WO2017124640A1 - Alendronate sodium powder inhalation used for respiratory drug deliver and application thereof - Google Patents

Abstract

Alendronate sodium powder inhalation used for respiratory drug delivery and application thereof. The alendronate sodium powder inhalation contains single-component alendronate sodium micro powder, without containing auxiliaries. The alendronate sodium powder inhalation can improve the breathing function of a rat with COPD by about 40%, better than the effect of salbutamol sulfate in a conventional dosage, and can overcome the drug resistance problem of salbutamol sulfate and other beta2 receptors.

Description

Alendronate sodium powder for respiratory administration and use thereof

The invention belongs to the field of pharmacy and preparation, and relates to an alendronate sodium powder for respiratory administration, a preparation method thereof and a use thereof, in particular, alen which mainly exerts pharmacodynamic effects after being inhaled mainly in the respiratory tract after inhalation. Sodium phosphonate powder.

Sodium Alendronate is a small molecule compound with a molecular weight of 249.97 and is structurally similar to the sodium aminophosphate pyrophosphate. The existing dosage form is oral for tablets, and the main effect is to inhibit bone resorption, and is used for treating and preventing osteoporosis in postmenopausal women. The pharmacological mechanism of its action is based on the affinity of alendronate and bone hydroxyapatite crystals to selectively inhibit osteoclast resorption in the bone reforming cycle. In 1995, the US FDA approved alendronate tablets for oral treatment of Paget's disease and postmenopausal osteoporosis in women. The clinical dose is about 0.2 mg/kg (10 mg/day) orally. Oral alendronate tablets form a stimulating alendronate under the action of gastric acid, which may cause gastrointestinal discomfort, may also cause stomach and esophageal damage and ulcers; even lead to esophageal cancer, therefore, presenting the gastrointestinal Patients with tract problems or esophageal disorders such as sputum sputum should not take alendronate tablets. High blood entry into alendronate, such as intravenous alendronate, was observed in rats and poses a risk of kidney toxicity and leads to accumulation of alendronate in non-calcified tissue. The local absorption of alendronate is better: subcutaneous and intramuscular injection can be performed.

Chronic obstructive pulmonary disease ("slow obstructive pulmonary disease" or "COPD") is a chronic lung disease characterized by incomplete reversible airflow limitation. It is often clinically characterized by recurrent cough, cough, and difficulty breathing. Symptoms, usually characterized by progressive progression, include most chronic bronchitis and emphysema. As the disease progresses, the airway remodels, eventually developing into irreversible airflow obstruction, or coexisting with asthma. Asthma (full name: bronchial asthma) is a chronic respiratory disease, usually caused by airway inflammation, divided into acute bronchoconstriction, leading to repeated attacks of wheezing, chest tightness, difficulty breathing, and cough. COPD and asthma pose a serious threat to public health on a global scale. There are currently 210 million COPD patients and approximately 300 million asthma patients worldwide.

2-2-receptor agonists (including salbutamol, salmeterol, salmeterol, etc.) are currently the most effective bronchodilators, which can quickly relieve bronchospasm and clinical symptoms, and are the treatment of choice for all ages of COPD and asthma. However, long-term (1 to 2 weeks or longer) use of a β2 receptor agonist can lead to a decrease in drug tolerance and even loss of efficacy. The clinical use of β2 receptor agonists has been limited due to the problem of tolerance, and is currently mainly used as a relief drug, either as needed, or in combination with glucocorticoids. Glucocorticoids are currently the most commonly used control drugs (ie, drugs used every day), but glucocorticoids do not cure asthma and require long-term use, which can cause many adverse reactions to patients.

Japanese Sasaki et al. and Ueno et al. respectively verified that alendronate has anti-inflammatory effects on asthma and COPD mice by mouse experiments. However, these effects are directly added to the culture medium by cell culture or oral administration, and there is no test or direct administration through the respiratory tract. Because the characteristics of alendronate sodium after blood entry are quickly excreted mostly through the kidneys, a small part of it is deposited in the bones, so that the efficiency of inhaling into the bloodstream through absorption into the blood is very low, and the side effects such as kidney toxicity are increased. The risk of such small particle size alendronate micropowder and its powder is not ideal for COPD or asthma medication.

In order to overcome at least one of the deficiencies of the prior art, the present invention provides a preparation of an alendronate sodium powder spray, which can improve the local utilization rate in the respiratory tract while reducing intra-alveolar inhalation to reduce blood volume and avoid causing Side effects on the body system.

The present invention provides an alendronate sodium powder for respiratory administration, wherein the alendronate sodium powder has a single alendronate sodium micropowder and contains no excipients.

Further, the alendronate sodium powder is prepared by directly pulverizing the alendronate sodium raw material.

Preferably, the present invention contemplates the preparation and use of a larger particle size of alendronate sodium powder, which has not been reported in the prior literature. Among the dosage forms for pulmonary inhalation administration, the powder loading amount of the powder is large, and the production process is relatively simple. According to the design and application of the powder, the particle size distribution, solubility, bulk density, angle of repose, atomization performance and moisture absorption of the powder must be considered in order to achieve curative effect. The conventionally used powder is intended to enter the alveoli and then enter the bloodstream, and the powder from the mouth to the lungs passes through a relatively long moist respiratory tract such as the trachea and bronchi, while particles with a particle size of 5 μm or more are easily deposited in the respiratory tract. Medium and can not enter the alveoli, so the particle size (d90) generally needs to be controlled below 5μm, and often need to use water-soluble, irritating, biocompatible excipients to facilitate drug diffusion into the alveoli, reduce and avoid in the respiratory tract The deposition in the middle. These excipients, such as surfactants, may cause damage to the lungs when used for a long time. We consider that the main symptoms of COPD and asthma are tracheal and bronchial obstruction. The powder with moderate particle size (5~20μm) can be directly deposited in the trachea, bronchi and bronchioles, which is equivalent to local application in the respiratory tract. It works faster and avoids or mitigates possible system side effects. According to the characteristics of granules in the respiratory tract: 5~20μm mainly deposited in the trachea, bronchus and bronchioles, ≤2.5μm can enter the small bronchi and in the alveolar deposition, the size of the aerosol applied to the trachea, bronchi and bronchioles (d90) Suitable for control in the range of 5~20μm.

Preferably, the fine powder particle size distribution of the alendronate sodium powder is d90≤20 μm, and d50≥5 μm, d10≤2.5 μm.

Further, the alendronate sodium micropowder is filled into a capsule with a charge of 0.6 mg to 5 mg, and is administered once a day.

The invention also provides a preparation method of the above-mentioned alendronate sodium powder aerosol, which is prepared by directly pulverizing alendronate sodium raw material medicine without adding an auxiliary material or a solvent.

Further, the pulverization loading amount is 50 g to 1000 g, the pulverization pressure is 0.7 MPa, the pulverization time is 2.5 h to 3.5 h, and the pulverization times are 1 time or more.

