High bioavailability albendazole nanocrystals and methods of making the same

Abstract

The application discloses a kind of high bioavailability albendazole nanocrystals and preparation method thereof.The nanocrystals of the application are prepared by mixing albendazole, natural source glycoside stabilizer and water to form a suspension, and then using wet medium grinding.The average particle size is 100-500nm.The nanocrystals are obtained by dispersing albendazole in natural source glycoside solution and then nano-treating, with high drug loading, which can greatly improve the solubility and dissolution of albendazole, and then improve its absorption and bioavailability.The stabilizer used in the application is abundant in resources, natural and economical, and the preparation process is simple, efficient and safe, which is suitable for industrial production, and provides a high bioavailability intermediate for albendazole preparation.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a highly bioavailable albendazole nanocrystal and its preparation method. Background Technology

[0002] Albendazole, chemically named methyl 5-(propylthio)-2-benzimidazole carbamate, has the molecular formula C2. 12 H 15 N3O2S, molecular weight: 265.33. Albendazole is a first-line broad-spectrum anthelmintic that is effective in treating intestinal and systemic parasitic diseases. However, this drug belongs to BCS Class II drugs, has poor water solubility, low absorption in the intestines, low distribution in plasma and liver tissues, and low bioavailability. Long-term use at high doses can easily lead to adverse drug reactions. Therefore, it is necessary to adopt appropriate methods to improve its solubility, increase its absorption in vivo, and improve its bioavailability.

[0003] Nanocrystals are submicron colloidal dispersion systems composed of only drugs and a small amount of stabilizers. They have high drug loading capacity, simple preparation process, and can achieve targeted targeting of lesion sites through surface modification. They can significantly improve the dissolution and bioavailability of poorly soluble drugs, and are an important strategy to improve the success rate of new drug development, reduce adverse reactions, and improve drug efficacy.

[0004] The preparation methods of nanocrystals are divided into top-down and bottom-up methods. The top-down method involves reducing the size of solid drug particles to the nanoscale through high-pressure homogenization or nano-milling; the bottom-up method involves dissolving the drug in a solvent, then adding an antisolvent to precipitate it, and finally obtaining nanoscale powder by spraying or freeze-drying. Wet ball milling has the advantages of mature technology, simple operation, wide applicability, narrow particle size distribution of the prepared nanomedicines, and no organic solvent residue, and has become the most commonly used preparation method for commercially available nanomedicines (Malamatari M, Taylor K MG, Malamatari S, et al. Pharmaceutical nanocrystals: production by wet milling and applications[J]. Drug Discov Today,2018,23(3):534-547).

[0005] To improve the solubility of albendazole, researchers have successively developed and studied different formulations such as solid dispersions, inclusion complexes, and liposomes. However, these formulations have problems such as limited ability to improve the bioavailability of albendazole, excessive excipient loading, and difficulties in industrialization.

[0006] Currently, research on albendazole nanocrystals often uses synthetic polymers such as poloxamer 188, PVP K30, and Tween 80 as stabilizers, and employs high-pressure homogenization or anti-solvent precipitation methods for preparation. However, these methods suffer from problems such as high energy consumption, the use of organic solvents, and the large amount of stabilizers required. Summary of the Invention

[0007] Objective of this invention: To address the shortcomings of existing technologies, this invention provides albendazole nanocrystals with high bioavailability and a method for their preparation. This invention improves the bioavailability of albendazole by screening natural-derived glycoside stabilizers and optimizing the wet media milling process. The preparation method is simple, easy to scale up for production, and the prepared albendazole nanocrystals effectively exert the antiparasitic therapeutic effect of albendazole.

[0008] Technical solution: The objective of this invention is achieved through the following technical solution:

[0009] This invention provides albendazole nanocrystals with high bioavailability. The nanocrystals are prepared by mixing albendazole, a naturally derived glycoside stabilizer and water to form a suspension, and then preparing the suspension by wet grinding. The average particle size is 100-500 nm.

