TW201306847A - Compositions and uses - Google Patents

Abstract

According to the invention there is provided heparin for use in the treatment of COPD, wherein, after administration to a subject, the heparin reduces inflammation in the lungs of the subject.

Description

Composition and use

The present invention relates to diseases and abnormalities for providing pulmonary diseases (including asthma (for example, anti-steroid asthma), cystic fibrosis, idiopathic pulmonary fibrosis, non-cystic fibrotic bronchiectasis, and especially Composition of the treatment of chronic obstructive pulmonary disease (COPD).

COPD

The world's sixth leading cause of death in the COPD category. Due to increased smoking rates and demographic changes in many countries, the fourth leading cause of death is expected before 2030. In the United States, COPD is the fourth leading cause of death. In 2007, the economic burden of COPD in the United States was 42.6 billion US dollars in the cost of health care costs and losses. The largest portion of total expenditure (over 70%) is due to hospitalization costs (Sullivan S D et al, Chest 117: 5-9, 2000; Ramsey S D et al, Eur Respir J 21: 29 S-35S, 2003). This is explained by the high hospitalization costs for medical care. In the most recent medical literature, the average length of hospital stay for patients with a COPD episode ranged from 5.9 days to 12 days. Any treatment that is effective in improving the patient's respiratory tract dysfunction can enable the physician to have the patient be discharged earlier or to avoid hospitalization, thereby reducing the overall financial burden of COPD.

COPD is generally considered to be one of the most severe and disabling conditions in middle-aged and elderly patients. In the United States, 80 to 90% of COPD cases are due to smoking (medicinenet.com-COPD causes, Young et al, Eur. Respir. J. 34 (2), 2009, 380-386). COPD will also be exposed to certain dusts due to long-term exposure. Caused by the environment, for example, the manufacturing industry related to mining and powder.

COPD is an incurable disease in which chronic inflammation of the bronchial passage plays an important role. Smoking and other inhalation irritating substances continue the inflammatory reaction cycle, leading to further narrowing of the respiratory tract and allergies. Due to the development of the disease, it is difficult for the patient to remove secretions due to poor villous function. Therefore, it develops into chronic spastic cough, wheezing, and difficulty breathing. Further inflammation is exacerbated by the opportunistic bacterial colonies of the respiratory tract, which can cause obstruction of the lungs.

Acute COPD episodes occur more frequently during the winter months. An acute onset of COPD can be defined as the patient's symptoms persisting from their steady state, which is beyond normal daily changes and is acute at the outset. Acute attacks are manifested in several ways, particularly increased wheezing and/or cough, chest tightness, increased cough, reduced exercise resistance, increased fluid retention, increased fatigue, and worsening dyspnea. Although virulence factors are the cause of most seizures, exposure to allergens, contaminants, or inhaled stimulants is also relevant. The pathogenic factors known to cause acute exacerbations of COPD include: rhinovirus, influenza, parainfluenza, coronavirus, adenovirus, respiratory fusion virus, chlamydia pneumoniae, Haemophilus influenzae, Streptococcus pneumoniae, mucositis Mora Bacteria, Staphylococcus aureus, Pythium pneumoniae and Pseudomonas aeruginosa. Contaminants known to cause acute attacks include nitrogen dioxide, particulates, sulfur dioxide, and ozone.

The respiratory inflammatory response consists of macrophages on the respiratory tract and CD8 T lymphocytes and neutrophils of the respiratory tract.

Cellular infiltration dramatically changes during the onset of acute COPD. Concentration of cellular elements in the airway alveolar (BAL) compared with patients with stable COPD A large increase, resulting in the number of neutrophils, making it the main inflammatory cell of the respiratory tract.

Inflammation of the neutrophilic respiratory tract is dramatically induced by all COPD episodes. The percentage of BAL in patients with acute attacks was only 18% in BAL fluids in patients with stable COPD (Papi A et al, Am J Respir Crit Care Med 173: 1114-1121, 2006). Compared with 5% of the BAL of non-smokers and healthy smokers (Papi A et al, Am J Respir Crit Care Med 173: 1114-1121, 2006), neutrophils constitute more than all cells One and a half of the elements. The cause of this neutrophil inflammatory influx is a dramatic increase in cells in the BAL that stimulate cytokines of neutrophils and secretory cytokines in the airway epithelium.

Upregulation of cytokines caused a significant increase in human leukocyte elastase (HLE) in the respiratory epithelium. This is important because it may cause proteolytic airway damage and also stimulates excessive transmission of bronchial mucus and possibly active airway epithelial epidermal growth factor by HLE and other neutrophil globin catabolic enzymes, thereby stimulating pre-inflammatory signaling.

When a cytokine signal from the vascular space for neutrophil influx enters the lungs, the neutrophil line changes rate along the vessel wall. After the discovery of intercellular junctions between endothelial cells, neutrophils migrate from the vascular space to the respiratory tract where they adhere to the cells via intercellular adhesion molecules (ICAM) expressed on the surface of the respiratory tract. When the neutrophils move along the walls of the airways, they are activated and adhere to the airway epithelium. Neutrality in addition to indiscriminate injury to inflamed tissue White blood cells engulf invading microorganisms and particles.

Acute exacerbations of COPD have been accompanied by increased inflammation of the respiratory tract and increased systemic inflammation. The most commonly used inflammatory biomarker is C-reactive protein (CRP). CRP binds to cells, oxidized lipids, and apoptotic cells and promotes their clearance from the immune system. CRP has been shown to be associated with increased lung inflammation in stable patients with COPD.

One of the effective ways to reduce neutrophilic damage in the lungs is to delay the migration of neutrophils from the blood into the lungs before neutrophil activation and release of proteolytic enzymes and oxidants into the lung environment. One way to achieve this is by intravenous administration, which delays the migration of neutrophils into the lungs, thereby reducing the overall lung and systemic inflammation due to the onset of COPD. Today, corticosteroids are used for this purpose, but have the disadvantages of inducing muscle weakness, increasing the time of alienation, inducing osteoporosis, inducing hypertension, and inducing glucose intolerance, which in some cases leads to a diabetic state. The anti-inflammatory effect of corticosteroids on the onset of COPD is also modest, and only a modest reduction in systemic inflammation is caused by CRP measurements.

Smoking in the COPD diagnosis case is related. Smoking causes activated white blood cells to flow into the lungs, followed by degranulation and release of proteolytic enzymes. Cigarette Derivatives An oxidizing agent oxidizes alpha-1-antiprotease by oxidizing an important methionine near the active site. An imbalance between proteolytic enzyme/anti-proteolytic enzyme activity due to infusion of elastase delivered to the alveolar lung unit due to smoking, and simultaneous oxidation-passivation of alpha-1-anti-proteolytic enzymes, which is considered to be human The main cause of emphysema. Similarly, mouse models have recently developed a similar inflammatory mechanism.

The induction of emphysema requires chronic exposure to tobacco smoke (TS) for more than 6 months, which also induces inflammation of the lung tissue. Although the mouse model of chronic exposure to TS is the current gold standard for COPD, such studies are inherently time consuming, labor intensive, and expensive.

A mouse model for studying TS-induced lung inflammation enables short-term evaluation of anti-inflammatory compounds for the treatment of COPD (J. C. Fox et al., ATS, 2007). In this study, the effect of exposure of C57/B16 mice to tobacco smoke for 4 or 11 days (6 smoke/day) was investigated. This study confirmed that TS exposure for 4 or 11 days induced significant BAL cell influx, which mainly consisted of macrophages and neutrophils. BAL cell influx was not significantly different after 2, 4 or 24 hours of exposure and was generally similar after 4 or 11 days of exposure (J. C. Fox et al., ATS, 2007). In this model, BAL inflammatory cell influx can be inhibited by PDE4 inhibitors and p38 MAP kinase inhibitors. A useful endpoint is provided for the inflammatory mediators of BAL and lung tissue homogenates.

The periodic onset of COPD is the main cause of morbidity, mortality, and health care costs for patients with COPD. Patients with seizures have a poor quality of life, and the lung function measured in a healthy state and measured by forced vital capacity 1 (FEV1) decreases more rapidly.

Because of the inability to obtain curative treatment, the management of severe episodes of COPD is generally aimed at mitigating symptoms and restoring function. Pharmacological management includes the use of bronchodilators, anticholinergic agents, corticosteroids, antibiotics and methylxanthines, with oxygen therapy and non-invasive respiratory assistance.

Bronchodilators are used to treat increased shortness of breath that occurs during the onset of COPD. Inhaled beta 2 agonists are typically associated with an acute attack Use an aerosol device, a hand-held dosing agent, or a dry powder inhaler. Specific examples of β2 agonists include salbutamol, albuterol, formoterol, and terbutaline. Inhaled anticholinergic agents (such as ipratropium and thiophene) can also be used for bronchodilation, and can also be administered by aerosol devices, metered dose inhalers, or dry powder inhalers. A combination product such as ipratropium-sulphate is used to simplify drug therapy.

In the absence of major contraindications, oral corticosteroids are typically recommended in all patients with acute COPD episodes, usually in combination with other therapies. For severe attacks requiring hospitalization, prednisolone or methylprednisolone is usually used.

Corticosteroids are the mainstay of anti-inflammatory treatments, but their use in the treatment of acute exacerbations of COPD is a side effect. The β -adrenergic agonist, which acts by stimulating the β2 adrenergic receptor on the smooth muscle of the airway, acts as a bronchodilator and directly reverses the compressed airway. Non-selective anticholinergic agents such as ipratropium bromide are available as bronchodilators.

asthma

Asthma is an inflammatory disease of the lungs and respiratory tract, in which the respiratory tract is easily responsive to the stimulus and narrowed. Although airflow obstruction is characteristic of asthma, it is not considered to be COPD because lung function can be restored in asthmatic patients.

Cystic fibrosis

Cystic fibrosis (also known as CF or viscous obstruction) is a widespread disease that affects the entire body, causing progressive disability and usually premature death. About 30,000 Americans have CF, the most common genetic disease that shortens life. The term cystic fibrosis refers to characteristic scars (fibrosis) and sacs in the pancreas. Formation, first identified in the 1930s. Dyspnea is the most severe symptom and is caused by a recurrent lung infection. It is treated with antibiotics and other drugs. Although it is not cured, it eventually leads to the need for lung transplantation. Many other symptoms including sinusitis, poor growth, diarrhea, and infertility are caused by the action of CF on other parts of the body.

Today, there is no cure for the CF system. A variety of products have been provided to treat this disease, and medical needs that are not met are still high in CF patients. Treatment of lung disease in CF patients includes physical therapy to help remove mucus from the lungs, antibiotics to help control infection and avoid progressive lung damage, and DNase to remove mucus from the lungs ( DNase) I (Pulmozyme®).

Cystic fibrosis patients exhibit complex respiratory pathology involving four major factor interactions, including bacterial infection, bronchoconstriction and airway obstruction, inflammation and bronchiectasis, and finally increased mucoviscoelasticity and Mucus cilia removal ability. Each of these pathological factors has the potential to affect the extent of the other three factors. For example, increased cellular infection results in increased inflammation and bronchiectasis with concomitant increases in bronchoconstriction and airway obstruction. An increase in inflammation and bronchiectasis results in increased mucoviscoelasticity and impaired mucociliary clearance. The respiratory tract itself with a damaged mucociliary clearance mechanism causes bacterial infection and, therefore, makes this cycle permanent.

Tanning agents and antibiotic treatments for the treatment of CF are typically delivered via liquid nebulization. It is recognized that such atomized delivery systems are considered to be suboptimal for the following reasons: medication is typically limited to the clinic or home environment; inappropriate Cleaning and reusing the nebulizer system can result in unnecessary atomization of microorganisms and drugs. This susceptibility to lung infections in CF patients is a problem to be treated; and poor aerosol delivery efficiency (up to 10% of delivered drugs reach the target organ) results in prolonged dosing and delivery of the drug to the environment rather than To the patient. This environmental pollution contributes to an increased amount of antibiotics in clinical conditions.

Idiopathic pulmonary fibrosis (IPF)

Idiopathic pulmonary fibrosis (IPF) (or cryptogenic fibroalveolitis (CFA)) is a slow-progressive type of lung disease characterized by pulmonary support architecture (interstitial tissue) fibrosis.

Non-cystic fibrosis bronchiectasis

Non-cystic fibrosis bronchiectasis (bronchodilation) is a large bronchiectasis and damage caused by chronic infection and inflammation. The general cause is immunodeficiency and recurrent infection, even though some conditions appear to be idiopathic. Symptoms are chronic cough and purulent cough; some patients may have fever and difficulty breathing.

There is still a need for improved treatment for lung disease.

Summary of invention

In a first aspect of the invention, there is provided a glycosaminoglycan (eg, heparin) for use in the treatment and/or prevention of an inflammatory lung disease, optionally wherein the glycosaminoglycan is reduced following administration to a subject The subject's lungs were inflamed.

According to a further aspect of the invention, there is provided: (i) a method of treating and/or preventing an inflammatory lung disease, comprising The tester uses a glycosaminoglycan (eg, heparin), optionally, wherein, after administration, the glycosaminoglycan reduces inflammation in the lungs of the subject; (ii) glycosaminoglycan (eg, heparin) is prepared. Use of a medicament for treating and/or preventing an inflammatory lung disease, optionally, wherein the glycosaminoglycan reduces inflammation of the lungs of the subject after administration to the subject; and (iii) a kit, A glycosaminoglycan (e.g., heparin) for treating and/or preventing an inflammatory lung disease, optionally, wherein the glycosaminoglycan reduces inflammation in the lungs of the subject upon administration to the subject.

In another aspect of the invention, a glycosaminoglycan delivered by an inhalation device is provided for use in the treatment and/or prevention of an inflammatory lung disease, optionally wherein the glycosaminoglycan is polymerized after administration to the subject Sugar reduces inflammation in the lungs of the subject.

Thus, in accordance with a further aspect of the present invention, there is provided: (i) a method of treating and/or preventing an inflammatory lung disease comprising delivering a glycosaminoglycan (e.g., heparin) to a subject by an inhalation device, optionally , wherein, after administration, the glycosaminoglycan reduces inflammation in the lungs of the subject; (ii) glycosaminoglycans (eg, heparin) and inhalation devices for the preparation of a medicament for treating and/or preventing inflammatory lung diseases Use, optionally, wherein the glycosaminoglycan reduces inflammation of the lungs of the subject after administration to the subject; and (iii) a kit comprising glycosaminoglycans (eg, heparin) and an inhalation device For the treatment and/or prevention of inflammatory lung diseases, optionally, wherein the glycosaminoglycan reduces inflammation of the lungs of the subject after administration to the subject.

In another aspect of the invention, a glycosaminoglycan (eg, heparin) is provided in combination with an additional active agent for treating and/or preventing an inflammatory lung disease, optionally, wherein, after administration to a subject To reduce inflammation in the lungs of the subject. According to a further aspect of the invention there is provided: (i) a method of treating and/or preventing an inflammatory lung disease comprising administering to a subject a glycosaminoglycan (eg, heparin) and an additional active agent, optionally, Wherein the administration of the subject reduces the inflammation of the lungs of the subject; and (ii) the use of a glycosaminoglycan (eg, heparin) for the preparation of a medicament for the treatment and/or prevention of an inflammatory lung disease, Further, the active agent combination, optionally, wherein the subject is administered with a decrease in inflammation of the lungs of the subject.

In another aspect of the invention, a glycosaminoglycan (e.g., heparin) for reducing inflammation is provided.

Thus, in accordance with a further aspect of the invention, there is provided: (i) a method of reducing inflammation comprising administering a glycosaminoglycan (e.g., heparin) to a subject; (ii) a glycosaminoglycan (e.g., heparin) The use of a medicament for reducing inflammation; and (iii) a kit comprising a glycosaminoglycan (eg, heparin) for reducing inflammation.

In another aspect of the invention, a glycosaminoglycan (e.g., heparin) delivered by an inhalation device is provided for reducing inflammation.

Thus, in accordance with a further aspect of the invention, there is provided: (i) a method of reducing inflammation comprising delivering a glycosaminoglycan (e.g., heparin) to a subject by an inhalation device; (ii) a glycosaminoglycan (e.g. , heparin) and an inhalation device for the preparation of a medicament for reducing inflammation; and (iii) a kit comprising a glycosaminoglycan (for example, heparin) and an inhalation device It is used to reduce inflammation.

In another aspect of the invention, the subject to be treated is a COPD subject who is suffering from an acute COPD episode.

In another aspect of the invention, the subject to be treated is a COPD subject who is suffering from a chronic COPD episode.

In another aspect of the invention, the subject to be treated is a CF subject.

In another aspect of the invention, there is provided a pharmaceutical formulation of a glycosaminoglycan (eg, heparin) for use in the treatment and/or prevention of an inflammatory lung disease, optionally wherein, after administration to the subject, the sugar Aminoglycans reduce inflammation in the lungs of subjects.

