Posaconazole dry powder inhalant and preparation method thereof

Abstract

The invention provides a posaconazole dry powder inhalant and a preparation method thereof, and relates to the technical field of medicine preparation. The posaconazole dry powder inhalant provided by the invention comprises posaconazole, a space stabilizer and a surfactant, has excellent aerodynamic performance, good redispersibility, higher solubility and dissolution rate and excellent antifungal activity, and in-vivo pharmacokinetic evaluation shows that the lung concentration is higher and the whole-body exposure is lower. The posaconazole dry powder inhalant is prepared by adopting a wet medium grinding and spray drying technology, and a nanocrystal technology is combined with pulmonary administration, so that the obtained posaconazole dry powder inhalant has excellent aerodynamic performance, good redispersibility, higher solubility and dissolution rate, and can be used for preparing a medicine for preventing and treating diseases. And a strategy with a wide prospect is provided for targeted delivery of a drug with poor solubility to an infected part deep in the lung.

Description

Technical Field

[0001] The present application relates to the technical field of drug preparation, and in particular to a posaconazole dry powder inhaler and a preparation method thereof. Background Art

[0002] Pulmonary fungal infections have a high mortality rate for immunocompromised patients. Common pathogenic fungi include Aspergillus, Cryptococcus, Myxomyces, Fusarium, Coccidioides, and Histoplasma. Aspergillus infection is particularly prominent, with a mortality rate of up to 30%-80%. Invasive pulmonary aspergillosis (IPA) is mainly treated with triazole drugs such as itraconazole, voriconazole, and posaconazole (PSZ). PSZ is a second-generation triazole drug with broad-spectrum antifungal activity. It is approved for the prevention of invasive Aspergillus infections, the treatment of oropharyngeal candidiasis, and as a salvage therapy for rare pathogens. It inhibits the formation of ergosterol, leading to fungal cell death. The advantages of PSZ include low cross-resistance potential, few drug interactions, and activity against pathogenic yeasts and molds, all of which help improve clinical efficacy and reduce adverse reactions. However, systemic administration of PSZ results in low lung concentrations and large systemic side effects, which leads to increased doses and increased risks of hepatotoxicity and hypokalemia.

[0003] Pulmonary drug delivery systems such as dry powder inhalers (DPIs) can deliver drugs directly to the site of infection, reducing dosage and systemic side effects, thus providing an effective alternative for IPA treatment. Compared with pressurized metered dose inhalers and nebulizers, dry powder inhalers have advantages such as better stability, portability, ease of use, and efficient deposition. The solid form of PSZ makes it suitable for the preparation of dry powder inhalers, which can provide higher drug loading and greater stability. Solubility, dissolution, and aerodynamic properties are crucial for high deposition and rapid dissolution in the lower respiratory tract. The development of PSZ dry powder inhalers in existing technologies faces challenges such as low water solubility (<1μg / mL) and poor aerodynamic properties. Summary of the invention

[0004] The purpose of the present application is to provide a posaconazole dry powder inhaler and a preparation method thereof, aiming to solve the problems of low water solubility and poor aerodynamic performance of the existing posaconazole dry powder inhaler.

[0005] To achieve the above objectives, the present application provides a posaconazole dry powder inhaler, comprising: posaconazole, a steric stabilizer and a surfactant, wherein the mass percentage of the posaconazole is greater than or equal to 45% and less than 50%, the mass percentage of the steric stabilizer is greater than or equal to 45% and less than 50%, and the mass percentage of the surfactant is greater than 0% and less than 10%.

[0006] In some embodiments, the redispersibility index of the posaconazole dry powder inhaler is 1.0-1.6; the fine particle fraction of the posaconazole dry powder inhaler is 60%-80%; and the mass median of the aerodynamic diameter of the posaconazole dry powder inhaler is 2.2-3.4 μm.

[0007] In some embodiments, the zeta potential of the suspension of posaconazole dry powder inhaler is between -28.40 mV and -33.60 mV; The particle size of the suspension of the posaconazole dry powder inhaler is less than 500 nm.

[0008] In some embodiments, at least one of the following features is met: A. The posaconazole dry powder inhaler 50 2.50~4.65μm; B. The posaconazole dry powder inhaler ae 1.00~2.20μm; C. The particle size distribution Span of the posaconazole dry powder inhaler is 2.20-2.50; D. The bulk density of the posaconazole dry powder inhaler is 0.17-0.23 g / mL; E. The tap density of the posaconazole dry powder inhaler is 0.29-0.36 g / mL.

[0009] In some embodiments, the steric stabilizer is selected from HPMC, HPC or PVPK30; and the surfactant is selected from SDS.

[0010] Among them, HPMC is hydroxypropyl methylcellulose, HPC is hydroxypropyl cellulose, PVPK30 is polyvinyl pyrrolidone, and SDS is sodium dodecyl sulfate.

[0011] In some embodiments, the steric stabilizer is selected from HPMC.

[0012] The present application also provides a method for preparing the posaconazole dry powder inhaler as described above, comprising: mixing posaconazole, a steric stabilizer, a surfactant and water to obtain a mixed solution; performing wet medium grinding on the mixed solution to obtain a posaconazole suspension; The posaconazole suspension is spray-dried to obtain a posaconazole dry powder inhaler.

[0013] In some embodiments, at least one of the following features is met: A. The rotation speed of the wet medium grinding is 2000-3000 rpm; B. The grinding time of the wet medium grinding is 1 to 5 hours; C. The particle size of the grinding beads in the wet medium grinding is 0.1 mm to 0.6 mm; D. The grinding medium of the wet media grinding is zirconium oxide; E. The filling rate of the grinding medium in the wet medium grinding is 80% to 90%; F. The ratio of the sum of the masses of the posaconazole, the steric stabilizer and the surfactant (in g) to the volume of the water (in ml) is 1.0% to 1.1%.