The present invention also provides the use of the above alendronate sodium powder for the preparation of a medicament for treating chronic obstructive pulmonary disease and asthma.

Further, the present invention provides the use of the above-described alendronate sodium powder for the treatment of chronic obstructive pulmonary disease in a patient resistant to a β2 receptor agonist.

The alendronate sodium powder of the invention is prepared by directly pulverizing the main drug of alendronate, and does not need to use or add any auxiliary materials. Compared with the spray drying method reported in the literature, the method not only produces the method. The preparation process is simple, easy to implement, and avoids the side effects that may be caused by the use of excipients and solvents in the spray drying process.

The alendronate sodium powder of the present invention mainly deposits in the respiratory tract to directly exert the bronchodilator effect. Due to the larger particle size, the portion that enters the alveoli is smaller, so the amount of blood entering is low, and deposition into other tissues and kidney pressure is avoided.

The alendronate sodium powder of the present invention can not only enhance the efficacy of the β2 receptor agonist, overcome the problem of tolerance, and can reduce the amount of β2 receptor agonist used, reduce and avoid β2 Side effects of receptor agonists (mainly in the heart). In addition, alendronate has an anti-inflammatory effect, the amount of glucocorticoids can be reduced accordingly, and the side effects of glucocorticoids can also be alleviated. Meanwhile, the alendronate sodium powder of the present invention contains a small amount of a component having a small particle size and capable of entering the alveolar absorption, and this part of the alendronate which is absorbed into the blood may exert anti-inflammatory and prevent osteoporosis. To help prevent and overcome the side effects of inhaled glucocorticoids, therefore, the alendronate sodium powder of the present invention only needs to control d90 ≤ 20 μm, and d50 ≥ 5 μm, d10 ≤ 2.5 μm, and it is not necessary to exclude the particle size at 2.5 μm. The following sections. Therefore, the alendronate sodium powder of the present invention is expected to develop into a therapeutic drug for COPD and asthma, and is suitable for long-term use in patients with COPD and asthma.

Figure 1 is a SEM electron micrograph of an ALE-4 sample.

Figure 2 is a SEM electron micrograph of an ALE-6 sample.

Figure 3 is a high performance liquid chromatogram of alendronate sodium micropowder.

Figure 4 is a H&E staining diagram of rat lung tissue.

Figure 5 is a graph showing the relative parameter changes of specific airway resistance after inhalation of a gradient concentration of salbutamol in each rat.

Figure 6 is a graph showing the relative parameter changes of specific airway resistance after inhalation of a gradient concentration of alendronate in each rat.

Figure 7 is a graph showing the reactivity of four normal rats to salbutamol on day 0.

Figure 8 is a graph showing the reactivity of four COPD rats to salbutamol on day 0.

Figure 9 is a graph showing the mean response to salbutamol on Day 0 in four normal and four COPD rats.

Figure 10 is a graph showing the reactivity of salbutamol in four normal groups of rats after 21 days of salbutamol tolerance induction.

Figure 11 is a graph showing the reactivity of salbutamol in four COPD rats after 21 days of salbutamol tolerance induction.

Figure 12 is a graph comparing the mean values of salbutamol reactivity in 4 normal groups and 4 COPD rats after 21 days of salbutamol tolerance induction.

Figure 13 is a graph comparing the mean values of salbutamol reactivity in 4 COPD rats before and after salbutamol tolerance induction.

Figure 14 is a graph showing the reactivity of four normal rats to alendronate after 21 days of salbutamol tolerance induction.

Figure 15 is a graph of response to alendronate in 4 COPD rats after 21 days of salbutamol tolerance induction.

Figure 16 is a graph showing the mean response to alendronate in four COPD rats after 21 days of salbutamol tolerance induction.

Figure 17 is a graph comparing the mean values of reactivity of albendronate to four normal rats and four COPD rats before and after salbutamol tolerance induction.

The above and other objects, features, and advantages of the present invention will become more apparent and understood by the appended claims appended claims

An embodiment of the present invention provides an alendronate sodium powder for respiratory administration, which comprises a single alendronate sodium micropowder and contains no excipients.

It should be understood that the above meaning without the auxiliary material means that no other ingredients are added in the preparation of the alendronate sodium powder of the present invention, and the impurities contained in the preparation of the alendronate sodium raw material are not considered to be Excipients.

Preferably, the purity of alendronate sodium in the alendronate sodium powder or the alendronate sodium drug substance used in the embodiment of the invention is ≥98%.

According to a preferred embodiment of the invention, the particle size distribution of the alendronate sodium aerosol satisfies the following conditions: d90 ≤ 20 μm, d50 ≥ 5 μm, and d10 ≤ 2.5 μm. In the present specification, d90, d50 and d10 represent a particle size distribution. For example, the meaning of d90 is: the particle diameter corresponding to a cumulative particle size distribution of a sample of 90%, and its physical meaning is that the particle diameter is smaller than the value (20 μm). The particles account for 90%.

According to one embodiment of the invention, the alendronate sodium powder is prepared by directly pulverizing the alendronate sodium drug substance. According to another embodiment, the alendronate sodium powder is prepared directly during the preparation of the alendronate sodium bulk drug, that is, the particle size distribution of the powder is obtained directly without a tableting operation.

Embodiments of the present invention also provide the use of the alendronate sodium powder of the present invention for the treatment of chronic obstructive pulmonary disease and asthma, and chronic obstructiveness of a patient for resistance to a β2 receptor agonist The use of lung disease.

According to a preferred embodiment, for the treatment of chronic obstructive pulmonary disease and asthma, the dose is from 0.6 mg to 5 mg per day, and is administered in the form of a capsule.

Formula

The invention adopts alendronate sodium raw material medicine which meets the national standard as a raw material, is prepared by mechanical pulverization method, and is not used or involved in other auxiliary materials in the production and preparation process of the fine powder.

1. Micro powder preparation process. The present invention employs a mechanical pulverization method.

Mechanical pulverization is one of the common methods for preparing powders, but it has gradually been replaced by spray drying. The main reason is that compared with the spray drying method, the fine powder prepared by the mechanical pulverization method has a larger particle size, 2 to 3 μm is the pulverization limit of the method, and the shape of the fine powder is irregular; the pulverization can also generate static electricity, and after the micronization of the drug, The higher surface free energy, the particles tend to aggregate, resulting in enhanced viscosity of the drug micropowder, poor fluidity, affecting the redispersion of the drug after release from the micropowder inhaler. The severe pulverization of the mechanical pulverization process may also cause adverse physicochemical properties of the drug. In particular, amorphous crystals may be formed on the surface of the fractured crystal. However, the inventor's research suggests that for the local administration of the respiratory tract, the mechanical pulverization method is advantageous in that the fine powder has a large particle size and irregular shape, which is favorable for local deposition in the respiratory tract; the preparation process is simple, the production cycle is short, and the chance of microbial contamination is very high. Low; since it is not necessary to use an auxiliary material such as a solvent, as long as the pulverization pressure and time are properly controlled, the main drug is not easily deteriorated. We consider that this powder is designed for local administration of the respiratory tract. It does not require strong fluidity. The fine particle size is larger and more favorable, and alendronate is a small molecule drug with simple structure, and its physical and chemical properties are relatively stable. It can withstand the processing of mechanical pulverization. Therefore, we choose mechanical pulverization. The pulverization conditions mainly refer to the operating conditions of the tiotropium bromide inhalation powder, and two kinds of pulverizers such as QS100 and AO type were used for the test. The pulverization pressure is controlled at 0.7 MPa, and the single continuous pulverization time does not exceed 3.5 hours. Under the pulverization conditions, alendronate did not deteriorate. The fine powder particle size was measured using a Mastersizer 2000 laser particle size analyzer, and the results are shown in Table 1 below.

