Polymyxin B sulfate dry powder inhalation and application thereof

Abstract

The invention relates to a polymyxin B sulfate dry powder inhalation and application thereof, which can effectively solve the problem of medication for treating pneumonia and pulmonary infection caused by multi-drug-resistant gram-negative bacteria, the polymyxin B sulfate dry powder inhalation is prepared from polymyxin B sulfate, leucine and mannitol, and the preparation method comprises the following steps: dissolving polymyxin B sulfate, leucine and mannitol in ultrapure water according to a mass ratio, carrying out ultrasonic treatment to obtain a uniformly mixed medicine solution, and carrying out freeze drying to obtain the polymyxin B sulfate dry powder inhalation. The composition is simple, scientific and reasonable, the raw materials are rich, the preparation method is easy to operate, the product quality is good, the composition is administrated through aerosol inhalation, the composition is effectively used for treating pneumonia and pulmonary infection caused by multi-drug-resistant gram-negative bacteria, the composition is fast to absorb, the pulmonary drug concentration is high, the stability is good, and the composition is suitable for clinical application. The medicine is a great innovation in treatment of pneumonia and pulmonary infection caused by the multi-drug-resistant gram-negative bacteria, and has remarkable economic and social benefits.

Description

Technical Field

[0001] The invention relates to the field of medicine, and in particular to a polymyxin B sulfate powder inhaler and an application thereof. Background Art

[0002] In 2019, the World Health Organization stated in its statistical report that bacterial pneumonia and other lung infections caused by multidrug-resistant Gram-negative bacteria caused approximately 2.6 million deaths, becoming the fourth leading cause of death in the world. With the emergence of carbapenem-resistant Gram-negative bacteria (CRO) and the slow progress in the development of new antimicrobial drugs, clinical practice often falls into the dilemma of no antimicrobial drugs available. The results of CHINET China's bacterial resistance monitoring in 2019 showed that the resistance rates of Escherichia coli, Klebsiella, Pseudomonas aeruginosa, and Acinetobacter baumannii to polymyxin B sulfate in China's tertiary hospitals were 1.1%, 1.8%, 0.8%, and 0.3%, respectively. The low resistance rate of clinical isolates to polymyxin B has enabled polymyxins, which are effective against almost all CROs, to return to the clinic and be used for first-line treatment. Polymyxin B is a cyclic peptide antibacterial drug produced by Paenibacillus polymyxa. It is a narrow-spectrum antibacterial drug with strong antibacterial activity against most aerobic Gram-negative bacteria, among which Pseudomonas aeruginosa, Acinetobacter baumannii, Stenotrophomonas maltophilia and Enterobacteriales are highly sensitive to it.

[0003] At present, the main way to clinically use polymyxin B sulfate to treat lung infections is intravenous injection. However, after intravenous administration, polymyxin B must pass through barriers such as alveolar epithelial cells to be transported from the blood circulation to the alveoli and lung epithelial lining fluid to exert its effect. Studies have shown that after systemic administration of polymyxin B sulfate, the required drug concentration cannot be achieved in the lungs, and the therapeutic window of polymyxin B sulfate is narrow. Increasing the dosage is likely to cause systemic adverse reactions. Therefore, many domestic and foreign medication guidelines and expert consensus recommend nebulized inhalation of polymyxin B sulfate as one of the treatments for pneumonia caused by multidrug-resistant Gram-negative bacteria.

[0004] At present, there are no polymyxin B inhalation preparations on the market at home and abroad. The nebulized inhalation of polymyxin B recommended in medication guidelines and expert consensus refers to the use of polymyxin B injection instead of inhalation dosage forms. This non-inhalation dosage form of drug atomization can cause a variety of adverse reactions, and the antioxidants and preservatives used in the injection may also cause severe airway spasms in patients. Therefore, it is imperative to develop new drugs to effectively treat pneumonia and lung infections caused by multidrug-resistant Gram-negative bacteria. Summary of the invention

[0005] In view of the above situation, in order to overcome the shortcomings of the prior art, the purpose of the present invention is to provide a polymyxin B sulfate powder inhaler and its application, which can effectively solve the problem of medication for treating pneumonia and lung infections caused by multidrug-resistant Gram-negative bacteria.

