A paralivirine composition for inhalation, a method for preparing the same, and use thereof

Abstract

The present application relates to a kind of dry powder inhalation of peramivir, the dry powder inhalation is composed of 50-90% peramivir and pharmaceutically acceptable carrier by weight percentage, wherein, the aerodynamic diameter (MMAD) of dry powder particle that constitutes dry powder inhalation is 1 μm-5 μm, the particle size distribution of dry powder particle is D10≤2.0 μm, D50≤4.0 μm, D90≤7.5 μm.The dry powder inhalation of the present application has the advantages of high antibacterial activity, long-acting targeting, high safety and effectiveness, good stability, high bioavailability and the like.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to an inhaled peramivir composition, its preparation method, and its application. Background Technology

[0002] Influenza is an acute respiratory infectious disease caused by the influenza virus. It spreads rapidly, and the general population is susceptible. Influenza viruses belong to the Orthomyxoviridae family and are classified into four types—A, B, C, and D—based on antigenic differences in their nucleoproteins and matrix proteins. Among these, influenza A and B viruses have the most significant impact on human public health. Influenza A viruses have a wide host range, infecting humans and various animals. Their surface antigens (hemagglutinin and neuraminidase) mutate frequently and have a strong mutation capacity, often causing seasonal epidemics and even global pandemics. Currently, the main seasonally circulating subtypes of influenza A in the human population are H1N1 and H3N2. Influenza B viruses have a narrower host range, primarily infecting humans. Their antigenic mutation rate is slower, and they are divided into two main lineages: Victoria and Yamagata. They typically cause outbreaks or epidemics in localized areas, with a particularly significant impact on children.

[0003] According to global data from the World Health Organization, there are approximately 1 billion cases of seasonal influenza each year, including 3 to 5 million severe cases, and approximately 290,000 to 650,000 deaths annually from influenza-related respiratory illnesses or complications. Influenza can lead to serious consequences such as pneumonia, acute respiratory distress syndrome, myocarditis, influenza-related encephalopathy, and even multiple organ failure. A global systematic review and meta-analysis estimates that the hospitalization rate for seasonal or pandemic influenza is approximately 1.01%, the all-cause mortality rate for non-severe influenza is 0.23%, while the all-cause mortality rate for severe influenza is as high as 4.15%. In China, the influenza hospitalization rate is 73 per 100,000 people, and data from January 2025 shows an upward trend in the number of severe influenza cases, pneumonia, and influenza-related deaths. In the United States, the 2024-2025 influenza season is expected to be the most severe in over a decade, with the CDC estimating at least 43 million infections, 560,000 hospitalizations, and 24,000 deaths.

[0004] Peramivir ((1S,2S,3S,4R)-3-[(1S)-1-acetamido-2-ethyl-butyl]-4-guanidino-2-hydroxy-cyclopentyl-1-carboxylic acid) is a novel neuraminidase inhibitor (NAI) primarily used to treat influenza A and B virus infections. Peramivir works by targeting neuraminidase (NA) on the surface of the influenza virus, inhibiting viral release from host cells and thus blocking viral transmission. Structural biology studies (such as X-ray crystallography) have further elucidated its binding mode with NA: peramivir forms hydrogen bonds with conserved residues (such as E119 and R292) at the NA active site through its carboxylic acid group, while its cyclohexane structure occupies the hydrophobic pocket of NA, enhancing binding specificity and inhibitory efficiency. Compared to oseltamivir and zanamivir, peramivir maintains high activity against some NA mutant strains (such as the H275Y mutation in H1N1). Clinically, peramivir is used to treat adult influenza, gradually expanding to include children, severely / critically ill patients, and immunocompromised individuals. Furthermore, dosage regimens for children and pregnant women need further optimization (e.g., adjusting dosage according to weight), and safety data for pregnant women need to be clarified. For severely ill / immunosuppressed patients, the optimal timing and dosage of combination therapy (e.g., with hormones or immunomodulators) need to be explored, as well as the efficacy of prolonged treatment (e.g., more than 5 days), and other aspects of the influenza virus life cycle (e.g., viral entry into cells, RNA replication). Combination regimens of peramivir with RNA polymerase inhibitors (e.g., baloxavir), M2 ion channel blockers (e.g., amantadine, although resistant, have complementary mechanisms), or host factor inhibitors (e.g., targeting the PI3K / Akt pathway) are under investigation, aiming to enhance antiviral efficacy and reduce the risk of drug resistance.