[0010] This invention utilizes a naturally derived glycoside stabilizer mixed with albendazole to prepare albendazole nanocrystals, fully leveraging the stabilizer's advantages: 1) coating the albendazole particle surface with the stabilizer through physical adsorption or hydrogen bonding, increasing steric hindrance and preventing drug particle aggregation; 2) maintaining the drug molecules in a supersaturated state for a certain period; and 3) increasing drug solubility. Experiments show that the saturated solubility of albendazole nanocrystals in water, pH 1.0 hydrochloric acid solution, pH 4.5 acetate buffer, and pH 6.8 phosphate buffer is approximately 3 to 2500 times higher than that of albendazole.

[0011] In a preferred embodiment of the present invention, the albendazole nanocrystals have an average particle size of 100–300 nm and a zeta potential range of -30–-20 mV.

[0012] Preferably, the natural-derived stabilizer is selected from tea saponin or sucralose.

[0013] More preferably, the naturally sourced glycoside stabilizer is selected from tea saponin. Using tea saponin as a stabilizer is widely available, inexpensive, readily available, and safe.

[0014] Preferably, the mass ratio of albendazole to the natural glycoside stabilizer is 10:1 to 10:12; and the concentration of albendazole in the suspension is 0.2% to 3.0%.

[0015] More preferably, the mass ratio of albendazole to the natural glycoside stabilizer is 5:1 to 5:4; and the concentration of albendazole in the suspension is 0.5% to 2.5%.

[0016] This invention optimizes the type and dosage of stabilizers and the wet media milling process parameters, thereby improving the solubility and bioavailability of the poorly soluble drug albendazole.

[0017] This invention also provides a method for preparing the highly bioavailable albendazole nanocrystals, comprising the following steps:

[0018] (1) Thoroughly mix the natural glycoside stabilizer with water to form stabilizer solution A;

[0019] (2) Add albendazole to the stabilizer solution A obtained in step (1), stir to suspend the drug in it, and obtain albendazole crude suspension B;

[0020] (3) The albendazole crude suspension B obtained in step (2) is subjected to wet media milling to obtain albendazole nano suspension, namely the albendazole nanocrystals.

[0021] This invention employs nano-grinding technology to treat albendazole. During the grinding process, the drug and stabilizer are co-ground, which effectively prevents drug particle aggregation and ensures the stability of the suspension. Stability was assessed for 28 days at both room temperature and low temperature; no significant changes were observed in the nanocrystal particle size and PDI.

[0022] Preferably, in step (3), the process parameters for wet media grinding are: grinding time 2-8h, grinding speed controlled at 300-700rpm, and liquid droplet ratio 1:(2-8).

[0023] More preferably, the process parameters for the wet media grinding are: grinding time 3-7 hours, grinding speed controlled at 400-700 rpm, and liquid droplet ratio 1:(4-8).

[0024] Preferably, in step (3), the grinding beads used in the wet media grinding have a particle size of 0.2 to 0.5 mm and are made of zirconium oxide, yttrium-stabilized zirconium oxide, or stainless steel.

[0025] The present invention also provides an albendazole nanocrystal freeze-dried solid powder, which is obtained by freeze-drying the aforementioned albendazole nanocrystals.

[0026] Preferably, the freeze-drying protectant used in the freeze-drying is selected from any one or a combination of sucrose, lactose, fructose, mannitol, sorbitol, xylitol, and polyethylene glycol 4000.

[0027] The present invention also provides a pharmaceutical formulation which is prepared by mixing the aforementioned albendazole nanocrystal lyophilized solid powder with pharmaceutically acceptable excipients.

[0028] The dosage form is a tablet, capsule, pellet, or suspension.

[0029] In this invention, the "liquid-to-bead ratio" is the ratio of the suspension to the grinding beads used in the wet media grinding process, and the unit is "v:w".

[0030] Beneficial effects:

[0031] 1. The albendazole nanocrystals prepared by this invention improve the dissolution rate of albendazole, have good stability, and can be mass-produced, providing a good intermediate for the preparation of other albendazole formulations.

[0032] 2. This invention realizes a process for preparing albendazole nanosuspension using wet media grinding. The preparation process is simple and feasible, does not use organic solvents, has low production costs, is controllable, facilitates large-scale industrial production, and is beneficial to market promotion.

[0033] 3. The albendazole nanocrystals prepared by this invention have small particle size, require less stabilizer, and have a high drug loading.