[0006] The invention discloses a polymyxin B sulfate powder inhaler, which is prepared from polymyxin B sulfate, leucine and mannitol in a mass ratio of 1-10:1-10:1-10. The preparation method is as follows: dissolving in ultrapure water according to the mass ratio, obtaining a uniformly mixed drug solution by ultrasound, wherein the solution concentration is 8mg / mL-40mg / mL, then spraying and drying the drug solution, collecting powder, and obtaining a finished product. The drying conditions are as follows: an inlet temperature of 100°C-140°C, a spray air flow rate of 355L / h-831L / h, a feed speed of 1.2mL / min-3.6mL / min, and an exhaust air flow rate of 29m 3 / h~35m 3 / h.

[0007] The polymyxin B sulfate powder inhaler is used in the preparation of a medicine for treating pneumonia and lung infection caused by multidrug-resistant Gram-negative bacteria.

[0008] The invention has simple components, is scientific and reasonable, has abundant raw materials, is easy to operate in the preparation method, has good product quality, is administered by atomization inhalation, is effectively used for treating pneumonia and lung infection thereof caused by multidrug-resistant Gram-negative bacteria, has fast absorption, high drug concentration in the lungs, and good stability, is a major innovation in drugs for treating pneumonia and lung infection thereof caused by multidrug-resistant Gram-negative bacteria, and has significant economic and social benefits. DETAILED DESCRIPTION

[0009] The specific implementation modes of the present invention are described in detail below in combination with embodiments and specific situations.

[0010] The present invention is specifically implemented by the following examples.

[0011] Embodiment 1:

[0012] The invention discloses a polymyxin B sulfate powder inhaler, which is prepared by polymyxin B sulfate, leucine and mannitol in a mass ratio of 4:1:1. The preparation method is as follows: dissolving in ultrapure water according to the mass ratio, obtaining a uniformly mixed drug solution by ultrasound, wherein the solution concentration is 16 mg / mL, then spraying and drying the drug solution, collecting powder, and obtaining a finished product. The drying conditions are as follows: an inlet temperature of 120°C, a spray air flow rate of 667 L / h, a feed speed of 1.8 mL / min, and an exhaust air flow rate of 35 m 3 / h.

[0013] Embodiment 2:

[0014] The polymyxin B sulfate powder inhaler of the present invention is prepared from polymyxin B sulfate, leucine and mannitol in a mass ratio of 4.3:1.1:1, and the preparation method and process conditions are the same as those in Example 1.

[0015] Embodiment 3:

[0016] The polymyxin B sulfate powder inhaler of the present invention is prepared from polymyxin B sulfate, leucine and mannitol in a mass ratio of 2:X:1 to prepare an aqueous solution with a concentration of 16 mg / mL, wherein the value of X is 1-10, and the preparation method and process conditions are the same as those in Example 1. The obtained powder is characterized by using product yield, fluidity, density and the like as indicators, as shown in Table 1:

[0017] Table 1 Evaluation of PMBSDPI with different ratios of polymyxin B sulfate and leucine

[0018]

[0019]

[0020] From the results in the table, it can be seen that the yield of the formulations with different ratios is basically about 70%, which is relatively high. From the data of the angle of repose, it can be found that the angle of repose of the preparation first decreases and then increases with the increase of the leucine content. When the mass ratio of polymyxin B to leucine is 4:1, the angle of repose of the preparation is the smallest, which is 44.7±1.8°.

[0021] Embodiment 4:

[0022] The polymyxin B sulfate powder inhaler of the present invention is prepared from polymyxin B sulfate, leucine and mannitol in a mass ratio of 4:1:X to form an aqueous solution with a concentration of 16 mg / mL, wherein the value of X is 1-10, and the preparation method and process conditions are the same as those in Example 1. The obtained powder is characterized by using product yield, fluidity, density and the like as indicators, as shown in Table 2:

[0023] Table 2 Evaluation of PMBSDPI with different ratios of polymyxin B sulfate and mannitol

[0024]

[0025] From the results in the table, it can be seen that the repose angles of different formulations decrease first and then increase with the increase of mannitol content. Among them, the repose angle of M3 is 39.8±1.6°, which can meet the requirements of powder fluidity in the production process. Powder density greatly affects the aerodynamic behavior of particles. It is reported that the tap density of powder suitable for inhalation is less than 0.4g / cm 3 Among them, the repose angle of prescription M3 is the smallest and the tap density is low, which may indicate that M3 has better atomization behavior.