[0005] (Peramivir)

[0006] Peramivir, as the only approved intravenous NAI, overcomes the limitations of oral (oseltamivir) or inhaled (zanamivir) administration in certain patients (such as infants, young children, those with swallowing difficulties, and those with impaired consciousness), especially during influenza pandemics, where intravenous administration offers greater accessibility. Multiple Phase III clinical trials (such as CAPSTONE-1 and CAPSTONE-2) have shown that peramivir's time to clinical remission (TTAS) in treating adult influenza is comparable to that of oseltamivir (approximately 21-26 hours), but it clears the virus more quickly in severely ill patients. Pediatric studies (such as a trial involving 200 children) have shown that a single 300 mg / kg infusion is as effective as oral oseltamivir and is well-tolerated.

[0007] Influenza virus resistance to NAIs primarily stems from point mutations in the NA gene (such as H275Y in H1N1 and E119V / I in H3N2). The resistance mutation spectrum of peramivir partially overlaps with that of oseltamivir, but there are differences: for example, the H275Y mutant strain of H1N1 is highly resistant to oseltamivir but remains sensitive to peramivir (IC50 increases only 2-4 times); while the E119K mutation in H3N2 may lead to decreased sensitivity to peramivir (IC50 increases more than 10 times). Data from the Global Influenza Surveillance Network (GISRS) shows that the detection rate of resistant strains after peramivir's market launch is extremely low (<1%), far lower than that of oseltamivir (in recent years, the resistance rate of H1N1 has reached 20%-30% in some seasons). With the prevalence of oseltamivir-resistant strains, peramivir, due to its unique resistance spectrum, is expected to become one of the core drugs for future influenza treatment.

[0008] However, intravenous administration of peramivir has the following drawbacks: first, it cannot be directly targeted to the lungs; second, it may increase the risk of viral infection for healthcare workers; and third, injection is inconvenient and cannot be administered independently. There is a need to develop novel antiviral agents that combine high efficacy, convenient administration, high patient accessibility, and good safety.

[0009] The deposition and distribution of dry powder inhalers (DPIs) in the lungs and respiratory system are related to their particle size distribution. Particles with a diameter > 5.0 μm mainly deposit in the pharynx and upper respiratory tract; particles with a diameter of 1.0-5.0 μm mainly deposit on the surface of the trachea, bronchi, and alveoli; particles with a diameter of 0.5-1.0 μm mainly deposit on the respiratory bronchioles and alveolar walls; particles with a diameter < 0.5 μm are virtually impossible to deposit in the respiratory tract. A higher proportion of particles with a diameter of 1.0-3.0 μm in the dry powder inhaler indicates a higher deposition rate in the alveoli and bronchioles. Currently, there are no commercially available dry powder inhalers for peramivir. Summary of the Invention

[0010] The present invention aims to provide a peramivir dry powder inhaler, wherein, by weight percentage, the dry powder inhaler is composed of 50-90% peramivir and a pharmaceutically acceptable carrier, wherein the aerodynamic diameter (MMAD) of the dry powder particles constituting the dry powder inhaler is 1μm-5μm, and the particle size distribution of the dry powder particles is D10≤2.0μm, D50≤4.0μm, and D90≤7.5μm.

[0011] In a preferred embodiment of the present invention, the pharmaceutically acceptable carrier is a combination of phospholipids and calcium chloride.

[0012] In a preferred embodiment of the present invention, the phospholipid is selected from any one or a combination of distearylphosphatidylcholine (DSPC), dipalmitoylphosphatidylcholine, dilauroylphosphatidylcholine, dimyristoylphosphatidylcholine, hydrogenated soybean phospholipid, distearylphosphatidylglycerol, and dipalmitoylphosphatidylglycerol.

[0013] In a preferred embodiment of the present invention, the molar ratio of phospholipid to calcium chloride in the pharmaceutically acceptable carrier is 1:4-4:1, preferably 1-4:1.

[0014] In a preferred embodiment of the present invention, the dry powder inhaler contains 50-90% peramivir, 1-20% distearate phosphatidylcholine, and 0.1-2% calcium chloride.

[0015] In a preferred embodiment of the present invention, the dry powder inhaler contains 70-90% peramivir, 10-20% distearate phosphatidylcholine, and 0.5-2% calcium chloride.

[0016] In a preferred embodiment of the present invention, the dry powder inhaler contains 84.82% peramivir, 14.34% distearate phosphatidylcholine, and 14.34% calcium chloride.

[0017] In a preferred embodiment of the present invention, the dry powder inhaler contains 84.13% peramivir, 84.13% distearate phosphatidylcholine, and 1.67% calcium chloride.

[0018] In a preferred embodiment of the present invention, the dry powder inhaler contains 84.83% peramivir, 84.83% distearate phosphatidylcholine, and 0.84% ​​calcium chloride.

