A dry powder inhaler for an efficient and broad-spectrum anti-coronavirus polypeptide and a preparation method thereof

Abstract

The present invention belongs to the field of preparations of polypeptide drugs, and relates to a dry powder inhaler for an anti-coronavirus polypeptide with high efficiency and broad spectrum and a preparation method thereof. When the dry powder inhaler is used for pulmonary administration through a dry powder inhalation device, it has excellent inhalation characteristics, delivery effect and stability effect, and has the characteristics of high bioavailability, convenient to carry, convenient to administer and pollution-free; moreover, the preparation method is simple, suitable for industrial production, and has good application value.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a dry powder inhaler for an anti-coronavirus polypeptide with high efficiency and broad spectrum and a preparation method thereof. Background Art

[0002] It has been generally recognized that pulmonary drug delivery is not only an effective therapy for treating local pulmonary diseases (such as asthma, COPD, bronchiectasis, pulmonary infections, etc.), but also a commonly used therapy for obtaining systemic therapeutic effects. After inhaling drugs into the lungs, the drugs directly reach the lungs. Compared with oral administration, the drug dosage can be significantly reduced, thereby reducing drug adverse reactions. The uniqueness of pulmonary anatomy and physiology is one of the key determinants of inhaled drugs. The lungs are rich in capillaries, 80% of the alveoli are wrapped with capillaries, and the blood flow in the blood vessels is large. After the drugs are absorbed through the pulmonary blood vessels, they directly enter the blood without a first-pass effect. Therefore, the absorption after pulmonary drug delivery is very rapid. The biotransformation enzymes in the lungs are concentrated in type II alveolar cells, which can reduce the extensive hydrolysis of proteins and polypeptides to a certain extent after they enter the lungs, enabling the drugs to maintain their biological activities and achieve the drug effects. Pulmonary drug delivery preparations are mainly divided into three types: nebulizers, pressurized metered-dose inhalers, and dry powder inhalers. Each of these three types of drug delivery preparations has its own characteristics. Pressurized metered-dose inhalers and nebulizers use propellants to provide power for drug delivery, which is convenient to use. However, the propellant Freon used in both of them will pollute the environment, and the pressurized metered-dose inhalers cannot deliver large doses of drugs, which limits the further development of these two dosage forms.

[0003] A dry powder inhaler (DPI), also known as an inhaled powder aerosol, refers to a preparation in which a solid micronized active pharmaceutical ingredient alone or in combination with a suitable carrier is in the form of a capsule, blister, or multi-dose reservoir, and is inhaled into the lungs by the patient actively using a special dry powder inhalation device. It has many characteristics such as easy to use, environmentally friendly, without propellants, high inhalation efficiency, good drug stability, and non-invasive drug delivery. The power of the dry powder inhaler comes from the patient's active inhalation, which solves the above environmental problems. Moreover, because it can deliver large doses of drugs and the drugs are solid powders, it can be used to deliver proteins and peptide drugs, expanding the application scope of pulmonary drug delivery preparations.

[0004] YKYY017 is a broad-spectrum and highly effective coronavirus membrane fusion inhibitor with antiviral activity. It is a polypeptide drug that can be used for the prevention and treatment of COVID-19. This product is a lipopeptide compound composed of 43 amino acids. YKYY017 forms a heterologous six-helix bundle (6-HB) by interacting with the heptapeptide repeat region 1 (HR1) of the S2 subunit of the SARS-CoV-2 spike protein, thereby inhibiting the formation of the homologous 6-HB between the HR1 and HR2 domains of the virus itself and blocking the fusion process between the virus and host cells, thus achieving the purpose of antiviral. In vitro pharmacodynamic studies have shown that YKYY017 has significant inhibitory effects on the original strain of the novel coronavirus and its various prevalent variants (Delta, Omicron BA.1, Omicron BA.2, Omicron BA.4, and Omicron BF.7), and is not affected by virus mutations. In vivo pharmacodynamic studies have shown that after treatment with YKYY017 after virus challenge, it can significantly reduce the viral load in the lungs of hamsters infected with SARS-CoV-2 (Delta and Omicron BA.2 variants) and effectively relieve the pathological damage in the lungs of infected hamsters. The results of toxicology studies have shown that YKYY017 has no obvious toxicity and related adverse reactions, and has good safety.

[0005] In order to solve the problems of slow onset, low bioavailability, environmental pollution, or inconvenient use of some pharmaceutical products, there is an urgent need to provide a dry powder inhaler for COVID-19 polypeptides with good inhalation characteristics, delivery effect, and stability. Summary of the Invention

[0006] The present invention mainly provides a novel preparation of a drug for preventing or treating coronavirus infection, and provides a dry powder inhaler for highly effective broad-spectrum anti-coronavirus polypeptides and its preparation method.

[0007] The dry powder inhaler prepared by the method of the present invention can be absorbed through the lungs, reduce the virus in the body of mice infected with the novel coronavirus, and improve the symptoms. The freeze-dried powder of the preparation has good fluidity (angle of repose < 45°), good inhalation characteristics (aerodynamic mass median diameter is small, as low as 2.59 μm; fine particle dose content is as high as 42.6%), excellent delivery effect (delivery dose is as high as 97%, delivery dose uniformity is good, distribution is narrow, only 91% - 110%), and good stability (the change rate of the content of the drug active ingredient is as low as 0.1%).

[0008] The present invention mainly provides a dry powder inhaler, wherein the drug active ingredient includes polypeptides.

[0009] In some preferred embodiments, the polypeptide includes 20 - 60 amino acids.

[0010] In some preferred embodiments, the polypeptide is used for treating infections caused by coronaviruses.

[0011] In some more preferred embodiments, the polypeptide comprises YKYY017, and the peptide sequence of YKYY017 is shown as SEQ ID NO.1:

[0012] Ac-Ser-Val-Val-Asn-Ile-Gln-Lys-Glu-Ile-Asp-Arg-Leu-Asn-Glu-Val-Ala-Lys-Asn-Leu-Asn-Glu-Ser-Leu-Ile-Asp-Leu-Gln-Glu-Leu-Gly-Lys-Tyr-Glu-Gln-Tyr-Ile-Lys-Glu-Ala-Ala-Ala-Lys-Lys(Chol)-NH 2 。

[0013] Among them, the acetyl group (Ac-) is an amino-terminal protecting group; the amino group (-NH 2 ) is a carboxyl-terminal protecting group; Chol is a cholesteryl succinate monoester group modified on the C-terminal amino acid lysine (Lys).

[0014] In some preferred embodiments, the dry powder inhaler further comprises any one or more of the following components: stabilizers, excipients, carriers.

[0015] In some preferred embodiments, the stabilizer is a buffered saline solution with a pH of 3.0 to 4.0.

[0016] In some more preferred embodiments, the pH of the stabilizer is 3.7.

[0017] In some preferred embodiments, the stabilizer comprises: acetic acid - acetate solution and / or citric acid - citrate solution.

[0018] In some more preferred embodiments, the dry powder inhaler comprises a pharmaceutically active ingredient, a stabilizer, an excipient and a carrier; the stabilizer comprises: a citric acid - citrate solution with a pH of 3.0 to 4.0.

[0019] In some preferred embodiments, the excipient is selected from one or more of the following substances: mannitol, sucrose, trehalose.

[0020] In some preferred embodiments, the carrier is selected from one or more of the following substances: lactose, mannitol, erythritol, xylitol, gum arabic; preferably lactose and / or mannitol.

[0021] In some preferred embodiments, the dosage of the carrier is 15 to 50 parts by mass, preferably, the dosage of the carrier is 30 to 35 parts by mass.

[0022] In some more preferred embodiments, the dry powder inhalant further comprises an anticoagulant.

[0023] In some preferred embodiments, the anticoagulant is selected from one or more of the following substances: leucine, mannitol, poloxamer, phospholipid; preferably leucine and / or phospholipid.

[0024] In some preferred embodiments, the dosage of the drug active ingredient is 5 to 15 parts by mass.

[0025] In some preferred embodiments, the dosage of the anticoagulant is 5 to 18 parts by mass, preferably, the dosage of the anticoagulant is 8 to 12 parts by mass.

[0026] In some preferred embodiments, the dosage of the excipient is 2 to 8 parts by mass.

[0027] The present invention also provides a method for synthesizing the polypeptide as follows:

[0028] a) Select a resin and couple all amino acids in sequence from the C-terminus to the N-terminus to synthesize a fully protected polypeptide by solid-phase synthesis; preferably use Rink Amide-MBHA Resin;

[0029] b) Remove the side-chain protecting group of Lys; preferably use hydrazine as a catalyst to selectively remove the side-chain protecting group;

[0030] c) Couple cholesterol succinate monoester to the side chain in the presence of a coupling agent;

[0031] d) In the presence of a cleavage agent, cleave the peptide resin and deprotect the side chain to obtain the polypeptide.

[0032] In some preferred embodiments, the dry powder inhalant is administered by oral inhalation using a Turbohaler.

[0033] The present invention also provides a method for preparing the dry powder inhalant, comprising the following steps:

[0034] ① Prepare a stabilizer;

[0035] ② Dissolve the excipient in the stabilizer;

[0036] ③ Dissolve the drug active ingredient in the system of step ②, adjust the pH to obtain a solution;

[0037] ④ Lyophilize the solution obtained in step ③ to obtain a lyophilized powder;

[0038] ⑤ Crushing;

[0039] ⑥ Mixing: Mix the crushed powder with a carrier, and then crush to obtain a dry powder inhalant.

[0040] In some preferred embodiments, it further includes mixing an anticoagulant with the lyophilized powder and crushing in step ⑤.

[0041] In some preferred embodiments, in step ⑤, the particle size D90 after crushing is below 10 μm, preferably 1.0 μm to 5.0 μm.

[0042] In some preferred embodiments, in step ⑥, the carrier particle size is between 1 μm and 250 μm, wherein D50 is 80 μm to 120 μm.

[0043] In some preferred embodiments, after lyophilization in step ④, the moisture content is controlled to be ≤ 1.0%.

[0044] The present invention also provides the use of a dry powder inhalant in the preparation of a drug for treating coronavirus infection.

[0045] The beneficial effects of the present invention are as follows:

[0046] I. Stabilizer

[0047] 1. Using a buffer system as a stabilizer (for example, acetic acid - acetate solution, citric acid - citrate buffer solution) to prepare a dry powder inhalant, the inhalation characteristics and delivery effect of the resulting preparation are good, and the stability is remarkable. Specifically, it is shown as follows:

[0048] i. The median aerodynamic mass diameter of the drug in the resulting preparation is small, only 3.04 μm to 3.59 μm, less than 5 μm (particles that can enter the lungs are below 5 μm); and the dose of fine drug particles is large, 25.6% to 30.1%, far higher than the pharmacopoeia requirement (≥ 10%), indicating that there are sufficient drug particles to enter the lungs;

[0049] ii. The delivery dose of the resulting preparation is high and the delivery dose uniformity is good, with the delivery dose being 88% to 89%; the delivery dose uniformity distribution is between 77% and 117%, and the distribution range is narrow, higher than the limit standard (75% to 125%);

[0050] iii. The resulting preparation has good stability. After the accelerated test, the change rate of the content of the drug active ingredient is low, only 0.1% to 0.5%, far less than 5%.

[0051] 2. Using other solutions or solutions without buffer capacity as stabilizers (for example, hydrochloric acid solution) to prepare a dry powder inhalant, the drug stability is poor and it cannot be used as a qualified preparation product. Specifically, it is shown as follows:

[0052] The obtained preparation has poor stability. After the accelerated test, the change rate of the content of the drug active ingredient is significant, being 14.2%, far greater than the specified standard (≤5%), and it cannot be used as a qualified dry powder inhaler.

[0053] II. pH of the stabilizer

[0054] 1. When using a buffer system with a pH of 3.0 - 4.0 as the stabilizer to prepare a dry powder inhaler, the obtained preparation has good inhalation characteristics and delivery effect, and good and significant stability, specifically manifested as:

[0055] i. The median aerodynamic mass diameter of the drug in the obtained preparation is small, being 2.69 μm - 3.26 μm, less than 5 μm; the fine particle dose of the drug in the obtained preparation is large, being 25.5% - 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles that can enter the lungs;

[0056] ii. The obtained preparation has a high delivery dose and good delivery dose uniformity. The delivery dose is 85% - 89%; the delivery dose uniformity is distributed between 79% - 119%, with a narrow distribution range, far higher than the limit standard (75% - 125%);

[0057] iii. The obtained preparation has good stability. After the accelerated experiment, the change rate of the content of the drug active ingredient is small, being 0.5% - 1.9%, less than 5%.

[0058] 2. When using a buffer system with a pH of 3.7 as the stabilizer, the obtained preparation has the best inhalation characteristics and delivery effect, and the optimal stability; specifically manifested as:

[0059] i. The median aerodynamic mass diameter of the drug in the obtained preparation is small, being 3.04 μm, less than 5 μm;

[0060] The fine particle dose of the drug in the obtained preparation is large, being 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles that can enter the lungs;

[0061] ii. The obtained preparation has a high delivery dose and good delivery dose uniformity. The delivery dose is 89%; the delivery dose uniformity is distributed between 80% - 116%, with a narrow distribution range, far higher than the limit standard of 75% - 125%;

[0062] iii. The obtained preparation has the optimal stability. After the accelerated test, the change rate of the content of the drug active ingredient is small, being 0.5%, far less than 5%, and there is basically no change.

