Inhalation dry powder preparation containing salmeterol and tiotropium salt composition and preparation method thereof

Abstract

The invention relates to an inhalation dry powder preparation containing a salmeterol and tiotropium salt composition and a preparation method of the inhalation dry powder preparation, and belongs to the technical field of pharmaceutical preparations. The preparation comprises a long-acting beta-2 adrenergic receptor agonist (LABA) salmeterol and a long-acting anticholinergic drug (LAMA) tiotropium salt, and the efficient synergistic effect of the two drugs is achieved by optimizing a drug micronization process, a carrier particle design, a carrier material and a preparation process. The medicine can penetrate into a lung target region, so that precise treatment is realized. A powder mist inhalation device CN219423486U disclosed by Nanchang large pharmaceutical industry limited company is adopted as a delivery device, and the lung deposition rate and delivery efficiency of the medicine are remarkably improved by controlling the particle size of the medicine, the particle property of a carrier and the uniformity of the preparation. In addition, the invention further provides a preparation method of the preparation, the preparation method comprises the processes of medicine micronization, mixing homogenization and packaging, and the stability and dosage accuracy of the preparation are ensured. Experiments show that in an accelerated stability investigation period as long as 6 months, the drug content, impurity level and fine particle fraction in the preparation are kept stable and do not change significantly, and it is proved that the preparation has good physical and chemical stability and storage performance. The preparation has quick-acting and long-acting bronchiectasia effects, is suitable for long-term management of moderate and severe asthma and chronic obstructive pulmonary disease (COPD), and can significantly improve the medication convenience and treatment compliance of patients.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and in particular to an inhalation dry powder preparation containing a salmeterol and tiotropium salt composition and a preparation method thereof. Background Art

[0002] Salmeterol is a long-acting β2 receptor agonist (LABA), commonly used to treat bronchial asthma and chronic obstructive pulmonary disease. It can relax bronchial smooth muscles and dilate airways by activating β2 receptors in the lungs, thereby alleviating asthma symptoms such as dyspnea and wheezing. Compared with other β2 receptor agonists, salmeterol has a longer half-life, so its efficacy can last for 24 hours. It also has high selectivity, acting only on β2 receptors in the bronchi and lungs, and will not affect the β1 receptors in the heart, so there are fewer adverse cardiovascular reactions.

[0003] Tiotropium bromide is a new second-generation long-acting anticholinergic drug (LAMA). As a selective bronchodilator, it can specifically antagonize the M1-M5 cholinergic receptors on bronchial smooth muscle. Its binding ability to cholinergic receptors is 10 times that of traditional ipratropium bromide, so it can quickly relax bronchial smooth muscle and effectively relieve dyspnea symptoms. In addition, tiotropium bromide has a strong and lasting bronchodilator effect and a long half-life. It can improve bronchoconstriction symptoms at night and keep the airway open all day, effectively preventing repeated collapse and opening of the airway and friction of the respiratory tissue.

[0004] Moderate to severe asthma and chronic obstructive pulmonary disease (COPD) are diseases characterized by chronic inflammation, airway obstruction, and airway hyperresponsiveness. They are difficult to treat and have a higher mortality rate than patients with single asthma and chronic obstructive pulmonary disease (COPD). Bronchodilators, antibiotics, anticholinergics, etc. are often used for clinical treatment, but long-term use is prone to problems such as airway fungal infection. Tiotropium bromide is a commonly used drug for stable COPD, which can relax the bronchi, relieve cough and shortness of breath symptoms, and thus improve the patient's lung function and inhibit airway inflammation. However, tiotropium bromide cannot prevent airflow limitation, and long-term use can cause drug resistance and adverse reactions. Studies have shown that salmeterol can be used to treat ACOS and has certain safety. In addition, existing single-drug inhalation preparations have certain limitations in terms of therapeutic effect and patient compliance, especially for moderate to severe patients, where a single drug is often difficult to fully control symptoms. Therefore, the development of composite inhalation preparations with multiple mechanisms of action has become a research direction.

[0005] It has been found that unexpected therapeutic benefits, especially synergistic therapeutic benefits, can be obtained in the treatment of inflammatory or obstructive airway diseases by combined treatment with salmeterol or its salt or solvate and tiotropium salt. For example, compared with the treatment with salmeterol or tiotropium salt alone, the use of this combined therapy can significantly reduce the dose required to achieve a given therapeutic effect, thereby greatly reducing possible adverse side effects. On the other hand, the combined inhalation of bronchodilators with different mechanisms of action can enhance the bronchodilator effect of the drug, effectively improve the hyperinflation and dyspnea of ​​COPD patients, and significantly improve various lung function indicators. Studies have shown that the combined use of tiotropium bromide and salmeterol can reduce the levels of peripheral blood eosinophils (EOS) and total immunoglobulin E (IgE), indicating that combined treatment can reduce the body's allergic reactivity. Salmeterol specifically binds to β2 receptors to relieve bronchospasm, accelerate blood circulation, and enhance the body's defense system. Its combined action with tiotropium bromide can block the generation of inflammatory cells, reduce airway and lung inflammatory reactions, and thus improve airway hyperresponsiveness. Salmeterol combined with tiotropium bromide in the treatment of ACOS can improve clinical efficacy, improve lung function, and relieve airway inflammatory response.