[0034] 4. The albendazole nanocrystals prepared by this invention significantly improve the solubility and bioavailability of albendazole; they can be used to prepare various albendazole formulations and have good application prospects. Attached Figure Description

[0035] Figure 1 The results of the investigation on the effect of stabilizer type on the particle size of albendazole nanocrystals;

[0036] Figure 2 The results of the investigation into the effect of grinding speed on nanocrystal size;

[0037] Figure 3 The results of the investigation into the effect of grinding time on nanocrystal size;

[0038] Figure 4 The results of the investigation on the effect of liquid droplet ratio on nanocrystal size;

[0039] Figure 5 Results of the short-term stability of albendazole nanocrystals at 4°C and room temperature;

[0040] Figure 6 Scanning electron microscope images of albendazole raw material (A) and nanocrystals (B);

[0041] Figure 7 X-ray diffraction (XRPD) curves of albendazole, a physical mixture of albendazole and stabilizer, and albendazole nanocrystalline powder;

[0042] Figure 8 The blood concentration-time curves of albendazole raw material and nanocrystals in Example 1 are shown. Detailed Implementation

[0043] The technical solution of the present invention will be described in detail below through specific embodiments, but the scope of protection of the present invention is not limited to the embodiments described.

[0044] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.

[0045] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods. Unless otherwise specified, the experimental materials used in the following embodiments are commercially available products.

[0046] Albendazole raw material was purchased from Guangxi Yiyuan Biotechnology Co., Ltd. (purity > 98%).

[0047] Tea saponin was purchased from Hunan Hanqing Biotechnology Co., Ltd. (purity ≥90%).

[0048] Betaine glycosides were purchased from Xi'an Jushengyuan Biotechnology Co., Ltd. (purity ≥90%).

[0049] Example 1: Preparation of Albendazole Nanocrystals

[0050] (1) Mix tea saponin and water thoroughly and stir with a magnetic stirrer to form stabilizer solution A;

[0051] (2) According to the mass ratio of albendazole raw material to tea saponin of 5:3, albendazole was added to the stabilizer solution A obtained in step (1), and stirred to suspend the drug in it, so as to obtain albendazole crude suspension B.

[0052] (3) The albendazole coarse suspension B obtained in step (2) was transferred to the grinding jar of a planetary ball mill. Zirconia grinding beads with a diameter of 0.25-0.3 mm were used for wet grinding. After shaking evenly, the coarse suspension B and the zirconia grinding beads were added to the grinding jar at a ratio of 1:4 (v / w). The mixture was ground at 500 rpm for 6 hours. After grinding, the grinding beads were separated to obtain an albendazole nano-suspension, i.e., albendazole nanocrystals. The concentration of albendazole in the albendazole nano-suspension was 1.0%.

[0053] The albendazole nanocrystals have an average particle size of 186.9±1.3 nm, a PDI of 0.187±0.006, and a zeta potential of -25.3±0.5 mV.

[0054] Example 2: Preparation of Albendazole Nanocrystals

[0055] (1) Mix tea saponin and water thoroughly and stir with a magnetic stirrer to form stabilizer solution A;

[0056] (2) According to the mass ratio of albendazole raw material to tea saponin stabilizer of 10:1, albendazole is added to the stabilizer solution A obtained in step (1), and stirred to suspend the drug in it, so as to obtain albendazole crude suspension B.

[0057] (3) The albendazole coarse suspension B obtained in step (2) was transferred to the grinding jar of a planetary ball mill. Zirconia grinding beads with a diameter of 0.25-0.3 mm were used for wet grinding. After shaking evenly, the coarse suspension B and the zirconia grinding beads were added to the grinding jar at a ratio of 1:4 (v / w). The mixture was ground at 500 rpm for 6 hours. After grinding, the grinding beads were separated to obtain an albendazole nano-suspension, i.e., albendazole nanocrystals. The concentration of albendazole in the albendazole nano-suspension was 1.0%.

[0058] The albendazole nanocrystals have an average particle size of 330.9±6.7 nm, a PDI of 0.236±0.010, and a zeta potential of -25.1±1.2 mV.

