Treatment of pneumonia

Abstract

To provide a composition to be used for treatment of pneumonia.SOLUTION: A composition including a mixture of empty liposomes is used for auxiliary treatment of pneumonia of a human patient, wherein the mixture of empty liposomes includes (a) first empty liposomes containing cholesterol, in which a volume of cholesterol is at least 45% to 55% (weight per weight), and (b) second empty liposomes containing sphingomyelin, which contain only the sphingomyelin as a lipid component, and the pneumonia is selected from severe pneumonia, community acquired pneumonia (CAP), hospitalization acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP) of the human patient.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a composition for use in the adjunctive treatment of pneumonia in a human patient, comprising a mixture of empty liposomes, the mixture comprising: (a) first empty liposomes comprising cholesterol, wherein the amount of cholesterol in the first empty liposomes is at least 30% (weight per weight); and (b) second empty liposomes comprising sphingomyelin. [Background technology]

[0002] The most challenging problem facing pneumonia and severe pneumonia today is the alarming increase in resistant bacteria and unacceptably high rates of treatment failure and mortality despite optimal care. Antimicrobial resistance poses a global threat to healthcare and the economy, and new treatment strategies and alternatives to antibiotics to overcome drug resistance are urgently needed (O'Neill, J., Review on Antimicrobial Resistance (2014); Bush, K. et al. Nature reviews. Microbiology 9, 894-896 (2011)). Retrospective reviews of the performance of new antibiotics have revealed that they alone are unable to significantly improve cure rates and / or reduce mortality (Azeredo da Silveira, S. and Perez, A. Expert Rev Anti Infect Ther 15, 973-975 (2017)).

[0003] Intensive care unit (ICU) patients with pneumonia continue to have a poor prognosis and may suffer long-term, difficult-to-treat consequences. For example, the mortality rate for adult ICU patients with severe pneumococcal pneumonia, caused by the most common pathogens Streptococcus pneumoniae, Staphylococcus aureus, and Pseudomonas aeruginosa, can still reach 40% despite optimal medical care and appropriate antibiotic therapy (Welte, T., et al., Thorax 67, 71-79 (2012); Torres, A. Community Acquired Infection 1, 1 (2017)). Streptococcus pneumoniae is the most frequently identified etiologic agent of community-acquired bacterial pneumonia, and its severe forms are associated with high morbidity and mortality (Blasi, F., et al. Clinical microbiology and infection: The official publication of the European Society of Clinical Microbiology and Infectious Diseases 18 Suppl 5, 7-14 (2012)). Despite preventive measures such as pneumococcal vaccines and current medical treatments (mainly antibiotic therapy, alone or in combination), there is a clear unmet medical need for additional treatment options (Vernatter, J. and Pirofski, L.A., Current opinion in infectious diseases 26, 277-283 (2013); Lucas, R. et al. Toxins 5, 1244-1260 (2013)).Staphylococcus aureus bacteremia has been persistently associated with a 20% 30-day mortality rate since the 1990s (Van Hal, SJ et al., Clinical microbiology reviews 25, 362-386 (2012)), and the mortality rate in ICU patients infected with methicillin-resistant Staphylococcus aureus (MRSA) is 50% higher than that expected for methicillin-susceptible Staphylococcus aureus (MSSA) infection (Hanberger, H. et al., International journal of antimicrobial agents 38, 331-335 (2011)). Multidrug-resistant Pseudomonas aeruginosa, which has become one of the most frequent causes of hospital-acquired and ventilator-associated pneumonia and is a major cause of morbidity in patients with cystic fibrosis or chronic obstructive pulmonary disease, has a mortality rate of over 35% (Ramirez-Estrada, S., et al. Infect Drug Resist 9, 7-18 (2016)).

[0004] Other bacteria, such as Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Moraxella catarrhalis, or Mycobacterium tuberculosis, can also cause respiratory infections and produce virulence factors that increase the severity of the disease and lead to complications.

[0005] Bacterial virulence factors promote colonization and proliferation in the host, disrupt tissue barriers, facilitate tissue penetration and bacterial dissemination, and disrupt the host's immune defense against pathogens (Los, FC, et al. MMBR 77, 173-207 (2013)). The pro- and anti-inflammatory cascades resulting from the release of bacterial toxins can lead to fatal complications.

[0006] Despite the best available treatments, many patients die from complications of severe pneumonia, which can occur several days after starting antibiotics, when tissues are already pathogen-free and lung disease progression has been eliminated. When patients with pneumonia are hospitalized, the infection develops over several days, the bacterial load peaks, and toxins are released in large quantities. For example, the Streptococcus pneumoniae toxin pneumolysin is not secreted into the extracellular medium but is released upon antibiotic-induced bacterial autolysis or lysis, resulting in its release in large quantities following antibiotic-induced bacterial lysis (Lucas, R. et al., Toxins 5, 1244-1260 (2013); Baumgartner, D. et al., BMC Microbiology 16, 154 (2016); Hirst, R.A. et al., Clin Exp Immunol 138, 195-201 (2004)). Notably, pneumolysin promotes disease progression, pneumococcal replication, and the development of invasive disease, playing a key role in severe and fatal complications. Indeed, the pathogenic effects of pneumolysin are multifactorial (Lucas, R. et al., Toxins 5, 1244-1260 (2013); Kadioglu, A., et al., Nature reviews. Microbiology 6, 288-301 (2008)). More specifically, pneumolysin causes widespread direct cell lysis and tissue damage. It affects the alveolar-capillary barrier, contributing to alveolar edema and hemorrhage, stimulating intraalveolar bacterial proliferation, and facilitating pneumococcus invasion into the pulmonary interstitium and bloodstream, thereby promoting pathogen invasiveness. Pneumolysin also alters the balance of local and systemic immune responses by inducing proinflammatory responses, exerting direct inhibitory effects on immune cells, and blocking complement, which prevents the initiation of the host immune response in the first hours of infection and the effective operation of the first line of immune defense against bacteria. Following the massive release of toxins that correlate with the bacteriolytic action of antibiotics, pneumolysin also plays an important role in the immunomodulatory side effects of antibiotic therapy, systemic inflammatory responses, and clinical deterioration.Toxin-mediated complications can occur and include respiratory failure, empyema, meningitis, arthritis, endocarditis, acute coronary syndrome, heart failure, osteomyelitis, hearing loss, epilepsy, hydrocephalus, and cognitive impairment.

[0007] Tailored empty liposomes, such as empty liposomes composed of cholesterol and / or sphingomyelin, and their use for the treatment of bacterial infections have recently been described, acting as traps for virulence factors such as bacterial toxins, enzymes, and toxic appendages (WO2013 / 186286, Henry BD et al., Nat Biotechnol 2015;33(1):81-88, Azeredo da Silveira, S and Perez, A, Expert Rev. Anti Infect. Ther. 2015;13(5):531-533). Summary of the Invention

[0008] A preferred composition of the present invention has shown surprisingly positive results in a first-in-human study for the treatment of pneumonia. Not only have the preferred composition of the present invention been shown to be safe and well-tolerated at all dose levels tested, but it has also provided consistent and encouraging efficacy data across a variety of clinical parameters. Based on this encouraging efficacy data and its possible mechanism of action, a second study is warranted, targeting hospitalized patients with suspected or confirmed infection, or signs of complications or severe illness, particularly those suffering from community-acquired pneumonia, hospital-acquired pneumonia, ventilator-associated pneumonia, intra-abdominal infections, skin and soft tissue infections, urinary tract infections, or bacteremia.