[0006] At present, some studies have explored the combined use of LABA and LAMA, such as the combination of salmeterol and tiotropium. However, existing inhalation preparations still face many challenges in terms of drug stability, dosage accuracy, delivery efficiency, and patient convenience. For example, the physicochemical properties of different drugs may lead to decreased preparation uniformity or insufficient storage stability; differences in inhalation devices can also affect the pressure drop value and lung deposition rate of the drug. In addition, the existing technology has not yet fully addressed how to reduce drug deposition in non-targeted sites while delivering high doses, thereby reducing the risk of side effects.

[0007] Therefore, the development of a compound inhalation preparation based on salmeterol and tiotropium bromide requires not only the optimization of drug ratio and preparation process, but also the combination of advanced inhalation device design to ensure efficient drug delivery and long-term stability. This preparation is expected to provide a better treatment option for patients with moderate to severe respiratory diseases, while improving patient compliance and quality of life. Summary of the invention

[0008] In order to overcome the defects of the prior art and meet the needs of the art, the purpose of the present invention is to provide a dry powder composition containing salmeterol or a pharmaceutically acceptable salt thereof and tiotropium salt for oral inhalation administration, which can accurately and uniformly deliver the set dose of each active ingredient to the effective site, so that the two active ingredients can synergistically play the role of relaxing bronchial smooth muscles.

[0009] Another object of the present invention is to provide a method for preparing the dry powder pharmaceutical composition.

[0010] A further object of the present invention is to provide uses of the dry powder pharmaceutical composition.

[0011] The present invention adopts the following technical solution to solve the problem:

[0012] The invention provides an inhalable dry powder composition of medicine, which consists of salmeterol or a pharmaceutically acceptable salt thereof, tiotropium bromide and a pharmaceutically acceptable carrier.

[0013] The invention is characterized in that suitable pharmaceutically acceptable salts of salmeterol include inorganic acid salts or organic acid salts known in the art, and salmeterol xinafoate is particularly preferred.

[0014] The salmeterol is prepared by Friedel-Crafts acylation reaction of 2-hydroxybenzaldehyde and bromoacetyl bromide to generate an intermediate (I), reaction of 4-phenyl-1-butanol and 1,6-dibromohexane to generate an intermediate (II), separation, crystallization and extraction to obtain an intermediate (III), condensation of the intermediate (I) and the intermediate (III) followed by reduction catalytic hydrogenation and debenzylation to obtain the salmeterol (VI).

[0015] The suitable pharmaceutically acceptable salt of salmeterol is prepared by reacting salmeterol with an acid.

[0016] The tiotropium bromide is prepared by reacting 2,2-dithienyl glycolic acid methyl ester with scopolol through an ester exchange reaction to obtain 2,2-dithienyl glycolic acid scopolyl; the separated liquid is acidified with dilute hydrochloric acid, washed with toluene, and alkalized with sodium carbonate acid solution; the separated liquid is extracted with dichloromethane, washed three times with purified water, dried and filtered after adding anhydrous sodium sulfate, and then recrystallized with acetonitrile, filtered and dried to obtain 2,2-dithienyl glycolic acid scopolyl; 2,2-dithienyl glycolic acid scopolyl is added with acetonitrile and dichloromethane solvent to react with methyl bromide at low temperature through methylation and bromination, and filtrated under reduced pressure to obtain a crude tiotropium bromide; the crude tiotropium bromide is synthesized from methyl bromide; the crude tiotropium bromide is decolorized and filtered by heating with activated carbon, cooled for crystallization, and then recrystallized with acetonitrile-methanol, and dried under reduced pressure and vacuum to obtain tiotropium bromide.

[0017] The 2,2-dithienyl glycolic acid methyl ester and scopolamine undergo an ester exchange reaction to obtain intermediate III, as shown in the schematic diagram. Figure 1 .

[0018] The intermediate III is synthesized with methyl bromide to obtain crude tiotropium bromide, as shown in the schematic diagram. Figure 2 .

[0019] The pharmaceutically acceptable carrier can be selected from one or more of lactose, sucrose, mannitol, and amino acids, wherein the amino acids include glycine, alanine, valine, leucine, isoleucine, methionine (methionine), proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine and histidine; glycine is particularly preferred.

[0020] The preparation method of the dry powder composition of the present invention comprises the following steps:

[0021] Step 1) Synthesis of salmeterol: 2-Hydroxybenzaldehyde and bromoacetyl bromide undergo Friedel-Crafts acylation to generate intermediate (I), 4-phenyl-1-butanol and 1,6-dibromohexane react to generate intermediate (II), separation, crystallization and extraction are performed to obtain intermediate (III), intermediate (I) and intermediate (III) undergo condensation and then reduction catalytic hydrogenation and debenzylation to obtain salmeterol (VI).

[0022] Step 2) reacting salmeterol with an acid to obtain a pharmaceutically acceptable salt of salmeterol.

