WO2022005321A1 - Inhaled tocilizumab for treating interleukin-6 related respiratory diseases - Google Patents

Abstract

The present invention is in the field of healthcare. More specifically, the invention relates to the use of inhaled tocilizumab and aqueous pharmaceutical compositions thereof in the treatment of interleukin-6 related respiratory diseases. The invention provides tocilizumab for use in the treatment of an interleukin-6 related respiratory disease in a mammal by pulmonary administration in an amount of 0.1 to 100.0 mg per kg body weight of the mammal, wherein the disease is selected from the group consisting of asthma, pneumonia, pulmonary edema, chronic obstructive pulmonary disease, and acute respiratory distress syndrome. Further, the invention provides a method of treating an interleukin-6 related respiratory disease, which comprises a step of pulmonary administration of tocilizumab to a mammal in need thereof in an amount of 0.1 to 100.0 mg per kg body weight of the mammal.

Description

INHALED TOCILIZUMAB FOR TREATING INTERLEUKIN-6 RELATED RESPIRATORY

DISEASES

Field of the Invention

[0001] The present invention is in the field of healthcare. More specifically, the invention relates to the use of inhaled tocilizumab in the treatment of interleukin-6 related respiratory diseases.

Background of the invention

[0002] lnterleukin-6 (IL-6) is a small size cytokine (21 KDa) produced by cells from the innate immune system and non-immune cells such as pulmonary epithelial cells. IL-6 is essential for regulation of acute phase immune process, but overproduction of IL-6 contributes to the pathogenesis of a variety of respiratory diseases. Overexpression of IL-6 in pulmonary epithelial cells was observed in patients with asthma and other respiratory diseases. Rincon M, Irvin CG. Role of IL-6 in asthma and other inflammatory pulmonary diseases. Int J Biol Sci. 2012;8(9):1281-90. Increased levels of IL-6 were found in asthmatic patients. Yokoyama A et al. Am J Respir Crit Care Med. 1995 May;151(5):1354-8. Elevated IL-6 levels were found to be predictive of increase mortality in chronic obstructive pulmonary disease (COPD) patients. Celli BR et al.. Am J Respir Crit Care Med. 2012, 185(10): 1065-72. Elevated IL-6 levels are associated with COVID-19 and other virus infection related acute respirtory distress-syndrome (ARDS) and pneumonia. McGonagle D et al. Autoimmun Rev. 2020, 19(6):102537. Liu B et al. J Autoimmun. 2020, 111:102452. IL-6 early predicts in-hospital mortality for patients wth COVID- 19. Luo M et al. JCI Insight. 2020:139024. IL-6 is a strong discriminator for severe disease in COVID-19. Henry BM et al. Clin Chem Lab Med. 2020, 58(7):1021 -1028. Elevated IL-6 levels in patients with COVID-19 are predictor of most severe course of the disease and the need for intensive care. Gubernatorova EO et al. Cytokine Growth Factor Rev. 2020, 53:13-24. IL-6 was elevated in influenza A/H1N1 virus pneumonia. Davey RT Jr et al. PLoS One. 2013;8(2):e57121. Elevated IL-6 levels in patients with community-acquired pneumonia are predictor for the severity of the disease. Ramirez P et al. Crit Care Med. 2011, 39(10) .2211-7. High IL-6 concentrations in serum and bronchoalveolar lavage fluid in pneumonia patients are associated with the disease severity and poor outcome. Bordon J et al. Int J Infect Dis. 2013, 17(2):e76-83. The concentrations of IL-6 were associated with early mortality of patients with community-acquired pneumonia. Bacci MR et al. J Med Biol Res. 2015, 48(5):427-32. The concentrations of IL-6 reflected the severity of disease in patients with community-acquired pneumonia. Zobel K et al. BMC Pulm Med. 2012, 12:6. The concentrations of IL-6 were higher in nonsurvivors than in survivor patients with community-acquired pneumonia. Lee YL et al. J Crit Care. 2010, 25(1):176.e7-13. Increased IL-6 levels were associated with pulmonary fibrosis. Papins SA et al. Cytokine. 2018, 102:168-172. Elevated IL-6 levels increases risk of death in hospitalized patients with pneumonia. Andrijevic I et al. Ann Thorac Med. 2014, 9(3):162-7. Given that respiratory diseases represent a significant source of mortality and morbidity and that overexpression of IL-6 is the pathogenic factor in such diseases, there is a great need in safe and effective approaches to antagonize IL-6 activity in patients with respiratory diseases.