[0019] In a preferred embodiment of the present invention, the dry powder inhaler contains 0.84% ​​peramivir, 17.9% distearate phosphatidylcholine, and 1.06% calcium chloride.

[0020] In a preferred embodiment of the present invention, the aerodynamic diameter of the dry powder particles is 1-4.5 μm, preferably 1.5-4 μm, and more preferably 2.5-3.5 μm.

[0021] In a preferred embodiment of the present invention, the particle size distribution of the dry powder particles is D10≤0.5-2μm, D50≤2.5-4μm, and D90≤5-7.5μm.

[0022] In a preferred embodiment of the present invention, the particle size distribution of the dry powder particles is D10≤1-1.5μm, D50≤3-4μm, and D90≤5.5-7μm.

[0023] In the preferred embodiment of the present invention, the effective deposition rate (FPF) of the dry powder particles is ≥50%, preferably ≥60%, and more preferably ≥70%.

[0024] In a preferred embodiment of the present invention, the emptying rate (ER) of the dry powder particles is ≥90%, preferably ≥95%.

[0025] In a preferred embodiment of the present invention, the moisture content of the dry powder particles is ≤12%, preferably ≤10%.

[0026] In a preferred embodiment of the present invention, the surface of the dry powder particles has a porous structure.

[0027] In a preferred embodiment of the present invention, the geometric deviation (GSD) of the inhaled particles is 1-3, preferably 1.5-2.7.

[0028] In a preferred embodiment of the present invention, the surface of the dry powder particles has a porous structure.

[0029] In a preferred embodiment of the present invention, the dry powder inhaler delivers a pulmonary dose of at least about 10 mg in a single inhalation via pulmonary inhalation.

[0030] In a preferred embodiment of the present invention, the preparation of the peramivir dry powder inhaler includes the following steps:

[0031] (1) Add the required amount of distearate phosphatidylcholine and calcium chloride to water at 70-80℃, and shear at high speed (3000rpm-8000rpm)* (1min-5min) to obtain an aqueous phase;

[0032] (2) Under stirring conditions (3000rpm-8000rpm), the required amount of perfluorooctane (PFOB) was added to the aqueous phase and subjected to high-speed shearing (3000rpm-8000rpm)* (1min-5min) to obtain the primary emulsion;

[0033] (3) Add the required amount of peramivir to the primary emulsion, perform high-speed shearing (3000rpm-8000rpm)*(1min-5min), high-pressure homogenization (400bar-800bar)*(1min-5min), and spray dry the emulsion to obtain the final product.

[0034] In the preferred embodiment of the present invention, the high-speed shearing conditions in steps (1), (2), and (3) are (3000rpm-8000rpm)*(1min-5min).

[0035] In the preferred embodiment of the present invention, in step (2), the mass ratio of perfluorobromooctane to distearate phosphatidylcholine is 5-55:1, preferably 8-50:1, and more preferably 10-20:1.

[0036] In a preferred embodiment of the present invention, the solid content in the emulsion is 1%-4% by mass-volume percentage, preferably 1.5%-3.5%, and more preferably 2-3%.

[0037] In a preferred embodiment of the present invention, the inlet temperature of the spray dryer is 100-180°C, preferably 120-165°C.

[0038] In a preferred embodiment of the present invention, the emulsion temperature is maintained at 4-30°C during spray drying.

[0039] Another object of the present invention is to provide a method for preparing a peramivir dry powder inhaler, wherein, by weight percentage, the dry powder inhaler consists of 50-90% peramivir and a pharmaceutically acceptable carrier, wherein the aerodynamic diameter (MMAD) of the dry powder particles constituting the dry powder inhaler is 1 μm-5 μm, and the particle size distribution of the dry powder particles is D10≤2.0 μm, D50≤4.0 μm, and D90≤7.5 μm. The preparation of the peramivir dry powder inhaler includes the following steps:

[0040] (1) Add the required amount of distearate phosphatidylcholine and calcium chloride to water at 70-80℃, and shear at high speed (3000rpm-8000rpm)* (1min-5min) to obtain an aqueous phase;

[0041] (2) Under stirring conditions (3000rpm-8000rpm), the required amount of perfluorooctane (PFOB) was added to the aqueous phase and subjected to high-speed shearing (3000rpm-8000rpm)* (1min-5min) to obtain the primary emulsion;

[0042] (3) Add the required amount of peramivir to the primary emulsion, perform high-speed shearing (3000rpm-8000rpm)*(1min-5min), high-pressure homogenization (400bar-800bar)*(1min-5min), and spray dry the emulsion to obtain the final product.