[0023] Step 3) Synthesis of Tiotropium Bromide: In an anhydrous and oxygen-free box, control the temperature to ≤15°C; evacuate the reaction bottle, fill it with nitrogen, and then add 2,2-dithienyl glycolic acid methyl ester, scopolamine, anhydrous toluene, and metal sodium tablets; after the addition is completed, turn on the heating, open the vacuum, and control the liquid temperature to 70-78°C for reaction; the reaction solution is acidified by dilute hydrochloric acid, washed with toluene, and alkalized with sodium carbonate acid; then extract the solution with dichloromethane, wash it with purified water three times, and add anhydrous The product is dried over sodium sulfate and filtered, and then recrystallized through acetonitrile. After filtering and drying, 2,2-dithienyl glycolic acid scopolamine is obtained; 2,2-dithienyl glycolic acid scopolamine is added with acetonitrile and dichloromethane solvent to react with methyl bromide at low temperature through methylation and bromination, and the crude product of tiotropium bromide is obtained by filtration under reduced pressure; the crude product of tiotropium bromide is synthesized from methyl bromide; the crude product of tiotropium bromide is decolorized by heating with activated carbon, filtered, cooled and crystallized, and then recrystallized from acetonitrile-methanol, and dried under reduced pressure and vacuum to obtain tiotropium bromide.

[0024] The tiotropium bromide is tested according to the European Pharmacopoeia EP5, (1) impurity A: ≤0.15% (2) impurity B: ≤0.10% (3) impurity C: ≤0.1% (4) impurity D: ≤0.1% (5) impurity E: ≤0.1% (6) impurity F: ≤0.1% (7) single unknown impurity: ≤0.10%; total impurities ≤0.3%.

[0025] Step 4) Micronization of salmeterol or its pharmaceutically acceptable salt: Control the humidity of the operating environment to 25-45%, preferably 30%. Micronization can be performed by air flow milling, high-speed grinding, or ball milling.

[0026] Step 5) Micronization of tiotropium bromide: Control the humidity of the operating environment to 25-45%, preferably 30%. Micronization can be performed by air flow milling, high-speed grinding, or ball milling.

[0027] Step 6) Detect the particle size of the ultrafine powder of salmeterol or its pharmaceutically acceptable salt, ultrafine powder of tiotropium salt, and ultrafine powder of pharmaceutically acceptable carrier. If the powder passes the test, weigh the powder according to the prescribed amount for later use.

[0028] Step 7) Preparation of a pharmaceutically acceptable carrier: Control the humidity of the operating environment to 25-45%, preferably 30%, pass ≥ 90% of the pharmaceutically acceptable carrier crystals that cannot pass through an 80-mesh sieve through a hammer mill equipped with a 0.2 mm screen, and grind to obtain a pharmaceutically acceptable carrier powder; place the pharmaceutically acceptable carrier powder in an ultrasonic vibration sieve equipped with a 90 um screen, and ultrasonically screen to obtain a pharmaceutically acceptable carrier fine powder; place the pharmaceutically acceptable carrier fine powder in an airflow mill, and grind to obtain a pharmaceutically acceptable carrier ultrafine powder.

[0029] The above steps can be replaced by purchasing commercially available inhaled lactose, inhaled glycine or inhaled mannitol that meet the particle size control requirements.

[0030] Step 8) Controlling the humidity of the operating environment to 25-45%, preferably 30%, the salmeterol or a pharmaceutically acceptable salt thereof, tiotropium salt, and a pharmaceutically acceptable carrier are mixed by one or more of the following methods: sieving mixing, V-shaped mixing, three-dimensional mixing, shear mixing, etc., preferably shear mixing.

[0031] The composition is mixed, and the weight ratio of tiotropium salt: salmeterol or its pharmaceutically acceptable salt: pharmaceutically acceptable carrier is 1:1-15:500-50000, preferably 1:3:550.

[0032] Step 9) The mixed composition is loaded into a gelatin hollow capsule or a hypromellose capsule by quantitative cannula filling or vacuum drum filling, preferably a hypromellose capsule, with a filling amount of 5 to 35 mg.

[0033] Step 10) The filled capsules are packaged with aluminum-plastic packaging to prepare commercially available packaging products.

[0034] The capsule content composition is administered by oral inhalation using an inhalation device and delivered to the lungs. Preferably, a powder mist inhalation device CN219423486U disclosed by Nanchang Hongyi Pharmaceutical Co., Ltd. is used. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Attached Figure 1 This is a schematic diagram of the ester exchange reaction between 2,2-dithienyl glycolic acid methyl ester and scopolol to obtain intermediate III.

[0036] Attached Figure 2This is a schematic diagram of the synthesis of crude tiotropium bromide from intermediate III and methyl bromide. DETAILED DESCRIPTION

[0037] The present invention is further described in detail below in conjunction with specific examples. The examples given are only for illustrating the present invention, rather than for limiting the scope of the present invention.

[0038] The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer. Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art.

[0039] In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0040] All fine particle quantity and delivery uniformity tests were conducted using a powder mist inhalation device CN219423486U disclosed by Nanchang Hongyi Pharmaceutical Co., Ltd. unless otherwise specified.

[0041] Example 1: Synthesis of Salmeterol Xinafoate

[0042] Salmeterol is obtained through six steps of reaction, and the impurities can be controlled to meet the requirements of the United States Pharmacopoeia. The total impurities of salmeterol do not exceed 0.9%. [Note: The total impurities are calculated from the sum of all impurity peaks greater than or equal to 0.05%.]