[0002] Tocilizumab is a recombinant humanized anti-human interleukin 6 receptor monoclonal antibody of the immunoglobulin lgG1 subclass with a molecular weight of approximately 148 kDa. European patent 0628639B; US patent 5,795,965; Japanese patent 3370324B. Tocilizumab binds to both membrane (mlL-6R) and soluble (slL-6R) interleukin-6 receptors (IL- 6R), thereby antagonizing action of IL-6 in patients with rheumatoid arthritis, systemic juvenile idiopathic arthritis, Castleman's disease, giant cell arteritis, and cytokine release syndrome. China's National Health Commission included the use of tocilizumab in guidelines to treat coronavirus disease 2019 (COVID-19) patients. Tocilizumab has been shown to reduce the severity of COVID-19 disease and mortality in COVID-19 patients. Zhang C et al. Int J Antimicrob Agents. 2020, 55(5):105954. Xu X et al. Proc Natl Acad Sci USA. 2020, 117(20): 10970-10975. Conventionally, tocilizumab is used in form of subcutaneous injections and intravenous infusions.

[0003] Pulmonary drug delivery represents a drug administration method that provides direct and fast topical treatments for respiratory diseases. Compared to intravenous or subcutaneous injections, it provides a painless and safer alternative. Nothing is known about efficacy of inhaled tocilizumab in the treatment of IL-6 related respiratory diseases in comparison with conventional injection routes of tocilizumab administration.

[0004] Surprisingly, we found that inhaled tocilizumab is more effective in antagonizing IL-6 activity in a case of respiratory diseases compared to tocilizumab administered in the same dose by injections.

[0005] It is an object of the present invention to provide tocilizumab for use in the treatment of interleukin-6 related respiratory diseases by pulmonary administration.

[0006] It is another object of the present invention to provide an aqueous pharmaceutical composition comprising tocilizumab for use in the treatment of interleukin-6 related respiratory diseases by pulmonary administration. [0007] It is another object of the present invention to provide a method of treating interleukin-6 related respiratory diseases, which comprises a step of pulmonary administration of effective amounts of tocilizumab to a mammal in need thereof.

Summary of the invention

[0008] The present invention provides tocilizumab for use in the treatment of an interleukin-6 related respiratory disease in a mammal, in need thereof, by pulmonary administration.

[0009] A first aspect of the invention relates to tocilizumab for use in the treatment of an interleukin-6 related respiratory disease in a mammal by pulmonary administration in an amount of 0.1 to 100.0 mg per kg body weight of the mammal.

[0010] According to another aspect, the invention relates to an aqueous pharmaceutical composition comprising tocilizumab and a pharmaceutically acceptable excipient for use in the treatment of an interleukin-6 related respiratory disease in a mammal by pulmonary administration of said composition in a dose containing 0.1 to 100.0 mg tocilizumab per kg body weight of the mammal.

[0011] According to another aspect, the invention relates to a method of treating an interleukin-6 related respiratory disease, which comprises a step of pulmonary administration of tocilizumab to a mammal in need thereof in an amount of 0.1 to 100.0 mg per kg body weight of the mammal.

[0012] In preferred embodiments, the interleukin-6 related respiratory disease is selected from the group consisting of asthma, pneumonia, pulmonary edema, chronic obstructive pulmonary disease, and acute respiratory distress syndrome.

[0013] In preferred embodiments, tocilizumab or formulations or composition thereof are administered pulmonary in form of aerosol having particle size from 0.5 to 10.0 microns, more preferably from 1 to 5 microns.

[0014] Because of pulmonary route of administration, inhaled tocilizumab of the present invention have an advantage over conventional tocilizumab injections in that inhaled tocilizumab is more effective in the treatment of interleukin-6 related respiratory diseases.