[0043] In the preferred embodiment of the present invention, the high-speed shearing conditions in steps (1), (2), and (3) are (3000rpm-8000rpm)*(1min-5min).

[0044] In the preferred embodiment of the present invention, in step (2), the mass ratio of perfluorobromooctane to distearate phosphatidylcholine is 5-55:1, preferably 8-50:1, and more preferably 10-20:1.

[0045] In a preferred embodiment of the present invention, the solid content in the emulsion is 1%-4% by mass-volume percentage, preferably 1.5%-3.5%, and more preferably 2-3%.

[0046] In a preferred embodiment of the present invention, the inlet temperature of the spray dryer is 100-180°C, preferably 120-165°C.

[0047] In a preferred embodiment of the present invention, the emulsion temperature is maintained at 4-30°C during spray drying.

[0048] In a preferred embodiment of the present invention, the pharmaceutically acceptable carrier is a combination of phospholipids and calcium chloride.

[0049] In a preferred embodiment of the present invention, the phospholipid is selected from any one or a combination of distearylphosphatidylcholine (DSPC), dipalmitoylphosphatidylcholine, dilauroylphosphatidylcholine, dimyristoylphosphatidylcholine, hydrogenated soybean phospholipid, distearylphosphatidylglycerol, and dipalmitoylphosphatidylglycerol.

[0050] In a preferred embodiment of the present invention, the molar ratio of phospholipid to calcium chloride in the pharmaceutically acceptable carrier is 1:4-4:1, preferably 1-4:1.

[0051] In a preferred embodiment of the present invention, the composition contains 50-90% peramivir, 1-20% distearate phosphatidylcholine and 0.1-2% calcium chloride.

[0052] In a preferred embodiment of the present invention, the composition comprises 70-90% peramivir, 10-20% distearate phosphatidylcholine, and 0.5-2% calcium chloride.

[0053] In a preferred embodiment of the present invention, the aerodynamic diameter of the dry powder particles is 1-4.5 μm, preferably 1.5-4 μm, and more preferably 2.5-3.5 μm.

[0054] In a preferred embodiment of the present invention, the particle size distribution of the dry powder particles is D10≤0.5-2μm, D50≤2.5-4μm, and D90≤5-7.5μm.

[0055] In a preferred embodiment of the present invention, the particle size distribution of the dry powder particles is D10≤1-1.5μm, D50≤3-4μm, and D90≤5.5-7μm.

[0056] In the preferred embodiment of the present invention, the effective deposition rate (FPF) of the dry powder particles is ≥50%, preferably ≥60%, and more preferably ≥70%.

[0057] In a preferred embodiment of the present invention, the emptying rate (ER) of the dry powder particles is ≥90%, preferably ≥95%.

[0058] In a preferred embodiment of the present invention, the moisture content of the dry powder particles is ≤12%, preferably ≤10%.

[0059] In a preferred embodiment of the present invention, the surface of the dry powder particles has a porous structure.

[0060] In a preferred embodiment of the present invention, the geometric deviation (GSD) of the inhaled particles is 1-3, preferably 1.5-2.7.

[0061] In a preferred embodiment of the present invention, the surface of the dry powder particles has a porous structure.

[0062] In a preferred embodiment of the present invention, the dry powder inhaler delivers a pulmonary dose of at least about 10 mg in a single inhalation via pulmonary inhalation.

[0063] Another object of the present invention is to provide the use of the peramivir dry powder inhaler of the present invention in the preparation of a medicament for treating influenza virus.

[0064] In a preferred embodiment of the present invention, the influenza virus is any one or a combination of influenza A virus, influenza B virus, influenza C virus, and influenza D virus.

[0065] To clearly define the scope of protection of this invention, the following terms are defined:

[0066] Unless otherwise stated, this invention uses laser diffraction (NewPatek laser particle size analyzer) to determine particle size.

[0067] The "solids" mentioned in this invention refer to the total amount of non-volatile solids remaining after the material has been dried and evaporated to remove all moisture (and volatile substances) under specified conditions.

[0068] The "Mass Median Aerodynamic Diameter (MMAD)" mentioned in this invention refers to the aerodynamic diameter (μm) corresponding to a cumulative mass of 50% in an aerosol. This invention uses the multi-stage impactor method (NGI) to detect MMAD.

[0069] The "Fine Particle Fraction (FPF)" described in this invention refers to the percentage of the mass of particles with an aerodynamic diameter ≤5 μm in an aerosol (drug droplet / particle) relative to the total mass of the ejected / released drug. A higher FPF generally indicates better lung deposition. This invention uses a multi-stage impactor (NGI) to detect FPF.