[0009] In the first study, a preferred composition of the present invention and standard antibiotic therapy were compared with placebo and standard antibiotic therapy in adult patients admitted to the intensive care unit (ICU) for severe community-acquired pneumococcal pneumonia. Two different doses of the preferred composition of the present invention were compared: a low dose (4 mg / kg-low dose) and a high dose (16 mg / kg-high dose). Results demonstrated a synergistic effect between the preferred composition of the present invention (designated CAL02) and antibiotic therapy.

[0010] As a result, preferred compositions of the present invention have been shown to synergistically capture and neutralize toxins released by a wide range of bacteria associated with severe infections in addition to antibiotic treatment in human patients. It is these toxins that are responsible for the development of severe and fatal complications. Therefore, preferred compositions of the present invention are first-in-class non-antibiotic liposomal drugs that are active against both Gram-positive and Gram-negative bacteria, including those that are multidrug-resistant. Preferred compositions of the present invention act regardless of the resistance profile of the target pathogen and do not induce the emergence of resistance.

[0011] Thus, in a first aspect, the present invention provides a composition comprising, preferably consisting of, a mixture of empty liposomes for use in the adjunctive treatment of pneumonia in a human patient, said mixture of empty liposomes comprising, preferably consisting of, (a) first empty liposomes comprising cholesterol, wherein the amount of cholesterol is at least 30% (weight per weight), and (b) second empty liposomes comprising sphingomyelin, preferably wherein the pneumonia is selected from community-acquired pneumonia (CAP), hospitalization-acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP).

[0012] In a further preferred embodiment, the pneumonia is severe pneumonia, preferably the severe pneumonia is selected from severe community-acquired pneumonia (sCAP), and even more preferably the severe pneumonia is severe community-acquired pneumococcal pneumonia (sCAPP) in human patients.

[0013] Further aspects and embodiments of the invention will become apparent as the description continues. [Brief explanation of the drawings]

[0014] [Figure 1A] Progression of the severity score SOFA (Figure 1A) and APACHE II (Figure 1B) from baseline (before administration) to day 8 in the placebo (diamond) group, the low-dose CAL02 (triangle) group, and the high-dose CAL02 (square) group. *p<0.05, **p<0.005 [Figure 1B] Progression of the severity score SOFA (Figure 1A) and APACHE II (Figure 1B) from baseline (before administration) to day 8 in the placebo (diamond) group, the low-dose CAL02 (triangle) group, and the high-dose CAL02 (square) group. *p<0.05, **p<0.005 [Figure 2A] Evolution of pharmacodynamic biomarkers from baseline (pre-dose) to day 8 in the placebo (diamond), CAL02 low-dose (triangle), and CAL02 high-dose (square) groups: C-reactive protein (CRP) (Figure 2A), procalcitonin (PCT) (Figure 2B), and interleukin-6 (Figure 2C). [Figure 2B] Evolution of pharmacodynamic biomarkers from baseline (pre-dose) to day 8 in the placebo (diamond), CAL02 low-dose (triangle), and CAL02 high-dose (square) groups: C-reactive protein (CRP) (Figure 2A), procalcitonin (PCT) (Figure 2B), and interleukin-6 (Figure 2C). [Figure 2C] Evolution of pharmacodynamic biomarkers from baseline (pre-dose) to day 8 in the placebo (diamond), CAL02 low-dose (triangle), and CAL02 high-dose (square) groups: C-reactive protein (CRP) (Figure 2A), procalcitonin (PCT) (Figure 2B), and interleukin-6 (Figure 2C). [Figure 3A] Time to ICU discharge ( Figure 3A ), p < 0.05, and time to hospital discharge ( Figure 3B ) in the placebo (diamond) and CAL02 high-dose (square) groups. [Figure 3B]Time to ICU discharge ( Figure 3A ), p < 0.05, and time to hospital discharge ( Figure 3B ) in the placebo (diamond) and CAL02 high-dose (square) groups. DETAILED DESCRIPTION OF THE INVENTION

[0015] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0016] As used herein, the term "about" means ±5%. For example, about 50% means 47.5% to 52.5%. Preferably, as used herein, the term "about" means ±3%. For example, about 50% means 48.5% to 51.5%.

[0017] When the terms "a" or "an" are used herein, unless otherwise indicated, they mean "at least one." In particular, the use of the terms "a" or "an" in relation to a single empty liposome, the first empty liposome, and the second empty liposome to describe empty liposomes and mixtures of empty liposomes according to the present invention typically and preferably refers to a single empty liposome(s), as well as a mixture of empty liposomes comprising the first empty liposome(s) and the second empty liposome(s).

[0018] All ranges of values disclosed herein are intended to refer to all values subsumed within that range, inclusive of the values defining the range. For clarity, for example, a value of 12 to 13 is intended to refer to the value 12 or 13, or to all values in the range 12 to 13.

[0019] The term "empty liposome" as used herein refers to a liposome, preferably an artificial liposome, having an average diameter of 20 nm to 10 μm, preferably 20 to 500 nm, more preferably 20 nm to 400 nm, and even more preferably 40 nm to 400 nm or 20 nm to 200 nm, and consisting of one or more phospholipid bilayers, and typically and preferably unilamellar and multilamellar vesicles, more preferably small unilamellar vesicles (SUVs). In a preferred embodiment, the term "empty liposome" as used herein typically and preferably refers to a liposome that does not incorporate a drug, typically and preferably refers to a liposome that does not incorporate a pharmaceutical drug. As used herein and referring to the empty liposomes of the present invention, "incorporated" typically and preferably means encapsulated within the cavity of the liposome, within the latent bilayer of the liposome, or as part of the membrane layer of the liposome. In another preferred embodiment, the term "empty liposome" as used herein typically and preferably refers to a liposome consisting of sphingomyelin and cholesterol according to the present invention, or consisting of sphingomyelin, and further comprising only a water-soluble inorganic compound and / or a water-soluble organic molecule, typically and preferably the water-soluble inorganic compound and / or the water-soluble organic molecule originating from the synthesis of the empty liposome of the present invention, typically and preferably the water-soluble inorganic compound is an inorganic salt preferably selected from NaCl, KCl, and MgCl, and the water-soluble organic molecule is a buffer, preferably selected from glucose and HEPES. Typically and preferably, the water-soluble inorganic compound and / or the water-soluble organic molecule are incorporated into the empty liposome of the present invention due to their presence during the production of the empty liposome of the present invention. In another preferred embodiment, the term "empty liposome" as used herein typically and preferably refers to a liposome consisting of sphingomyelin and cholesterol according to the present invention, or consisting of sphingomyelin, in which the empty liposome does not contain an antioxidant.In a further preferred embodiment, the term "empty liposome" as used herein typically and preferably refers to a liposome consisting of sphingomyelin and cholesterol according to the present invention, or consisting of sphingomyelin, and further comprising only water-soluble inorganic compounds and / or water-soluble organic molecules, typically and preferably the water-soluble inorganic compounds and / or water-soluble organic molecules originating from the synthesis of the empty liposome according to the present invention, and typically and preferably the water-soluble inorganic compounds are inorganic salts preferably selected from NaCl, KCl, MgCl2, and the water-soluble organic molecules are buffers, preferably the water-soluble organic molecules are selected from glucose and HEPES, and the empty liposomes consisting of sphingomyelin and cholesterol according to the present invention or consisting of sphingomyelin do not contain antioxidants.