[0015] Because of pulmonary route of tocilizumab administration, this invention provides particularly advantageous composition and methods for achieving the therapeutic effect in a mammal suffering from an interleukin-6 related respiratory disease. Brief Description of the Drawings

[0016] FIG.1 shows hematoxylin-eosin stained histological preparations of healthy lung tissue (0 h) and lung preparations obtained from animals after 24, 72, and 120 h of administering lipopolysaccharide (LPS), as described in example 2 of the invention.

[0017] FIG.2 shows the Kaplan-Meier survival curves for mice with acute lung injury treated with inhaled tocilizumab in comparison with tocilizumab injection at the same dose, as described in example 2 of the invention.

Detailed description of the invention

[0018] The present invention provides tocilizumab for use in the treatment of an interleukin-6 related respiratory disease in a mammal by pulmonary administration in an amount of 0.1 to 100.0 mg per kg body weight of the mammal.

[0019] As used herein, the term “tocilizumab” refers to a recombinant humanized anti-human interleukin-6 receptor (IL-6R) monoclonal antibody of the immunoglobulin lgG1 subclass that antagonizes interleukin-6 action in a mammalian body upon binding to both membrane (mlL-6R) and soluble (slL-6R) interleukin-6 receptors.

[0020] As used herein, the term “interleukin-6 (IL-6)” refers to native interleukin-6 from any species, including mouse, rat, bovine, and human, preferably human.

[0021] As used herein, the term “interleukin-6 related respiratory disease” refers to a respiratory disease associated with interleukin-6 overproduction. As used herein, the term “respiratory disease” refers to a disease affecting the respiratory system including asthma, chronic bronchitis, bronchiectasis, chronic obstructive pulmonary disease (COPD), pneumonia, COVID- 19 related pneumonia, acute respiratory distress-syndrome (ARDS), COVID-19 related ARDS, a virus related ARDS, a virus related pneumonia, pulmonary edema, and pulmonary fibrosis.

[0022] In preferred embodiments, the interleukin-6 related respiratory disease is selected from the group consisting of asthma, pneumonia, pulmonary edema, chronic obstructive pulmonary disease, and acute respiratory distress syndrome. [0023] As used herein, the term “pulmonary administration” refers to administration of tocilizumab or a formulation thereof through the lungs by inhalation. As used herein, the term “inhalation” refers to inhaling the vapor or dispersion of solid or liquid particles with added medication. In specific examples, inhaling can occur through a nebulizer or other aerosol- delivery device. The term “inhalation” used with respect to formulations and compositions of the invention is synonymous with “pulmonary administration.”

[0024] As used herein, the term “treatment” or “treating” means to reverse, alleviate, or inhibit the progress of a disease to which this term is applied, or one or more symptoms of this disease. The term “treatment” refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already with the disorder as well as those prone to having the disorder or diagnosed with the disorder or those in which the disorder is to be prevented.

[0025] As used herein, “mammal” refers to any animal classified as a mammal, e.g. mouse, rat, bovine, and human. The preferred mammal herein is a human.

[0026] In some embodiments, tocilizumab is administered by inhalation in effective amounts. As used herein, the term “effective amount” refers to an amount of an active pharmaceutical ingredient (API) or formulations or compositions thereof that is sufficient to cause a reduction, reverse, alleviation, or inhibition of progress of a disease.

[0027] In preferred embodiments, tocilizumab is pulmonary administered in an amount of 0.1 to 100.0 mg per kg body weight of a mammal.