[0014] In some embodiments, the pre-grinding is performed at 2000 rpm for 45 minutes using 0.6 mm grinding beads, and then at 3000 rpm for 40 minutes using 0.1 mm grinding beads.

[0015] In some embodiments, the spray drying gas atomization flow rate is 40-60 mmHg, the pump feed rate is 6%-10%, the air intake volume is 100%, the inlet temperature is 110°C-150°C, and the outlet temperature is 84°C-94°C.

[0016] Compared with the prior art, the beneficial effects of this application include: The posaconazole dry powder inhaler provided in the present application includes posaconazole, a steric stabilizer and a surfactant, has excellent aerodynamic properties, good redispersibility, higher solubility and dissolution rate, has outstanding antifungal activity, and in vivo pharmacokinetic evaluation shows higher lung concentration and lower systemic exposure.

[0017] The preparation method of posaconazole dry powder inhalation provided in the present application adopts wet media grinding and spray drying technology, and combines nanocrystal technology with pulmonary administration. The obtained posaconazole dry powder inhalation has excellent aerodynamic properties, good redispersibility, higher solubility and dissolution rate, and provides a promising strategy for targeted delivery of poorly soluble drugs to infection sites deep in the lungs. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope of the present application.

[0019] Figure 1 The particle size distribution diagram of NS0-NS4 prepared in Example 1; Figure 2 The zeta potential diagram of NS0-NS4 prepared in Example 1; Figure 3 The particle size distribution diagram of the PSZ NS prepared in Example 1 with the addition of a stabilizer or an active agent alone; Figure 4 The zeta potential diagram of the PSZ NS prepared in Example 1 with the addition of a stabilizer or an active agent alone; Figure 5 SEM images of NS0-NS4 prepared in Example 1; Figure 6 DSC curves of NS0-NS4 prepared in Example 1; Figure 7 is the solubility of NS0-NS4 prepared in Example 1; Figure 8 The dissolution curves of NS0-NS4 prepared in Example 1; Fig. 9 is the particle size of PSZ NP DPIs prepared in Example 6; Fig.10 is the density of PSZ NP DPIs prepared in Example 6; Fig.11 is the SEM image of PSZ NP DPIs prepared in Example 6; Fig.12 is the particle size distribution curve of PSZ NP DPIs prepared in Example 6; Fig.13 DSC curve of PSZ NP DPIs prepared in Example 6; Fig.14 is the PXRD pattern of PSZ NP DPIs prepared in Example 6; Fig.15 is the infrared spectrum of PSZ NP DPIs prepared in Example 6; Fig.16 A is a schematic diagram of the structure of NGI combined with a breathing simulator, and B is the inspiratory curve of a healthy adult and a patient; Fig.17 This is the in vitro lung deposition profile of the PSZ NP DPIs prepared in Example 6; Fig.18 The particle size (A) and zeta potential (B) of the PSZ NP DPIs after redispersion prepared in Example 6; Fig.19 is the redispersibility index of the PSZ NP DPIs prepared in Example 6; Fig. 20 is the dissolution curve of PSZ NP DPIs prepared in Example 6; Fig.21The FPF (A) and aerodynamic particle size distribution (B) of NP1DPIs under different inhalation curves; Fig. 22 For PSZ, NP1DPIs, physical mixtures and Noxafil ® MIC value of suspension; Fig.23 Noxafil was orally administered to SD rats. ® Pulmonary (A) and plasma (B) drug-time curves after suspension and pulmonary administration of NP1DPIs; Fig.24 The stability of aerodynamic particle size distribution of NP1DPIs stored in high temperature environment for 10 days; Fig.25 Stability of aerodynamic particle size distribution of NP1DPIs stored in a high humidity environment for 10 days. DETAILED DESCRIPTION

[0020] As used herein: "Prepared from" is synonymous with "comprising." As used herein, the terms "comprising," "including," "having," "containing," or any other variation thereof, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises the listed elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

[0021] The embodiments of the present application will be described in detail below in conjunction with specific examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present application and should not be considered as limiting the scope of the present application. If specific conditions are not specified in the examples, they are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0022] Example 1 Preparation of PSZ NS Tables 1 and 2 list the formulations of different posaconazole suspensions (PSZ NS), where Soluplus is a polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer. PSZ NS was prepared by wet media milling (MINIFER, Netzsch, Germany) by dispersing stabilizers and / or SDS and 1 g of PSZ powder in 200 mL of aqueous solution using a magnetic stirrer; subsequently, the suspension was pre-milled with 0.6 mm grinding beads at 2000 rpm for 45 min and then with 0.1 mm grinding beads at 3000 rpm for 40 min. Yttrium-stabilized zirconia was used as the milling media with a filling rate of 85%.

[0023] Table 1 PSZ NS formulations composed of steric stabilizers and SDS

[0024] Table 2 PSZ NS formulations with single stabilizer or SDS

[0025] Example 2 Particle size distribution and zeta potential of PSZ NS The particle size, polydispersity index (PDI) and zeta potential of the PSZ NS prepared in Example 1 were measured by dynamic light scattering (DLS) using a Zetasizer Nano ZS instrument (Malvern Ltd., UK). The sample was sonicated at 80 W for 10 s and then diluted with distilled water, and 1 mL of the suspension was added to a cuvette at 25 °C for measurement.