[0020] As used herein, the terms "treat," "treatment," or "therapy" refer to a means of obtaining a desired physiological effect. The effect may be therapeutic in terms of partially or completely curing a disease or condition and / or symptoms caused by the disease or condition. The term refers to inhibiting a disease or condition, i.e., preventing its occurrence, or ameliorating a disease or condition, i.e., causing regression of the disease or condition.

[0021] As used herein, the term "for use" as used in "a composition for use in the treatment of a disease" is intended to also disclose the corresponding method of treatment and the use of the formulation for the manufacture of a medicament for the treatment of the corresponding disease."

[0022] The term "pneumonia" as used herein shall encompass "community-acquired pneumonia" (CAP), "hospital-acquired pneumonia" (HAP) or "ventilator-associated pneumonia" (VAP).

[0023] The term "community-acquired pneumonia" or "CAP" is known to those skilled in the art; see, e.g., the IDSA / ATS Guidelines for CAP in Adults (CID 2007:44(Suppl 2)S27). In particular, this term refers to pneumonia acquired outside of a hospital.

[0024] The term "hospital-acquired pneumonia (HAP)" refers to pneumonia acquired during or after hospitalization for another illness or procedure, with onset at least 48 to 72 hours after admission.

[0025] As defined herein, "ventilator-associated pneumonia (VAP)" is pneumonia that develops after 48 hours or more of mechanical ventilation and is characterized by invasion of the lower airways and lung parenchyma by microorganisms. VAP is a potentially serious condition.

[0026] Pneumonia is caused by infection with a wide range of microorganisms, including bacteria such as Streptococcus pneumoniae, Haemophilus influenzae, Legionella pneumophilia, Staphylococcus aureus and Pseudomonas aeruginosa, and gram-negative bacteria such as Pseudomonas aeruginosa and Serratia marcescens for CAP, and Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Stenotrophomonas maltophilia, Acinetobacter species, and Haemophilus influenzae for HAP (Cilloniz et al., Thorax. 2011 Apr;66(4):340-6 and Jones RN. Clin Infect Dis 2010 Aug;51(Suppl 1):S81-7).

[0027] The term "severe community-acquired pneumonia" or "sCAP" is known to those skilled in the art. In particular, the term "severe community-acquired pneumonia" or "sCAP" refers to a subgroup of community-acquired pneumonia patients who require intensive care. The Infectious Diseases Society of America (IDSA) and the American Thoracic Society (ATS) have published guidelines for the management of CAP, including a definition of sCAP (see Mandell et al., 2007, Infectious Diseases Society of America / American Thoracic Society Consensus Guidelines on the Management of Community-Acquired Pneumonia in Adults, Clin. Inf. Dis. 2007:44:S27-72 (Suppl 2), Table 4). According to the IDSA / ATS guidelines, sCAP is defined as CAP requiring intensive care. Admission to an intensive care unit is recommended if a CAP patient exhibits one or both of the two major criteria or if the three minor criteria listed in Example 1 and affected in this study are present.

[0028] Thus, in a first aspect, the present invention provides a composition comprising, preferably consisting of, a mixture of empty liposomes for use in the adjunctive treatment of pneumonia in a human patient, the mixture of empty liposomes comprising, preferably consisting of, (a) first empty liposomes comprising cholesterol, wherein the amount of cholesterol is at least 30% (weight per weight), and (b) second empty liposomes comprising sphingomyelin, preferably wherein the pneumonia is selected from community-acquired pneumonia (CAP), hospital-acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP), more preferably wherein the pneumonia is community-acquired pneumonia (CAP), and even more preferably wherein the pneumonia is community-acquired pneumococcal pneumonia (CAPP) in a human patient.

[0029] In another aspect, the present invention provides a composition comprising, preferably consisting of, a mixture of empty liposomes for use in the adjunctive treatment of pneumonia in a human patient, wherein the mixture of empty liposomes comprises, preferably consists of, (a) first empty liposomes comprising cholesterol, wherein the amount of cholesterol is at least 30% (weight per weight), and (b) second empty liposomes comprising sphingomyelin, preferably wherein the pneumonia is severe pneumonia, preferably wherein the severe pneumonia is selected from severe community-acquired pneumonia (sCAP), hospital-acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP), more preferably wherein the severe pneumonia is severe community-acquired pneumonia (sCAP), and even more preferably wherein the severe pneumonia is severe community-acquired pneumococcal pneumonia (sCAPP) in a human patient.

[0030] In a preferred embodiment, the second empty liposome (b) comprises only the sphingomyelin as a lipid component.

[0031] In a preferred embodiment, the second empty liposome (b) consists of sphingomyelin.

[0032] In a preferred embodiment, the amount of cholesterol in the first empty liposome (a) is 30% to 70% (weight per weight), preferably, the amount of cholesterol in the first empty liposome (a) is 35% to 60% (weight per weight).

[0033] In a preferred embodiment, the amount of cholesterol in the empty liposome (a) is 45% to 55% (weight per weight), preferably the amount of cholesterol in the empty liposome (a) is about 50% (weight per weight).

[0034] In a preferred embodiment, the first empty liposome (a) consists of cholesterol and sphingomyelin, and preferably the amount of cholesterol in the empty liposome (a) is 45% to 55% (weight per weight), and more preferably the amount of cholesterol in the empty liposome (a) is about 50% (weight per weight).

[0035] In a preferred embodiment, the first empty liposome (a) contains only the cholesterol and the sphingomyelin as lipid components.

[0036] In a preferred embodiment, the amount of cholesterol in the empty liposome (a) is 45% to 55% (weight per weight), preferably the amount of cholesterol in the empty liposome (a) is about 50% (weight per weight), and the second empty liposome (b) consists of sphingomyelin.

[0037] In a preferred embodiment, the first empty liposome (a) consists of cholesterol and sphingomyelin, the amount of cholesterol in the empty liposome (a) is 45% to 55% (weight per weight), preferably the amount of cholesterol in the empty liposome (a) is about 50% (weight / weight), and the second empty liposome (b) consists of sphingomyelin.

[0038] In a preferred embodiment, the first empty liposome (a) consists of cholesterol and sphingomyelin, the amount of cholesterol in the empty liposome (a) is 45% to 55% (weight per weight), preferably the amount of cholesterol in the empty liposome (a) is about 50% (weight per weight), the first empty liposome (a) contains only cholesterol and sphingomyelin as lipid components, and the second empty liposome (b) consists of sphingomyelin.

[0039] In a preferred embodiment, the mixture of empty liposomes comprises at least 20% (weight per weight) of the first(a) and second(b) empty liposomes, preferably the mixture of empty liposomes comprises at least 30% (weight per weight) of the first(a) and second(b) empty liposomes.

[0040] In a preferred embodiment, said mixture of empty liposomes comprises at least 40% (weight per weight) of said first(a) and said second(b) empty liposomes.

[0041] In a preferred embodiment, the first empty liposome (a) consists of cholesterol and sphingomyelin, more preferably the amount of cholesterol in the empty liposome (a) is about 50% (weight per weight), and the mixture of empty liposomes comprises at least 40%, preferably at least 45% (weight per weight) of the first (a) and the second (b) empty liposomes.

[0042] In a preferred embodiment, the first empty liposome (a) consists of a 1:1 (weight per weight - w / w) mixture of the first empty liposome and the second liposome, the first empty liposome being composed of cholesterol and sphingomyelin in a 1:1 weight ratio (1:1 w / w, 35:65 molar ratio), and the second empty liposome being composed of sphingomyelin alone.