[0028] In some embodiments, tocilizumab or formulations or compositions thereof will be administered pulmonary in form of aerosol. Numerous methods and devices are well-known from the art that can be employed to generate said aerosols in therapeutically useful size ranges and concentrations. Specifically, these are nebulizers, metered-dose inhalers (MDIs), and dry powder inhalers (DPIs). Pilcer G et al. Int J Pharm. 2010, 392(1 -2):1 -19. Nebulizers such as jet nebulizers or ultrasonic nebulizers are used for the delivery of aqueous pharmaceuticals. MDIs suspends or dissolves drug powders into liquid propellants and when a metered quantity of the propellant is released from the storage canister, the propellant evaporates and expands quickly to disperse the powdered drug or liquid droplet drug. Such propellants include, but are not limited to, chlorofluorocarbon, a hydrochlorofluorocarbon, or a hydrocarbon. DPIs delivers a precisely measured dose medicine into the lungs in dry powder form. It is designed to generate a drug powder aerosol onto or via the inspiratory air flow. In practicing the invention, any methods and devices, including nebulizers, MDIs, and DPIs may be used for pulmonary delivery of tocilizumab or pharmaceutical compositions thereof to a patient in need thereof.

[0029] As used herein, the term “aerosol” refers to suspension of liquid or solid particles in a gaseous medium. In practicing the invention, tocilizumab or compositions thereof can be administed pulmonary as aerosol in form of liquid formulations and dry powders.

[0030] In some embodiments, tocilizumab is pulmonary administered in form of aerosol with particle size of 0.5 to 10.0 microns, preferably of 0.6 to 5.0 microns.

[0031] In practicing the invention, tocilizumab will typically be administered as a liquid or dry formulation in association with one or more pharmaceutically acceptable excipients.

[0032] The present invention provides an aqueous pharmaceutical composition comprising tocilizumab and a pharmaceutically acceptable excipient for use in the treatment of an interleukin-6 related respiratory disease in a mammal by pulmonary administration of the composition in a dose containing 0.1 to 100.0 mg tocilizumab per kg body weight of the mammal.

[0033] As used herein, the term "excipient" describes any ingredient other than tocilizumab. Such excipients include, but are not limited to, pharmaceutical water such as purified water or water for injections; buffer system such as phosphate buffer to maintain pH values from 3.0 to 8.5, preferably of about 6.5; ionic, non-ionic, and amphiphilic surfactant or combinations thereof; suitable osmotically active inorganic agent such as chlorides, sulfates or phosphates of sodium, calcium or magnesium; suitable osmotically active organic agent such as sugars and sugar alcohols, in particular trehalose, mannitol, and sorbitol; preservatives and antioxidants.

[0034] In preferred embodiments, the excipient is selected from the group consisting of polysorbate 80, sucrose, disodium phosphate dodecahydrate, sodium dihydrogen phosphate dehydrate, L-histidine, L-histidine monohydrochloride, L-arginine, L-arginine hydrochloride, and L-methionine.

[0035] As used herein, the term “aqueous pharmaceutical composition” means that a pharmaceutical water, e.g. purified water or water for injections, is the essential excipient of the composition of the invention. [0036] In practicing the invention, the aqueous pharmaceutical composition may be in a unit dosage form suitable for a single pulmonary administration of a precise dose. As used herein, the term "unit dosage form" refers to a physically discrete unit of the composition of the invention suitable for a mammal to be treated by pulmonary route of administration. The level of a particular effective dose for any particular mammal depends on many factors, including the kind of the mammal; disorder being treated and the severity of the disorder; the particular composition used; age, weight, general health, gender and diet of the subject; time of administration; duration of treatment; drugs and/or additional therapies combined with or in combination with the composition of the present invention, and similar factors well known in medical technology.

[0037] In some embodiments, the unit dosage form contains tocilizumab in amount of 1.0 to 400.0 mg.

[0038] In practicing the present invention, the composition of the invention can be prepared by procedures well-known from the art. Such procedures include, but are not limited to, mixing tocilizumab with other ingredients of the composition in conventional manner. Guidance for the preparation of compositions of the invention can be found in "Remington: The science and practice of pharmacy" 20th ed. Mack Publishing, Easton PA, 2000 ISBN 0-912734-04-3 and " Encyclopaedia of Pharmaceutical Technology", edited by Swarbrick, J. & J. C. Boylan, Marcel Dekker, Inc., New York, 1988 ISBN 0-8247-2800-9 or a newer edition.

[0039] The present invention provides a method of treating an interleukin-6 related respiratory disease, which comprises a step of pulmonary administration of tocilizumab to a mammal in need thereof in an amount of 0.1 to 100.0 mg per kg body weight of the mammal.