[0059] Example 3: Preparation of Albendazole Nanocrystals

[0060] (1) Mix tea saponin and water thoroughly and stir with a magnetic stirrer to form stabilizer solution A;

[0061] (2) According to the mass ratio of albendazole raw material to tea saponin stabilizer 1:1, albendazole is added to the stabilizer solution A obtained in step (1), and stirred to suspend the drug in it, so as to obtain albendazole crude suspension B.

[0062] (3) The albendazole coarse suspension B obtained in step (2) was transferred to the grinding jar of a planetary ball mill. Zirconia grinding beads with a diameter of 0.25-0.3 mm were used for wet grinding. After shaking evenly, the coarse suspension B and the zirconia grinding beads were added to the grinding jar at a ratio of 1:4 (v / w). The mixture was ground at 500 rpm for 6 hours. After grinding, the grinding beads were separated to obtain an albendazole nano-suspension, i.e., albendazole nanocrystals. The concentration of albendazole in the albendazole nano-suspension was 1.0%.

[0063] The albendazole nanocrystals have an average particle size of 216.0±7.5 nm, a PDI of 0.205±0.009, and a zeta potential of -22.2±0.4 mV.

[0064] Example 4: Preparation of Albendazole Nanocrystals

[0065] (1) Mix tea saponin and water thoroughly and stir with a magnetic stirrer to form stabilizer solution A;

[0066] (2) According to the mass ratio of albendazole raw material to tea saponin stabilizer 5:6, albendazole is added to the stabilizer solution A obtained in step (1), and stirred to suspend the drug in it, so as to obtain albendazole crude suspension B.

[0067] (3) The albendazole coarse suspension B obtained in step (2) was transferred to the grinding jar of a planetary ball mill. Zirconia grinding beads with a diameter of 0.25-0.3 mm were used for wet grinding. After shaking evenly, the coarse suspension B and the zirconia grinding beads were added to the grinding jar at a ratio of 1:4 (v / w). The mixture was ground at 500 rpm for 6 hours. After grinding, the grinding beads were separated to obtain albendazole nano-suspension, i.e., albendazole nanocrystals. The concentration of albendazole in the albendazole nano-suspension was 0.5%.

[0068] The albendazole nanocrystals have an average particle size of 215.2±1.2 nm, a PDI of 0.277±0.012, and a zeta potential of -21.3±3.0 mV.

[0069] Example 5: Preparation of Albendazole Nanocrystals

[0070] (1) Mix tea saponin and water thoroughly and stir with a magnetic stirrer to form stabilizer solution A;

[0071] (2) According to the mass ratio of albendazole raw material to tea saponin stabilizer 25:6, albendazole was added to the stabilizer solution A obtained in step (1), and stirred to suspend the drug in it, so as to obtain albendazole crude suspension B.

[0072] (3) The albendazole coarse suspension B obtained in step (2) was transferred to the grinding jar of a planetary ball mill. Zirconia grinding beads with a diameter of 0.25-0.3 mm were used for wet grinding. After shaking evenly, the coarse suspension B and the zirconia grinding beads were added to the grinding jar at a ratio of 1:4 (v / w). The mixture was ground at 500 rpm for 6 hours. After grinding, the grinding beads were separated to obtain albendazole nano-suspension, i.e., albendazole nanocrystals. The concentration of albendazole in the albendazole nano-suspension was 2.5%.

[0073] The albendazole nanocrystals have an average particle size of 246.2±26.4 nm, a PDI of 0.138±0.018, and a zeta potential of -24.3±0.8 mV.

[0074] Comparative Example 1

[0075] Using poloxamer 407 as a stabilizer, and with an albendazole active pharmaceutical ingredient to stabilizer mass ratio of 5:3, nanocrystals were prepared according to the parameters of Example 1. The resulting albendazole nanocrystals had an average particle size of 377.9 ± 4.8 nm, a PDI of 0.236 ± 0.031, and a zeta potential of -13.3 ± 0.4 mV. This indicates that the average particle size and PDI of the nanocrystals prepared using poloxamer 407 as a stabilizer are higher than those prepared using tea saponin as a stabilizer, and the absolute value of the zeta potential of the nanocrystals prepared using poloxamer 407 as a stabilizer is lower than that of the nanocrystals prepared using tea saponin as a stabilizer.