[0063] 3. When the pH value of the buffer system is less than 3.0 or greater than 4.0, the resulting preparation has poor stability and cannot be used as a qualified preparation product. Specifically, it is manifested as follows:

[0064] The resulting preparation has poor stability. After the accelerated test, the change rate of the content of the drug active ingredient is large, ranging from 10.0% to 15.6%, far greater than 5%, showing a significant change, and it cannot be used as a qualified dry powder inhaler.

[0065] III. Types of Excipients

[0066] 1. The dry powder inhaler prepared by using the excipients of the present invention (such as mannitol, trehalose, sucrose) has good fluidity of the obtained freeze-dried powder, and excellent inhalation characteristics and delivery effects. Specifically, it is manifested as follows:

[0067] i. Good fluidity, with a small angle of repose of 50.2° - 58.1°; the bulk density is 0.13 g / cm 3 - 0.17 g / cm 3 ; small particles, with a particle size D90 of 2.52 μm - 3.12 μm; having good fluidity;

[0068] ii. The drug aerodynamic mass median diameter of the resulting preparation is small, ranging from 3.04 μm to 4.96 μm, less than 5 μm; the fine particle dose of the drug in the resulting preparation is large, ranging from 19.5% to 30.1%, greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles that can enter the lungs;

[0069] iii. The resulting preparation has a high delivery dose and good delivery dose uniformity, with a delivery dose of 84% - 90%; the delivery dose uniformity is distributed within 75% - 125%.

[0070] 2. When mannitol is used as the excipient, the resulting freeze-dried powder has excellent fluidity, and the inhalation characteristics and delivery effects of the preparation are the best. Specifically, it is manifested as follows:

[0071] i. The best fluidity, with an angle of repose as low as 50.2°; the bulk density is 0.17 g / cm 3 ; the particle size D90 is 2.93 μm;

[0072] ii. The drug aerodynamic mass median diameter of the resulting preparation is small, being 3.04 μm, less than 5 μm; the fine particle dose of the drug in the resulting preparation is large, being 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles that can enter the lungs;

[0073] iii. The delivery dose of the resulting preparation is high and the delivery dose uniformity is good, with the delivery dose being 89%; the delivery dose uniformity is distributed between 80% and 116%, with a narrow distribution range, far higher than the limit standard of 75% - 125%.

[0074] IV. Types of carriers

[0075] 1. The dry powder inhalant prepared with the carrier of the present invention (such as lactose, mannitol, erythritol, xylitol, gum arabic) has good hygroscopicity, and excellent inhalation characteristics, delivery effect and stability. Specifically manifested as:

[0076] i. The resulting preparation has good hygroscopicity, and the moisture absorption rate is controlled within 0.3% - 3.8%, less than 5%, with low hygroscopicity;

[0077] ii. The resulting preparation has a small aerodynamic mass median diameter of the drug, which is 3.04 μm - 4.63 μm, less than 5 μm; the fine particle dose of the drug in the resulting preparation is large, which is 21.6% - 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles that can enter the lungs;

[0078] iii. The resulting preparation has a high delivery dose and good delivery dose uniformity, with the delivery dose being 82% - 90%; the delivery dose uniformity is distributed between 75% and 124%, with a narrow distribution range;

[0079] iv. The resulting preparation has good stability. After the accelerated test, the change rate of the fine particle dose of the drug is controlled within 1.8% - 4.2%, less than 5%.

[0080] 2. The dry powder inhalant prepared with one of lactose and mannitol as the carrier has the lowest hygroscopicity, and excellent inhalation characteristics, delivery effect and the best stability. Specifically manifested as:

[0081] i. The resulting preparation has good hygroscopicity, and the moisture absorption rate is controlled within 0.3% - 0.5%, significantly lower than 5%,

[0082] with the best hygroscopicity;

[0083] ii. The resulting preparation has a small aerodynamic mass median diameter of the drug, which is 3.04 μm - 3.56 μm, less than 5 μm; the fine particle dose of the drug in the resulting preparation is large, which is 27.5% - 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles that can enter the lungs;

[0084] iii. The resulting preparation has a high delivery dose and good delivery dose uniformity, with the delivery dose being 83% - 89%; the delivery dose uniformity is distributed between 80% and 124%, with a narrow distribution range;

[0085] iv. The stability of the obtained preparation is optimal. After the accelerated test, the change rate of the dose of the fine drug particles is only 1.8% - 2.2%, far less than 5%.

[0086] V. Carrier dosage

[0087] 1. For the dry powder inhalant prepared by adding 20 - 50 parts of the carrier, the powder fluidity, inhalation characteristics and delivery effect of the obtained preparation are good. Specifically manifested as:

[0088] i. The angle of repose of the obtained preparation is small, controlled within 36.4° - 45.5°, and the material fluidity is good;

[0089] ii. The aerodynamic mass median diameter of the drug in the obtained preparation is small, being 3.04μm - 4.54μm, less than 5μm; the dose of the fine drug particles in the obtained preparation is large, being 23.3% - 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles to enter the lungs;

[0090] iii. The obtained preparation has a high delivery dose and good delivery dose uniformity. The delivery dose is 83% - 89%; the delivery dose uniformity is distributed between 80% - 119%, with a narrow distribution range, far higher than the limit standard of 75% - 125%.

[0091] 2. For the dry powder inhalant prepared by using 30 - 35 parts of the carrier, the inhalation characteristics of the obtained preparation are the best. Specifically manifested as:

[0092] i. The angle of repose of the obtained preparation is relatively small, being 40.2°, and the material fluidity is good;

[0093] ii. The aerodynamic mass median diameter of the drug in the obtained preparation is small, being 3.04μm, less than 5μm;

[0094] The dose of the fine drug particles in the obtained preparation is large, being 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles to enter the lungs;

[0095] iii. The obtained preparation has a high delivery dose and good delivery dose uniformity. The delivery dose is 89%; the delivery dose uniformity is distributed between 80% - 116%, with a narrow distribution range, far higher than the limit standard of 75% - 125%.

[0096] VI. Anticoagulant

[0097] 1. During the preparation process of the polypeptide dry powder inhalant, compared with not adding an anticoagulant, after adding an anticoagulant, the powder fluidity, inhalation characteristics, delivery effect and stability of the prepared preparation are the best. Specifically manifested as:

[0098] i. The angle of repose of the obtained freeze-dried powder is the smallest, as low as 38.2°, and it has the best fluidity.

[0099] ii. The aerodynamic mass median diameter of the drug in the obtained preparation is small, being 3.04 μm, less than 5 μm.

[0100] And it is reduced by 0.45 μm compared with that without anticoagulant; the fine particle dose of the drug in the obtained preparation is large, being 42.6%, far greater than the specified standard of 10%, and it is increased by 12.5% compared with that without anticoagulant; it shows better inhalation characteristics.

[0101] iii. The delivery dose of the obtained preparation is high and the uniformity is good. The delivery dose range is 91% - 110%, and the average delivery dose is 97% of the labeled amount, which is significantly improved compared with that without adding anticoagulant (89%).

[0102] iv. The obtained preparation has better stability. After the accelerated test, the change rate of the fine particle dose of the drug is only 1.1%, less than 5%; and it is further reduced by 0.7% compared with that without anticoagulant.

[0103] VII. Types of Anticoagulants

[0104] 1. By using the anticoagulants of the present invention (for example, leucine, mannitol, poloxamer, and phospholipid), dry powder inhalants with good fluidity, inhalation characteristics, and delivery effects can be prepared. Specifically manifested as:

[0105] i. The angle of repose of the obtained dry powder inhalant is small and the fluidity is good.

[0106] ii. The aerodynamic mass median diameter of the drug in the obtained preparation is 2.59 μm - 3.77 μm, less than 5 μm.

[0107] The fine particle dose of the drug in the obtained preparation is 37.6% - 42.6%, far greater than 10%; the inhalation characteristics are good.

[0108] iii. The delivery dose of the obtained preparation is 90% - 98%, meeting the limit standard (80% - 120%), and the delivery dose is high; the uniformity range of the delivery dose is 80% - 121%, and the uniformity is good.

[0109] 2. When the anticoagulants are leucine and phospholipid, the fluidity, inhalation characteristics, and delivery effects of the obtained preparation are better. Specifically manifested as:

[0110] i. The angle of repose of the obtained dry powder inhalant is relatively small and the fluidity is relatively good.

[0111] ii. The aerodynamic mass median diameter of the drug in the obtained preparation is relatively small, being 2.59 μm and 2.89 μm respectively, less than 5 μm; the fine particle dose of the drug in the obtained preparation is relatively large, being 42.6% respectively

[0112] and 38.7%, far greater than 10%; good inhalation characteristics;

[0113] iii. The delivery doses of the obtained preparations are 97% and 98% respectively, meeting the limit standards (80% - 120%), with high delivery doses; the delivery dose uniformity ranges are 91% - 110% and 92% - 113% respectively, with good uniformity. Detailed implementation manners

[0114] The present invention will be further illustrated by the following examples. It should be understood that: the examples of the present invention are only given for the purpose of illustrating the present invention, rather than limiting the present invention. Any simple improvement to the present invention under the premise of the technical solution of the present invention belongs to the protection scope of the present invention.

[0115] Example 1:

[0116] Prescription:

[0117] Table 1: Component table of Example 1

[0118]

[0119]

[0120] Preparation method:

[0121] Prepare according to the prescription ingredients and dosages shown in the above table;

[0122] Step ① Prepare an acetic acid - sodium acetate buffer solution with pH 3.7: Add sodium acetate and acetic acid to injection water, stir and dissolve to obtain an acetic acid - sodium acetate buffer solution;

[0123] Step ② Dissolve mannitol in the above acetic acid - sodium acetate buffer solution with pH 3.7, stir evenly to obtain a mannitol solution;

[0124] Step ③ Add YKYY017 to the above mannitol solution, stir and dissolve, adjust the pH value to 3.7 to obtain a YKYY017 solution;

[0125] Step ④ Freeze - dry the above YKYY017 solution, control the water content < 1.0% to obtain YKYY017 freeze - dried powder;

[0126] Step ⑤ Crush the above YKYY017 freeze - dried powder with a fluid energy mill, control the powder particle size, D90 is 1.0 μm - 5.0 μm to obtain YKYY017 freeze - dried powder;

[0127] Step ⑥ Add the above-mentioned YKYY017 lyophilized powder and lactose into a high-speed stirring mixer, start stirring at 120 rpm, cutting at 2000 rpm, and mix for 10 minutes. After mixing, obtain the YKYY017 dry powder inhaler;

[0128] During the application process, a blister-type or capsule-type Accuhaler drug delivery system is used for drug delivery.

[0129] Example 2: Different types of stabilizers (buffer solutions)

[0130] On the basis of Example 1, in order to explore the influence of the type of buffer system on the YKYY017 dry powder inhaler, change the type of buffer system as the stabilizer in the prescription, keep the prescription dosage unchanged, and on the basis of controlling the pH of the buffer solution at 3.7, replace the acetic acid-sodium acetate buffer solution with:

[0131] Method 1 Citric acid-sodium citrate buffer solution

[0132] Method 2 Hydrochloric acid solution.

[0133] The specific components and dosages are shown in the following table:

[0134] Table 2: Component table of Method 1 in Example 2

[0135]

[0136]

[0137] Table 3: Component table of Method 2 in Example 2

[0138]

[0139] Preparation method:

[0140] Except for steps ① and ③, the other preparation methods are the same as those in Example 1.

[0141] Method 1:

[0142] Step ① Prepare a citric acid-sodium citrate buffer solution with a pH of 3.7: Add sodium citrate and citric acid to injection water, stir and dissolve to obtain a citric acid-sodium citrate buffer solution;

[0143] Step ③ Add YKYY017 to the above-mentioned mannitol solution, stir and dissolve, and adjust the pH value to 3.7 to obtain a YKYY017 solution.

[0144] Method 2:

[0145] Step ① Prepare a hydrochloric acid solution with a pH of 3.7: Add hydrochloric acid to injection water, stir evenly to obtain a hydrochloric acid buffer solution;

[0146] Step ③: Add YKYY017 to the above-mentioned mannitol solution, stir to dissolve, and adjust the pH value to 3.7 to obtain a YKYY017 solution.

[0147] Example 3: Stabilizers (buffer solutions) with different pH values

[0148] On the basis of Example 1, in order to explore the influence of the pH value of the stabilizer (buffer solution) on the dry powder inhaler of YKYY017, change the pH value of the acetic acid-sodium acetate buffer solution in Step ①, and maintain the pH value of the solution obtained in Step ③ consistent with that of the buffer solution, and keep the prescription dosage unchanged. Change the pH value of the acetic acid-sodium acetate buffer solution in Step ① of Example 1, which is 3.7, to:

[0149] Method 1: The pH value of the acetic acid-sodium acetate buffer solution in Step ① is 3.0. After YKYY017 is dissolved in Step ③, add acetic acid solution or sodium hydroxide solution to adjust the pH value to 3.0;

[0150] Method 2: The pH value of the acetic acid-sodium acetate buffer solution in Step ① is 4.0. After YKYY017 is dissolved in Step ③, add acetic acid solution or sodium hydroxide solution to adjust the pH value to 4.0;

[0151] Method 3: The pH value of the acetic acid-sodium acetate buffer solution in Step ① is 2.5. After YKYY017 is dissolved in Step ③, add acetic acid solution or sodium hydroxide solution to adjust the pH value to 2.5;

[0152] Method 4: The pH value of the acetic acid-sodium acetate buffer solution in Step ① is 4.5. After YKYY017 is dissolved in Step ③, add acetic acid solution or sodium hydroxide solution to adjust the pH value to 4.5.