[0026] The particle size distribution and zeta potential of NS0~NS4 in Table 1 are as follows: Figure 1 and Figure 2 As shown. Particle size <500nm and PDI <0.25 are considered as acceptable critical quality attributes (CQAs) for NS. In addition, the absolute zeta potential value of NS must be greater than 20mV. After wet media grinding of PSZ (NS0) without the addition of any stabilizer, its particle size, PDI and zeta potential were 4.97μm, 0.46 and -11.77mV, respectively. This indicates that stabilizers are needed to reduce aggregation between drug particles and improve the stability of NS.

[0027] The particle size distribution and zeta potential of HPMC, HPC, Soluplus, PVPK30 and SDS in Table 2 are shown in Table 2. Figure 3 and Figure 4 As shown in the figure, the addition of steric stabilizer polymers (HPMC, HPC, Soluplus, and PVPK30) alone can effectively reduce the particle size compared with NS0, with the particle size ranging from 341.00 to 461.20 nm. The main reason for the particle size reduction is that the polymers and anionic surfactants are adsorbed on the surface of the dispersed nanoparticles, providing steric hindrance or electrostatic repulsion, thereby preventing the nanoparticles from agglomerating. However, the absolute zeta potential of PSZ NS using a single steric stabilizer is lower than 20 mV, which does not meet the stability requirements of NS, indicating that the steric hindrance of a single stabilizer is not sufficient to stabilize the drug particles. Although the absolute zeta potential of PSZ NS using only SDS is greater than 20 mV, its average particle size exceeds 500 nm and fails to meet the acceptable stability requirements.

[0028] In order to achieve the desired particle size, PDI and zeta potential, stabilizers and surfactants are used simultaneously. Figure 2 As shown in the figure, the zeta potential of the nanosuspensions (NS1~NS4) ranged from -28.40mV to 33.60mV, ensuring stability. The combination of steric stabilizer and SDS was significant. This combination promoted the adsorption of steric stabilizer on particles during grinding, thereby preventing nanoparticle drug aggregation. In addition, it can keep the nanosuspension in a relatively stable state by providing additional electrostatic repulsion.

[0029] Example 3 Morphology of PSZ NS The crystalline morphology of PSZNS prepared in Example 1 was observed using an HT7800TEM (Hitachi, Tokyo, Japan) with an accelerating voltage of 120 kV. Freshly prepared PSZNS were dropped on a copper grid coated with a film and incubated at room temperature for 5 minutes. The sections were negatively stained with 3% uranyl acetate and lead citrate and then air-dried for observation.

[0030] Transmission electron microscopy images of NS0~NS4 are shown in Figure 5 All PSZ nanocrystals showed a smooth rod-like morphology. NS0 without stabilizer had a larger particle size and more serious aggregation. In contrast, NS1~NS4 with stabilizer were evenly dispersed with a particle size of about 200nm. Therefore, the combination of steric stabilizer and SDS successfully prepared stable PSZ NS.

[0031] Example 4 Crystallinity of PSZ NS The PSZ NS prepared in Example 1 was centrifuged at 12,000 rpm for 20 minutes and then dried in a nitrogen environment, and then about 3-10 mg of PSZ nanoparticles were weighed and sealed in a standard aluminum crucible. The crystallinity of each sample was measured by differential scanning calorimetry (DSC) scanning using DSC-60 (Shimadzu, Japan) in the temperature range of 50-200°C at a heating rate of 10°C / min.

[0032] The DSC curves of PSZ and PSZ nanoparticles are shown in Figure 6 PSZ was crystalline with an internal thermal peak at about 168.5°C. The characteristic peaks of PSZ were observed in the thermal images of all PSZ nanoparticles, indicating that the drug nanoparticles remained crystalline after the wet media milling process, but the melting point peaks of NS1~NS4 decreased and became broader.

[0033] Example 5 Solubility and dissolution of PSZ NS 5.1 Solubility Excess PSZ (20 mg) and PSZ NS were added to phosphate buffer (PBS, pH 7.4) and supersaturated by sonication for 30 min. The samples were shaken in a water bath at 37°C ± 0.5°C for 4 h and then centrifuged at 12,000 rpm for 10 min. The supernatant was filtered through a 0.22 μm filter membrane and analyzed by HPLC. Each measurement was repeated three times.

[0034] like Figure 7 As shown in the figure, the solubility of PSZ in PBS (pH 7.4) is 5.46 µg / mL. The solubility of NS1~NS4 was significantly improved (p<0.01), ranging from 3.5 to 6.64 times that of PSZ. This increase in saturation solubility can be attributed to nanosizing, as described by the Ostwald-Freundlich equation. Reducing the drug particle size to the nanometer range can increase solubility, indicating that the dissolution properties are enhanced. It is worth noting that although NS3 has the smallest particle size ( Figure 1 ), but its solubility is the lowest among NS1~NS4. The main reason for the decrease in solubility is that NS3 is unstable in the dissolution medium.

[0035] 5.2 Dissolution The dissolution tests of PSZ and PSZNS were performed using the USP paddle method and a ZRS-8G dissolution tester (TianDa TianFa Technology Co., Ltd., China). Samples equivalent to 10 mg of PSZ were added to 200 mL of PBS containing 1% Tween 80, maintained at 37 ± 0.5 °C, and stirred at 75 rpm / min. At predetermined times (1, 3, 5, 10, 20, 30, 45, and 60 min), 1 mL of sample was drawn and passed through a 0.22 µm nylon disk filter. After each sampling, 1 mL of fresh dissolution medium was immediately added. The PSZ content in the filtrate was analyzed by HPLC. All samples were run in triplicate.