[0040] In some embodiments, tocilizumab or pharmaceutical compositions thereof can be administered pulmonary for one day or longer. Preferably, tocilizumab or pharmaceutical compositions thereof can be administered pulmonary once per four weeks.

[0041] In practicing the method of the invention, the amount of tocilizumab to be administered pulmonary depends on the kind of mammals. Equivalent doses for humans can be calculated from doses for other mammals, and vice versa, as described in, e.g. Guidance for Industry Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers. U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER), 2005, Pharmacology and Toxicology. The guidance teaches that a human equivalent dose can be obtained by dividing an animal dose by certain factor, e.g. by 12.3, 6.2, and 1.1 for mouse, rat, and mini-pig, respectively.

[0042] In practicing the method of the invention, tocilizumab may be administered as an active pharmaceutical ingredient of an aqueous pharmaceutical composition comprising tocilizumab and a pharmaceutically acceptable excipient.

[0043] Because of pulmonary route of administration, inhaled tocilizumab of the present invention have an advantage over conventional tocilizumab injections in that inhaled tocilizumab is more effective in the treatment of interleukin-6 related respiratory diseases.

[0044] Because of pulmonary route of tocilizumab administration, this invention provides particularly advantageous composition and methods for achieving the therapeutic effect in a mammal suffering from an interleukin-6 related respiratory disease.

[0045] The following examples are presented to demonstrate the invention. The examples are illustrative only and are not intended to limit the scope of the invention in any way.

I

Example 1

[0046] This example shows the aqueous pharmaceutical composition for pulmonary administration.

[0047] Table 1

[0048] The ingredients are mixed in the conventional manner in amounts as indicated in Table 1 to prepare a sterile, preservative-free solution for further dilution prior to pulmonary administration. Single-dose vials for pulmonary administration contains 80 mg/4 ml_, 200 mg/10 mL, or 400 mg/20 mL of tocilizumab. The aqueous pharmaceutical composition is administered in recommended dose to a mammal in need thereof by inhalation using nebulizer for 30 to 90 min, once a period of four weeks. Example 2

[0049] This example illustrates advantages of inhaled tocilizumab over conventional tocilizumab injections in the treatment of interleukin-6 related respiratory diseases.

[0050] Comparative efficiencies of same doses of tocilizumab administered by pulmonary route and tocilizumab administered by conventional injections were tested in a model of acute lung injury and acute respiratory distress-syndrome (ARDS) induced by intratracheal administration of lipopolysaccharide (LPS) as described in D'Alessio FR. Mouse Models of Acute Lung Injury and ARDS. Methods Mol Biol. 2018, 1809:341-350 with some modifications aimed to make the model more lethal. For this purpose, C57BI/6J male mice were pre-treated with 1 pg/mouse of a-Galactosyl Ceramide (KRN7000, Sigma-Aldrich), as described in Aoyagi T et al. Int Immunol. 2011, 23(2):97-108, and after 24 hours mice received intratracheal LPS (E.coli; 300 pg/mouse) with addition of 10 pL/mouse Freund's complete adjuvant and 100 pg/mouse muramyl peptide. This model is characterized by dramatic overproduction of interleukin-6 in lung tissue and acute rise of lung IL-6 levels up to 5150 pg/mL that is by 43-times higher than IL-6 levels in the lungs of healthy mice (119 pg/ml). The IL-6 overproduction in this model is associated with acute lung injury, pulmonary edema, acute respiratory disitress-syndrome, and pneumonia. The IL-6 related respratory disease was accompanied with inhibition of motor activity, impairment of coordination, insuffitiently wide opening of the eyelids and ptosis, low muscle tonus, decrease in appetite, water intake, and urination; and disturbance of the respiration. FIG.1 shows hematoxylin-eosin stained histological preparations of healthy lung tissue before LPS administration (0 h) and lung preparations obtained from animals after 24, 72, and 120 h of administering lipopolysaccharide (LPS). The healthy lung preparations are characterized by moderate blood supply, the absence of pathological changes in the stroma and parenchyma, as well as in the walls of the bronchi and bronchioles. The preparations obtained from lungs after intraracheal administration of LPS show characteristic signs of acute lung injury caused by inflammation. Thus, this model reproduces the essential signs of interleukin-6 related respiratory diseases in humans such as acute respiratory distress syndrome, pulmonary edema, and pneumonia.