Sample serial number	Crushing equipment	Loading amount (g)	Smashing time (h)	Crushing pressure (Mpa)	Number of shredding	D9 (μm)	D50 (μm)	D10 (μm)
ALE-1	QS100	1000	3.5	0.7	1	37.36	10.57	2.43
ALE-2	QS100	1000	3.0	0.7	2	25.53	8.77	2.47
ALE-3	QS100	1000	3.5	0.7	3	19.59	9.66	2.22
ALE-4	QS100	1000	3.5	0.7	4	19.58	7.78	2.14
ALE-5	AO	50	2.5	0.7	1	12.32	5.65	1.75
ALE-6	AO	50	2.5	0.7	2	11.43	5.25	1.72

It can be seen from the above Table 1 that the particle size distribution is related to the amount of the sample and the number of pulverization under the pulverization conditions in which the pulverization pressure is controlled at 0.7 MPa and the single continuous pulverization time is not more than 3.5 hours. The loading amount of 1000 g and pulverization 3 times or more (3.0 to 3.5 h) can control the particle size D90 to be less than 20 nm (19.58 to 19.59 μm). Among them, 4 times of pulverization and 3 times of pulverization have little particle size reduction, indicating that under the condition of 1000 g of sample loading, the pulverization effect of 3 times and 3 times or more is basically equivalent, so the number of pulverization of 1000 g can be selected 3 times. When the loading amount is 50 g and the pulverization is performed once or more (2.5 h), the particle size D90 can be controlled to about 12 μm (11.43 to 12.32 μm). Among them, the pulverization and the pulverization particle size reduction were small, indicating that the pulverization effect of one time and one time is basically equivalent under the condition of 1000 g of the sample loading amount, so the number of pulverization of 50 g can be selected once. According to the literature, the particle size of 7~20nm belongs to the range of the respiratory tract (trachea and bronchus). Therefore, these two conditions (1000g sample loading smash 3 times or more, 50g sample loading smash 1 time or more) are consistent. Our requirement. We took ALE-4 and ALE-6 samples as representative samples of the two preparation conditions, respectively, and performed SEM electron microscopy analysis, as shown in Fig. 1 and Fig. 2. 1 is a SEM electron micrograph of an ALE-4 sample, and FIG. 2 is a SEM electron micrograph of an ALE-6 sample.

We performed two methods for ALE-4 and ALE-6, such as solubility, ninhydrin identification, HPLC identification, molybdenum blue colorimetric determination, microbial contamination and various physical and chemical properties.

2.1 Solubility: 10 mg of each of the experimentally prepared ALE-4 and ALE-6 alendronate sodium powders were weighed and placed in a stoppered tube containing 2 ml of water, and shaken for 1 minute to observe the dissolution. Both powders were completely soluble and the solution was clear, indicating that the solubility was acceptable.

Characterization: 10mg of each of ALE-4 and ALE-6, dissolved in 2ml of water, then add 1ml of ninhydrin test solution, mixed, heated and boiled for 10 minutes, all purple. The absorbance was measured at 568 nm, and the absorbance was very close to the drug substance.

2.2 HPLC identification:

Alendronate sodium was determined by pre-column derivatization. The molecular structure of alendronate is relatively simple and lacks corresponding chromophores. The amino group in the alendronate molecule is combined with the FMOC reaction to form a fluorescent group.

High-pressure liquid chromatography and its model: Shimadzu Shimadzu LCsolution

Chromatographic conditions:

Column: Shiseido Capcell Pak C18 (150 mm × 4.6 mm, 5 μm, Japan Shiseido Co., Ltd.).

Mobile phase:

(1) Solution A: acetonitrile; Solution B: 0.01 M disodium hydrogen phosphate, gradient elution procedure: 0 to 24                 Min, 86% A; 24 to 29 min, 30% A; 29 to 30 min, 86% A; column temperature: 35 ° C;                 Flow rate: 1.0 mL•min-1.

(2) Solution A: acetonitrile; solution B: 25 mM citric acid, 25 mM pyrophosphate. Gradient elution program: 0~24                 Min, 86% A; 24 to 29 min, 30% A; 29 to 30 min, 86% A; column temperature: 35 ° C;                 Flow rate: 1.0 mL•min-1.

Loading volume: 1 uL or 2 uL.

Fluorescence detector:

Fluorescence excitation wavelength: 260 nm, fluorescence emission wavelength: 310 nm.

The chromatogram was obtained by eluting with a gradient of 25 mM pyrophosphate of acetonitrile-25 mM citric acid as shown in FIG. Among them 3.5min -                 The alendronate peak was at 4 min and the Fmoc peak at 13 min-14.5 min. Both ALE-4 and ALE-6 showed alanine sodium peak, and the peak height was comparable to that of the drug substance, indicating that there was no main drug degradation of ALE-4 and ALE-6.

2.3 Determination of molybdenum blue colorimetric method:

Take 8mg of ALE-4 and ALE-6, dissolve in water, add water to the 100ml volumetric flask, heat in a 40°C water bath for 20 minutes, shake constantly, let cool to room temperature, dilute with water to the mark, serve as The test solution; another accurately weighed 10 mg of alendronate sodium raw material, placed in a 100 ml volumetric flask, dissolved in water, diluted with water to the mark, shaken, as a reference solution. Precisely measure 5ml of the test solution and the reference solution, place them in a 25ml volumetric flask, add 8ml of ammonium persulfate solution (1%), heat in a water bath for 20 minutes, let cool to room temperature, add ammonium molybdate solution (take 7.5g of ammonium molybdate, dissolve in 100ml of water, add 100ml of 5mol/L sulfuric acid solution, mix well) 2.0ml, shake well, leave for 15 minutes, add p-aminophenol sulfate solution (take p-aminophenol sulfate 0.5g, add 195ml of 15% sodium bisulfite solution, add 5ml of 20% sodium sulfite solution, shake well 2ml, shake, place for 15 minutes, add 5ml of 34% sodium acetate solution, add water to the mark and shake. The absorbance was measured at a wavelength of 710 nm, and the absorbance of the two test solutions was comparable to that of the reference solution, indicating that there was no main drug degradation of ALE-4 and ALE-6.