[0036] The dissolution curves of PSZ and NS1~NS4 are shown in Figure 8 The dissolution performance of PSZ was poor, with only about 28% of the drug released within 60 minutes, which was attributed to its low water solubility (<1µg / mL) and high hydrophobicity. In contrast, the dissolution rates of NS1~NS4 were significantly increased compared with PSZ (p<0.01). The improvement in dissolution may be due to the increased solubility ( Figure 7 ) and the combined effect of increased specific surface area due to reduced particle size, as described by the Noyce-Whitney equation.

[0037] Example 6 Preparation of PSZ NP DPIs by spray drying The PSZ NS (NS1-NS4) prepared in Example 1 were further spray dried using a small spray dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) with a nozzle diameter of 0.7 mm to obtain posaconazole nanocrystal agglomerated particles as dry powder inhalers (PSZ NP DPIs, NP1-NP4). The spray drying parameters were set as follows: gas atomization flow rate 50 mmHg, pump feed rate 8%, air intake 100%, inlet temperature 140°C, and outlet temperature 84-94°C. During the entire spraying process, the suspension was stirred using a magnetic stirrer. The collected powder was then stored in a glass bottle and stored in a desiccator at room temperature until use.

[0038] Example 7 Particle size and density of PSZ NP DPIs 7.1 Particle size The particle size of PSZ NP DPIs was determined by a laser diffraction particle size analyzer, and the instrument used was Sympatec HELOS&INHALER™ (SympatecGmbH, Germany). The single-dose inhaler Turbospin integrated with INHALER™ was used for dispersion. The airflow rate was set to 60L / min. The measurement took the optical concentration (Copt) of 0.5% as the trigger point and the optical concentration (Copt) below 0.5% as the endpoint. In addition, the R4 lens, which can measure the particle size range of 0.5 / 1.8-350μm, was selected to measure the particle size of the sample. Each sample was measured in triplicate. The particle size was calculated by the Fraunhofer theory in WINDOX5.0 software (Sympatec, Switzerland). The volume particle size data (d 10 d 50 and d 90 ) correspond to the equivalent spherical volume diameters of 10%, 50% and 90% cumulative volumes, respectively. The span is calculated based on the particle size distribution results.

[0039] The particle size results of NP1~NP4DPIs are shown in Fig. 9 and Table 3. d of NP1~NP4 DPIs 50 and d ae The values ​​are 2.56~4.60μm and 1.06~2.12μm respectively, both of which can be used for inhalation. 50 Value and d ae The value is the smallest, it is easier to flow and disperse in the airflow, and can be deposited in the deep areas of the lungs.

[0040] Table 3 Particle size distribution of NP1~NP4DPIs

[0041] 7.2 Bulk density and tap density The bulk density and tap density of PSZ NP DPIs were determined by manual tapping. An accurately weighed amount of powder (m) was loaded into a measuring cylinder and the bulk volume (Vb) was recorded. Subsequently, the measuring cylinder was vibrated until the powder volume did not change and the volume was then recorded as the tapped volume (Vt). The bulk density (ρb) and tapped density (ρt) were calculated as the ratio of the powder mass (m) to Vb and Vt, respectively. Each measurement was performed three times.

[0042] In addition, the aerodynamic diameter (dae) is related to the particle diameter and density as shown in formula (1); (1); Where ρ refers to bulk density, ρ0 represents unit density (equal to lg / cm 3 ), χ represents the dynamic shape factor, and d represents the particle diameter.

[0043] The bulk density (ρb) and tap density (ρt) of NP1~NP4DPIs are as follows: Fig.10 As shown. The ρb and ρt values ​​of all PSZ NP DPIs were relatively low, ranging from 0.17 to 0.23 g / mL and 0.29 to 0.36 g / mL, respectively. NP1DPIs had the lowest ρb of 0.17 ± 0.001 g / mL, which is suitable for pulmonary administration. When the particle size is similar, lower ρb can improve the aerodynamic performance of DPIs. In summary, the lower dae and density of NP1DPIs indicate that they have better aerodynamic performance.

[0044] Example 8 Morphology of PSZ NP DPIs The particle morphology of PSZ and PZS NP DPI was observed by scanning electron microscopy (SigmaHD, Zeiss, Germany). The samples were placed on double-sided conductive tape, fixed on an aluminum tube, and then gold-plated in an argon atmosphere with a thickness of 5 nm and a voltage of 5.00 kV.

[0045] SEM images of PSZ and NP1~NP4 DPIs are shown in Fig.11 As shown in Figure 2, PSZ drug particles are irregular flakes and form large agglomerates with an average particle size of about 25 μm ( Fig.12 ), particles that are too large are not suitable for inhalation. Spray drying is generally a favorable solidification technology for producing approximately spherical, single micro-sized nanocrystal agglomerates for pulmonary delivery, which are expected to exhibit better flow properties. Micrographs of NP1~NP4DPIs show that the particle size is relatively uniform, and most particles exhibit a characteristic hollow and concave morphology.

[0046] The formation of hollows and the collapse of the initially spherical particles can be attributed to the higher drying temperature (140 °C) and the increased pressure generated by the removal of water from the interior of the particles through the solid shell. In addition, the surfaces of NP1~NP4 DPIs showed that PSZ nanoparticles were embedded in the stabilizer matrix (HPMC-SDS, HPC-SDS, Soluplus-SDS, and PVPK30-SDS), which may reduce drug loss and accelerate drug release.