[0153] All other prescription ingredients and steps are the same as those in Example 1. The preparation methods of acetic acid-sodium acetate buffer solutions with different pH values and other components are as follows:

[0154] Table 4: Component table of Method 1 in Example 3

[0155]

[0156] Table 5: Component table of Method 2 in Example 3

[0157]

[0158] Table 6: Component table of Method 3 in Example 3

[0159]

[0160]

[0161] Table 7: Component table of Method 4 in Example 3

[0162]

[0163] Example 4: Selection of excipient types

[0164] On the basis of Example 1, in order to explore the influence of the type of excipient on the YKYY017 dry powder inhaler, change the type of excipient in the prescription while keeping the prescription dosage unchanged. Replace the excipient mannitol in Example 1 with:

[0165] Method 1: Trehalose;

[0166] Method 2: Sucrose;

[0167] The prescription dosage, other components of the prescription and the preparation method are the same as those in Example 1.

[0168] Example 5: Selection of carrier types

[0169] On the basis of Example 1, in order to explore the influence of the type of carrier on the YKYY017 dry powder inhaler, change the type of carrier in the prescription while keeping the prescription dosage unchanged. Replace the carrier lactose in Example 1 (D50: 106 μm; carrier source: DFE, Germany) with:

[0170] Method 1: Mannitol Pearlitol 100SD (D50: 80 - 120 μm), carrier source: Merck, Germany;

[0171] Method 2: Erythritol (D50: 80 - 120 μm), carrier source: Hunan Jiudian Hongyang;

[0172] Method 3: Xylitol (D50: 80 - 120 μm), carrier source: Hunan Jiudian Hongyang;

[0173] Method 4: Gum arabic spray-dried powder (D50: 80 - 120 μm), carrier source: NEXIRA, France.

[0174] The prescription dosage, other components of the prescription and the preparation method are the same as those in Example 1.

[0175] Example 6: Selection of carrier dosage

[0176] On the basis of Example 1, in order to explore the influence of the type of carrier on the YKYY017 dry powder inhaler, change the dosage of the carrier in the prescription. Replace the dosage of 31 parts of the carrier lactose in Example 1 with:

[0177] Method 1: 20 parts;

[0178] Method 2: 50 parts;

[0179] The other components of the prescription and the preparation method are the same as those in Example 1.

[0180] Example 7: Adding an anticoagulant

[0181] On the basis of Example 1, leucine, an anticoagulant, was added to explore the effect of the anticoagulant on the YKYY017 dry powder inhaler.

[0182] Prescription:

[0183] Table 8: Component table of Example 7

[0184]

[0185] Preparation method:

[0186] Step ①: Prepare an acetic acid-sodium acetate buffer solution with a pH of 3.7: Add sodium acetate and acetic acid to injection water and stir to dissolve to obtain the acetic acid-sodium acetate buffer solution;

[0187] Step ②: Dissolve mannitol in the above acetic acid-sodium acetate buffer solution with a pH of 3.7 and stir evenly to obtain a mannitol solution;

[0188] Step ③: Add YKYY017 to the above mannitol solution, stir to dissolve, and adjust the pH value to 3.7 to obtain a YKYY017 solution;

[0189] Step ④: Freeze-dry the above YKYY017 solution, control the water content < 1.0%, to obtain YKYY017 freeze-dried powder;

[0190] Step ⑤: Crush the above YKYY017 freeze-dried powder and leucine using a fluid energy mill, control the powder particle size, D90 is 1.0 - 5.0 μm, to obtain YKYY017 freeze-dried powder;

[0191] Step ⑥: Add the above YKYY017 freeze-dried powder and lactose to a high-speed stirring mixer, start stirring at 120 rpm, cutting at 2000 rpm, and mix for 10 min. After mixing, obtain the YKYY017 dry powder inhaler;

[0192] Example 8: Selection of anticoagulant types

[0193] In order to explore the effect of the type of anticoagulant on the inhalation preparation, when choosing different types of anticoagulants, on the basis of keeping the addition amount of the anticoagulant unchanged, the type of anticoagulant in Example 7 was replaced from leucine with:

[0194] Method 1: Mannitol;

[0195] Method 2: Poloxamer;

[0196] Method 3: Phospholipid;

[0197] The dosage in the prescription, other components of the prescription and the preparation method are the same as those in Example 7.

[0198] Example 9: Selection of the dosage of anticoagulant

[0199] To explore the influence of the dosage of anticoagulant on the inhalation preparation, when different dosages of anticoagulant were selected, the dosage of the anticoagulant leucine in Example 7 was replaced from 9 parts with:

[0200] Method 1: 5 parts;

[0201] Method 2: 18 parts;

[0202] Other components of the prescription and the preparation method are the same as those in Example 7.

[0203] Control Example 1:

[0204] Patent CN102264365A provides an inhalation preparation and its preparation method. Referring to Example 19 therein, the YKYY017 dry powder inhaler was prepared. The specific content is as follows:

[0205] Prescription:

[0206] Table 9: Component table of Control Example 1

[0207] Ingredient Function Weight Remarks YKYY017 Drug active ingredient 10 parts - Lactose Carrier 31 parts - Leucine Anticoagulant 9 parts -

[0208] Control Example 2:

[0209] Patent CN114727969A provides an inhalation preparation and its preparation method. Referring to Example 7 therein, the YKYY017 dry powder inhaler was prepared. The specific content is as follows:

[0210] Prescription:

[0211] Table 10: Component table of Control Example 2

[0212] Ingredient Function Weight Remarks YKYY017 Drug active ingredient 10 parts - Mannitol Excipient 20 parts - Water for injection Solvent 200 parts Water removed after lyophilization

[0213] Experimental Example 1: Investigation of fluidity

[0214] The fluidity of the powder sample was characterized by the angle of repose, bulk density and particle size.

[0215] The angle of repose (angle of repose) is the smallest angle formed with the horizontal surface when the inclined plane makes the object placed on it in a critical state of sliding along the inclined plane. That is, as the inclination angle increases, the object on the inclined plane will be more likely to slide; when the object reaches the state of starting to slide, the angle of this critical state is called the angle of repose. The smaller the angle of repose, the better the fluidity of the material.

[0216] The bulk density of the powder refers to: the mass of the powder per unit volume when the powder sample naturally fills a specified container.

[0217] Particle size D90: The particle size corresponding to when the cumulative particle size distribution of the sample reaches 90%. Its physical meaning is that particles with a particle size smaller than (or larger than) it account for 90%.

[0218] The smaller the bulk density, that is, the smaller or more irregular the particle size, the worse the material fluidity.

[0219] 1-1. Determination step ⑤ Obtaining the flowability data of YKYY017 lyophilized powder

[0220] Table 11: Flowability data of YKYY017 lyophilized powder

[0221] Example Angle of repose (°) <![CDATA[Bulk density (g / cm 3 )]]> Particle size D90 (μm) Example 1 50.2 0.17 2.93 Method 1 of Example 4 55.4 0.15 2.52 Method 2 of Example 4 58.1 0.13 3.12 Example 7 50.4 0.25 2.15 Method 1 of Example 9 57.1 0.19 3.16 Method 2 of Example 9 48.6 0.32 3.57

[0222] 1-2. Determination step ⑥ to obtain the flowability data of YKYY017 dry powder inhaler (preparation)

[0223] Table 12: Flowability data of YKYY017 dry powder inhaler

[0224] Example Angle of repose (°) Example 1 40.2 Method 1 of Example 5 43.1 Method 1 of Example 6 45.5 Method 2 of Example 6 36.4 Example 7 38.2 Method 1 of Example 8 44.3 Method 2 of Example 8 46.8 Method 3 of Example 8 39.9 Method 1 of Example 9 44.5 Method 2 of Example 9 38.6 Control Example 1 58.6 Control Example 2 61.3

[0225] Experimental Example 2: Hygroscopicity Study

[0226] Increased humidity can affect particle size and crystal morphology, causing powder aggregation and destroying its lung deposition and in vitro distribution characteristics, further affecting its stability and bioavailability.

[0227] The dry powder inhaler obtained in step ⑥ of different embodiments was placed in a stability test box at 25°C ± 2°C and 60% ± 5% RH for 24 hours, and its moisture absorption rate was calculated.

[0228] Table 13: Hygroscopicity data

[0229] Example Moisture absorption rate (%) Example 1 0.3 Method 1 of Example 5 0.5 Method 2 of Example 5 1.2 Method 3 of Example 5 2.5 Method 4 of Example 5 3.8

[0230] Experimental Example 3: In vitro lung deposition test (inhalation characteristics)

[0231] As a dry powder inhaler for pulmonary drug delivery, it needs to have certain inhalation characteristics to ensure that during the drug delivery process, the drug is dispersed with the carrier or other components by the airflow generated by the patient's inhalation, and the dispersed drug is delivered to the lungs to produce therapeutic effects. Generally, particles larger than 5 μm are likely to collide and settle in the throat, while particles smaller than 5 μm are more likely to be delivered to the bronchi. The particle size is a key factor affecting the inhalation performance. Therefore, it is necessary to study the ease of separation of the drug from other components in the dry powder inhaler and the drug particle size. According to General Chapter 0951 of the Fourth Part of the Chinese Pharmacopoeia (2020 Edition), in vitro, the aerodynamic characteristics of fine particles of inhalation preparations are used to determine the deposition state of the drug in the lungs, and the fine particle dose of the drug and the aerodynamic mass median diameter (D50) are used as evaluation indicators.

[0232] According to the method for determining the aerodynamic characteristics of fine particles of inhalation preparations in General Chapter 0951 of the Fourth Part of the Chinese Pharmacopoeia (2020 Edition), the fine particle dose is determined. Using the Andersen Cascade Impactor (ACI), the drug distribution and deposition amount during the drug delivery process are detected at an air flow rate of 30 L / min (the same as the time in the method for investigating the delivery dose uniformity, which is 8 s), and the fine particle dose (less than 4.7 μm) of the drug and the aerodynamic mass median diameter are calculated. The test method is as follows:

[0233] ① Connect the equipment, turn on the vacuum pump connected to the impactor ACI, and close the two-way solenoid valve.

[0234] ② Take a dose of dry powder inhaler (labeled amount is 10 mg) and place it in the mouthpiece adapter.

[0235] ③ Open the two-way solenoid valve for 8 s to aspirate the powder into the impactor.

[0236] ④ Wait for 8 s after inhalation and then turn off the vacuum pump.

[0237] ⑤ Extract the drug in the mouthpiece adapter, pre-separator, and L-shaped connecting tube with water respectively.

[0238] ⑥ Extract the drug on the inner wall of each level and the corresponding collection plate with water.

[0239] ⑦ Determine the amount of drug in the above solutions.

[0240] Based on the amount of drug in the above solutions, plot the cumulative deposition rate vs. particle size curve.

[0241] From the above curve, it can be obtained that:

[0242] I) Aerodynamic mass median diameter (D50): When the total mass of various particle sizes of particulate matter with an aerodynamic diameter smaller than a certain value accounts for 50% of the total mass of all particulate matter (i.e., the sum of the masses of all different particle sizes), this diameter is called the mass median diameter. That is, half of the particulate matter with this median diameter has a particle size smaller than this diameter, and half is larger than this diameter.

[0243] And calculate

[0244] II) Fine particle dose, that is, the mass percentage of the drug with a particle size below 4.7 μm in the total amount.

[0245] The calculated data are as follows:

[0246] Table 14: Aerodynamic characteristics of fine particles

[0247]

[0248]

[0249] Experimental Example 4: Investigation of delivery effect

[0250] The delivered dose is the amount of drug directly measured for a single unit dose, which is an important indicator for evaluating the difference between the actual delivered dose of the product and the labeled amount, and is an important way to reflect the amount of drug that patients may inhale. The delivered dose uniformity reflects the difference in the delivered dose of different inhalation devices among the same batch. By controlling the delivered dose and the delivered dose uniformity, the accuracy of the delivered dose during the use of the drug is ensured, and it is avoided that the single dose is too high, exceeding the safe dose, or the single dose is too low, not reaching the effective dose and failing to achieve the therapeutic effect.

[0251] According to the method for determining the delivered dose uniformity in General Chapter 0111 Inhalation Preparations of the Fourth Part of the Chinese Pharmacopoeia (2020 Edition). The test method is as follows:

[0252] ① Insert the inhalation device (capsule-type Accuhaler) into the adapter of the test device.

[0253] ② Adjust the air flow rate to 30 L / min.

[0254] ③ Insert a dose of dry powder inhaler (labeled amount 10 mg) into the adapter, press it once, turn on the vacuum pump, and aspirate for 8 seconds.

[0255] ④ Extract the drug in the filter paper, adapter, and sample collection tube with water.

[0256] ⑤ Repeat the determination 10 times.

[0257] ⑥ Determine the content of the drug in the above extract.