[0076] Example 6: Screening of Stabilizer Types

[0077] Albendazole raw material and different stabilizers were weighed at a mass ratio of 5:3. Distilled water was added to make the concentration of albendazole in the suspension 1.0%. The mixture was stirred with a magnetic stirrer to obtain a coarse suspension. The coarse suspension and 0.25-0.3 mm diameter zirconia grinding beads were transferred to the grinding jar of a planetary ball mill at a ratio of 1:4 (v / w) and ground at 500 rpm for 6 hours. After grinding, the grinding beads were separated to obtain albendazole nano suspension.

[0078] The stabilizers selected were glycyrrhizic acid (GA), tea saponin (TS), and stevia (RUB). The particle size and polydispersity index (PDI) of each nano-suspension were determined using a particle size potentiometer.

[0079] The results are as follows Figure 1 As shown, among the three stabilizers, the albendazole nanosuspension with tea saponin as the stabilizer was found to have better particle size and PDI, with the particle size order being: tea saponin (TS) < glycyrrhizic acid (GA) < stevia glycoside (RUB).

[0080] Example 7: Grinding Speed ​​Investigation

[0081] Zirconia beads with a diameter of 0.25-0.3 mm were used for wet milling. The mass ratio of albendazole raw material to stabilizer was 10:3. The concentration of albendazole in the albendazole nano suspension was 1.0%. After shaking evenly, the suspension and zirconia grinding beads were added to the grinding jar at a ratio of 1:4 (v / w) and milled at 300-600 rpm for 6 hours. The average particle size and PDI were used as evaluation indicators to investigate different milling speeds.

[0082] like Figure 2 As shown, the grinding effect was better at 500 rpm.

[0083] Example 8: Grinding Time Investigation

[0084] Zirconia beads with a diameter of 0.25-0.3 mm were used for wet milling. The mass ratio of albendazole raw material to stabilizer was 10:3. The concentration of albendazole in the albendazole nano suspension was 1.0%. After shaking evenly, the suspension and zirconia grinding beads were added to the grinding jar at a ratio of 1:4 (v / w) and milled at 500 rpm for 3-7 hours. The average particle size and PDI were used as evaluation indicators to investigate different milling times.

[0085] like Figure 3 As shown, grinding for 6 hours yielded better results.

[0086] Example 9: Investigation of Liquid Droplet Ratio

[0087] Zirconia beads with a diameter of 0.25-0.3 mm were used for wet milling. The mass ratio of albendazole raw material to stabilizer was 10:3. The concentration of albendazole in the albendazole nano suspension was 1.0%. After shaking evenly, the suspension and zirconia grinding beads were added to the grinding jar at a ratio of 1:2, 1:4, and 1:8 (v / w). The mixture was ground at 500 rpm for 6 hours. The average particle size and PDI were used as evaluation indicators to investigate different liquid-to-bead ratios.

[0088] like Figure 4 As shown, a liquid droplet ratio of 1:4 was found to produce a better grinding effect.

[0089] Example 10: Stability Study of Albendazole Nanosuspension

[0090] The albendazole nanosuspension prepared in Example 1 was placed at 4°C and room temperature for 28 days. Small amounts of the nanocrystal suspension were taken at the same time points on days 0, 7, 14, 21, and 28. The particle size, PDI, and ζ-potential were measured using a Malvern laser particle size analyzer to investigate the short-term stability over 28 days. The experimental results are shown in [Figure number missing]. Figure 5 .

[0091] in, Figure 5 A represents the particle size and PDI result of albendazole nanosuspension at 4℃; Figure 5 B represents the zeta potential of albendazole nanosuspension at 4℃; Figure 5 C represents the particle size and PDI result of albendazole nanosuspension at room temperature; Figure 5 D represents the zeta potential of albendazole nanosuspension at room temperature.