[0043] Synthesis of 5-(bromoacetyl)-2-hydroxybenzaldehyde (intermediate I): In anhydrous aluminum chloride and anhydrous dichloromethane system, bromoacetyl bromide and 2-hydroxybenzaldehyde were added dropwise at low temperature and refluxed, then quenched with ice water, extracted, washed with saturated brine, dried and concentrated, and recrystallized from toluene to obtain a yellow solid.

[0044] Synthesis of [4-(6-bromohexyloxy)]butylbenzene (Intermediate II): 4-phenyl-1-butanol, 1,6-dibromohexane, potassium hydroxide and tetrabutylammonium bromide (TBAB) were reacted at room temperature for 24 hours, and then filtered, dissolved in ether, washed with water, dried and distilled under reduced pressure to obtain a colorless liquid.

[0045] Synthesis of N-benzyl-6-(4-phenylbutoxy)hexylamine (Intermediate III): After the intermediate II reacts with benzylamine at 120°C for 8 hours, the excess benzylamine is removed by vacuum distillation, and the product is separated by acidification with hydrobromic acid, recrystallization with chloroform, and purification by alkaline ether extraction to finally obtain a light yellow oil.

[0046] Synthesis of 2-hydroxy-5-{[6,6-(4-phenylbutoxy)hexyl]benzyl]amino}acetylbenzaldehyde (Intermediate IV): Intermediate I and Intermediate III were added dropwise in the presence of acetonitrile and potassium hydroxide at 50°C and refluxed for 1 hour. After cooling and evaporation to remove the solvent, the mixture was directly used in the next step.

[0047] Synthesis of 2-hydroxy-5-[[2-[[6,6-(4-phenylbutoxy)hexylbenzyl]amino]-1-hydroxy]ethylbenzyl alcohol (Intermediate V): Intermediate IV was reduced with an ethanol solution of sodium borohydride for 15 hours, then acidified and concentrated, extracted with alkali (chloroform), washed with water and dried to obtain the crude product Intermediate V.

[0048] Synthesis of salmeterol: The intermediate V and methanol are hydrogenated at room temperature and pressure for 36 hours. After the reaction is completed, the yellow oil is filtered and concentrated to obtain a white solid, which is then stirred with toluene to precipitate.

[0049] Synthesis of salmeterol xinafoate: ethanol and intermediate VI are added to a reaction bottle in sequence, stirred to dissolve; xinafoate is added, stirred to dissolve, cooled to crystallize, filtered, and the filter cake is dried to obtain the product.

[0050] The salmeterol xinafoate was tested according to the United States Pharmacopoeia, and all test standards were qualified, and the qualified products were kept for future use.

[0051] Determination of related substances in salmeterol xinafoate by HPLC: Using the United States Pharmacopoeia liquid chromatography conditions, the salmeterol xinafoate sample was analyzed and determined at a wavelength of 278nm, and the total impurities were calculated to be 0.1%.

[0052] Example 2: Synthesis of Tiotropium Bromide

[0053] In an anhydrous and oxygen-free box, control the temperature to ≤15°C; evacuate a 3000ml four-necked reaction bottle and fill it with nitrogen to normal pressure. Under nitrogen protection at 1-2L / min (nitrogen protection), add 100g of methyl 2,2-dithienyl glycolate, 164g of scopolamine, 1600ml of anhydrous toluene, and about 7.2g of metal sodium tablets.

[0054] Start stirring until it forms a vortex, start heating, start vacuum, control the liquid temperature at 70-78°C and the vacuum at -0.06-0.08Mpa to react, promptly remove the methanol generated by the reaction, and distill the dripped toluene methanol distillate.

[0055] Add an appropriate amount of anhydrous toluene from the dropping funnel and keep the amount of dripping basically balanced. Raise the liquid temperature to 65℃ and time the reaction. Keep a large number of bubbles in the reaction for 1 hour. The reaction is completed in about 6 hours. Switch to an ice water bath and cool to room temperature with reduced pressure and stirring.

[0056] Take the toluene reaction solution, slowly add it to 2400ml of 5% hydrochloric acid solution under stirring until the end point pH is 1-2, pour the acidified solution into a 5000ml glass separatory funnel, shake well, and let it stand for stratification. Let it stand for about 20 minutes (about 20 minutes) until the stratification is clear, and release the lower layer of acid water from the drain valve. Pour the acid water into a 5000ml glass separatory funnel, add 730ml of toluene, shake well, and let it stand for about 10 minutes until the stratification is clear. Release the lower layer of acid water from the drain valve, and wash it twice more in the same way (wash 3 times in total); slowly add 3000ml of saturated sodium carbonate solution to the container containing the acid water until the end point pH is 9-10, and the alkaline solution is obtained.

[0057] Add 1130ml of dichloromethane to the alkalized solution, stir until it is clear, pour it into a 10000ml glass separatory funnel, shake it well and let it stand for about 5 minutes until the layers are clear. Release the lower dichloromethane extract from the drain valve. Extract the upper water liquid twice more in the same way (a total of 3 extractions), collect the dichloromethane extract, and combine it with the first extract.

[0058] Add dichloromethane extract to 500ml purified water, shake well, and let stand for about 5 minutes until the layers are clear. Drain the lower layer of dichloromethane extract from the drain valve. Wash the dichloromethane extract twice more in the same way (3 times in total).