[0043] In a preferred embodiment, the first empty liposome (a) consists of a 1:1 (weight per weight - w / w) mixture of the first empty liposome and the second liposome, the first empty liposome being composed of cholesterol and sphingomyelin in a 1:1 weight ratio (1:1 w / w, 35:65 molar ratio), and the second empty liposome being composed solely of sphingomyelin, the first empty liposome (a) comprising only the cholesterol and the sphingomyelin as lipid components, and the second empty liposome (b) comprising only the sphingomyelin as lipid component.

[0044] In a preferred embodiment, the first empty liposomes have an average diameter of about 130 nm and the second empty liposomes have an average diameter of about 90 nm.

[0045] In a preferred embodiment, the composition is for use in the adjunctive treatment of pneumonia.

[0046] In a preferred embodiment, the pneumonia is community-acquired pneumonia (CAP). In a preferred embodiment, the pneumonia is hospital-acquired pneumonia (HAP). In a preferred embodiment, the pneumonia is ventilator-associated pneumonia (VAP).

[0047] In a preferred embodiment, the composition is for use in the adjunctive treatment of pneumonia, wherein the pneumonia is severe pneumonia.

[0048] In a preferred embodiment, the composition is for use in the adjunctive treatment of pneumonia, wherein the pneumonia is community-acquired pneumonia (CAP) in a human patient. In a preferred embodiment, the pneumonia is severe pneumonia, wherein the severe pneumonia is severe community-acquired pneumococcal pneumonia (sCAPP).

[0049] In a preferred embodiment, the composition is for use in the adjunctive treatment of pneumonia, wherein the pneumonia is hospital-acquired pneumonia (HAP) in a human patient.

[0050] In a preferred embodiment, the composition is for use in the adjunctive treatment of pneumonia, wherein the pneumonia is ventilator-associated pneumonia (HAP) in a human patient.

[0051] In a preferred embodiment, the composition is for use in the adjunctive treatment of pneumonia, wherein the pneumonia is community-acquired pneumococcal pneumonia (CAPP) in a human patient.

[0052] In a preferred embodiment, the severe pneumonia is severe community-acquired pneumonia (sCAP), preferably the severe pneumonia is severe community-acquired pneumococcal pneumonia (sCAPP) in a human patient.

[0053] In preferred embodiments, the pneumonia is caused by Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecium, Legionella pneumophila, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Serratia marcescens, Stenotrophomonas maltophilia, Moraxella catarrhalis, or Mycobacterium tuberculosis.

[0054] In a preferred embodiment, the pneumonia is severe pneumonia caused by Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecium, Legionella pneumophila, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Serratia marcescens, Stenotrophomonas maltophilia, Moraxella catarrhalis, or Mycobacterium tuberculosis.

[0055] In preferred embodiments, the pneumonia, preferably the severe pneumonia, is caused by Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecium, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Moraxella catarrhalis, or Mycobacterium tuberculosis.

[0056] In preferred embodiments, the pneumonia, preferably the severe pneumonia, is caused by Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecium, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Moraxella catarrhalis, or Mycobacterium tuberculosis.

[0057] In preferred embodiments, the pneumonia is caused by Streptococcus pneumoniae, Staphylococcus aureus, Klebsiella pneumoniae, or Pseudomonas aeruginosa.

[0058] In a preferred embodiment, the pneumonia, preferably the severe pneumonia, is caused by Streptococcus pneumoniae, Staphylococcus aureus, Klebsiella pneumoniae, or Pseudomonas aeruginosa.

[0059] In a preferred embodiment, the pneumonia is caused by Streptococcus pneumoniae.

[0060] In a preferred embodiment, the pneumonia is severe pneumonia caused by Streptococcus pneumoniae.

[0061] In a preferred embodiment, the pneumonia, preferably the severe pneumonia, is severe community-acquired pneumonia (sCAP) caused by Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecium, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Moraxella catarrhalis, or Mycobacterium tuberculosis, preferably, the sCAP is caused by Streptococcus pneumoniae, Staphylococcus aureus, or Pseudomonas aeruginosa, and more preferably, the sCAP is caused by Streptococcus pneumoniae.

[0062] In a preferred embodiment, the pneumonia, preferably the severe pneumonia, is hospital-acquired pneumonia (HAP) caused by Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Serratia marcescens, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecium, Acinetobacter baumannii, Bordetella pertussis, Moraxella catarrhalis, or Mycobacterium tuberculosis, preferably the HAP is caused by Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, or Pseudomonas aeruginosa.

[0063] In a preferred embodiment, the pneumonia, preferably the severe pneumonia, is hospital-acquired pneumonia (HAP) caused by Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Serratia marcescens, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecium, Acinetobacter baumannii, Bordetella pertussis, Moraxella catarrhalis, or Mycobacterium tuberculosis, pertussis, Moraxella catarrhalis, or Mycobacterium tuberculosis, preferably, the HAP is caused by Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, or Pseudomonas aeruginosa.

[0064] In a preferred embodiment, the pneumonia, preferably the severe pneumonia, is severe community-acquired pneumonia (sCAP), and the severe community-acquired pneumonia (sCAP) is severe community-acquired bacterial pneumonia, preferably severe community-acquired pneumococcal pneumonia (sCAPP) in a human patient, and more preferably, the sCAPP is sCAPP caused by Streptococcus pneumoniae.

[0065] In a preferred embodiment, the pneumonia, preferably the severe pneumonia, is caused by a bacterial infection, and the bacterial infection is Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Moraxella catarrhalis, or Mycobacterium tuberculosis; preferably, the bacterial infection is an infection by Streptococcus pneumoniae, Staphylococcus aureus, or Pseudomonas aeruginosa; more preferably, the bacterial infection is an infection by Streptococcus pneumoniae.

[0066] In a preferred embodiment, the pneumonia, preferably the severe pneumonia, is severe community-acquired pneumonia (sCAP) caused by a bacterial infection, wherein the bacterial infection is Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Moraxella catarrhalis, or Mycobacterium tuberculosis, preferably the bacterial infection is an infection caused by Streptococcus pneumoniae, Staphylococcus aureus, or Pseudomonas aeruginosa, more preferably the bacterial infection is an infection caused by Streptococcus pneumoniae.

[0067] In a preferred embodiment, the pneumonia, preferably the severe pneumonia, is severe community-acquired pneumonia (sCAP), the sCAP is sCAP caused by a bacterial infection, the bacterial infection is a pneumococcal infection, and the pneumococcal infection is an infection caused by Streptococcus pneumoniae.

[0068] In a preferred embodiment, the treatment is adjunctive to antibiotic therapy, preferably standard antibiotic therapy.

[0069] In a preferred embodiment, the antibiotic therapy, preferably the standard antibiotic therapy, is selected from the group consisting of ceftriaxone, spiramycin, amoxicillin, amoxicillin / clavulanic acid, gentamicin, piperacillin / tazobactam, cefuroxime, penicillin, azithromycin, clarithromycin, erythromycin, doxycycline, cefotaxime, ampicillin, ertapenem, cefepime, imipenem, meropenem, cipro Preferably, the antibiotic of the antibiotic therapy, preferably the antibiotic of the standard antibiotic therapy, is selected from ceftriaxone, spiramycin, amoxicillin, gentamicin, levofloxacin, piperacillin / tazobactam, amoxicillin / clavulanic acid, cefuroxime, and penicillin.

[0070] In a preferred embodiment, the antibiotic therapy, preferably a standard antibiotic therapy, is an intravenous (IV) or oral antibiotic therapy, preferably a standard intravenous (IV) or oral antibiotic therapy.