[0092] Depend on Figure 5It can be seen that the average particle size, PDI, and zeta potential of the nano-suspension on day 0 are 186.2±0.7 nm, 0.202±0.011, and -25.1±0.8 mV, respectively. At 4℃ and room temperature, the average particle size and PDI of albendazole nanocrystals remained stable at approximately 186-189 nm and 0.195-0.205, respectively, over 28 days. At 4℃, the absolute value of the zeta potential was above 23 mV, indicating good stability. At room temperature, the zeta potential decreased to some extent, but the absolute value remained above 20 mV.

[0093] Example 11: Screening of Lyophilization Protectants

[0094] Add an appropriate amount of freeze-drying protectant to the albendazole nanosuspension prepared in Example 1, mix well, transfer to a vial, pre-freeze at -80°C for 12 hours, and then freeze-dry in a freeze dryer at a vacuum of 20 Pa and -80°C for 48 hours to obtain nanocrystalline solid powder.

[0095] After screening lactose, mannitol, PEG4000, and lactose-mannitol (1:1, 1:2), lactose was found to be the most suitable freeze-drying protectant for the appearance, redispersibility, and particle size of the nanocrystalline freeze-dried powder after reconstitution. Therefore, lactose was selected as the freeze-drying protectant.

[0096] Example 12 Characterization of Albendazole Nanocrystals

[0097] (1) Scanning electron microscopy (SEM)

[0098] The morphology of albendazole raw material and the nanocrystals prepared in Example 1 were analyzed using scanning electron microscopy. The results are shown in the figure. Figure 6 .in, Figure 6 A is a scanning electron microscope image of albendazole raw material. Figure 6 B is a scanning electron microscope image of albendazole nanocrystals. The image shows that the nanocrystals are uniformly sized, near-spherical nanoparticles, with a significantly smaller particle size than the active pharmaceutical ingredient.

[0099] (2) Powder X-ray diffraction analysis (XRPD)

[0100] Albendazole raw material, a physical mixture (raw material and stabilizer), and abendazole nanocrystals prepared in Example 1 were taken separately. Powder X-ray diffraction (XRD) patterns were determined using a Cu-Kα target at a wavelength of 1.540562 nm, a tube voltage of 40 kV, a tube current of 40 mA, a scanning range of 3–40° (2θ range), a scanning wavelength of 10° / min, and a step size of 0.02°. The XRD curves are shown below. Figure 7 As shown.

[0101] As can be seen from the figure, the characteristic peaks of albendazole at 2θ (6.8°, 11.2°, 17.9°, and 24.7°) remain basically unchanged in position in the nanocrystalline powder, but the peak intensity is significantly reduced. This may be because the particle size is reduced after preparation into nanocrystals, part of the drug retains its original crystal form, while the crystallinity of another part decreases and transforms into an amorphous form, thus requiring less energy to overcome the lattice forces.

[0102] Example 13 Study on the saturated solubility of albendazole nanocrystals

[0103] Excess amounts of the active pharmaceutical ingredient (API), the physical mixture (API and stabilizer), and the nanocrystals prepared in Example 1 were added to water, pH 1.0 hydrochloric acid solution, pH 4.5 acetate buffer, and pH 6.8 phosphate buffer, respectively. The solutions were placed in a 37°C constant-temperature shaking incubator and shaken at 150 rpm for 48 hours to obtain a supersaturated solution. The solution was centrifuged at 10,000 rpm for 10 minutes, and the supernatant was collected, filtered, and analyzed by HPLC. As shown in Table 1, compared with the API and the physical mixture, the saturated solubility of the albendazole nanocrystals prepared in Example 1 of this invention was 1700–2500 times that of the API in water, pH 4.5 acetate buffer, and pH 6.8 phosphate buffer, and 3 times that of the API in pH 1.0 hydrochloric acid solution.

[0104] Table 1. Saturated solubility of active pharmaceutical ingredients, physical mixtures, and nanocrystals (37℃) (n=3)

[0105] medium Active pharmaceutical ingredient (μg / mL) Physical mixture (μg / mL) Nanocrystals (μg / mL) water 0.62±0.07 14.56±1.31 <![CDATA[1602.67±58.05 *** ]]> pH 1.0 hydrochloric acid solution 1165.06±2.14 1216.60±7.53 <![CDATA[3410.57±27.15 *** ]]> pH 4.5 acetate buffer 0.88±0.02 11.60±3.18 <![CDATA[1584.68±40.63 *** ]]> pH 6.8 phosphate buffer 0.62±0.03 7.89±1.64 <![CDATA[1604.32±24.03 *** ]]>

[0106] *** p<0.001, comparison between the nanocrystalline group and the active pharmaceutical ingredient group

[0107] Example 14: In vitro intestinal absorption study of albendazole nanocrystals

[0108] Six healthy male SD rats, 5 weeks old, weighing 180-200g, were provided by Jiangsu Qinglongshan Biotechnology Co., Ltd.