[0059] Add about 1300g of anhydrous sodium sulfate into the container filled with dichloromethane liquid, seal it and let it stand for dehydration and drying for 14 hours.

[0060] Add about 428 ml of acetonitrile to the evaporating bottle of the rotary evaporator, set a water bath at about 80°C (about 80°C) and rotate to dissolve. Use a stainless steel funnel filter with 4 layers (2-4 layers) of filter paper to reduce pressure and filter, and wash the crystallized filter cake with about 286m ≤0°C acetonitrile twice and drain.

[0061] Take the wet product and dry it in a vacuum drying oven at 40℃ and -0.08MP (vacuum) for 2h, turning the material over once. Then heat it to 80℃ and dry it at -0.08MP (vacuum) for 8h to obtain 2,2-dithienyl glycolic acid scopolamine, weighing about 97.5g

[0062] Add about 96g of scopolamine 2,2-dithienyl glycolate and about 960ml of acetonitrile-dichloromethane (1:1.1) solution to the rotary evaporator bottle. Set to about 40℃ and rotate until dissolved. Remove the rotary evaporator bottle, add a magnetic stirrer, seal it, and cool it from -20 to 0℃ to ≤0℃.

[0063] Take about 139g of sealed methyl bromide at ≤0℃ and quickly pour it into the acetonitrile-dichloromethane (1:1.1) solution in the rotary evaporation bottle, immediately seal it with a rubber stopper and wrap it with tape. Place the solution in the sealed rotary evaporation bottle in a water bath on a magnetic stirrer, stir it in a vortex shape, and keep it at 25℃ for 48h.

[0064] Take a rotary evaporation bottle and use a stainless steel filter equipped with 2 layers of medium-speed filter paper to filter under reduced pressure to obtain the mother liquor of the synthetic crude product.

[0065] Add about 120g of crude mother liquor, add about 240ml of 50% methanol solution, install a reflux pipe to pass cooling water, heat to 60-65℃ and stir until dissolved, add about 18g of activated carbon, stir and reflux at 60-65℃ to decolorize for 0.5h.

[0066] Preheat the stainless steel funnel filter with two layers of filter paper to about 80°C in a drying oven, install it on a glass suction bottle, quickly pour in the decolorizing solution and filter under reduced pressure. Wash twice with about 48 ml of acetonitrile-methanol solution (9:1) and drain.

[0067] Install a stainless steel funnel filter with two layers of filter paper on a glass suction flask, pour in the crystallization solution and filter under reduced pressure. Wash the crystals twice with about 240 ml of ≤0℃ acetonitrile-methanol solution (9:1), and drain. Get the recrystallized wet product.

[0068] Take the recrystallized wet product and dry it in a vacuum drying oven at 40℃ -0.08MP (vacuum) for 2h, turning the material once. Then heat it to 105℃ -0.08MP (vacuum) and dry it for 16h. Turn the material once when drying at 105℃ for about 2h to obtain the finished product of tiotropium bromide, weighing about 60.0g.

[0069] The finished product is sampled for testing and vacuum packed.

[0070] The finished product of tiotropium bromide was tested using the liquid chromatography conditions in accordance with the British Pharmacopoeia (BP) 2020 edition, Volume II, page 1152. (1) Impurity A: 0.13% (2) Impurity B: Not detected (3) Impurity C: Not detected (4) Impurity D: Not detected (5) Impurity E: Not detected (6) Impurity F: Not detected (7) Single unknown impurity: Not detected;

[0071] Only impurity A was detected in tiotropium bromide, accounting for 0.13% of the total impurities.

[0072] Example 3: Micronization of Salmeterol Xinafoate

[0073] Take the salmeterol xinafoate of Example 1 and use a DecJet 30 air flow mill, connect it to compressed air with a dew point temperature below -40°C, the air source pressure is greater than 12 bar, adjust the Venturi pressure to 11 bar, the rotation pressure to 10 bar, control the feed rate to 0.5 g per minute for pulverization, and collect the pulverized materials in a drying bottle for later use.

[0074] Example 4: Micronization of Tiotropium Bromide

[0075] Take the tiotropium bromide of Example 2, use a DecJet 30 air flow mill, connect it to compressed air with a dew point temperature lower than -40°C (-41.3°C on the meter), the air source pressure is greater than 12 bar, the Venturi pressure is adjusted to 11 bar, the rotation pressure is 10 bar, and the feed rate is controlled to be 0.5 g per minute for pulverization. After pulverization, collect it in a drying bottle for later use.

[0076] Example 5: Preparation of glycine carrier:

[0077] Control the operating environment humidity at 35±5%;

[0078] A Quadro U5c hammer mill was used, with the main cutter head crushing speed set to 8000 rpm, the feeding speed set to 25 rpm, and a 0.2 mm sieve installed. ≥90% of glycine (for injection) that could not pass through an 80-mesh sieve was put into the hammer mill, and glycine powder (to be screened) was obtained.

[0079] Use RUSSELL's Finex Separator grading screen to install a 90um screen, put in glycine micropowder (to be screened), screen and collect the material under the screen to obtain glycine micropowder.