[0070] This invention uses the emptying rate (ER) as an evaluation index to comprehensively characterize the flowability and atomization performance of tobramycin dry powder inhaler capsules. The emptying rate is calculated by measuring the mass change of the dry powder inhaler capsule before and after administration via an inhalation device and connection to a new generation impactor (NGI).

[0071] ER%=[(W2-W3) / (W2-W1)]*100%

[0072] W1: Capsule shell weight; W2: Capsule weight; W3: Capsule weight after inhalation.

[0073] This invention uses a Karl Fischer moisture analyzer (Metrohm 870 KF Titrino plus, Switzerland) to detect moisture content.

[0074] This invention uses LC / MS-MS to quantitatively detect the content of peramivir. The sample pretreatment method employs organic solvent precipitation of proteins: 20 μL of the drug-containing matrix is ​​placed in a 1.5 mL EP tube, and 80 μL of internal standard (IS) working solution (acetonitrile solution containing 5 ng / mL propranolol) is added. The mixture is vortexed for 1 min to mix, and then centrifuged at 10,000 rpm for 10 min at 4°C. 30 μL of the supernatant is added to 150 μL of 50% acetonitrile / water (v / v) solution, mixed, and then injected for analysis.

[0075] Liquid chromatography conditions: A C18 column was used, mobile phase A was 0.1% formic acid-water solution, mobile phase B was 0.1% formic acid-acetonitrile solution, gradient elution was used, the flow rate was 0.5 mL / min, the injection volume was 3 μL, and the total injection time was 3 min.

[0076] 0.0-0.3 min 95% 5% 0.3-1.0 min 95-40% 5-60% 1.0-2.0 min 40-5% 60-95% 2.0-2.2 min 5% 95% 2.2-2.21 min 5-95% 95-5% 2.21-3.0 min 95% 5%

[0077] Mass spectrometry conditions: positive ion detection, MRM scanning mode. The ion pairs used for analysis were peramivir (m / z = 329.4 / 270.4, DP = 120, CE = 30) and propranolol (m / z = 260.1 / 116.1, DP = 100, CE = 30). Mass spectrometry detection was performed using an electrospray ionization source in MRM mode, with the following parameters: ion spray voltage: +5500 V; capillary temperature: 550 °C; nebulizer gas: 50 psi; heating gas: 60 psi; curtain gas: 35 psi; collision gas: 8 psi; channel electron multiplier voltage: +1500 V.

[0078] Unless otherwise stated, when this invention relates to percentages between liquids, the percentage is volume / volume percentage; when this invention relates to percentages between liquids and solids, the percentage is volume / weight percentage; when this invention relates to percentages between solids and liquids, the percentage is weight / volume percentage; the remainder is weight / weight percentage.

[0079] Compared with the prior art, the present invention has the following beneficial effects:

[0080] 1. This invention scientifically screens the particle size, particle size distribution, appearance, and surface roughness of peramivir dry powder inhaler, precisely controls MMAD, achieves high effective deposition rate (FPF) and narrow particle distribution, realizes deep lung targeting, enhances efficacy and reduces toxicity, avoids systemic exposure risks and systemic toxicity, improves drug safety, efficacy, stability and patient compliance, and has the advantages of lung-targeted efficacy enhancement, systemic toxicity reduction, convenient operation, high antibacterial activity, long-term targeted effect, high safety and efficacy, good stability, high bioavailability and wide applicability.

[0081] 2. The preparation method of the peramivir dry powder inhaler of the present invention has the advantages of simple operation, lower cost, green and environmentally friendly, and suitable for industrial production. Detailed Implementation

[0082] The present invention is described in detail through the following examples and test cases, but is for illustrative purposes only and not intended to limit the scope of the invention. Peramivir was purchased from Ningbo Tianheng Pharmaceutical Co., Ltd., a subsidiary of Fu'an Pharmaceutical.

[0083] Example 1 Preparation of Peramivir Dry Powder Inhaler of the Present Invention

[0084] Composition of Peramivir dry powder inhaler:

[0085] Peramivir 1696.64 Distearate phosphatidylcholine 286.52 Calcium chloride 16.88

[0086] The preparation of peramivir dry powder inhaler includes the following steps:

[0087] (1) Add the required amount of distearylphosphatidylcholine and calcium chloride to 70℃ water (60mL), and shear at high speed (8000rpm*5min) to obtain an aqueous phase;

[0088] (2) Under stirring (8000 rpm), 2865 mg of perfluorooctane was added dropwise to the aqueous phase and the mixture was subjected to high-speed shearing at 8000 rpm for 5 min to obtain the primary emulsion.

[0089] (3) Add the required amount of peramivir to the primary emulsion, shear at high speed for 8000 rpm for 5 min, and then homogenize under high pressure (800 bar for 5 min) to obtain the emulsion (solid content 3%).