[0071] In a preferred embodiment, the composition is in the form of a solution for intravenous administration, preferably intravenous infusion, and comprises 10 to 40 grams of the empty liposome mixture per liter of solution, preferably 10 to 20 grams of the empty liposome mixture per liter of solution.

[0072] In a preferred embodiment, the infusion time for the intravenous administration is up to 3 hours, and preferably, the infusion time for the intravenous administration is 10 minutes to 2 hours.

[0073] In a preferred embodiment, the composition is administered in at least two doses, a first dose and a second dose, and the interval between the first dose and the second dose is 6 to 96 hours, preferably 12 to 72 hours, more preferably 24 to 48 hours, and even more preferably 24 or 48 hours.

[0074] In a preferred embodiment, the composition is administered in two to four doses, preferably two doses over a 12 to 72 hour period, more preferably two doses over a 24 to 48 hour period, preferably 24 or 48 hour intervals.

[0075] In a preferred embodiment, the composition is administered to the human patient in at least two doses, a first dose and a second dose, the interval between the first dose and the second dose being 20 to 28 hours, preferably 24 hours.

[0076] In a preferred embodiment, each of the doses is 1 mg / kg to 64 mg / kg, preferably 2 mg / kg to 32 mg / kg, more preferably 3 to 25 mg / kg, and even more preferably 4 mg / kg to 16 mg / kg.

[0077] In a preferred embodiment, each of the doses is 2 mg / kg to 8 mg / kg, preferably 2 mg / kg to 6 mg / kg, more preferably 3 to 5 mg / kg, and even more preferably 4 mg / kg.

[0078] In a preferred embodiment, each of said doses is 10 mg / kg to 22 mg / kg, preferably 12 mg / kg to 20 mg / kg, more preferably 14 mg / kg to 18 mg / kg, and even more preferably 16 mg / kg.

[0079] In a preferred embodiment, the composition is administered in the form of an intravenous solution.

[0080] In a preferred embodiment, the composition is administered to the human patient in at least two doses, preferably a first dose and a second dose, with an interval of 20 to 48 hours, preferably 24 hours, between the administration of the first dose and the administration of the second dose, and the composition is in the form of a solution for intravenous administration.

[0081] In a preferred embodiment, the composition is administered in at least two doses, the first of which is administered within 12 hours of diagnosis of said pneumonia, preferably said CAP or sCAP, more preferably said sCAPP.

[0082] In a preferred embodiment, the composition is administered in at least two doses, with the first dose being administered within 24 hours of initiating the antibiotic therapy, preferably IV antibiotic therapy.

[0083] In a preferred embodiment, the composition is administered in at least two doses, the first dose being administered within 60 hours after the initiation of the oral antibiotic therapy.

[0084] In a preferred embodiment, the pneumonia requires hospitalization, preferably in the intensive care unit (ICU) of a hospital.

[0085] In a preferred embodiment, the severe pneumonia, more preferably the sCAP, requires hospitalization.

[0086] In a preferred embodiment, said pneumonia, preferably said severe pneumonia, more preferably said sCAP, requires admission to an intensive care unit (ICU), preferably in an intensive care unit (ICU) of a hospital.

[0087] In a preferred embodiment, the pneumonia, preferably the severe pneumonia, more preferably the sCAP, requires hospitalization of the patient, and the treatment is adjunctive to antibiotic therapy, preferably standard antibiotic therapy, and the adjunctive treatment shortens the duration of the hospitalization compared to hospitalization in the absence of such treatment.

[0088] In a preferred embodiment, the adjunctive treatment reduces hospital stay by at least 1 day, preferably 2 days, more preferably 3 days, even more preferably 4 days, even more preferably 5 days, even more preferably 6 days, even more preferably 7 days, even more preferably 8 days, and even more preferably 9 days.

[0089] In a preferred embodiment, the hospital stay is a maximum of two weeks.

[0090] In a preferred embodiment, the pneumonia, preferably the severe pneumonia, more preferably the sCAP, requires admission to an intensive care unit (ICU), and the treatment is adjunctive to antibiotic therapy, preferably standard antibiotic therapy, and the adjunctive treatment shortens the length of stay in the intensive care unit (ICU) compared to admission to the intensive care unit (ICU) in the absence of such treatment.

[0091] In a preferred embodiment, the reduction in length of stay in the intensive care unit (ICU) is at least 1 day, preferably 2 days, more preferably 3 days, even more preferably 4 days, even more preferably 5 days, even more preferably 6 days, and even more preferably 7 days.

[0092] In a preferred embodiment, the length of stay in the intensive care unit (ICU) is a maximum of one week.

[0093] In a preferred embodiment, the pneumonia, preferably the severe pneumonia, more preferably the sCAP, is cured in a shorter time than if such adjunctive treatment were not administered, preferably compared to if such treatment were not administered adjunctive to the antibiotic therapy, preferably the standard antibiotic therapy.

[0094] In a preferred embodiment, the shorter healing time is at least 1 day short, preferably 2 days short, or more preferably at least 3 days short.

[0095] In a preferred embodiment, said cure in a shorter time is manifested by normalization of organ function that is at least 1 day shorter, preferably 2 days shorter, or even more preferably at least 3 days shorter, compared to the absence of such adjunctive treatment, preferably the absence of such treatment adjunctive to said antibiotic therapy, preferably said standard antibiotic therapy.

[0096] In a preferred embodiment, said healing in a shorter time is reflected by a normalization of blood inflammatory markers, preferably CRP and / or PCT and / or IL-6, that is at least 1 day shorter, preferably 2 days shorter, or even more preferably at least 3 days shorter, compared to the absence of such adjunctive treatment, preferably the absence of such treatment adjunctive to said antibiotic therapy, preferably said standard antibiotic therapy.

[0097] In a preferred embodiment, said cure in a shorter time is manifested by normalization of organ function and normalization of blood inflammatory markers, preferably CRP and / or PCT and / or IL-6, that is at least 1 day shorter, preferably 2 days shorter or even more preferably at least 3 days shorter compared to the absence of such adjunctive treatment, preferably the absence of such treatment adjunctive to said antibiotic therapy, preferably said standard antibiotic therapy.

[0098] In a preferred embodiment, said cure in a shorter time is indicated by a decrease in the SOFA score of at least 1 day shorter, preferably 2 days shorter, or even more preferably at least 3 days shorter compared to the absence of such adjunctive treatment, preferably the absence of such treatment adjunctive to said antibiotic therapy, preferably said standard antibiotic therapy.

[0099] In a preferred embodiment, said cure in a shorter time is indicated by a decrease in APACHE II score of at least 1 day shorter, preferably 2 days shorter, or more preferably at least 3 days shorter, compared to the absence of such adjunctive treatment, preferably the absence of such treatment adjunctive to said antibiotic therapy, preferably the standard antibiotic therapy.

[0100] In a preferred embodiment, said cure in a shorter time is reflected by a decrease in APACHE II score and SOFA score of at least 1 day shorter, preferably 2 days shorter, or more preferably at least 3 days shorter, compared to the absence of such adjunctive treatment, preferably the absence of such treatment adjunctive to said antibiotic therapy, preferably said standard antibiotic therapy.

[0101] In a preferred embodiment, said treating said pneumonia, preferably said severe pneumonia, more preferably said sCAP, even more preferably said sCAPP reduces said patient's SOFA score by at least 40% 8 days after the first dose of said composition, preferably 7 days after the first dose of said composition, even more preferably 6 days after the first dose of said composition, even more preferably 5 days after the first dose of said composition.