In another aspect of the invention, there is provided a device for administering a glycosaminoglycan (eg, heparin) for the treatment and/or prevention of an inflammatory lung disease, optionally wherein the subject is administered After that, glycosaminoglycan reduces inflammation in the lungs of the subject.

Simple illustration

Figure 1 shows the total cells obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The number is reduced.

Figure 2 shows the total amount obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The number of phagocytes is reduced.

Figure 3 shows the total epithelial obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The number of cells is reduced.

Figure 4 shows the total sensation obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The decrease in the number of neutral white blood cells.

Figure 5 shows the total sensation obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The number of acid white blood cells is reduced.

Figure 6 shows the total lymph obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The number of cells is reduced.

Figure 7 shows the administration of 0.16 mg/ml, 0.5 mg/ml, 1.6 mg/ml and 5 mg compared with the control group, the saline solution and the positive control group N-acetylcysteine lysine lysinate. /ml of heparin, the change in viscoelasticity of mucus.

Figure 8 shows the administration of 0.5 mg/ml, 1.6 mg/ml and 5 mg/mg compared to 0.5 mg/ml, 1.6 mg/ml and 5 mg/ml (4-6 strips) of low molecular weight heparin. Changes in mucovisile viscoelasticity (relative rate (% of Ringer's control)) in milliliters (lanes 1-3) of undifferentiated heparin, and n-acetyl cysteinyl lysine The acid salt (7th strip) demonstrates an increase in the dose-dependent increase in mucociliary rate and compatibility with clinically relevant N-acetylcysteine lysine lysine agents.

Figure 9 shows the mean platelet count after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data confirmed that no systemic exposure was present after administration of heparin throughout the study period.

Figure 10 shows the mean active partial thrombin time (aPTT) after administration of 50 mg, 100 mg, 200 mg of undifferentiated heparin compared to lactose placebo. The data confirmed that no systemic exposure was present after administration of heparin throughout the study period.

Figure 11 shows a summary of sputum neutrophil elastase induced after administration of 50 mg, 100 mg, 200 mg of delivered undifferentiated heparin compared to lactose placebo. The data confirmed that the amount of neutrophil elastase decreased after the 100 mg dose.

Figure 12 shows an overview of induced total cell counts after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data demonstrates a decrease in total cell count after administration of undifferentiated heparin compared to lactose placebo.

Figure 13 shows an overview of induced 痰IL-8 after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data demonstrates a reduction in IL-8 levels after administration of a 100 mg dose of heparin compared to lactose placebo.

Figure 14 shows a summary of plasma neutrophil elastase following administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data confirmed no plasma inflammatory markers.

Figure 15 shows an overview of plasma neutrophil counts after administration of 50 mg, 100 mg, 200 mg of delivered undifferentiated heparin compared to lactose placebo. The data confirmed no plasma inflammatory markers.

Figure 16 shows plasma NE/AAT mismatch after delivery of 50 mg, 100 mg, 200 mg of undifferentiated heparin compared to lactose placebo. Overview of things. The data confirmed no plasma inflammatory markers.

Figure 17 shows a summary of plasma CRP after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data confirmed no plasma inflammatory markers.

Figure 18 shows a summary of plasma IL-8 after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data confirmed no plasma inflammatory markers.

Figure 19 shows a summary of plasma IL-6 after delivery of 50 mg, 100 mg, 200 mg of undifferentiated heparin compared to lactose placebo. The data confirmed no plasma inflammatory markers.

Figure 20 shows a summary of plasma inflammatory mediators after administration of 50 mg, 100 mg, 200 mg delivered doses of undifferentiated heparin compared to lactose placebo. The data confirms that there is no plasma inflammatory marker amount for all neutrophils, neutrophil elastase neutrophil elastase/ α -1 antitrypsin complex, IL-8, IL-6 and CRP. In Fig. 20, the first strip in the histogram indicates a low dose, the second strip indicates a medium dose, and the third strip indicates a high dose.

Figure 21 shows mucus observations after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo, from baseline to week 4 of the study. The data confirmed that the patient's mucus became thinner after heparin treatment. The volume of sputum increased for the placebo system. The mucus color becomes clearer, indicating a reduction in bacterial infection.痰 becomes less viscous and therefore easier to remove from the lungs.

Figure 22 shows the change from the baseline to the 4th week of the study, with lactose The amount of S. aureus and Pseudomonas aeruginosa in the sputum after administration of 50 mg, 100 mg, and 200 mg of undifferentiated heparin compared to placebo. The data confirmed no microbial action.

In each of the histograms of Figures 21 and 22, if present, the first strip represents a low dose, the second strip represents a medium dose, the third strip represents a high dose, and the fourth strip represents a placebo.

Detailed description of the invention

We have found that anticoagulant heparin reduces inflammation (eg, lung neutrophil count) to an amount achieved using the p38 MAP kinase inhibitor (ADS115398). In addition, we have found that heparin also reduces inflammation in clinical settings (eg, for CF patients).

The present invention provides a method for treating the symptoms of inflammatory lung diseases including asthma, cystic fibrosis, idiopathic pulmonary fibrosis, non-vesicular fibrotic bronchiectasis, and particularly chronic obstructive pulmonary disease (COPD). . Such methods include administering heparin to a patient having an inflammatory lung disease such as asthma, cystic fibrosis, idiopathic pulmonary fibrosis, non-cystic fibrotic bronchiectasis, COPD, or any combination thereof. Administration of heparin is particularly beneficial for reducing inflammation in the lungs, and is hereby demonstrated by a reduction in the amount of lung neutrophils in the p38 MAP kinase inhibitor ADS115398.

Drugs that reduce inflammation of the lungs (eg, neutrophils), bacterial infections, and mucus viscosity are of great benefit in the treatment of CF or, in particular, COPD episodes. Heparin binds to these desired effects and avoids the need to administer multiple drugs in patients with CF or especially COPD.

The term "inflamed lung disease" is a disease associated with inflammation of the respiratory system (eg, the lungs).

The present invention demonstrates that administration of glycosaminoglycans (e.g., heparin) is particularly advantageous for reducing inflammation in the lungs (e.g., by reducing the amount of neutrophil in the lungs to be achieved by the p38 MAP kinase inhibitor ADS115398).

Embodiments of the invention that may be used alone or in any combination include those in which: (a) the glycosaminoglycan is selected from the group consisting of chondroitin sulfate, dermatan sulfate, keratin sulfate, acetaminophen sulfate, Hyaluronic acid and especially heparin (eg, glycosaminoglycans are selected from the group consisting of danaparoid sodium, dermatan sulfate, and particularly heparin); (b) administration of glycosaminoglycans (eg, heparin) results in at least 10% (eg, Inflammation reduction of at least 20%, 30%, 40%, 50% or especially 60%) (eg, measured by an inflammatory marker, as described herein (eg, total sputum cell count, neutrophil elastase amount) (c) glycosaminoglycans (eg, heparin) are administered intranasally or by pulmonary inhalation (eg, heparin is administered by pulmonary inhalation); (d) glycosaminoglycans (eg, Heparin is in the form of a dry powder formulation; (e) glycosaminoglycan-derived heparin; (f) heparin-based undifferentiated heparin (eg, undifferentiated heparin sodium, such as undifferentiated heparin sodium oxidized or unoxidized) For example, unoxidized undifferentiated heparin sodium); (g) heparin low molecular weight heparin (eg, enoxaparin sodium); (h) sugar Polysaccharides (e.g., heparin) of the total daily dose (FPD to measure) based Between 0.5 mg and 6000 mg (for example, the total daily dose of heparin is between 1 mg and 2000 mg, such as between 2 mg and 1500 mg, between 6 mg and 600 mg, at 12 mg Between 300 mg, such as between 18 mg and 240 mg, between 24 mg and 180 mg, such as between 36 mg and 120 mg (eg, between 50 and 75 mg); i) The total daily dose of heparin (measured in FPD) is between 0.5 mg and 6000 mg (for example, the total daily dose of heparin is between 1 mg and 2000 mg, such as between 2 mg and 1500 mg Between 6 mg and 600 mg, between 12 mg and 300 mg, such as between 18 mg and 240 mg, between 24 mg and 180 mg, such as between 36 mg and 120 mg. (eg, between 50 and 75 mg)); (j) Inflammation is measured by total cell count obtained from induced sputum or in particular bronchoalveolar perfusion; (k) inflammatory is measured by an inflammatory marker (eg, Inducing neutrophil elastase, inducing sputum IL-8 or especially eosinophilic white blood cell volume, lung The amount of cells, the amount of macrophages in the lungs, or more particularly the amount of neutrophils in the lungs, or any combination of these (for example, the amount of macrophages in the lungs or especially the amount of neutrophils in the lungs); (1) The achieved reduction in inflammation is equal to that achieved by the p38 MAP kinase inhibitor ADS115398 (eg, wherein ADS115398 is provided to the subject at a dose of 0.1 mg/kg); (m) COPD is an acute COPD episode; (n) COPD is a chronic COPD episode; (o) Heparin is delivered at a nominal dose between 0.8 mg and 2100 mg (for example, between 1.5 mg and 1100 mg, between 3 mg and 530 mg, between 8 mg and 300 mg, between 25 mg and 200 mg, between 35 mg and 150 mg, at 70 (mg) between heparin and between 25 mg and 200 mg, between 50 mg and 125 mg, such as between 75 and 100 mg or especially 0.5 mg and 2000 mg Between (for example, between 1 mg and 1000 mg, between 2 mg and 500 mg, between 6 mg and 250 mg, between 18 mg and 240 mg, between 30 mg and 130 mg, Fine particle dose delivery between 60 mg and 120 mg), for example, when measured by New Generation Impactor (Ph Eur Apparatus, 60 liters/min); (q) inflammatory lung disease characterized by neutrophils The disease; (r) an inflammatory lung disease may be selected from the group consisting of asthma, cystic fibrosis, idiopathic pulmonary fibrosis, non-cystic fibrosis bronchiectasis, and especially chronic obstructive pulmonary disease (eg, Persistent asthma, severe persistent asthma, cystic fibrosis, idiopathic pulmonary fibers Non-cystic fibrotic bronchiectasis and especially chronic obstructive pulmonary disease, such as persistent asthma, cystic fibrosis and especially chronic obstructive pulmonary disease (eg, cystic fibrosis and especially chronic obstruction) Disease or condition of a pulmonary disease).

(s) the use system further comprises another active agent (for example, the further active agent is selected from the group consisting of phlegm agents (for example, N-ethylcysteine, cough, amlorin, dextran, heparin, Desulfated heparin, low molecular weight heparin, and recombinant human DNase); bronchodilators (eg, β2-agonist Bambuter, Bitotrol, Bromoxate, Cammotrol, Clenbuterol, non- Notro, Formoterol, Indacaterol, L-Salbutamol, Isoprogestin, Oxylinine, Picotero, Pibuterol, Necatero, Riptro, Limitero , salbutamol, salmeterol, tert-butyl, etc.); scorpion toxin receptor blockers (eg, ipratropium, ipratropium bromide, oxitropium, tiotropium and glucose pyridine); antibiotics and antibacterial agents (for example, including β -lactam, fluorinated quinolinone, ketolide, macrolide, thiamine and tetracycline, arubicin, amoxicillin, amphotericin, azithromycin, aztreonamidine , clarithromycin, clindamycin, sulphococcin, nascentin, dirithromycin, doripenem, erythromycin, Fusarium , Jiandamycin, metronidazole, mupirocin, natamycin, neomycin, nystatin, oleandomycin, pentamidine, streptavidin, probenecid, roxithromycin, Sulfadiazine and triclosan); anti-infective agents (eg, antiviral agents (including nucleoside and non-nucleoside reverse transcriptase inhibitors, and proteolytic enzyme inhibitors), including Apollo, Adelphi, Oman Teddy, silodofur, effluent, famciclovir, foscarnet, ganciclovir, iodine, indinavir, isoproterenol, lamivudine, mefenac , nevirapine, oseltamivir, palivizumab, penciclovir, pluronic, ryba, rimantadine, ritonavir, ruplin, saquinavi , stavudine, valacyclovir, zalcitabine, zanamivir, zidovudine and interferon); aminoglycosides (eg, toxomycin; antifungal agents, eg, amylase) , caspofungin, tribendazole, econazole nitrate, fluconazole, itraconazole, clonazole, miconazole, nitsidine, terbinafine and voriconazole; anti-tuberculosis agents , for example, a volume Aflatoxin, ciprofloxacin, ethambutol, meropenem, piperacillin, rifampicin and pancomycin; β -endoamine, including cefazolin, cefmetazole, cefoperazone, cefoxit Ding, cefotaxime, cefotaxime, ceftriaxone and cephalosporin; cephalosporin, including cephalosporin C and cephalosporin; cephalosporin, such as, cephalosporin A, cephamycin B, cephamycin C, cefpirin and cefradine); anti-leprosy drugs (eg, clofazimine; penicillin, including ammonixine, ampicillin, penicillin, azidocillin, benzyl mexicillin, carbenecillin, Carfilcillin, carbocillin, clindacillin, cloxacillin, cyclohexillin, dakocillin, biphenylpenicillin, heptyl penicillin, hetacillin, metancillin, methicillin, nafcillin, 2- Pentenyl penicillin, penicillin N, penicillin O, penicillin S and penicillin V; quinolinone, including ciprofloxacin, clinfloxacin, difloxacin, gepafloxacin, norfloxacin, ofloxacin and Temasalin); tetracycline, including doxycycline and oxytetracycline; All sorts of anti-infective agents, e.g., linezolid, trimethoprim and sulfamethoxazole. Non-steroidal anti-inflammatory agents (eg, aceclofenac, acetaminophen, aminoxine, amfenac, ampicillin, amoxicillin, aspirin, phenoxaprofen, bromfenac, butyl hydroxy acid, Carprofen, celecoxib, choline, cinchon, indomethacin, clopidogrel, clonin, to be kefinal, to be kefinal, diflunisal, o-ethoxybenzoic acid, relying on Acid, etoricoxib, fenoprofen, flurbiprofen, ibuprofen, indomethacin, indoprofen, ketoprofen, ketorolac, loxoprofen, equine, Mechloric acid, mefenamic acid, meloxicam, nametidinone, naproxen, nimesulide, parecoxib, phenylbutazone, piroxicam, piroxine, ro Fenoxib, salicylate, sulindac, tirolidine, tolfenamide, tolmetine and valdecoxib; other anti-inflammatory agents (eg, B-cell inhibitors, p38 MAP kinase inhibitors, In particular, ADS115398 and TNF inhibitors; PDE4 inhibitors (eg, tetrazolast, eptazolate, rolipram, omiester, roflumilast, ONO 6126, tolafen and Zadawi) Lin) Diketones (for example, quinoxaquinone and quinquinone analogs; xanthine derivatives such as, for example, decanoicin and arophylline; tetrahydropyrimidinone, such as atrazine; and guanidinium carbamate , for example, non-Ministry; steroids (eg, aclomethasone, beclomethasone dipropionate, beclomethasone dipropionate dipropionate, betamethasone, budesonide, butoterol, ciclesonide , clobetasol, difluxate, diflupredil, deoxymethasone, dexamethasone, fludrocortisone, flunisolide, fluocinolone, flumethalin, fluticasone, fluticasone propionate, hydrogenation Cortisone, methylprednisolone, mometasone, nantrolide, neomycin sulfate, prednisolone, rimexolone, rofluronide, triamcinolone and triamcinolone acetonide Matrix metalloproteinase inhibitors (eg, metal hydrolyzed protein, serranosin and astaxanthin); epithelial sodium channel (ENaC) inhibitors (eg, P-680 and denufosol; CFTR enhancer) , for example, VX-809); methylxanthine (eg, caffeine, theobromine, and theophylline); drugs for cystic fibrosis management (eg, E.g., Pseudomonas aeruginosa vaccines (e.g., Aerugen ^TM), α 1- antitrypsin, amikacin, have blessed repair, denufosol, duramycin, glutathione, mannitol, and tobramycin Mycin).