[0090] (4) Spray dry the emulsion (inlet temperature 135 ℃), keep the emulsion at 25℃, spray pressure flow rate 1052 L / h, and injection speed 6 mL / min to obtain the emulsion.

[0091] According to the method of this invention, the peramivir dry powder inhalation particles are porous spherical, and the geometric particle size distribution of the peramivir dry powder inhalation particles is: D 10 0.99μm, D 50 3.17μm, D 90 The particle size distribution of the peramivir dry powder inhaled particles was 6.17 μm; the water content was 8.982%; the fine particle fraction (FPF) was 62.458%; and the mass median aerodynamic diameter (MMAD) was 3.293 μm.

[0092] Example 2: Preparation of the Peramivir dry powder inhaler of the present invention

[0093] Composition of Peramivir dry powder inhaler:

[0094] Peramivir 1696.64 Distearate phosphatidylcholine 286.52 Calcium chloride 16.88

[0095] The preparation of peramivir dry powder inhaler includes the following steps:

[0096] (1) Add the required amount of distearylphosphatidylcholine and calcium chloride to 70℃ water (60mL), and shear at high speed (8000rpm*5min) to obtain an aqueous phase;

[0097] (2) Under stirring (8000 rpm), 5730 mg of perfluorooctane was added dropwise to the aqueous phase and the mixture was subjected to high-speed shearing at 8000 rpm for 5 min to obtain the primary emulsion.

[0098] (3) Add the required amount of peramivir to the primary emulsion, shear at high speed for 8000 rpm for 5 min, and then homogenize under high pressure (800 bar for 5 min) to obtain the emulsion (solid content 3%).

[0099] (4) Spray dry the emulsion (inlet temperature 135 ℃), keep the emulsion at 25℃, spray pressure flow rate 1052 L / h, and injection speed 6 mL / min to obtain the emulsion.

[0100] According to the method of this invention, the peramivir dry powder inhalation particles are porous spherical, and the geometric particle size distribution of the peramivir dry powder inhalation particles is: D 10 1.09μm, D 50 3.26μm, D 90The particle size distribution of the peramivir dry powder inhaled particles was 6.63 μm; the water content was 11.657%; the fine particle fraction (FPF) was 59.078%; and the mass median aerodynamic diameter (MMAD) was 3.477 μm.

[0101] Example 3: Preparation of the Peramivir dry powder inhaler of the present invention

[0102] Composition of Peramivir dry powder inhaler:

[0103] Peramivir 2544.96 Distearate phosphatidylcholine 429.6 Calcium chloride 50.64

[0104] The preparation of peramivir dry powder inhaler includes the following steps:

[0105] (1) Add the required amount of distearylphosphatidylcholine and calcium chloride to 70℃ water (90mL), and shear at high speed (8000rpm*5min) to obtain an aqueous phase;

[0106] (2) Under stirring (8000 rpm), 4297.5 mg of perfluorooctane was added dropwise to the aqueous phase and the mixture was subjected to high-speed shearing at 8000 rpm for 5 min to obtain the primary emulsion.

[0107] (3) Add the required amount of peramivir to the primary emulsion, shear at high speed for 8000 rpm for 5 min, and then homogenize under high pressure (800 bar for 5 min) to obtain the emulsion (solid content 3%).

[0108] (4) Spray dry the emulsion (inlet temperature 135 ℃), keep the emulsion at 25℃, spray pressure flow rate 1052 L / h, and injection speed 6 mL / min to obtain the emulsion.

[0109] According to the method of this invention, the peramivir dry powder inhalation particles are porous spherical, and the geometric particle size distribution of the peramivir dry powder inhalation particles is: D 10 1.08μm, D 50 3.48μm, D 90 The particle size distribution of the peramivir dry powder inhaled particles was 6.25 μm; the water content was 9.453%; the fine particle fraction (FPF) was 73.785%; and the mass median aerodynamic diameter (MMAD) was 1.871 μm.

[0110] Example 4: Preparation of the Peramivir dry powder inhaler of the present invention

[0111] Composition of Peramivir dry powder inhaler:

[0112] Peramivir 2544.96 Distearate phosphatidylcholine 429.6 Calcium chloride 25.32

[0113] The preparation of peramivir dry powder inhaler includes the following steps:

[0114] (1) Add the required amount of distearylphosphatidylcholine and calcium chloride to 70℃ water (74mL), and shear at high speed (8000rpm*5min) to obtain an aqueous phase;

[0115] (2) Under stirring (8000 rpm), 4297.5 mg of perfluorooctane was added dropwise to the aqueous phase and the mixture was subjected to high-speed shearing at 8000 rpm for 5 min to obtain the primary emulsion.