[0109] Before the experiment, rats were fasted for 12 hours but allowed free access to water. Male SD rats were randomly divided into two groups: the raw material group and the nanocrystal group. After anesthetizing the rats, the abdominal cavity was opened along the midline, and approximately 10 cm segments of the duodenum, jejunum, ileum, and colon were separated. Each segment was inverted so that the mucosa faced outward and the serosa faced inward. One end of each intestinal segment was ligated to form a sac, which was then placed in KR buffer at 37°C for 10 min to equilibrate. The other end was fixed to the sampling port, and 1.5 mL of blank KR buffer at 37°C was added. The intestinal sac was placed vertically in a test tube containing 40 mL of KR buffer (albendazole concentration 0.2 mg / mL), and a mixture of 95% O2 and 5% CO2 was continuously bubbled into the test tube to maintain the viability of the intestinal sac. 0.2 mL samples were taken from the intestinal sac at 30, 60, 90, and 120 min, and the same volume of blank KR buffer was added simultaneously. After the experiment, the length L and inner diameter R of each intestinal segment were measured. The drug concentration in the intestinal sac sample solution was determined by HPLC at each time point, and the apparent permeability coefficient P of each intestinal segment was calculated. app The experimental results are shown in Table 2. The apparent permeability coefficient P of the nanocrystal group in each intestinal segment is shown in Table 2. app Significantly improved compared to the active pharmaceutical ingredient group.

[0110] Table 2 Comparison of intestinal absorption results between nanocrystals and active pharmaceutical ingredients (n=3)

[0111]

[0112] ** p<0.01, comparison between nanocrystalline group and drug substance group

[0113] Example 15: Pharmacokinetic Study of Albendazole Nanosuspension

[0114] 1. Animal grouping and administration

[0115] One hundred healthy male ICR mice, 5 weeks old, weighing 20-30g, were provided by Jiangsu Qinglongshan Biotechnology Co., Ltd.

[0116] Protocol: The patients were randomly divided into two groups. Patients fasted for 12 hours prior to administration, but had free access to water. Albendazole and the albendazole nanosuspension prepared in Example 1 were administered by gavage, respectively, at a dose of 50 mg / kg.

[0117] 2. Sample collection:

[0118] Blood samples were collected from the fundus venous plexus at 0.5, 1, 2, 3, 4, 6, 8, 10, 12, and 24 hours after drug administration. The samples were placed in pre-heparinized EP tubes, centrifuged at 12,000 rpm for 10 min at 4°C, and the plasma was separated and placed in labeled EP tubes and stored at -80°C.

[0119] 3. Plasma Sample Processing Methods

[0120] Take 100 μL of plasma sample, add 30 μL of 1 μg / mL mebendazole internal standard solution, vortex mix, then add 600 μL of methanol, vortex thoroughly for 3 min to precipitate protein, centrifuge at 10000 r / min for 10 min, collect the supernatant, dry under nitrogen in a water bath at 45℃, reconstitute the residue with 100 μL of initial mobile phase acetonitrile-0.1% acetic acid water (20:80, v / v), vortex thoroughly, centrifuge at 10000 r / min for 10 min, collect the supernatant, perform HPLC quantitative analysis, determine plasma drug concentration, and plot blood drug concentration-time curve.

[0121] 4. Data Analysis

[0122] The pharmacokinetic data of albendazole metabolite albendazole sulfoxide were processed using Phoenix WinNonlin 8.1.0 pharmacokinetic software, and the pharmacokinetic parameters AUC and T were calculated. max C max wait.