[0102] In a preferred embodiment, said treating said pneumonia, preferably said severe pneumonia, more preferably said sCAP, even more preferably said sCAPP reduces said patient's APACHE II score by at least 20% 8 days after the first dose of said composition, preferably 6 days after the first dose of said composition, even more preferably 5 days after the first dose of said composition, and even more preferably 4 days after the first dose of said composition. [Example]

[0103] Liposomes: Sphingomyelin from egg yolk (CAS No. 85187-10-6) was purchased from Sigma (S0756), Avanti Polar Lipids (860061) or Lipoid GmbH. Cholesterol from wool grease (CAS No. 57-88-5) was purchased from Sigma (C-8667), Avanti Polar Lipids (70000) or Dishman Netherlands BV. According to the present invention, the sphingomyelin and cholesterol contained in or consisting of the inventive mixture of empty liposomes can be obtained from the above natural sources or by chemical synthesis.

[0104] Liposome preparation: Unilamellar sphingomyelin:cholesterol (35:65 molar ratio) and sphingomyelin-only (100%) liposomes were prepared using sonication or microfluidization (e.g., high-pressure homogenization) or following a hydration, extrusion, and diafiltration process protocol.

[0105] Sonication: Lipids were individually dissolved in chloroform at a concentration of 1 mg / ml and stored at -20°C. For liposome preparation, chloroform solutions of individual lipids were routinely mixed in the appropriate proportions to produce a final solution of 50–500 μl. The chloroform was allowed to completely evaporate at 60°C for 20–50 min. 50 μl or 100 μl of Tyrode's buffer (140 mM NaCl, 5 mM KCl, 1 mM MgCl, 10 mM glucose, 10 mM HEPES, pH = 7.4) containing 2.5 mM CaCl was added to the tube containing the dried lipid film and vortexed vigorously. The lipid suspension was incubated in an Eppendorf thermomixer at 45°C for 20–30 min with vigorous shaking. To produce liposomes, the final lipid suspension was sonicated for 3 x 5 s at 6°C at 70% power in a Bandelin Sonopuls sonicator. The liposome preparation was left at 6°C for at least 1 hour before use in experiments.

[0106] Hydration, Extrusion, and Diafiltration Process Protocol: Alternatively, each liposome formulation was produced by ethanol hydration and extrusion. Lipids were dissolved separately in ethanol and t-butanol and mixed at elevated temperatures (approximately 55 °C). The lipid solution was then added to PBS buffer (sodium chloride, monosodium phosphate, dihydrate, and disodium phosphate, dihydrate, dissolved in water for injection during mixing, adjusted to pH 7.0-7.4 with either hydrochloric acid (HCl) or sodium hydroxide (NaOH), as needed, and filtered through a 0.2 µm filter) and mixed at elevated temperatures (approximately 65 °C) for approximately 30 minutes. The resulting process fluid was then repeatedly extruded through a series of polycarbonate track-etched membranes at elevated pressures and temperatures (approximately 65 °C) until the desired particle size, as measured by dynamic light scattering, was achieved (example extruder: LIPEX® extruder). The resulting process fluid was then concentrated approximately two-fold using a 100,000 molecular weight cutoff hollow fiber cartridge and then diafiltered against approximately 10 volume changes of PBS buffer solution to remove ethanol and t-butanol. At the end of diafiltration, the process fluid was concentrated approximately 30% and then diluted to the target lipid concentration. Prior to dilution, the process fluid was filtered through a 0.2 μm sterilizing-grade filter to remove any large liposomes that could clog the filter during sterile filtration. The process fluid was then diluted to a target of 40 mg / mL total lipid using PBS buffer. The final formulation was aseptically filtered through two 0.2 μm sterilizing-grade filters in series and aseptically filled into glass vials.

[0107] The concentrations of individual lipids in liposomes are always given as a weight-per-weight ratio. For liposomes containing sphingomyelin and cholesterol, a 1:1 (weight-per-weight) ratio corresponds to 50% (weight-per-weight) or a 35:65 molar ratio. Specifications are shown in Table 1. JPEG2025118655000001.jpg48170

[0108] CURB-65 score: The CURB-65, also known as the CURB criteria, is a clinical prediction rule well known to those skilled in the art that has been validated to predict mortality in community-acquired pneumonia (Lim WS, et al. (2003) Thorax 58(5):377-82). The CURB-65 is recommended by the British Thoracic Society for assessing the severity of pneumonia (British Thoracic Society Standards of Care Committee (2001). "BTS Guidelines for the Management of Community-Acquired Pneumonia in Adults". Thorax. 56. Suppl 4:IV1-64).

[0109] The score is an acronym for each risk factor measured. Each risk factor has a score of 1, with a maximum score of 5. -First-onset confusion (defined as an AMTS of 8 or less) Blood urea nitrogen greater than 7 mmol / l (19 mg / dL) Respiratory rate of 30 breaths per minute or more Systolic blood pressure less than 90mmHg or diastolic blood pressure less than 60mmHg -Over 65 years old.

[0110] APACHE II: APACHE II is an acronym for Acute Physiology and Chronic Health Evaluation and is calculated according to Table 2 (Knaus WA et al. APACHE II: a severity of disease classification system. Crit Care Med. 1985 Oct;13(10):818-29): JPEG2025118655000002.jpg255144JPEG2025118655000003.jpg255147JPEG2025118655000004.jpg238170

[0111] The APACHE II score is intended to provide an estimate of mortality for a patient or group of patients based on the total score obtained. Mortality interpretation of the score is shown in Table 3. JPEG2025118655000005.jpg85170

[0112] Sofa: The SOFA score is an acronym for Sequential Organ Failure Assessment and is calculated according to Table 4 (S. Vosylius, J. Sipylaite and J. Ivaskevicius, Croat Med J, 45 (2004), 715-20). JPEG2025118655000006.jpg182170

[0113] Example 1 Treatment of patients with severe community-acquired pneumonia caused by Streptococcus pneumoniae In a subsequently reported study, a highly preferred mixture of empty liposomes of the present invention, designated CAL02, was administered intravenously (IV) as an adjunctive treatment to standard antibiotic therapy in patients with severe community-acquired pneumonia (CAP) due to Streptococcus pneumoniae requiring intensive care unit (ICU) care. The highly preferred mixture of empty liposomes of the present invention (CAL02) consisted of a 1:1 (weight per weight - w / w) mixture of the first empty liposomes and the second liposomes, where the first empty liposomes were composed of sphingomyelin and cholesterol in a 1:1 weight ratio (1:1 w / w, 35:65 molar ratio) and the second empty liposomes were composed solely of sphingomyelin.

[0114] All 19 patients (male and female) in this study received standard-of-care antibiotic therapy according to standard practice (Mandell et al., IDSA / ATS Guidelines for CAP in Adults, CID 2007:44(Suppl 2), S27-S72) and additionally received CAL02 or placebo (physiological 0.9% NaCl solution). Two dose levels of CAL02 were tested: 4 mg / kg (low dose) and 16 mg / kg (high dose).

[0115] Each patient received two infusions of CAL02 or placebo (24 or 48 hours between these doses) immediately after the diagnosis of severe disease (within 12 hours after the diagnosis of severe CAP or severe community-acquired pneumococcal pneumonia (CAPP)) and early in antibiotic treatment (within 24 hours after the start of IV antibiotic treatment (within 60 hours for oral antibiotics)). Patients were followed for a total of 29 days.

