 3: Grind the pretreated suspension; Step 4: Freeze-dry the ground suspension to solidify it.

4. The method according to claim 3, wherein a ball mill is used for media grinding in step 3.

5. The method according to claim 4, characterized in that... The ball mill chamber is filled with zirconium oxide beads.

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