Further embodiments of the invention, which may be used herein alone or in any combination, include wherein: (i) the total daily dose of heparin (measured in FPD) is between 0.1 and 6000 mg, such as Between 0.2 and 3000 mg (for example, between 0.3 and 250 mg, such as between 0.32 and 200 mg, such as between 1 and 175 mg, between 5 and 150 mg, such as at 10 with Between 100 mg, such as between 20 and 80 mg (eg, between 30 and 70 mg, such as between 35 and 50 mg (eg, between 40 and 45 mg)); (ii The total daily dose of heparin (measured in nominal doses) is between 1 and 8000 mg, such as between 2 and 5000 mg (eg, between 15 and 300 mg (eg, between 20 and 300 mg) Between), for example, between 25 and 250 mg (for example, between 31 and 250 mg, or between 30 and 200 mg), such as between 48 and 248 mg, such as between 62 and 150 mg. Between (for example, between 70 and 130 mg, such as between 80 and 125 mg, for example between 90 and 124 mg)); (iii) total daily dose of heparin (measured as delivered dose) between 0.5 and 6000 mg, such as between 0.8 and 5000, such as between 10 and 1000 mg (eg, between 15 and 500 mg, such as at 25) Between 400 mg, between 50 and 300 mg, between 75 and 200 mg (for example, between 100 and 150 mg); (iv) Inflammation is selected from neutrophil elastase and total The inflammatory label of the cell count is measured; (vi) heparin is delivered at a nominal dose between 0.8 mg and 500 mg (eg, between 1.5 mg and 400 mg, between 3 mg and 200 mg, Between 8 mg and 150 mg, between 10 mg and 100 mg, between 15 mg and 75 mg, between 20 mg and 31 mg), for example. When measured by New Generation Impactor (Ph Eur Apparatus, at 60 liters per minute); (vii) heparin is between 0.3 and 400 mg, between 0.5 and 200 mg, such as between 1 and 150 mg. Between, or especially between 1.5 mg and 100 mg (for example, between 5 mg and 80 mg, between 10 mg and 60 mg, between 15 mg and 40 mg, at 20 mg and 30 mg) Fine particle dose delivery, for example, when measured by New Generation Impactor (Ph Eur Apparatus, at 60 liters per minute); (viii) total daily dose of heparin (measured by FPD) at 0.1 and 6000 mg Between, for example, between 0.2 and 3000 mg (eg, between 0.3 and 250 mg, such as between 0.32 and 200 mg, such as between 1 and 175 mg, between 5 and 150 mg) , for example, between 10 and 100 mg, such as between 20 and 80 mg (eg, between 30 and 60 mg, such as between 35 and 50 mg (eg, between 40 and 45 grams) )), wherein the patient with CF did not suffer from increased hemoptysis after administration of the dose; (ix) the total daily dose of heparin (in Dose measurement) is between 1 and 8000 mg, such as between 2 and 5000 mg (eg, between 15 and 300 mg, such as between 31 and 250 mg, such as between 48 and 248 mg) Between, for example, between 62 and 150 mg (eg, between 70 and 130 mg, such as between 80 and 125 mg, for example between 90 and 124 mg), wherein, with CF The patient did not suffer from increased hemoptysis after administration of the dose; (x) The total daily dose of heparin (measured as the delivered dose) is between 0.5 and 6000 mg, such as between 0.8 mg and 5000, such as between 10 and 1000 mg (eg, between 15 and 500) Between milligrams, for example, between 25 and 400 milligrams, between 50 and 300 milligrams, between 75 and 200 milligrams (eg, between 100 and 150 milligrams), wherein a patient with CF is cast The dose is not subjected to increased hemoptysis after the dose; (xi) one or more (eg, two to eight (such as one to four) nominal doses may be delivered sequentially. It may be used alone or in any combination as described herein. Further embodiments of the invention include wherein: (i) the total daily dose of heparin (measured in FPD) is between 30 and 50 mg (eg, 40 mg) for patients with COPD; (ii) heparin The total daily dose (measured as a nominal dose) is between 90 and 110 mg (eg, 100 mg) for patients with COPD; (iii) total daily dose of heparin (measured as delivered dose) for patients with COPD Is between 115 and 135 mg (eg, 124 mg). It can be used alone or in any combination as described herein. Further examples include those in which: (i) the total daily dose of heparin (measured in FPD) is between 30 and 50 mg (eg, 40 mg) for patients with CF; (ii) total heparin The daily dose (measured as a nominal dose) is between 90 and 110 mg (eg, 100 mg) for patients with CF; (iii) the total daily dose of heparin (measured as delivered dose) for patients with CF Between 115 and 135 mg (eg, 124 mg); (iv) The total daily dose of heparin (measured in FPD) is between 30 and 50 mg (eg, 40 mg) for patients with CF, wherein the patient is not suffering from cough and/or after administering the dose. Not suffering from increased hemoptysis; (v) total daily dose of heparin (measured in nominal doses) for patients with CF between 90 and 110 mg (eg, 100 mg), wherein the patient is dosed The latter is not suffering from cough and/or does not suffer from increased hemoptysis; (vi) the total daily dose of heparin (measured as delivered dose) is between 115 and 135 mg (eg, 124 mg) for patients with CF, Among them, the patient did not suffer from coughing and/or did not suffer from increased hemoptysis after administration of the dose.

Further embodiments of the invention, which may be used herein alone or in any combination, include wherein: (a) a glycosaminoglycan formulated in a dry powder is delivered as a dry powder inhaler; (b) a dry powder inhalation The device is selected from the group consisting of passive (Rotahaler and Diskhaler, GyroHaler, Turbohaler, Novolizer or especially Monohaler, and devices manufactured by Vectura (as covered by International Patent Application No. WO 2010/086285), or active devices (Aspirair ( Trademarks) and active inhaler devices manufactured by Nektar Therapeutics (as covered by U.S. Patent No. 6,257,233) (for example, dry powder inhalers are selected from GyroHaler or, in particular, Monohaler and passive devices manufactured by Vectura (by international patents) Application No. WO 2010/086285)); (c) dry powder inhaler is a type comprising a casing and a bubble support element The inhaler of the device has a nozzle through which a user can inhale a dose of the drug, the bubble supporting member having a groove for containing a medicament containing bubbles, the outer casing and the bubble supporting member Pivoting relative to each other between the first position and the second puncture position, the first position is for inserting a bubble into the groove, and in the second puncture position, a bubble piercing element is carried by the outer casing to pierce the embedded bubble So that when the user inhales on the nozzle, the medicament is input into the airflow, and flows out of the bubble through the nozzle and flows into the airway of the user.

(For example, the device, wherein the outer casing comprises a substantially cylindrical chamber having an inlet at one end and an outlet at an opposite end thereof for self-piercing the air carrying the drug The air bag flows into the chamber for discharging the drug-loaded air from the nozzle and into the patient's respiratory tract (eg, such that the substantially cylindrical chamber has at least one bypass air inlet for cleaning Air flows into the cyclone chamber and interacts with the air carrying the negative drug flowing between the inlet and the outlet)).

Based on 60 kg subjects, human doses of 25 mg, 50 mg, and 100 mg correspond to approximately 417 μg/kg, 833 μg/kg, and 1667 μg/kg (recommended by the FDA).

Therefore, the dose of heparin in 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg is equivalent to 6 mg, 18 mg, 60 mg, and 60 μg/kg, and subject. 120 mg human dose (recommended by the FDA).

definition

The term chronic obstructive pulmonary disease ("COPD") is used to describe the general It is a respiratory disease that is partially or completely blocked by the airway that is not completely reversible. This condition can also be referred to as chronic obsessive air disease (CORD), chronic obstructive airway disease (COAD), chronic obstructive pulmonary disease (COLD), or chronic airway restriction (CAL). The word COPD is intended to include all such allegations that include the above-mentioned symptoms without limitation.

The American Thoracic Society (ATS) defines COPD as a disease involving developmental chronic airflow obstruction. The COPD system is defined as a disease state in which the airflow is not fully reversible. This airflow is limited by its progression and is generally degenerative and is associated with an abnormal inflammatory response in the lungs. The basic conditions leading to COPD include chronic bronchitis, emphysema, and bronchiectasis. Chronic bronchitis is defined as excessive coughing and spasm for at least three consecutive months for at least two consecutive years. Emphysema is characterized by chronic dyspnea (short breathing) caused by damage to lung tissue. Bronchiectasis is atypically tightened in the respiratory tract. The weakened airway becomes scarred, causing more mucus and bacteria to accumulate, thus causing a recurrence cycle that blocks the respiratory tract and infection.

Any treatment or/and prevention of COPD herein includes any of the basic conditions (ie, chronic bronchitis, emphysema, and bronchiectasis) associated with the treatment and/or prevention as described above, regardless of each other. Or combination).

For the avoidance of doubt, in the context of the present invention, the term "treatment" includes therapeutic or palliative treatment of a patient in need of such treatment, and prophylactic treatment and/or diagnosis of a patient susceptible to the relevant disease state.

The term "patient" includes patients with mammals (eg, humans).

The term "moderate persistent asthma" and "severe persistent asthma" (along with intermittent and mild persistent asthma) are used to classify the severity of asthma. this The classification of the terms can be found in Yawn, BP (September 2008), "Factors accounting for asthma variability: achieving immediate symptom control for individual patients". Primary Care Respiratory Journal 17(3): 138-147.

The term "effective amount" refers to the amount of a compound that is administered to a treating patient (eg, sufficient to treat or prevent the disease). The effect can be objective (ie, can be measured by some test or marker) or subjective (ie, the subject refers to or feels the effect).

As used herein, the term "nominal dose" (ND) is the amount of drug (also known as a metered dose) measured in a container. This is different from the amount of drug delivered to the patient as a delivered dose.

Fine particle fraction (FPF) is generally defined as the "fine particle dose" (FPD, this dose is <5 μm) divided by the dose (ED) of the dose leaving the device. FPF is expressed as a percentage. Here, the FPF of ED is called FPF (ED) and is calculated by FPF (ED) = (FPD / ED) x 100%.

FPD can be measured by the Multistage Liquid Impinger, United States Pharmacopoeia 26, Chapter 601, Apparatus 4 (2003), an Andersen Cascade Impactor or a New Generation Impactor.

As used herein, the term "fine particle fraction" (FPF) can also be defined as the FPD divided by the quantitative dose (MD) (which is the dose within the bubble or capsule) and is expressed as a percentage. Here, the FPF of MD is called FPF (MD) and is calculated by FPF (MD) = (FPD / MD) x 100%.

The term "ultrafine particle dose" (UFPD) is used here to mean that there is not much The total mass of active material delivered by the device at a diameter of 3 μm. The term "ultrafine particle fraction" is used herein to mean the percentage of the total amount of active material delivered by the device having a diameter of no more than 3 μm. The ultrafine particle dose percentage (% UFPD) is used herein to mean a percentage of the total dose delivered (ie, UPUD = 100 x UFPD / total dose) having a diameter of no more than 3 μm.

The terms "delivery dose" and "emission dose" or "ED" are used interchangeably herein. These are measured as shown in the topic of inhalation products of current European Pharmacopeia (EP).

"Inhalation start" refers to the procedure by which a dose of powder is removed from its resting position within the inhaler. This step occurs after the powder has been loaded into the inhaler for use.

The particle size is geometric unless otherwise indicated.

Embodiments of the invention, which may be used alone or in any combination, include the following: (a) a glycosaminoglycan (e.g., selected from the group consisting of chondroitin sulfate, dermatan sulfate, keratin sulfate, acetaminophen sulfate) , hyaluronic acid and, in particular, a compound of heparin) may be in the same composition as the other inhaled therapeutic agent, or in particular, heparin is separated from the composition comprising the composition of the inhaled therapeutic agent; (b) glycosaminoglycan (for example, a compound selected from the group consisting of chondroitin sulfate, dermatan sulfate, keratin sulfate, acetamidine heparin, hyaluronic acid, and special heparin) is delivered by pulmonary inhalation; (c) glycosaminoglycan (for example, selected from chondroitin sulfate, dermatan sulfate Salt, keratin sulfate, acetoin sulfate, hyaluronic acid and special heparin compounds are in the form of a composition such as a dry powder composition; (d) comprises a glycosaminoglycan (for example, selected from chondroitin sulfate) The composition of dermatan sulfate, keratin sulfate, acetamidine heparin sulfate, hyaluronic acid, and particularly heparin comprises at least 5% by weight (eg, at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%) heparin, for example, at least about 75%, 85%, 95%, 96%, 97%, 98% or 99% by weight of glycosaminoglycan A sugar (for example, a compound selected from the group consisting of chondroitin sulfate, dermatan sulfate, keratin sulfate, acetamidine heparin sulfate, hyaluronic acid, and particularly heparin).

(e) a composition comprising a glycosaminoglycan (for example, selected from the group consisting of chondroitin sulfate, dermatan sulfate, keratin sulfate, acetaminophen sulfate, hyaluronic acid, and particularly heparin) further comprising an additive material (for example, Magnesium stearate or leucine, preferably L-leucine; (f) glycosaminoglycan (for example, selected from the group consisting of chondroitin sulfate, dermatan sulfate, keratin sulfate, acetaminophen sulfate) The salt, hyaluronic acid and especially heparin composition further comprises one or more excipient materials (eg, inorganic salts, organic salts, other organic compound sugars, or more particularly alcohols, polyols, and crystalline sugars (such as mannitol). Carrier particles made of trehalose, melezitose, dextrose, or especially lactose; (g) carrier particles may be present between 5 and 1000 μm (eg, 4 to 500 μm, such as, Average particle size of 20 to 200 μm, 30 to 150 μm, 40 to 70 μm, or 60 μm; (h) glycosaminoglycan heparin; (i) the maximum daily dose of heparin is less than 6000 mg (eg, less than 2000 mg, such as less than 1500 mg, 600 mg, 300 mg, 240 mg, 180 mg, especially 120 mg); (j) A dose of heparin is provided in a fine particle dose of heparin between 0.5 mg and 2000 mg (eg, between 1 mg and 1000 mg, between 2 mg and 500 mg, between 6 mg and 250 mg, Between 18 mg and 240 mg, between 30 mg and 130 mg, between 60 mg and 120 mg), for example, when measured by New Generation Impactor (Ph Eur Apparatus, at 60 liters/min); (k) The anti-inflammatory effect of heparin depends on the dose (for example, heparin is determined by the dose of BAL in neutrophils and macrophages after heparin treatment); (1) Heparin is administered 120 The therapeutic effect is provided within minutes (eg, between 30 and 60 minutes); (m) the heparin system is administered sequentially, simultaneously or concomitantly with additional active agents.

Embodiments of the invention, which may be used alone or in any combination, include the following: (a) The maximum daily delivery dose of heparin is less than or equal to 2000 mg (eg, less than or equal to 500 mg, less than or equal to 300) Mg, 200 mg, 150 mg, 100 mg, 80 mg, 40 mg, 20 mg, 10 mg, 5 mg, or 1 mg); (b) One dose of heparin is between 0.3 and 400 mg at 0.5 200 mg, for example between 1 and 150 mg, or especially between 1.5 mg and 100 mg (for example, between 5 mg and 80 mg, between 10 mg and 60 mg, Provide a fine particle dose of heparin between 15 mg and 40 mg between 20 mg and 30 mg), for example, when measured by New Generation Impactor (Ph Eur Apparatus, at 60 liters per minute); (c) An additive of between 1 and 20% by weight (eg, between 2 and 15% by weight, between 5 and 10% by weight, between 7 and 9% by weight) (eg, between 3 and 15%, at 5 and 10%) A heparin dry powder formulation of between 7 and 9% leucine, such as L-leucine.

In one embodiment, the nominal dose of heparin administered to the patient is up to 2100 mg, 1100 mg, 530 mg, 300 mg or especially up to 200 mg, in particular, the nominal dose is at least 100 g or especially 0.8 mg. 1.5 mg, 3 mg, 8 mg, 25 mg, 35 mg, 70 mg, and 140 mg).

In another embodiment, the nominal dose of heparin administered to the patient is up to 2100 mg, 1100 mg, 530 mg, 300 mg, or especially up to 200 mg, in particular, the nominal dose is at least 0.8 mg, 1.5 mg. , 3 mg, 8 mg, 25 mg, 35 mg, 70 mg, 100 mg, 140 mg, and 200 mg).

Mechanism

While not wishing to be bound by theory, it is believed that glycosaminoglycans (eg, heparin, such as undifferentiated heparin, such as undifferentiated heparin sodium) possess anti-inflammatory activity. This activity is thought to be produced by heparin, which is a large negatively charged molecule that binds to several physiological substances involved in the inflammatory procedure, such as L- and P-selectin (which reduces the attachment of blood vessel walls to white blood cells and inflamed sites). ), cytokines, factors of birth, cytotoxic peptides, and tissue disrupting enzymes. Heparin The polyanionic force of the glycosaminoglycan (GAG) chain is caused by the electrostatic force between the positively charged amino acids of the inflamed protein.

Heparin is also disclosed to reduce the total amount of activated neutrophils, which can help reduce inflammation. Neutrophilic white blood cells are a powerful source of oxygen-derived free radicals involved in the enzyme mismatched nicotinamide adenine dinucleic acid phosphate (NADPH) oxidase.

In addition, heparin has the properties of phlegm, anti-bronchial contraction and anti-infection, which is believed to help reduce overall inflammation by reducing symptoms or inflammatory causes. For example, effective removal of mucus prevents the accumulation of bacteria in the lungs, which can result in reduced inflammation by reducing the immune response.

Heparin composition for pulmonary inhalation

In the past, many commercially available dry powder inhalers exhibited very poor drug efficiencies, and sometimes as little as 10% of the active agent present in the agent was actually delivered to the user properly, making it therapeutically effective. This inefficiency is completely unacceptable when a high dose of active agent is required to achieve the desired therapeutic effect.

The lack of efficiency of administration is due to the fact that the active agent in the dry powdered medicament is readily lost from the expulsion of the delivery device to each stage of deposition in the lung. For example, a large amount of material will remain in the bubble/capsule or device. The material will be lost to the throat of the subject due to excessive plume rate. However, it is the case that a high ratio of delivered agents is present in a particle form that exceeds the aerodynamic diameter required.