[0080] Glycine powder was pulverized using a DecJet 100 air flow mill, connected to compressed air with a dew point temperature below -40°C (-42.1°C on the meter), with an air source pressure greater than 6 bar, the Venturi pressure was adjusted to 4 bar, the rotary pressure was 6 bar, and the feed rate was controlled to be 50 g per minute. The glycine ultrafine powder was collected and vacuum packed for later use.

[0081] Example 6: Lactose carrier preparation:

[0082] Control the operating environment humidity at 35±5%.

[0083] The D10 of lactose SV001 is 139; D50 is 226; and D90 is 312.

[0084] A Quadro U5c hammer mill was used, with the main cutter head crushing speed set to 8000 rpm, the feeding speed set to 25 rpm, and a 0.2 mm sieve installed. Lactose SV001 was put into the hammer mill, and the lactose SV001 powder (to be screened) was obtained.

[0085] Use RUSSELL's Finex Separator grading sieve to install a 90um sieve, put in lactose SV001 micro powder (to be sieved), sieve and collect the sieve to obtain lactose SV001 micro powder.

[0086] The lactose SV001 powder was pulverized using a DecJet 100 air flow mill, connected to compressed air with a dew point temperature below -40°C (-44.7°C on the meter), with an air source pressure greater than 6 bar. The Venturi pressure was adjusted to 4 bar, the rotary pressure was 6 bar, and the feed rate was controlled to be 50 g per minute. The lactose SV001 ultrafine powder was collected and vacuum packed for later use.

[0087] Lactose L100 is commercially available inhaled lactose, with D10 of 58, D50 of 132, and D90 of 214, and is vacuum packed.

[0088] Lactose L200 is commercially available inhaled lactose, with D10 of 9, D50 of 72, and D90 of 149, and is vacuum packed.

[0089] Lactose L201 is commercially available inhaled lactose, with D10 of 3, D50 of 22, and D90 of 59, and is vacuum-packed.

[0090] Example 7: Preparation of dry powder inhalation formulation 1

[0091] Procedure

[0092] The salmeterol xinafoate of Example 1, the tiotropium bromide of Example 3, and the glycine superfine powder of Example 5 were weighed and set aside to obtain a prescription 1, as shown in Table 1.

[0093] Table 1: Prescription 1, each inhalation 10 mg, containing 18 μg tiotropium bromide and 50 μg salmeterol xinafoate

[0094] Element Single tablet dosage Total dosage per 100,000 tablets (g) Salmeterol Xinafoate 50 μg 5 g Tiotropium bromide 18 μg 1.8 g Glycine Ultrafine Powder 9.932 mg 993.2 g total 10 mg 1000 g

[0095] Before use, salmeterol xinafoate, tiotropium bromide, and glycine ultrafine powders were manually sieved through a 100-mesh sieve in a waterless box (the humidity of the operating environment was controlled to be ≤20%) to destroy agglomerates in the package.

[0096] The humidity of the operating environment is controlled at 30±5%, 1 / 3 of the prescription weight of glycine superfine powder is placed in a high shear mixer and mixed at a low speed of 100 rpm for 30 seconds, salmeterol xinafoate, tiotropium bromide and 2 / 3 of the prescription weight of glycine superfine powder are placed in the high shear mixer together, the mixture is mixed at a high speed of 1500 rpm for 3 minutes, and then mixed at a low speed for 60 seconds to obtain the salmeterol xinafoate tiotropium bromide composition.

[0097] The humidity of the operating environment was controlled at 35±5%, and the composition was loaded into a gelatin hollow capsule (manufacturer: Suzhou Capsule Co., Ltd.; specification: 3#) by Hanhui vacuum drum filling.

[0098] The filled composition was packaged by Hualian DPH380 aluminum-plastic blister packaging machine within 6 hours, with the heat sealing temperature of 130°C.

[0099] The aluminum-plastic composite is manually simulated packaged into a finished product, and the packaging environment temperature is ≤30°C.

[0100] Example 8: Preparation of dry powder inhalation formulation 2

[0101] Procedure

[0102] The steps of Example 7 were repeated, except that a dry powder inhalation preparation was prepared using salmeterol xinafoate, tiotropium bromide, and lactose SV001 ultrafine powder according to Table 2 below.

[0103] Table 2: Prescription 2, 10 mg per inhalation, containing 18 μg tiotropium bromide and 50 μg salmeterol xinafoate

[0104] Element Single tablet dosage Total dosage per 100,000 tablets (g) Salmeterol Xinafoate 50 μg 5 g Tiotropium bromide 18 μg 1.8 g Lactose SV001 ultrafine powder 9.932 mg 993.2 g total 10 mg 1000 g

[0105] Example 9: Preparation of dry powder inhalation formulation 3

[0106] Procedure

[0107] The steps of Example 7 were repeated, except that salmeterol xinafoate, tiotropium bromide and lactose L100 were used to prepare a dry powder inhalation preparation according to Table 3 below.