[0258] The average value of the 10 measurement results is obtained from the above results, and the following are calculated:

[0259] I) The percentage of the mean value to the labeled amount, that is, the delivered dose.

[0260] (The defined standard is 80% - 120%; the closer the delivered dose is to 100%, the closer the actual delivered dose is to the labeled amount, and the more accurate the delivered dose is.)

[0261] And

[0262] II) Among different measurement results, the percentage of the measured value to the mean value is calculated, and its distribution range is statistically analyzed, that is, the delivered dose uniformity.

[0263] (The defined standard is 75% - 125%; the narrower its distribution range, the higher the delivered dose uniformity.)

[0264] The calculation results are as follows:

[0265] Table 15: Delivery effect data

[0266]

[0267]

[0268] Experimental Example 5: Stability study

[0269] The dry powder inhalants prepared in the examples and the control example are placed in a stability test chamber at 25°C ± 2°C and 75% ± 5% RH for 6 months. The contents of the drug active ingredient (YKYY017) and the fine particle dose before and after placement are measured respectively, and the change rate data before and after placement are calculated for comparison;

[0270] Among them, the change rate = |accelerated for 6 months - 0 days|. The results are as follows:

[0271] Table 16: Stability data - content of drug active ingredient

[0272]

[0273] Table 17: Stability data - fine particle dose

[0274]

[0275] Conclusion analysis 1: (Type of stabilizer)

[0276] In Example 1, YKYY017 freeze-dried powder was prepared using an acetic acid - acetate buffer system with a pH of 3.7, and then a dry powder inhalant was prepared. On this basis, the effects of different buffer systems on the resulting preparation were investigated. The inhalation characteristics, delivery effect, and stability of the resulting dry powder inhalant (preparation) are as follows:

[0277] 1-1. Differences in formulation prescriptions

[0278] Table 18: Differences in formulation prescriptions

[0279] Example Type of stabilizer (buffer system) Example 1 pH 3.7 acetic acid - acetate buffer solution Method 1 of Example 2 pH 3.7 citric acid - citrate buffer solution Method 2 of Example 2 pH 3.7 hydrochloric acid solution

[0280] 1-2. Inhalation characteristics

[0281] Table 19: Inhalation characteristics

[0282] Example Aerodynamic mass median diameter (μm) Fine particle dose (%) Limit standard ≤5 ≥10 Example 1 3.04 30.1 Method 1 of Example 2 3.59 25.6 Method 2 of Example 2 2.76 32.3

[0283] 1-3. Delivery effect

[0284] Table 20: Delivery effect

[0285]

[0286] 1-4. Stability

[0287] Table 21: Stability

[0288]

[0289] Conclusion:

[0290] 1) The results of the inhalation characteristics and delivery effect of the formulation show that:

[0291] In Example 1, a pH 3.7 acetic acid - acetate buffer solution was used as a stabilizer, and the resulting formulation had a small aerodynamic mass median diameter of the drug, which was 3.04 μm, far less than the limit standard (≤5 μm); the proportion of the fine particle dose of the drug was large, which was 30.1%, far higher than the limit standard (≥10%); the above results indicate that the prepared dry powder inhaler can be fully delivered to the bronchi to exert its medicinal effect and has good pulmonary deposition characteristics.

[0292] The delivery dose was relatively high, with an average value accounting for 89% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the average value was distributed between 80% and 116% (required to be between 75% and 125%), and the distribution was concentrated, indicating good uniformity of the delivery dose.

[0293] In Example 2, Method 1 used a pH 3.7 citric acid - citrate buffer solution as a stabilizer, and the resulting formulation had good inhalation characteristics. The aerodynamic mass median diameter of the drug in the resulting formulation was small, which was 3.59 μm, far less than the limit standard (≤5 μm); the fine particle dose of the drug was relatively large, which was 25.6%, far higher than the limit standard (≥10%).

[0294] The delivered dose is high, with the mean accounting for 88% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the mean is distributed between 77% and 117% (required to be between 75% and 125%), the distribution is concentrated, and the uniformity of the delivered dose is good.

[0295] In Example 2, Method 2 uses a pH 3.7 hydrochloric acid solution as a stabilizer. The aerodynamic mass median diameter of the drug in the obtained preparation is small, being 2.76 μm, which is less than the limit standard (≤5 μm); the dose of fine drug particles is large, being 32.3%, which is higher than the limit standard (≥10%).

[0296] The delivered dose is relatively high, with the mean accounting for 84% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the mean is distributed between 88% and 123% (required to be between 75% and 125%), the distribution is concentrated, and the uniformity of the delivered dose is good.

[0297] 2) The stability results show that:

[0298] In Example 1, a pH 3.7 acetic acid - acetate buffer solution is used as a stabilizer and placed at the accelerated test conditions (40°C ± 2°C, 75% ± 5% RH) for 6 months. Comparing day 0 with 6 months of acceleration, the contents of the drug active substance are 98.4% and 97.9% respectively, and the change rate is 0.5% (<5%), without obvious changes, indicating that the obtained preparation has good stability.

[0299] In Example 2, Method 1 uses a pH 3.7 citric acid - citrate buffer solution as a stabilizer and placed at the accelerated test conditions (40°C ± 2°C, 75% ± 5% RH) for 6 months. Comparing day 0 with 6 months of acceleration, the contents are 99.2% and 99.3% respectively, and the change rate is 0.1% (<5%), being basically the same, indicating that the obtained preparation has good stability.

[0300] In Example 2, Method 2 uses a pH 3.7 hydrochloric acid solution as a stabilizer and placed at the accelerated test conditions (40°C ± 2°C, 75% ± 5% RH) for 6 months. Comparing day 0 with 6 months of acceleration, the contents are 99.6% and 85.4% respectively, and the change rate is 14.2% (>5%), with significant changes, indicating that the obtained preparation has poor stability and cannot be used as a qualified reagent.

[0301] From the comparison of the inhalation characteristics and stability results of the above preparations, it can be seen that although the inhalation characteristics prepared using the above stabilizers all meet the limit standards; however, when a pH 3.7 hydrochloric acid solution is used as a stabilizer in Example 2, Method 2, the obtained preparation has poor stability and cannot prepare qualified preparation products.

[0302] Summary:

[0303] 1. A dry powder inhaler is prepared using a buffer system as a stabilizer (for example, the acetic acid - acetate solution in Example 1, and the citric acid - citrate buffer solution in Method 1 of Example 2). The resulting preparation has good inhalation characteristics and delivery effects of the drug, and remarkable stability, specifically manifested as follows:

[0304] i. The median aerodynamic mass diameter of the drug in the resulting preparation is small, only 3.04 μm to 3.59 μm, less than 5 μm (particles that can enter the lungs are below 5 μm); and the dose of fine drug particles is large, 25.6% to 30.1%, far higher than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles to enter the lungs.

[0305] ii. The resulting preparation has a high delivery dose and good delivery dose uniformity. The delivery dose is 88% to 89%; the delivery dose uniformity is distributed between 77% and 117%, with a narrow distribution range, higher than the limit standard (75% - 125%).

[0306] iii. The resulting preparation has good stability. When placed under accelerated test conditions (40°C ± 2°C, 75% ± 5% RH) for 6 months, compared with the data on day 0, the content change rate is low, only 0.1% to 0.5%, far less than 5%.

[0307] 2. A dry powder inhaler is prepared using other solutions or solutions without buffering capacity as stabilizers (for example, the hydrochloric acid solution in Method 2 of Example 2). The drug has poor stability and cannot be used as a qualified preparation product. Specifically manifested as follows:

[0308] i. The resulting preparation has poor stability. When placed under accelerated test conditions (40°C ± 2°C, 75% ± 5% RH) for 6 months, compared with the data on day 0, the content change rate is significant, 14.2%, far greater than the limit standard (≤5%), and it cannot be used as a qualified dry powder inhaler.

[0309] Conclusion Analysis 2: (pH of the stabilizer)

[0310] As can be seen from the results in Conclusion Analysis 1, in Example 1, a pH 3.7 acetic acid - acetate buffer solution is used as the buffer system, and the resulting dry powder inhaler has good inhalation characteristics and stability. Therefore, on the basis of Example 1, the pH value of the buffer system is changed to explore its effects on the inhalation characteristics, delivery effects, and stability of the dry powder inhaler. The results are as follows:

[0311] 2 - 1. Differences in the formulation of the preparation

[0312] Table 22: Differences in the formulation of the preparation

[0313]

[0314]

[0315] 2-2. Inhalation characteristics of the preparation

[0316] Table 23: Inhalation characteristics of the preparation

[0317] Example Aerodynamic mass median diameter (μm) Fine particle dose (%) Example 1 3.04 30.1 Method 1 of Example 3 2.69 27.2 Method 2 of Example 3 3.26 25.5 Method 3 of Example 3 3.61 29.6 Method 4 of Example 3 4.20 24.9

[0318] 2-3. Delivery effect

[0319] Table 24: Delivery effect

[0320]

[0321] 2-4. Stability

[0322] Table 25: Stability

[0323]

[0324]

[0325] Conclusion:

[0326] 1) The results of the inhalation characteristics and delivery effect of the preparation show that:

[0327] In Example 1, a pH 3.7 acetic acid - acetate buffer solution was used as a stabilizer. The resulting preparation had a small aerodynamic mass median diameter of the drug, which was 3.04 μm, far less than the limit standard (≤5 μm); the proportion of the fine particle dose of the drug was large, which was 30.1%, far higher than the limit standard (≥10%); the above results indicate that the prepared dry powder inhaler can be fully delivered to the bronchi to exert its medicinal effect and has good lung deposition characteristics.

[0328] The delivery dose was relatively high, with an average value accounting for 89% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the average value was distributed between 80% and 116% (required to be between 75% and 125%), and the distribution was concentrated, indicating good uniformity of the delivery dose.

[0329] In Example 3 Method 1 and Example 3 Method 2, pH 3.0 and pH 4.0 acetic acid - acetate buffer solutions were used as stabilizers respectively. The resulting preparations had small aerodynamic mass median diameters of the drug, which were 2.69 μm and 3.26 μm respectively, far less than the limit standard (≤5 μm); the proportion of the fine particle dose of the drug was large, controlled at about 25 - 27%, far higher than the limit standard (≥10%); the above results indicate that the prepared dry powder inhaler can be fully delivered to the bronchi to exert its medicinal effect and has good lung deposition characteristics.

[0330] The delivered dose is relatively high, with the mean accounting for 85% - 89% of the labeled amount (required to be between 80% - 120%); the percentage range of the measured value to the mean is distributed between 79% - 119% (required to be between 75% - 125%), the distribution is concentrated, and the uniformity of the delivered dose is good.

[0331] For Method 3 and Method 4 of Example 3, acetic acid - acetate buffer solutions with pH 2.5 and pH 4.5 were used as stabilizers respectively. The aerodynamic mass median diameter of the drug in the obtained preparations showed an increasing trend compared with Example 1, being 3.61 μm - 4.20 μm; the proportion of the fine particle dose of the drug was relatively large, about 24% - 29%, meeting the limit standard (≥10%);

[0332] Among the delivered doses, the mean accounts for about 80% - 90% of the labeled amount (required to be between 80% - 120%); the percentage range of the measured value to the mean is distributed between 76% - 124% (required to be between 75% - 125%). The distribution range has slightly expanded, but still meets the uniformity of the delivered dose.

[0333] 2) The stability results show that:

[0334] In Example 1, acetic acid - acetate buffer solution with pH 3.7 was used as a stabilizer and placed at accelerated test conditions (40°C ± 2°C, 75% ± 5% RH) for 6 months. Comparing the drug active substance content on day 0 and after 6 months of acceleration, they were 98.4% and 97.9% respectively, with a change rate of 0.5% (<5%), indicating no obvious change, which shows that the obtained preparation has good stability.

[0335] For the dry powder inhaler prepared by Method 1 of Example 3 using acetic acid - acetate buffer solution with pH 3.0 as a stabilizer and placed at accelerated test conditions (40°C ± 2°C, 75% ± 5% RH) for 6 months, comparing the content on day 0 and after 6 months of acceleration, they were 99.2% and 97.3% respectively, with a change rate of 1.9%, less than 5%, indicating no obvious change, further showing that the obtained preparation has good and significant stability.

[0336] For the dry powder inhaler prepared by Method 2 of Example 3 using acetic acid - acetate buffer solution with pH 4.0 as a stabilizer and placed at accelerated test conditions (40°C ± 2°C, 75% ± 5% RH) for 6 months, comparing the content on day 0 and after 6 months of acceleration, they were 98.6% and 96.9% respectively, with a change rate of 1.7%, less than 5%, indicating no obvious change, further showing that the obtained preparation has good and significant stability.

[0337] Example 3 Method 3 used a pH 2.5 acetic acid - acetate buffer solution and Example 3 Method 4 used a pH 4.5 acetic acid - acetate buffer solution as stabilizers to prepare dry powder inhalants. They were placed at accelerated test conditions (40°C ± 2°C, 75% ± 5% RH) for 6 months. When comparing day 0 with 6 months of acceleration, the contents of Example 3 Method 3 were 97.9% and 82.3% respectively, with a change rate of 15.6%, showing a significant change, far higher than the limit standard (≤5%); the contents of Example 3 Method 4 were 99.4% and 89.4% respectively, with a change rate of 10.0%, showing a significant change (much greater than 5%); indicating that the obtained preparations had poor stability and could not be used as qualified reagents.