[0051] Mice with LPS-induced lung injury were randomized into three groups of 20 animals each and received treatments 30 min after the LPS injection. In the first group, mice received once saline s.c. (control). In the second group, mice received once tocilizumab subcutaneously at dose of 28 mg/kg (“LPS+T sc”) in form of aqueous composition of example 1. In the third group, mice received once tocilizumab by inhalation at dose of 28 mg/kg (“LPS+T inh”) in form of aqueous composition of example 1. In one else group ("Intact”), C57BI/6 mice (n=20) received no treatment. The LPS-induced acute lung injury resulted in death of most animals in control group. FIG.2 shows the Kaplan-Meyer survival curves for 456 hours in four groups. There was a statistically significant difference between the groups (Mantel-Cox logrank test; p<0.0001). The median survival, i.e. the time during which half of the animals were still alive, was 72, 48, and 72 hours in control, “LPS+T sc”, and “LPS+T inh”, respectively. After 456 hours, 15% (3/20), 30% (6/20), and 40% (8/20) animals survived in control group, “LPS+T sc” group, and “LPS+T inh” group, respectively. For reference, all animals in “Inact” group were still alive at 456 h. Comparison of survival curves for “LPS+T sc” and “LPS+T inh” groups provides the hazard ratio (Mantel-Haenszel) of 1.55 (95% Cl 0.63 to 3.79), i.e. the rate of deaths in “LPS+T inh” group was about 1.5 times less compared to the rate in “LPS+T sc” group. Thus, inhaled tocilizumab have an advantage over conventional tocilizumab injection in that the inhaled tocilizumab was more effective in the improvement of survival of mammals with IL-6 related acute lung injury compared to tocilizumab injection in the same dose.

Example 3

[0052] This example illustrates advantages of inhaled tocilizumab over conventional tocilizumab injections in reducing excessive IL-6 levels in lungs of mammals suffering from interleukin-6 related respiratory disease.

[0053] Mice with LPS-induced acute lung injury as described in example 2 were randomized into three groups of 10 animals each and received treatments 30 min after the LPS injection. In the first group, mice received once saline by subcutaneous injection (control). In the second group, mice received once tocilizumab subcutaneously at dose of 28 mg/kg (“LPS+T sc”) in form of aqueous composition of example 1. In the third group, mice received once tocilizumab by inhalation at dose of 28 mg/kg (“LPS+T inh”) in form of aqueous composition of example 1. In one else group (“Intact”), C57BI/6 mice (n=10) received no treatment. Changes of concentrations of IL-6, interleukin-1 beta (IL-b), interleukin-17A (IL-17A), tumor necrosis factor alpha (TNFa), and interferon gamma (IFNg) in lung homogenates within 12 days were monitored with the Bio-Plex Pro™ Mouse Cytokine Th17 Panel A 6-Plex immunoassay using Bio-Plex® MAGPIX™ Multiplex Reader (Bio-Rad, SN: 12250707). The results are presented in tables 2 through 6 as mean ± SEM of concentrations (pg/ml).

[0054] Table 2.

‘Differs significantly of control (p<0.05).^#Differs significantly of “LPS+T sc” (p<0.05).