2.4 Microbiological examination:

3 mg of each of ALE-4 and ALE-6 were separately taken and dissolved in 10 ml of purified water. According to the Pharmacopoeia                 In 2010, the second appendix XI J microbial limit test was carried out. The number of bacterial colonies was ≤ 2 / ml; the number of mold and yeast colonies = 0 / ml. All are qualified.

2.5 Bulk density, angle of repose, porosity, critical relative humidity:

Bulk density, porosity: The bulk density is the density obtained by dividing the mass of the powder by the volume V of the container occupied by the powder. When filling the powder, the density measured after a certain regular vibration or tapping is called the tap density ρtap . Porosity refers to the ratio of the volume of voids formed in the matrix to the total volume. After the micronization treatment of the drug, the bulk density and porosity of the drug change greatly, which may cause the density difference between the drug and the auxiliary material, which causes difficulty in mixing uniformity. Therefore, micronized drugs should be tested for bulk density and porosity. The volume measured by filling the powder into the container includes the true volume of the powder, the voids in the particles, the voids between the particles, and the like. Therefore, the shape and size of the measuring container, the filling speed of the material, and the filling method affect the volume of the powder. When the powder is loaded into the measuring container, the density measured is the loosest bulk density without applying any external force, and the density measured by applying an external force to make the powder in the tightest filling state is called the tightest packing density. The tap density changes with the number of tapping times, and the tap density measured when the final oscillation volume is constant is the tightest packing density.

Fill a certain amount of powder sample into the modified 5ml measuring cylinder, read the initial volume V, tap and observe the volume change of the powder in the measuring cylinder until the volume does not change significantly (usually tap 2500 times), record The final volume V is final. The mass difference before and after the loading is the mass w of the added powder.

Bulk density pb=w/V initial, tap density ptap=w/V final, porosity=1-stack density/tear density

Angle of repose: The angle of repose is measured by taking a small funnel with a diameter of about 6 cm and a diameter of about 0.4 cm. It is fixed on the iron frame, and a white paper is placed under the funnel. The lower end of the funnel and the height of the paper are 4-5cm, slowly pour the powder from the top of the funnel into the funnel. When the powder to be leaked is close to the funnel outlet, measure the height of the cone and the diameter of the lower end of the powder, calculate the tangent of the angle of repose, and further calculate the angle of repose. .

Determination of critical relative humidity. After the micronization treatment of the drug, the hygroscopicity may change significantly due to the increase of the specific surface area, and the moisture is a strictly controlled inspection item of the powder, so the critical relative humidity of the micronized drug should be determined (Critical)                 Relative Humidity, CRH).

The critical relative humidity (CRH) determination method is divided into two steps:

First, make a moisture absorption curve:

Weigh 2mg of the drug, spread it in a flat weighing bottle that is dry to constant weight (about 2mm thick), dry to constant weight, accurately weigh, open the cap, and hold the concentrated sulfuric acid solution or other salt solution (relative humidity) 75%) of the glass dryer, stored in an incubator at 25 ° C, take out the weighing bottle at 2, 4, 8, 12, 24, ... or other time periods, accurately weighed, calculate the percentage of moisture absorption, to absorb moisture The rate versus time plots the hygroscopic curve and finds the corresponding time T days (or hours) for the moisture absorption balance.

Second, the determination of CRH:

Weigh the drug (powder or granule) 1-2mg 6 parts, the same operation, set the glass drier with different concentrations of concentrated sulfuric acid solution or other salt solution, store in the incubator at 25 degrees Celsius for T days, take out the scale The measuring bottle is accurately weighed, the percentage of moisture absorption is calculated, and the moisture absorption balance curve is plotted against the relative humidity. The two tangents are made at the inflection point, and the relative humidity corresponding to the tangent point is CRH.

After experimental determination, the various properties of the alendronate sodium micropowder prepared by the two process conditions are not much different. Therefore, it is expected that the design goals will be met.

3. Alendronate powder spray capsule

The alendronate sodium powder of the present invention can be applied in the form of a capsule. The size of the capsule used may be any type, and the preferred model number is less than 0, usually 1-4. Alendronate capsules are not only easy to industrialize, but also easy to carry. The amount of alendronate sodium powder in the capsule is constant, ensuring accurate inhalation dose during use, and effectively preventing the patient from taking more or less suction. The micropowder is filled into a capsule in a single dose of 0.6 mg to 5.0 mg, and sealed and stored. Use in a single capsule when using. Good anti-hygroscopic effect. Satisfactory emptying and atomization performance can still be achieved in an environment where the relative humidity reaches 90%. The atomization effect and the residual amount after use produce a good synergistic effect, in particular, the hygroscopicity of the alendronate sodium powder spray can be reduced.

3.1 Determination of the uniformity of the content of the capsule. Method: sampling 30 capsules, weighing the balance

3.2 Determination of the emptying rate of the capsule. Methods: According to the 2010 edition of the Chinese Pharmacopoeia (Part 2) Appendix IL method. Emptying rate requirement ≥90%

Capsule stability, mainly concerned with the effect of water absorption of capsule shell on the performance of the preparation

Method: --- Accelerated test: Take 3 batches of capsules (30/batch) and store them in a constant temperature and humidity chamber (40±2°C, 75±5% relative humidity) at the end of 0, 1, 2 and 3 months respectively. Sampling, inspection of appearance, emptying rate, deposition rate, content

3.3 Powder residual amount and atomization performance: The prototype alendronate sodium inhalation powder is accurately weighed (W1), placed in a special inhaler (manufactured by Shanghai Tianping Pharmaceutical Factory), the capsule is perforated, and then inhaled. The unit is connected to a 5000ml glass bottle with a switch knob that is in the off position. The vacuum was applied at a flow rate of 60 l/min, and the above knob was turned on, and the powder in the capsule was ejected from the inhaler three times in succession. If the powder forms a uniform smoke, no large particles are present after deposition, indicating that the atomization performance is excellent; if most of the powder is atomized, there is only a small amount of particles at the bottom of the bottle, and the atomization performance is moderate. If most of the powder is not atomized, it is deposited in the form of a block at the bottom of the bottle, indicating poor atomization performance. The used capsule shell was taken out from the inhalation device, accurately weighed (W2), and the residual powder on the inner wall of the capsule was wiped off with a small brush, and the weight of the capsule (W3) was weighed. The calculation method of the residual amount of the capsule contents is: [(W2-W3)/(W1-W3)]×100%

Humidity effect and capsule wall adhesion test: Take alendronate sodium inhalation powder, accurately weighed, set the relative humidity of 75% at room temperature at 25 ° C, take it out after 24 hours, and weigh the capsule again. The powder was then poured out, the change in powder properties was observed, and the residual amount of the powder in the capsule was measured by the above method to understand the adhesion of the capsule wall.