[0047] Example 9 Crystallinity of PSZ NP DPIs 9.1 DSC Scan About 3-10 mg of PSZ NP DPIs were weighed and subjected to DSC analysis to determine the changes in PSZ crystals and structure after wet media grinding and spray drying. The experimental procedures were as described in Example 4.

[0048] Depending on the melting point (Tm) of the drug, the DSC thermogram can provide information on whether the drug is crystalline or amorphous. Fig.13 As shown in Figure 3, the thermal map of PSZ exhibits a sharp internal heat peak at 168.5°C, indicating its crystalline nature. The characteristic melting point peak of PSZ was also observed in NP1~NP4DPIs, indicating that the crystalline nature of the drug has not changed. However, the melting point peaks of NP1~NP4DPIs decreased and became broader, indicating that their crystallinity decreased. This may be attributed to the reduction in particle size described by the Gibbs-Thomson equation and the defects and cracks induced in the crystal structure during the wet media grinding process. In addition, wet media grinding may also cause disorder in the drug lattice, thereby reducing the melting point.

[0049] 9.2 Powder X-ray Diffraction (PXRD) PXRD scans of the samples were performed in the range of 5° to 50° using a Bruker D8 advanced diffractometer (Bruker, Karlsruhe, Germany) equipped with CuKa radiation at a voltage of 40 kV, a current of 30 mA, and a speed of 10° / min.

[0050] The PXRD patterns of PSZ and NP1~NP4DPIs are shown in Fig.14As shown. PSZ showed crystallinity with several sharp diffraction peaks at 2θ values ​​of 7.7°, 9.9°, 14.4°, 15.6°, 15.9°, 17.8°, 19.4°, 20.0°, 22.3°, and 25.5°. In the diffraction patterns of NP1~NP4DPIs, the positions of all PSZ characteristic peaks remained unchanged, and no additional peaks appeared, indicating that PSZ NS did not induce any polymorphic transformation after wet media milling and spray drying. However, the diffraction peak intensity of NP1~NP4DPIs decreased compared with PSZ, which was mainly due to the reduction of drug particle size during the preparation process and the presence of stabilizers that can reduce drug aggregation. Therefore, the crystalline state of PSZ did not change during wet media milling and spray drying, but some components became amorphous.

[0051] Example 10 Fourier Transform Infrared Spectroscopy Fourier transform infrared spectroscopy (Spectrum100 Fourier transform infrared spectrometer, PerkinElmer, USA) is used to detect the compatibility or interaction between drugs and excipients. It is used to study the molecular interaction between drugs and excipients. The change in the position of the characteristic absorption band is usually related to the potential drug-excipient interaction. Take a small amount of sample and mix it with KBr. Use a tablet press to press the mixture into a disc. Place the disc on the sample holder of the Fourier transform infrared spectrometer and press it at 1000-4000cm -1 Scanning within the range with a resolution of 4cm -1 .

[0052] like Fig.15 As shown, PSZ exhibits characteristic absorption bands at 3320 cm-¹ (OH stretching vibration), 2965 cm-¹ (CH aliphatic stretching vibration), 1686 cm-¹ (C=O stretching vibration), 1512 cm-¹ (C=C stretching vibration) and 1234 cm-¹ (COC stretching vibration), which is consistent with the PSZ spectrum described in the previous literature. There is no obvious difference in the absorption band positions of PSZ and NP1~NP4DPIs, confirming that NP1~NP4DPIs did not interact during the spray drying process.

[0053] Example 11 In vitro aerodynamic properties of PSZ NP DPIs 11.1 In vitro aerodynamic performance A new generation impactor (NGI, Copley Scientific Ltd., Nottingham, UK) equipped with a suitable adapter and artificial throat was used to evaluate the aerodynamic properties of the powders. First, PSZ NP DPIs were loaded into 3# Vcaps® capsules. Then, the capsules were placed in a TurbospinTM device (PH&TS.pA, Milan, Italy) and tested for 4 s at an airflow rate of 60 L / min and a pressure drop of 4 kPa on the device. After the test, the powder deposited on the adapter, induction port and all NGI platforms was carefully cleaned with anhydrous ethanol and collected for HPLC analysis. Parameters representing aerodynamic performance, namely fine particle fraction (FPF), mass median aerodynamic diameter (MMAD) and geometric standard deviation (GSD), were calculated using CITDAS® software (version 3.10, Copley Scientific Ltd., UK).

[0054] The key aerodynamic parameters of NP1~NP4 DPIs are shown in Table 4. FPF is considered to be the percentage of recovered drug with an aerodynamic diameter below 5μm, representing the proportion of drug that is effectively deposited in the lungs and produces clinical effects. The lung deposition efficiency of NP1~NP4 DPIs is very high, with FPF ranging from 62.92% to 78.58%. Among them, NP1 DPIs have the highest FPF value, which is 1.25 times that of NP2 DPIs, indicating that their aerodynamic performance has been improved. MMAD refers to the aerodynamic diameter of particles with a mass accumulation percentage of 50%, which is calculated by the deposition of NGI. Among PSZ NP DPIs, NP1 DPIs have the smallest MMAD value (2.26μm), indicating that the particles are more likely to reach the deep lung. In addition, GSD is one standard deviation higher or lower than the MMAD value, indicating that there are differences in aerosol particle size. Most therapeutic aerosols are polydisperse, that is, GSD>1.2. All PSZ NP DPIs were polydisperse with GSD values ​​ranging from 1.72–2.04.