[0055] Table 2 shows that LPS induced an acute rise of IL-6 levels at 2 h with subsequent gradual decrease in IL-6 levels at 12 day in mice that were still alive (control group). For reference, levels of IL-6 in lungs of non-treated healthy mice were 119 pg/mL. Tocilizumab injection resulted in a significant acute increase in lung IL-6 levels in LPS-induced mice compared even to control (p<0.05). IL-6 levels in lungs at 2 hours after the tocilizumab injection were 3-fold higher than in contol group that is a serious adverse effect of tocilizumab injection. In the second phase, from 12 hours to 12 days, lung IL-6 levels in tocilizumab-treated mice were significantly lower than in mice of control group (p<0.05). In contrast, the pulmonary administration of tocilizumab by inhalation in the same dose did not result in the undesirable acute rise of IL-6 in lungs compared to control (p>0.05). The lung levels of IL-6 at 2 hours were 3 times less in mice who received inhaled tocilizumab (p<0.05) compared to mice treated with tocilizumab injection. This provide a serious advantage of pulmonary route of tocilizumab administration as more safe over tocilizumab injection in the treatment of a respiratory disease. In the second phase, from 12 hours to 12 days, inhaled tocilizumab decreased IL-6 in lungs to levels significantly less than those observed after tocilizumab injection (p<0.05). It means that inhaled tocilizumab more effectively reduces the levels of IL-6 in the lungs of mammals suffering from the IL-6 related respiratory disease than tocilizumab injection in the same dose. Thus, pulmonary administration of tocilizumab provides more effective and safe treatment of IL-6 related respiratory diseases than tocilizumab injections.

[0056] Table 3.

‘Differs significantly of control (p<0.05).^#Differs significantly of “LPS+T sc” (p<0.05).

[0057] Table 3 shows that LPS-induced respiratory disease is accompanied by an increase in the levels of IL-1 b in the lungs (control group). Tocilizumab injection resulted in signficant transient increase of lung IL-1 b at 2 hours and delayed increase in IL-1 b at 12 day compared to control (p<0.05). The acute and delayed increase in lung IL-1 b levels represent an adverse effect of tociliumab injection. In contrast, the pulmonary administration of tocilizumab did not cause both acute and delayed significant increase in IL-1 b compared to control (p>0.05). There was a statistically significant difference in IL-1 b levels in lung between groups of inhaled tocilizumab and tocilizumab injection at 2 hours and 12 day (p<0.05). This provides a serious advantage of pulmonary route of tocilizumab administration as more safe over tocilizumab injection in the treatment of respiratory diseases.

[0058] Table 4.

*Differs significantly of control (p<0.05).^#Differs significantly of “LPS+T sc” (p<0.05).

[0059] Table 4 shows that tocilizumab injection resulted in signficant transient increase of lung IL-17A at 2 hours and dramatic delayed increase in IL-17A at 12 day compared to control (p<0.05). The acute and delayed increase in lung IL-17A levels represent an adverse effect of tociliumab injection. In contrast, the pulmonary administration of tocilizumab did not cause both acute and delayed significant increase in IL-17A compared to control (p>0.05). There was a statistically significant difference in IL-17A levels in lung between groups of inhaled tocilizumab and tocilizumab injection at 2 hours and 12 day (p<0.05). This provides a serious advantage of pulmonary route of tocilizumab administration as more safe over tocilizumab injection in the treatment of respiratory diseases.

[0060] Table 5.

^•Differs significantly of control (p<0.05).^#Differs significantly of “LPS+T sc” (p<0.05).

[0061] Table 5 shows that that tocilizumab injection resulted in signficant transient increase of lung TNFa at 2 hours and delayed increase in TNFa at 12 day compared to control (p<0.05). The acute and delayed increase in lung TNFa levels represent an adverse effect of tociliumab injection. In contrast, the pulmonary administration of tocilizumab did not cause both acute and delayed significant increase in TNFa compared to control (p>0.05). There was a statistically significant difference in TNFa levels in lung between groups of inhaled tocilizumab and tocilizumab injection at 2 hours and 12 day (p<0.05). This provides a serious advantage of pulmonary route of tocilizumab administration as more safe over tocilizumab injection in the treatment of respiratory diseases.

[0062] Table 6.

^•Differs significantly of control (p<0.05).^#Differs significantly of “LPS+T sc” (p<0.05).

[0063] Table 6 shows that pulmonary administration of tocilizumab as well as tocilizumab injection decreased significantly lung IFNg levels at 12 day compared to control (p<0.05).

[0064] As a conclusion, pulmonary administration of tocilizumab provides more effective treatment of IL-6 related respiratory disease compared to tocilizumab injection in the same dose with respect to reducing lung IL-6 levels. Moreover, pulmonary administration of tocilizumab is more safe than tocilizumab injection, since inhaled tocilizumab does not induced the acute or delayed increase in the production of pro-inflammatory cytokines such as IL-1 b, IL-17A, and TNFa in the lung.