It is known that the particle impaction of the upper respiratory tract of a subject is predicted by the so-called impact parameters. The impact parameter is defined as the particle velocity multiplied by its aerodynamic diameter. Squared. Thus, the probability associated with delivery of the particles through the upper respiratory tract region to the target site of action is related to the square of its aerodynamic diameter. Thus, delivery to the lower respiratory tract or deep lung is dependent on the square of its aerodynamic diameter, and smaller aerosol particles are more likely to reach the user's target site of administration and, therefore, have the desired therapeutic effect.

Particles having an aerodynamic diameter of less than 10 μm are easily deposited in the lungs. Aerodynamic particles having a range of 2 μm to 5 μm are generally deposited in the respiratory bronchi, while smaller particles having an aerodynamic diameter in the range of 0.05 to 3 μm may deposit in the alveoli. Thus, for example, the high drug efficiency of the particles targeted to the alveoli is expected to be by agents of particles below 3 μm, and smaller particles are most likely to reach this target position.

In one embodiment of the invention, the composition comprises active particles comprising heparin, at least 50%, at least 70% or at least 90% of the active particles having a mass median aerodynamic diameter (MMAD) of no more than about 15 μm. Active particles. In another embodiment, at least 50%, at least 70%, or at least 90% of the active particles have an MMAD of from about 10 μm to about 5 μm. In another embodiment, at least 50%, at least 70%, or at least 90% of the active particles have an aerodynamic diameter ranging from about 0.05 μm to about 3 μm. In one embodiment of the invention, at least about 90% of the heparin-containing particles have a particle size (MMAD) of 5 μm or less.

However, particles having a diameter of less than about 10 μm are thermodynamically unstable due to their high surface area to volume ratio, which provides a significant excess of surface free energy and promotes coalescence of the particles. In a dry powder inhaler, the coalescence of small particles and the adhesion of particles to the wall of the inhaler cause the active particles Leaving the inhaler with a large agglomerate, or not leaving the inhaler and still adhering to the inside of the device, or even obstructing or blocking the inhaler.

Uncertainty in the extent to which stable particle aggregates are formed between each start of the inhaler and between different inhalers and between different batches of particles results in poor dose reproducibility. Furthermore, the formation of agglomerates means that the MMAD of the active particles is greatly increased and the active particle agglomerates cannot reach the desired portion of the lungs. Accordingly, it is an important aspect of the present invention to provide a powder formulation that provides good dosage and reproducibility, delivers the correct and predictable dosage.

Much work has been done to improve the efficacy of dry powder systems comprising active granules having a size of less than 15 μm MMAD, reducing the loss of pharmaceutically active agent per delivery stage. In the past, efforts to increase the efficiency of medication and greater reproducibility of medication have been focused on avoiding the formation of agglomerates of fine active agent particles. Such agglomerates increase the effective size of such particles and, therefore, prevent them from reaching the active granules to be deposited to have the desired therapeutic effect of the lower respiratory tract or deep lungs. Proposed means include the use of relatively large carrier particles. The fine active agent particles are susceptible to attachment to the surface of the carrier particles due to intermolecular forces such as Van der Waals forces. When the inhaler device is activated, the active particles should be detached from the carrier particles and then present in the aerosol form in a respirable form. Additionally or alternatively, it has been proposed to include an additive material as a strength control agent which improves the cohesiveness and adhesion between the particles.

However, if the dose of the drug to be delivered is extremely high, the selectivity of the addition of the material to the powder composition is limited, particularly as disclosed in the present invention, wherein at least 90% of the composition is composed of heparin. However, it is essential that the dry powder composition exhibits good flow and dispersion properties to ensure good efficacy. rate.

Heparin is available from a variety of sources, including Bioiberica (such as undifferentiated sodium heparin (Bioiberica, EM/09/140, F0001)).

Another therapeutic agent

According to the invention, heparin can be administered alone (i.e., as a single drug). However, in a further embodiment of the invention, heparin may be administered in combination with an additional therapeutic agent as listed above (e.g., an additional therapeutic agent for the treatment of COPD).

Herein (in another aspect or embodiment of the invention) there is a therapeutic agent (also referred to as an active agent or active ingredient), such as heparin or ADS 115398, which relates to such therapeutic agents themselves, such compounds Tautomers, pharmaceutically acceptable salts or solvates or pharmaceutically functional derivatives of such therapeutic agents.

Pharmaceutically acceptable salts which may be mentioned include acid addition salts and base addition salts. Such salts may be formed by conventional means, for example, by a free acid or free base type of active ingredient (eg, heparin, ADS 115398, etc.) and one or more equivalents of a suitable acid or base, optionally in a solvent. The reaction may be carried out in a medium in which the salt is insoluble, after which the solvent or the medium is removed using standard techniques (for example, by vacuum, by freeze drying or by filtration). Salts can also be prepared by, for example, using a suitable ion exchange resin to exchange opposite ions of a salt form active ingredient (e.g., heparin, ADS 115398, etc.) with another opposite ion.

Examples of pharmaceutically acceptable salts include acid addition salts derived from mineral acids and organic acids, and from metals such as sodium, magnesium or especially potassium and calcium. Salt.

Examples of the acid addition salt include acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, and arylsulfonic acid (for example, benzenesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-di Sulfonic acid and p-toluenesulfonic acid), ascorbic acid (for example, L-ascorbic acid), L-aspartic acid, benzoic acid, 4-ethylguanidinobenzoic acid, butyric acid, (+) sebacic acid. Camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclohexylsulfonic acid, dodecyl sulfate, ethane-1, 2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid, glucoheptonic acid, gluconic acid (for example, D-gluconic acid) , glucuronic acid (for example, D-glucuronic acid), glutamic acid (for example, L-glutamic acid), α-oxoglutaric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid , hydriodic acid, isethionic acid, lactic acid (for example, (+)-L-lactic acid and (±)-DL-lactic acid), lactobionic acid, maleic acid, malic acid (for example, (-)-L- Malic acid), malonic acid, (±)-DL-mandelic acid, metaphosphoric acid, methanesulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, Palmitic acid, phosphoric acid, propionic acid, L-pyroglutamic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, tartaric acid (for example, An acid addition salt formed by (+)-L-tartaric acid, thiocyanic acid, undecylenic acid and valeric acid.

Specific examples of salts are derived from mineral acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, metaphosphoric acid, nitric acid and sulfuric acid; from organic acids such as tartaric acid, acetic acid, citric acid, malic acid, lactic acid, fumaric acid, benzene Formic acid, glycolic acid, gluconic acid, succinic acid, arylsulfonic acid; and salts derived from metals such as sodium, magnesium or especially potassium and calcium.

As mentioned above, the active agents disclosed herein also include active ingredients and the like. Any solvate of the salt. The particular solvate described herein is incorporated by the solid structure (e.g., crystalline structure) of the active agent described herein into a non-toxic pharmaceutically acceptable solvent (hereinafter referred to as a solvent). Solvate formed by the solvent). Examples of such solvents include water, alcohols such as ethanol, isopropanol, and butanol, and dimethyl hydrazine. The solvate can be prepared by recrystallizing the active ingredient in a solvent or a solvent mixture containing a solvating solvent. Whether the solvate is formed in any particular example can be obtained by subjecting the crystallization of the active ingredient to known and standard techniques such as thermogravimetric analysis (TGE), differential scanning calorimetry (DSC), and X-ray crystallography. And decided.

The solvate can be a stoichiometric or non-stoichiometric solvate. Specific solvate hydrates may be mentioned herein, and examples of hydrates include hemihydrate, monohydrate, and dihydrate.

For a hard and detailed discussion of solvates and methods for making and characterizing them, see Bryn et al., Solid-State Chemistry of Drugs, Second Edition, SSCI, Inc of West Lafayette, IN, USA, 1999, ISBN 0-967-06710-3. As defined herein, a "pharmaceutical functional derivative of an active ingredient includes an ester derivative and/or a derivative having or providing the same biological function and/or activity as any of the related compounds of the present invention. Thus, for the purposes of the present invention This term also includes pre-drugs of the active ingredients described herein.

The term "prodrug" of the relevant active ingredient is included in the in vivo metabolism after oral or parenteral administration for a predetermined period of time (eg, within 6 to 24 hours of dosing (eg, one to four times per day)) Any compound that forms an experimentally detectable amount of the compound.

The active ingredient prodrugs described herein can be prepared by modifying the functional groups present on the compound in such a manner that they are modified in vivo to effect cleavage in a mammalian subject. Modification is typically achieved by the synthesis of a parent compound from a previously used drug replacement. Prodrugs include any of the hydroxyl, amine, sulfhydryl, carboxyl or carbonyl groups of the compound of Formula I and any group which can be cleaved in vivo to regenerate free hydroxyl, amine, sulfhydryl, carboxyl or carbonyl groups individually. The active ingredient of the group bond.

Examples of prodrugs include, without limitation, esters of hydroxy functional groups and urethane ester groups, ester groups of carboxy functional groups, N-mercapto derivatives, and N-Mannich bases. General information on prodrugs can be found, for example, in Bundegaard, H. "Design of Prodrugs" p. I-92, Elsevier, New York-Oxford (1985).

As used herein, the term "additional therapeutic agent" includes reference to one or more (e.g., one) therapeutic agents known to be useful in the treatment of COPD (e.g., known to be effective).

As used herein, the phrase "administered sequentially, simultaneously or concomitantly" includes reference to: (i) administration of an individual pharmaceutical formulation (one containing heparin, and one or more other therapeutic agents containing one or more other therapeutic agents) And (ii) administering a single pharmaceutical formulation containing heparin and other therapeutic agents.

In a particular embodiment, heparin and other active agents as defined above are delivered from different containers when heparin is to be administered by pulmonary inhalation.

Other active agents described herein (ie, non-heparin) may be administered by any suitable route, but may in particular be in the form of a pharmaceutically acceptable drug. A pharmaceutical preparation containing the compound by oral, intravenous, intramuscular, dermal, subcutaneous, mucosal (eg, sublingual or buccal), rectal, transdermal, nasal, pulmonary (eg, tracheal or bronchial), topical , by any other parenteral route. Specific modes of administration that may be mentioned include oral, transdermal, intravenous, dermal, subcutaneous, nasal, intramuscular or intraperitoneal administration. More specific modes of administration that may be mentioned include oral and transdermal administration.

The other active agents described herein will generally be administered in a pharmaceutical formulation in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier which may be selected according to the desired route of administration and standard pharmaceutical practice. Such pharmaceutically acceptable carriers can be chemically inert to the active compound and have no adverse side effects or toxicity under the conditions of use. Suitable pharmaceutical formulations are found, for example, in Remington The Science and Practice of Pharmacy, 19th edition, Mack Printing Company, Easton, Pennsylvania (1995). For parenteral administration, parenterally acceptable aqueous solutions can be employed which are pyrogen free and have the necessary pH, isotonicity, and stability. Suitable solutions are well known to those skilled in the art, and several methods are described in the literature. A brief review of drug delivery methods can also be found, for example, in Langer, Science (1990) 249, 1527.

Otherwise, the preparation of a suitable formulation can be conventionally accomplished by those skilled in the art using conventional techniques and/or in accordance with standard and/or accepted pharmaceutical practice.

The amount of other active agents described herein in any pharmaceutical formulation for use in accordance with the present invention will depend on a variety of factors, such as the severity of the condition to be treated, the particular patient to be treated, and the compound employed. anyway, The amount of other active agents described herein within the formulation may be conventionally determined by those skilled in the art.

For example, a solid oral composition such as a lozenge or capsule may contain from 1 to 99% (w/w) active ingredient; from 0 to 99% (w/w) diluent or filler; from 0 to 20% ( w/w) disintegrant; from 0 to 5% (w/w) lubricant; from 0 to 5% (w/w) of flow aid; from 0 to 50% (w/w) a chemical or binder; from 0 to 5% (w/w) of an antioxidant; and from 0 to 5% (w/w) of a colorant. The controlled release lozenge may additionally contain from 0 to 90% (w/w) of the release controlling polymer.

Parenteral formulations (such as solutions or suspensions for injection, or solutions for infusion) may contain from 1 to 50% (w/w) of active ingredient; and from 50% (w/w) to 99% (w/w) liquid or semi-solid carrier or vehicle (for example, a solvent such as water); and 0-20% (w/w) one or more other excipients, such as buffers, antioxidants , suspension stabilizer, osmoticity adjuster, and preservatives.

Depending on the abnormality to be treated and the patient's route of administration, the other active agents described herein can be administered to patients in need thereof at different therapeutically effective doses.

However, in the context of the present invention, the dosage administered to a mammal, particularly a human, is sufficient to produce a therapeutic response in a mammal for a reasonable period of time. Those skilled in the art will understand that the correct dosage and composition selection and the most suitable delivery method are also subject to the pharmacological properties of the formulation, the nature and severity of the condition to be treated, and the physical and mental acuity of the recipient, with the particular compound. Effectiveness, age, condition, weight, sex and/or the patient to be treated The response, and the stage/severity of the disease, etc.

Administration can be continuous or intermittent (eg, by bolus injection). The dose can also be determined by the time and frequency of administration. In the case of oral or parenteral administration, the dosage may vary from about 0.01 mg to about 2000 mg of the other active agent described herein (eg, 0.5 mg, 3 mg, 6 mg or 10 mg of ADS 115398 per day). .

In any event, the physician or other skilled artisan can routinely determine the actual dosage that is most suitable for the individual patient. The above dosages are exemplified by the average case; it is of course possible to have individual cases requiring higher or lower dosage ranges, which are within the scope of the invention.

The various aspects of the invention described herein (e.g., the compounds, compositions, methods, and uses described above) can have the advantage that treatment in the above-described conditions is known to those skilled in the art for treating such conditions or other It is more convenient, more effective, less toxic, more selective, and has a wider range of activity, and is more convenient for physicians and/or patients than similar compounds, compositions, methods (treatments) or uses. Effective, produces fewer side effects, or may have other useful pharmacological properties.

additive

The tendency of fine particles to coalesce means that a particular dose of FPF will be highly unpredictable and that different ratios of fine particles will therefore be applied to the lungs or to the correct part of the lungs. This is observed, for example, in a formulation containing a pure drug in the form of a fine particle. These formulations exhibit poor flow properties and poor FPF.

In order to improve this situation and provide consistent FPF and FPD efforts, based on this issue The dry powder composition of the present invention may include a material which is an anti-adhesive material and an additive material which reduces the cohesiveness between the particles in the composition.

The additive material is selected to reduce cohesion between the particles within the dry powder composition. The additive material is believed to interfere with the weak bonding forces between the small particles, helping to keep the particles separate, and to reduce the adhesion of the particles to each other, to the other particles of the formulation, if present, and to the inner surface of the inhaler device. If agglomerates of particles are formed, the addition of the agglomerates of the additive material particles is such that they are more likely to rupture in the turbulent gas stream generated by the activation of the inhaler device, so that the particles are discharged from the device and are inhaled. . When the agglomerates rupture, the active particles can return to a pattern of small individual particles or agglomerates of small numbers of particles that can reach the lower lung.

As disclosed in WO 1997/03649, the additive material can be in the form of particles that tend to adhere to the surface of the active particles. Additionally, the additive material can be applied to the surface of the active particles, for example, by the co-milling process as disclosed in WO 2002/43701.

In particular, the additive material is an anti-adhesive material and it tends to reduce cohesion between the particles and also prevents fine particles from adhering to the surface within the inhaler device. Advantageously, the additive material is an anti-friction or glidant and imparts better flow properties to the powder formulation in the inhaler. The additive materials used in this manner may not necessarily be referred to as anti-adhesives or anti-friction agents, but they may have the effect of reducing inter-particle cohesion or improving powder flow. Additive materials are sometimes referred to as force control agents (FCAs), and their like generally result in better dose reproducibility and higher FPF.

Therefore, the additive material or FCA is used in other particles when used here. A material that is present and associated with the surface of the particle that is present on the surface of the particle that alters the adhesion of the particle and the strength of the cohesive surface. In general, its function is to reduce adhesion and cohesion.

The tendency to reduce the strong binding of the particles to each other or to the device itself not only reduces powder cohesion and adhesion, but also promotes better flow characteristics. This results in an improvement in dose reproducibility as it reduces the amount of powder metered out per dose and improves the release of the powder from the device. This also increases the likelihood that the active material leaving the device will reach the lower lung of the patient.