[0055] Table 4 Key aerodynamic parameters of NP1~NP4DPIs

[0056] 11.2 In vitro aerodynamic performance under the true inspiratory curve Because IPA patients may have reduced lung function, the aerodynamic performance of NPDPIs under a real inspiratory curve was further evaluated in vitro, using the NGI in combination with a breathing simulator to simulate the real inspiratory characteristics of healthy adults and patients. Fig.16As shown in Figure A, the NGI was connected to a BRS300i breathing simulator (Copley Scientific, Nottingham, UK). The inspiratory curves of 30 healthy adults and 10 representative patients were analyzed. Fig.16 As shown in B. Since the peak inspiratory flow of the inspiratory profile was less than 60 L / min, the airflow through the NGI was set to 60 L / min to ensure that the breathing simulator could fully reproduce the peak inspiratory flow of each profile. The side arm of the mixing inlet was then opened to provide supplementary air to the NGI at the same rate (60 L / min), so that the airflow through the adapter was zero before the start of inspiration. Computer software was used to initiate each inspiration and draw air from the supplementary air flow as the inspiration curve at the time of initiation. Each inspiration curve was performed three times. After the test, the powder deposited on the adapter, sensing port, Copley mixing inlet (for the next generation impactor), and NGI stages were collected and analyzed by HPLC.

[0057] The in vitro lung deposition distribution of NP1~NP4 DPIs is shown in Fig.17 Compared with NP2~NP4 DPIs, NP1 DPIs had a significantly lower deposition ratio in the artificial larynx and tended to deposit more in stages 2 (4.46 μm) to 6 (0.55 μm) of the NGI, indicating a greater chance of deposition in the deep lung. In conclusion, NP1 DPIs have excellent aerosol properties, which can be attributed to their smallest particle size, lower density, and less aggregation.

[0058] Example 12 Redispersibility of PSZ NP DPIs Redispersibility is one of the important quality attributes of NPs, which indicates whether the NPs can be completely redispersed into their original nanosuspension after contact with aqueous media, which is a prerequisite for PSZ release and dissolution. Redispersibility is expressed by the redispersibility index (RDI), which is calculated according to formula (2): (2); Where z-average0 represents the average particle size of the freshly prepared nanosuspension before spray drying, and z-average represents the corresponding value of the redispersed nanosuspension after spray drying. RDI close to 1 indicates that the agglomerated particles of PSZ nanocrystals can regenerate the original nanosuspension after rehydration. The particle size of these redispersed nanoparticles was measured by Nano ZS Zetasizer (Malvern Instruments Ltd., UK).

[0059] Fig.18The particle size (A) and zeta potential (B) of the redispersed NP1~NP4DPIs are shown. Except for NP3DPIs, all NP DPIs can be redispersed into particles with a size of less than 500nm after contact with aqueous media. Drugs in this particle size range can dissolve rapidly in the lungs, thereby avoiding phagocytosis by alveolar macrophages. In addition, the redispersed NP1~NP4DPIs are all in anionic states with negative zeta potential, which makes them more easily degraded in the lungs and less likely to cause infection or fibrosis due to long-term retention.

[0060] like Fig.19 As shown in the figure, NP1DPIs have excellent redispersibility with an RDI of 1.07, which indicates that they can prevent irreversible nanoparticle aggregation caused by thermal stress during the drying stage, thereby fully exerting the advantages of nanosuspensions. In contrast, NP3DPIs have poor redispersibility (RDI>4), which may be due to the inactivation of the stabilizer after spray drying, which further leads to irreversible aggregation.

[0061] Example 13 Dissolution of PSZ NP DPIs The dissolution rates of PSZ and PSZ NP DPIs were evaluated using a ZRS-8G dissolution tester, and the experimental procedures were as described in Example 5.

[0062] The dissolution curves of PSZ and NP1~NP4DPIs are shown in Fig. 20 Compared with PSZ, the dissolution rates of NP1, NP2, and NP4 DPIs were significantly increased (p < 0.01), which was mainly due to their ability to completely redisperse into the original nanosuspension and then further dissolve ( Fig. 20 According to the Noyes-Whitney equation, this is related to the small particle size and large specific surface area of ​​drug nanoparticles. In addition, the formation of PSZ nanocomposites may lead to crystal defects ( Fig.14 ), thereby increasing the surface free energy and thus improving the dissolution rate of PSZ. However, the dissolution profiles of NP3DPIs showed only a slight improvement in dissolution rate compared with PSZ, which may be due to the poor redispersibility of NP3DPIs in the dissolution medium.

[0063] The increased in vitro dissolution rate of PSZ NP DPIs may lead to an improved in vivo bioavailability of PSZ, which is particularly important for pulmonary delivery of poorly water-soluble drugs such as PSZ. Since NP1 DPIs not only have excellent redispersibility and significantly improved dissolution rate but also exhibit the best aerodynamic properties, they were considered the best formulation for further study.

[0064] Example 14 In vitro aerodynamic performance of PSZ NP1 DPIs under real inspiratory curves Fig.21 The in vitro aerodynamic performance of NP1DPIs under healthy and diseased inspiratory curves is demonstrated. Fig.21 As shown in Figure A, NP1DPIs had no significant effect on FPF values ​​under different inspiratory modes (p>0.05), indicating that actual subject breathing can produce high lung deposition similar to that of a constant flow rate inertial impactor (Table 4). More importantly, although the inspiratory flow rate and respiratory muscle function of patients were lower than those of healthy adults, this suggests that the inspiratory flow of patients was sufficient to release and decompose NP1DPIs, generating inhalable particles (1-5μm) for pulmonary delivery. Therefore, it can be concluded that patients with lung infections can achieve higher lung deposition by inhaling NP1DPIs and achieve good therapeutic effects in clinical practice.