Example 4

[0065] This example illustrates the dose-response of inhaled tocilizumab in the treatment interleukin-6 related respiratory disease.

[0066] Mice with LPS-induced acute lung injury as described in example 2 were randomized into six groups of 6 animals each and received treatments 30 min after the LPS injection. In the first group, mice received once saline s.c. (control). In other groups, mice received once tocilizumab by pulmonary administration in form of aqueous composition of example 1 in doses 0.01, 0.1, 1.0, 10.0, and 100.0 mg/kg. After 12 hours, concentrations of IL-6 in lung homogenates were measured with the Bio-Plex Pro™ Mouse Cytokine Th17 Panel A 6-Plex immunoassay using Bio-Plex® MAGPIX™ Multiplex Reader (Bio-Rad, SN: 12250707). Results are presented in table 7 as mean ± SEM of IL-6 concentrations (pg/ml). [0067] Table 7.

‘Differs significantly of control (p<0.05).

[0068] Table 7 shows that pulmonary administration of tocilizumab in doses of 0.1 to 100.0 mg/kg decreases overproduction of IL-6 in the lung of mammals suffering from IL-6 related respiratory disease in a dose-dependent manner.

Claims

Claims:

1. Tocilizumab for use in the treatment of an interleukin-6 related respiratory disease in a mammal by pulmonary administration in an amount of 0.1 to 100.0 mg per kg body weight of the mammal.

2. Tocilizumab as claimed in claim 1 , wherein the interleukin-6 related respiratory disease is selected from the group consisting of asthma, pneumonia, pulmonary edema, chronic obstructive pulmonary disease, and acute respiratory distress syndrome.

3. An aqueous pharmaceutical composition comprising tocilizumab and a pharmaceutically acceptable excipient for use in the treatment of an interleukin-6 related respiratory disease in a mammal by pulmonary administration of the composition in a dose containing 0.1 to 100.0 mg tocilizumab per kg body weight of the mammal.

4. The aqueous pharmaceutical composition as claimed in claim 3, wherein the interleukin- 6 related respiratory disease is selected from the group consisting of asthma, pneumonia, pulmonary edema, chronic obstructive pulmonary disease, and acute respiratory distress syndrome.

5. The aqueous pharmaceutical composition as claimed in any claims 3 to 4, wherein said composition is administered pulmonary in form of aerosol having particle size from 0.5 to 10.0 microns.

6. The aqueous pharmaceutical composition as claimed in any claims 3 to 5, wherein said excipient is selected from the group consisting of polysorbate 80, sucrose, disodium phosphate dodecahydrate, sodium dihydrogen phosphate dehydrate, L-histidine, L- histidine monohydrochloride, L-arginine, L-arginine hydrochloride, and L-methionine.

7. A method of treating an interleukin-6 related respiratory disease, which comprises a step of pulmonary administration of tocilizumab to a mammal in need thereof in an amount of 0.1 to 100.0 mg per kg body weight of the mammal.

8. The method as claimed in claim 7, wherein the interleukin-6 related respiratory disease is selected from the group consisting of asthma, pneumonia, pulmonary edema, chronic obstructive pulmonary disease, and acute respiratory distress syndrome.

9. The method as claimed in claim 7, wherein tocilizumab is administered one day or longer.

10. The method as claimed in claim 7, wherein tocilizumab is an active pharmaceutical ingredient of an aqueous pharmaceutical composition comprising tocilizumab and a pharmaceutically acceptable excipient.

11. The method as claimed in claim 10, wherein said composition is administered pulmonary in form of aerosol having particle size from 0.5 to 10.0 microns.

12. The method as claimed in claim 10, wherein said excipient is selected from the group consisting of polysorbate 80, sucrose, disodium phosphate dodecahydrate, sodium dihydrogen phosphate dehydrate, L-histidine, L-histidine monohydrochloride, L-arginine, L-arginine hydrochloride, and L-methionine.