Table 2 below shows the experimental results for screening alendronate sodium aerosol formulations:

	Prescription 1	Prescription 2	Prescription 3	Prescription 4	Prescription 5	Prescription 6	Prescription 7	Prescription 8
Alendronate micropowder ALE-4	5mg	2.5mg	1.2mg	0.6mg	-	-	-	-
Alendronate micropowder ALE-6	-	-	-	-	5mg	2.5mg	1.2mg	0.6mg
Solubility	qualified	qualified	qualified	qualified	qualified	qualified	qualified	qualified
Bulk density	0.28	0.28	0.28	0.28	0.32	0.32	0.32	0.32
Angle of repose	42	42	42	42	37	37	37	37
Emptying rate	91.3%	93.7%	94.6%	95.2%	92.5%	93.8%	94.7%	95.4%
Shell powder residue	7.2%	5.1%	3.9%	3.5%	6.6%	5.0%	3.7%	3.6%
Changes in capsules at 75% relative humidity	+2.1%	+2.7%	+2.5%	+2.3%	+2.0%	+2.7%	+2.9%	+2.6%
Atomization performance	medium	medium	medium	medium	medium	medium	medium	medium
Primary distribution bottle deposition rate	62.3%	61.5%	60.8%	60.2%	58.6%	58.2%	57.5%	57.7%
Secondary distribution bottle deposition rate	20.1%	20.6%	21.7%	21.5%	22.8%	22.3%	23.7%	23.5%

The above capsules composed of ALE-4, ALE-6 or ALE-4 mixed with ALE-6 have little difference in properties.

The contents of the capsules provided by the present invention consist of a fine powder of alendronate sodium having a particle size of less than 30 μm. More preferably, the fine powder has a particle size of less than 20 μm, and the most preferred fine powder has a particle size controlled at d90 ≤ 20 μm, and d50 ≥ 5 μm, d10 ≤ 2.5 μm. The solubility, bulk density, angle of repose and emptying rate of the micropowder are good, the atomization performance is medium, and it is not easy to absorb moisture during processing, storage and use. Generally, the powder for treating COPD and asthma can be inhaled to the center of the airway to produce a predetermined therapeutic effect. Since the alendronate sodium powder prepared according to the present invention has a good atomization effect, a commercially available inhalation powder device for treating COPD and asthma can be utilized. For example, use an inhaler from Shanghai Tianping Pharmaceutical Factory. The device actually functions to crush the capsule. The device for squeezing the capsule can be used repeatedly. The contents of one capsule can be discarded after the light is absorbed. The next time the product is used, a capsule is filled into the device. The alendronate sodium capsule of the present invention can also be opened by hand, and the fine powder can function by entering the respiratory tract by the action of airflow during breathing. This facilitates self-administration by patients in outdoor or emergency situations.

4.HPLC analysis of the distribution and metabolism of alendronate in lung tissue of alendronate sodium aerosol after respiratory administration

4.1 Operation steps

4.1.1 Tissue sample processing Experimental steps:

Weigh 0.5 g of lung tissue, add 300 μL of the standard solution, and grind.

Add 1 ml of 6% TCA (pH = 1), centrifuge at 4000 rpm for 10 minutes, and pour out the supernatant.

Add 0.1 mol/L potassium dihydrogen phosphate solution 200uL, 0.1                 200 uL of mol/L calcium chloride solution, 400 uL of 1 mol/L sodium hydroxide solution.

The supernatant was removed by 3000 rpm for 5 minutes.

The resulting precipitate was dissolved in 200 uL of acetic acid solution (0.2 mol/L), diluted with 0.6 mL of water, and precipitated with a 1 mol/L sodium hydroxide 50 uL solution.

The above sodium hydroxide precipitation reaction was repeated once.

The resulting precipitate was dissolved in 1 mL of a 0.2 mol/L sodium acetate buffer solution (pH 4.5).

4.1.2 Bond Elut-DEA Solid Phase Extraction Column Purification and Derivatization Labeling

Bond Elut-DEA (Company Agilent, Cat. No. 12102016), bed volume: 300 uL

The solid phase extraction column was activated with 1 ml of methanol, and the solid phase extraction column was washed twice with 1 ml of pure water.

Take 0.5 mL of sample solution through the column. The effluent was sampled at 10 uL.

Wash once with 1 ml of pure water. The effluent was sampled at 10 uL.

Wash once with 1 ml of pure water.

Elute with 1 mL of sodium pyrophosphate in the eluent. Take 300ul directly to adjust the pH for the derivatization reaction.

Take 300uL each, take 300ul of eluent directly, add 1mol/L sodium carbonate buffer solution (pH 11.9)                 100uL and FMOC working solution 50uL, shake quickly, 5min.

Add 1 mol / L citric acid 100 uL to adjust the pH value of about 6.5, (optional: solution centrifugation, 12 000                 r/min *3 min), the supernatant was taken for injection analysis at 450 uL.

The sample was loaded by HPLC, and the amount of the sample was 4 uL.

4.1.3 HPLC detection conditions

Chromatographic conditions

Column: Shiseido Capcell Pak C18 (150 mm × 4.6 mm, 5 μm, Japan Shiseido Co., Ltd.);

Mobile phase: A: acetonitrile B: 25 mM sodium pyrophosphate, 25 mM citric acid (pH 5.05)

Elution procedure: 0 to 22 min, 70% B

Column temperature: 35 ° C; flow rate: 1.0 mL•min ^-1 ;

Fluorescence excitation wavelength: 260 nm, fluorescence emission wavelength: 310 nm.

Load amount: 4uL

4.2 Distribution of alendronate sodium powder in rat lung after respiratory administration

Two SD rats were taken, and the age was 8 months. After anesthesia with isoflurane anesthesia machine, 0.32 mg of alendronate sodium was administered by a rat lung powder mist applicator. After 5 minutes, the rats were sacrificed and the lung tissues were taken out to separate the left lung: upper and lower lung lobe Right lung: upper, middle and lower lobes were weighed and ground for tissue grinding. The content of alendronate sodium in the slurry was determined by HPLC as shown in Table 3.