[0108] Table 3: Prescription 3, 10 mg per inhalation, containing 18 μg tiotropium bromide and 50 μg salmeterol xinafoate

[0109] Element Single tablet dosage Total dosage per 100,000 tablets (g) Salmeterol Xinafoate 50 μg 5 g Tiotropium bromide 18 μg 1.8 g Lactose L100 9.932 mg 993.2 g total 10 mg 1000 g

[0110] Example 10: Preparation of dry powder inhalation formulation 4

[0111] Procedure

[0112] The steps of Example 7 were repeated, except that salmeterol xinafoate, tiotropium bromide and lactose L200 were used to prepare a dry powder inhalation preparation according to Table 4 below.

[0113] Table 4: Prescription 4, 10 mg per inhalation, containing 18 μg tiotropium bromide and 50 μg salmeterol xinafoate

[0114] Element Single tablet dosage Total dosage per 100,000 tablets (g) Salmeterol Xinafoate 50 μg 5 g Tiotropium bromide 18 μg 1.8 g Lactose L200 9.932 mg 993.2 g total 10 mg 1000 g

[0115] Example 11: Preparation of dry powder inhalation formulation 5

[0116] Procedure

[0117] The steps of Example 7 were repeated, except that salmeterol xinafoate, tiotropium bromide and lactose L201 were used to prepare a dry powder inhalation preparation according to Table 5 below.

[0118] Table 5: Prescription 5, 10 mg per inhalation, containing 18 μg tiotropium bromide and 50 μg salmeterol xinafoate

[0119] Element Single tablet dosage Total dosage per 100,000 tablets (g) Salmeterol Xinafoate 50 μg 5 g Tiotropium bromide 18 μg 1.8 g Lactose L201 9.932 mg 993.2 g total 10 mg 1000 g

[0120] Example 12: Preparation of dry powder inhalation formulation 6

[0121] Procedure

[0122] According to Table 6 below, a dry powder inhalation preparation was prepared using salmeterol xinafoate, tiotropium bromide, lactose L100 and lactose L200.

[0123] Table 6: Prescription 6, 10 mg per inhalation, containing 18 μg tiotropium bromide and 50 μg salmeterol xinafoate

[0124] Element Single tablet dosage Total dosage per 100,000 tablets (g) Salmeterol Xinafoate 50 μg 5 g Tiotropium bromide 18 μg 1.8 g Lactose L100 3.311 mg 331.1g Lactose L200 6.621 mg 662.1g total 10 mg 1000 g

[0125] Before use, salmeterol xinafoate, tiotropium bromide, lactose L100, and lactose L200 were manually sieved through a 100-mesh sieve in a waterless box (the humidity of the operating environment was controlled to be ≤20%) to destroy agglomerates in the package.

[0126] The humidity of the operating environment is controlled at 30±5%, 1 / 3 of the prescription weight of lactose L100 and 1 / 3 of the prescription weight of lactose L200 are placed in a high shear mixer and mixed at a low speed of 100 rpm for 30 seconds, salmeterol xinafoate, tiotropium bromide, 2 / 3 of the prescription weight of lactose L100, and 2 / 3 of the prescription weight of lactose L200 are placed in the high shear mixer together, the mixture is mixed at a high speed of 1500 rpm for 3 minutes, and then mixed at a low speed for 60 seconds to obtain the salmeterol xinafoate tiotropium bromide composition.

[0127] The humidity of the operating environment was controlled at 35±5%, and the composition was loaded into a gelatin hollow capsule (manufacturer: Suzhou Capsule Co., Ltd.; specification: 3#) by Hanhui vacuum drum filling.

[0128] The filled composition was packaged by Hualian DPH380 aluminum-plastic blister packaging machine within 6 hours, with the heat sealing temperature of 130°C.

[0129] The aluminum-plastic composite is manually simulated packaged into a finished product, and the packaging environment temperature is ≤30°C.

[0130] Example 13: Fine Particle Fraction of Composition

[0131] The inhalation powder aerosol test method was used according to Device 3 in 0951 of the Chinese Pharmacopoeia (2020 Edition), see Table 7.

[0132] Table 7: Fine particle fractions of tiotropium and salmeterol

[0133] prescription Tiotropium bromide (%) Salmeterol (%) Prescription 1 37.65 21.45 Prescription 2 34.83 20.96 Prescription 3 36.79 22.17 Prescription 4 32.54 19.86 Prescription 5 33.91 21.13 Prescription 6 35.37 22.29

[0134] Example 14: Delivery Dosage Uniformity of Composition

[0135] The two active ingredients, tiotropium bromide and salmeterol, were tested using the test method for uniformity of inhalation powder delivery in 0111 inhalation preparations of the "Chinese Pharmacopoeia (2020 Edition)", see Table 8.

[0136] Table 8: Tiotropium and salmeterol delivery uniformity

[0137] prescription Tiotropium bromide (%) Salmeterol (%) Prescription 1 91.7 92.1 Prescription 2 92.8 89.3 Prescription 3 89.7 91.4 Prescription 4 93.6 91.9 Prescription 5 87.9 89.6 Prescription 6 90.3 85.8

[0138] Example 15: Total impurities of the composition

[0139] The total impurities of tiotropium bromide were determined by HPLC using the liquid chromatography conditions on page 1152 of Volume II of the British Pharmacopoeia (BP) 2020 edition.

[0140] The total impurities of salmeterol were determined by HPLC using the United States Pharmacopoeia liquid chromatography conditions.

[0141] The test results are shown in Tables 9 and 10.