When present in an inhaler device, it is advantageous for the unstable particulate agglomerates to be present in the powder system. As indicated above, in order for the powder to leave the inhaler device efficiently and reproducibly, the coalesced particles of the powder need to be large, in particular greater than about 30 μm, preferably greater than 50 μm, and more preferably greater than 70 μm. It is optically measured using, for example, a scanning electron microscope and a suitable reference sample of known size. The powder may be in the form of individual particles having a size of about 40 μm or more, and/or agglomerates of finer particles having a size of about 40 μm or more. The agglomerates formed may have a size of 100 μm or 200 μm, and depending on the type of device used to dispense the formulation, the agglomerates may be equal to about 1000 μm. These agglomerates are more likely to effectively rupture within the turbulent gas stream produced by the inhalation by the addition of additive materials. Thus, it may be advantageous to form unstable or "soft" particulate agglomerates within the powder as compared to particles that do not substantially have coalescence. This unstable agglomerate is stable when the powder is not inside the device, but disperses and ruptures upon inhalation.

It is particularly advantageous if the additive material comprises an amino acid. It has been found that when an amino acid is present as an additive material, an active material having a high respirable fraction is produced, And the good flow properties of the powder. A particular amino acid leucine, such as L-leucine, di-leucine, and tri-leucine, may be mentioned. Although the L-type amino acid is usually used, the D- and DL-forms can also be used. The additive material may comprise one or more of any of the following amino acids: aspartame, leucine, isoleucine, lysine, valine, methionine, cysteine, and phenylpropylamine. acid. Additive materials may also include, for example, metal stearates such as magnesium stearate, phospholipids, lecithin, colloidal cerium oxide, and sodium stearyl fumarate, and are more fully described in WO 1996/23485, It is hereby incorporated by reference.

Advantageously, the powder comprises at least 80%, at least 90%, or especially 95% (eg, at least 96%, 97%, 8% or 99%) by weight of the glycosaminoglycan (eg, heparin) based on the weight of the powder. ) optionally containing other active ingredients such as those discussed herein. The optimum amount of additive material will depend on the precise nature of the additive and the manner in which it is incorporated within the composition. In certain embodiments, the powder advantageously comprises no more than 8%, more preferably no more than 6%, preferably 5% by weight of additive material, based on the weight of the powder. As indicated above, in some cases it is advantageous for the powder to contain about 1% by weight of additive material. In other embodiments, the additive material or FCA can be provided in an amount from about 0.1% to about 10% by weight, and especially from about 0.15% to 7%, most specifically from about 0.5% to about 6%.

When the additive material is micronized leucine or lecithin, it is from about 0.1% to about 10% by weight (eg, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%). The amount of 7%, 8% or 9%) is specifically provided. In particular, the additive material comprises from about 3% to about 7%, especially about 5%, of micronized amine acid. In particular, at least 95% (eg at least 96%, 97%, 98% or 99%) by weight of micronized leucine has less than 150 μm, in particular less than 100 μm, and most particularly less than 50 μm Particle diameter. In particular, the mass median diameter of the micronized leucine is less than 10 μm.

If magnesium stearate or sodium stearyl fumarate is used as the additive material, depending on the desired final dosage, it is from about 0.05% to about 10%, from about 0.15% to about 5%, and from about 0.25% to about Approximately 3%, or from about 0.5% to about 2.0%, is specifically provided.

In an effort to improve the extraction of the dry powder self-dispensing device and to provide consistent FPF and FPD, the dry powder composition according to the present invention may comprise particles of an inert excipient material as carrier particles. These carrier particles are mixed with the fine active material particles and any additive materials present. When in the inhaler device, they do not adhere to each other. The fine active particles adhere to the surface of the carrier particles, but should be released and dispersed when the dispensing device is activated, and sucked into the respiratory tract to produce fine suspension.

The inclusion of carrier particles is less attractive when a large dose of active agent is to be delivered because it tends to significantly increase the volume of the powder composition. However, in certain embodiments of the invention, the composition comprises carrier particles. In one embodiment, the composition comprises at least about 10% by weight of active ingredient (eg, heparin, alone or in combination with one or more active ingredients), or at least about 15%, 17%, or 18 % or 18.5% by weight of active ingredient (for example, heparin, alone or in combination with one or more active ingredients). More particularly, the carrier particles are present in minor amounts, such as no more than 90% (e.g., 85%, 83% or more particularly 80%) by weight of the total composition, wherein the total active ingredient (e.g., liver) The content of the prime and magnesium stearate is about 18.5 and 1.5% by weight, respectively.

The carrier particles can be any acceptable inert excipient material or combination of materials. For example, the carrier particles can comprise one or more materials selected from the group consisting of sugar alcohols, polyols, and crystalline sugars. Other suitable carriers include inorganic salts such as sodium chloride and calcium carbonate, organic salts such as sodium lactate, and other organic compounds such as polysaccharides and oligosaccharides. Advantageously, the carrier particles comprise a polyol. In particular, the carrier particles can be particles of crystalline sugar, for example, mannitol, trehalose, melezitose, dextrose, or lactose. Most particularly, the carrier particles are composed of lactose.

Thus, in one embodiment of the invention, the composition comprises active fruit granules (including heparin) and carrier particles. The carrier particles can have an average particle size of from about 5 to about 1000 μm, from about 4 to about 40 μm, from about 60 to about 200 μm, or from 150 to about 1000 μm. Other useful average particle sizes of the carrier particles are from about 20 to about 30 μm, or from about 40 to about 70 μm.

In another embodiment, the carrier particles are present in minor amounts, such as no more than 50% by weight of the total composition (eg, 60%, 70%, or more specifically 80%), wherein the total active ingredient ( For example, heparin) and magnesium stearate (weight) are 18 and 2% by weight. The amount of additive and heparin will also vary as the loading dose within such formulations changes, but the ratio of such components is specifically maintained from about 1:9 to about 1:13.

In another embodiment, the formulation contains no carrier particles and comprises an active ingredient (eg, heparin) and an additive, such as at least 30% by weight of the total composition (eg, 60%, 80%, 90%, 95%, Or more particularly 97%) comprises a pharmaceutically active agent. The active agent can be a single glycosaminoglycan (eg, heparin), Or it may be a combination of a glycosaminoglycan (eg, heparin) and another drug that is beneficial to a patient (eg, a patient with COPD). The remaining components may comprise one or more additive materials such as those discussed herein.

In another embodiment, the formulation may contain carrier particles and comprise an active ingredient (eg, heparin alone or in combination with one or more active ingredients) and an additive, such as at least 30% by weight of the total composition (eg, 60%, 80%, 90%, 95%, or more particularly 97%) comprises a pharmaceutically active agent, and wherein the remaining components comprise the additive material and larger particles. Larger particles provide a dual role as a carrier and promote powder flow.

In a particular embodiment, the composition comprises heparin (30% w/w) and lactose having an average particle size of 45-65 μm. In a particular embodiment, the composition comprises heparin (90% w/w, for example, 95%, such as 96%, 97%, 98%, or 99%) and lactose having an average particle size of 45-65 μm.

The composition comprising the active ingredient (for example, heparin alone or in combination with one or more active ingredients) and the carrier particles may further comprise one or more additive materials. The additive material can be in the form of particles that tend to adhere to the surface of the active particles, as disclosed in WO 1997/03649. Alternatively, the additive material can be applied, for example, by co-milling methods to the surface of the active particles (as disclosed in WO 2002/43701) or to the surface of the carrier particles (as disclosed in WO 2002/00197).

In one embodiment, the additive is applied to the surface of the carrier particles. Because the carrier is admixed with the additive, the coating can be in the form of particles of additive material (by intergranular forces such as van der Waals) that adhere to the surface of the carrier particles. In addition, the additive material is applied and fused to the surface of the carrier particles, whereby A composite particle having an inert carrier material core and an additive material on the surface is formed. For example, the fusion of the additive material with the carrier particles can be achieved by co-flow milling the additive material particles and the carrier particles. In certain embodiments, all three components of the powder (active agent, carrier, and additive) are compressed together such that the additive adheres or fuses to the carrier particles and the active agent particles s. In an exemplary embodiment, the composition comprises an additive material, such as magnesium stearate (up to 10% w/w) or leucine, which is ground with heparin particles and/or with a stream of lactose.

In one particular embodiment described herein, the formulation comprises one or more of the following: (a) an additive material (eg, magnesium stearate); and (b) a carrier (eg, lactose fines).

In certain embodiments of the invention, the heparin formulation is a "carrierless" formulation comprising only the active ingredient (eg, heparin) or a pharmaceutically acceptable salt or ester thereof and one or more additive materials.

In the preferred embodiment described herein, the formulation comprises an additive material, preferably wherein the additive material is an amino acid, more preferably, the amino acid is L-leucine.

Advantageously, in such "unloaded" formulations, at least 90% by weight of the powder particles have a particle size of less than 63 μm, in particular less than 30 μm and more particularly less than 10 μm. As indicated above, in order to be effectively delivered to the lower lung, the size of the particles of powdered heparin (or a pharmaceutically acceptable salt thereof) needs to be in the range of about 0.1 μm to 5 μm. If the additive material is in the form of a granule, for delivery to the lower lung, it may be advantageous for such additives to have this preferred range. size.

The powder comprises at least 60% by weight of a glycosaminoglycan (e.g., heparin) or a pharmaceutically acceptable salt or ester thereof, based on the weight of the powder. Advantageously, the powder comprises at least 70%, or at least 80% by weight, of a glycosaminoglycan (eg, heparin) or a pharmaceutically acceptable salt or ester thereof, based on the weight of the powder. Most advantageously, the powder comprises at least 90%, at least 95% (eg, at least 96%), or at least 97% (eg, at least 98% or 99%) by weight of heparin or a pharmaceutically acceptable amount thereof, based on the weight of the powder. Salt or ester. It is believed that the material which is physiologically beneficial to the lungs as small as possible, especially the inactive ingredients, is administered to the patient. Therefore, the amount of additive material added is particularly as small as possible. Thus, most particularly, the powder will comprise more than 99% by weight of heparin or a pharmaceutically acceptable salt or ester thereof.

In a particular embodiment, it is determined by X-ray powder diffraction (XRPD) analysis that at least some of the heparin is in an amorphous form. Formulations containing amorphous heparin will have special solubility characteristics. Stable forms of amorphous heparin can be prepared using suitable sugars such as trehalose and melezitose.

Preparation of dry powder inhaler formula

If the composition of the present invention includes an additive material, the manner in which it is incorporated has a significant impact on the effect of the additive material on the properties of the powder, including FPF and FPD.

In one embodiment, the composition according to the present invention is obtained by granules and additive material particles and/or carriers having active ingredients selected to be appropriately sized (eg, heparin alone or in combination with one or more active ingredients). The particles are prepared by simply blending. The powder component can be blended by a mild mixing method For example, in a drum mixer, such as Turbula (trademark). In this mild mixing process, the size of the particles to be mixed is generally not substantially reduced. In addition, the powder particles are not easily fused to each other, but they are agglomerated by cohesion such as the strength of the house. Such relaxed or unstable agglomerates are easily broken when the inhaler device for dispersing the composition of the present invention is activated.

Compression grinding method

In another aspect for preparing a composition according to the present invention, the powder component is subjected to a compression milling method such as a method called mechanical fusion (also referred to as 'mechanical chemical bonding') and rotary mixing.

As the name implies, mechanical fusion is a dry coating process designed to mechanically fuse a first material to a second material. It should be noted that the use of "mechanical fusion" and "mechanical fusion" should be interpreted as relating to a particular type of grinding method, but not to a grinding method implemented in a particular apparatus. The compression grinding process is based on different principles of other grinding techniques, depending on the particular interaction between the internal components and the walls of the vessel, and is based on the energy provided by the controlled and substantial compression forces. This method works particularly well when a material is generally smaller and/or softer than others.

The fine active particles and the additive particles are supplied to a mechanical fusion device such as a container of a Mechano-Fusion system (Hosokawa Micron Ltd) or a Nobilta or Nanoocular device, in which the cells are subjected to centrifugal force and pressed against the inner wall of the container to suppress the powder. Compressing between the drum wall and the fixed gap of the inner member of the inner portion at a high relative speed between the drum and the member. The inner wall and the deflecting member generally form a gap or an angle, and the particles are pressed therebetween. Together. Thus, when limited between the inner drum wall and the inner member (which has a greater curvature than the inner drum wall), the particles encounter extremely high shear forces and extremely strong compressive stresses. The particles are pressed against each other and with sufficient energy to locally heat and soften, rupture, distort, flatten, and surround the core particles to form a coating. This energy is generally sufficient to cause the agglomerates to rupture and a certain degree of size reduction of the two components will occur.

These mechanical fusion and rotary mixing methods exert a sufficiently high degree of force to separate individual active material particles and to rupture the compacted active particle agglomerates such that effective mixing of the additive materials and effective application to the surface of such particles is achieved. One of the special aspects of this method is that the additive material is deformed during grinding and will be applied or fused to the surface of the active particles.

However, in practice, such compression milling methods produce little or no reduction in drug particle size, especially when they have been micronized (i.e., < 10 μm). The only observable physical change is the plasticity of the particles forming a rounded shape.

Other grinding procedures

This method of milling can also be used to formulate dry powder compositions in accordance with the present invention. The production of fine particles by grinding can be achieved using conventional techniques. When used conventionally, "grinding" means applying any mechanical force to the active material particles using any mechanical means to break coarse particles (eg, particles having MMAD greater than 100 μm) into fine particles (eg, , with MMAD of no more than 50 μm). In the present invention, the term "grinding" also refers to the deagglomeration of particles within a formulation with or without a reduction in particle size. The particles to be ground may be large or fine before the grinding step. Wide range of grinding devices and strips The articles are suitable for use in making the compositions of the present invention. The choice of suitable milling conditions to provide the desired level of strength, such as abrasive strength and time, will be within the skill of the artisan.

The impact milling method can be used to prepare a heparin-containing composition according to the present invention with or without an additive material. This method includes ball milling and the use of a mixer.

Ball milling is one of the common grinding methods used in the art for polishing methods. Centrifugal and row ball milling are particularly preferred methods.

Additionally, a high pressure mixer can be used in which a flow system containing particles is forced through a valve at high pressures that produce high shear and turbulent conditions. Shear forces on the particles, impacts between the particles and mechanical surfaces or other particles, and cavitation due to fluid acceleration can contribute to particle splitting. Suitable mixers include EmulsiFlex high pressure mixers with pressures up to 4000 bar, Niro Soavi high pressure mixers (with pressures up to 2000 bar), and Microfluidics Microfluidisers (maximum pressure 2750 bar). The milling method can be used to provide microparticles having a mass median aerodynamic diameter as specified above. The mixer can be more suitable for large-scale preparation of composite active particles than a ball mill.

The grinding step may additionally comprise a high energy bead mill or agitator bead mill, for example a Netzsch high energy bead mill, or a DYNO-grinder (Willy A. Bachofen AG, Switzerland).

A common airflow milling system as disclosed in the earlier patent application published in WO2005/025536 is specifically used if it is also desired to reduce the particle size. Common airflow grinding methods result in low or submicron straight Composite active particles of the diameter, and these particles exhibit particularly good FPF and FPD, even when using passive DPI dispensing.

The grinding method applies a sufficient degree of force to break the tightly bound fine or ultrafine particle agglomerates such that effective mixing of the additive materials and effective application to the surface of the particles is achieved.

These impact methods produce high energy impacts between the beads and between the particles or between the particles. In fact, although these methods are good at making very small particles, it has been found that neither the ball mill nor the mixer has been particularly effective in producing a dispersion of the drug powder in a manner that is observed by the compression method. It is believed that the second impact method does not effectively produce a coating of additive material on each particle.

A conventional method comprising co-milling an active material with an additive material (as described in WO 2002/43701) forms a composite active particle which is a fine active material particle having a content of additive material on its surface. The additive material is in particular a coated form on the surface of the active material particles. This coating can be a discontinuous coating. The additive material can be in the form of particles that adhere to the surface of the active material particles. Co-milling or co-micronization of the particles of the active agent particles and additives (FCA) or excipients will form an additive or excipient that is deformed and coated or fused to the surface of the active particles composed of the two materials. These formed composite active particles comprising the additive were found to be less cohesive after the grinding treatment.

At least some of the composite active particles may be in the form of agglomerates. However, when the composite active particles are included in the pharmaceutical composition, the additive material promotes dispersion of the composite active particles upon administration of the composition to the patient via a start-up inhaler.

Grinding can also be used in the presence of materials that delay or control the release of the active agent. Row.

Co-milling or co-micronization of the active agent and additive particles can include methods of compression, such as mechanical fusion, rotary mixing, and related methods such as those involving the use of Hybridiser or Nobilta. The principles behind these methods are different from those of other grinding techniques because they involve a special interaction between the internal component and the wall of the vessel, and because it provides energy by the force of controlled and solid compression. The foundation, in particular the compression within the gap of the predetermined width.

In one embodiment, the microparticles produced by the milling step can then be formulated with additional excipients, if desired. This can be achieved by a spray drying process, for example, spray drying with an excipient. In this embodiment, the granules are suspended in a solvent and spray dried together with a solution or suspension of additional excipients. In particular additional excipients include trehalose, melezitose and other polysaccharides. Further pharmaceutically effective excipients can also be used.