[0123] 5. Pharmacokinetic curve after administration

[0124] After plasma processing, the pharmacokinetic curves of albendazole metabolite albendazole sulfoxide (ABZSO) at various time points were obtained (e.g., ...). Figure 8 (As shown). Peak plasma concentration C of ABZSO in the nanocrystalline group. max The AUC was significantly higher than that of the active pharmaceutical ingredient group, indicating a significant increase in the oral bioavailability of the drug and an increase in drug absorption.

[0125] 6. Pharmacokinetic parameters

[0126] After fitting with a non-compartmental model, the main pharmacokinetic parameters are shown in Table 3. The results indicate that, compared to the active pharmaceutical ingredient (API) group, the plasma drug concentration in the nanocrystal group was significantly increased, and the C0.05 of ABZSO in the nanocrystal group was significantly higher. max It is 5.0 times that of the active pharmaceutical ingredient; ABZSO's AUC 0→t The concentration was 4.7 times that of the active pharmaceutical ingredient, indicating that the nanocrystals prepared with tea saponin as a stabilizer effectively increased the oral bioavailability of the drug in vivo.

[0127] Table 3. Comparison of pharmacokinetic results between nanocrystals and active pharmaceutical ingredient (n=5)

[0128] parameter unit raw materials Nanocrystals <![CDATA[T max ]]> h 2 1 <![CDATA[C max ]]> mg / L 4.91 24.46 <![CDATA[AUC 0→t ]]> h*mg / L 34.92 162.55 <![CDATA[AUC 0→∞ ]]> h*mg / L 35.46 163.88

[0129] As described above, although the invention has been shown and described with reference to specific preferred embodiments, it should not be construed as limiting the invention itself. Various changes in form and detail may be made without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. A highly bioavailable albendazole nanocrystal, characterized in that, The nanocrystals are prepared by mixing albendazole, a naturally derived glycoside stabilizer, and water to form a suspension, and then grinding the suspension using a wet medium milling process; the average particle size is 100–500 nm. The naturally derived glycoside stabilizer is selected from tea saponins; The mass ratio of albendazole to the naturally derived glycoside stabilizer is 10:1 to 10:12; the concentration of albendazole in the suspension is 0.2% to 3.0%. The nanocrystals were prepared by the following method: (1) Thoroughly mix the natural glycoside stabilizer with water to form stabilizer solution A; (2) Add albendazole to the stabilizer solution A obtained in step (1), stir to suspend the drug therein, and obtain albendazole crude suspension B; (3) The albendazole crude suspension B obtained in step (2) is subjected to wet medium milling to obtain albendazole nano suspension, namely the albendazole nano crystals; The process parameters for wet media grinding are as follows: grinding time 2-8 hours, grinding speed controlled at 300-700 rpm, and liquid droplet ratio 1:(2-8).

2. The highly bioavailable albendazole nanocrystals according to claim 1, characterized in that, The albendazole nanocrystals have an average particle size of 100–300 nm. ζ The potential range is -30 to -20 mV.

3. The highly bioavailable albendazole nanocrystals according to claim 1, characterized in that, The mass ratio of albendazole to the natural glycoside stabilizer is 5:1 to 5:4; the concentration of albendazole in the suspension is 0.5% to 2.5%.

4. A method for preparing highly bioavailable albendazole nanocrystals according to any one of claims 1 to 3, characterized in that, Includes the following steps: (1) Thoroughly mix the natural glycoside stabilizer with water to form stabilizer solution A; (2) Add albendazole to the stabilizer solution A obtained in step (1), stir to suspend the drug therein, and obtain albendazole crude suspension B; (3) The albendazole crude suspension B obtained in step (2) is subjected to wet medium milling to obtain albendazole nano suspension, namely the albendazole nano crystals; The process parameters for the wet media grinding are as follows: grinding time 2-8 h, grinding speed controlled at 300-700 rpm, and liquid droplet ratio 1:(2-8).

5. An albendazole nanocrystalline lyophilized solid powder, characterized in that, It is prepared by freeze-drying the albendazole nanocrystals as described in any one of claims 1 to 3.

6. A pharmaceutical preparation, characterized in that, It is prepared by mixing the albendazole nanocrystal lyophilized solid powder of claim 5 with pharmaceutically acceptable excipients.

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