[0116] (3) Add the required amount of peramivir to the primary emulsion, shear at high speed for 8000 rpm for 5 min, and then homogenize under high pressure (800 bar for 13 min) to obtain the emulsion (solid content 4%).

[0117] (4) Spray dry the emulsion (inlet temperature 135℃), keep the emulsion at 25℃, spray pressure flow rate 1052 L / h, and injection speed 6 mL / min to obtain the emulsion.

[0118] According to the method of this invention, the peramivir dry powder inhalation particles are porous spherical, and the geometric particle size distribution of the peramivir dry powder inhalation particles is: D 10 1.24μm, D 50 3.63μm, D 90 The particle size distribution of the peramivir dry powder inhaled particles was 6.72 μm; the water content was 11.342%; the fine particle fraction (FPF) was 53.96%; and the mass median aerodynamic diameter (MMAD) was 3.927 μm.

[0119] Example 5: Preparation of the Peramivir dry powder inhaler of the present invention

[0120] Composition of Peramivir dry powder inhaler:

[0121] Peramivir 2544.96 Distearate phosphatidylcholine 429.6 Calcium chloride 25.32

[0122] The preparation of peramivir dry powder inhaler includes the following steps:

[0123] (1) Add the required amount of distearylphosphatidylcholine and calcium chloride to 70℃ water (124mL), and shear at high speed (8000rpm*5min) to obtain an aqueous phase;

[0124] (2) Under stirring (8000 rpm), 4297.5 mg of perfluorooctane was added dropwise to the aqueous phase and subjected to high-speed shearing at 8000 rpm for 5 min to obtain the primary emulsion (solid content 2%).

[0125] (3) Add the required amount of peramivir to the primary emulsion, shear at high speed for 8000 rpm for 5 min, and then homogenize under high pressure (800 bar for 15 min) to obtain the emulsion;

[0126] (4) Spray dry the emulsion (inlet temperature 135℃), keep the emulsion at 25℃, spray pressure flow rate 1052 L / h, and injection speed 6 mL / min to obtain the emulsion.

[0127] The geometric particle size distribution of peramivir dry powder inhaled particles was detected according to the method of this invention: D 10 1.17μm, D 50 3.21μm, D 90 The particle size distribution of the peramivir dry powder inhaled particles was 6.09 μm; the water content was 9.982%; the fine particle fraction (FPF) was 59.951%; and the mass median aerodynamic diameter (MMAD) was 3.514 μm.

[0128] Example 6: Preparation of the Peramivir dry powder inhaler of the present invention

[0129] Composition of Peramivir dry powder inhaler:

[0130] Peramivir 1944.96 Distearate phosphatidylcholine 429.6 Calcium chloride 25.32

[0131] The preparation of peramivir dry powder inhaler includes the following steps:

[0132] (1) Add the required amount of distearylphosphatidylcholine and calcium chloride to 70℃ water (72mL), and shear at high speed (8000rpm*5min) to obtain an aqueous phase;

[0133] (2) Under stirring (8000 rpm), 4297.5 mg of perfluorooctane was added dropwise to the aqueous phase and the mixture was subjected to high-speed shearing at 8000 rpm for 5 min to obtain the primary emulsion.

[0134] (3) Add the required amount of peramivir to the primary emulsion, shear at high speed for 8000 rpm for 5 min, and then homogenize under high pressure (800 bar for 10 min) to obtain the emulsion (solid content 3%).

[0135] (4) Spray dry the emulsion (inlet temperature 135℃), keep the emulsion at 25℃, spray pressure flow rate 1052 L / h, and injection speed 6 mL / min to obtain the emulsion.

[0136] The geometric particle size distribution of peramivir dry powder inhaled particles was detected according to the method of this invention: D 10 1.13μm, D 50 3.32μm, D 90 The particle size distribution of the peramivir dry powder inhaled particles was 5.97 μm; the water content was 8.257%; the fine particle fraction (FPF) was 57.275%; and the mass median aerodynamic diameter (MMAD) was 3.770 μm.

[0137] Experimental Example 1: Pharmacokinetic Study of the Peramivir Dry Powder Inhaler of the Present Invention

[0138] Twelve male SD rats (purchased from Spefair) weighing 200-230g were randomly divided into two groups of six each:

[0139] Intravenous administration group: Experimental animals were injected via tail vein with peramivir solution (250 mg peramivir dissolved in 50 mL of physiological saline to prepare a peramivir solution with a concentration of 5 mg / mL), the dosage was 6 mg / kg, and the administration volume was 5 mL / kg;

[0140] Inhalation group: Using a lung dry powder delivery device, the experimental animals were given the dry powder inhaler of Example 3 via trachea at a dose of 6 mg / kg and a volume of 200 μL / animal.