In another embodiment, the powder composition is made using a multi-step process. First, the material is ground or blended. Second, the particles can be screened prior to mechanical fusion. A further optional step involves the addition of carrier particles. The mechanical fusion step is believed to "polish" the composite active particles, further grinding the additive material into the active particles. This allows for the advantageous properties provided to the particles by mechanical fusion and in combination with the extremely small particle size that may be produced by gas flow milling.

The cohesion and adhesion reduction between the active particles can result in properties that are equivalent to reducing the size of the agglomerates or even individual particles.

High shear blending

Expanding pharmaceutical product manufacturing specifications typically requires the use of one piece of equipment to perform more than one function. An example of this is the use of a mixing granulator which mixes and granulates the product, thereby eliminating the need to transfer the product between the items of the equipment. In this way, the chance of powder isolation is minimal. High shear blending typically uses a high shear rotor/stator mixer (HSM) that has been used in hybrid applications. The mixer or "high-cut material processor" develops high pressure on the material whereby the mixture is then transported through a very fine orifice or at an acute angle. Depending on the material to be processed, the flow through the chamber may be countercurrent or cocurrent. The number of chambers can be increased to achieve better performance. The orifice size or impact angle can also be varied to optimize the particle size produced. The reduction in particle size occurs due to the high shear produced by the high shear material processor as it passes through the orifice and chamber. The ability to apply strong shear and shorten the mixing cycle makes these mixers attractive for applications where uniform blending of the coalesced powder is desired. Furthermore, conventional HSM can also be widely used for high-intensity mixing, dispersion, disintegration, emulsification, and homogenization.

It is known to those skilled in the art to make small particle powder formulations, even with high power, high shear mixing machines, which require a relatively long "aging" period to achieve complete dispersion, and this period cannot be increased by mixing power. Or increase the speed of the mixer to increase the shear rate and slightly shorten. A high shear mixer can also be used if the self-adhesive nature of the drug particles is such that high shear forces are required to form a particle coating or film that reduces surface energy with a strength control agent.

Spray drying and ultrasonic atomizer

Spray drying can be used to make particles containing the respirable size of heparin. Spray drying method can be used to manufacture active agents and control particle coalescence and powder Spray dried particles of the final performance additive material. Spray drying methods can also be used to make spray dried granules comprising an active agent dispersed or suspended in a material that provides controlled release properties. Furthermore, dispersion or suspension of the active material within the excipient material can impart further stability to the active compound. In a particular embodiment, heparin can be present primarily in an amorphous state. Formulations containing amorphous heparin will have special solubility characteristics. This is possible because the particles are suspended in a sugar glass which can be a solid solution or a solid suspension. In particular additional excipients include trehalose, melezitose and other polysaccharides.

Spray drying is a technique known and widely used for making respirable sized active material particles. Conventional spray drying techniques can be modified to produce active particles with promoted chemical and physical properties that are better implemented when dispensed from DPI than particles formed using conventional spray drying techniques. Such improvements are described in detail in earlier patent applications published as WO 2005/025535.

In particular, it has been disclosed that co-spray drying of an active agent with FCA under special conditions can form particles having excellent properties which are particularly well carried out when inhaled into the lungs by DPI administration.

It has been found that manipulating or adjusting the spray drying process can result in a substantial amount of FCA being present on the surface of the particles. That is, the FCA is concentrated, but non-exclusively located on the surface of the particles as individual crystals, and is uniformly distributed throughout the particles. This clearly means that FCA will reduce the tendency of particles to coalesce. This can help to form unstable agglomerates that rupture easily and consistently when DPI is activated.

Advantageously, when measured by X-ray powder diffraction (XRPD) dispersion or atomic force microscopy or a combination thereof, the particle system comprises, based on the weight of the powder At least 2%, at least 3%, or especially 4% (by weight of the particles) of the additive on the surface of the granules, optionally comprising the active ingredient. The optimum amount of additive material will depend on the precise nature of the additive and the manner in which it is included in the composition.

It has been found that it may be advantageous to control the formation of drops of the spray drying method such that drops of a particular size and narrow size distribution are formed. Further, controlling the formation of the drip can control the flow of air around the drip, and thus can be used to control the drying of the drip, and particularly the drying rate. Controlling droplet formation can be achieved by using alternatives to conventional 2-fluid nozzles, particularly to avoid the use of high rate gas streams.

In particular, it is preferred to use a spray dryer comprising one of the means for producing a drip that moves at a controlled rate and has a particular drip gauge. The drip rate is specifically controlled relative to the gas body into which it is sprayed. This can be achieved by controlling the rate of drop initiation and/or the rate at which it is sprayed into the body of the gas, for example, by using an ultrasonic atomizer (USN). Alternative nozzles such as electrospray or vibrating orifice nozzles can be used.

In one embodiment, USN is used to form a drop of spray. The USN uses an ultrasonic transducer that is submerged in the liquid. Ultrasonic transducers (piezoelectric crystals) vibrate at ultrasonic frequencies, producing the short wavelengths required for liquid atomization. In a general version of USN, the crystalline bottom is fixed such that the vibrational system is transferred from its surface to the nebulizer liquid either directly or via a coupling liquid (typically water). When the ultrasonic vibration is sufficiently strong, a liquid fountain is formed on the surface of the liquid in the atomizer chamber. The drip is emitted from the apex and emits a "fog".

Although USN is known, these are conventionally used in inhaler devices for direct inhalation of solutions containing drugs, and which have not previously been widely used in spray drying devices. Found that this atomizer is used for spray drying with several weights There are advantages, and these are not previously recognized. The particular USN controls the particle rate and, therefore, the rate at which the particles are dried, thereby affecting the shape and density of the formed particles. The use of USN also provides the opportunity to perform spray drying on a larger scale than would be possible with conventional spray drying devices having conventional type nozzles (e.g., 2-fluid nozzles) for producing drip.

The attractive properties of USN to produce fine particle dry powder include: low spray rate; small amount of carrier gas required to operate the atomizer; smaller droplet size and narrow droplet size distribution; USN Simple nature (no moving parts, it will wear, pollution, etc.); accurately control the airflow around the drip, thereby controlling the drying rate; and high output rate, which can use the commercially available USN (which uses the flow of the second Fluid nozzles are configured to produce dry powder in a difficult and expensive manner.

USN does not separate the liquid into drops by increasing the rate of liquid. Conversely, the required energy is provided by the vibrations caused by the ultrasonic atomizer.

Further embodiments may utilize the use of an ultrasonic atomizer (USN), a rotary atomizer, or an electrohydrodynamic (EHD) atomizer to produce particles.

Delivery device

The inhalable composition according to the invention is administered in particular via a dry powder inhaler (DPI), but can also be administered via a pressurized metered dose inhaler (pMDI), via a liquid drip, or even via an atomization system . In one embodiment of the invention, the pulmonary inhalation system is inhaled by an inhaler (eg, DPI or pMDI) or via an atomization system.

In a dry powder inhaler, the dose to be administered is stored in a non-pressurized dry powder form, and when the inhaler is activated, the powder particles are The finely dispersed particle mist pattern inhaled by the patient is discharged from the device.

A dry powder inhaler can be a "passive" device in which the patient's respiratory system provides the sole source of gas for moving forces within the device. Examples of "passive" dry powder inhalers include those manufactured by Vectura (covered by International Patent Application No. WO 2010/086285), or in particular Rotahaler and Diskhaler (GlaxoSmithKline), Monohaler (MIAT), GyroHaler (trademark) (Vectura), Turbohaler (Astra-Draco) and Novolizer (trademark) (Viatris GmbH). Additionally, an "active" device may be used in which a source of compressed gas or another source of energy is used. Examples of suitable active devices include Aspirair (trademark) (Vectura) and active inhaler devices manufactured by Nektar Therapeutics (covered by U.S. Patent No. 6,257,233).

It is generally believed that different compositions are implemented differently when using passive and active inhaler dispensing. The passive device produces less turbulence within the device and the powder particles move more cushioned as they exit the device. This results in some quantitative doses remaining in the device and depending on the nature of the composition, less deagglomeration at startup. However, when the slow moving force mist is inhaled, it is generally observed that the throat is less deposited. Conversely, active devices generate more turbulence at startup. This results in more quantitative doses being extracted from the bubbles or capsules and more deagglomerated because the powder accepts greater shear forces. However, the particle system moves away from the device faster than with a passive device, which can result in increased throat deposition.

It has been surprisingly found that the compositions of the invention having a high ratio of heparin are well implemented when dispensed using active and passive devices. Although the different types of inhaler devices are expected to have some losses everywhere, as described above, this loss The system is extremely small and still allows a large ratio of quantitative doses of heparin to be deposited in the lungs. Once in the lungs, heparin is rapidly absorbed and exhibits excellent bioavailability.

In particular, the "active" dry powder inhaler, which may be referred to herein, refers to an Aspirair® inhaler and is described in more detail in WO 2001/00262, WO 2002/07805, WO 2002/89880 and WO 2002/89881. The contents of the description are incorporated herein by reference. A particular "passive" dry powder inhaler, which may be referred to herein, is described in the "passive" dry powder inhalation device of WO 2010/086285. However, it is to be understood that the compositions of the present invention can be administered in a passive or active inhaler device.

In another embodiment, the composition is a solution or suspension that is dispensed using a pressurized metered dose inhaler (pMDI). The composition according to the present invention may comprise a dry powder composition as discussed above mixed or dissolved with a liquid propellant such as HFA 134a or HFA 227. In another embodiment, the composition is a solution or suspension and is administered using a pressurized metered dose inhaler (pMDI), an atomizer, or a soft atomizer inhaler. Examples of suitable devices include pMDI such as Modulite® (Chiesi), SkyeFineTM and SkyeDryTM (SkyePharma), atomizers such as Porta-Neb®, InquanebTM (Pari) and AquilonTM, and such as eFlowTM (Pari), AerodoseTM (Aerogen), Soft nebulizer inhalers for Respimat® inhalers (Boehringer Ingelheim GmbH), AERx® inhalers (Aradigm) and MysticTM (Ventaira Pharmaceuticals, Inc.).

If the composition is to be dispensed using pMDI, the composition comprising heparin optionally further comprises a propellant (ie, further comprising a propellant). In the present invention In an embodiment, the propellant is CFC-12 or a non-CFC propellant that is ozone friendly, such as 1,1,1,2-tetrafluoroethane (HFC-134a), 1, 1, 1, 2, 3, 3,3-heptafluoropropane (HFC-227), HCFC-22 (difluorochloromethane), HFA-152 (difluoroethane and isobutylene), or combinations thereof. Such formulations may need to include polar surfactants such as polyethylene glycol, diethylene glycol monoethyl ether, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, propoxyl Polyethylene glycol, and polyoxyethylene lauryl ether, which are used to suspend, dissolve, humidify, and emulsify active agents and/or other components, and valve assemblies for lubricating MDI.

It is to be understood that the particular embodiments described herein are shown by way of illustration and not limitation. The main features of the present invention are used in various embodiments without departing from the scope of the invention. Those skilled in the art will recognize or be able to ascertain using only a few routine studies of the particular procedures described herein. Such compounds are considered to be within the scope of the invention and are covered by the scope of the patent application. All publications and patent applications mentioned in the specification are indicated by the extent of those skilled in the art. All publications and patent applications are hereby incorporated by reference to the extent of the same extent as the disclosures The use of the words "a" or "an" may mean "one", but also "one or more", "at least" A ", and "one or more than one" meaning is the same. The use of the word "or" in the context of the patent application is intended to mean "and/or" unless expressly indicated that only one of the alternatives or the alternatives are mutually exclusive, even if the disclosure refers to only one of the The definition of "and / or". In the entire application, the word "about" It is used to refer to the inherent variation of the device, the method used to determine the value, or the value of the variation in the subject.

As used in the specification and claims, "comprising" (and any type of inclusion, such as "comprise" and "comprises"), "having" (and any type of possession, such as, "have" "and" has"), "including" (and any type includes, such as "includes" and "include") or "containing" (and any type of content, such as "contains" and The word "contain" is used inclusive or open-ended and does not exclude additional elements or method steps not described.

The use of the term "or a combination of the same" as used herein refers to all the permutations and combinations of items listed herein. For example, "A, B, C, or a combination thereof" is intended to include at least one of the following: A, B, C, AB, AC, BC, or ABC, and if the order is important in a particular case, BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing with this example, it explicitly includes repeaters that contain one or more items or words, such as BB, AAA, MB, BBC, AAABCCCC, CBBAAA, CABABB, and the like. Those skilled in the art will appreciate that there is typically no limit to the number of items or words used in any combination, unless explicitly different from the circumstances.

All of the compositions and/or methods disclosed and claimed herein can be implemented and executed without undue experimentation based on the disclosure. Although the compositions and methods of the present invention have been described in terms of specific embodiments, it will be apparent to those skilled in the art that Or the method and the steps of the method or the sequence of the steps are changed. Replacement and repair of all such similarities apparent to those skilled in the art The modifications are considered to be within the spirit, scope and concept of the invention as defined in the appended claims.

example Example 1

Undifferentiated heparin sodium (100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg) was administered intranasally (in) to evaluate lungs induced by exposure to tobacco smoke (TS) in C57BL/6J mice for 4 days. The effect of inflammation. Inhaled p38 MAP kinase inhibitor, ADS115398, was administered as a reference agent at 100 μg/kg i.n. 1 hour before each TS exposure.

Experimental procedure

The total study period was approximately 5 days from the start of the study and samples were taken from the last subject. The evaluable subjects were randomly assigned to the treatment group.

Carrier for intranasal administration

0.2% Tween 80 (Product No.: C-4888, Lot No. 82K0153).

Test substance

Undifferentiated sodium heparin (Bioiberica, EM/09/140, F0001) was pre-weighed in a vial ready to be used for daily administration of new preparations (agreeed amount).

Reference control group

ADS115398 (p38 MAP kinase inhibitor)

animal

Seventy (10 in each group) female mice were 16-20 g (C57BL/6J), were housed under a complete barrier and were confirmed to have no specific microorganisms upon receipt. Individual Pentel marks on the tail are used as authentication methods.

therapy group

Number of groups: 7; size of the group: 10; dose volume: 50 μL per mouse (25 μL per nostril); treatment time; intranasal, once daily

Bronchoalveolar perfusion (BAL) was performed using 0.4 ml of phosphate buffered saline (PBS). Cells recovered from BAL were used for total cell and differential cell counts performed on slides prepared using a cytospin. The BAL supernatant was stored at -40 °C.

data analysis

Normality testing is performed on cellular data. The primary analysis was then performed using the single factor analysis test (single factor ANOVA) followed by the Bonferroni multiple comparison test to compare the control and treatment groups and/or the Mann Whitney test. Unusually distributed data is used The Kruskal-Wallis test and subsequent Dunn multiple comparison tests were evaluated. Data p < 0.05 was considered statistically significant.

result

There was a statistically significant (P = < 0.01) decrease in total cell counts in all heparin treated groups. This appears to be the dose associated with the two doses (1000 μg/kg and 2000 μg/kg) before the maximum activity was confirmed. In addition, the inhibitory dose (~50%) at 1000 and 2000 μg/kg was equal to the positive control group (inhalation of P38 MAP kinase inhibitor).

This study dose analysis based on lung surface area (based on the standard used and assuming 30% intranasal dose delivered to the lungs) from 100 μg/kg, 300 μg/kg, 1000 μg/kg and 2000 μg The mouse dose of /kg provides human equalization doses of 2.43 mg, 7.3 mg, 24.3 mg, and 48.6 mg. Analysis of total cell data from our study indicates that we have identified a clear dose response relationship. The lowest dose of 100 μg/kg significantly reduced the total cell count, However, viewing the distribution in terms of individual animals clearly indicates that some animals respond very well and others are not. At 300 μg/kg, the overall mean effect was similar (about 26% and 23% reduction for 100 μg/kg) and there were relatively few changes between animals. This may suggest that the 100 μg/kg dose represents the beginning of the dose response curve and that 300 μg/kg has a clearly established effect. Similar responses were observed at 1000 μg/kg and 2000 μg/kg in quantities (50 and 46% at 1000 μg/kg and 2000 μg/kg) and data tightness, suggesting that the maximum effect was achieved at 1000 μg/kg.

This application discloses a thorough understanding of the dose-response relationship of undifferentiated heparin in this model. This provides a basic theory of choice for doses for further preclinical or clinical studies.

Example 2 (Patient samples of adult cystic fibrosis patients and frog sputum models)

The experiment was carried out using rheological and frog sputum experimental plans similar to those described by Sun et al. (2002), Rancourt et al. (2004) and O'Brien (2004). Heparin concentrations were studied and the results were compared to vehicle control (normal 0.15 M saline) and historical control (N-ethylcysteine lysinate 1 mM).