[0065] Example 15 In vitro antifungal activity of PSZ NP1 DPIs PSZ, NP1DPIs, physical mixtures, and Noxafil ® The in vitro antifungal activity of the suspension was evaluated against the most common clinical Candida albicans and Aspergillus niger. The minimum inhibitory concentration (MIC) was determined using the microdilution method according to the National Committee for Clinical Laboratory Standards (NCCLS) M27-A and M38-P documents with slight modifications. The fungal suspension was prepared by dilution in sterile sodium chloride-peptone buffer, pH 7.0, and the concentration of the Candida albicans suspension was 5 × 10 3 CFU / mL, the concentration of Aspergillus niger suspension was 2.6×10 4 CFU / mL. PSZ, NP1DPIs, physical mixture and Noxafil ® The suspension was dissolved in dimethyl sulfoxide (DMSO) and then diluted with Sabouraud dextrose broth (SDB) to a final concentration of DMSO below 1%.

[0066] For MIC testing, 100 μL of fungal suspension and 100 μL of samples of different concentrations were added to each well of a 96-well plate. The PSZ concentration in the wells was kept constant from 0.0078 μg / mL to 8 μg / mL, and each dilution was repeated in triplicate. The same test was performed on growth controls (SDB and fungal suspension) and negative controls (SDB). The MIC was the lowest concentration at which turbidity was significantly reduced compared to the growth control (approximately 50% inhibition of Candida albicans growth and complete inhibition of Aspergillus niger growth as visually observed).

[0067] PSZ, NP1DPIs, physical mixtures, and Noxafil ® The MIC values ​​of the suspension are shown in Fig. 22. The results showed that all formulations had significant antifungal activity against Candida albicans and Aspergillus niger. The MIC values ​​of the physical mixtures did not differ from those of PSZ, indicating that the antifungal activity did not change after the addition of excipients (HPMC and SDS). Meanwhile, the MIC values ​​of NP1DPIs were consistent with those of the physical mixtures, indicating that the antifungal activity of NP1DPIs was not affected during wet media grinding and spray drying. In addition, the antifungal activity of NP1DPIs was comparable to that of the commercial formulation Noxafil. ® The suspensions were comparable, suggesting that NP1 DPIs have good promise for the treatment of pulmonary fungal infections.

[0068] Example 16 In vivo pharmacokinetics of PSZ NP1 DPIs To further explore Noxafil ® The in vivo performance of the suspension and NP1DPIs after oral and pulmonary administration, respectively, was investigated by pharmacokinetic studies.

[0069] 16.1 Experimental Procedure Male SD rats were randomly divided into two groups, 45 in each group: Noxafil ® -Oral group and NP1-inhalation group. Both groups of rats were given the same dose of 6.25mg / kg PSZ. One group of rats was orally administered 0.04mL Noxafil ® The rats in the NP1 inhalation group were anesthetized with 3% pentobarbital (w / v) intraperitoneally, and then placed on a slanted board with their front incisors fixed with nylon floss. The NP1 DPIs were placed in a DP-4R dry powder inhaler. Preliminary studies have shown that about 92% of the intended dose was successfully injected into the rat airways.

[0070] At predetermined time points after administration (5 min, 15 min, 30 min, 1 h, 3 h, 6 h, 9 h, 12 h, 24 h, and 36 h, 5 rats per group per time point), cardiac puncture was performed and blood was collected into heparinized tubes. Blood was centrifuged at 4000 rpm for 10 min at 4°C to obtain plasma. Lungs were removed and minced with surgical scissors and then transferred to a glass homogenizer filled with an appropriate amount of cold saline (3 mL / g (lung weight)). The homogenized lung samples were centrifuged at 6500 rpm for 20 min at 4°C. Plasma and lung homogenate supernatant were frozen and stored at -20°C until further analysis.

[0071] 16.2 Analysis of Plasma and Lung Samples 10 μL of internal standard (ketoconazole, 1000 ng / mL) and 10 μL of methanol were added to 100 μL of plasma or lung homogenate supernatant, followed by vortexing for 2 minutes. The resulting suspension was extracted with 1 mL of ethyl acetate and centrifuged at 12,000 rpm for 10 minutes. Then, the upper organic matter was collected and dried under a stream of nitrogen. Finally, the dry residue was resuspended with 100 μL of methanol, then vortexed for 2 minutes and centrifuged at 12,000 rpm for 10 minutes. The concentration of PSZ was determined by HPLC-MS (Accela, ThermoScientific Inc.), and the concentration values ​​at each time point were averaged to determine the concentration-time curve.

[0072] Pharmacokinetic parameters (Tmax, Cmax, T1 / 2, and AUC0→∞) were calculated using the Microsoft Excel add-on program PKSolver (version 2.0, China Pharmaceutical University). The two groups of measured values ​​that met normal distribution and variance homogeneity were tested by independent t test, otherwise the nonparametric Mann-Whitney U test was used.

[0073] Noxafil ® The lung and plasma pharmacokinetic curves of the oral group and NP1 inhalation group are shown in Fig.23 , A is the lung drug-time curve, B is the plasma drug-time curve. The pharmacokinetic parameters are shown in Table 5 and Table 6. The results show that compared with the traditional oral Noxafil ® Compared with suspensions, inhaled NP1DPIs have the advantages of wider lung distribution and lower systemic exposure.

[0074] Table 5 Pharmacokinetic parameters in the lungs of SD rats after administration

[0075] Note: **p<0.01.

[0076] Table 6 Pharmacokinetic parameters in plasma of SD rats after administration

[0077] Note: **p<0.01.