The first spray is calculated as 0.32 mg
Rat	Lung leaf	Weight (g)	Grinding volume (mL)	ALN concentration (ug/mL)	Total amount (mg)	% of total drug intake
	Left one	0.196	0.5	52.6	0.026	8.10%
	Left two	0.266	0.5	134.5	0.067	20.90%
No.1	Right one	0.322	0.5	165.2	0.083	25.90%
	Right two	0.283	0.5	118.8	0.059	18.40%
	Right three	0.214	0.5	63.4	0.032	10.00%
	total	1.281			0.267	83.40%
	Left one	0.232	0.5	93.3	0.047	14.70%
	Left two	0.274	0.5	113.6	0.057	17.80%
No.2	Right one	0.314	0.5	134.4	0.067	20.90%
	Right two	0.343	0.5	145.4	0.073	22.80%
	Right three	0.276	0.5	102.5	0.051	15.90%
	total	1.439			0.295	92.20%
	Left one	0.214	0.5	72.95	0.036	11.30%
	Left two	0.27	0.5	124.05	0.062	19.40%
average	Right one	0.318	0.5	149.8	0.075	23.40%
	Right two	0.313	0.5	132.1	0.066	20.60%
	Right three	0.245	0.5	82.95	0.041	12.80%
	total	1.36			0.28	87.50%

the result shows:                 After the tracheal administration of the alendronate powder, it can be distributed in each lung lobe, especially in the lobe near the main bronchus. About 5 minutes, the total dose of the lung leaves accounted for 87.5% of the input dose. It indicates that the alendronate sodium powder has good air dispersion characteristics in terms of particle size and dispersion, and is easy to be uniformly absorbed by the pulmonary administration.

HPLC chromatogram: The peak at 3.4-3.9 min is the peak of alendronate sodium.

5. Alendronate sodium aerosol (ALN) non-clinical pharmacodynamics study (pharmacodynamic animal experiment)

experiment material

(1) Animals: 72 SD rats were clean, of which 36 were females with an average body weight of 176±22 g; and 36 males each with an average body weight of 195±25 g. Provided by the Experimental Animal Center of Shanghai Pulmonary Hospital.

(2) Drugs: 1) Alendronate sodium powder. Hangzhou Danjie Medical Technology Co., Ltd. commissioned Zhejiang Xianyi Pharmaceutical Co., Ltd. to trial production. When used, the contents of the capsule were poured out, and the analytical balance with an accuracy of one hundred thousandth was accurately weighed according to the test dose requirement, and carefully placed in another blank capsule. 2) Salbutamol sulfate capsules.

(3) Test method

Construction of rat COPD model by smoking combined with elastase

The use of poisoning cabinets, closed airflow. Rats were smoked at 18 per batch. Cigarette brand: Harder Gate. Passive smoking, igniting 8 cigarettes per cigarette holder, the burning time of the cigarette is about 5-6 minutes, and the sealing is kept for 30 minutes. Smoke each morning (four batches) and afternoon (four batches). The model production cycle is 12 weeks. In order to further highlight the symptoms of COPD, elastase tracheal perfusion, rat anesthesia, intratracheal elastase spray (4.8                 Units/100 g body weight, rats were injected into 15 units/0.5 mL at 300 g. Check after about 7 days, or medication.

The sampling situation is as follows:

Two normal rats (348g±5%) were selected, male and female, and two model rats, male and female. The respiratory condition of the rats was measured using a breath meter.

Rat respiratory parameters were measured and the results are shown in Table 4 below.

	Normal-F		Model-F			Normal-M		Model-M
	Mean	Standard deviation	Mean	Standard deviation	p	Mean	Standard deviation	Mean	Standard deviation	p
f	76.83	16.88	114.55	25.83	0.04	74.31	2.10	69.35	6.18	0.17
TVb	1.09	0.09	1.24	0.07	0.03	1.22	0.07	1.24	0.13	0.87
MVb	84.32	21.34	140.77	30.30	0.02	90.78	6.74	84.98	10.27	0.37
Penh	0.84	0.63	1.11	0.81	0.62	1.74	0.61	0.48	0.17	0.00
PAU	0.89	0.46	1.04	0.57	0.69	1.22	0.33	0.82	0.34	0.13
Rpef	0.23	0.30	0.10	0.02	0.41	0.08	0.01	0.06	0.02	0.01
PIFb	6.49	2.09	10.62	2.93	0.05	5.78	0.47	10.13	1.61	0.00
PEFb	5.05	0.77	9.35	0.97	0.00	7.69	0.36	6.06	0.80	0.01
Ti	0.30	0.07	0.19	0.04	0.02	0.34	0.01	0.20	0.03	0.00
Te	0.51	0.12	0.36	0.06	0.05	0.47	0.03	0.29	0.07	0.00
EF50	0.29	0.12	0.21	0.05	0.23	0.21	0.02	0.13	0.02	0.00
Tr	0.19	0.06	0.28	0.07	0.08	0.22	0.03	0.39	0.05	0.00
TB	0.94	0.74	4.64	3.86	0.45	2.52	0.69	1.73	0.39	0.42
TP	10.18	3.03	17.58	7.01	0.10	14.06	2.58	3.21	2.30	0.00

the result shows:

From the respiratory parameters, some of the basic respiratory parameters of the model mice were significantly different from those of the normal mice. The maximum expiratory flow rate PEFb and the maximum inspiratory flow PIFb of the model rats increased, suggesting that the symptoms of pulmonary hyperinflation were obvious. The model rats had faster respiratory rate and shorter respiratory time (Te) and inspiratory time (Ti). The respiratory flow rate (EP50) of 50% capacity was further reduced. It shows that the symptoms of COPD are more obvious. Further pathological examination was performed to observe the degree of pathological changes in the lung tissue.

Pathological section of rat lung tissue

After the above four rats were sacrificed, lung tissues were taken, embedded in paraffin, and stained with H&E. After dyeing, as shown in Figure 4. The results showed that a large number of inflammatory cells were found in the lung tissue of the model rats, and the dermal cells proliferated and stratified, and the goblet cells proliferated significantly. The alveolar cavity is enlarged, and most of the alveolar ruptures fuse to form large pulmonary vesicles, and the number of alveoli is significantly reduced. It has typical pathological features of COPD.

Weighing of alendronate sodium powder and HPLC calibration of powdered dose

According to the dose of rat powder for pulmonary administration, about 100-1000 ug                 Alendronate/rat, in order to accurately weigh the alendronate sodium aerosol, a reverse calibration method was used.

The small drug copy was made by using hard alloy foil, and it was used for lung administration. The fixed capacity of the drug was 1.3 mm ³ .

The alendronate sodium powder powder was continuously taken 10 times, dissolved in 1 mL of 0.2 M citrate buffer, and the content of alendronate in the solution was determined by HPLC. The results are shown in Table 5:

frequency	1	2	3	4	5	6	7	8	9	10	Mean	Standard deviation
AlN content (mg)	1.67	1.35	1.58	1.42	1.44	1.41	1.46	1.53	1.58	1.82	1.53	0.13

Conclusion: Each alendronate sodium spray contains approximately 1.53 mg ± 0.13 mg of alendronate.

In order to dose the powder of the micro-drug to the rat, the rat lung mist applicator must be calibrated to mainly calibrate the dose of the first spray.