[0026] Example 5

[0027] The polymyxin B sulfate powder inhaler of the present invention is prepared by weighing polymyxin B sulfate, leucine and mannitol at different variable levels and different mass ratios to prepare an aqueous solution with a concentration of 16 mg / mL. The preparation method and process conditions are the same as those in Example 1, as shown in Table 3:

[0028] Table 3 Variables and levels of the star-point design experiment

[0029]

[0030] Table 4 Star point design experimental arrangement and results

[0031]

[0032] The statistical software was used to fit the quadratic regression model for the repose angle of different preparations, and the following regression equation was obtained: Y = 41.26 + 0.20 × X1 - 0.55 × X2 + 2.77 × X1 2 +2.02×X2 2 , R 2 =0.8940. The best prescription calculated by the analysis software is polymyxin B sulfate:leucine:mannitol=4.3:1.1:1 (w / w / w).

[0033] Embodiment 6:

[0034] Polymyxin B sulfate, leucine and mannitol were weighed and dissolved in water at a weight ratio of 4.3:1.1:1 (w / w / w), and aqueous solutions of different concentrations were prepared. The preparation method and process conditions were the same as those in Example 1. The concentration of the drug solution and the parameters of spray drying are shown in Table 5. A uniform experimental design was adopted, with 5 factors and 5 levels. U* 10 (10 8 )Uniform design table and quasi-horizontal method design experiment.

[0035] Table 5 Factor level design table

[0036]

[0037] According to U* 10 (10 8 ) uniform design table was used to conduct experiments, with yield, angle of repose, and fine particle fraction (FPF) in vitro as the main indicators, and the optimal process parameters were selected through the Z-score comprehensive evaluation method. The results are shown in Table 6.

[0038] Table 6 Uniform design experimental results of preparing polymyxin B sulfate powder aerosol by spray drying

[0039]

[0040] The Z-score comprehensive evaluation method is used to make the indicators dimensionless and conduct a comprehensive evaluation of each indicator. The preferred spray drying process parameters are as follows: liquid concentration 32 mg / mL, inlet temperature 120°C, spray gas flow rate 831 L / h, feed rate 3.0 mL / min, exhaust gas flow rate 33.5 m 3 / h. At this time, the fine particle fraction of the polymyxin B sulfate powder inhaler was 57.9±1.2%.

[0041] It should be noted that the above are only embodiments, which are used to illustrate the specific implementation methods of the present application, and are not used to limit the scope of protection of the present invention. All technical solutions that use equivalent and equivalent alternative technical means and are essentially the same as those of the present application belong to the scope of protection of the present application. For example, polymyxin B sulfate can also be prepared with one or more of lactose, sucrose or glycine to prepare the polymyxin B sulfate powder inhaler of the present application.

[0042] The polymyxin B sulfate powder inhaler given in the above embodiment is used in the preparation of a drug for treating pneumonia and lung infection caused by multi-drug resistant Gram-negative bacteria, and has achieved very good technical results through experiments. The relevant experimental data are as follows:

[0043] Experiment 1:

[0044] The in vivo pharmacodynamics of a single dose of polymyxin B sulfate powder inhaler was verified by constructing a rat model of acute lung infection with Pseudomonas aeruginosa. Twenty-four male SD rats were randomly divided into four groups: healthy rat group, model group, intravenous injection group (2 mg / kg) and aerosol administration group (2 mg / kg). The bacterial lung infection model was established by tracheal instillation. The specific method was to fix the rat on the intubation platform after anesthesia, insert a 20G tracheal cannula into the trachea through the mouth, and slowly instill 100 μL of bacterial suspension into the trachea with a 1mL syringe, and repeat once after 24 hours. The dry powder inhaler was administered using a pulmonary dry powder quantitative nebulizer. The rats were tested 24 hours after administration, and the number of bacteria in the lung tissue of the rats was counted by homogenization. From the colony count results of each group, it can be found that after two tracheal instillations of bacterial suspension, the number of bacteria in the lung tissue of the model group rats was 6.91±0.79lgCFU of PA per gram of lung tissue. In addition, the changes in blood routine indicators, serum PCT levels, and inflammatory factor levels in BALF and changes in lung tissue pathology jointly proved the successful construction of the acute infection model. After drug treatment, the infection of rats in the intravenous administration group was alleviated, as reflected in the reduction of lung tissue bacterial counts and the improvement of blood routine, inflammatory factors and other indicators. However, it can be seen from the experimental results that the efficacy of intravenous administration of polymyxin B sulfate in the treatment of acute lung infection is limited. On the contrary, the nebulized inhalation administration group has a good effect, which can significantly reduce the number of bacteria in lung tissue and better alleviate the abnormal indicators caused by bacterial infection. It shows that nebulized administration is better than intravenous administration. This may be because it is difficult to achieve the required lung tissue drug concentration when polymyxin B is administered intravenously, while nebulized administration can significantly increase the concentration of the drug in the lungs and thus improve the efficacy.