[0338] Summary:

[0339] 1. Using a buffer system with a pH of 3.0 - 4.0 (for example, Example 1 pH 3.7, Example 3 Method 1 pH 3.0, Method 2 pH 4.0) as a stabilizer to prepare dry powder inhalants, the obtained preparations had good inhalation characteristics and delivery effects, and good and significant stability, specifically manifested as:

[0340] i. The drug aerodynamic mass median diameter of the obtained preparations was small, ranging from 2.69 μm to 3.26 μm, less than 5 μm; the fine particle dose of the drug in the obtained preparations was large, ranging from 25.5% to 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there were sufficient drug particles to enter the lungs;

[0341] ii. The obtained preparations had a high delivery dose and good delivery dose uniformity. The delivery dose was 85% - 89%; the delivery dose uniformity was distributed in the range of 79% - 119%, with a narrow distribution range, higher than the limit standard (75% - 125%);

[0342] iii. The obtained preparations had good stability. After the accelerated test, the change rate of the drug active ingredient content was small, ranging from 0.5% to 1.9%, less than 5%.

[0343] 2. When using a buffer system with a pH of 3.7 (for example, Example 1 pH 3.7) as a stabilizer, the obtained preparations had the best inhalation characteristics and delivery effects, and the optimal stability; specifically manifested as:

[0344] i. The drug aerodynamic mass median diameter of the obtained preparations was small, being 3.04 μm, less than 5 μm; the fine particle dose of the drug in the obtained preparations was large, being 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there were sufficient drug particles to enter the lungs;

[0345] ii. The obtained preparations had a high delivery dose and good delivery dose uniformity. The delivery dose was 89%; the delivery dose uniformity was distributed between 80% - 116%, with a narrow distribution range, far higher than the limit standard of 75% - 125%;

[0346] iii. The stability of the obtained preparation is optimal. After the accelerated test, the change rate of the content of the drug active ingredient is small, which is 0.5%, far less than 5%, and there is basically no change.

[0347] 3. When the pH value of the buffer system is less than 3.0 (for example, pH 2.5 in Method 3 of Example 3) or when the pH value of the buffer system is greater than 4.0 (for example, pH 4.5 in Method 4 of Example 3), the stability of the obtained preparation is poor and it cannot be used as a qualified preparation product. The specific manifestations are as follows:

[0348] i. The stability of the obtained preparation is poor. After the accelerated test, the change rate of the content of the drug active ingredient is large, which is 10.0% - 15.6%, far greater than 5%, and significant changes have occurred, so it cannot be used as a qualified dry powder inhaler.

[0349] Conclusion Analysis 3: (Types of excipients)

[0350] In Example 1, mannitol was used as the excipient, and the obtained dry powder inhaler had good inhalation characteristics, delivery effect and stability. Therefore, on the basis of Example 1, the types of excipients were changed to explore their effects on the fluidity of the freeze-dried powder and the inhalation characteristics and delivery effect of the dry powder inhaler. The results are as follows:

[0351] 3-1. Differences in the preparation prescriptions

[0352] Table 26: Differences in the preparation prescriptions

[0353] Example Excipient Example 1 Mannitol Method 1 of Example 4 Trehalose Method 2 of Example 4 Sucrose

[0354] 3-2. Fluidity of the freeze-dried powder

[0355] Table 27: Fluidity of the freeze-dried powder

[0356]

[0357]

[0358] 3-3. Inhalation characteristics of the dry powder inhaler

[0359] Table 28: Inhalation characteristics of the dry powder inhaler

[0360] Example Aerodynamic mass median diameter (μm) Fine particle dose (%) Example 1 3.04 30.1 Method 1 of Example 4 4.49 23.2 Method 2 of Example 4 4.96 19.5

[0361] 3-4. Delivery effect of the dry powder inhaler

[0362] Table 29: Delivery effect of the dry powder inhaler

[0363]

[0364] Conclusion:

[0365] 1) The fluidity results of the YKYY017 freeze-dried powder show that:

[0366] In Example 1, mannitol was used as an excipient, and the obtained YKYY017 freeze-dried powder had a repose angle of 50.2°, a bulk density of 0.17 g / cm 3 , a particle size D90 of 2.93 μm, small particles and a small repose angle. Therefore, the freeze-dried powder has good fluidity.

[0367] In Example 4, methods 1 and 2 used trehalose and sucrose as excipients, and the obtained YKYY017 freeze-dried powder had a repose angle of about 55-58°, a bulk density of 0.13 g / cm 3 ~0.15 g / cm 3 , a particle size D90 of 2.52 μm - 3.12 μm, small particles and a small repose angle, and also had good fluidity.

[0368] 2) The inhalation characteristics and delivery effects of the dry powder inhaler show that:

[0369] In Example 1, mannitol was used as an excipient. The median aerodynamic mass diameter of the drug in the obtained preparation was small, 3.04 μm, much smaller than the limit standard (≤5 μm); the proportion of the fine particle dose of the drug was large, 30.1%, much higher than the limit standard (≥10%); the above results indicate that the prepared dry powder inhaler can be fully delivered to the bronchus to exert its medicinal effect and has good pulmonary deposition characteristics.

[0370] The delivered dose was relatively high, with an average value accounting for 89% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the average value was distributed between 80% and 116% (required to be between 75% and 125%), with a concentrated distribution and good uniformity of the delivered dose.

[0371] In Example 4, methods 1 and 2 used trehalose and sucrose as excipients respectively. The median aerodynamic mass diameter of the drug in the obtained preparations was also small, 4.49 μm and 4.96 μm, within the limit standard (≤5 μm); the proportion of the fine particle dose of the drug was large, 23.2% and 19.5% respectively, higher than the limit standard (≥10%); the above results indicate that the prepared dry powder inhaler can be fully delivered to the bronchus to exert its medicinal effect and has good pulmonary deposition characteristics.

[0372] The delivered dose was relatively high, with an average value accounting for 84% - 90% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the average value was distributed between 75% and 125%, with a concentrated distribution and good uniformity of the delivered dose.

[0373] Summary:

[0374] 1. The dry powder inhalants prepared with the excipients of the present invention (for example, mannitol in Example 1, trehalose in Method 1 of Example 4, and sucrose in Method 2 of Example 4) have good fluidity of the obtained freeze-dried powder, and excellent inhalation characteristics and delivery effects. Specifically, it is manifested as follows:

[0375] i. It has good fluidity, a small angle of repose, which is 50.2° - 58.1°; the bulk density is 0.13 g / cm 3 - 0.17 g / cm 3 ; small particles, the particle size D90 is 2.52 μm - 3.12 μm; it has good fluidity;

[0376] ii. The median aerodynamic mass diameter of the drug in the obtained preparation is small, which is 3.04 μm - 4.96 μm, less than 5 μm; the fine particle dose of the drug in the obtained preparation is large, which is 19.5% - 30.1%, greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles to enter the lungs;

[0377] iii. The obtained preparation has a high delivery dose and good delivery dose uniformity, the delivery dose is 84% - 90%; the delivery dose uniformity is distributed between 75% - 125%.

[0378] 2. Using mannitol (for example, in Example 1) as the excipient, the obtained freeze-dried powder has excellent fluidity, and the inhalation characteristics and delivery effects of the preparation are the best. Specifically, it is manifested as follows:

[0379] i. It has the best fluidity, and the angle of repose is as low as 50.2°; the bulk density is 0.17 g / cm 3 ; small particles, the particle size D90 is 2.93 μm;

[0380] ii. The median aerodynamic mass diameter of the drug in the obtained preparation is small, which is 3.04 μm, less than 5 μm; the fine particle dose of the drug in the obtained preparation is large, which is 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles to enter the lungs;

[0381] iii. The obtained preparation has a high delivery dose and good delivery dose uniformity, the delivery dose is 89%; the delivery dose uniformity is distributed between 80% - 116%, the distribution range is narrow, far higher than the limit standard of 75% - 125%.

[0382] Conclusion Analysis 4: (Carrier type)

[0383] As can be seen from the results in Conclusion Analyses 1-3, lactose was used as the carrier in Example 1, and the resulting dry powder inhaler had good inhalation characteristics and delivery effects. Therefore, on the basis of Example 1, the type of carrier was changed to explore its effects on the hygroscopicity, inhalation characteristics, delivery effects, and stability of the dry powder inhaler. The results are as follows:

[0384] 4-1. Differences in formulation prescriptions

[0385] Table 30: Differences in formulation prescriptions

[0386] Example Carrier type Example 1 31 parts of lactose (D50: 106 μm) Method 1 of Example 5 31 parts of mannitol Pearlitol 100SD (D50: 113 μm) Method 2 of Example 5 31 parts of erythritol (D50: 99 μm) Method 3 of Example 5 31 parts of xylitol (D50: 110 μm) Method 4 of Example 5 31 parts of arabic gum spray-dried powder (D50: 92 μm)

[0387] 4-2. Hygroscopicity of the dry powder inhaler (Step ⑥)

[0388] Table 31: Hygroscopicity data

[0389] Example Moisture absorption rate (%) Example 1 0.3 Method 1 of Example 5 0.5 Method 2 of Example 5 1.2 Method 3 of Example 5 2.5 Method 4 of Example 5 3.8

[0390] 4-3. Inhalation characteristics of the dry powder inhaler

[0391] Table 32: Inhalation characteristic data of the formulation

[0392] Example Aerodynamic mass median diameter (μm) Fine particle dose (%) Example 1 3.04 30.1 Method 1 of Example 5 3.56 27.5 Method 2 of Example 5 4.52 21.6 Method 3 of Example 5 4.28 28.9 Method 4 of Example 5 4.63 22.3

[0393] 4-4. Delivery effects of the dry powder inhaler

[0394] Table 33: Delivery effects of the dry powder inhaler

[0395]

[0396] 4-5. Stability of the dry powder inhaler

[0397] Table 34: Stability of the dry powder inhaler

[0398]

[0399] Conclusion:

[0400] 1) The hygroscopicity results of the dry powder inhaler (Step ⑥) show that:

[0401] In Example 1 and Method 1 of Example 5, lactose and mannitol were used as carriers respectively. After the resulting dry powder inhalers were placed in a stability test chamber at 25°C ± 2°C and 60% ± 5% RH for 24 hours, the hygroscopic rates were 0.3% and 0.5% respectively, and the hygroscopicity was low, probably because the hygroscopicity of lactose and mannitol is small.

[0402] In Example 5, erythritol, xylitol, and arabic gum were used as carriers in Method 2, Method 3, and Method 4 respectively. After the obtained dry powder inhalants were placed in a stability test chamber at 25°C ± 2°C and 60% ± 5% RH for 24 hours, the moisture absorption rates were 1.2%, 2.5%, and 3.8% respectively. The hygroscopicity increased slightly compared with that of Example 1 and Method 1 of Example 8, probably because erythritol, xylitol, and arabic gum have a certain hygroscopicity, but still meet the standard (not exceeding 5%).

[0403] 2) Inhalation characteristics and delivery effects

[0404] In Example 1, mannitol was used as an excipient. The median aerodynamic mass diameter of the drug in the obtained preparation was small, 3.04 μm, much smaller than the limit standard (≤5 μm); the proportion of fine particle dose of the drug was large, 30.1%, much higher than the limit standard (≥10%); the above results indicate that the prepared dry powder inhalant can be fully delivered to the bronchus to exert its drug effect and has good pulmonary deposition characteristics.

[0405] The delivery dose was high, and the average value accounted for 89% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the average value was distributed between 80% and 116% (required to be between 75% and 125%), and the distribution was concentrated, and the delivery dose uniformity was good.

[0406] In Example 5, mannitol, erythritol, xylitol, and arabic gum were used as carriers in Method 1 to Method 4 respectively. The median aerodynamic mass diameter of the drug was 3.56 μm to 4.63 μm, less than the limit standard (≤5 μm);

[0407] The proportion of fine particle dose of the drug was controlled between 21.6% and 28.9%, higher than the limit standard (≥10%); the above results indicate that the prepared dry powder inhalant can be fully delivered to the bronchus to exert its drug effect and has good pulmonary deposition characteristics.

[0408] The delivery dose was relatively high, and the average value accounted for 82% to 90% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the average value was distributed between 75% and 124%, and the distribution was relatively concentrated, and the delivery dose uniformity was good.

[0409] 3) The stability results show that:

[0410] In Example 1 and Method 1 of Example 5, lactose and mannitol were used as carriers respectively. After the obtained preparations were placed under accelerated test conditions (25°C ± 2°C, 75% ± 5% RH) for 6 months, compared with day 0, the fine particle doses decreased from 30.1% to 28.3% and from 27.5% to 25.3% respectively, and the change rates were 1.8% and 2.2% respectively. The change rates were much less than 5%, and the stability was good.

[0411] In Example 5, for Method 2, Method 3, and Method 4, erythritol, xylitol, and arabic gum were used as carriers respectively. The prepared preparations were placed under accelerated test conditions (25°C ± 2°C, 75% ± 5% RH) for 6 months. Compared with day 0, the fine particle doses decreased from 21.6% to 19.2%, from 28.9% to 25.4%, and from 22.3% to 18.1% respectively, and the change rates were 2.4%, 3.5%, and 4.2% respectively. All the change rates were less than 5%, showing good stability effects.