Take 1 part of alendronate powder spray powder from the pharmaceutical and put it into the rat lung mist sprayer, and use the usual powder spraying speed for the first spray, sprayed with 1mL                 A 50 mL centrifuge tube of 0.2 M citrate buffer was mixed and the alendronate sodium content was calibrated by HPLC. Do a total of 10 times. The results are shown in Table 6:

frequency	1	2	3	4	5	6	7	8	9	10	Mean	Standard deviation
AlN content (mg)	0.53	0.49	0.64	0.34	0.53	0.48	0.34	0.42	0.24	0.42	0.32	0.12

Conclusion: The dose of the first spray of the rat lung powder mist administration kit is 0.32±0.12 mg. Rat lung administration will be carried out based on this.

Therapeutic response of a rat COPD model to alendronate (with salbutamol sulfate as a positive control)

Breathing parameter description:

sRaw: Special airway resistance.

Rpef: Pre-peak exhalation of PEF

EF50: Exhalation speed when exhaling 50% tidal volume

TV: tidal volume

Frc: Functional residual capacity

PIF: Inspiratory peak

PEF: peak expiratory

Effect of aerosol inhalation of salbutamol on respiratory function in COPD rats (S+E)

Using COPD rats (S+E: smoking + elastase modeling) as materials, respectively, aerosol inhalation 0, 0.125                 Mg/ml, 0.25 mg/ml, 0.5 mg/ml, 1 mg/ml, 2 mg/ml salbutamol-physiological saline solution, and respiratory parameters such as airway resistance were measured using a Buxco animal respiratory monitoring system. The measurement time was 5 minutes after the aerosol administration, and 150 to 250 respiratory cycles were measured. The respiratory parameters inhaled by saline were baseline (self-control), and the relative respiratory parameter changes of each rat were calculated and averaged (n=4). The result is shown in Figure 5.

Summary: After inhaling salbutamol in COPD rats, the specific airway resistance sRaw gradually decreased with the increase of drug concentration, and the final decrease was ~20%. Effective drug concentration range: 0.25 mg/mL-2 mg/mL.

Effect of aerosol inhalation of alendronate on respiratory function in COPD rats (S+E)

Using COPD rats (S+E: smoking + elastase modeling) as materials, respectively, aerosol inhalation 0,0.32mg, 0.625                 Mg, 1.25 mg, 2.5 mg, 5 mg of alendronate sodium aerosol, using Buxco animal respiratory monitoring system to detect respiratory parameters such as airway resistance. The measurement time was 5 minutes after the aerosol administration, and 150 to 250 respiratory cycles were measured. The respiratory parameters inhaled by saline were baseline (self-control), and the relative respiratory parameter changes of each rat were calculated and averaged (n=4). The results are shown in Fig. 6.

Summary: After inhalation of alendronate in COPD rats, the specific airway resistance sRaw gradually decreased with the increase of drug concentration, and the final decrease was ~40%. Effective drug concentration range: 0.625 mg/mL-5 mg/mL. Its single-agent efficacy is superior to that of salbutamol sulfate.

6. Therapeutic effect of alendronate sodium powder on salbutamol-resistant COPD rats

Therapeutic effect of alendronate sodium (ALN) on salbutamol-resistant COPD rats

Experimental design: The main airway resistance sRAW was measured before and after the induction of salbutamol resistance by using the self-control method to minimize the influence of individual model differences on the experimental results, and to investigate the effects of salbutamol tolerance on COPD rats. Does lendronate have a therapeutic response? A normal rat control group was also set up to monitor the quality of the model.

Rat grouping: Take SE (smoking + Elastase)                 Four COPD rats modeled by the method (ear label: S-1 (#53), S-2 (#52), S-3 (#51), S-4                 (#74)) As an experimental group, all were female. Another 4 normal rats of the same age (early number control-1 (#70), control-2 (#71), control-3)                 (#72), control-4 (#73), female, as a control.

Drugs: Salbutamol sulfate aerosol (Vantolin, GlaxoSmithKline, 200 spray / bottle, 50ug / spray), salbutamol original drug, alendronate sodium powder spray.

The specific experimental process is:

Day 0: Using BUXCO                 The airway sRaw value of the gradient concentration of salbutamol was determined by BFL0500 small animal respirometer for the above 4 control rats and 8 COPD rats. Salbutamol concentration gradient: 0, 0.125                 Mg/ml, 0.25 mg/ml, 0.5 mg/ml, 1 mg/ml, 2 mg/ml.

Day 1 - Day 21: Drug resistance induction

Four control rats, four COPD rats were placed in one cage, and salbutamol aerosol was given once daily, except Saturday and Sunday. Mode of administration: Cover the cage with a large plastic bag, and apply 1 spray (50 ug) to the upper part of the cage, let the rats inhale freely, fold the mouth of the bag, hold it for 5 minutes, and then give 1 spray for 5 minutes.

Day 22: The airway sRaw values of each rat in the aerosol inhalation of salbutamol were determined. Compared with the results on day 0, the rats were tested for tolerance to salbutamol after 21 days of administration of salbutamol.

Day 23:                 The airway sRaw values of albendron sodium inhalation were measured to determine whether salbutamol-tolerant COPD rats had a drug response to alendronate. Alendronate concentration gradient: 0, 0.31                 Mg/ml, 0.625 mg/ml, 1.25 mg/ml, 2.5 mg/ml, 5 mg/ml.

Experimental result

(1) Reactivity of salbutamol in normal rats and COPD rats (Day 0) before drug tolerance

The reactivity of four normal rats to salbutamol on day 0 is shown in Figure 7. The reactivity of four COPD rats to salbutamol on day 0 is shown in Figure 8. The reactivity (mean) of salbutamol on day 0 of the four normal and four COPD rats is shown in FIG.

CONCLUSIONS: Rats in the normal group also responded slightly to salbutamol, with an average sRaw value of 5-8%. COPD rats were significantly responsive to salbutamol, with an average sRaw value of 16-19%. It is indicated that the COPD rats used in this experiment are suitable for the evaluation of the efficacy of this drug.

(2) The reactivity of rats in the normal group and the COPD group to salbutamol after 21 days of salbutamol tolerance induction.

After 21 days of salbutamol tolerance induction, the reactivity of 4 normal rats to salbutamol is shown in Figure 10, after induction of salbutamol tolerance for 21 days,                 The reactivity of the rats in the 4 COPD groups to salbutamol was as shown in Figure 11, after induction of salbutamol tolerance for 21 days,                 The mean comparison of the reactivity of the rats in the 4 normal group and the 4 COPD groups to salbutamol is shown as 12. The mean values of the reactivity of salbutamol in the 4 COPD rats before and after the induction of salbutamol tolerance are shown in Fig. 13.

Summary: After 21 days of salbutamol tolerance induction, COPD rats were resistant to salbutamol. The inhalation of salbutamol did not induce an average sRaw value of 16-19%. After induction, salbutamol inhalation only resulted in a 3-5% decrease in the average sRaw value. This indicates that COPD rats are resistant to salbutamol after 21 days of induction.

(2) The reactivity of rats in the normal group and the COPD group to alendronate after 21 days of salbutamol tolerance induction.