[0045] Experiment 2:

[0046] A rat model of chronic lung infection with Pseudomonas aeruginosa was established to verify the in vivo pharmacodynamics of multiple administrations of polymyxin B sulfate powder inhaler. The grouping and administration dosage were the same as those of the acute infection model, and the bacterial lung infection model was established by tracheal instillation. On the first day, 120 μL of agar bead suspension encapsulating bacteria was slowly instilled into the trachea using a 1 mL syringe. On the third day, 120 μL of agar beads encapsulating bacteria were instilled again. Administration began on the seventh day and continued for 7 times. Dry powder inhalation was administered using a pulmonary dry powder quantitative nebulizer. Rats were tested 24 h after the 7th administration, and the number of bacteria in the rat lung tissue was counted by homogenate. Rats in the untreated model group still had 4.96 ± 0.50 lgCFU of PA per gram of lung tissue on the 14th day. Combined with the changes in other indicators, it was proved that the chronic infection model was successfully established. After 7 consecutive days of treatment, the infection conditions of the different administration groups were significantly improved, and the improvement of the nebulization administration group was better than that of the intravenous administration group. However, judging from the situation in the nebulized drug administration group, there were still residual bacteria and abnormalities in some indicators. This may be because Pseudomonas aeruginosa forms biofilms during chronic lung infections, thus affecting the efficacy of antibiotics.

[0047] Experiments show that the present invention can be used to treat pneumonia and lung infections caused by multidrug-resistant Gram-negative bacteria, and can be used in the preparation of drugs for treating pneumonia and lung infections caused by multidrug-resistant Gram-negative bacteria, thus opening up a new way to treat pneumonia and lung infections caused by multidrug-resistant Gram-negative bacteria, with significant economic and social benefits.

[0048] The above-mentioned embodiments only express several implementation modes of the present invention, and the description thereof is relatively specific and detailed, but it should not be understood as limiting the scope of the invention patent.

[0049] In addition, it should be understood that although this specification is described in accordance with the implementation modes, not every implementation mode includes only one independent technical solution, and this description of the specification is only for the sake of clarity. It should be pointed out that for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention.

Claims

1. A polymyxin B sulfate powder inhaler is prepared from polymyxin B sulfate, leucine and mannitol in a mass ratio of 1-10:1-10:1-10. The preparation method is as follows: dissolving in ultrapure water according to the mass ratio, obtaining a uniformly mixed drug solution by ultrasound, wherein the solution concentration is 8 mg / mL to 40 mg / mL, then spray drying the drug solution, collecting the powder, and obtaining the finished product. The drying conditions are as follows: the inlet temperature is 100°C to 140°C, the spray air flow rate is 355 L / h to 831 L / h, the feed rate is 1.2 mL / min to 3.6 mL / min, and the air flow rate of the exhaust fan is 29 m 3 / h~35 m 3 / h.

2. A polymyxin B sulfate powder inhaler according to claim 1, characterized in that: The drug is prepared from polymyxin B sulfate, leucine and mannitol in a mass ratio of 4:1:

1. The preparation method is as follows: dissolving in ultrapure water according to the mass ratio, obtaining a uniformly mixed drug solution by ultrasound, and the solution concentration is 16 mg / mL. The drug solution is then spray-dried and the powder is collected to obtain the finished product. The drying conditions are as follows: the inlet temperature is 120°C, the spray gas flow rate is 667 L / h, the feed rate is 1.8 mL / min, and the air flow rate of the exhaust fan is 35 m 3 / h.

3. A polymyxin B sulfate powder inhaler according to claim 1, characterized in that: The invention is prepared from polymyxin B sulfate, leucine and mannitol in a mass ratio of 4.3:1.1:

1.

4. Use of the polymyxin B sulfate powder inhaler prepared by the method of any one of claims 1 to 3 in the preparation of a drug for treating pneumonia and lung infection thereof caused by multidrug-resistant Gram-negative bacteria.

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