Materials and methods

The target concentration of cystic fibrosis (CF) in the in-tube sputum test is -5 mg / ml final concentration, our basic experimental plan (Sun 2002) is used to measure by magnetic rheology (King 1988) 10 The viscoelasticity of the -15 mg aliquot of the sputum sample was cultured in a 10% vol/wt bismuth solution or carrier (37 ° C / 30 minutes) at the final desired concentration and again measured Viscoelasticity. The decrease in viscoelastic modulus (in logarithm) is considered to be a specific concentration of bismuth The value of the viscoelastic effect of the agent. As a result of the small dilution associated with vehicle culture, the results were collected for the vehicle control group. 0.15M (0.9%) NaCl was used as a carrier, and N-acetinyl cysteine lysinate 1 mM (N-acetamidocysteine L-lysamine, 309 μg/mL ) as a positive control group.

For experiments involving the direct application of heparin to frog mites, a modified Ringer's solution consisting of 2/3 standard non-lactic acid Ringer's solution (Baxter) and 1/3 deionized water was used as a carrier. The modified Ringer's solution is called "Frog Ringer's" liquid.

Sputum samples from adult patients with cystic fibrosis

A sample of sputum collected from patients with cystic fibrosis attending the Alberta University Medical Clinic. A suitable amount of small amount (75-100 mg per sample) is available in our refrigerator for initial testing. The patient met the general condition of mild to moderate CF lung disease, the normal sputum manufacturer, did not take Pulmozyme or other phlegm, and was chronically infected with Pseudomonas aeruginosa.痰 samples were obtained using an experimental program approved by the Health Research Ethics Board of the University of Alberta.

Rheological properties by magnetic microrheology (King 1988)

Magnetic micro-rheometers are used to measure the bulk viscosity and elasticity of microliters of mucus. A 100 μm steel ball was carefully placed in a 1-10 μL mucus sample and vibrated by an electromagnetic field gradient. The movement of this ball is tracked by the photocell. The mapping of the ball displacement to the magnetic force is used to determine the viscosity and elasticity of the mucus as a function of the applied frequency (1-100 rad/s). These rheological properties can be used to predict mucus clearance by ciliary action and by airflow interaction (King 1987). This instrument is especially suitable for several rafts because it requires a minimum of sample. Proposed study of treatment.

Study on mucociliary clearance of frog

The frog's epithelium is covered with cilia and secretions, and it removes mucus as much as the lactating trachea. Mucosal cilia clearance is sustained at a steady rate for a few hours after sacrifice and healing (King 1998). During this time, the rate of clearance of the mucosal cilia can be adjusted by agents that alter the activity of the cilia or alter the properties of the surface fluid layer (mucus and topside ciliary fluid). By waiting longer (in the bullfrog family for 1-2 days), mucus secretion ceases and ciliary activity lasts for at least 5-6 days (Rubin 2002). During this extended period, endogenous or exogenous (eg, cystic fibrosis) mucus or mucus mimics are delivered at a rate that reflects their viscoelastic properties (King 1998). Studies in the initial and extended periods can be individually described as using "non-depleted" and "depleted" frogs, referring to the presence or absence of a mucus layer covering the cilia.

Mucosal cilia rate (MCV) is measured by observing the rate of endogenous mucus movement using a calibrated magnifying glass and a horse watch. The MCV is calculated by dividing the travel distance of the marker particles by the elapsed time (mm/s). The average of five consecutive runs of each test solution was used to calculate each MCV value.

Rheology

For each of the 10 CF 痰 samples, 6 aliquots of approximately 10-15 mg each were each at 37 ° C with 0.9% NaCl or at each of the 4 concentrations of undifferentiated heparin as outlined above or The mixture was incubated with N-acetylcysteine L-lysamine (309 μg/mL) for 30 minutes. The culture solution gently forms a layer on the crucible to avoid mechanical degradation due to agitation. In order to minimize the dilution, 1 part of the mash is 10 times the final desired concentration. (vol/wt). Before and after the culture, the viscoelasticity at 10 rad/s was determined by magnetic microrheology. The hydrazine effect of each solution is defined by the reduction in the average fraction of G* (the vector sum of viscosity and elasticity) of 10 test samples (Sun 2002).

Depleted frog mites (heparin dose response)

The effect of phlegm treatment on mucociliary clearance was tested using a depleted frog model. The rate of transport of heparin-treated sputum samples on frog ticks was measured and compared to the delivery rate of control samples treated with saline vehicle or N-ethylcysteine lysinate (Sun 2002).

Rheological (heparin molecular weight effect)

Except for the N-acetylcysteine lysine control group, the basic design was similar to the above, and the system did not need to be repeated. The diheparin fractions having different molecular weights were obtained from Neopalin, Inc (Alameda, CA, U.S.A.); these were heparin decasaccharides (molecular weight 3000 Da) and heparin polysaccharide IV (molecular weight 12,000 Da).

Depletion of frog mites (heparin molecular weight effect)

For the above CF(R) samples, the rate of heparin-treated sputum samples on frogs was measured and compared to untreated control sputum (Sun 2002).

Undepleted frog

This model of endogenous mucus represents an open system in which material exchange in the epithelium changes sputum activity. This experimental plan was designed to demonstrate the mucus-actuating effect of heparin at a concentration of two concentrations of heparin that is recommended for biological activity of CF痰 in a viscoelastic and transportability test with the same concentration of lower molecular weight. Heparin fraction (ie, Neopalin ten sugar) compared to. The dose of this diheparin preparation is 16 mg/ml and 50 mg/ml, which corresponds to a nominal final mucus concentration of 1.6 and 5 mg/ml. To control the possible changes in ciliary activity during the experiment, the order of application of undifferentiated heparin and Neopalin was changed. Each experiment was initiated with a frog Ringer's solution control group and a second Ringer's solution was applied between the two groups of heparin treatments. After completion of the heparin treatment, the final positive control group, i.e., a 1 mM solution of N-acetylcysteine lysine lysinate (NAL), was applied, resulting in a nominal mucus concentration of 100 μM. The NAL treatment is placed in the final active treatment to avoid any possible interference in the comparison between the diheparin preparations.

The solution application procedure was as follows: A 5 μL volume of sputum was gently delivered to the end of the nasal cavity of the sputum (upstream with mucociliary flow) for about 2 minutes for fluid dispersion and stabilization for clearance. The removal of the charcoal-labeled particles at a 9 mm fixed displacement was then observed under a calibrated magnifying glass, from which the rate was calculated as displacement/time (mm/min). Five readings of each solution were taken during about 10-15 minutes to calculate the average rate. For any particular condition, the rate of Ringer treatment was taken as 100% and the rate of treatment per phlegm was expressed as a percentage of the previous Ringer control group.

The final concentration of heparin in the fluid of the frog's epithelial surface is determined by a number of hypotheses. The volume of mucus that fills the surface of the palate changes with resting metabolic status and time after excision. This volume was evaluated on a 50 μL scale based on a study by Festa et al., New Reconstruction of Large Bullfrogs (1997). Therefore, by adding 5 μL of mash, it is assumed that the final concentration in the epithelial fluid is ten times lower.

Statistical processing of data

Data measurements are expressed as mean ± standard error. The overall significance of viscoelasticity and scavenging was tested using a single factor variation analysis. For the comparison between the two groups, a pair of Student T-tests was used. A significant level was set at p < 0.05.

Explanation

The main variable of interest is the reduction in mucovisile viscoelasticity, expressed as delta log G* (corrected for vehicle treatment). A statistically significant reduction in log G* for a particular heparin concentration was obtained as evidence supporting the activity of phlegm. Mucosal cilia clearance (relative to the rate of clearance of sputum treated with a sputum agent after treatment with a carrier) is the second variable of interest. Treatments that reduce the cross-linked structure without destroying the basic mucus gel structure should result in improved in vitro clearance (King 1994). Homogeneous differences were tested by variation analysis and significance between groups was tested by paired t-tests. Based on previous experiments, a significant reduction in log G* organisms was achieved with a change of approximately 0.2 log units, and a significant increase in clearance organisms was achieved with a 10% increase in MCV (King & Rubin 1994).

The continuous increase in heparinized sputum activity depending on the dose is seen in the concentration range of 0.16 mg/ml to 5.0 mg/ml (final concentration of hydrazine) (Fig. 7). At the same dose range, there was a corresponding increase in mucociliary clearance in the test tubes treated with heparin (data not shown). For comparison, for the change in logG* and the change in MCV, the same sample with N-acetyl cysteinyl lysinate (NAL) at a concentration of 309 μg/mL (in the end concentration of 100 μM in the sputum) An aliquot of the treatment system was statistically indistinguishable from 5.0 mg/ml heparin.

The cilia mucociliary rate (MCV) increased in heparin solution compared to frog Ringer's solution in a dose-dependent manner (Fig. 8): at a nominal final concentration of 1.6 mg/ml, the rate was increased to Ringer's control. The group was 116.2% ± 3.6%; at 5.0 mg / ml, the MCV was further increased to 127.9 ± 3.6%. The effect of heparin decasaccharide was similar: at 1.6 mg/ml, MCV increased to 114.7% ± 2.1% of Ringer's control group; at 5.0 mg/ml, MCV reached an average of 126.3 ± 3.1%. At the lowest heparin concentration, the MCV line was very close to 100% of the Ringer's control group. With reference to the solution system NAL 100 μM, the MCV reached an average of 130.2% ± 4.8% of the control group. This MCV value was not significantly different from the MCV of 5.0 mg/ml undifferentiated heparin or heparin decasaccharide. The second Ringer's rate was 98.9% ± 1.0% of the first, with no significant difference indicating the stability of the control during the experiment.

Undifferentiated heparin and Neopalin decasaccharide stimulated the clearance of the animal mucociliary model in a dose-dependent manner. The difference in mucus-activated activity between undifferentiated heparin and heparin-decasaccharin fractions was not statistically significant. Lack of any significant effect of heparin molecular weight on mucus motility in the study range suggests that heparin is rarely diffused from the mucus layer during the observation period (approximately 15 minutes). At the highest dose, the mucus-actuating effect of undifferentiated heparin and heparin decasaccharide was similar to 100 μM of the positive reference compound N-ethylcysteine lysine lysinate and the system was indistinguishable.

Example 3

A double-blind, placebo-controlled trial of phase I/II randomization in a four-parallel group of 1:1:1:1 ratio was performed to evaluate 25 mg in hard capsules (hydroxypropyl-methylcellulose) Pre-quantitative dose (ie, 31 mg nominal dose) presented, safe, tolerated, delivered directly to the undifferentiated heparin sodium in the lungs of patients with cystic fibrosis (CF) using the Monohaler ^® Dry Powder Inhalation (DPI) device Sex, efficacy and detection efficiency.

Experimental procedure

The study period is approximately 22 months. Each patient was involved: a 4-week screening period prior to baseline (Day 1); a 4-week treatment period; and a 2-week follow-up period. The participation period for this test was approximately 10 weeks for each patient. The evaluable subjects were randomly assigned to the treatment group.

Medication formula

50 mg, 100 and 200 mg of delivered daily dose of undifferentiated heparin sodium (Bioiberica as described in Example 1) containing L-leucine (Ajinomoto) was administered by oral inhalation (individually 62 mg, The nominal daily dose of 124 mg and 248 mg is the basis). Each individual placebo capsule contains a dry powder blend of sorbate (Respitose) and L-leucine (Ajinomoto).

Study subjects (inclusion criteria)

16-year-old male or female; non-smoker; written consent received prior to any trial special procedure; CF specialist/investigator confirmed diagnosis of CF lung disease (ie, respiratory clinical symptoms and positive sweat test or disease, including mutations) The FEV1 values at baseline were estimated to be +/- +/- FEV1 values at 4 weeks prior to screening. Within 15%; due to the regular mucus production of CF; sputum cough out easily (ie, clearance) VAS score 80 mm; the amount of neutrophil elastase and/or IL-8 is higher than the detectable level and/or upper limit of the specific laboratory; appropriate contraceptive means (subject [and his/her partner] need to use Appropriate means of contraception consists of two types of contraceptives, at least one of which is a barrier method; all requirements of this experimental plan can be observed; inhalers can be used satisfactorily.

therapy group

A total of 77 randomized evaluable patients with moderate CF were included in the study. These patients are required to meet all inclusion criteria and provide written notice of consent prior to any reagent specific procedures.

Dosing and medication plan

Patients were randomized to receive one of three daily delivery doses of 50 mg, 100 mg, 200 mg of heparin inhalation powder or a matching placebo; 4 capsules were administered twice daily by the patient by inhalation (ie, total daily dose) 8 capsules) for 4 weeks. The treatment is double-blind. The medication time was 9:00 (±2 hours) (physical treatment after morning and any bronchodilator treatment) and 21 pm (±2 hours). Note: Patients are advised on the morning of each visit (ie, baseline [Day 1], Week 2, and Week 4), and his/her morning pharmacy is evaluated after all treatments are completed. Take it. In the four groups, one group received 8 placebo capsules (4 in the morning and 4 in the afternoon) and no active capsules, one group received 2 active capsules (1 in the morning, One was taken in the afternoon) and six placebo capsules (3 in the morning and 3 in the afternoon), and the other group received 4 active capsules (2 in the morning and 2 in the morning) It is taken in the afternoon) and 4 placebo capsules (2 in the morning and 2 in the afternoon), and the last group receives 8 active capsules (4 in the morning and 4 in the system) Take it in the afternoon).

Includes five visits: 1. Screening (Day 28 [± 4 days]); 2. Baseline (1st) Day); 3. Week 2 (Day 15 [± 2 days]); 4. Week 4 (Day 29 [± 2 days]); 5. Week 6 follow-up (Day 43 [± 4 days] ).

Safety and tolerability parameters

Monitoring and evaluation: AE/severe adverse events (SAE); sitting vital signs parameters (blood pressure, heart rate, frequency of absorption, temperature), weight and physical examination; clinical laboratory parameters, including hematology, clinical chemistry, and urine Examination; platelet counts were measured at screening, baseline, week 2, and week 4.

Pharmacodynamic (PD) parameters:

Measurement parameters of coughing sputum, (ie, rheological viscoelastic / physicochemical measurement parameters); induced sputum labeled neutrophil elastase, interleukin (IL-6 and IL-8) and cytometry (ie, total cytometry) , neutrophil hematocrit %, macrophage %); EBC pH can be measured in several selected locations in related experiments; plasma labeling, neutrophil elastase, neutrophil elastase/AAT complex, Neutrophil white blood cell count, IL-6, IL-8 and CRP; aPTT and platelet count.

Main test objectives:

To assess the safety and tolerability of heparin inhalation powder in patients with CF.

Secondary test purposes:

Studying the pharmacodynamics (PD) of heparin inhaled powder by coughing mucus properties (ie, rheological properties, including viscoelastic/physicochemical measurements); labeling of inflammatory markers that induce sputum (ie, neutrophil elastase, interleukins) [IL-6 and IL-8]) and cell count (ie, total cell count, neutrophil count, % macrophage); efficacy on the pH of expiratory concentration (EBC) Selective center measurement); efficacy of inflammatory markers in plasma (ie, neutrophil elastase, neutrophil elastase/ α -1 antitrypsin [AAT] complex, IL-6, IL- 8. Neutrophil white blood cell count, and the amount of C-reactive protein [CRP]; and the effect on blood coagulation (ie, activated partial thrombin time [aPTT] and platelet count).

Evaluate efficacy by:

Complete visual analogue scale (VAS) based on the following symptoms: cough solution; cough removal, consistency, volume, color, viscoelasticity (viscosity); difficulty breathing; overall well-being (including feeling, energy, body) Activity, appetite, and sleep); microbiological analysis of the effects of coughing on bacterial growth, density, and antibiotic susceptibility (including identification, culture, density, and antibiotic susceptibility of the following organisms: Pseudomonas aeruginosa [mucus and other types) ], Burkberry onion, Haemophilus influenzae, and Staphylococcus aureus; volume and weight of a 24-hour cumulative cough sample; forced exhalation in 1 second (FEV1), forced vital capacity (FVC), Forced expiratory mid-flow (FEF25-75), and pulmonary function measured by arterial oxygen saturation (SaO2); and response to cystic fibrosis investigation (CFQ);

data analysis

Patients randomized to placebo were pooled together. Merged statistics Produced by the delivered dose, ie, 0 mg (placebo), 50 mg, 100 mg, and 200 mg. The frequency and percentage of treatment for emergency adverse events (TEAE), related TEAEs and SAEs encoded by the MedDRA are summarized by systematic organ classification and preferred terms. Appropriate summaries of clinical laboratory parameters, vital signs, weight, and physical examinations were presented by respondents. Baseline (Day 1) to Week 4 at VAS (cough solution, cough and sputum clearance, consistency, volume, color, viscoelasticity [viscous]; dyspnea; 24 hours cumulative coughing sample weight and volume, Overall well-being (including sensation, energy, physical activity, appetite, and sleep), inflammatory markers, mucus rheology measurements, microbiological measurements (cell growth in weaving organisms, density and resistance to susceptibility), and lung function tests ( Changes in FEV1, FVC, and FEF25-75) were analyzed using a baseline amount as a covariate covariate (ANCOVA) model. For each active dose versus placebo comparison, the difference between doses was assessed at 95%. The confidence interval is presented.

result Safety (local and systemic) and tolerance

The mean platelet count after administration of 50 mg, 100 mg, and 200 mg of undifferentiated heparin was compared to lactose placebo and demonstrated no systemic exposure after heparin administration throughout the study period (see Figure 9).