[0116] In this study, all patients received standard-of-care antibiotic therapy according to local guidelines, with antibiotics alone or in combination, and the dose and frequency of the standard-of-care antibiotic therapy selected by the investigator followed the approved treatment labeling in each country.

[0117] The inclusion criteria for this study were, inter alia: 1. Includes men or women aged 18 to 80 years, weighing 40 to 140 kg; 2. Patients diagnosed with severe CAP, with new pulmonary infiltrates found on chest X-ray, newly acquired respiratory symptoms (cough, sputum production), and abnormal breath sounds and crackles on auscultation. 3. CURB-65 score ≥3 for patients ≥65 years old and CURB-65 ≥2 for patients <65 years old 4. CAPP is diagnosed with severe pneumonia based on the major or minor severity criteria listed below, requiring intensive care unit (ICU) admission (screening can be performed in the emergency department).

[0118] Key criteria: Patients who meet at least one of the two following key severity criteria for CAP: i. Receiving invasive mechanical ventilatory support ii. After adequate fluid resuscitation, receive vasopressor therapy at therapeutic doses (i.e., dopamine >5 mg / kg / min or any dose of epinephrine, norepinephrine, phenylephrine, or vasopressin) to maintain or attempt to maintain a systolic blood pressure >90 mmHg (or mean arterial pressure >70 mmHg) for at least 2 hours.

[0119] Minor Criteria: Patients who meet three or more of the following minor severity criteria for CAP: i.Respiration rate ≧30 breaths / min ii.PaO2 / FiO2 ratio≦250mmHg iii.Multilobular infiltrate iv. Confusion / disorientation (must be documented prior to use of sedatives or other new psychotropic medications) v.Urea>7mM(>40mg / dL) vi. Leukopenia (white blood cell count <4'000 cells / mm 3 ) vii. Thrombocytopenia (platelet count <100'000 cells / mm 3 ) viii. Hypothermia (core body temperature <36°C) ix. Systolic blood pressure <90mmHg or mean arterial pressure <70mmHg and received fluid resuscitation of ≥40mL / kg for at least 2 hours

[0120] Research results Five patients were randomly assigned to the placebo group, 11 to the high-dose CAL02 group, and 3 to the low-dose CAL02 group. One patient in the high-dose group was removed before unblinding due to a violation of a key eligibility criterion. All patients received antibiotic treatment or combinations of antibiotics deemed appropriate for the patient. Overall, ceftriaxone, spiramycin, amoxicillin, gentamicin, levofloxacin, piperacillin / tazobactam, amoxicillin / clavulanate, cefuroxime, and penicillin were used in this study population.

[0121] The mean APACHE II score at treatment was 21.3, and SOFA score was 7.4. Baseline characteristics were homogeneous across groups. However, the CAL02 group proved to be slightly more severely ill than the placebo group, with APACHE II scores of 22.1 and 25.3 in the high-dose and low-dose CAL02 groups, respectively, compared with 17.4 in the placebo group. All three low-dose patients were intubated. Baseline characteristics are summarized in Table 5. JPEG2025118655000007.jpg168170

[0122] Nineteen patients were infected with S. pneumoniae. One patient in the low-dose cohort was also infected with Escherichia coli. One patient in the low-dose cohort was also infected with Klebsiella pneumoniae. One patient in the high-dose cohort was also infected with Enterococcus faecium. Three patients (one in the low-dose and two in the high-dose cohort) were infected with Staphylococcus aureus.

[0123] One patient died in each group, with mortality rates of 20% in the placebo group, 10% in the high-dose CAL02 group, and 33% in the low-dose CAL02 group.

[0124] Time of Cure (TOC) visits were predefined at day 8 (early TOC), days 15 through 22, and at the end of study (EOS). The TOC is a medical visit where the determination of (clinical) cure and therefore the efficacy of treatment is assessed.

[0125] Clinical cure in this study corresponded to when the patient showed resolution of pneumonia signs and symptoms, when standard laboratory data returned to normal, when fever subsided (without antipyretics for at least 24 hours), and when the patient showed improvement or clean Rx films and no longer required antibiotic treatment.

[0126] At the TOC visit, clinical cure (or clinical outcome) was assessed as follows: a) Cure: Complete resolution of signs and symptoms of pneumonia present at baseline, no new symptoms or complications attributable to pneumonia. b) Failure: persistence / progression of baseline signs and symptoms of pneumonia, or baseline radiological abnormalities at least 2 days after treatment, or development of new pulmonary or extrapulmonary clinical findings consistent with active infection, or development of new pulmonary or extrapulmonary infection requiring antimicrobial therapy other than or in addition to IMP, or death from pneumonia. c) Unknown: Extenuating circumstances preclude classification as one of the above.

[0127] Additionally, microbiological results were assessed at the TOC visit.

[0128] At early time of cure (TOC) (predefined at day 8), 20% of patients in the placebo group were cured compared with 50% of patients in the high-dose CAL02 group. At TOC (predefined between days 15 and 22), all surviving patients were cured. The median time to cure was 10 days in the placebo group and 8 days in the high-dose CAL02 group (Table 6).

[0129] JPEG2025118655000008.jpg74170

[0130] A faster clinical recovery was observed in the CAL02 group compared to the placebo group: a 50% reduction in the SOFA score was already achieved in the CAL02 group after 5 days (Figure 1A). The APACHE II score also decreased more rapidly with CAL02 (high and low doses) compared to placebo (Figure 1B).

[0131] Pharmacodynamic biomarkers (CRP, PCT, IL-6) also returned to normal more rapidly with CAL02 (high and low doses) compared with placebo (Figure 2).

[0132] The length of stay in the ICU was significantly reduced from 12 days in the placebo group to 5 days in the CAL02 high-dose group (p<0.05), and the length of hospital stay was also reduced from 21 days in the placebo group to 12 and 12.5 days in the CAL02 high-dose and low-dose groups, respectively (Figure 3).

[0133] The impact of CAL02 treatment was also assessed as standalone or composite endpoints on: Rate of pneumonia recurrence and / or reinfection Chest X-ray changes with resolution of infiltrates and pneumonia Arterial blood gas generation by PaO2 / FiO2 ratio The need and duration of invasive or non-invasive assisted ventilation Duration of antibiotic therapy · Use of concomitant medications and rescue antibiotics. Other pharmacodynamic markers such as pancreatic stone protein (PSP) Vital signs (including heart rate, systolic and diastolic blood pressure, and core temperature) Hematology variables (including red blood cell count, hemoglobin and hematocrit, and white blood cell count including neutrophils, lymphocytes, monocytes, eosinophils, basophils, and platelets) Clinical chemistry variables (including sodium, potassium, CRP, total bilirubin, serum creatinine, glucose, BUN, calcium, inorganic phosphorus, chloride, albumin, alkaline phosphatase, total protein, lactate dehydrogenase (LDH), cholesterol, alanine aminotransferase (ALT=SGPT), aspartate aminotransferase (AST=SGOT), triglycerides, and uric acid) Urinalysis (including glucose, protein, bilirubin, urobilinogen, and nitrites) Physical examination (including general appearance, skin, respiratory, circulatory, abdominal, and nervous system) Microbiological results of CAP (respiratory, blood, pleural fluid samples) to determine eradication or presumed eradication, or persistence or presumed persistence, or superinfection or recurrence or new infection or colonization .