[0412] Summary:

[0413] 1. The dry powder inhalants prepared using the carriers of the present invention (such as lactose, mannitol, erythritol, xylitol, and arabic gum in Example 1 and Method 1 - 4 of Example 5) have good hygroscopicity, excellent inhalation characteristics and delivery effects, and excellent stability. Specifically, it is manifested as follows:

[0414] i. The prepared preparations have good hygroscopicity, and the moisture absorption rate is controlled within 0.3% - 3.8%, less than 5%, indicating low hygroscopicity.

[0415] ii. The aerodynamic mass median diameter of the drug in the prepared preparations is small, ranging from 3.04 μm to 4.63 μm, less than 5 μm; the fine particle dose of the drug in the prepared preparations is large, ranging from 21.6% to 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles to enter the lungs.

[0416] iii. The prepared preparations have a high delivery dose and good delivery dose uniformity. The delivery dose is 82% - 90%; the delivery dose uniformity is distributed between 75% - 124%, with a narrow distribution range.

[0417] iv. The prepared preparations have good stability. After the accelerated test, the fine particle dose of the drug is 18.1% - 28.3%; the change rate is controlled within 1.8% - 4.2%, less than 5%.

[0418] 2. The dry powder inhalants prepared using lactose and mannitol as carriers have the lowest hygroscopicity, excellent inhalation characteristics and delivery effects, and the best stability. Specifically, it is manifested as follows:

[0419] i. The prepared preparations have good hygroscopicity, and the moisture absorption rate is controlled within 0.3% - 0.5%, significantly lower than 5%.

[0420] The hygroscopicity is optimal.

[0421] ii. The median aerodynamic mass diameter of the obtained preparation is small, ranging from 3.04 μm to 3.56 μm, less than 5 μm; the fine particle dose of the obtained preparation is large, ranging from 27.5% to 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles to enter the lungs.

[0422] iii. The obtained preparation has a high delivery dose and good delivery dose uniformity. The delivery dose is 83% - 89%; the delivery dose uniformity is distributed between 80% and 124%, with a narrow distribution range.

[0423] iv. The obtained preparation has the best stability. After the accelerated test, the fine particle dose of the drug is between 25.3% and 28.3%; the change rate is only 1.8% - 2.2%, far less than 5%.

[0424] Conclusion Analysis 5: (Carrier dosage)

[0425] As can be seen from the results in Conclusion Analysis 1, lactose was used as the carrier in Example 1, and the obtained dry powder inhaler had good inhalation characteristics and delivery effects. Therefore, on the basis of Example 1, the dosage of the carrier lactose was changed to explore its effects on the fluidity, inhalation characteristics, and delivery effects of the dry powder inhaler. The results are as follows:

[0426] 5-1. Differences in the formulation prescription (carrier dosage)

[0427] Table 35: Differences in the formulation prescription (carrier dosage)

[0428] Example Carrier dosage Example 1 31 parts Method 1 of Example 6 20 parts Method 2 of Example 6 50 parts

[0429] 5-2. Fluidity of the dry powder inhaler (step ⑥)

[0430] Table 36: Fluidity of the dry powder inhaler (step ⑥)

[0431] Example Angle of repose (°) Example 1 40.2 Method 1 of Example 6 45.5 Method 2 of Example 6 36.4

[0432] 5-3. Inhalation characteristics

[0433] Table 37: Inhalation characteristics

[0434] Example Aerodynamic mass median diameter (μm) Fine particle dose (%) Example 1 3.04 30.1 Method 1 of Example 6 3.36 26.5 Method 2 of Example 6 4.54 23.3

[0435] 5-4. Delivery effects

[0436] Table 38: Delivery effects

[0437]

[0438]

[0439] Conclusion:

[0440] 1) The results of the flowability of the dry powder inhaler show that:

[0441] In Example 1 and Example 6 (Method 1, Method 2), 31 parts, 20 parts, and 50 parts of lactose were used as carriers respectively, and there were differences in the angle of repose of the resulting preparations, and the differences were obvious. Among them, the angles of repose of the dry powder inhalers with 20 parts, 31 parts, and 50 parts of lactose were 45.5°, 40.2°, and 36.4° respectively. The small angle of repose indicates good flowability.

[0442] 2) The results of the aerodynamic mass median diameter of the drug show that:

[0443] When the carrier dosage in Example 1 was 31 parts, the aerodynamic mass median diameter of the resulting preparation was small, 3.04 μm, far less than the limit standard (≤5 μm); the proportion of the fine particle dose of the drug was large, 30.1%, far higher than the limit standard (≥10%); the above results indicate that the prepared dry powder inhaler can be fully delivered to the bronchus to exert its efficacy and has good pulmonary deposition characteristics.

[0444] In Example 6, Method 1 and Method 2 used 20 parts and 50 parts of lactose as carriers respectively, and the aerodynamic mass median diameters of the resulting preparations were 3.36 μm and 4.54 μm respectively, both less than 5 μm. The proportion of the fine particle dose of the drug was large, 23.3% - 26.5%, far higher than the limit standard (≥10%); the above results indicate that the prepared dry powder inhaler can be fully delivered to the bronchus to exert its efficacy and has good pulmonary deposition characteristics.

[0445] 3) The delivery effect shows that:

[0446] When the carrier dosage in Example 1 was 31 parts, the delivery dose of the resulting preparation was high, and the average value accounted for 89% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the average value was distributed between 80% and 116% (required to be between 75% and 125%), and the distribution was concentrated, indicating good uniformity of the delivery dose.

[0447] In Example 6, when Method 1 and Method 2 used 20 parts and 50 parts of lactose as carriers respectively, the delivery doses of the resulting preparations were high, and the average values accounted for 83% - 87% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the average value was distributed between 85% and 119% (required to be between 75% and 125%), and the distribution was concentrated, indicating good uniformity of the delivery dose.

[0448] Summary:

[0449] 1. The dry powder inhalant prepared by adding 20 - 50 parts of carrier (for example, 31 parts in Example 1, 20 parts in Method 1 of Example 6, and 50 parts in Method 2 of Example 6) has good powder fluidity, inhalation characteristics, and delivery effect. Specifically, it is shown in the following aspects:

[0450] i. The angle of repose of the obtained preparation is small, controlled within 36.4° - 45.5°, and the material fluidity is good.

[0451] ii. The median aerodynamic mass diameter of the drug in the obtained preparation is small, ranging from 3.04 μm to 4.54 μm, less than 5 μm; the fine particle dose of the drug in the obtained preparation is large, ranging from 23.3% to 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles to enter the lungs.

[0452] iii. The obtained preparation has a high delivery dose and good delivery dose uniformity. The delivery dose is 83% - 89%; the delivery dose uniformity is distributed between 80% - 119%, with a narrow distribution range, far higher than the limit standard of 75% - 125%.

[0453] 2. The dry powder inhalant prepared with 30 - 35 parts of carrier (for example, 31 parts in Example 1) has the best inhalation characteristics. Specifically, it is shown in the following aspects:

[0454] i. The angle of repose of the obtained preparation is relatively small, being 40.2°, and the material fluidity is good.

[0455] ii. The median aerodynamic mass diameter of the drug in the obtained preparation is small, being 3.04 μm, less than 5 μm; the fine particle dose of the drug in the obtained preparation is large, being 30.1%, far greater than the pharmacopoeia requirement (≥10%), indicating that there are sufficient drug particles to enter the lungs.

[0456] iii. The obtained preparation has a high delivery dose and good delivery dose uniformity. The delivery dose is 89%; the delivery dose uniformity is distributed between 80% - 116%, with a narrow distribution range, far higher than the limit standard of 75% - 125%.

[0457] Conclusion Analysis 6: (Whether to add anticoagulant)

[0458] In Example 1 and Example 7, dry powder inhalants are prepared without using anticoagulant and with 9 parts of anticoagulant respectively. The fluidity data of the freeze - dried powder in step ⑤, the fluidity, inhalation characteristics, delivery dose, and stability of the dry powder inhalant (preparation) are as follows:

[0459] 6 - 1. Differences in the formulation of the preparation

[0460] Table 39: Differences in the formulation of the preparation

[0461] Example Anticoagulant addition amount Example 1 No anticoagulant leucine Example 7 Leucine dosage is 9 parts

[0462] 6-2. Flowability of the freeze-dried powder (Step ⑤)

[0463] Table 40: Flowability of the freeze-dried powder (Step ⑤)

[0464]

[0465]

[0466] 6-3. Flowability of the dry powder inhaler (Step ⑥)

[0467] Table 41: Flowability of the dry powder inhaler (Step ⑥)

[0468] Example Angle of repose (°) Example 1 40.2 Example 7 38.2

[0469] 6-4. Inhalation characteristics of the dry powder inhaler

[0470] Table 42: Inhalation characteristics of the dry powder inhaler

[0471] Example Aerodynamic mass median diameter (μm) Fine particle dose (%) Example 1 3.04 30.1 Example 7 2.59 42.6

[0472] 6-5. Delivery effect of the dry powder inhaler

[0473] Table 43: Delivery effect of the dry powder inhaler

[0474]

[0475] 6-6. Stability of the dry powder inhaler

[0476] Table 44: Stability of the dry powder inhaler

[0477]

[0478] Conclusion:

[0479] 1) Flowability:

[0480] 1-1) The flowability results of the YKYY017 freeze-dried powder show that:

[0481] In Example 1, without adding anticoagulant, after the freeze-dried powder was directly pulverized by a pneumatic mill, the material texture was light, the static electricity between the powders was obvious, it was easy to agglomerate and cake, the loose bulk density was small, and the angle of repose was large (50.2°), which would lead to uneven mixing with the carrier or difficulty in separating from the carrier during use, thus affecting the total amount of drug entering the lungs.

[0482] In Example 7, leucine was added as an anticoagulant. After being pulverized with the fluid energy mill together with the freeze-dried powder, the texture of the material remained light. Among them, leucine played an antistatic role, significantly improving the state of material agglomeration and caking. The angle of repose was significantly lower than that in Example 1 without an anticoagulant, being 50.4°, and the angle of repose did not change. The fluidity of the freeze-dried powder was basically the same.

[0483] 1-2) The fluidity results of the dry powder inhaler showed that:

[0484] For Example 1 without adding an anticoagulant and Example 7 with leucine added as an anticoagulant, the angles of repose of the dry powder inhalers prepared were 40.2° and 38.2° respectively; the angle of repose decreased by 2.0°, and the reduction effect was significant. The powder fluidity of the dry powder inhaler was good and significant.

[0485] Comparing Example 1 and 7, the fluidity results showed that after adding an anticoagulant, the fluidity of the obtained preparation was significantly improved.

[0486] 2) Inhalation characteristics

[0487] In Example 1 without adding an anticoagulant (leucine), after the YKYY017 freeze-dried powder was directly pulverized with the fluid energy mill and then mixed with the carrier, the cohesion between the powder and the carrier was large, and the angle of repose was large. The aerodynamic mass median diameter of the drug was 3.04 μm, and the fine particle dose of the drug was 30.1%.

[0488] In Example 7 with an anticoagulant (leucine) added, after the anticoagulant and the YKYY017 freeze-dried powder were pulverized with the fluid energy mill and then mixed with the carrier, the aerodynamic mass median diameter of the drug in the obtained preparation was further reduced to 2.59 μm, and the fine particle dose of the drug increased to 42.6%. This shows that adding the anticoagulant leucine not only reduced the aggregation degree between drugs, but also improved the smoothness of the carrier surface. Compared with Example 1 (without leucine), the aerodynamic mass median diameter decreased by 0.45 μm, and the fine particle dose of the drug increased by 12.5%, showing better inhalation characteristics.

[0489] Comparing Example 1 and 7, the results of the aerodynamic mass median diameter of the drug and the fine particle dose of the drug showed that after adding an anticoagulant, the obtained preparation showed better inhalation characteristics.

[0490] 3) Delivery effect

[0491] In Example 1 without adding an anticoagulant (leucine), the delivery dose of the obtained preparation was high, and the average value accounted for 89% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the average value was distributed between 80% and 116% (required to be between 75% and 125%), and the distribution was concentrated, and the uniformity of the delivery dose was good.

[0492] Example 7: Adding 9 parts of anticoagulant (leucine), the delivery dose of the resulting preparation was further increased. The average value accounted for 97% of the labeled amount (required to be between 80% and 120%); the percentage range of the measured value to the average value was distributed between 91% and 110% (required to be between 75% and 125%), and the distribution was more concentrated, and the uniformity of the delivery dose was further significantly improved.

[0493] Comparing Example 1 and Example 7, the results of the delivery dose uniformity showed that after adding the anticoagulant, the delivery dose and the delivery dose uniformity of the resulting preparation were better.

[0494] 4) Stability

[0495] In Example 1, no anticoagulant (leucine) was added. After the resulting preparation was placed at accelerated test conditions (40°C ± 2°C, 75% ± 5% RH) for 6 months, the fine particle dose decreased from 30.1% to 28.3%, and the change rate reached 1.8%, which was less than 5%.

[0496] In Example 7, 9 parts of anticoagulant (leucine) were added. After being placed at accelerated test conditions (40°C ± 2°C, 75% ± 5% RH) for 6 months, the change rate of the fine particle dose of the resulting preparation was 1.1%. Compared with Example 1 (without anticoagulant), it showed better stability.