[0142] Table 9: Total Miscellaneous Table of Tiotropium and Salmeterol (Day 0)

[0143] prescription Total impurities of Tiotropium bromide (%) Total impurities of salmeterol (%) Prescription 1 0.12 0.06 Prescription 2 0.15 0.09 Prescription 3 0.14 0.13 Prescription 4 0.13 0.07 Prescription 5 0.14 0.11 Prescription 6 0.13 0.09

[0144] Table 10: Total Miscellaneous Table of Tiotropium and Salmeterol (90 days)

[0145] prescription Total impurities of Tiotropium bromide (%) Total impurities of salmeterol (%) Prescription 1 0.27 0.19 Prescription 2 0.29 0.24 Prescription 3 0.33 0.21 Prescription 4 0.31 0.25 Prescription 5 0.26 0.17 Prescription 6 0.35 0.23

[0146] Example 16: Stability study of the composition

[0147] The commercially available tiotropium bromide single preparation Spiriva was purchased, batch number: 107147 as contrast agent 1, and batch number: 302249 as contrast agent 2. The delivery uniformity and fine particle fraction of contrast agent 1 and contrast agent 2 were tested using the device included in the packaging box. The results of the stability study are shown in Tables 11, 12, and 13.

[0148] Table 11: Accelerated Stability Study Day 0

[0149]

[0150] Table 12: Accelerated stability study for 3 months

[0151]

[0152] Table 13: Accelerated stability study for 6 months

[0153]

[0154] Effect evaluation:

[0155] The inhalation dry powder preparation containing salmeterol and tiotropium salt of the present invention has significant pharmacodynamic advantages. Through precise synthesis and preparation processes, the preparation achieves an efficient combination of two active ingredients, significantly improving the therapeutic effect of the drug. In terms of pulmonary delivery, its fine particle characteristics ensure that the drug can penetrate deep into the lungs to achieve precise treatment. In addition, the stability study of the preparation showed that key indicators such as drug content, impurities and fine particle fractions remained stable during the 6-month observation period, indicating that the preparation of the present invention has good storage stability. Compared with commercially available contrast agents, the preparation of the present invention excels in terms of delivery dosing uniformity and fine particle fraction, providing patients with a more reliable and effective treatment option.

Claims

1. A dry powder preparation for inhalation containing salmeterol or a pharmaceutically acceptable salt thereof and tiotropium salt, characterized in that: Comprising salmeterol or a pharmaceutically acceptable salt thereof, tiotropium salt and at least one pharmaceutically acceptable carrier selected from lactose, sucrose, mannitol and amino acid; The salmeterol or its pharmaceutically acceptable salt is micronized and mixed with tiotropium salt and the pharmaceutically acceptable carrier; Wherein, the weight ratio of the tiotropium salt: salmeterol or its pharmaceutically acceptable salt: pharmaceutically acceptable carrier is 1:1-15:500-50000, preferably 1:3:550; The synthesis of salmeterol comprises the following steps: 2-hydroxybenzaldehyde and bromoacetyl bromide undergo Friedel-Crafts acylation to generate an intermediate (I), 4-phenyl-1-butanol and 1,6-dibromohexane undergo reaction to generate an intermediate (II), separation, crystallization and extraction to obtain an intermediate (III), and intermediates (I) and (III) undergo condensation followed by reduction catalytic hydrogenation and debenzylation to obtain salmeterol, but the synthesis is not limited to this route.

2. The dry powder preparation for inhalation according to claim 1 or 2, characterized in that The pharmaceutically acceptable carrier is a pharmaceutically acceptable carrier selected from lactose, sucrose, mannitol, and amino acids, and glycine is particularly preferred.

3. A method for preparing the dry powder formulation for inhalation according to any one of claims 1 to 3, the method comprising the following steps: Step 1) synthesizing salmeterol or a pharmaceutically acceptable salt thereof; Step 2) synthesizing tiotropium salt; Step 3) micronizing salmeterol or a pharmaceutically acceptable salt thereof and tiotropium salt respectively; Step 4) weighing micronized salmeterol or a pharmaceutically acceptable salt thereof, tiotropium salt and a pharmaceutically acceptable carrier according to a predetermined ratio and mixing them uniformly; Step 5) The mixture is filled into capsules and packaged to produce a commercial product.

4. The method according to claim 4, characterized in that: In step 4, mixing is performed by shear mixing.

5. The method according to claim 4 or 5, characterized in that: During the micronization process, the humidity of the operating environment is controlled at 25% to 45%, preferably 30%.

6. The method according to any one of claims 4 to 6, characterized in that: The capsule content composition is administered by oral inhalation using an inhalation device and delivered to the lungs.

7. The method according to claim 7, characterized in that: The inhalation device is a powder mist inhalation device disclosed in CN219423486U by Nanchang Hongyi Pharmaceutical Co., Ltd.

8. Use of the dry powder preparation according to claims 1 to 2 in the preparation of a medicament for treating respiratory diseases such as asthma and chronic obstructive pulmonary disease (COPD).

9. The dry powder preparation according to any one of claims 1 to 2, wherein the salmeterol or a pharmaceutically acceptable salt thereof is preferably salmeterol xinafoate, and the tiotropium salt is preferably tiotropium bromide.

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