After induction of salbutamol tolerance for 21 days,                 The reactivity of 4 normal rats to alendronate was shown in Figure 14. After induction of salbutamol tolerance for 21 days,                 The reactivity of 4 COPD rats to alendronate was shown in Figure 15, and after induction of salbutamol tolerance for 21 days,                 The mean values of reactivity of alginate sodium to alendronate in the 4 COPD rats are shown in Figure 16. Before and after induction of salbutamol tolerance, 4 normal rats and 4 COPD rats responded to alendronate. The mean comparison of the sex is shown in Figure 17.

Summary: After 21 days of salbutamol tolerance induction, COPD rats were resistant to salbutamol, but still had significant reactivity to alendronate. Salbutamol-tolerant COPD rats, treatment with alendronate can still cause an average sRaw value of 28-30%, indicating that alendronate has a different pharmacological mechanism from salbutamol and can be used in the treatment of salbutamol-resistant COPD rats. .

Example 1

Take 1000g of alendronate sodium raw material drug (produced by Shaanxi Hanjiang Pharmaceutical Group Co., Ltd.) and load it into the QS100 type pulverizer. The pulverization conditions were a crushing pressure of 0.7 MPa, and the mixture was pulverized 4 times for 3 hours to obtain a fine powder of alendronate.

The alendronate content was determined and dispensed in hard capsules at 6 mg/capsule. Laser particle size measurement results: Malvern, UK                     The Instrument Co. laser granulometer was used to determine the particle size of the capsule contents of different batches. As a result, the percentage of the sample having a particle size of less than 20 μm was greater than 90%, and the percentage of the portion having a particle size of less than 5 μm was less than 50%. The powder content of the capsule was observed by electron microscopy. Under the 10000-times SEM electron microscope, the powder was a single fine particle, no adhesion, no agglomerate, and good dispersion, and was an irregular three-dimensional structure.

In order to investigate whether the preparation process caused the destruction and degradation of alendronate sodium, the raw material of alendronate sodium before pulverization and the powder after pulverization were determined by molybdenum blue colorimetry. The method is to take 8 mg of the raw material medicine or powder, and prepare a solution having a concentration of alendronate sodium equivalent to 0.08 mg/ml, and measure the absorbance of the corresponding wavelength according to the analysis method. As a result, the absorbance of the powder after pulverization (0.071±0.007) was comparable to that of the raw material before pulverization (0.067±0.006), and it was found that the content of alendronate did not change significantly.

Example 2

Take 50 g of alendronate sodium raw material (produced by Shaanxi Hanjiang Pharmaceutical Group Co., Ltd.) and load it into an AO type pulverizer. The pulverization conditions were a crushing pressure of 0.7 MPa, 2.5 hours, and pulverization twice. A fine powder of alendronate is obtained. The alendronate content was determined and dispensed in hard capsules at 2 mg/capsule. Laser particle size measurement results: Malvern, UK                     The Instrument Co. laser granulometer was used to determine the particle size of the capsule contents of different batches. As a result, the percentage of the sample having a particle size of less than 20 μm was greater than 90%, and the percentage of the portion having a particle size of less than 5 μm was less than 50%. The powder content of the capsule was observed by electron microscopy. Under the 10000-times SEM electron microscope, the powder was a single fine particle, no adhesion, no agglomerate, and good dispersion, and was an irregular three-dimensional structure.

In order to investigate whether the preparation process caused the destruction and degradation of alendronate sodium, the raw material of alendronate sodium before pulverization and the powder after pulverization were determined by molybdenum blue colorimetry. The method is to take 8 mg of each of the above raw materials or powders to prepare a solution having a concentration of alendronate sodium equivalent to 0.8 mg/ml, and measure the absorbance of the corresponding wavelength according to an analytical method. As a result, the powder absorbance after pulverization (0.069±0.008) was comparable to the bulk drug (0.067±0.006) before pulverization, and the content of alendronate was not significantly changed.

Example 3

The alendronate sodium atomization agent prepared in Example 1 was used to measure the droplet distribution by an atomization absorption device, 12.7% was deposited in a first-order distribution bottle; and 78.2% was deposited in a secondary distribution bottle. After the application of the alendronate sodium powder of the present embodiment, the lungs of the rats were respectively subjected to HPLC analysis, and the results showed that the amount of lungs was 83.40% to 92.2%.

Example 4

The results of the rat COPD model showed that the dose of alendronate sodium aerosol inhaler was 0.6 ~ 5.0 mg, once a day, can improve the respiratory function of COPD rats ~40%, better than the effect of salbutamol sulfate (~20%) . The continuous application of the alendronate sodium powder of the present invention for 3 weeks still improves the respiratory function of COPD rats by ~30%, indicating that there is no drug resistance problem.

Example 5

For a continuous administration of salbutamol sulfate for 3 weeks, the effect was significantly decreased (respiration function improved only 3% to 5%) in rats, and the alendronate sodium powder of the present invention was dosed at a dose of 0.6.                     ~ 5.0 mg, once a day, can still further improve its respiratory function by 28% to 30%, showing that it can overcome the resistance of salbutamol sulfate.

While the present invention has been described in its preferred embodiments, the present invention is not intended to limit the invention, and the scope of the present invention may be modified and changed without departing from the spirit and scope of the invention. The scope of the claims is subject to the scope of the claims.

Claims (10)

1. Alendronate sodium powder for respiratory administration, the alendronate sodium powder is a single alendronate sodium powder, and contains no excipients.
2. The alendronate sodium powder according to claim 1, wherein the alendronate sodium in the alendronate sodium powder has a purity of ≥98%.
3. The alendronate sodium powder according to claim 1, wherein the alendronate sodium powder has a particle size distribution of d90 ≤ 20 μm, d50 ≥ 5 μm, and d10 ≤ 2.5 μm.
4. The alendronate sodium powder according to claim 1, wherein the purity of the alendronate sodium in the raw material for preparing the alendronate sodium powder is ≥98%.
5. The alendronate sodium powder according to claim 1, wherein the alendronate sodium micropowder is filled in a capsule at a charge of 0.6 mg to 5 mg, and is administered once a day.
6. The method for preparing an alendronate sodium powder according to claim 1, wherein the alendronate sodium raw material is directly pulverized and prepared without adding an auxiliary material or a solvent.
7. The method for preparing an alendronate sodium powder according to claim 6, wherein the pulverization loading amount is 50 g to 1000 g, the pulverization pressure is 0.7 MPa, the pulverization time is 2.5 h to 3.5 h, and the pulverization time is 1 or more times.
8. Use of an alendronate sodium powder according to claim 1 for the treatment of chronic obstructive pulmonary disease and/or asthma.
9. The use according to claim 8, wherein the patient with chronic obstructive pulmonary disease is resistant to a β2 receptor agonist.
10. The use according to claim 8, wherein the dose is from 0.6 mg to 5 mg per day.

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