[0141] Following drug administration, blood samples (approximately 100 μL each time) were collected from the jugular vein of the experimental animals before administration and at 0.033 h, 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, 24 h, and 36 h after administration. The samples were placed in heparin sodium anticoagulant tubes and centrifuged at 2500 × g for 10 min at 4 °C. The supernatant plasma was collected and stored at -40 °C for later analysis. The sample processing method was the organic solvent precipitation method for proteins.

[0142] The blood concentration of peramivir was determined using the method of this invention, and pharmacokinetic parameters were calculated using noncompartmental analysis (NCA). The results are shown in Table 1. Peramivir does not tend to distribute into tissues, indicating that pulmonary inhalation can increase pulmonary drug exposure and reduce systemic drug exposure.

[0143] Table 1 Pharmacokinetic parameters (n=6, mean±s)

[0144] <![CDATA[T max (h)]]> 0.60±0.40 / <![CDATA[C max (ng / mL)]]> 2225±398.1 22732±5543 <![CDATA[AUC las (h×ng / mL)]]> 6641.8±859.9 6048±636 <![CDATA[T 1 / 2 (h)]]> 3.1±1.3 3.07±0.61 <![CDATA[MRT las (h)]]> 1.6±0.3 0.47±0.06

[0145] The above description of specific embodiments of the present invention does not limit the present invention. Those skilled in the art can make various changes or modifications based on the present invention, and as long as they do not depart from the spirit of the present invention, they should all fall within the scope of protection of the claims of the present invention.

Claims

1. A peramivir dry powder inhaler, wherein, by weight percentage, the dry powder inhaler comprises 50-90% peramivir and a pharmaceutically acceptable carrier, wherein, The aerodynamic diameter (MMAD) of the dry powder particles that make up the dry powder inhaler is 1μm-5μm, and the particle size distribution of the dry powder particles is D10≤2.0μm, D50≤4.0μm, and D90≤7.5μm.

2. The dry powder inhaler of claim 1, wherein the pharmaceutically acceptable carrier is a combination of phospholipids and calcium chloride.

3. The dry powder inhaler according to any one of claims 1-2, wherein the phospholipid is selected from any one or a combination of distearylphosphatidylcholine (DSPC), dipalmitoylphosphatidylcholine, dilauroylphosphatidylcholine, dimyristoylphosphatidylcholine, hydrogenated soybean phospholipid, distearylphosphatidylglycerol, and dipalmitoylphosphatidylglycerol.

4. The dry powder inhaler according to any one of claims 1-3, wherein the molar ratio of phospholipid to calcium chloride in the pharmaceutically acceptable carrier is 1:4-4:1, preferably 1-4:

1.

5. The dry powder inhaler according to any one of claims 1-4, wherein the dry powder inhaler contains 50-90% peramivir, 1-20% distearate phosphatidylcholine and 0.1-2% calcium chloride.

6. The dry powder inhaler according to any one of claims 1-5, wherein the dry powder inhaler contains 70-90% peramivir, 10-20% distearate phosphatidylcholine and 0.5-2% calcium chloride.

7. The dry powder inhaler according to any one of claims 1-6, wherein the aerodynamic diameter of the dry powder particles is 1-4.5 μm, preferably 1.5-4 μm, and more preferably 2.5-3.5 μm.

8. The dry powder inhaler according to any one of claims 1-7, wherein the effective deposition rate (FPF) of the dry powder particles is ≥50%, preferably ≥60%, and more preferably ≥70%.

9. The method for preparing the peramivir dry powder inhaler according to any one of claims 1-8, wherein the preparation of the peramivir dry powder inhaler comprises the following steps: (1) Add the required amount of distearate phosphatidylcholine and calcium chloride to water at 70-80℃, and shear at high speed (3000rpm-8000rpm)* (1min-5min) to obtain an aqueous phase; (2) Under stirring conditions (3000rpm-8000rpm), the required amount of perfluorooctane (PFOB) was added to the aqueous phase and subjected to high-speed shearing (3000rpm-8000rpm)* (1min-5min) to obtain the primary emulsion; (3) Add the required amount of peramivir to the primary emulsion, perform high-speed shearing (3000rpm-8000rpm)*(1min-5min), high-pressure homogenization (400bar-800bar)*(1min-5min), and spray dry the emulsion to obtain the final product.

10. The use of the peramivir dry powder inhaler as described in any one of claims 1-8 in the preparation of a medicament for treating influenza virus.