Similarly, the mean activated partial thrombin time (aPTT) after administration of 50 mg, 100 mg, 200 mg of delivered undifferentiated heparin compared with lactose placebo confirmed no systemic exposure after heparin administration throughout the study period. (See Figure 10).

Induced sputum neutrophil elastase, total cell count and IL-8 amount

These data confirm the mean reduction in neutrophil elastase 0.3 log (compared to placebo) and mean reduction in total cell count 0.3 log (compared to placebo) for key research purposes and priority acceptance criteria, see Figures 11 and 12. The 0.3 log reduction is clinically relevant and is associated with pulmonary function maintenance in CF patients.

As shown in Figure 13, IL-8 levels were reduced by administration of heparin relative to placebo and demonstrated a trend toward low inflammation.

Overview of plasma inflammatory mediators

These data confirm that no systemic effects were observed after oral administration of heparin, which was labeled with plasma (eg, plasma neutrophil count, neutrophil elastase/ α -1 antitrypsin complex) , CRP, IL-8 and IL-6 - see Figures 14 to 20) measurements.

痰 Clearance and 痰 color (VAS score)

As demonstrated by the data in Figure 21, each dose provided to the patient showed improved efficacy when compared to the placebo group. The effect of heparin on reducing inflammation and alleviating CF symptoms is shown by the effect of the patient's sputum color (less sputum color suggests less infection) and sputum clearance.

Hemoptysis

Heparin is an anti-mimetic agent, and in some groups of patients (including CF patients, which have a tendency to suffer from hemoptysis (pulmonary bleeding)) can cause bleeding.

Although previous studies using aerosolized heparin have confirmed no increase in hemoptysis, it is important to note that these nebulized solutions use heparin as a drip, which granules throughout the upper respiratory tract but does not reach the lungs (end of the lungs). . Because the end of the lung contains a large number of vascular structures and therefore contains the possibility of hemoptysis, the atomization formula is not expected Caused a lot of hemoptysis. However, based on the information currently available, if heparin does not reach the lower reaches of the lungs, it is expected to cause hemoptysis. We have surprisingly found that this is not the case.

In the present study, a significant ratio of heparin delivered to a patient can reach the bronchial end of the lung (eg, a dose of 10 mg of fine particles per 25 mg of heparin delivered dose). Based on the results of this study, 7% of patients receiving heparin reported hemoptysis compared with 5% of patients receiving placebo. As described above, based on the anticoagulant properties of heparin and the hemoptysis of CF patients, this result is surprising.

cough

Cough in low (11% of patients) and high (10% of patients) doses were reported with placebo (15% of patients), but no coughing was reported in patients taking intermediate doses. In previous clinical studies, the incidence of more than 20% of patients was reported to cough due to the formula used.

Figure 1 shows the total cells obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The number is reduced.

Figure 2 shows the total amount obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The number of phagocytes is reduced.

Figure 3 shows the total epithelial obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The number of cells is reduced.

Figure 4 shows the total sensation obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The decrease in the number of neutral white blood cells.

Figure 5 shows the total sensation obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The number of acid white blood cells is reduced.

Figure 6 shows the total lymph obtained from BAL after administration of heparin doses of 100 μg/kg, 300 μg/kg, 1000 μg/kg, and 2000 μg/kg compared with the sham-operated control group and the positive control group ADS115398. The number of cells is reduced.

Figure 7 shows the administration of 0.16 mg/ml, 0.5 mg/ml, 1.6 mg/ml and 5 mg compared with the control group, the saline solution and the positive control group N-acetylcysteine lysine lysinate. /ml of heparin, the change in viscoelasticity of mucus.

Figure 8 shows the administration of 0.5 mg/ml, 1.6 mg/ml and 5 mg/mg compared to 0.5 mg/ml, 1.6 mg/ml and 5 mg/ml (4-6 strips) of low molecular weight heparin. Changes in mucovisile viscoelasticity (relative rate (% of Ringer's control)) in milliliters (lanes 1-3) of undifferentiated heparin, and n-acetyl cysteinyl lysine The acid salt (7th strip) demonstrates an increase in the dose-dependent increase in mucociliary rate and compatibility with clinically relevant N-acetylcysteine lysine lysine agents.

Figure 9 shows the mean platelet count after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data confirmed that no systemic exposure was present after administration of heparin throughout the study period.

Figure 10 shows the mean active partial thrombin time (aPTT) after administration of 50 mg, 100 mg, 200 mg of undifferentiated heparin compared to lactose placebo. The data confirmed that no systemic exposure was present after administration of heparin throughout the study period.

Figure 11 shows an overview of induced neutrophil elastase after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data confirmed that the amount of neutrophil elastase decreased after the 100 mg dose.

Figure 12 shows an overview of induced total cell counts after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data demonstrates a decrease in total cell count after administration of undifferentiated heparin compared to lactose placebo.

Figure 13 shows an overview of induced 痰IL-8 after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data demonstrates a reduction in IL-8 levels after administration of a 100 mg dose of heparin compared to lactose placebo.

Figure 14 shows a summary of plasma neutrophil elastase following administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data confirmed no plasma inflammatory markers.

Figure 15 shows an overview of plasma neutrophil counts after administration of 50 mg, 100 mg, 200 mg of delivered undifferentiated heparin compared to lactose placebo. The data confirmed no plasma inflammatory markers.

Figure 16 shows plasma NE/AAT mismatch after delivery of 50 mg, 100 mg, 200 mg of undifferentiated heparin compared to lactose placebo. Overview of things. The data confirmed no plasma inflammatory markers.

Figure 17 shows a summary of plasma CRP after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data confirmed no plasma inflammatory markers.

Figure 18 shows a summary of plasma IL-8 after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo. The data confirmed no plasma inflammatory markers.

Figure 19 shows a summary of plasma IL-6 after delivery of 50 mg, 100 mg, 200 mg of undifferentiated heparin compared to lactose placebo. The data confirmed no plasma inflammatory markers.

Figure 20 shows a summary of plasma inflammatory mediators after administration of 50 mg, 100 mg, 200 mg delivered doses of undifferentiated heparin compared to lactose placebo. The data confirms that there is no plasma inflammatory marker amount for all neutrophils, neutrophil elastase neutrophil elastase/ α -1 antitrypsin complex, IL-8, IL-6 and CRP. In Fig. 20, the first strip in the histogram indicates a low dose, the second strip indicates a medium dose, and the third strip indicates a high dose.

Figure 21 shows mucus observations after administration of 50 mg, 100 mg, 200 mg of delivered dose of undifferentiated heparin compared to lactose placebo, from baseline to week 4 of the study. The data confirmed that the patient's mucus became thinner after heparin treatment. The volume of sputum increased for the placebo system. The mucus color becomes clearer, indicating a reduction in bacterial infection.痰 becomes less viscous and therefore easier to remove from the lungs.

Figure 22 shows the change from the baseline to the 4th week of the study, with lactose The amount of S. aureus and Pseudomonas aeruginosa in the sputum after administration of 50 mg, 100 mg, and 200 mg of undifferentiated heparin compared to placebo. The data confirmed no microbial action.

In each of the histograms of Figures 21 and 22, if present, the first strip represents a low dose, the second strip represents a medium dose, the third strip represents a high dose, and the fourth strip represents a placebo.

Claims (63)

A glycosaminoglycan (eg, heparin) for treating and/or preventing an inflammatory lung disease, optionally wherein the glycosaminoglycan (eg, heparin) is decreased after administration of a drug to a subject The subject's lungs are inflamed. A method of treating and/or preventing an inflammatory lung disease comprising administering a glycosaminoglycan (eg, heparin) to a subject, optionally, wherein, after administration, the glycosaminoglycan (eg, , heparin) reduces inflammation of the lungs of the subject. A use of a glycosaminoglycan (eg, heparin) for the manufacture of a medicament for the treatment and/or prevention of an inflammatory lung disease, optionally wherein the glycosaminoglycan (eg, after administration to a subject) , heparin) reduces inflammation of the lungs of the subject. A kit comprising glycosaminoglycan for treating and/or preventing an inflammatory lung disease, optionally wherein the glycosaminoglycan reduces the subject after administration to a subject Inflammation of the lungs. The glycosaminoglycan, method, use or kit of claim 1 to 4, respectively, wherein the glycosaminoglycan is in the form of a dry powder. A glycosaminoglycan, method, use or kit according to any one of claims 1 to 5 wherein the administration of the glycosaminoglycan (e.g., heparin) causes at least a 10% reduction in inflammation. A glycosaminoglycan, method, use or kit according to claim 6 wherein the administration of the glycosaminoglycan (e.g., heparin) causes at least a 20% reduction in inflammation. Such as the glycosaminoglycan, method, use or kit of claim 7 Among them, administration of the glycosaminoglycan (for example, heparin) causes at least 30% reduction in inflammation. A glycosaminoglycan, method, use or kit according to claim 8 wherein the administration of the glycosaminoglycan (e.g., heparin) causes at least 50% reduction in inflammation. A glycosaminoglycan, method, use or kit according to claim 9 wherein the administration of the glycosaminoglycan (e.g., heparin) causes at least a 60% reduction in inflammation. The glycosaminoglycan, method, use or kit of any one of claims 1 to 10, wherein the glycosaminoglycan (e.g., heparin) is administered via the nasal cavity or by pulmonary inhalation. The glycosaminoglycan, method, use or kit of any one of claims 1 to 11, wherein the glycosaminoglycan is heparin. A glycosaminoglycan, method, use or kit according to claim 11 wherein the heparin is unfractionated heparin. The glycosaminoglycan, method, use or kit of claim 13, wherein the heparin is undifferentiated heparin sodium. The glycosaminoglycan, method, use and kit according to any one of claims 1 to 13, wherein the heparin is a low molecular weight heparin. The glycosaminoglycan, method, use or kit of any one of claims 1 to 15, wherein the total daily dose of heparin (measured as FPD) is between 0.1 mg and 6000 mg. The glycosaminoglycan, method, use or kit of claim 16, wherein the total daily dose of the heparin (measured by FPD) is 0.5 Between mg and 6000 mg. The glycosaminoglycan, method, use or kit of claim 17, wherein the total daily dose of the heparin is between 2 mg and 240 mg. The glycosaminoglycan, method, use or kit of claim 18, wherein the total daily dose of the heparin is between 5 mg and 80 mg. The glycosaminoglycan, method, use or kit of claim 19, wherein the total daily dose of the heparin is between 10 mg and 40 mg. The glycosaminoglycan, method, use or kit of claim 19, wherein the total daily dose of the heparin is between 12 mg and 50 mg. The glycosaminoglycan, method, use or kit of any one of claims 1 to 21, wherein the total daily delivery dose of the heparin is between 0.8 and 200 mg. The glycosaminoglycan, method, use or kit of any one of claims 1 to 22, wherein the heparin is delivered in a fine particle dose between 0.3 mg and 2000 mg. A glycosaminoglycan, method, use or kit according to claim 23, wherein the heparin is delivered in a fine particle dose between 0.5 mg and 2000 mg. The glycosaminoglycan, method, use or kit of claim 24, wherein the heparin is between 2 mg and 240 mg of fine particles Dosage delivery. A glycosaminoglycan, method, use or kit according to claim 25, wherein the heparin is delivered in a fine particle dose between 6 mg and 35 mg. A glycosaminoglycan, method, use or kit according to claim 23, wherein the heparin is delivered in a fine particle dose between 0.5 mg and 20 mg. The glycosaminoglycan, method, use or kit of any one of claims 1 to 27, wherein the inflammation is measured by total cell count obtained from bronchoalveolar perfusion. The glycosaminoglycan, method, use or kit of any one of claims 1 to 28, wherein the inflammation is measured by the amount of neutrophil in the lung. The glycosaminoglycan, method, use or kit of any one of claims 1 to 29, wherein the reduction is achieved by a P38 MAP kinase inhibitor, ADS115398. A glycosaminoglycan, method, use or kit according to claim 30, wherein the ADS115398 is administered to a subject by pulmonary inhalation or nasal administration at a dose of 0.1 mg/kg. The glycosaminoglycan, method, use or kit of any one of claims 1 to 31, further comprising an additional active agent. The glycosaminoglycan, method, use or kit of claim 32, wherein the additional active agent is selected from the group consisting of a bismuth agent; a bronchodilator; a scorpion toxin receptor blocker; an antibiotic and an antibacterial agent Anti-infective agent; amine Glycosides; anti-leather drugs; various anti-infective agents; non-steroidal anti-inflammatory agents; other anti-inflammatory agents; PDE4 inhibitors; quinazolinedione; steroids; matrix metalloproteinase inhibitors; (ENaC) inhibitor; methylxanthine; and drugs for cystic fibrosis management. The glycosaminoglycan, method, use or kit of claim 33, wherein the additional active agent is selected from the group consisting of a bismuth agent; a bronchodilator; a scorpion toxin receptor blocker; an antibiotic and an antibacterial agent Anti-infective agent; aminoglycoside; anti-leech drug; various anti-infective agents; non-steroidal anti-inflammatory agents; other anti-inflammatory agents; PDE4 inhibitors; quinazolinedione; steroids; Cystic fibrosis management drugs. A glycosaminoglycan, method, use or kit according to claim 34, wherein the additional active agent is ADS115398. The glycosaminoglycan, method, use or kit of any one of claims 1 to 35, wherein the inflammatory lung disease is characterized by a disease of neutrophils. The glycosaminoglycan, method, use or kit of any one of claims 1 to 36, wherein the inflammatory lung disease is selected from the group consisting of asthma, cystic fibrosis, and idiopathic pulmonary fibrosis Non-cystic fibrotic bronchiectasis, and one of the diseases or conditions of chronic obstructive pulmonary disease. The glycosaminoglycan, method, use or kit of claim 37, wherein the inflammatory lung disease is chronic obstructive pulmonary disease (COPD). The glycosaminoglycan, method, use or kit of claim 38, wherein the COPD is an acute COPD episode. The glycosaminoglycan, method, use or kit of claim 37, wherein the inflammatory lung disease is cystic fibrosis. The glycosaminoglycan, method, use or kit of any one of claims 1 to 40, wherein the glycosaminoglycan is further comprised between 1 and 20% by weight, preferably 2 to 15% by weight. One of the additives (for example, leucine). A passive or active inhaler device in combination with a glycosaminoglycan. The device of claim 42, wherein the device is a passive inhaler device. The device of claim 43, wherein the device is selected from the group consisting of Monohale and a device manufactured by Vectura (as covered by International Patent Application No. WO 2010/086285). The device of any one of claims 42 to 44, wherein the glycosaminoglycan is in the form of a dry powder. The device of any one of claims 42 to 44, wherein the glycosaminoglycan is heparin. A device for administering a glycosaminoglycan (eg, heparin) for treating and/or preventing an inflammatory lung disease, optionally wherein the glycosaminoglycan is reduced after administration to a subject The lungs of the lungs of the subject. The device of claim 47, wherein the device is a passive inhaler device. The device of claim 48, wherein the device is selected from the group consisting of Monohale and devices manufactured by Vectura (as covered by International Patent Application No. WO 2010/086285). The device of any one of claims 47 to 49, wherein the glycosaminoglycan is in the form of a dry powder. The device of any one of claims 47 to 50, wherein the glycosaminoglycan is heparin. Use of a drug delivery device for delivering a glycosaminoglycan pharmaceutical formulation of a glycosaminoglycan (eg, heparin) for treating and/or preventing an inflammatory lung disease, optionally, wherein Upon glycosaminoglycan administration, the glycosaminoglycan reduces inflammation in the lungs of the subject. The device of claim 52, wherein the device is a passive inhaler device. The device of claim 53, wherein the device is selected from the group consisting of Monohale and a device manufactured by Vectura (as covered by International Patent Application No. WO 2010/086285). The device of any one of claims 52 to 54, wherein the glycosaminoglycan is in the form of a dry powder. The device of any one of claims 52 to 55, wherein the glycosaminoglycan is heparin. A pharmaceutical formulation of glycosaminoglycans. A pharmaceutical formulation of a glycosaminoglycan (eg, heparin) for treating and/or preventing an inflammatory lung disease, optionally wherein the glycosaminoglycan reduces the test after administration to a subject The inflammation of the lungs. a pharmaceutical formulation according to claim 57 or 58 wherein the sugar amine The polysaccharide is a dry powder. The pharmaceutical formulation according to any one of claims 57 to 59, wherein the glycosaminoglycan is heparin. The pharmaceutical formulation according to any one of claims 57 to 60, wherein the formulation further comprises 1 to 20% by weight or preferably 2 to 15% by weight of one of the additives (for example, leucine). The pharmaceutical formulation of any one of claims 57 to 61, wherein the formulation further comprises a carrier. The pharmaceutical formulation of any one of claims 57 to 62, wherein the glycosaminoglycan comprises from 80 to 99% by weight of the formulation.