[0134] Conclusion: This is the first clinical trial to evaluate the potential of CAL02, a first-in-class, broad-spectrum antitoxin. The primary safety endpoint was met. Despite the fact that patients treated with CAL02 were more severely affected than those in the placebo group, patients treated with CAL02 showed more rapid clinical improvement. The efficacy trend was consistent across a range of clinical and biological parameters, demonstrating both faster resolution of organ dysfunction and more rapid reduction in pharmacodynamic biomarkers. These observations are consistent with CAL02's mechanism of action, which is intended to protect against toxin-mediated organ damage and inflammation.

[0135] Example 2 This randomized, multicenter, double-blind, placebo-controlled study will evaluate intravenous CAL02 as an adjunct to standard antibiotic therapy in patients admitted to the emergency room (ER), intermediate care unit, or intensive care unit (ICU) with proven or suspected infections requiring (i) intravenous antibiotics and (ii) management in an ICU or intermediate care unit. Patients will receive CAL02 or placebo in addition to standard antibiotic therapy. Infections include community-acquired pneumonia, hospital-acquired pneumonia, and ventilator-associated pneumonia, as well as intra-abdominal infections, skin and soft tissue infections, urinary tract infections in patients over 70 years of age, or bacteremia.

Claims

1. 1. A composition for use in the adjunctive treatment of pneumonia in a human patient, comprising, preferably consisting of, a mixture of empty liposomes, said mixture of empty liposomes comprising: (a) a first empty liposome comprising cholesterol, the first empty liposome having an amount of cholesterol of at least 30% (weight per weight); (b) a second empty liposome comprising sphingomyelin, preferably comprising only said sphingomyelin as lipid component; Preferably, said pneumonia is selected from Community Acquired Pneumonia (CAP), Hospitalization Acquired Pneumonia (HAP) and Ventilator Associated Pneumonia (VAP) in human patients.

2. 2. The composition for use according to claim 1, wherein the amount of cholesterol in the empty liposome (a) is between 45% and 55% (weight per weight) and the second empty liposome (b) consists of sphingomyelin.

3. 3. The composition for use according to any one of claims 1 to 2, wherein the first empty liposome (a) consists of cholesterol and sphingomyelin, the amount of cholesterol in the empty liposome (a) is about 50% (weight per weight), and the mixture of empty liposomes comprises at least 40%, preferably at least 45% (weight per weight) of the first (a) and second (b) empty liposomes.

4. 4. The composition for use according to claim 1, wherein the first empty liposome (a) consists of a 1:1 (weight per weight - w / w) mixture of the first empty liposome and the second liposome, the first empty liposome being composed of cholesterol and sphingomyelin in a 1:1 weight ratio (1:1 w / w, 35:65 molar ratio), the second empty liposome being composed of sphingomyelin alone, the first empty liposome (a) comprising only the cholesterol and the sphingomyelin as lipid components, and the second empty liposome (b) comprising only the sphingomyelin as lipid component.

5. The composition for use according to any one of claims 1 to 4, wherein said pneumonia is community-acquired pneumonia (CAP).

6. 6. The composition for use according to any one of claims 1 to 5, wherein said pneumonia is severe pneumonia, preferably said severe pneumonia is selected from severe community-acquired pneumonia (sCAP) and severe community-acquired pneumococcal pneumonia (sCAPP).

7. The pneumonia is preferably severe pneumonia, and the pneumonia is preferably caused by Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecium, Legionella pneumophila, Haemophilus influenzae, Klebsiella pneumoniae, or any of the following:

7. The composition for use according to any one of claims 1 to 6, wherein the bacterial infection is caused by Acinetobacter pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Serratia marcescens, Stenotrophomonas maltophilia, Moraxella catarrhalis, or Mycobacterium tuberculosis.

8. The composition for use according to any one of claims 1 to 7, wherein the pneumonia is caused by Streptococcus pneumoniae.

9. The composition for use according to any one of claims 1 to 8, wherein the treatment is adjunctive to antibiotic therapy.

10. 10. The composition for use according to claim 9, wherein the antibiotic therapy is intravenous (IV) or oral antibiotic therapy.

11. 11. The composition for use according to any one of claims 1 to 10, wherein the composition is administered in at least two doses, a first dose and a second dose, and the interval between the first and second dose is between 6 and 96 hours, preferably between 12 and 72 hours, more preferably between 24 and 48 hours, and even more preferably 24 or 48 hours.

12. The composition for use according to any one of claims 1 to 11, wherein the composition is administered in the form of a solution for intravenous administration.

13. 13. The composition for use according to any one of claims 1 to 12, wherein the composition is administered in at least two doses, the first dose being administered within 24 hours after the start of the antibiotic therapy, preferably IV antibiotic therapy.

14. 14. The composition for use according to any one of claims 1 to 13, wherein said pneumonia, preferably said severe pneumonia, more preferably said sCAP, requires hospitalization, preferably hospitalization of said patient in an intensive care unit (ICU), and wherein said treatment is adjunctive to antibiotic therapy, and wherein said adjunctive treatment reduces the duration of said hospitalization compared to hospitalization in the absence of such treatment.

15. 15. The composition for use according to claim 14, wherein the reduction in the length of hospital stay due to the adjunctive treatment is at least 1 day, preferably 2 days, more preferably 3 days, even more preferably 4 days, even more preferably 5 days, even more preferably 6 days, and even more preferably 7 days.

16. 16. The composition for use according to any one of claims 1 to 15, wherein the pneumonia, preferably the severe pneumonia, more preferably the sCAP, is cured in a shorter time compared to when such an adjunctive treatment is not performed, preferably when such a treatment is not performed adjunctive to the antibiotic therapy.

17. 17. The composition for use according to claim 16, wherein said faster cure is represented by a normalization of organ function that is at least 1 day shorter, preferably 2 days shorter or even more preferably at least 3 days shorter compared to when no such adjunctive treatment is given, preferably when no such treatment adjunctive to said antibiotic therapy is given.

18. 18. The composition for use according to any one of claims 16 to 17, wherein said faster healing is manifested by a normalization of blood inflammatory markers, preferably CRP and / or PCT and / or IL-6, that is at least 1 day shorter, preferably 2 days shorter or even more preferably at least 3 days shorter compared to the case in which no such adjunctive treatment is performed, preferably no such treatment adjunctive to said antibiotic therapy.

19. 19. The composition for use according to any one of claims 16 to 18, wherein said faster healing is represented by a reduction in SOFA score of at least 1 day shorter, preferably 2 days shorter or even more preferably at least 3 days shorter compared to the case in which no such adjunctive treatment is given, preferably no such treatment adjunctive to said antibiotic therapy.

20. 20. The composition for use according to any one of claims 16 to 19, wherein said faster healing is reflected by a reduction in APACHE II score of at least 1 day shorter, preferably 2 days shorter or even more preferably at least 3 days shorter compared to the absence of such adjunctive treatment, preferably the absence of such treatment adjunctive to said antibiotic therapy.

21. 21. The composition for use according to any one of claims 1 to 20, wherein said treatment of said pneumonia, preferably said severe pneumonia, more preferably said sCAP, even more preferably said sCAPP reduces the SOFA score of said patient by at least 40% 8 days after administration of the first dose of said composition, preferably 7 days after administration of the first dose of said composition, more preferably 6 days after administration of the first dose of said composition, even more preferably 5 days after administration of the first dose of said composition.

22. 22. The composition for use according to any one of claims 1 to 21, wherein said treatment of said pneumonia, preferably said severe pneumonia, more preferably said sCAP, even more preferably said sCAPP reduces the APACHE II score of said patient by at least 20% 8 days after the first administration of said composition, preferably 6 days after the first administration of said composition, more preferably 5 days after the first administration of said composition, even more preferably 4 days after the first administration of said composition.

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