[0497] Summary:

[0498] 1. During the preparation process of the polypeptide dry powder inhaler, without adding an anticoagulant (for example, Example 1) or adding an anticoagulant (for example, Example 7), the fluidity, inhalation characteristics, delivery effect and stability of the prepared preparation are good. The specific manifestations are as follows:

[0499] i. The resulting preparation has a small angle of repose, controlled at about 38° - 40°, and the dry powder inhaler shows good fluidity;

[0500] ii. The aerodynamic mass median diameter of the drug in the resulting preparation is 2.59 μm - 3.04 μm, all less than 5 μm; the fine particle dose of the drug in the resulting preparation is 30.1% - 42.6%, all greater than 10%;

[0501] iii. The resulting preparation has a high and good delivery dose uniformity. The delivery dose is between 89% and 97%, all meeting the requirements (80% - 120%); the delivery uniformity is distributed between 80% and 116%, all meeting the requirements (75% - 125%);

[0502] iv. The resulting preparation has good stability. After the accelerated test, the change rate of the fine particle dose of the drug is 1.1% - 1.8%, all below 5%, and no significant change has occurred.

[0503] 2. During the preparation process of the polypeptide dry powder inhaler, compared with not adding anticoagulants, after adding anticoagulants (for example, in Example 7), the prepared preparation has better fluidity, inhalation characteristics, delivery effect and stability. The specific manifestations are as follows:

[0504] i. The angle of repose of the obtained preparation is the smallest, as low as 38.2°, and the fluidity is the best;

[0505] ii. The median aerodynamic mass diameter of the drug in the obtained preparation is small, which is 3.04 μm, less than 5 μm;

[0506] And it is reduced by 0.45 μm compared with not adding anticoagulants; the fine particle dose of the drug in the obtained preparation is large, which is 42.6%, far greater than the specified standard of 10%, and is increased by 12.5% compared with not adding anticoagulants; it shows better inhalation characteristics;

[0507] iii. The delivery dose of the obtained preparation is high and the uniformity is good. The delivery dose range is 91% - 110%, and the average delivery dose is 97% of the labeled amount, which is significantly improved compared with not adding anticoagulants (89%);

[0508] iv. The obtained preparation has better stability. After the accelerated test, the change rate of the fine particle dose of the drug is only 1.1%, less than 5%; and it is further reduced by 0.7% compared with not adding anticoagulants.

[0509] Conclusion Analysis 7: (Type of anticoagulant)

[0510] As can be seen from the results in Conclusion Analysis 6, after adding the anticoagulant leucine in Example 7, the obtained dry powder inhaler has better powder fluidity, inhalation characteristics, delivery effect and stability. Therefore, on the basis of Example 7, the type of anticoagulant added is changed to explore its influence on the inhalation characteristics and delivery effect of the dry powder inhaler. The results are as follows:

[0511] 7-1. Differences in formulation

[0512] Table 45: Differences in formulation

[0513] Example Anticoagulant type Example 7 9 parts of leucine Method 1 of Example 8 9 parts of mannitol Method 2 of Example 8 9 parts of poloxamer Example 8 Method 3 9 parts of phospholipid

[0514] 7-2. Fluidity of dry powder inhaler (Step ⑥)

[0515] Table 46: Fluidity of dry powder inhaler (Step ⑥)

[0516] Example Angle of repose (°) Example 7 38.2 Example 8 Method 1 44.3 Example 8 Method 2 46.8 Example 8 Method 3 39.9

[0517] 7-3. Inhalation characteristics of the preparation

[0518] Table 47: Inhalation characteristics of the preparation

[0519]

[0520]

[0521] 7-4. Formulation Delivery Effect

[0522] Table 48: Formulation Delivery Effect

[0523]

[0524] Conclusion:

[0525] 1) The results of fluidity show that:

[0526] The anticoagulant of Example 7 is leucine, and the angle of repose is 38.2°. The small angle of repose indicates good fluidity of the dry powder inhaler.

[0527] The anticoagulant of Method 1 of Example 8 is mannitol, and the angle of repose is 44.3°. The small angle of repose indicates good fluidity of the dry powder inhaler.

[0528] The anticoagulant of Method 2 of Example 8 is poloxamer, and the angle of repose is 46.8°. The relatively small angle of repose indicates good fluidity of the dry powder inhaler.

[0529] The anticoagulant of Method 3 of Example 8 is phospholipid, and the angle of repose is 39.9°. The small angle of repose indicates good fluidity of the dry powder inhaler.

[0530] 2) The results of inhalation characteristics show that:

[0531] The anticoagulant of Example 7 is 9 parts of leucine, and the aerodynamic mass median diameter and fine particle dose are 2.59 μm (<5 μm) and 42.6% (>10%) respectively, indicating good inhalation characteristics of the dry powder inhaler.

[0532] The anticoagulant of Method 1 of Example 8 is 9 parts of mannitol, and the aerodynamic mass median diameter and fine particle dose are 3.64 μm (<5 μm) and 39.8% (>10%) respectively, indicating good inhalation characteristics of the dry powder inhaler.

[0533] The anticoagulant of Method 2 of Example 8 is 9 parts of poloxamer, and the aerodynamic mass median diameter and fine particle dose are 3.77 μm (<5 μm) and 37.6% (>10%) respectively, indicating good inhalation characteristics of the dry powder inhaler.

[0534] The anticoagulant of Method 3 of Example 8 is 9 parts of phospholipid, and the aerodynamic mass median diameter and fine particle dose are 2.89 μm (<5 μm) and 38.7% (>10%) respectively, indicating good inhalation characteristics of the dry powder inhaler.

[0535] 3) The delivery effect shows that:

[0536] The anticoagulant in Example 7 is 9 parts of leucine. The delivery dose uniformity ranges from 91% to 110%, meeting the requirement (75% - 125%). The delivery dose is 97%, meeting the requirement (80% - 120%), indicating that the delivery dose uniformity of the dry powder inhaler is good.

[0537] The anticoagulant in Method 1 of Example 8 is 9 parts of mannitol. The delivery dose uniformity ranges from 80% to 115%, meeting the requirement (75% - 125%). The delivery dose is 95%, meeting the requirement (80% - 120%), indicating that the delivery dose uniformity of the dry powder inhaler is good.

[0538] The anticoagulant in Method 2 of Example 8 is 9 parts of poloxamer. The delivery dose uniformity ranges from 84% to 121%, meeting the requirement (75% - 125%). The delivery dose is 90%, meeting the requirement (80% - 120%), indicating that the delivery dose uniformity of the dry powder inhaler is good.

[0539] The anticoagulant in Method 3 of Example 8 is 9 parts of phospholipid. The delivery dose uniformity ranges from 92% to 113%, meeting the requirement (75% - 125%). The delivery dose is 98%, meeting the requirement (80% - 120%), indicating that the delivery dose uniformity of the dry powder inhaler is good.

[0540] Summary:

[0541] 1. Using the anticoagulants of the present invention (such as leucine, mannitol, poloxamer, and phospholipid), dry powder inhalers with good fluidity, inhalation characteristics, and delivery effects can be prepared. Specifically manifested as:

[0542] i. The resulting dry powder inhaler has a small angle of repose and good fluidity;

[0543] ii. The aerodynamic mass median diameter of the drug in the resulting preparation is 2.59 μm - 3.77 μm, less than 5 μm;

[0544] iii. The fine particle dose of the drug in the resulting preparation is 37.6% - 42.6%, far greater than 10%; the inhalation characteristics are good;

[0545] The delivery dose of the resulting preparation is 90% - 98%, meeting the limit standard (80% - 120%), and the delivery dose is high; the delivery dose uniformity ranges from 80% to 121%, and the uniformity is good.

[0546] 2. When the anticoagulants are leucine and phospholipid, the inhalation characteristics and delivery effects of the resulting preparation are better, specifically manifested as:

[0547] i. The resulting dry powder inhaler has a relatively small angle of repose and better fluidity;

[0548] ii. The median aerodynamic mass diameter of the resulting preparations is relatively small, being 2.59 μm and 2.89 μm respectively, less than 5 μm; the fine particle dose of the resulting preparations is relatively large, being 42.6% and 38.7% respectively, far greater than 10%; and the inhalation characteristics are good.

[0549] iii. The delivery doses of the resulting preparations are 97% and 98% respectively, meeting the limit standards (80% - 120%), and the delivery dose is high; the delivery dose uniformity ranges are 91% - 110% and 92% - 113% respectively, and the uniformity is good.

[0550] Conclusion Analysis 8: (Dosage of anticoagulant)

[0551] In Examples 7 and 9, dry powder inhalants were prepared using different dosages of anticoagulant, and the fluidity data of the freeze-dried powder, the fluidity of the dry powder inhalant (preparation), inhalation characteristics, delivery effect, and stability are as follows:

[0552] 8-1. Differences in the formulation of the preparation

[0553] Table 49: Differences in the formulation of the preparation

[0554] Example Anticoagulant addition amount Example 7 The amount of leucine used is 9 parts Example 9 Method 1 The amount of leucine used is 5 parts Example 9 Method 2 The amount of leucine used is 18 parts

[0555] 8-2. Fluidity of the YKYY017 freeze-dried powder (Step ⑤)

[0556] Table 50: Fluidity of the YKYY017 freeze-dried powder (Step ⑤)

[0557] Example Angle of repose (°) <![CDATA[Bulk density (g / cm 3 )]]> Example 7 50.4 0.25 Example 9 Method 1 57.1 0.19 Example 9 Method 2 48.6 0.32

[0558] 8-3. Fluidity of the dry powder inhalant

[0559] Table 51: Fluidity of the dry powder inhalant

[0560]

[0561]

[0562] 8-4. Inhalation characteristics of the dry powder inhalant

[0563] Table 52: Inhalation characteristics of the dry powder inhalant

[0564] Example Aerodynamic mass median diameter (μm) Fine particle dose (%) Example 7 2.59 42.6 Example 9 Method 1 3.91 36.3 Example 9 Method 2 2.96 39.9

[0565] 8-5. Delivery effect of the dry powder inhalant

[0566] Table 53: Delivery effect of the dry powder inhalant

[0567]

[0568] 8-6. Stability

[0569] Table 54: Stability

[0570]

[0571] Conclusion:

[0572] 1) The fluidity results of YKYY017 freeze-dried powder show that:

[0573] In Example 7, 9 parts of leucine were added as an anticoagulant. After being pulverized with YKYY017 freeze-dried powder using a jet mill, although the texture of the material was still light, leucine played an antistatic role, significantly improving the state of material agglomeration and caking. The loose bulk density increased significantly to 0.25 g / cm 3 , and the angle of repose decreased significantly to 50.4°, indicating good fluidity of YKYY017 freeze-dried powder.

[0574] In Example 9, Method 1, 5 parts of leucine were added as an anticoagulant. After being pulverized with YKYY017 freeze-dried powder using a jet mill, due to the relatively small amount of leucine added, leucine played a certain antistatic role, and the loose bulk density increased slightly to 0.19 g / cm 3 , and the angle of repose decreased slightly to 57.1°, indicating relatively good fluidity of YKYY017 freeze-dried powder.

[0575] In Example 9, Method 2, 18 parts of leucine were added as an anticoagulant. After being pulverized with YKYY017 freeze-dried powder using a jet mill, leucine played an antistatic role, and the loose bulk density increased significantly to 0.32 g / cm 3 , and the angle of repose decreased significantly to 48.6°, which was basically the same as the effect with a dosage of 9 parts.

[0576] 2) Fluidity of dry powder inhaler

[0577] In Example 7, 9 parts of leucine were added as an anticoagulant, and the angle of repose of the resulting preparation was relatively low, 38.2°, indicating good fluidity.

[0578] In Example 9, Method 1, 5 parts of leucine were added as an anticoagulant, and the angle of repose of the resulting preparation was relatively low, 44.5°, indicating relatively good fluidity.

[0579] In Example 9, Method 2, 18 parts of leucine were added as an anticoagulant, and the angle of repose of the resulting preparation was relatively low, 38.6°, indicating good fluidity, which was basically the same as the effect with a dosage of 9 parts.

[0580] 3) The inhalation characteristic results show

[0581] In Examples 7 and 9, different amounts of anticoagulant (leucine) were added. After directly pulverizing the YKYY017 freeze-dried powder with a pneumatic grinder and mixing it with the carrier, leucine not only reduced the degree of aggregation between drugs, but also improved the smoothness of the carrier surface, so that the drug was more likely to disperse from the carrier during the administration process, showing good inhalation characteristics. Moreover, after the samples were placed for 6 months under accelerated conditions, the change rate of the fine particle dose was smaller, indicating better stability of the samples.

[0582] In Examples 7 and 9 (Method 1, Method 2), when the amounts of leucine were 9 parts, 5 parts, and 18 parts respectively, the aerodynamic mass median diameters of the drug were 2.59 μm, 3.91 μm, and 2.96 μm respectively, and the fine particle doses were 42.6%, 36.3%, and 39.9% respectively. Compared with Example 1 (without leucine), better inhalation characteristics were shown.

[0583] 4) The delivery effect shows that:

[0584] In Examples 7 and 9, different amounts of anticoagulant (leucine) were added. After directly pulverizing the YKYY017 freeze-dried powder with a pneumatic grinder and then mixing it with the carrier, the delivery dose was more uniform, indicating that after adding the anticoagulant (leucine), the materials were easier to mix evenly or disperse from the blister or capsule.