[0078] 16.3 Stability Study The stability studies of NP1DPIs were conducted in a high humidity environment at 75 ± 5% relative humidity or a high temperature environment at 60 ± 5 ° C. The in vitro aerodynamic properties of NP1DPIs were tested on days 0, 5, and 10.

[0079] Stability study data are available at Fig.24 , Fig.25and Table 7. The results showed that NP1DPIs were stable in high temperature environment (60±2°C) but unstable in high humidity environment (75±5%RH). Therefore, it is not recommended to store NP1DPI directly in a high humidity environment. After 10 days of storage at 60±2°C, the fine particle fraction (FPF) value of NP1DPI was not significantly different from that of the initial (0-day) sample (p>0.05), indicating good aerodynamic stability. However, after 10 days of storage at 75±5% RH, the FPF value of NP1DPIs decreased significantly from 72.46±3.41% to 60.82±1.61% (p<0.05) due to severe agglomeration during storage. Although instability occurs under high humidity conditions, this problem can usually be alleviated by sealing the capsules with aluminum foil.

[0080] Table 7 In vitro aerodynamic properties of NP1DPIs stored in high temperature or high humidity environment for 10 days

[0081] In this application, PSZ NPDPIs containing various stabilizers were prepared by wet media grinding and spray drying technology. These DPIs have excellent aerodynamic properties, good redispersibility, higher solubility and dissolution rate. In vitro experiments showed that NP1DPIs have excellent antifungal activity. In addition, in vivo pharmacokinetic evaluation showed that compared with Noxafil ® Compared with suspensions, NP1 DPIs have higher lung concentrations and lower systemic exposure. Nanocrystal-based DPIs combine the advantages of nanocrystal technology with pulmonary drug delivery, providing a promising strategy for targeted delivery of poorly soluble drugs to sites of infection deep in the lungs.

[0082] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present application, rather than to limit it. Although the present application has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present application.

[0083] In addition, those skilled in the art will appreciate that, although some embodiments herein include certain features included in other embodiments but not other features, the combination of features of different embodiments is meant to be within the scope of the present application and form different embodiments. For example, in the above claims, any one of the claimed embodiments may be used in any combination. The information disclosed in this background technology section is intended only to deepen the understanding of the overall background technology of the present application and should not be regarded as an admission or in any form of implication that the information constitutes prior art known to those skilled in the art.

Claims

1. A posaconazole dry powder inhaler, characterized in that: include: Posaconazole, a steric stabilizer and a surfactant, wherein the mass percentage of the posaconazole is greater than or equal to 45% and less than 50%, the mass percentage of the steric stabilizer is greater than or equal to 45% and less than 50%, and the mass percentage of the surfactant is greater than 0% and less than or equal to 10%.

2. The posaconazole dry powder inhaler according to claim 1, characterized in that: The redispersibility index of the posaconazole dry powder inhaler is 1.0-1.6; the fine particle fraction of the posaconazole dry powder inhaler is 60%-80%; and the aerodynamic diameter mass median of the posaconazole dry powder inhaler is 2.2-3.4 μm.

3. The posaconazole dry powder inhaler according to claim 2, characterized in that: The zeta potential of the suspension of the posaconazole dry powder inhaler is between -28.40 mV and -33.60 mV; The particle size of the suspension of the posaconazole dry powder inhaler is less than 500 nm.

4. The posaconazole dry powder inhaler according to claim 3, characterized in that: Satisfy at least one of the following characteristics: A. The posaconazole dry powder inhaler 50 2.50~4.65μm; B. The posaconazole dry powder inhaler ae 1.00~2.20μm; C. The particle size distribution Span of the posaconazole dry powder inhaler is 2.20-2.50; D. The bulk density of the posaconazole dry powder inhaler is 0.17-0.23 g / mL; E. The tap density of the posaconazole dry powder inhaler is 0.29-0.36 g / mL.

5. The posaconazole dry powder inhaler according to any one of claims 1 to 4, characterized in that: The steric stabilizer is selected from HPMC, HPC or PVPK30; and the surfactant is selected from SDS.

6. The posaconazole dry powder inhaler according to claim 5, characterized in that: The steric stabilizer is selected from HPMC.

7. A method for preparing the posaconazole dry powder inhaler according to any one of claims 1 to 6, characterized in that: include: mixing posaconazole, a steric stabilizer, a surfactant and water to obtain a mixed solution; performing wet medium grinding on the mixed solution to obtain a posaconazole suspension; The posaconazole suspension is spray-dried to obtain a posaconazole dry powder inhaler.

8. The preparation method according to claim 7, characterized in that: Satisfy at least one of the following characteristics: A. The rotation speed of the wet medium grinding is 2000-3000 rpm; B. The grinding time of the wet medium grinding is 1 to 5 hours; C. The particle size of the grinding beads in the wet medium grinding is 0.1 mm to 0.6 mm; D. The grinding medium of the wet media grinding is zirconium oxide; E. The filling rate of the grinding medium in the wet medium grinding is 80% to 90%; F. The ratio of the sum of the masses of the posaconazole, the steric stabilizer and the surfactant to the volume of the water is 1.0% to 1.1%.

9. The preparation method according to claim 8, characterized in that: Pre-grind with 0.6 mm grinding beads at 2000 rpm for 45 min, and then pre-grind with 0.1 mm grinding beads at 3000 rpm for 40 min.

10. The preparation method according to claim 7, characterized in that: The gas atomization flow rate of the spray drying is 40-60 mmHg, the pump feed rate is 6%-10%, the air intake is 100%, the inlet temperature is 110°C-150°C, and the outlet temperature is 84°C-94°C.

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