[0585] In Examples 7 and 9 (Method 1, Method 2), when the amounts of leucine were 9 parts, 5 parts, and 18 parts respectively, the delivery doses were between 91% - 110%, 84% - 115%, and 92% - 116% respectively, and the average delivery doses were 97%, 93%, and 99% of the labeled amount respectively. Compared with Example 1, the uniformity of the delivery dose was better, further indicating that the differences in the obtained preparations were small, the quality was more uniform, and the accuracy of the administration dose was improved.

[0586] 5) The stability results show that:

[0587] After being placed for 6 months under accelerated test conditions (40°C ± 2°C, 75% ± 5% RH), compared with Example 1, in Examples 7 and 9, different amounts of anticoagulant (leucine) were added, and the change rate of the fine particle dose of the obtained preparations decreased significantly, indicating that after adding the anticoagulant (leucine), the obtained preparations had better stability.

[0588] In Examples 7 and 9 (Method 1, Method 2), when the amounts of leucine were 9 parts, 5 parts, and 18 parts respectively, the change rates of the fine particle dose were 1.1%, 1.6%, and 1.0% respectively. Compared with Example 1 (1.8%), better stability was shown.

[0589] Summary:

[0590] 1. During the process of preparing the COVID-19 polypeptide dry powder inhaler, adding 5 to 18 parts of anticoagulant can improve the quality of the preparation, and the obtained preparation has good fluidity, inhalation characteristics, delivery effect and stability.

[0591] Specifically manifested as:

[0592] i. The obtained preparation has good fluidity, with a small angle of repose, only 38.2° - 44.5°, less than 45°.

[0593] ii. The aerodynamic mass median diameter of the obtained preparation is less than 5 μm, being 2.59 μm - 3.91 μm; the fine particle dose of the drug in the obtained preparation is high, being 36.3% - 42.6%, far greater than 10%.

[0594] iii. The obtained preparation has a high and good uniformity of delivery dose, with the delivery dose up to 93% - 99%; the uniformity range of the delivery dose is distributed between 84% - 116%, with a narrow distribution range, showing good delivery dose uniformity.

[0595] iv. The obtained preparation has good stability, and the change rate of the fine particle dose of the drug in the obtained preparation is between 1.0% - 1.6%, far less than 5%, and no significant changes have occurred.

[0596] Conclusion analysis 9: (Control examples 1 and 2)

[0597] In control example 1, the drug YKYY017 in the present invention was made into a dry powder inhaler according to the prescription (type) in example 9 of patent CN102264365A; in control example 2, the drug YKYY017 in the present invention was made into a dry powder inhaler according to the prescription in example 7 of patent CN114727969A. The present invention (example 1 with better effect without adding anticoagulant, method 1 of example 5 with worse effect without adding anticoagulant, and example 7 with added anticoagulant) was compared with control example 1 and control example 2 to explore the influence on the fluidity, inhalation characteristics, delivery effect and stability of the YKYY017 dry powder inhaler, and the results are as follows:

[0598] 9 - 1. Differences in preparation prescriptions

[0599] Table 55: Differences in preparation prescriptions

[0600] Example Prescription Example 1 Drug active ingredient, excipient, stabilizer, carrier Example 5 Method 1 Drug active ingredient, excipient, stabilizer, carrier Example 7 Drug active ingredient, excipient, stabilizer, carrier, anticoagulant Control Example 1 Drug active ingredient, carrier, stabilizer Control Example 2 Drug active ingredient, excipient, solvent

[0601] 9 - 2. Fluidity of dry powder inhaler

[0602] Table 56: Fluidity of dry powder inhaler

[0603] Example Angle of repose (°) Example 1 40.2 Example 5 Method 1 43.1 Example 7 38.2 Control Example 1 58.6 Control Example 2 61.3

[0604] 9-3. Inhalation characteristics

[0605] Table 57: Inhalation characteristics

[0606] Example Aerodynamic mass median diameter (μm) Fine particle dose (%) Example 1 3.04 30.1 Example 5 Method 1 3.56 27.5 Example 7 2.59 42.6 Control Example 1 7.36 22.5 Control Example 2 8.98 15.6

[0607] 9-4. Delivery effect

[0608] Table 58: Delivery effect

[0609]

[0610] 9-5. Stability

[0611] Table 59: Stability

[0612]

[0613] Conclusion:

[0614] 1) The results of fluidity show that:

[0615] The angle of repose of Example 1, Method 1 of Example 5 and Example 7 are 40.2°, 43.1° and 38.2°, respectively, which are less than 45°, indicating good fluidity. While the angles of repose of Comparative Example 1 and Comparative Example 2 are 58.6° and 61.3°, respectively, which are significantly higher than 45°, indicating poor fluidity.

[0616] 2) The results of inhalation characteristics show that:

[0617] The aerodynamic mass median diameters of the drugs in Example 1, Method 1 of Example 5 and Example 7 are 3.04μm, 3.56μm and 2.59μm, respectively, and the fine particle doses are 30.1%, 27.5% and 42.6%, respectively; the inhalation characteristics are good.

[0618] The aerodynamic mass median diameters of the drugs in Comparative Example 1 and Comparative Example 2 are 7.36μm and 8.98μm, respectively, which are significantly higher than those of Example 1, Method 1 of Example 5 and Example 7 of this application; the fine particle doses are 22.5% and 15.6%, respectively, which are significantly lower than those of Example 1, Method 1 of Example 5 and Example 7 of this application, and the inhalation characteristics are lower than those of this application.

[0619] 3) The delivery effect shows that:

[0620] The delivery doses of Example 1, Method 1 of Example 5 and Example 7 are 83% - 97%, with high delivery doses; the uniformity range of the delivery dose is controlled within 80% - 124%, with a narrow range; and it is significantly better than the limit standard.

[0621] The delivery doses of Comparative Example 1 and Comparative Example 2 were reduced to 77% and 81%, respectively, with low delivery doses; the delivery dose uniformity increased to 77% - 136%, with a significantly wider range than that of Example 1, Method 1 of Example 5, and Example 7, and poor delivery dose uniformity.

[0622] 4) The stability results showed that:

[0623] Example 1, Method 1 of Example 5, and Example 7 were placed at (25°C ± 2°C, 75% ± 5% RH) under accelerated test conditions for 6 months. Compared with day 0, the change rates of the drug active ingredient were 0.5%, 2.4%, and 0.1%, respectively. Under the same conditions, the change rates of the drug active ingredient in Comparative Example 1 and Comparative Example 2 increased significantly to 18.7% and 21.9%, respectively, with poor stability and unable to prepare qualified dry powder inhalants. The above results indicate that the compositions and preparation methods provided by the present invention (Example 1, Method 1 of Example 5, and Example 7) can significantly improve the stability of YKYY017.

[0624] Summary:

[0625] 1. When preparing dry powder inhalants using the prescription of the present invention, the resulting preparations (e.g., Example 1, Method 1 of Example 5, and Example 7) have good fluidity, good inhalation characteristics, delivery effects, and stability. Specifically manifested as:

[0626] i. The angle of repose of the resulting preparations is controlled within 38.2° - 43.1°, < 45°, indicating good material fluidity;

[0627] ii. The aerodynamic mass median diameter of the drug in the resulting preparations is relatively small, only about 2.5 - 3.5 μm,

[0628] far less than the limit standard of 5 μm;

[0629] The fine particle dose of the drug is relatively large, 27.5% - 42.6%, far higher than the limit standard of 10%;

[0630] iii. The delivery dose difference of the resulting preparations is small and the delivery dose is relatively high, up to 83% - 97%; the delivery dose uniformity range is distributed within 80% - 124%, with a narrow distribution range and high delivery dose uniformity;

[0631] iv. The resulting preparations have good stability. After accelerated testing, the change rate of the drug active ingredient content is only 0.1% - 2.4%, far less than 5%.

[0632] 2. When preparing dry powder inhalants using other ingredient prescriptions (e.g., Comparative Example 1 and Comparative Example 2), the resulting preparations have poor fluidity, and the inhalation characteristics, delivery effects, and stability are significantly reduced. Specifically manifested as:

[0633] i. The angle of repose of the obtained preparation is large, being 58.6° and 61.3°, greater than 45°, and the fluidity of the material is poor.

[0634] ii. The aerodynamic mass median diameter of the drug in the obtained preparation is large, being 7.36 μm and 8.98 μm.

[0635] The dose of fine drug particles is low, being 22.5% and 15.6%, slightly greater than 10%.

[0636] iii. The difference in the delivery dose of the obtained preparation is large and the delivery dose is low. The delivery doses are 77% and 81%.

[0637] The range of the delivery dose uniformity is distributed from 77% to 136%, the distribution range is too wide, the uniformity is poor, and it does not meet the specified distribution range (75% - 125%).

[0638] iv. The stability of the obtained preparation is poor. After the accelerated test, the change rates of the content of the drug active ingredient are relatively large, being 18.7% and 21.9% respectively, far greater than 5%.

Claims

1. A dry powder inhaler, which comprises a pharmaceutically active ingredient, a stabilizer, an excipient, a carrier and an anticoagulant, wherein, the pharmaceutically active ingredient comprises a polypeptide; the polypeptide comprises YKYY017, and the peptide sequence of YKYY017 is shown as SEQ ID NO.1: Ac-Ser-Val-Val-Asn-Ile-Gln-Lys-Glu-Ile-Asp-Arg-Leu-Asn-Glu-Val-Ala-Lys-Asn-Leu-Asn-Glu-Ser-Leu-Ile-Asp-Leu-Gln-Glu-Leu-Gly-Lys-Tyr-Glu-Gln-Tyr-Ile-Lys-Glu-Ala-Ala-Ala-Lys-Lys(Chol)-NH 2 ; the stabilizer is a buffered saline solution with a pH of 3.0 to 4.0; the stabilizer comprises: acetic acid - acetate solution and / or citric acid - citrate solution; the excipient is selected from one or more of the following substances: mannitol, sucrose, trehalose; the carrier is lactose and / or mannitol; the anticoagulant is leucine and / or phospholipid; by mass, the dosage of the pharmaceutically active ingredient is 5 to 15 parts, the dosage of the excipient is 2 to 8 parts, the dosage of the carrier is 15 to 50 parts, and the dosage of the anticoagulant is 5 to 18 parts; the particle size of the carrier is between 1 μm and 250 μm, wherein D50 is 80 μm to 120 μm.

2. The dry powder inhaler according to claim 1, wherein, the stabilizer comprises: acetic acid - acetate solution with a pH of 3.0 to 4.

0.

3. The dry powder inhaler according to claim 1 or 2, wherein, the pH of the stabilizer is 3.

7.

4. The dry powder inhaler according to claim 1 or 2, wherein, the dosage of the carrier is 30 to 35 parts.

5. The dry powder inhaler according to claim 1 or 2, wherein, the dosage of the anticoagulant is 8 to 12 parts.

6. The dry powder inhaler according to claim 1 or 2, wherein, the synthesis method of the polypeptide is as follows: a) Select a resin and couple all amino acids in sequence from the C-terminus to the N-terminus to synthesize a fully protected polypeptide by solid-phase synthesis; b) Remove the side-chain protecting group of Lys; c) Couple cholesterol succinate monoester to the side chain in the presence of a coupling agent; d) In the presence of a cleavage agent, cleave the peptide resin and deprotect the side chain to obtain the polypeptide.

7. The dry powder inhaler according to claim 6, wherein, in step a), the resin used is Rink Amide - MBHA Resin.

8. The dry powder inhaler according to claim 6, wherein, in step b), hydrazine is used as a catalyst to selectively remove the side-chain protecting group.

9. The dry powder inhaler according to claim 1 or 2, wherein, the dry powder inhaler is administered by oral inhalation using a Turbuhaler.

10. The dry powder inhaler according to claim 1 or 2, wherein, the excipient is mannitol.

11. The dry powder inhaler according to claim 1 or 2, wherein, the carrier is lactose.

12. The dry powder inhaler according to claim 1 or 2, wherein, the anticoagulant is leucine.

13. A preparation method of the dry powder inhaler according to any one of claims 1 to 12, comprising the following steps: ① Prepare the stabilizer; ② Dissolve the excipient in the stabilizer; ③ Dissolve the pharmaceutically active ingredient in the system of step ②, and adjust the pH to obtain a solution; ④ Freeze-dry the solution obtained in step ③ to obtain a lyophilized powder; ⑤ Grind; ⑥ Mix: Mix the ground powder with the carrier and grind to obtain the dry powder inhaler.

14. The preparation method according to claim 13, wherein, the preparation method further includes mixing an anticoagulant with the lyophilized powder and pulverizing the mixture in step ⑤.

15. The preparation method according to claim 13 or 14, wherein, in step ⑤, the D90 of the pulverized particles is below 10 μm.

16. The preparation method according to claim 15, wherein, in step ⑤, the D90 of the pulverized particles is 1.0 μm to 5.0 μm.

17. The preparation method according to claim 13 or 14, wherein, in step ⑥, the carrier particle size is between 1 μm and 250 μm, and the D50 is 80 μm to 120 μm.

18. The preparation method according to claim 13 or 14, wherein, after freeze-drying in step ④, the water content is controlled to be ≤ 1.0%.

19. Use of the dry powder inhaler according to any one of claims 1 to 12 in the preparation of a medicament for treating coronavirus infection.

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